Table of Content

21 February 2020, Volume 41 Issue 1

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Plasma metabolomic profiling of proliferative diabetic retinopathy

Zhu Xiaorong, Yang Fangyuan, Lu Jing, Cao Xi, Yang Guangran, Xie Rongrong, Feng Jianping, Yang Jinkui

2020, 41(1):  1-7.  doi:10.3969/j.issn.1006-7795.2020.01.001

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Objective To investigate the plasma "metabolic fingerprints" of proliferative diabetic retinopathy (PDR) and to explore associated pathogenesis. Methods A total of 1 024 patients with type 2 diabetes mellitus were screened. To match clinical parameters between the case and control subjects, patients with PDR (n=21) or those with a duration of diabetes of ≥ 10 years but non-diabetic retinopathy (NDR, n=21) were assigned to the present case-control study. Distinct metabolite profiles of serum were examined using liquid chromatography-mass spectrometry (LC-MS). Results A total of 136 distinct metabolites between PDR and NDR groups were identified. These metabolites mainly included organic compounds (78%), organoheterocyclic compounds (4%), lipids and lipid-like molecules (3%) and others. Altered metabolites were enriched in 30 KEGG pathways. Three of them were significantly enriched (P<0.05), namely, sulfur metabolism, sphingolipid metabolism, cysteine and methionine metabolism. Conclusion We generated a metabolomic profile for extreme eye phenotype between PDR and NDR groups. The impairment in the metabolism of sulfur, sphingolipid, cysteine and methionine were identified as metabolic dysregulation associated with PDR, which might provide insights into potential new pathogenic pathways for diabetic retinopathy.

Construction and preliminary application of conditional KCNH6-knockout mice

Lu Jing, Li Qi, Zhu Xiaorong, Xie Rongrong, Yang Jinkui

2020, 41(1):  8-13.  doi:10.3969/j.issn.1006-7795.2020.01.002

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Objective To construct KCNH6 gene specific liver knockout mice and observe the changes of liver lipid metabolism. Methods KCNH6^flox/flox transgenic mice were constructed by Cas9 technique and hybridized with Alb-cre mouse which was specifically expressed Cre in liver cells. The genotype was identified by polymerase chain reaction (PCR). Weight and food intake in knockout mice and wildtype mice were detected. Triglycerides (TG), total cholesterol (TC), high density lipoprotein-cholesterol (HDL-C) and low density lipoprotein-cholesterol (LDL-C) were assessed at 8 and 20 weeks, respectively. Results KCNH6 gene liver cell specific knockout mice were constructed, with the genotype of KCNH6^flox/flox/Cre^T. There were no significant changes in body weight and food intake of the mice in the control group. There was no significant change in lipid metabolism in male and female mice at 8 weeks. Cholesterol levels increased in male mice at 20 weeks. Conclusion The mouse model of KCNH6 gene liver cell specific knockout was constructed. Abnormal lipid metabolism was observed. The establishment of animal model provides a research platform to explore the role of KCNH6 gene in liver lipid metabolism.

Generation and identification of Unc13 gene pancreatic beta cell knockout mice model

Li Qi, Lu Jing, Zhu Xiaorong, Xiong Fengran, Yang Jinkui

2020, 41(1):  14-20.  doi:10.3969/j.issn.1006-7795.2020.01.003

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Objective To construct Unc13 gene pancreatic beta cell knockout mice by the Cre recombinase system for the study of the role and mechanism of Unc13 in insulin secretion. Methods Unc13flox/flox transgenic mice were constructed by CRISPR/Cas9 technology and hybridized with pancreatic beta cell specific Cre recombinase (Ins2-cre)tool mice. The progeny genotype was identified with PCR and sequencing technology. The knock out (KO) and their wild type (WT) mice were used to measure body weight, glucose tolerance, and insulin secretion level. Results The pancreatic beta cell specific Unc13 gene conditional knockout mice (hereinafter referred to as Unc13 KO mice) were constructed. Further studies showed that Unc13 KO mice had impaired glucose tolerance and decreased first phase insulin secretion but no significant change in body weight compared with WT mice. Conclusion The Unc13 KO mice model was constructed, which provides a research platform for exploring the role of Unc13 gene in the development of diabetes.

Effects of storage condition on measurement of haptoglobin in urine

Zheng Xiaomin, Yang Fangyuan, Liu Cuiping, Yang Jinkui

2020, 41(1):  21-26.  doi:10.3969/j.issn.1006-7795.2020.01.004

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Objective To investigate the effects of different treatments of urinary specimen and storage conditions on urinary haptoglobin levels. Methods The urinary samples of 40 patients with type 2 diabetes were stored at room temperature and 4℃ for 3 hours, 6 hours and 24 hours with and without centrifugation, and then the levels of urinary haptoglobin were measured with enzyme-linked immunosorbent assay. Results For the samples without centrifugation, no change in urinary haptoglobin concentration was observed after 3 hours and 6 hours of room temperature storage(both P>0.05), but there was a significantly decrease after 24 hours(P < 0.01). There was no statistically significant difference in the levels of urinary haptoglobin after 3 hours at 4℃ (P>0.05), and there was a significantly decrease after 6 hours(P < 0.01). The centrifuged samples were stable for 3 hours at both room temperature and 4℃(both P>0.05), but there was a significantly decrease after 6 hours(both P < 0.01).Conclusion It was recommended to store urine samples at room temperature when the measurement of urinary haptoglobin could be completed within 6 hours. Cold storage and centrifugation resulted in the decrease of urinary haptoglobin level.

