Table of Content

21 March 2019, Volume 40 Issue 2

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Cancer nutrition in China: development and challenges

Shi Hanping, Jia Pingping

2019, 40(2):  159-162.  doi:10.3969/j.issn.1006-7795.2019.02.001

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In China, the incidence of cancer malnutrition is high and it leads to poor clinical outcome, increased complications, extension of hospital stay and shortened survival time. Misunderstanding on cancer nutrition therapy and non-standard treatment restricts the development of cancer nutrition. In recent years, the development of cancer nutrition in China has attracted more attention and made a great progress:①exploring the molecular mechanisms and the individual differences of nutritional metabolism based on the characteristics of tumor metabolism, which provide the basis for individualized nutritional therapy; ②conforming the cancer nutritional status in China:the high incidence of severe malnutrition, lack of nutrition knowledge in patients and medical personnel, improper practice,which is helpful for government policies making and the development of disciplines and clinical treatment;③established the disciplines of cancer nutrition, the diagnosis and treatment system of cancer nutrition in China. Proposed three level diagnosis, four dimensional analysis and five ladder therapy of malnutrition;④Promoting the awareness of cancer nutrition, standardizing diagnosis and treatment,which promote the application and popularity of cancer nutrition care. However, further work is needed to revise the awareness and remedy inappropriate management of cancer nutrition in China. Ultimately it bring nutrition care to the first-line therapy.

Study of allogeneic stem cell transplantation for pediatric acute myeloid leukemia

Jia Chenguang, Zhou Xuan, Wang Bin, Zhu Guanghua, Qin Maoquan

2019, 40(2):  163-168.  doi:10.3969/j.issn.1006-7795.2019.02.002

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Objective To analysis the effectiveness of allogeneic hematopoietic stem cell transplantation (allo-HSCT) for treatment of children with acute myeloid leukemia (AML), and to identify the possible prognostic factors for treatment. Methods We retrospectively reviewed outcomes of children with AML who underwent allo-HSCT at Hematology Oncology Center of Beijing Children's Hospital, Capital Medical University between January of 2006 and January of 2016. The overall survival (OS) were estimated by Kaplan-Meier method, Log-rank test was used to compare the different groups and prognostic factors were evaluated by Cox regression. Results A total of 46 patients were involved, with 31 patients survived, and 15 patients died, 8 patients relapsed, with an average follow-up time of 27 months. 5 years OS was (61.1±8.7)%, median survival time was 77.3 months, and 5 years cumulative relapse rate was(16.0±6.0)%. The results of Log-rank test indicated no significant difference (P>0.05) with different protocol, remission state, remission times, graft versus-host disease (GVHD) and donors. The Cox regression suggested myeloid sarcoma and the relapse are the prognostic factors (P<0.05). Conclusion Allogeneic hematopoietic stem cell transplantation is an effective treatment for AML, especially, refractory/relapsed AML. With the modified conditioning regimen and supportive treatment, the survival rate is increasing year by year. Myeloid sarcoma and relapse are important adverse prognostic factors of HSCT treatment for AML.

Clinical features and outcome evaluation for children with hepatoblastoma

Wang Xisi, Yang Wei, Huang Cheng, Zhao Wen, Zhao Qian, Duan Chao, Su Yan, Jin Mei, Zhang Dawei, Qin Hong, Wang Huanmin, Ma Xiaoli

2019, 40(2):  169-173.  doi:10.3969/j.issn.1006-7795.2019.02.003

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Objective To summarize the clinical characteristics and treatment effect of hepatoblastoma (HB) in Beijing Children's Hospital in the past ten years, and further to provide the basis for reasonable diagnosis and treatment strategy to reduce the side effects and improve the event-free survival. Methods The patients who received multidisciplinary medical therapy, including surgery, radiotherapy and chemotherapy, and followed up regularly were enrolled into the group. The chemotherapy includes regimen A (vincristine + cisplatin + fluorouracil) and regimen B (cisplatin + doxorubicin). We recorded the age, sex, course of disease, initial serum alpha fetal protein(AFP), clinical stage, pathological type, treatment and treatment effect of all the patients. The cut-off data of follow-up was November 30, 2018. Results In total 51 cases admitted in our hospital, 42 cases were admitted into the research group (six cases were treated in other hospitals, three cases died because of the tumor rupture before treatment). According to the cases of medical recorder in the group, the median age was 19 months (2-102 months); 1 case in stage Ⅱ, 34 cases in stage Ⅲ, and 7 cases in stageⅣ. The value of AFP was in the range of 31.6-2 200 000 μg/L(median data was 192 274.5 μg/L).The median follow-up was 36 months (6-115 months) in 39 cases, and the 5-year event-free survival (EFS) was 74.72%. After the chemotherapy, the whole blood cells decreased insignificantly. Neither severe infection nor cardio toxicity and hearing impairment occurred during chemotherapy. Conclusion Comprehensive treatment and management of surgery combined with chemotherapy is the main treatment for children with HB. The dosage of doxorubicin and cisplatin in the current regimen is tolerable, with fewer side effects, and the 5-year EFS is better. Long-term follow-up of these children is needed in the future.

