Journal of Capital Medical University

Dengue virus and dengue fever

Cui Xiaoyun, Wu Yanhua, An Jing

2014, 35(6): 683-688. doi:10.3969/j.issn.1006-7795.2014.06.001

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Dengue fever(DF) is the most widespread mosquito-borne diseases worldwide, caused by Dengue virus(DV). There are nearly half of the world's populations at the risk of infection in tropical and subtropical countries. DF is divided into Dengue and severe Dengue, which include Dengue hemorrhagic fever(DHF) and Dengue shock syndrome(DSS). With an estimated 500 000 cases of life-threatening disease in the form of severe Dengue every year, most of them are children. Notably, there is the most serious DF outbreak in southern China at 2014. This review will summarize several aspects of Dengue virus and Dengue fever to provide the information to the colleagues.

Sublobectomy for early stage non-small cell lung cancer

Chen Donghong, Zhi Xiuyi

2014, 35(6): 689-693. doi:10.3969/j.issn.1006-7795.2014.06.002

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Sublobectomy, include anatomic segmentectomy and non-anatomic wedge resection is an alternative to lobectomy for the surgical treatment of selected patients with early stage non-small cell lung cancer. Sublobar resection can reserve more pulmonary function in these patients. Emerging evidence suggests that sublobar resection may offer survival outcomes approaching that of lobectomy for lung cancer patients whose disease meets the following criteria: stage IA disease with no regional lymph node matastasis; tumor comes up to 2 cm in diameter; located in the periphery of lung; and predominantly ground-glass appearance on CT imaging. Compared with wedge resection, segmentectomy generated the best results. Nevertheless, the evidence is currently still limited, and the above criteria are met only in a minority of patients. Large randomized trials are underway to define the clinical role of sublobar resections, and results are eagerly anticipated. Until that time, lobectomy should still be regarded as the standard therapy for patients with early stage non-small cell lung cancer.

Antitumor activity of paclitaxel or/and cisplatin drug delivery system against lung cancer cells A549 in vitro

Cui Yong, Liu Mingliang, Wu Bingqun, Duan Xinchun, Gong Min

2014, 35(6): 694-697. doi:10.3969/j.issn.1006-7795.2014.06.003

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Objective To observe paclitaxel and/or cisplatin loaded microfiber by electrospinning technique, deliver this system to lung cancer cell A549 in vitro and observe the inhibition of cancer cell and to research effectiveness of controlled drugs delivered by electrospinning technique in antitumor field. Methods Lung cancer cell A549 was cultivated in vitro and incubated on 96-well plates with density of 1×10⁴ per well. The plates were incubated at 37 ℃ and saturated humidity for 24 hours. The plates were taken out and drugs were delivered at different concentrations in each group. There were controlled groups. Plates were incubated for 48 hours. Add in MTT(20 μL/well) and incubated for 4 hours. The medium containing MTT was discarded thoroughly and 150 μL DMSO was added, gently shaken to get a clear solution 10~15 min later. OD 490 was determined. Inhibition rate of drugs was calculated. Results Poly propylene carbonate loading paclitaxel and cisplatin controlled delivery system by electrospinning technique could inhibit cancer cell in vitro, stronger than naked paclitaxel and cisplatin and their single drug-loaded microfiber. Poly propylene carbonate loading paclitaxel or cisplatin has stronger inhibition to A549 lung cancer cells than naked paclitaxel or cisplatin. Blank poly propylene carbonate showed no inhibitory effect on the cancer cells. Conclusion Poly propylene carbonate loading paclitaxel and/or cisplatin by electrospinning technique could inhibit lung cancer cells in vitro significantly. Controlled drug-delivery system by electrospinning technique could implant antitumor drugs locally, reduce toxicity and side effect of chemotherapeutics and have a great application potential.

Preoperative CT-based pulmonary volumetric evaluation in patients with pectus excavatum: comparison with pulmonary function test

Yu Jie, Zhang Na, Chen Chenghao, Cao Juan, Zeng Qi

2014, 35(6): 698-701. doi:10.3969/j.issn.1006-7795.2014.06.004

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Objective To access the reliability of CT-based pulmonary volumetric evaluation by analyzing the correlation between lung volume calculated by CT-based pulmonary volumetric evaluation method and pulmonary function test(PFT) results. Methods This study enrolled 49 patients with pectus excavatum seen in Beijing Children's Hospital. All patients had CT scan and pulmonary function test data. The CT-scan data were used to calculate lung's volume by handdrawn-layers summation method. The lung volume and PFT result were compared. Results The lung volume calculated by CT-based pulmonary volumetric evaluation underestimated from the PFT result of VCmax, FVC and TLC. However, the correlation between lung's volume and PFT result of VCmax(P<0.001), FVC(P<0.001), TLC(P<0.001) was significant. The correlation between value of actuality/anticipated between lung volume and PFT results was also significant. Conclusion The lung volume calculated by CT-based pulmonary volumetric evaluation is significantly correlated with PFT results and this method is a useful way to evaluate the lung volume in clinical practice.