Relations between infected with Helicobacter pylori and diabetic nephropathy in the patients with newly-diagnosed type 2 diabetes mellitus

Li Bin, Cui Limei, Lyu Naqiang, Yan Yehua, Zheng Xiaomin, Tao Jing, Lui Cuiping

2020, 41(1):  27-30.  doi:10.3969/j.issn.1006-7795.2020.01.005

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Objective To study the relation between Helicobacter pylori(HP) infection and risk of diabetic nephropathy(DN) in the newly-diagnosed type 2 diabetes mellitus(T2DM) patients. Methods Totally 371 patients with the newly-diagnosed T2DM were enrolled between May 2014 and May 2018 at Department of Endocrinology, Beijing Chuiyangliu Hospital.According to urinary microalbumin to creatine ratio(UACR), these patients were divided into three groups:DN0 group (n=215), with UACR (male <2.5 mg/mmol and female<3.5 mg/mmol); DN1 group (n=128), with UACR (male 2.5 mg/mmol ≤ UACR < 30 mg/mmol, female 3.5 mg/mmol ≤ UACR < 30 mg/mmol); DN2 group(n=28), with UACR ≥ 30 mg/mmol.Results Single factor analysis showed that HP infection was associated with DN microalbuminuria (OR=1.77, 95% CI:1.103-2.851), but HP infection wasn't associated with DN clinical proteinuria.The Logistic regression analysis showed that the risk of DN was 2.52 fold higher in newly-diagnosed type 2 diabetic patients with positive HP than those with negative HP(OR=2.516, 95%CI:1.305-4.849).Conclusion HP infection is related to DN in newly-diagnosed T2DM patients. HP infection can increase the risk of DN in T2DM patients.

Effect of angiopoietin-like 4 in the diagnosis of nonalcoholic fatty liver

Yang Longyan, Wang Yan, Xu Yuechao, Cheng Yanan, Ma Yan, Zhao Dong

2020, 41(1):  31-34.  doi:10.3969/j.issn.1006-7795.2020.01.006

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Objective To study the relationship between angiopoietin like protein 4 (ANGPTL4) and non-alcoholic fatty liver disease (NAFLD), and to explore the value of ANGPTL4 in the diagnosis of NAFLD. Methods Totally 52 patients with NAFLD and 47 normal controls were selected from Beijing Luhe Hospital, Capital Medical University from January to December 2018. All subjects were examined by the same ultrasound doctor. The patients were diagnosed with mild and moderate to severe NAFLD. Patients with diabetes, coronary heart disease, cerebrovascular disease, chronic liver disease, taking lipid-lowering drugs or other drugs are excluded. The level of serum ANGPTL4 was measured with enzyme linked immunosorbent assay (ELISA). The difference of ANGPTL4 between the control and NAFLD groups, as well as the patients with mild, moderate and severe NAFLD was analyzed. The relationship between ANGPTL4 and NAFLD was analyzed with clinical case-control study. The possibility of ANGPTL4 as a potential serum molecular marker for NAFLD was analyzed with ultrasonic diagnosis as the gold standard. Results Compared with the control group, the level of serum ANGPTL4 in patients with NAFLD increased and tended to increase with the severity of NAFLD (F=4.553, P=0.035 4). The area under the (receiver operating characteristic curve, ROC) curve of ANGPTL4 was 0.638, and the diagnostic threshold of NAFLD was determined with the maximum Jordan index. The optimal threshold was 90.93 ng/mL. The results of this standard were compared with those of ultrasonic diagnosis. Sensitivity=100%, specificity=46.8%, kappa=0.48. Conclusion The serum ANGPTL4 level was related to NAFLD and its severity, but the misdiagnosis rate of clinical assistant diagnosis of NAFLD was high.

Clinical characteristics and risk factors of obstructive sleep apnea syndrome in patients with type 2 diabetes mellitus