A evaluation of the early efficacy and safety of generic imatinib in treating children of chronic myeloid leukemia in chronic phase

Zhang Liqiang, Chen Zhenping, Zheng Jie, Ma Jie, Wu Runhui

2019, 40(2):  174-178.  doi:10.3969/j.issn.1006-7795.2019.02.004

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Objective To evaluate the hematological, cytogenetic, molecular responses and safety of generic imatinib(Xinwei) in newly diagnosed patients with children of chronic phase of chronic myeloid leukemia (CP-CML). Methods Thirty-five CP-CML children patients diagnosed in Beijing Children's Hospital, Capital Medical University received oral imatinib 260-340 mg/m²,with domestic generic imatinib (Xinwei) and imported original imatinib (Gleevec), who were collected from Jan 2014 to Jan 2018 and followed up to May 2018. Compared with imported original imatinib (Gleevec) among which 20 cases were treacted with the domestic genenc imatinib (Xinwei) and 15 cases were treated with the imported original imatinib (Gleevec), hematological responses, cytogenetic examinations, BCR-ABL transcript levels, and the safety with domestic generic imatinib treatment were monitored after 3,6, and 12 month treatment. Results A total of 35 children CP-CML patients (18 males and 17 females with a median age of 10 years) were collected among which 20 cases were treated with the domestic generic imatinib (Xinwei), and 15 cases were treated with the imported original imatinib (Gleevec). At 3-month, the domestic generic imatinib group 75% (15/20) patients achieved the complete hematologic responses (CHR), and patients with minor cytogenetic response (MCyR) or BCR-ABL ≤ 10% were 80%(16/20) and 45%(9/20), respectively. At 6-month, 94%(15/16) patients achieved CHR, patients with complete cytogenetic response(CCyR) and BCR-ABL ≤ 1% was 62.5%(10/16) and 43.8%(7/16). At 12-month, 92.3%(12/13) CCyR and 77%(10/13)BCR-ABL ≤ 0.1%. There was no significant difference between two group at all stages (P>0.05). The hematologic toxicity occurred in 15%(3/20),with no grade Ⅲ hematologic toxicity. The non-hematologic toxicities included pain 35%(7/20),gastrointestinal reaction 25%(5/20), edema 15%(3/20), fever 10%(2/20),fatigue 5%(1/20), and rash 5%(1/20). Compared with imported original imatinib, CCyR was lower in generic imatinib group at 3-month (P=0.021). There was no other significant difference (P>0.05). The adverse reactions were easy to control and there were no drug-related deaths.Conclusion Generic imatinib has a good efficacy in the hematological, cytogenetic molecular reponses with CP-CMP patients in newly diagnosed, without serious adverse reactions. There is no significant difference in efficacy and safety between generic and imported original imatinib in our study.

DCLK1 alternative variants enhance proliferation of pancreatic cancer cells by activating JNK pathway

Zhang Yuanyuan, Ge Yang, An Guangyu

2019, 40(2):  179-185.  doi:10.3969/j.issn.1006-7795.2019.02.005

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Objective To investigate the effects of long-and short-isoform of doublecortin-like kinase 1 (DCLK1) on the proliferation of human pancreatic cancer and further to explore the molecular mechanism. Methods The control (PCMV6-AC-GFP), DCLK1 isoform 1 and DCLK1 isoform 4 eukaryotic expression plasmids were transfected into pancreatic cancer PANC-1 cells. G418 screening method was used to construct stable cell lines overexpressing long-and short-isoform of DCLK1. Expression of long-and short-isoform of DCLK1 were determined by quantitative real time polymerase chain reaction(qRT-PCR) and Western blotting. The effect of long-and short-isoform of DCLK1 on the proliferation of PANC-1 cells were detected with real-time cell analysis (RTCA) technology. The effect of DCLK1 on mitogen-activated protein kinase (MAPK) pathway was detected with Western blotting. DCLK1 stabilizing cells were treated with specific c-Jun N-terminal kinase (JNK) pathway inhibitor SP600125 to detect the effect of JNK pathway inhibition on the proliferation of pancreatic cancer cells. Results Overexpression of long-and short-isoform of DCLK1 remarkably promoted the proliferation of pancreatic cancer cells (P<0.05), increased the phosphorylation of JNK in MAPK pathway and the expression of target molecules CMYC, CD44 and CyclinD1 in JNK pathway (P<0.05). No significant influence on the phosphorylation of ERK and p38 were detected (P>0.05). SP600125 inhibited the phosphorylation of JNK and markedly decreased the activation of JNK pathway and the proliferation of pancreatic cancer cells by DCLK1 (P<0.05). Conclusion Both long-and short-isoform of DCLK1 promote the proliferation of pancreatic cancer cells via activating JNK pathway.

Effects of β-hydroxybutyrate on the proliferation and glycolysis in glioma cells

Yu Chunna, Jiang Bo, Li Wenbin, Chen Feng

2019, 40(2):  186-190.  doi:10.3969/j.issn.1006-7795.2019.02.006

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Objective To investigate the effects of β-hydroxybutyrate (β-HB), one of the ketogenic diet (KD) metabolites, on the proliferation and glycolysis of glioma cell lines to reveal the mechanisms of KD in the treatment of glioma. Methods After treated with different concentrations of β-HB, the proliferation of glioma cell lines U87 and LN229 was detected with CCK8 assay and glycolysis ability of cells was measured with glycolysis stress test. Further, the transcription factor c-myc expression was detected with Western blotting and real-time quantitative polymerase chain reaction (RT-qPCR). Results β-HB can inhibit the proliferation of glioma cells in a dose-dependent manner. Compared with the untreated group, the glycolysis level and maximum glycolytic capacity was decreased and the expression of c-myc was down-regulated in the experimental groups. Conclusion β-HB can affect the glycolytic capacity of glioma cell lines U87 and LN229 through c-myc, and thus inhibit cell growth. It is suggested that KD can be used as an efficacious therapy agent for glioma.

Serum lipid and apolipoprotein levels were associated to the Ki-67 and molecular classification of breast cancer

Chen Nina, Fan Yichang, Shang Kun, Yang Jing, Cao Bangwei

2019, 40(2):  191-194.  doi:10.3969/j.issn.1006-7795.2019.02.007

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Objective To investigate the relationship between serum lipid and molecular types, Ki-67 level of breast cancer. Methods Total of 143 breast cancer patients who underwent the surgery from January 2012 to May 2017 were enrolled. The level of high density lipoprotein-cholesterol (HDL-C), low density lipoprotein-cholesterol (LDL-C), cholesterol (TC), apolipoprotein B (ApoB), apolipoprotein A (ApoA),and triglyceride (TG) were measured. The relationship between the above factors and the different molecular types of breast cancer was analyzed and discussed. Results The level of HDL-C, LDL-C, TC, ApoB and TG were significantly associated with different breast cancer subtypes (P<0.05). No significant difference of ApoA levels was observed in the different breast cancer subtypes (P>0.05). The levels of LDL-C, TC and ApoB were positively correlated with Ki-67 levels, while the levels of HDL-C was negatively correlated with Ki-67 level. However, The levels of ApoB and TG showed no relation with Ki-67 expression level. Conclusion Lipid metabolism is different in different breast cancer molecular subtypes. The serum lipid level is related to the expression level of Ki-67. High levels of TC and LDL-C and low level of HDL-C suggest poor prognosis in breast cancer.