Correlation between cold ischemia time and bronchial anastomotic stenosis

Li Xin, Li Hui, Hu Bin, Hou Shengcai, Miao Jinbai, Fu Yili

2014, 35(6): 702-704. doi:10.3969/j.issn.1006-7795.2014.06.005

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Objective With the development of lung transplantation, the relationship between cold ischemia time and bronchial anastomotic stenosis requires further study. This study aimed to improve the prognosis of patients who received lung transplantation by studying the effect of cold ischemia time on bronchial anastomotic stenosis. Methods We respectively analyzed medical records of the patients, having received lung transplantation from 2009 to 2012, including the surgical records and the reports bronchoscope. Results The incidence of bronchial anastomotic stenosis of the group, of which cold ischemia time was less than 6 hours, was higher than the group of which cold ischemia time was longer than 6 hours. Conclusion The results suggest that control of lung cold ischemia time can effectively prevent the occurrence of bronchial anastomotic stenosis after lung transplantation and improve survival of patients.

Prognostic significance of preoperative serum carcinoembryonic antigen in non-small cell lung cancer: a Meta-analysis

Liu Shuku, Wang Xiaobin, Li Jie, Cao Xiaoqing, Han Yi, Liu Zhidong

2014, 35(6): 705-710. doi:10.3969/j.issn.1006-7795.2014.06.006

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Objective To explore the prognostic role of preoperative serum carcinoembryonic antigen(CEA) levels in non-small cell lung cancer(NSCLC) patients. Methods Electronic databases updated to June 1, 2014 were searched to find relevant studies. A meta-analysis was made with eligible studies which quantitatively evaluated the relationship between preoperative serum CEA level and survival of patients with NSCLC. Survival data were aggregated and quantitatively analyzed. Results Totally 16 studies(n=4 296 patients) were enrolled. We evaluated the correlation between preoperative serum CEA level and survival in patients with NSCLC. Combined hazard ratios suggested that preoperative serum CEA overexpression was associated with poor prognosis of overall survival(OS)(HR=2.28, 95% CI: 2.24-2.31) in patients. Meanwhile, for p-stage I NSCLC, the HR(95% CI) was 1.98(1.73-2.15). In the stratified analysis by patient source, significant risks were found among Asians and non-Asians. However, significant heterogeneity was observed among all studies. Conclusion In the present meta-analysis, preoperative serum CEA overexpression indicated a poor prognosis for patients with NSCLC.

Interleukin-8-251T/A gene polymorphism and lung cancer susceptibility: a meta-analysis

Zhen Dezhi, Wang Xiaobin, Li Yunsong, Li Jie, Han Yi, Liu Zhidong

2014, 35(6): 711-716. doi:10.3969/j.issn.1006-7795.2014.06.007

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Objective To perform a meta-analysis to evaluate the association between the interleukin-8-251T/A(rs4073) gene polymorphism and lung cancer risk. Methods A literature search at PubMed was performed to identify case-control study articles published before December 1st, 2013. The Stata11.0 software was used to calculate the pooled odds ratios(OR) with 95% confidence intervals(CI). And the analysis was stratified by Asian and non-Asian populations. Results Totally six articles met our inclusion criteria. The included studies yielded a total of 3 265 lung cancer cases and 3607 controls. For the homozygous A/A and A allele carriers(T/A + A/A), the pooled ORs for all studies combining 3 265 cases and 3 607 controls were 1.03(95% CI=0.92-1.14, P=0.235 for heterogeneity) and 1.07(95% CI=0.96-1.19, P=0.245 for heterogeneity) when compared with the homozygous wild-type genotype(T/T). When the analysis was stratified by Asian and non-Asian populations, significant risks were found among Asians for both the A allele carriers(OR=1.48, 95% CI=1.04-2.11, P=0.030 for heterogeneity) and the homozygous A/A(OR=1.35, 95% CI=1.02-1.92, P=0.090 for heterogeneity) individuals. However, no significant associations were found in non-Asian populations using any of the genetic models. Conclusion This Meta-analysis suggests that the interleukin-8-251 allele confer an increased risk for the development of lung cancer among Asians.