Wei Jiajie, Jiao Jie, Zhou Yingsheng

2020, 41(1):  35-39.  doi:10.3969/j.issn.1006-7795.2020.01.007

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Objective To study the clinical characteristics and risk factors of obstructive sleep apnea syndrome(OSAHS) in patients with type 2 diabetes mellitus(T2DM), and analysis the correlation. Methods Patients were divided into OSAHS group(225 cases) and non-OSAHS group (76 cases) in Beijing Anzhen Hospital, Capital Medical University from 2012 to 2019.Height, body weight, body mass index (BMI), waist circumference(WC), systolic blood pressure(SBP) and diastolic blood pressure (DBP) were measured in all cases. Fasting plasma glucose(FPG), hemoglobin A1c (HbA1c), blood lipid, uric acid(UA), liver and kidney function indexes were tested. Result Compared with the non-OSAHS group, WC, BMI, UA, and HbA1c levels were significantly increased (P<0.05 or 0.01). Compared with the non-OSAHS group, the prevalence of hypertension and coronary heart disease was significantly increased (P<0.05), and high density lipoprotein cholesterol(HDL-C) was significantly decreased (P<0.01). Multivariate Logistic analysis was performed with OSAHS in T2DM patients as the dependent variable. The results showed that the history of coronary heart disease, WC, UA, HDL-C and HbA1c were the risk factors for T2DM combined with OSAHS. Conclusions T2DM patients have a history of coronary heart disease, increased waist circumference, high uric acid level, low HDL-C level and high HbA1c, which increase the risk of OSAHS.

A study of metabolic syndrome and cardiovascular disease risk status in highly active antiretroviral therapy-naive patients with human immunodeficiency virus infection

Wang Yanxue, Hu Hongying, Li Xingang, Lu Xingmeng, Gao Guiju, Liang Hongyuan, Xiao Jiang, Yang Di, Wang Fang, Han Xiaotao

2020, 41(1):  40-44.  doi:10.3969/j.issn.1006-7795.2020.01.008

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Objective To investigate the clinical characteristics as well of metabolic syndrome(MS) and risk status of cardiovascular disease in highly active antiretroviral therapy(HAART)-naive patients with human immunodeficiency virus(HIV) infection.Methods With 193 HIV infected patients as the study subjects, patients with(case group)and without(control group)MS were identified according to the MS diagnostic criteria, According to the Framingham risk score, patients with high risk of cardiovascular disease were selected as middle and high risk group, and those with low risk were selected as low risk group. Clinical characteristics were analysed by comparing the two groups. Results In our research, the prevalence of MS was 9.33%. The largest component was body mass index (BMI) ≥ 25 kg/m², followed by a decrease in high density lipoprotein cholesterol(HDL-C).Compared with the control group, patients with MS were older, and they had larger body mass index (BMI) (P < 0.05). The cardiovascular risk of MS patients was significantly higher than those without MS.Age, BMI and smoking history were associated with cardiovascular risk in newly diagnosed HIV patients. Patients at high risk of coronary heart disease had higher HIV RNA load, but the difference was not statistically significant.Conclusion Age and BMI were associated with the risk of metabolic syndrome.Cardiovascular risk of MS patients was significantly higher than those without MS.Related risk factors should be evaluated before HAART, and metabolic disorders and cardiovascular diseases should be closely monitored.

Screening of glucokinase gene in pregnancy women with abnormal glucose metabolism

Yuan Shasha, Ma Yan, Wu Nannan, Xu Yuechao, Yang Longyan, Ke Jing, Zhao Dong

2020, 41(1):  45-49.  doi:10.3969/j.issn.1006-7795.2020.01.009

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Objective To estimate the prevalence of maturity onset diabetes of the young 2 (MODY2) caused by glucokinase (GCK) gene mutation and to establish a clinical strategy for identifying GCK gene mutation in pregnancy woman. Methods A retrospective study was conducted on pregnant women undergoing pregnant examination at Luhe Hospital, Capital Medical University on January 1, 2016 and December 31, 2018.The GCK gene was sequenced in patients meeting the following criteria:fasting blood glucose(FBG) ≥ 5.5 mmol/L (at least 3 times) and triglyceride (TG) ≤ 1.43 mmol/L (at least once). The 2-hour increment of 75 g glucose tolerance test was less than 4.6 mmol/L in 24-28 weeks of gestation. Results A total of 2 454 subjects were screened of which 12 patients met the screening criteria. DNA samples of 5 patients were sequenced, with 1 patient with GCK mutation identified. Conclusion Our screening method based on FBG ≥ 5.5 mmol/L (at least 3 times), triglyceride ≤ 1.43 mmol/L(at least once) and oral glucose tolerance test(OGTT)2 h glucose increment < 4.6 mmol/L is feasible for screening GCK gene mutation in pregnant women. GCK mutation is not the main cause of abnormal gestational blood glucose in China.

Current status of research concerning animal models of cognitive impairment of schizophrenia

Liu Xinyao, Ma Chao, Jia Hongxiao

2020, 41(1):  50-54.  doi:10.3969/j.issn.1006-7795.2020.01.010

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The mechanism of schizophrenia is not yet precise. Cognitive impairment is a difficult point in the treatment of schizophrenia. Reliable and predictable animal models could improve understanding of the neurobiological basis of the disease, and also is crucial for developing new drugs that work better. This review will focus on pharmacological models, neurodevelopmental models and genetic models and provides ideas for studying and alleviating the cognitive impairment of schizophrenia.

