1.Construction of shRNA eukaryotic expression vector targeting OPTN gene and detection of interference effect
Yu GU ; Libin FAN ; Xiaoyin LIU
Acta Universitatis Medicinalis Anhui 2014;(1):1-4
Objective To construct shRNA against optineurin ( OPTN ) clone and transfect the construction into HEK293FT cells for inhibiting the expression of OPTN, which is laid the foundation for further research of molecu-lar mechanism OPTN protein. Methods The insert of this clone was a double-strained DNA sequence against OPTN which would be transcripted into shRNA and it was synthesized according to the sequence in Origene. Ex-pression vector pRFP-C-RS was employed to fuse the insert to get the construction and finally confirmed by sequen-cing. HEK293FT cells were transfected with the very clone pRFP-C-RS-shOPTN and RFP signals could be detected using fluorescence microscope. Western blot was further employed to check the protein level of OPTN. HEK293FT cells were transfected with the very clone pRFP-C-RS-shOPTN was then obtained to investigate the functional study of OPTN which was measured by Salmonella infection experiment. Results shRNA eukaryotic expression vector targeting OPTN gene was successfully constructed. HEK293FT cells were transfected with pRFP-C-RS-shOPTN and 72 hours later we observed strong RFP signals showing that shOPTN expressed. Western blot was further employed to check the protein level of OPTN which showed that OPTN expression was indeed interfered. Salmonella infection assays was then performed and showed that OPTN could inhibit the proliferation of Salmonella that has invaded HEK293FT cells. Conclusion HEK293FT cell line which expresses shRNA against OPTN show a sharp inhibition of OPTN protein expression by 80%. So this cell line can be further used to investigate how OPTN regulates auto-phagy as an autophagy receptor. Meanwhile we find that OPTN can be as a autophagy regulator and strongly sup-press the proliferation of invasive Salmonella in vivo.
2.Effects of morphine preconditioning on myocardial ischemia-reperfusion injury and phosphorylated extracellular signal-regulated kinase 1/2 expression in rats wRh chronic heart failure
Yunxiang WU ; Ye ZHANG ; Fan JIANG ; Lijun WENG ; Xianwen HU ; Yun LI ; Libin FAN
Chinese Journal of Anesthesiology 2011;31(7):854-857
ObjectiveTo investigate the effects of morphine preconditioning on myocardial ischemiareperfusion (I/R) injury and the expression of phosphorylated extracellular signal-regulated kinase 1/2 (p-ERK1/2)in rats with chronic heart failure.MethodsForty-eight healthy male SD rats weighing 220-250 g were randomly divided into 6 groups ( n =8 each):control group (group C),sham operation group (group S),I/R group and preconditioning with low,median and high doses of morphine groups (groups MP1-3 ).Chronic heart failure was induced by iv edriamycin 2.0 mg/kg once a week for 6 weeks in groups S,I/R and MP1-3.Left ventricular end-diastolic diameter (LVEDD) and left ventricular end-systolic diameter (LVESD) were measured using ultrasound,and left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (LVFS) were calculated at the end of 14th day after the end of adriamycin administration.Blood samples from the carotid artery were collected after ultrasonography for determination of the plasma brain natriuretic peptide (BNP) concentration.Myocardial I/R was induced by 30 min occlusion of left anterior descending branch of coronary artery followed by 120 min reperfusion at 2 day after ultrasonography in groups I/R and MP1-3.In groups MP1-3,iv morphine 0.015,0.030 and 0.050 mg/kg were repeated 3 times at 5 min interval at 30 min before ischemia respectively,while normal saline 5 ml/kg was given in group I/R.The animals were sacrificed at the end of reperfusion in groups S,I/R and MP1-3,and the hearts were removed to measure the area at risk (AAR),infarct size (IS),and IS/AAR ratio was calculated.The p-ERK1/2 expression in myocardium was assessed by Western blot.ResultsThe LVESD and plasma BNP concentration were significantly higher,while the LVEF and LVFS lower in the other 5 groups than in group C (P <0.01).No myocardial infarction was found in group S.The p-ERK1/2 expression was significantly lower in groups I/R and MP1 than in group S (P < 0.05).IS and IS/AAR ratio were significantly lower,and p-ERK1/2expression was significantly higher in groups MP2.3 than in group I/R ( P < 0.05).There were no significant differences in IS,IS/AAR ratio and p-ERK1/2 expression between groups MP1 and I/R (P > 0.05).IS and IS/AAR ratio were decreased gradually,and the p-ERK1/2 expression was up-regulated gradually in groups MP1-3 ( P <0.05).ConclusionMorphine preconditioning can confer cardioprotection against myocardial I/R in a dose-dependent manner in rats with chronic heart failure.Up-regulation of p-ERK1/2 expression is involved in the underlying mechamism.