Early diagnosis of pancreatic cancer in a nude mice model on synchrotron radiation X-ray phase contrast imaging

Wang Jia, Le Kai, Guo Yifan, Guo Xu, Zhang Tao, Ren Yu, Wang Ruibin, Xu Guangzhong, Zheng Hailiang, Feng Shuli, Zhu Bin

2019, 40(2):  195-200.  doi:10.3969/j.issn.1006-7795.2019.02.008

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Objective To explore the diagnostic value of synchrotron radiation X-ray phase contrast imaging compared with magnetic resonance imaging (MRI) in early diagnosis of pancreatic cancer with an orthotopic xenograft nude mice model of pancreatic cancer. Methods Thirty-two female nude mice were divided into 2 groups of 4 subgroups randomly, subgroups were labeled A1, A2, A3, A4 (n=6) and B1, B2, B3, B4 (n=2) as the different intervals at 1, 2, 3, and 4 weeks postoperatively, respectively. 10⁸ cells/mL×20μL PANC-1 cell suspension was injected into pancreatic capsula of group A but 20μL sterile saline was injected in the group B. MRI scans were performed at 1, 2, 3, and 4 weeks postoperatively and the specimens were excised from nude mice and fixed in 10% formalin immediately. Synchrotron radiation X-ray phase contrast imaging was performed followed by histopathological examination. Results ①Low grade adenocarcinoma was confirmed with pathology, and the average rate of tumorigenesis in group A was 73% (16/22), and the rate was the highest after 4 weeks, which was 100%. With the time passing, the rate of tumorigenesis increased with the linear trend test (χ²=4.718, P=0.030). ②The image of tumor with a diameter of 0.5 mm in pancreas was observed with synchrotron radiation phase contrast imaging. The sensitivity and specificity of the early (two weeks) reached 100%, significantly higher than that of magnetic resonance imaging (P=0.029). Conclusion PANC-1 cells injected into pancreatic capsula was a reliable method for establishment of pancreatic cancer model. Compared with MRI, synchrotron radiation contrast imaging could detect tiny lesions of pancreatic cancer, which may provide a new method for early diagnosis of pancreatic cancer.

Tumor necrosis factor-α and interferon-γ in cerebrospinal fluid of children with medulloblastoma

Zhang Jin, Ren Siqi, Li Shuting, Li Miao, Gong Xiaojun, Sun Yanling, Wu Wanshui, Sun Liming, Du Shuxu

2019, 40(2):  201-204.  doi:10.3969/j.issn.1006-7795.2019.02.009

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Objective To evaluate the diagnostic value of tumor necrosis factor-α(TNF-α) and interferon-γ(IFN-γ) in cerebrospinal fluid (CSF) for the prognosis of medulloblastoma. Methods From November 2016 to June 2018, 167 children with medulloblastoma from Beijing Shijitan Hospital, Capital Medical University were enrolled in this study. They were 113 boys and 54 girls, with the median age 7.6 years old (ranged from 14 months to 17 years old). They were divided into 3 groups:non-metastasis group, metastasis group, and recurrent group. CSF specimens were collected before chemotherapy. Enzyme linked immunosorbent assay (ELISA) was used to detect TNF-α and IFN-γ in CSF. The diagnostic value for prognosis was analyzed with receiver operating characteristic (ROC) curve according to area under the ROC curve (AUC), cutoff value, sensitivity (SEN), specificity (SPE), and Youden index (YI). Results The levels of TNF-α and IFN-γ were significantly higher in the non-metastasis group than that in the metastasis and recurrent group (P<0.05). There was significant difference for IFN-γ between the metastasis and recurrent group (P<0.05), and no significant difference for TNF-α(P>0.05). The ROC curves showed that the AUC of TNF-α was 0.958, and SEN of 90.9%, SPE of 87.1%, YI of 0.78 when the cutoff value was 12.73 ng/L. The AUC of IFN-γ was 0.809, and SEN of 78.8%, SPE of 80.2%, YI of 0.59 when the cutoff value was 136.53 ng/L. Conclusion TNF-α and IFN-γ play a role to fight against tumor cells in medulloblastoma, and can be used as a valuable evaluation marker of prognosis. The diagnostic value of TNF-α is higher than that of IFN-γ.

Current status of research concerning neuroinflammation schizophrenia

Ma Chao, Jia Hongxiao

2019, 40(2):  205-208.  doi:10.3969/j.issn.1006-7795.2019.02.010

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Schizophrenia is a common mental disorder, the mechanism of which remains unknown at present. Many hypotheses have been put forward. As early as the 1980s, three were the clinical studies showing that schizophrenia activated the inflammatory response and the immune system disorders in the peripheral nervous system and the brain. In recent years, psychiatric drugs and anti-inflammatory drugs are used in combination to improve the symptoms in schizophrenic patients, suggesting that nerve inflammation played a role in the onset of schizophrenia. The neuroinflammation effects on schizophrenia are reviewed in this article.