Clinical changes of cognitive function patients with spleen deficiency

Wu Fengzhi, Ning Yanzhe, Jia Hongxiao

2020, 41(1):  55-58.  doi:10.3969/j.issn.1006-7795.2020.01.011

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Objective To study the clinical characteristics of cognitive function change in patients with spleen deficiency(PSD). Methods Twenty six subjects with spleen deficiency syndrome and 25 healthy subjects were measured by Clinical Memory Scale Test(CMS) and Continue Performance Test(CPT), which were used to assess memory and attention. Results Compared with healthy controls, patients with spleen deficiency syndrome showed significantly lowered scores in the directed memory(t=-3.60, P=0.001), associative memory(z=-3.47, P=0.001)and meaningless image recognition(t=-2.09, P=0.037), but high in the scores of 2-FT(Z=3.13, P=0.002)and 3-FT(Z=2.45, P=0.014). Conclusion Our findings suggest that patients with spleen deficiency syndrome showed impaired memory and attention, which provide new evidence to support treating cognitive disorder from spleen.

Cognitive impairment in patients with depersonalization disorder and its correlation with clinical symptoms and social function

Song Nan, Jia Hongxiao

2020, 41(1):  59-63.  doi:10.3969/j.issn.1006-7795.2020.01.012

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Objective To study the impairment of cognitive function in patients with depersonalization disorder and its correlation with clinical symptoms and social function. Methods Forty eight who meet the diagnostic criteria in the Diagnostic and Statistical Manual of Mental Disorders (Fifth Edition, DSM-V) and the Cambridge Depersonalization Scale (CDS) score of ≥ 70 patients with personality disintegration disorder and 32 healthy subjects used Trail Making Test A (TMTA), symbolic coding, Hopkins Verbal Learning Test Revised (HVLT-R), Brief Visuospatial Memory Test Revised (BVMT-R), Continuous Performance Test (CPT) and Stroop Color Test for cognitive function assessment, and collect general demographic and clinical information on relevant information perform analysis. Results There were significant differences in cognitive function of TMTA (P=0.03), HVLT-R (P=0.01), BVMT-R (P=0.01), and Stroop test (P=0.01) between the depersonalized patients and the healthy controls; There was no statistically significant difference in cognitive function between the medication group and the non-medication group in patients with disintegrated personality (P>0.05). There was a statistically significant difference in Global Assessment Function(GAF) scores on social function between the depersonalized patient group and the healthy test group (P=0.00); Stroop test and CPT test in cognitive function were negatively correlated with clinical symptoms (r=-0.455, P<0.05; r=-0.292, P<0.05), and TMAT test was negatively correlated with social function (r=-0.300, P<0.05), the Stroop test and the BVMT-R test were positively correlated with social function (r=0.352, P<0.05; r=0.344, P<0.05), but the relationship was weak. Conclusion Patients with personality disintegration have cognitive impairment and social function decline. Attention/alertness in cognitive function is negatively correlated with clinical symptom scores. There is a correlation between cognitive function and social function. Impaired cognitive function can lead to social function decline.

Analysis of effect of Shi-Zhen-An-Shen-Tang on intestinal microflora diversity of mice exposed to cuprizone based on 16S rRNA technique

Ma Chao, He Yi, Sun Zuoli, Liu Xinyao, Feng Zhengtian, Chen Pei, Ning Yanzhe, Zhu Hong, Yin Dongqing, Jia Hongxiao

2020, 41(1):  64-70.  doi:10.3969/j.issn.1006-7795.2020.01.013

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Objective To explore the effect of Shi-Zhen-An-Shen-Tang(SZAST) on intestinal microflora diversity of demyelinated model mice, and further to reveal its therapeutic characteristics and mechanisms based on the 16S rRNA technique. Methods Thirty Young adult male C57BL/6 mice (5 weeks old) were randomly divided into six groups, each group of 5 mice:normal control, model, and low dosage of SZAST, medium dosage of SZAST, high dosage of SZAST and quetiapine(QTP)-treated group. The control group were given mixed cuprizone (CPZ, a copper chelator, 0.2%, w/w) rodent chow for six successive weeks to induce demyelination. During the last two weeks, mice were given an oral gavage of saline, or SZAST of three different doses (a low dose of 5.5 g·kg^-1·d^-1, a medium dose of 11.0 g·kg^-1·d^-1, or a high dose of 16.5 g·kg^-1·d^-1), or QTP, respectively. Then, the mice were anesthetized and their cecum contents removed. Illumina MiSeq was used as the sequencing platform to analyze and evaluate the number of operational taxonomic units (OTUs), richness and diversity indexes, diversity of alpha and beta, differential phylum and genus of intestinal flora in V3-V4 zone of 16S rRNA. Results SZAST could regulate the number of OTUs, richness and diversity indexes of intestinal flora, imbalance of alpha and beta diversity of demyelinated model mice.Compared to the control group, Lachnospiraceae family were decreased in the model group (p<0.05), and SZAST could increase the diversity of Lachnospiraceae family. Compared to the control group, Beta-proteobacteria, Burkholderiales, Alcaligenaceae, and Bacteroidales were increased in the model group (p<0.05), but SZAST could decrease these intestinal flora. Conclusion SZAST had a therapeutic effect on the abnormal flora diversity of demyelinated model mice. It revealed the relevant pathological mechanism of mental schizophrenia caused by white matter damage through 16S rRNA technology.