3.Study on reverse transsynaptic virus tracing targeting Shenmen (HT7) and heart in mice
Libin WU ; Fan ZHANG ; Qing YU ; Lei LIU ; Xia WEI ; Ronglin CAI ; Zijian WU ; Ling HU
Journal of Acupuncture and Tuina Science 2023;21(2):109-120
Objective: To investigate the neural connections between Shenmen (HT7)-heart and the brain by observing the tracing viruses co-labeled brain nuclear groups after injection of the pseudorabies viruses (PRV), the reverse transsynaptic virus tracer carrying different fluorescent protein genes, into the myocardium and Shenmen (HT7) point, respectively.Methods: Pseudorabies virus 531 (PRV531) carrying the green fluorescent protein gene and pseudorabies virus 724 (PRV724) carrying the red fluorescent protein gene were injected into the left ventricular wall and Shenmen (HT7) point area of the left forelimb of six C57BL/6 mice, respectively. After 120 h, whole brain tissue was extracted under 4% paraformaldehyde perfusion to prepare brain sections. Neuronal co-labeling with the tracing viruses was observed under fluorescence microscopy. Results: Co-labeled signals from the mouse ventricular wall and Shenmen (HT7) point region were found at all levels of the mouse central nervous areas, such as the cerebral cortex, hypothalamus, midbrain, pons, and medulla oblongata. The number of co-labeled neurons was higher in the primary motor area, the hypothalamic paraventricular nucleus, the subceruleus nucleus, and the paramedian reticular nucleus. Conclusion: There is a neural connection between Shenmen (HT7), the heart, and the brain, which may be most closely related to the autonomic nervous system.
4.Protective effects of Dahuangfuzi decoction on the intestine barrier functional of acute necrotizing pancreatitis in rats
Xiaoguang LU ; Libin ZHAN ; Xin KANG ; Guohui LIU ; Yun DONG ; Zhiwei FAN ; Lizhi BAI ; Li LIU ; Chunyang JI ; Xiaozhou WANG
Chinese Journal of Pancreatology 2011;11(2):113-116
Objective To observe the effects of Dahuangfuzi decoction on the intestine barrier functional of acute necrotizing pancreatitis in rats. Methods The 60 rats were randomly divided into sham operation group ( n = 19 ), ANP group ( n = 21 ), and Dahuangfuzi treatment group ( n = 20). The rats of ANP group were induced by injecting 1 ml/kg of 4% sodium taurocholate into the pancreatiobiliary duct, and jejunal fistula was esablished. The rats of treatment group received Dahuangfuzi decoction (2 ml, repeated at 4 and 8 h)through jejunum distal stoma tube 0. 5 h after ANP induction. The other 2 groups received same amount of normal saline. Blood sample was collected through abdominal aorta, 24 h after ANP induction, and the serum amylase, endotoxin, D-lactate, plasma diamine oxidase (DAO) were detected. Pancreas, small intestine tissue was harvested for pathologic examination, index of intestinal epithelial damage was measured and ultrastructural changes in small intestinal mucosa was observed. Results The expression of serum amylase, endotoxin,D-lactate, DAO in sham operation group was ( 152 ± 32 ) U/L, (6.95 ± 2.10) pg/L, ( 3.96 ± 1.08 ) μg/mland ( 14.26 ± 2.67 ) μg/ml, while the corresponding values were ( 1549 ± 93 ) U/L, (40.48 ± 3.41 ) pg/L,( 12.34 ± 1.23 ) μg/ml and ( 80.28 ± 3.54) μg/ml in ANP group, and they were (655 ± 49 ) U/L, ( 19.55 ±2.50) pg/L, (6.75 ± 1.36 ) μg/mland ( 20.69 ± 7.53 ) μg/ml in treatment group. The values in ANP group were significantly higher than those in sham operation group. The values in treatment group were significantly lower than those in ANP group, but significantly higher than those in sham operation group ( P < 0.05 or P <0. 01 ). The thickness and height of intestinal mucosa in ANP group were ( 389.44 ± 29.87 )μm and ( 16.52 ±3.73) μm, which were significantly lower than those in treatment group [(501.95 ± 45.38 )μm, (27.82 ±5.17)] μm, and in sham operation group [( 658.72 ± 57.49 ) μm, ( 35.49 ± 6.43 )μm, Index of intestional epitholial donage in ANP group was 3.72 ± 0.65 which is significently higher than those in theatment (2.12 ±0.37 ) and in sham operation group (0.85 ± 0.24). The intestinal mucosa histological and ultrastructural changes in Dahuangfuzi treatment group were better than those in ANP group. Conclusions Dahuangfuzidecoction can significantly decrease the damage of intestine barrier function in ANP rats.