Effects of Yishenpingganfang, an empirical prescription on social interaction behavior in mice exposed to cuprizone and its possible action target

Wu Yan, Sun Zuoli, Ma Chao, Zhu Hong, Yin Dongqing, Jia Hongxiao

2019, 40(2):  209-213.  doi:10.3969/j.issn.1006-7795.2019.02.011

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Objective To explore the effect and potential target of Yishenpingganfang (YSPGF) in schizophrenia treatment. Methods Totally 30 Young adult male C57BL/6 mice (6 weeks old) were randomly divided into three groups:normal control, cuprizone (CPZ) model, and YSPGF group. This study lasts for 6 weeks. Then, social interaction tests and brain tissue collections were carried out 24h later after 2-week period of drug admission. Social interaction was used to evaluate schizophrenia-like symptoms in mice exposed to cuprizone and partly treated by YSPGF. The mature oligodendrocyte/myelin deficiency and immature oligodendrocyte changes were observed with histological staining. Results The body weight in the model group were obviously decreased in week 3 compared with the control group (P<0.05). Social interaction included two trials:In trial 1, control mice spent more time interacting with the stranger mouse than with the empty cage and familiar one (P<0.001) as well as YSPGF group (P<0.01).In trial 2, control mice preferred to be near the new stranger mouse than the familiar one (P<0.001). In contrast, CPZ mice showed no preference between the empty cage and the stranger mouse in both trials 1 and 2.The oligodendrocyte/myelin deficiency existed in callosum through using mature oligodendrocyte maker myelin basic protein (MBP). Furthermore, we used immature oligodendrocyte maker platelet-derived growth factor receptor-alpha (PDGFR-α) and found that in model group mice these immature oligodendrocyte increased after the demyelination stage. Conclusion YSPGF could improve CPZ-induced psychosis-like behaviors and reduce oligodendrocyte/myelin deficiency.

Study on quality and satisfaction of life and the social function in patients with depersonalization disorder

Wang Rui, Song Nan, Zhu Hong, Jia Hongxiao

2019, 40(2):  214-219.  doi:10.3969/j.issn.1006-7795.2019.02.012

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Objective To Explore the quality and satisfaction of the life and social function in patients with depersonalization disorder(DPD). Methods The total thirty-eight patients with depersonalization disorder who were diagnosed with the diagnostic criteria of the Diagnostic and Statistical Manual of Mental Disorders, Fifth Edition (DSM-V), and the Cambridge Depersonalization Scale(CDS) of the patients scored ≥ 70. Forty normal persons were set as the control. All the patients and controls were evaluated with the Global Assessment Function (GAF), the Short-Form Version of the Quality of Life Enjoyment and Satisfaction Questionnaire(Q-LES-Q-SF) and the World Health Organization Quality of Life Brief questionnaire (WHOQOL-BREF) to assessed quality and satisfaction of the life and social function. Results Compared to the control group, the total scores of GAF and WHOQOL-BREF in the patients were significantly lower on the quality of the life(PHYS), psychological field (PSYCH), social relations field (SOCIL), environmental field (ENVIR), and the total scores of Q-LES-Q-SF. The score on GAF in DPD was 61.21±5.99, vs 85.88±2.81 in normal group. The score on PHYS in DPD was 11.17±1.87, vs 12.96±1.96 in normal group. The score on PSYCH in DPD was 10.67±1.78, vs 13.33±2.32 in normal group. The score on SOCIL in DPD was 10.53±2.90, vs 14.50±2.64 in normal group. The score on ENVIR in DPD was 12.00±2.61, vs.13.45±2.18 in normal group. The total score on Q-LES-Q-SF in DPD was 39.26±7.93, vs 50.88±8.38 in normal group. The total score of Cambridge Depersonalization Scale was correlated with psychological field, social relationship field, environment field, total quality of life and satisfaction (P<0.05). Compare to simple DPD, the total scores of GAF were lower in DPD with anxiety. The score on GAF in simple DPD was 57.67±5.69 vs 62.85±5.48 in DPD with anxiety(P<0.05). Conclusion The quality and satisfaction of life and social function of DPD was significantly worse than that of normal controls, regarding not only in the physiological aspect but also to the patient's emotional experience, social function and mental health level.

Effect of Jinkui Shenqi Pill on serum nerve growth factor in depression patients with kidney deficiency and liver stagnation

Li Miao, Wang Qunsong, Yuan Guozhen

2019, 40(2):  220-225.  doi:10.3969/j.issn.1006-7795.2019.02.013

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Objective To investigate the effect of Jinkui Shenqi Pill on serum nerve growth factor (NGF) in depression patients with kidney deficiency and liver stagnation. Methods The patients were divided into the trail group (30 cases) and control group (30 cases) by random number table method. The trail group were treated with Jinkui Shenqi Pill combined with escitalopram oxalate, one dose a day given in the morning and evening. The control group were treated with escitalopram oxalate at a dose 20 mg/d alone. The treatment course was 8 weeks. The Montgomery-Asberg-Rating Scale (MADRS) and traditional Chinese medicine (TCM) symptoms scale were measured before and at the end of weeks 2, 4 and 8, and the serum NGF levels were also recorded. Results There was no significant difference in MADRS and TCM symptoms scores before treatment between the two groups (P>0.05). The evaluation interval (2, 4, and 8 weeks) was taken as the intra-group factor and the group (trail group and control group) as the inter-group factor. By repeated measurement of variance analysis, the intra-group factor (MADRS) independent variable effect was significant[F_(2,58)=301.367,P=0.000], and the interaction between intra-group and inter-group factors was significant[F_(2,58)=4.022,P=0.008] while inter-group factor independent variable effect was significant[F_(2,58)=9.658,P=0.003];the intra-group factor (TCM symptoms) independent variable effect was significant[F_(2,58)=355.130,P=0.000], and the interaction between intra-group and inter-group factors was not significant[F_(2,58)=0.970,P=0.374],and the interaction between intra-group and inter-group factors was significant[F_(2,58)=5.336,P=0.024].It suggested that the therapeutic efficiency of the trail group is better than that of the control group. The levels of NGF before treatment in both groups were equivalent in general,with no significant difference (P>0.05).After the treatment, the NGF level in the trail group was high than that in the control group with significant difference (P<0.05). Pearson correlation analysis showed that the difference of NGF levels before and after treatment in the trail group was positively correlated with the difference of kidney deficiency score (r=0.481,P=0.007) and the difference of liver stagnation score (r=0.370,P=0.044).Conclusion Jinkui Shenqi Pills can effectively improve the symptoms of depression patients with kidney deficiency and liver depression.Compared to the patients using escitalopram oxalate alone, the serum NGF concentration in the combinatorial group increased significantly.