Study on attention networks in patients with shift work sleep disorder

Liu Yulian, Jia Hongxiao, Song Nan, Yao Hao, Ning Yanzhe

2020, 41(1):  71-74.  doi:10.3969/j.issn.1006-7795.2020.01.014

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Objective To study the impairment of attention networks in patients with shift work sleep disorder (SWSD). Methods A total of 30 patients with SWSD and 26 matched healthy subjects were recruited to undergo attention network test (ANT). Results Compared with healthy subjects, the patients with SWSD showed significantly poorer efficiency on alerting attention and executive attention(P < 0.01).Furthermore, efficiency on executive attention was positively associated with the scores of Pittsburgh sleep quality index (PSQI). Conclusion Our findings suggest that patients with SWSD exhibited selective impairments of attention network of alerting and executive control.

Clinical observation of abdominal acupuncture in the treatment of obesity caused by atypical antipsychotics

Li Xue, Zhu Hong, Fang Meng, Ning Yanzhe, Zhao Shuang, Yin Dongqing

2020, 41(1):  75-79.  doi:10.3969/j.issn.1006-7795.2020.01.015

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Objective To investigate the effects of abdominal acupuncture on body weight, waist circumference, body mass index (BMI), and blood lipids in obese patients with atypical antipsychotics.Methods Sixty patients with obesity caused by atypical antipsychotics were divided into a treatment group (30 cases) and a control group (30 cases). The treatment group was treated with abdominal acupuncture, while the control group with health guidance. The changes of body weight, waist circumference, BMI and blood lipid levels of patients in the two groups before treatment were observed, on the 28th day and on the 56th day.Results Repeated multivariate analysis of variance was used to study the results. The results showed that the time effects and grouping effects on weight, waist circumference, BMI, cholesterol and triglycerides were significantly interactive (P<0.05). Further analysis indicated that the waist circumference and BMI data on the 28th day of treatment were better than the control group (P<0.05). On the 56th day, the weight, waist circumference, BMI, and triglyceride data of the treatment group were better than the control group (P<0.05), with the statistically significant difference. Conclusion Abdominal acupuncture can decrease the body weight, BMI, and lipid metabolism of obese patients with antipsychotics.

FSTL1 promotes proliferation, adhesion and tube formation of pulmonary artery endothelial cells

Sha Yuhui, Gao Yang, Liu Jie, Qi Xianmei, Han Lulu, Wang Wang

2020, 41(1):  80-86.  doi:10.3969/j.issn.1006-7795.2020.01.016

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Objective To investigate the effect of follistatin-like 1(FSTL1) on human pulmonary artery endothelial cells (HPAECs) and to replenish the role of FSTL1 in pulmonary hypertension (PH). Methods The levels of FSTL1 in HPAECs were evaluated by qRT-PCR and Western blotting. Cellular viability was determined by MTT after giving exogenous recombinant human FSTL1 or small interfering RNA (siRNA). We also observed the effect of FSTL1 on adhesion and angiogenesis in HPAECs. Results Exposure to hypoxia upregulated the expression of FSTL1 mRNA and protein in HPAECs (P<0.01). The cellular viability of HPAECs stimulated by FSTL1 250 μg/L and 500 μg/L were higher than FSTL1 0 μg/L group under normoxia (P<0.01). SiRNA-mediated knockdown of FSTL1 attenuated hypoxia-induced proliferation of HPAECs compared with the negative control group (P<0.001). The amount of adherent cells stimulated with FSTL1 250 μg/L was increased under normoxia compared with that in the control group (P<0.001). The total length and the amount of tubes formed by HPAECs stimulated with FSTL1 250 μg/L were both increased under normoxia compared with the control group (P<0.001). Conclusions FSTL1 promotes proliferation, adhesion and tube formation of HPAECs.

Sensitivity study of HT22 in ferroptosis

Yang Tianli, Yang Yongfei, Yuan Zengqiang

2020, 41(1):  87-91.  doi:10.3969/j.issn.1006-7795.2020.01.017

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Objectives To investigate whether HT22 cell line is a good cell model to study the ferroptosis and its molecular mechanisms. Methods HT22 cells were treated with Erastin or RSL3, the putative ferroptosis inducers, followed by the observation of morphology change under microscope. Moreover, cell viability was measured by cell counting kit 8 (CCK-8), and cellular lipid peroxidation was measured by MDA assay and the cellular concentration of Fe²⁺ was measured by Fe assay. In addition, the mRNA expression levels of SLC7A11 and PTGS2 were measured by quantitative real time polymerase chain reaction(qRT-PCR). Results Erastin or RSL3 treatment significantly increased lipid peroxidation in HT22, which lead to ferroptosis. However, Erastin or RSL3 did not alter the Fe²⁺ concentration in HT22 cells. In addition, we found that Erastin or RSL3 transcriptionally upregulated the expressions of SLC7A11 and PTGS2. Conclusion HT22 is a suitable cell model to study ferroptosis and the underlying mechanism of the accumulation of lipid peroxidation during ferroptosis.