5.Development of evaluation index system of reputation for higher medical institutions based on fac-tor analysis method
Tao SUN ; Depin CAO ; Libin YANG ; Linli YANG ; Wei ZHANG ; Siao GE ; Lihua FAN ; Li LI ; Xinyan LIU ; Lei GAO
Chinese Journal of Medical Education Research 2016;15(2):109-113
Objective To develop an evaluation index system of reputation for higher medical insti-tutions and to test its reliability and validity. Methods Using literature method and small-group discussion to collect the evaluation indexes, the data was collected by the questionnaire survey and the 1 493 effective questionnaires were collected with effective recovery rates of 61.90%. And the exploratory and confirmatory factor analysis methods were conducted to develop evaluation index system with using the SPSS 17.0 soft-ware for statistics analysis. Results The evaluation index system of reputation for higher medical institutions included six dimensionalities: social responsibility, opinion direction, social image, management quality, emotional attract and environmental facility. The Cronbachs’a coefficients of evaluation index system were greater than 0.7, RMSEA is 0.063<0.08, χ2/df is 3.933<5, therefore, the structural equation model fitting was good and evaluation index system had good validity. Conclusion This article has developed an evalu-ation index system of reputation for higher medical institutions, which has a good reliability and validity. To test the model’s reliability and validity, the suggestion that the future research will be performed between more other medical universities has been given.
6.In vivo biocompatibility of nano-hydroxyapatite/polyphenylene sulfide composites
Kexia FAN ; Yuan MA ; Weizhong YANG ; Sixun YU ; Zhiyong SUN ; Xun XIA ; Libin YANG ; Heng GUO ; Yongqin KUANG ; Jianwen GU
Chinese Journal of Tissue Engineering Research 2017;21(22):3547-3554
BACKGROUND:There is a lack of the research concerning the biocompatibility of nano-hydroxyapatite/polyphenylene sulfide (nHA/PPS) composites.OBJECTIVE:To evaluate the in vivo biocompatibility of nHA/PPS composites based on the completed research in vitro.METHODS:Systemic toxicity test:Sprague-Dawley rats were given the intraperitoneal injection of nHA/PPS extract or normal saline.The general situation,body mass and the histological changes of the liver and kidney were observed at 72 hours after injection.Delayed type hypersensitivity test:nHA/PPS extract or normal saline was injected subcutaneously into the back of the rats.Afterwards,skin irritation symptoms were observed at 72 hours.Local reaction experiment:nHA/PPS composites and polyethylene were respectively implanted into the back of the rats.The pathological changes of the implanted materials and their surrounding tissues were observed at 15 and 30 days after implantation.RESULTS AND CONCLUSION:(1) The rats were in good situation after nHA/PPS injection;the body mass increased steadily,which showed no significant difference from the control group (P < 0.05);the morphology and color of the liver and kidney were normal,and the systemic toxicity of the composite materials was normal according to the degree of toxicity classification.(2) There were no obvious skin irritation symptoms after the subcutaneous injection of nHA/PPS composites,and the primary irritation index was less than 0.4,suggesting a low hypersensitivity.After implantation of nHA/PPS composites,there was no obvious degradation,absorption and rejection,and both the degree of inflammatory reaction (15 days ≤ level Ⅲ,30 days ≤ level Ⅱ) and the thickness of fibrous capsule (15 days ≤ level Ⅲ,30 days ≤ level Ⅱ) revealed the good biocompatibility of the composites.These results suggest that the nHA/PPS composites hold an excellent biocompatibility in vivo.
7.Application of independent component analysis for separating phonocardiogram signals.
Xiumei YANG ; Jiahua PAN ; Zuxing ZHANG ; Libin FANG ; Yuanxia JI ; Yun FAN ; Weilian WANG
Journal of Biomedical Engineering 2008;25(4):766-769
Independent component analysis (ICA) is a novel method developed in recent years for Blind Source Separation. In this paper, the phonocardiogram (PCG) was separated into three components by applying ICA. The basic principle of ICA was introduced in this paper. A fast and robust fixed-point algorithm for ICA was used to analyze PCG signals in this study. The experiments showed that ICA could separate the components of heart sounds from PCG signals successfully.