Optimization of induction and differentiation of 3T3-L1 preadipocyte

Zhao Lei, Zheng Meili, Yang Mei, Zhong Jiuchang, Yang Xinchun

2019, 40(2):  226-231.  doi:10.3969/j.issn.1006-7795.2019.02.014

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Objective To optimize the methods of the induction and differentiation of 3T3-L1 preadipocytes. Methods Based on the "cocktail" method, 3T3-L1 preadipocytes were divided into four groups according to the differentiation medium A:(1) Control group; (2) Ros:rosiglitazone group; (3) Indo:rosiglitazone group; (4) Ros+Indo:rosiglitazone+indometacin combination group. During the differentiation, cell morphology was observed through microscope. Oil red O was stained and quantified. The intracellular and extracellular triglyceride (TG) content was measured via TG GPO-PAP enzymatic kit. Results The differentiation rates were higher in group "Ros", "Indo" and "Ros+Indo", with earlier adipogenesis and more lipid droplets, which was further demonstrated in oil red O staining. In the evaluation of one simple lipid droplet with three parameters:mean diameter, mean area and max diameter, the droplet in group "Ros", "Indo" and "Ros+Indo" were much bigger than that in control group, while there was no statistically significance between control and "Indo" group. Regarding TG content, there was no significant difference among four groups in extracellular TG, but intracellular TG was higher in "Ros+Indo" group than that in control group (0.77±0.07 mmol/g vs 0.19±0.02 mmol/g, P<0.05). Conclusion Based on the classical "cocktail" method, PPAR-γ agonist rosiglitazone significantly improves the differentiation rate of 3T3-L1 preadipocytes.

Establishment of a serum-free primary culture method for enteric glial cell of myenteric plexus

Quan Zhusheng, Zhang Xiaoli, Zhu Jinxia

2019, 40(2):  232-236.  doi:10.3969/j.issn.1006-7795.2019.02.015

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Objective Enteric glial cell (EGC) plays an important role in regulation of many physiological and pathological process in the intestine, in order to further study the function of EGC and exclude the influence of neurohumoral factors at the vivo and vitro level, it is necessary to obtain a simple and efficient method for isolation and primary culture of EGC. Methods Colonic myenteric plexus were digested and separated. The fibroblasts were removed by alternating culture with serum-containing and serum-free medium and then subcultured routinely. The purity and activity of primary EGC were detected by immunofluorescence and intracellular Ca²⁺ imaging. Results The number of EGC was about 6.8×10⁵ on each experiment before passage. After serum-free culture, fibroblasts decreased significantly; the percentage of immunoreactivities of glial fibrillary acidic protein(GFAP), S100β and both GFAP and S100β was 98.44%±1.07%, 93.61%±3.16% and 93.09%±2.99%, respectively.About 96% EGC manifested intracellular calcium transient when treated with ATP. Conclusion Combined with the serum-free culture,fibroblasts were removed and primary myenteric EGC with higher purity and activity could be obtained.

Progesterone receptor membrane component 1 increases hormone induced proliferation of breast cancer cells——E2 vs sequential combination vs continuous combination

Li Xue, Ruan Xiangyan, Gu Muqing, Cai Guiju, Zhao Yue, Alfred O. Meuck

2019, 40(2):  237-243.  doi:10.3969/j.issn.1006-7795.2019.02.016

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Objective To investigate the effect of progesterone receptor membrane component 1 (PGRMC1) on the risk of hormone induced breast cancer, and compare the effects of different treatment regimens:estradiol (E2)/sequential combination/continuous combination(E2/DRSP、E2/NET) on breast cancer proliferation. Methods After stabilized transfection by lipofection, T47D cells transfected with vector(T47D-HA-vector) or PGRMC1(T47D-HA-PGRMC1) were stimulated with E2 alone (0.1, 0.01, 0.001 nmol/L), or sequential/continuous combination. CCK-2 method was performed to measure cell proliferation. Results When treated with E2 (0.001, 0.01, 0.1 nmol/L) alone, T47D-HA-vector cells did not show significant proliferation (P>0.05). At concentration of 0.1 nmol/L, T47D-HA-PGRMC1 cell reached a significant increase in cell proliferation compared with the control group (56%, P<0.05) and compared with T47D-HA-vector group (36%, P<0.05); when treated with sequential combination, T47D-HA-PGRMC1 cell under E2/NET treatment (E2=0.001 nmol/L) showed a significant increase in cell proliferation compared with the control group (82%, P<0.05) and T47D-HA-vector group (63%, P<0.05). Under the same stimulation, when the concentration of E2 reached 0.1 nmol/L, T47D-HA-vector showed a significant increase compared with the control group (37%, P<0.05), and for the T47D-HA-PGRMC1 cell line, when under E2/drospirenone (DRSP) or E2/norethisterone (NET) stimulation (E2=0.1 nmol/L), a significant increase compared with the control group (105%, 170%, P<0.05) and T47D-HA-vector group (84%, 133%, P<0.05) was observed; when treated with the continuous combination, the T47D-HA-PGRMC1 cell line showed a significant increase in cell proliferation under E2/DRSP or E2/NET stimulation (E2=0.001 nmol/L) compared with the control group (77%, 158%, P<0.05) and T47D-HA-vector group(60%, 136%, P<0.05). The T47D-HA-vector cell line showed a significant increase under E2/NET stimulation (E2=0.1 nmol/L) compared with the control group (44%, P<0.05), and the T47D-HA-PGRMC1 cell line under E2/DRSP or E2/NET stimulation (E2=0.1 nmol/L) showed a significant increase in cell proliferation compared with the control group (129%, 174%, P<0.05) and T47D-HA-vector group (97%, 131%, P<0.05). Conclusion PGRMC1 can significantly promote the proliferation of breast cancer cells induced by sequential/continuous combination. Compared with the sequential combination, the continuous combination promoted a higher proliferation of T47D.