Effect of Snail-mediated lung epithelial-mesenchymal transition in activation of myofibroblasts

Li Siling, Zhu Zhonghui, Li Qiuyue, Xu Chunjie, Zhao Jing, Wang Yan, Tian Lin

2020, 41(1):  92-98.  doi:10.3969/j.issn.1006-7795.2020.01.018

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Objective To explore the effect of nuclear transcription factor Snail-mediated lung epithelial-mesenchymal transition (EMT) in activation of myofibroblasts. Methods MLE-12 cells (murine epithelial cell line) were stimulated by transforming growth factor-β1 (TGF-β1) to build EMT model, the mRNA expression of epithelial marker CDH1, mesenchymal marker vimentin and nuclear transcription factor Snail were detected by RT-qPCR. MLE-12 cells were co-cultured with mouse embryo fibroblast (NIH-3T3) cells. The mRNA expression of α-smooth muscle actin (α-SMA), collagen Ⅰ α1(COL1A1), collagen Ⅲ α1(COL3A1) of NIH-3T3 were detected by RT-qPCR. MLE-12 cells were transfected with Snail-shRNA lentivirus. The expression of Snail mRNA and protein of MLE-12 cells were detected by RT-qPCR and Western blotting. The Snail-shRNA-transfected-MLE-12 cells were co-cultured with NIH-3T3 cells. The mRNA expression of α-SMA, COL1A1, COL3A1 of NIH-3T3 were detected by RT-qPCR. Multiple groups were analyzed by one-way ANOVA and LSD test to determine significant differences between groups at P<0.05. Results The results of RT-qPCR showed that after stimulated by TGF-β1 for 48 hours, compared with control, the mRNA expression of CDH1 of MLE-12 in TGF-β1 group was down-regulated while α-SMA and Snail were up-regulated (P<0.05). In the co-culture model, the results of RT-qPCR showed that compared to TGF-β1 and MLE-12 group, MLE-12 stimulated by TGF-β1 cause up-regulation of α-SMA, COL1A1, COL3A1 mRNA expression of NIH-3T3 (P<0.05). After transfected with Snail-shRNA lentivirus, mRNA and protein expression of MLE-12 were down-regulated compared with control virus group (P<0.05). In the co-culture model of lentivirus-transfected-MLE-12 and NIH-3T3, results of RT-qPCR showed that α-SMA, COL1A1, COL3A1 mRNA up-regulation of NIH-3T3 in TGF-β1+Snail-shRNA group were lower than TGF-β1+control-shRNA group (P<0.05). Conclusion Snail-mediated EMT of MLE-12 can cause activation of myofibroblasts, knockdown of Snail in MLE-12 can inhibit fibroblasts activation into myofibroblasts, suggested Snail-mediated lung epithelial-mesenchymal transition plays an important role in myofibroblasts activation.

Associations of plasma trimethylamine N-oxide and its precursors with coronary heart disease

Xiong Xiaowei, Guo Caixia, Xu Xiaowei, Mu Liying, Hu Hongyu, Chen Wei, Chen Buxing

2020, 41(1):  99-102.  doi:10.3969/j.issn.1006-7795.2020.01.019

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Objective To study the associations of plasma trimethylamine N-oxide (TMAO) and its precursors with coronary heart disease (CHD). Methods Patients with suspected CHD were enrolled in cardiology department of Beijing Tiantan Hospital, and divided into CHD and control groups according to angiography results. Plasma TMAO, choline, carnitine, betaine and butyrobetaine of all patients were measured by high-performance liquid chromatography, and the differences of plasma TMAO and its precursors between CHD and control patients were analyzed by statistical methods. Results The difference of plasma TMAO between CHD and control groups was significance[(6.20±3.57)μmol/L vs (3.90±2.26) μmol/L, P<0.05], but the differences of choline, carnitine, betaine and butyrobetaine between two groups were not significant (P>0.05). And plasma TMAO was still associated with CHD significantly in multivariate Logistic regression analysis (OR=1.35, P<0.05). Conclusion CHD was associated with plasma TMAO level significantly, but not its precursors.

Correlation between TMPRSS2-ERG fusion gene and survival rate of metastatic castration-resistant prostate cancer after chemotherapy

Xie Yingwei, Jin Shipeng, Li Shuang, Wang Yonghui, Wang Wei, Ping Hao, Liu Yuexin

2020, 41(1):  103-107.  doi:10.3969/j.issn.1006-7795.2020.01.020

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Objective To determine the correlation of TMPRSS2-ERG(T-E) fusion gene to the survival rate in metastatic castration-resistant prostate cancer after docetaxel chemotherapy. Methods We included 20 patients with benign prostatic hyperplasia and 50 patients with prostate cancer(mCRPC) chemotherapy. The expression of T-E fusion gene in peripheral blood mononuclear cells was examined by quantitative real time polymerase chain reaction(qRT-PCR). Therapeutic responses of patients with different fusion gene expression were assessed by prostate specific antigen (PSA) response rate, PSA-progression free survival(PFS), radiological progression free survival(RX-PFS), and overall survival(OS). Results The T-E fusion gene was detected in 10 (20%) of 50 prostate cancer patients. The PSA response rate in the T-E positive group was lower than that in the negative group (32.52% vs 74.35%, P=0.040). Univariate survival analysis showed that PSA-PFS, RX-PFS, and OS were lower in T-E positive patients than in negative patients, and the difference was statistically significant (P<0.01). Multivariate Cox regression analysis showed that T-E positive was an independent risk factor for PSA-PFS, RX-PFS and OS (P=0.010, P=0.010, P=0.020). Conclusion This study demonstrates that T-E fusion gene expression is an independent risk factor for PSA-PFS, RX-PFS and OS in mCRPC patients treated with docetaxel.