Inhibition of TRIM29 gene by siRNA suppresses the proliferation and migration of NCI-H520 lung cancer cells

Liu Chunxiao, LI Hui, Hou Shengcai, Hu Bin, Miao Jinbai, Zhang Wenqian

2014, 35(6): 717-720. doi:10.3969/j.issn.1006-7795.2014.06.008

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Objective To study the effects of TRIM29 gene on the proliferation and migration of human lung cancer cell line NCI-H520. Methods We transfected TRIM29 siRNA into NCI-H520 cells. Real time reverse transcriptase polymerase chain reaction and Western blotting assay were employed to determine TRIM29 messenger (m)RNA and protein expressions. MTT assay was used to investigate the proliferation of the cells, and the cell migration was evaluated using a transwell migration assay. Results The mRNA and protein levels of TRIM29 were significantly downregulated at 48 h after transfection in TRIM29 siRNA group compared with the control groups. The cell proliferation and migration were significantly reduced. Conclusion siRNA targeting TRIM29 gene can inhibit the proliferation and migration of human lung cancer cell line NCI-H520.

Research progress in the pathogenesis of Alzheimer's disease

Zhang Jingshuang, Wang Rong

2014, 35(6): 721-724. doi:10.3969/j.issn.1006-7795.2014.06.009

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Alzheimer's disease (AD), also known as senile dementia, is a degenerative disease of the nervous system. It is the most common type of dementia, and its incidence in the elderly is increasing all the time, the major clinical manifestations of AD are progressive memory loss, cognitive dysfunction, and other neuropsychiatric symptoms and behavioral disorders. The etiology of Alzheimer's disease is not yet clear, and its pathogenesis is complex, this article reviews briefly the past and recent important findings of potential relevance to the pathogenesis of Alzheimer's disease.

Expressions of p-JAK1 and BDNF in experimental autoimmune encephalomyelitis in mice induced by myelin oligodendrocyte glycoprotein at different doses

Qu Zhao, Zhang Li, Li Lin, Wang Qi, Yin Linlin

2014, 35(6): 725-729. doi:10.3969/j.issn.1006-7795.2014.06.010

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Objective To compare the expressions of phospho-Janus kinase 1(p-JAK1)and brain-derived neurotrophic factor(BDNF) of experimental autoimmune encephalomyelitis(EAE) in C57BL/6 mice models induced by myelin oligodendrocyte glycoprotein 35-55(MOG_35-55) at different dosages. Methods Thirty female SPF-grade C57BL/6 mice with 18-22 gram body weight were divided randomly into three groups: control group and EAE model groups(MOG_35-55 50 μg dosage group and MOG_35-55 200 μg dosage group). The mice of the two model groups were injected subcutaneously over flanks with the antigen containing 50 μg, 200 μg MOG_35-55/mouse and complete Freund's andjuvant(CFA) in the same volume, respectively. The mice of the control group were injected in the same way phosphate buffered saline(PBS) without containing MOG_35-55; 500 ng pertussis toxin(PTX) in 0.2 mL phosphate buffer solution(PBS) was given by intraperitoneal injection to the mice of the two model groups at 0 and 48 h post-immunization. The mice in control group were injected with PBS in the same way. The disease incidence, death rate, body weight and neurological score of the mice were observed. Meanwhile, the expression of JAK1 and p-JAK1 in cortex were examined by western blotting and brain-derived neurotrophic factor(BDNF) evaluated by immunohistochemical staining. Results The C57BL/6 mouse model of EAE was successfully induced by two different dosages of MOG_35-55. The expression of p-JAK1 in cortex were increased while BDNF decreased. However, the influence of MOG_35-55 200 μg dosage group on loss of weight, neurological score and the expression of p-JAK1 seemed to be more significant than MOG_35-55 50 μg dosage group. Conclusion The mouse model of immune-induced EAE was successfully established with MOG_35-55 200 μg and this EAE mouse model is stable and can be used in the drug research of multiple sclerosis(MS).

Effects of caloric restriction on learning and memory function and insulin signaling related protein in C57 mice

Zhao Zhiwei, Ma Lina, Wang Rong, Zhang Xu, Wang Yulan, Wu Yanchuan, Chen Juan, Zhang Jingyan

2014, 35(6): 730-734. doi:10.3969/j.issn.1006-7795.2014.06.011

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Objective To investigate the effect of caloric restrictionon(CR) on learning and memory function and insulin signaling related protein in C57 mice. Methods Thirty male C57 mice were randomly divided into 3 groups: normal control group, high-energy group and low energy group, each group consisted of 10 mice, learning and memory function were measured by Morris test at the sixth month. Five mouse of each group were selected to measurethe insulin signaling pathway related protein by immunochemistry. Results The escape latency of low-energy group was shorter than normal control group, which indicates the learning and memory ability of low-energy C57 mice improved. The expression of Silent mating type information regulation 2 homolog- 1(SIRT1), insulin-like growth factor 1(IGF-1), insulin receptor(IR), insulin receptor substrate-1(IRS-1), phosphatidylinositol 3-kinase(PI3K), protein kinase B(Akt/PKB), cAMP-response element binding protein(p-CREB)in brain decreased significantly as compared with normal control group(P<0.01). Conclusion Caloric restriction can improve the learning and memory ability in C57 mice, probably through downregulate the expression of insulin signaling pathway related protein in brain.