Blood plasma of patients with Parkinson's disease increases alpha-synuclein neurotoxicity and its mechanisms

Li Jie, Xie Xiujuan, Song Wei, Pan Weihua, Yang Weiwei, Yu Shun

2019, 40(2):  244-250.  doi:10.3969/j.issn.1006-7795.2019.02.017

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Objective To investigate the effect of plasma of Parkinson's disease (PD) on the cytotoxicity of α-synuclein (α-Syn) and its mechanism. Methods Recombinant human α-Syn was added to PD and normal subject plasma. Western blotting and ELISA method were used to identify α-Syn oligomers and phosphorylation. Add α-Syn monomer and oligomer at a final concentration of 5μM to primary neuronal medium with 30% PD plasma, normal subject plasma and fetal bovine plasma, after 14 days of incubation, MTT and flow cytometry were used to observe the cell death. Enzyme activity kit was used to detect the activity of protein phosphatase 2A (PP2A) enzyme in primary neurons of each group. Results Plasma in PD patients induced more α-Syn phosphorylation and oligomerization than normal subjects (NS) plasma. Compared to NS plasma, PD plasma added to primary neuronal cultures resulted in more cells dying in the presence of extracellular α-Syn, accompanied by decreased protein phosphatase 2A activity. Conclusion The plasma of PD patients may phosphorylate α-Syn by decreasing the activity of PP2A enzyme, leading to aggregation and further death of primary cultured neurons.

Bufalin induces necroptosis in human gastric cancer SGC-7901 cells

Gong Pengchao, Li Yanlan, Kong Cuicui, Tian Xin

2019, 40(2):  251-256.  doi:10.3969/j.issn.1006-7795.2019.02.018

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Objective To investigate the necroptosis mechanism induced by Bufalin in human gastric cancer SGC-7901 cells. Methods SGC-7901 cells were cultured in vitro. The cells in the control group were cultured in RPMI 1640 medium, while those in the experimental group were exposed to various concentrations of Bufalin(50,100,150,200 nmol/L). After treatment for 48 h, the cell survival rate was assessed by MTT assay. Cell ultrastructures were observed by transmission electron microscope. Morphological changes in the nuclei were observed by DAPI fluorescence staining. Cellular necroptosis was assayed by flow cytometry. Western blotting was used to analyze the expression of receptor interacting protein kinase 1 (RIP1) in the necroptosis pathway. Results MTT assay showed that Bufalin had a significant effect on the growth of SGC-7901 cells(P<0.05). Transmission electron microscopy can be used to observe characteristic changes in cell necrosis, such as disruption of cell membranes, production of intracellular vacuoles, and disintegration of organelles. DAPI staining showed that after treatment with 100 nmol/L for 48 h, there were no obvious changes in cell apoptosis, such as nuclear condensation, nuclear fragmentation and formation of apoptotic bodies. The cell necrosis rate was higher than control group(P<0.05). Bufalin promotes the expression of RIP1, a key protein of necroptosis pathway(P<0.05). Conclusion Bufalin efficiently induced cell death through necroptosis pathway in SGC-7901 cells.

Angiotensin Ⅱ type 1 receptor autoantibody-positive female rats showed hypertensive disorder after pregnancy

Bian Jingwei, Yin Xiaochen, Li Hao, Zhang Suli, Wang Meili, Liu Huirong

2019, 40(2):  257-262.  doi:10.3969/j.issn.1006-7795.2019.02.019

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Objective To investigate whether angiotensin Ⅱ type 1 receptor autoantibody (AT1-AA)-positive rats exhibited the phenomenon of gestational hypertension after pregnancy. Methods Clinical serum samples were collected to detect the presence of AT1-AA in non-pregnant healthy women by enzyme linked immunosorbent assay (ELISA). AT1-AA-positive rat model was established by active immunization. Tail-cuff method was used to measure the systolic blood pressure and the mesenteric artery ring was used to detect vascular function. Results AT1-AA was also present in some healthy women who were not pregnant. There was no significant difference in blood pressure between AT1-AA-positive rats and control rats before pregnancy, but the systolic pressure of AT1-AA-positive rats after pregnancy was significantly higher than that of control rats. Mesenteric artery ring showed that the vasoconstriction was increased (vehicle group:110%±5%, immunized group:130%±10%, P<0.05), and endothelium-dependent/non-endothelium-dependent vasodilation was decreased in AT1-AA-positive rats after pregnancy (vehicle group:90%±4%, immunized group:60%±7%/vehicle group:95%±1%, immunized group:75%±3%, P<0.05). Conclusion AT1-AA-positive rats after pregnancy showed the phenomenon of gestational hypertension, such as elevated blood pressure and impaired vascular function. These suggested that the presence of AT1-AA in healthy individuals was a risk factor for gestational hypertension.

Synthesis of novel mononuclear Ru(Ⅱ) complexes with 10,13-diaryl substituents on the dppz ligands

Li Minna

2019, 40(2):  263-268.  doi:10.3969/j.issn.1006-7795.2019.02.020

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Objective Four new ruthenium complexes with 1,10-phenanthroline ancillary ligand and 10, 13-position substituted[pyrido-(3,2-a:2',3'-c)phenazine,dppz] major ligands including thiophenyl,furanyl,[2-benzo(b)thienyl]-2,1,3-benzothiadiazole and[2-benzo(b)furanyl]-2,1,3-benzothiadiazole will be designed and synthesized. Methods Target complexes were made by condensation reaction of ruthenium complex with the appropriate 3,6-diaryl substituted benzene-1,2-diamine. The latter 3,6-diaryl substituted benzene-1,2-diamine was obtained by a Suzuki-coupling reaction, and subsequently by reduction with NaBH₄ under catalysis of CoCl₂. Results Benzothiadiazole intermediates and corresponding target complexes are well characterized by ¹H nuclear magnetic resonance (NMR), ¹³C NMR and TOF-Mass. UV-vis absorption spectra of all ruthenium complexes further verify the structures of the target complexes. Conclusion In this paper, we have synthesized four mononuclear ruthenium complexes with 1,10-phenanthroline ancillary ligand and 10, 13-diaryl including thiophenyl,furanyl,[2-benzo(b)thienyl]-2,1,3-benzothiadiazole and[2-benzo(b)furanyl]-2,1,3-benzothiadiazole, substituted dppz as major ligand through multiple steps. Both four 4,7-position substituted 2,1,3-benzothiadiazole intermediates and corresponding ruthenium complexes exhibit substantially similar absorption spectra despite the structural difference.