Risk factors for osteoporosis in patients with metabolic syndrome in old people with type 2 diabetes mellitus

Xiu Shuangling, Mu Zhijing, Sun Lina, Zhao Lei, Han Qing, Cao Xuexia

2020, 41(1):  108-112.  doi:10.3969/j.issn.1006-7795.2020.01.021

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Objective To explore the risk factors for osteoporosis in patients with metabolic syndrome in old people with type 2 diabetes mellitus. Methods A total of 301 old people with metabolic syndrome and type 2 diabetes aged more than 60 years-old were included in the study. Levels of hemoglobin A1c(HbA1c), fasting insulin (FINS), vitamin D, blood lipid, serum C-reactive protein (CRP)and interleukin-6 (IL-6) were measured. Body mass index (BMI) was calculated. Furthermore, bone mineral density (BMD), skeletal muscle mass and grip strength were assessed. The risk factors for osteoporosis were evaluated. Results Compared with people without osteoporosis, people with osteoporosis had lower levels of BMI, vitamin D, skeletal muscle mass index and grip strength (P < 0.05). In a multiple stepwise Logistic analysis, being female and lower grip strength were associated increased risk of osteoporosis (OR=2.85, P < 0.001; OR=2.34, P < 0.001, respectively), while higher BMI was associated with decreased risk of osteoporosis (OR=0.84, P < 0.001).Conclusion Being female and lower grip strength were independent risk factors for osteoporosis in patients with metabolic syndrome in old people with type 2 diabetes mellitus, while higher BMI was a protective factor.

Effects of combined sacubitril/valsartan and metoprolol succinate sustained release tablets on patients who suffer from the coronary heart disease with chronic heart failure

Xia Yong, Zhang Jianqing, Wang Shaoxia, Chen Dan, Ji Jianmin

2020, 41(1):  113-118.  doi:10.3969/j.issn.1006-7795.2020.01.022

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Objective This paper was to explored the clinical efficacy of combined sacubitril/valsartan and metoprolol succinate sustained release tablets on patients who had the coronary heart disease with chronic heart failure. Methods This paper selected patients who suffered from the coronary heart disease with chronic heart failure in The Third Affiliated Hospital of Henan University of Science and Technology and Zhengzhou Cardiovascular Hospital from September 2017 to September 2018 as the samples. Based on intergroup equilibrium, the samples were divided into treatment group (n=94) and control group (n=93) uniformly by hospitalization date, age, disease course, and gender. Metoprolol succinate sustained release tablets were applied in control group, while sacubitril/valsartan and metoprolol succinate sustained release tablets were combined in treatment group. The therapy for patients in both group lasted for 6 months. This paper compared the clinical efficacy and cardiac function index:maximum early filling speed (E), left ventricular ejection fraction (EF), maximum filling speed in late diastole (A), E/A, vascular endothelial function[plasma endothelin (ET), calcitonin gene-related peptide (CGRP), nitric oxide (NO)], between two groups before and after the therapy. Results The clinical efficacy in treatment group was 93.62% which was significantly 78.49% higher than that in control group. After the therapy, E, EF, E/A, CGRP, NO in treatment group were higher than those in control group, while A and ET were lower in treatment group (P<0.05). Conclusion The therapy of combined sacubitril/valsartan and metoprolol succinate sustained release tablets had significant efficacy on coronary heart disease and heart failure by improving the cardiac function.

Relationship between sarcopenia and bone mineral density in middle-aged and aged men with type 2 diabetes mellitus

Li Yaao, Xiu Shuangling, Wang Li

2020, 41(1):  119-124.  doi:10.3969/j.issn.1006-7795.2020.01.023

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Objective To explore the relationship between sarcopenia and bone mineral density (BMD) in middle-aged and aged men with type 2 diabetes mellitus (T2DM). Methods All 218 men with T2DM aged over 55 years were selected from the Department of Endocrinology of Xuanwu Hospital. The BMD of lumbar spine and hip, appendicular skeletal muscle mass (ASM), appendicular fat mass (AFM), appendicular skeletal muscle mass index (ASMI), body mass index(BMI) were measured. The association between sarcopenia and osteoporosis was assessed. Results The prevalence of sarcopenia in men aged over 55 years with T2DM was 34.86%. The BMD of the participants in sarcopenia group was significantly lower than that in non-sarcopenia group. Multivariate Logistic regression analysis showed sarcopenia was an independent risk for osteoporosis in men with T2DM(OR=2.76, 95% CI: 1.20~6.34, P=0.017). Conclusion The sarcopenia is closely related to osteoporosis in middle-aged and aged men with T2DM.