Effects of tetrahydroxystilbene glucoside on behavior and dopaminergic neurons in 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine-induced model mice

Zhang Ruyi, Zhang Li, Wu Yanchuan, Li Lin

2014, 35(6): 735-740. doi:10.3969/j.issn.1006-7795.2014.06.012

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Objective To observe effects of tetrahydroxystilbene glucoside(TSG) on behavior and content of dopamine and its metabolites in striatum of 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine(MPTP) model mice. Methods Mice were randomly divided into normal control group, model group, and TSG low dose(60 mg/kg) and high dose(120 mg/kg) groups. The behavior changes of mice were observed by pole test, Rotarod test and spontaneous movement test. The tyrosine hydroxylase(TH) positive cells were detected by immunohistochemical method. The content of dopamine(DA) and its metabolic products in striatum were determined by HPLC-ECD. Results MPTP model mice showed behavior deficit. The counts of TH positive neurons in substantia nigra and the content of dopamine and its metabolites in striatum in model mice decreased significantly compared with control group. TSG ameliorated the behavior deficits, increased the number of TH positive neurons in the substantia nigra, and elevated the content of dopamine in striatum compared with model mice. Conclusion TSG protected dopaminergic neurons against MPTP-induced damage, and may become a candidate drug for prevention and treatment of Parkinson's disease.

Changes in cognition, emotion and microglia condition caused by chronic cerebral ischemia in rats

Wang Liye, Lin Can, Zhao Haiping, Zhang Chencheng, Yan Feng, Chen Zhigang, Wang Rongliang, Wang Ningqun, Luo Yumin

2014, 35(6): 741-747. doi:10.3969/j.issn.1006-7795.2014.06.013

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Objective To observe the cognition, emotion, tissue condition and microglia condition after chronic cerebral ischemia caused by bilateral common carotid artery(BCCA) occlusion on 42nd and 56th day, and provide the basis to understand forming time of chronic cerebral ischemic model. Methods Sprague-Dawley(SD) rats were divided into 4 groups by sortition randomization method: 42 d-Sham, 42 d-Model, 56 d-Sham and 56 d-Model. Morris water maze, object location test and open field test were measured to evaluate cognition and emotion. To observe tissue condition of hippocampus, we chose H&E staining. The microglia activity was measured by Iba1 immunofluorescent staining(IF). Results The result of morris water maze showed that compared with the Sham group, the cognition of Model group was decreased, the decrease was more significant in 56 d-Model. Model group showed that the escape latency was longer, and the frequency of platform crossing was reduced. The result of object location test also showed the cognition of Model group was decreased. The discrimination index and explore time of new object was reduced. But there were no significant difference between them. The open field test showed vertical number and crossing number of Model group increased clearly, indicating that Model group had anxiety behavior. The H&E staining result showed that there were no changes in hippocampus tissue condition in the four groups. The IF staining showed that there were significant differences between the Model group and Sham group in Iba1 protein expression. Number of microglia in Model group was higher, colour was deepening, the cell was bigger, and the protuberances were increased and shortened, the protuberances like amoebocyte. Conclusion The cognition of rats was reduced at 42 days after BCCA operation, and the microglia were activated. It indicated that we can choose 42 day as forming time of chronic cerebral ischemia model. According to the cognition of 56 d-Model was worse than 42 d-Model, we suspect that the change is more significant with time. But we should notice that there were no changes in tissue condition at 42 days and 56 days described as HE staining.

Effect of local intraarterial infusion of low-dose erythropoietin in combination with tissue plasminogen activator on focal cerebral ischemia in rats

Wang Rongliang, Zhao Haiping, Wu Xiaoning, Gao Jinhuan, Ji Xunming, Luo Yumin

2014, 35(6): 748-754. doi:10.3969/j.issn.1006-7795.2014.06.014

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Objective To investigate the effect of local intraarterial infusion of low-dose erythropoietin(EPO) in combination with tissue plasminogen activator(tPA) on focal cerebral ischemic stroke. Methods Adult male Sprague-Dawley rats were subjected to middle cerebral artery occlusion for 2 or 4 h, followed by a tPA infusion over 30 min at the onset of reperfusion, with or without EPO(800 IU/kg) infusion via middle cerebral artery. A total of 50 male rats were randomly divided into 5 groups. Results Local intraarterial infusion of low-dose EPO in combination with tPA protected against transient focal cerebral ischemia in rats, manifested by a decrease in mortality, neurological deficit scores, brain edema and hemorrhage with no hematological side effects after 24 h reperfusion. The levels of p-AKT and p-ERK were both increased in response to administration of EPO, suggesting a concurrent activation of AKT and ERK pathways. Conclusion Local intraarterial infusion of low-dose EPO in combination with tPA protected against focal cerebral ischemia in rats with no significant side effects, involving activation of AKT and ERK pathways.