The influence of 27-hydroxycholesterolon cholesterol transport associated factor in co-cultured SH-SY5Y cells and C6 cells

Chen Si, Zhou Cui, Yu Huiyan, Tao Lingwei, Wang Yushan, Xiao Rong

2019, 40(2):  269-275.  doi:10.3969/j.issn.1006-7795.2019.02.021

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Objective To investigate the effect of 27-hydroxycholesterol (27-OHC) on cell lysosomal cholesterol transport associated factor under the conditions of co-culture of nerve cells and astrocytes. Methods SH-SY5Y cells and C6 cells were co-cultured with trans-well chamber. The experiment was divided into control group (serum-free medium), 27-OHC 5 μmol/L group (low dose group), 27-OHC 10 μmol/L group (middle dose group) and 27-OHC 20 μmol/L group (high dose group). The total cholesterol levels were detected by total cholesterol kit. The expression level of mRNA and proteins of lysosomal cholesterol transport associated proteins (including NPC1 and NPC2) were detected by real-time PCR and Western blotting. Results The total cholesterol levels of cellular protein from the C6 cells treated with 5, 10 and 20 μmol/L 27-OHC was significantly decreased compared with control (P<0.05). However, SH-SY5Y cells in 27-OHC treatment groups had higher total cholesterol levels (P<0.05). Results showed reduced levels of lysosome cholesterol associated genes and proteins (NPC1 and NPC2) in SH-SY5Y cells (P<0.05) and increased levels in C6 cells (P<0.05). Conclusion Our data indicate that 27-OHC causes neuronal and astrocyte lysosomal cholesterol transport disorders,and accumulate lysosomal cholesterol in nerve cells.

Analysis of influence factors of suboptimal health status: undergraduates of Weifang University in 2017

Sun Qi, Wang Hao, Liu Hongqi, Wang Youxin

2019, 40(2):  276-281.  doi:10.3969/j.issn.1006-7795.2019.02.022

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Objective This study aims to investigate the prevalence of suboptimal health status and its related risk factors in new undergraduates. It will provide evidence for early prevention of suboptimal health status. Methods A cross-sectional study was designed to investigate new undergraduates at Weifang university in 2017. Suboptimal Health Status Questionnaire (SHSQ-25) was used to assess the suboptimal health status of students. And all students received physical examination and blood biochemical examination. Results Totally 5 219 of 6 276 participants completed the questionnaires (87.45%). The prevalence of suboptimal health status was 8.50% in all students, 6.50% in men and 9.80% in women, respectively. Logistic regression showed that gender, exercise, eating breakfast, eating salted food, eating fried foods and sleeping quality were major influencing factors of suboptimal health status. Conclusion The prevalence of suboptimal health status in Weifang university was higher. Female, lack of exercise, less breakfast, frequent eating of pickled food, frequent eating of fried food and poor sleep quality are the influencing factors of suboptimal health status of college freshmen.

To establish abdominal wall weakness model in denervated rabbits

Yang Shuo, Cao Jinxin, Wang Minggang, Lu Jing, Chen Jie, Shen Yingmo, Zou Zhenyu, Liu Yuchen

2019, 40(2):  282-285.  doi:10.3969/j.issn.1006-7795.2019.02.023

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Objective To establish abdominal wall weakness model by motor nerve denervation in experimental rabbits. Methods Twenty experimental rabbits was randomly divided into 4 groups, one or multiple adjoining abdominal wall motor nerves were destroyed by cut off or excision of 1cm on ventral wall of rabbits:one nerve was cut off in group 1, three adjacent nerves were cut off in group 2, one nerve was excised 1cm in group 3, three adjacent abdominal wall nerves were excised 1cm in group 4. The length of bilateral abdominal wall was recorded and compared to evaluate the abdominal wall weakness and to verify the establishment of animal model. Results All experimental rabbits survived after operation, one operation wound was inflamed and recovered after 2 weeks, the length of bilatera abdominal wall had no significant change in group 1, group 2 and group 3, however, the length of abdominal wall after operation was markedly increased compared with the other side in group 4, the abdominal wall weakness appeared and the animal model was established successfully in group 4. Conclusion We successfully established the animal model of abdominal wall weakness in experimental rabbits by excising multiple adjoining abdominal wallnerves. This method is simple and feasible. It is a reliable animal model for the study of abdominal wall weakness.

Application of autoregressive integrated moving average model on monitoring influenza-like illness(ILI) case in hospital

Li Guiqin, Huang Liyong, Qin Fengzhi

2019, 40(2):  286-291.  doi:10.3969/j.issn.1006-7795.2019.02.024

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Objective To analyze the influenza-like illness (ILI) cases data reported via autoregressive integrated moving average model (ARIMA) model, and to explore the effect of the model on ILI case warning, to carry out the influenza prevention work comprehensively, and to deal with the epidemic situation efficiently.Methods The ARIMA model was established via the ILI cases data reported by Chaoyang Hospital, Capital Medical University from January 12, 2014 to October 14, 2017, and data from October 15 to December 24, 2017 was selected as the test set to evaluate predictive effect. Results The coefficient of determination (R²) of the ARIMA (2,0,0) model is 0.87. The predicted values accord well with actual values.Conclusion The model analysis of ARIMA (2,0,0) shows that it has a good early warning effect in influenza surveillance in Chaoyang Hospital, Capital Medical University. This model can provide reference for other medical institutions in monitoring ILI cases.