Risk factors and pathogenic bacteria of catheter-related blood stream infection in ICU

Qiao Li, Cao Yang, Yuan Hongxun, Wang Yao

2020, 41(1):  125-130.  doi:10.3969/j.issn.1006-7795.2020.01.024

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Objective To understand the infection rate, risk factors, pathogen species, drug resistance and prognosis of catheter-related blood stream infection (CRBSI) in the Intensive Care Unit (ICU) of Peking University International Hospital, and to provide guidance for reducing the incidence of CRBSI. Methods Clinical and microbiological data of patients diagnosed with CRBSI from January 1, 2016 to December 31, 2018 in the Intensive Care Unit of Peking University International Hospital were retrospectively analyzed. Results This study involved in 674 cases of central venous catheter indwelling, 6 197 days of catheterization, and 28 cases of CRBSI, with an infection rate of 4.52‰. A total of 36 strains of pathogenic bacteria were cultured in 28 patients, including 18 Gram-positive strains (50%).There were 16 Gram-negative bacteria (44.4%) and 2 fungi (5.6%). Staphylococcus epidermidis dominated by G+ bacteria, acinetobacter baumannii dominated by G-bacteria, and candida albicans dominated by fungi. Logistic multiple regression analysis showed that heavy antibiotic use and indwelling catheter duration were independent risk factors for CRBSI in the ICU. Conclusion The incidence of CRBSI in ICU is still high, and the pathogen is mainly G+ bacteria.

Oxidative stress mediates the effect of NLRP3 inflammasome on chronic obstructive pulmonary disease

Liu Ping, Lou Ke, Weng Long, Li Hui, Tang Yuling

2020, 41(1):  131-136.  doi:10.3969/j.issn.1006-7795.2020.01.025

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Objective The correlation among nucleotide binding oligomerization domain like receptor protein 3(NLRP3), malondialdehyde(MDA), superoxide dismutase(SOD) and forced expiratory volume in the first second% pred(FEV1% pred) was analyzed by detecting the levels of SOD, MDA and the expression level of NLRP3 mRNA in serum of COPD patients, so as to explore the influence of oxidative stress-mediated NLRP3 inflammatory body activation on COPD. Methods Selection of Changsha's Hospitals affiliated to Nanhua University in June 2017 to June 2018, 90 cases of COPD patients including 60 patients with acute chronic obstructive pulmonary disease (AECOPD).According to pulmonary function in patients with AECOPD group is divided into weight very restructuring and light-medium group, 30 cases in each group.Selected 30 COPD patients who reached the level remission after treatment symptoms and selected the other 30 cases who got healthy subjects through physical examination as a control group.Serum MDA, SOD levels and NLRP3 mRNA expression levels in AECOPD group, COPD stable period group, control group were compared. Otherwise, serum MDA, SOD levels and NLRP3 mRNA expression levels in weight-very restructuring group, light-medium group and healthy control group were compared.Pearson correlation analysis was used to analyze the correlation between serum MDA and SOD levels, NLRP3 mRNA expression in peripheral blood lymphocytes, and FEV1% pred in COPD patients.Results Serum MDA level in AECOPD group was higher than that in COPD stable period group and COPD stable period group was higher than that in healthy control group (P<0.05). Serum SOD level in AECOPD group was lower than that in COPD stable period group and COPD stable period group was lower than that in healthy control group (P<0.05).The expression level of NLRP3 mRNA in peripheral blood lymphocytes in AECOPD group was higher than that in COPD stable period group and COPD stable period group was higher than that in healthy control group (P<0.05). Serum MDA level in the weight-very restructuring group was higher than that in the light-medium group and the light-medium group was higher than that in the healthy control group (P<0.05). Serum MDA level in the weight-very restructuring group was lower than that in light-medium group and light-medium group was lower than that in the healthy control group (P<0.05). The expression level of NLRP3 mRNA in peripheral blood lymphocytes in weight-very restructuring group was significantly higher than that in the light-medium group and the light-medium group was higher than that in the healthy control group (P<0.05). The result of pearson correlation analysis showed that serum MDA level in COPD patients was positively correlated with NLRP3 mRNA expression, with a correlation coefficient r of 0.64(P<0.05). Serum SOD level was negatively correlated with NLRP3 mRNA expression in COPD patients, with a correlation coefficient r of -0.90(P<0.05). In COPD patients, FEV1%pred was negatively correlated with NLRP3 mRNA expression, and the correlation coefficient r was -0.78 (P<0.05). Conclusion Oxidative stress and NLRP3 inflammatory corpuscle are involved in the pathogenesis of COPD. The increasing of oxidative stress and NLRP3 mRNA expression were correlated with exacerbations of COPD. Oxidative stress-mediated activation of NLRP3 inflammatory corpuscle has an impact on the progression of COPD.