ACE2/ Ang-(1-7) improves glucose metabolism in HepG2 cells

Cao Xi, Yang Fangyuan, Shi Tingting, Xie Rongrong, Xin Zhong, Yang Jinkui

2014, 35(6): 755-759. doi:10.3969/j.issn.1006-7795.2014.06.015

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Objective Renin angiotensin system(RAS) is involved in glucose metabolism. This study evaluated the effect of the new pathway of RAS, ACE2/Ang-(1-7), on glucose metabolism in hepatic cells. Methods ① HepG2 cells were treated with Ang-(1-7) or A779, or transfected with ACE2 over expression plasmid DNA; ② The expression of PEPCK, G6Pase, Glut2, IRS-2, AMPKα2, SOCS-3, p22 and p67 were detected by RT-PCR; ③ ROS was measured by DHE; ④ Western blotting was used to study p22 and p67. Results ① In ACE2-overexpressing HepG2 cells, the expression of PEPCK was decreased; ② SOCS-3 was decreased, while Glut2, IRS-2 and AMPKα2 were increased significantly; ③ Ang-(1-7) reduces ROS production; ④ Ang-(1-7) depressed the expression of p22 and p67 in HepG2 cells, and ACE2 inhibited relative protein expression levels of p47^phox and p22^phox. Conclusion ACE2/Ang-(1-7) could be involved in improving glucose metabolism via anti-oxidative effects.

Effects of the gastrointestinal prokinetic agent cisapride on human ether-a-go-go related gene 2 channels expressed on HEK293 cells

Yuan Shasha, Huang Haixia, Yang Jinkui

2014, 35(6): 760-764. doi:10.3969/j.issn.1006-7795.2014.06.016

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Objective To study the effects of the gastrointestinal prokinetic agent cisapride on human ether-a-go-go related gene 2(hERG2) channels expressed on HEK293 cells. Methods hERG2 cDNA(Gene Bank accession number AF311913) was transfected transiently into the HEK293 cells using Lipofectamine 2000. HERG2 current was recorded by a whole-cell patch clamp technique before and after disposal of cisapride(0.001, 0.01, 0.1, 1.0 and 10.0 μmol/L) at the time of 2, 4, 6 and 8 min. Results The amplitude of steady-state current and peak-tail current was reduced at different cisapride concentrations. The blockage of hERG2 by cisapride is strengthened with the increasing of drug concentration at the depolarization voltage of +20 mV. The effect of cisapride is also strengthened with time, and the hERG2 current vanished at the time of 8 min with 1 μmol/L cisapride. Conclusion Cisapride blocked the hERG2 channel expressed on the HEK293 cells, including the step current and tail current. The blocking of hERG2 by cisapride is concentration dependent and time dependent simultaneously until the tail current was reduced to steady state value or disappeared completely.

Analysis of mutations in patients with Kallmann syndrome

Cao Xi, Xie Rongrong, Xin Zhong, Yang Jinkui

2014, 35(6): 765-770. doi:10.3969/j.issn.1006-7795.2014.06.017

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Objective To analyze the mutation of Kallmann syndrome 1(KAL1), prokineticin(PROK2) and prokineticin receptor 2(PROKR2) in a family, which has a 13-year-old son with Kallmann syndrome(KS). Methods ① Hormone levels were measured by immunoradiometric assay, abdomen structures were assessed by abdominal ultrasound scan, brain magnetic resonance imaging(MRI) was used to visualize the olfactory bulbs, sulci, and inner ears; ② The exon sequences of the three genes(KAL1, PROK2 and PROKR2) were screened for a mutation by direct sequencing; ③ SIFT, Polyphen and MutationTaster were used as a complementary approach to assess the significance of polymorphisms and missense mutations. Results ① The patient had sexual infantilism, and the MRI of the head revealed the dysplasia of olfactory bulbs and olfactory tract; the parents of this patient had normal pubertal development, serum gonadotropin and estradiol concentrations, and their MRI results showed normal olfactory bulbs; ② In KAL1 gene, two novel mutations(I565T and S570T) and five previously described polymorphisms(V534I, V560F, G567S, K666M and R668H) were detected in this patient. In PROKR2 gene, a homozygous Y113H was presented in this patient, and this mutation was also detected in the heterozygous state in his parents; ③ I565T, Y113H (PROKR2) and S570T may be the harmful variation according to our stringent analysis. Conclusion To date, these two novel mutations have not yet been reported in the Chinese population, and this is the first case of KS patient who exhibited two-point mutation in KAL1 and was also carried a missense mutation in PROKR2, thus indicating a possible digenic inheritance of the disease in this individual.