Algorithms
;
Heart Sounds
;
Humans
;
Phonocardiography
;
methods
;
Principal Component Analysis
;
Signal Processing, Computer-Assisted
8.Effect of Dahuang Fuzi decoction on alveolaur epithelial barrier in rats with lung injury with severe acute pancreatitis
Xiaoguang LU ; Xin KANG ; Libin ZHAN ; Dianbo GONG ; Li LIU ; Zhiwei FAN ; Lizhi BAI ; Honggang PANG ; Limin KANG ; Chunyang JI ; Xiaozhou WANG
Chinese Journal of Emergency Medicine 2011;20(2):151-155
Objective To investigate effect of Dahuang Fuzi decoction on alveolaur epithelial barrier in rats with lung injury with severe acute pancreatitis. Method Ninty-six health SD rats were randomly divided into three groups: sham operation group, SAP-ALI group, Dahuang Fuzi decoction group, and then according to the time point of sacrifice after operation, each group was subdivided into 3,6,12,24 hour subsets ( each, n = 8). After the belly of a rat in the sham operation group was cut open, the pancreas was flipped several times,and then a stoma was made in the jejunum to form its fistula. In the SAP-ALl group,1 mL/kg sodium taurocholate was reversely injected into the pancreatobile duct to establish the model of SAP, and then the jejimum fistula was performed. The SAP-ALI model in Dahuang Fuzi decoction group was treated by injection of 10ml of Dahuang Fuzi decoctionon into the fistula respectively. Blood was collected from heart to detect serum amvlase and endotoxin (ET) levels before the rat being executed. The lung histopathologic changes, pulmonary injury scores and wet/dry weight(W/D) ratios were observed after the rats were executed. The alveolar liquid clearance rate(ALCR), total lung water content (TLW), extravascular lung water content(EVLW) and alveolar epithelial permeability (AEP) were examined in 3,6, 12,24 h after injury.Results There was continuous increase of AEP,TLW and EVLW,as well as progressive reduction of ALCR compared with sham operation group at 3,6,12,24 h after operation. Compared with SAP-ALI group, there was continuous decrease of AEP,TLW and EVLW, and elevated of ALCR at 3,6,12,24 h after operation.Conclusions Dahuang Fuzi decoction can significantly reduce alveolaur epithelial barrier and degree of lung tissue of SAP-ALI rats by inhibiting the elevation of LPS and inflammation reaction.
9. A bibliometric analysis of literature on hand-transmitted vibration in China, 1990-2016
Liuquan JIANG ; Gaisheng LIU ; Zhizhong YANG ; Xiaojun SHEN ; Fan YANG ; Zhechen ZHANG ; Libin GONG
Chinese Journal of Industrial Hygiene and Occupational Diseases 2017;35(12):932-935
Objective:
To investigate the features of literature on hand-transmitted vibration in China, 1990-2016.
Methods:
In September 2017, the studies on hand-transmitted vibration in China, which were published in Chinese or English during 1990-2016, with "China" and "Taiwan" as the places where author affiliations were located, were retrieved. A bibliometric analysis was performed to investigate the type of articles, publication time, the journals in which articles were published, author affiliations, author regions, and funding.
Results:
A total of 205 articles on hand-transmitted vibration were retrieved. There were 7.59 articles on average published annually from 1990 to 2016. In the 205 articles, 114 (55.61%) were published in the journals indexed in one or two core journal databases. In the 64 journals, 22 (34.38%) were indexed in one or two core journal databases. The first authors were from 22 provincial regions (provinces, autonomous regions, or centrally administered municipalities) in China, with 152 articles (74.15%) by the authors in the top five regions. There were a total of 876 authors, and the co-authorship degree was 4.27 (876/205). Most of the first authors (136 articles, 66.34%) were affiliated with universities or institutes for prevention and control of occupational diseases. Among the 205 articles, 103 (50.24%) were original articles or investigations, and 72 (35.12%) were funded.
Conclusion
The studies on hand-transmitted vibration fluctuated and increased from 1990 to 2016, with a relatively concentrated distribution in terms of sources, regions, and institutions. Interregional and international academic exchange should be strengthened.
10.Influencing factors for hierarchical medical system:interpretative structural modeling
Dexin MENG ; Shu′e ZHANG ; Chao FAN ; Yifu RU ; Haiyan LI ; Yu SHI ; Fengzhe XIE ; Libin YANG ; Mohan CHI ; Dachuan MAO ; Tao SUN
Chinese Journal of Hospital Administration 2016;32(7):481-484
Objective To build a multi‐level hierarchical structure model of the influencing factors for hierarchical medical system ,to identify the role relationship between all the factors and transmission pathways ,and to recommend on developing China′s hierarchical medical system . Methods Thirty influencing factors were identified in a screening based on literature review for the hierarchical medical system .On such basis ,16 influencing factors were identified by three health policy experts ,which affect operations of the current system .Interpretative structural modeling was called into play in the end to analyze the hierarchy relationship between various influencing factors and the conduction loops .Results There exist among the 16 factors a 3‐level hierarchical structureand two conduction loops .The factor directly limiting the hierarchical medical system is two‐way referral,and most internal core drivers arehuman resources development and governance mechanism.By means of self‐growth and external constraints ,they exert their influence on the operation of hierarchical medical system .Conclusions There are interactive hierarchical effects among the factors ,merging into three node elements of functional role,inter‐entity relationshipand patient participation.