Study on traditional Chinese medicine treatment based on syndrome differentiation and prescription regularity of concussion practiced by Prof. Fan Yongping

Li Kangning, Fan Yongping, Wang Lei

2019, 40(2):  292-299.  doi:10.3969/j.issn.1006-7795.2019.02.025

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Objective To explore the clinical characteristics of concussion patients, traditional Chinese medicine (TCM) syndrome type distribution and prescription regularity. Methods The clinical characteristics and syndrome types of 54 patients with concussion were analyzed with the method of case analysis, and the prescription regularity of TCM for concussion were analyzed based on association rules and complex system entropy clustering algorithm. Results The ratio of male to female in 54 concussion patients was 1.5:1. The average age of the enrolled patients was (39.69±16.34) years old. Young crowd aged between 20-50 years old were in majority. Traffic accident injury was the first cause of mild traumatic brain injury. The main clinical symptoms were dizziness, headache, insomnia, forgetfulness, irritability, depression, poor appetite, nausea, and so on. Phlegm stasis obstruction was the core pathogenesis of concussion. The main TCM syndrome type included qi deficiency and phlegm stasis, heart-kidney imbalance and stasis, liver depression and phlegm stasis, and kidney deficiency and phlegm stasis. Qi deficiency and phlegm stasis was the most important syndrome type. Semen Persicae, Acorus Tatarinowii, Poria Cocos, Ligusticum Chuanxiong, Rhizoma Pinelliae(processed with alum) were the basic herbs for concussion treatment. Conclusion TCM syndrome differentiation was the key point to the concussion treatment. Phlegm stasis obstruction was the core pathogenesis of concussion. Qi deficiency and phlegm stasis was the most important syndrome type. The basic prescription was Semen Persicae, Acorus Tatarinowii, Poria Cocos, Ligusticum Chuanxiong, Rhizoma Pinelliae(processed with alum). The patients with different syndrome type were treated with the corresponding addition and subtraction of Chinese herbs in the formula according to the syndromes differentiation.

Clinical efficacy of decitabine sequential or simultaneous with “3+7” regimen for acute myeloid leukemia

Peng Xiaojuan, Zhang Ruijuan, Wang Meifang, Zhang Yaofang, Tan Yanhong, Yang Linhua

2019, 40(2):  300-304.  doi:10.3969/j.issn.1006-7795.2019.02.026

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Objective To compare the clinical efficacy and safety among decitabine combined standard induction regimen at different time points in treatment of acute myeloid leukemia(AML).Methods The clinical data of 45 AML patients enrolled from June 2012 to March 2018 were collected.The differences of clinical outcome and adverse events among the patients treated with decitabine sequentially or simultaneously with "3+7" regimen were analyzed. Results For 45 patients,the overall response rate (ORR)[complete response (CR)+CR with incomplete hematological recovery (CRi)+partial response (PR)] was 75.56%,CR/CRi was up to 55.6%(25/45);The ORR in the sequential and simultaneous group was 75.9% and 75.0%, respectively.However,there is no significant difference was observed among the two groups (P=0.842).Median followed-up time was 8 months,the overall survival(OS)of sequential group and simultaneous group was (7.25±4.61) months vs (8.48±5.42) months(P=0.447).Adverse events in the two groups were similar, mainly including cytopenia and infection,which were more common in the early stages of treatment. After the first induction treatment, the 3/4 grade myelosuppression time in the sequential group was longer than that in the simultaneous treatment group, mainly manifested as agranulocytosis, which was 15.0 d vs 8.5 d (P<0.001),and the incidence of perianal, gastrointestinal and skin and soft tissue infection was increased.The median number of suspended red blood cells and platelet infused was higher in the sequential group(P>0.05).Conclusion Whenever in sequential group or simultaneous group,the induction efficacy of decitabine combined with chemotherapy were similar,but the simultaneous group had better safety, the duration of myelosuppression was shorter, mainly manifested as shortening of the granulosis period, less blood cell infusion, and a lower incidence of infection.

Value of enhanced MRI for the evaluation of intratesticular tumors and pseudotumors

Li Min, Jiang Tao

2019, 40(2):  305-311.  doi:10.3969/j.issn.1006-7795.2019.02.027

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Objective To assess the role of 3T enhancement magnetic resonance imaging (MRI) in the prospective characterization of intratesticular tumors and pseudotumors. Methods A perspective study of 67 patients suspected of testicular disease was conducted. All patients were performed by using the 3.0T magnet.There were 65 consecutive patients with histologically confirmed, 2 patients with clinical follow-up. Via the histological characterization of testicular neoplasm as the reference standard, the diagnostic performance of MRI was compared. MR imaging allows localization of the mass (intratesticular or extratesticular)and further characterization of the lesion (solid or cystic, fat, hemorrhage, fibrosis). Gadolinium-enhanced MR imaging characteristics of tumors and pseudotumors(heterogeneous enhancement, fibrovascular septa enhancement, or no enhancement).Tumor local staging classification(invade into the tunica albuginea, epididymis, spermatic cord or scrotal wall). Results Totally 64 patients with 67 testicular lesions, 35(52.2%)of which were malignant and 32(47.8%)benign. The coherence between MRI and histologic diagnosis in differentiating benign from malignant intratesticular lesions was very well, Kappa=0.88,P value<0.001. The pathological diagnosis as the gold standard, the sensitivity of MRI in differentiating benign from malignant intratesticular lesions was 94.3%,specificity 93.8%, positive predictive value 94.3%, misdiagnosis rate 6.2%,rate of missed diagnosis 5.7%, MRI findings led to a correct histological diagnosis in 60(89.6%) of 67cases. Conclusion MRI is highly accurate in histologic characterization of testicular tumors and pseudotumors.