Dietary evaluation of hypercholesterolemia in the community

Hao Jiayin, Yu Huanling, Xiao Rong

2014, 35(6): 771-778. doi:10.3969/j.issn.1006-7795.2014.06.018

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Objective To evaluate the overall diet quality of hypercholesterolemia in the community using Chinese diet balance index(DBI-07), providing the scientific basis for community intervention. Methods Dietary information of hypercholesterolemias coming from three communities in Beijing city was obtained using Semi-Quantitative Food Frequency Questionnaire. The DBI-07 score and total energy intake, were calculated and food structure quality was evaluated according to DBI-07. Results The total dietary energy intake and protein intake ranged from(1 973.86±602.93)kcal/d to (64.40±23.99)g/d, respectively, which were lower than the RNI, while the edible oil and edible salt intake ranged from (35.57±13.90)g/d to (8.82±2.93)g/d, respectively and were much higher than the RNI. Conclusion Dietary structure of community hypercholesterolemia has an unbalanced dietary model, which is not very good for blood cholesterol management.

Comparsion of structure of protein N-glycosylation profiles between age groups

Liu Xiangtong, Wang Youxin, Yang Xinghua, Dong Jing, Wang Wei, Guo Xiuhua

2014, 35(6): 779-784. doi:10.3969/j.issn.1006-7795.2014.06.019

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Objective To explore difference of plasma N-glycosylation structure between age groups. Methods Based on cluster sampling, 212 residents of a certain community were selected and the serum N-glycans profiles were obtained by high performance liquid chromatography(HPLC).Two independent samples t test or nonparametric tests were used to compare the N-glycosylation structure between age groups divided by 35. Results The average age of the 212 subjects was(37.78±17.89) years, and the sex ratio was 0.88. More digalactosylated glycans, monosialylated glycans, core-fucosylated glycans and biantennary glycans were detected in 35 years or more, whereas more biantennary non-galactosylated glycans, disialylated biantennary glycans, trisialylated glycans and triantennary glycans were seen in younger than 35 years(P≤0.03). Conclusion The N-glycome profile in serum is age dependent. This should be taken into consideration in the development of serum glycome markers.

An efficient method of molecular cloning only by polymerase chain reaction

Xue Fenqin, Xu Qing, Wei Hua, Li Hua, Xue Bing

2014, 35(6): 785-789. doi:10.3969/j.issn.1006-7795.2014.06.020

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Objective Due to the high cost in both time and money in gene cloning, a simple molecular cloning method based on polymerase chain reaction(PCR) is presented. Methods The vector and insert are amplified by PCR separately. After DpnI digestion of the mixture of the amplified vector and insert to eliminate the DNA templates used in PCR reactions, the mixture is directly transformed into competent E. coli DH5 cells to obtain the desired clones. Results Here we report a highly simplified,reliable and efficient PCR-based cloning technique to subclone total α-synuclein gene(cDNA) from vector pET into vector pGEX-4T-1, and place total nAChRβ2 gene from vector pCDNA3.1 into vector pGEMHE. Conclusion This technique has many advantages over other cloning methods. First, we can insert any interested DNA sequences into a vector anywhere by primer designation, and it does not need to consider the restriction of multiple cloning site. Second, there is no need for any specialized enzyme digestion, gel purification of PCR product and linearized vector and enzyme ligation.

Analysis on clinicopathologic characteristics of lymphoid and hemopoietic tissue proliferation disease in orbital and ocular adnexal

He Xiaojin, Xing Li

2014, 35(6): 790-797. doi:10.3969/j.issn.1006-7795.2014.06.021

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Objective To study the clinicopathologic characteristics of lymphoid and hemopoietic tissue proliferation disease in orbital and ocular adnexal. Methods Clinical manifestations and the characteristics of clinicopathology were analyzed retrospectively in two hundreds and ten patients suffering from lymphoid and hemopoietic tissue proliferation in orbital and ocular adnexal. Results The most common histologic type in orbital and ocular adnexa is primary extraocular non-Hodgkin's lymphoma, secondly was Mikuliczs disease and inflammatory pseudotumor. There is a characteristic in age and gender among the various types of this group, and there is an overlap in clinicopathologic characteristics. The most common histologic subtype was extraocular marginal zone lymphoma of mucosa-associated lymphoid tissue type in extraocular non-Hodgkin's lymphoma, which accounted for 74.0%(74/100 cases)of cases. The extraocular marginal zone lymphoma of mucosa-associated lymphoid tissue type has different traits from that at other sites. Immunohistochemical staining is very important to the diagnosis and differential diagnosis for lymphoid and hemopoietic tissue proliferation disease in this site. The unique feature in diagnosis and clinicopathology is obvious to IgG4-related disease of this site. Conclusion We conclude that lymphoid and hemopoietic tissue proliferation disease in orbital and ocular adnexal is common among patients. There is a characteristic in age, gender and histologic subtype, which is helpful to understand these characteristics for pathological diagnosis.

Effect of promoter methylation on lower expressed cyclooxygenase-2 esophageal squamous cell carcinoma xenograft

Wang Qinggang, Meng Ying, Zhu Shengtao, Shi Haiyun, Zhang Shutian

2014, 35(6): 798-804. doi:10.3969/j.issn.1006-7795.2014.06.022

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Objective Overexpression of cyclooxygenase-2(COX-2) is known to be associated with the carcinogenesis of esophageal squamous cell carcinoma(ESCC), but in some ESCC cell lines the expression level of COX-2 may be low. In this study, methylation state of cyclooxygenase-2 gene promoter in ESCC was examined in xenografts to investigate the relationship between promoter methylation and COX-2 expression. We also studied the effect of COX-2 selective inhibitor nimesulide in conjunction with 5-aza-2 deoxycytidine on the growth of xenografts. Methods KYSE150 cell line with low COX-2 expression was used to establish ESCC xenograft in nude mice. The mice were divided into four groups, nimesulide(NIM)+5-Aza-DC(5-Aza) (group 1), NIM group(group 2), 5-Aza group(group 3), and sodium chloride control group(group 4). The growth of those xenografts in nude mice was observed. Methylation state of cyclooxygenase-2 gene promoter in xenografts was monitored using bisulfate sequencing method. RT-PCR and immunohistochemistry were employed to determine the expression level of COX-2 mRNA and protein in xenograft samples, respectively. PGE₂ concentration in xenografts was measured by ELISA. Results During the trial, mice grew well and no infection or death was observed. No difference of weight was found between groups(P>0.05). Xenograft volumes were different between groups with that of the sodium chloride group smaller than other groups, and the difference was significant(P<0.01). It was showed that methylation degree of COX-2 gene promoter was higher in NIM group and sodium choloride group than that in NIM combining 5-Aza group and 5-Aza group. For 10 CpG sites, methylation degrees were 30% and 58.3% in group 3 and group 4, respectively. The expressions of COX-2 mRNA and protein were coincidently low in group 1 and high in group 3. PGE₂ values were 0.37, 0.91 and 1.22 in group 1, 2 and 3 compared with group 4, respectively. Conclusion Promoter methylation of COX-2 gene is one of the important mechanisms to regulate COX-2 expression in low COX-2 expression ESCC xenografts. Combination of selective COX-2 inhibitor and 5-Aza has synergistic effect on ESCC xenografts.

Relationship between plasma sRAGE and heart function in patients with heart failure

Guo Xinying, Guo Caixia, Du Fenghe, Chen Buxing

2014, 35(6): 805-808. doi:10.3969/j.issn.1006-7795.2014.06.023

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Objective To investigate the level of soluble receptor for advanced glycation endproducts(sRAGE) in patients with heart failure, to analyze the relationship between sRAGE and heart function. Methods Select 100 patients with heart failure treated in our hospital(NYHA Ⅱ-Ⅳ), at the same time collect 50 cases hospitalized without heart failure, record general condition, and use the ELISA method to detect the levels of plasma sRAGE, applied SPSS 16.0 software to carry on statistical analysis. Results The plasma sRAGE level was increased in patients with heart failure than the control group, showing a significant difference(P<0.01). Plasma sRAGE level in the control group was(0.562±0.473)ng/mL, in the heart failure group was(0.844±0.705)ng/mL. With the cardiac function classification increased, plasma sRAGE level was elevated also, the control group was(0.562±0.473)ng/mL, NYHAⅡ level group was(0.680±0.506)ng/mL, NYHAⅢ level group was(0.844±0.646)ng/mL, NYHAⅣ level was(1.077±0.910)ng/mL. there is a significant difference(P=0.004). The plasma level of sRAGE had negative correlation with LVEF in patients with heart failure. Conclusion The plasma level of sRAGE was elevated in patients with heart failure, the level of plasma sRAGE and severity degree of heart failure present a associated relationship, with NYHA heart function classification increased, the plasma sRAGE level was elevated too.

Detection and significance of anti-C1q antibody in the diagnosis of children with systematic lupus erythematosus

Yin Fangrui, Pang Chunyan, Bai Li, Wang Xin, Wang Yongfu

2014, 35(6): 809-812. doi:10.3969/j.issn.1006-7795.2014.06.024

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Objective The aim of this study was to determine the associations of anti-C1q antibody with patients of children with systemic lupas erythematosus(SLE) and its potential clinica1 significance,and to explore its early clinical diagnostic significance for children with systemic lupas erythematosus. Methods There are 40 cases of SLE and 20 healthy volunteers in children were derived from the First Affiliated Hospital of Baotou Medical College from Jun.2009 to Dec.2013, the laboratory data of 40 cases of SLE were retrospectively analyzed,such as anti-SSA, anti-SSB, anti-RNP, anti-dsDNA, anti-SM, C3, C4. A method of enzyme-linked immunosorbent assay was used to detect the serum anti-C1q antibodies in 40 cases of SLE and 20 healthy volunteers, and the correlation between the anti-C1q antibodies and the laboratory data was analyzed. Results 1) The mean age was similar in children with systematic lupus erythematosus and controls(P>0.05). 2) Among 40 SLE children, the anti-C1q antibodies of 15 SLE children were positive, while the anti-C1q antibodies of 20 healthy controls were negative. Significant difference between the two groups(χ²=8.1, P<0.01).The sensitivity of anti-C1q antibodies in SLE children was 37.5% and the specificity was 100%. The sensitivity for anti-dsDNA antibodies was 62.5% and the specificity was 95%. The sensitivity for anti-SM antibodies was 22.5% and the specificity was 90%. 3) Comparing anti-C1q-positive and -negative patients, there was no difference in concentrations of anti-SSA, anti-SSB, anti-RNP, anti-Sm, C3, C4. However, there are significant differences between the two groups of anti-dsDNA antibodies(χ²=5.98, P=0.02). 4) A positive correlation was found between anti-dsDNA levels and both anti-C1q(r=0.51, P<0.01). Conclusion Our study demonstrated ClqAb has a higher sensitivity in diagnosis of SLE in children, which is helpful to the diagnosis of SLE in children with negative anti-dsDNA antibodies.

Effect of Dangguishaoyaosan on the blood lipids and the expression of inflammatory factors, such as IL-6, MCP-1, NF-κB and PPAR-γmRNA in the metaflammatory mice

Jia Lichao, Zhou Mingxue, Zhang Lei, Liu Weihong

2014, 35(6): 813-817. doi:10.3969/j.issn.1006-7795.2014.06.025

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Objective To study the effect of Dangguishaoyaosan on the blood lipids and the expression of inflammatory factors, such as IL-6, MCP-1, NF-κB and PPAR-γ mRNA in the metaflammatory mice. Methods Sixty male C57 mice were randomly divided into normal group, model group, the lipitor group and Dangguishaoyaosan group(n=15). High-fat diets joining lipopolysaccharide injection were used to build the metabolic inflammatory model in mice. Five weeks later, all groups in addition to the normal were given a gavage twice a day for five weeks,according to the dose conversed from the clinical equivalent dose. When the test was finished, we collect the blood and liver, test the serum cholesterol(TC), triglyceride(TG) and low density lipoprotein cholesterol(LDL-C) level, using flow cytometry to detect the serum level of IL-6 and MCP-1. Reverse transcription polymerase chain reaction(RT-PCR) was used to detect the expression of NF-κB and PPAR-γ mRNA in liver. Results Compared with the model group,the serum TC and LDL-C level of Dangguishaoyaosan group significantly decreased(P<0.01), and the expression of NF-κB mRNA in liver tissue decreased significantly(P<0.01), while the expression of PPAR-γmRNA significantly increased(P<0.01). Conclusion Dangguishaoyaosan can reduce the blood lipid, IL-6 and MCP-1 level of metabolic inflammation mice, and can influence metabolic inflammation in mice, which might have an intervention effect on the early atherosclerosis through regulating nuclear transcription factor NF-κB and PPAR-γ receptors.

Clinical analysis of 45 patients with geratic pancytopenia(PCP)

Zhao Jing

2014, 35(6): 818-820. doi:10.3969/j.issn.1006-7795.2014.06.026

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Objective To investigate the etiology of the geratic pancytopenia(PCP). Methods Forty-five patients with geratic Pancytopenia collected from January 2009 to January 2013 in our hospital were selected and their clinical data were retrospectively analyzed. Results Among the 45 cases of geratic Pancytopenia, 31cases were non-hematologic systemic diseases, 12 cases were hematologic systemic diseases, and the other 2 cases were of other reasons. The clinical symptoms of the non-hematologic systemic diseases were mild or moderate leucopenia and anaemia, while moderate or severe decreases of three-line blood cells happened in the hematologic systemic diseases. There was a significant difference between the decrease degree of the three-line blood cells in two kinds of diseases(P<0.05). Conclusion The geratic Pancytopenia should be paid attention to. Both hematologic systemic diseases and non-hematologic systemic diseases might be its causes. The pathogenesis should be analyzed so as to reduce the rate of misdiagnosis and missed diagnosis.

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