1.Effects of hyperbaric oxygen on the intestinal mucosal apoptosis caused by ischemia-reperfusion injury in rats
Shihui ZHOU ; Yanfei SUN ; Gang WANG
Chinese Journal of Emergency Medicine 2010;19(12):1281-1286
Objective To observe the effects of hyperbaric oxygen (HBO) on the apoptosis expression of intestinal mucosa during the different periods of ischemia-reperfusion injury in order to elucidate the underlying mechanisms. Method Rats were subjected to ischemia for 60 min by clamping superior mesenteric artery, and then had reperfusion for 60 min by unclamping. Rats were randomly divided into four groups: ischemia-reperfusion group (I/R), pre-emptive HBO or HBO treatment before ischemia (HBO-P) group, HBO treatment during ischemia period (HBO-I) group, and HBO treatment during reperfusion (HBO-R) group. After reperfusion for 60 min, samples of small intestine tissue were taken from the end portion of ileum for detecting the levels of ATP by using colorimetric method and the levels of caspase-3 by using immunochemistry. The levels of TNF-α in intestinal tissue were measured by using enzyme-linked immunosorbent assay method ( ELISA). All values were expressed in Mean ± Standard Deviation (x ± s). The different groups were compared among them with SNK- q test of OneWay analysis of variance (One-Way ANOVA plus SNK). Results The levels of TNF-α in HBO-I group were significantly lower than that in HBO-P group ( P < 0.05), and significantly lower in HBO-P group than those in HBO-R or I/R groups ( P < 0.05), and there was no significant difference in TNF-α between HBO-R and I/R group ( P > 0.05). The levels of caspase-3 were significantly lower in HBO-I group than those in HBO-P group ( P < 0. 05), and also significantly lower in HBO-P group than those in I/R or HBO-R groups ( P < O. 05), and no significant difference caspase-3 was found between HBO-R and I/R groups. The ATP levels were significantly lower in HBO-I group than those in HBO- P group ( P < 0. 05), and also significantly lower in HBO- P group than those in I/R or HBO-R group ( P < 0.05), and no significant difference in ATP level between in HBO-R and I/R group. Conclusions There was a connection between HBO and small intestinal I/R injury as well as mucosal cell apoptosis. And HBO maintained ATP and aerobic metabolism, and inhibited the genesis of TNF-α, and thus in turn prevented intestinal mucosa cell from apoptosis. The best result was obtained when HBO was administered during ischemia period, and there was no effect found when HBO was employed during reperfusion period.
2.Color damage in patients with idiopathic optical neuritis after the treatment
Tangsheng SUN ; Mujiang AXI ; Shihui WEI
Chinese Journal of Ocular Fundus Diseases 2010;26(4):335-338
Objective To detect the color damage in patients with idiopathic optical neuritis (ION) after the treatment. Methods A total of 26 ION patients (44 eyes) with ION whose visual acuity were above 1. 0 were collected. All the patients had undergone the treatment of incretion and had the visual acuity more than 1. 0 after the treatment. The results of MRI and blood examination were normal. Another 24 healthy persons were selected as the normal control. Total error scores (TES) and each error score of red, green and blue were measured via Farnsworth Munsell-100 hue tester. The TES origin scores and their square roots were used for a statistical analysis. The results of the two groups were compared. Results There were significant differences in TES and its square roots between ION group and the normal control group (t=3.079,3. 133;P = 0. 0033,0. 0026). The differences in the level of error scores of each color between the tow groups were not significant (t = 1. 91, 1.15, 1.62;P = 0.061, 0.26, 0.11);but the differences in the square roots of red color between the two groups were statistically (t=2. 21, P = 0. 031).Conclusion After the treatment, the visual acuity of ION patients increases, but the color damage still exist;red color damage happens first.
3.Analysis on the Traditional Chinese Medicine Syndromes of 177 HIV/AIDS P atients
Miaomiao ZHANG ; Linchun FU ; Shihui SUN
Journal of Traditional Chinese Medicine 1993;0(04):-
Objective To study the rule of distribution and evol vement of TCM syn dromes in HIV/AIDS patients. Methods Totally 177 HIV/AI DS patients were investigated with questionnaire in the epide miological study to collect the data of diagnosis, and the rou tine test of CD4+T lymphocyte count was carried out. Results The young and mid dle-aged females were the most involved group with an increasing incidence in G u angdong Province and with a strong relation of sex and infection route. The main symptoms of the 177 HIV/AIDS patients were fatigue, cough and loss of appetite, the secondary ones were night sweating, skin itching, headache, and hair loss, and the most common sign was skin rash. The leading TCM syndromes were qi and yi n deficiency and lung and kidney deficiency. The distribution of symptoms and si gns, syndromes and various indices was significantly different in gender, age an d infection route (P
4.Different dose and effects of dexmedetomidine in preventing shivering after general anesthesia for laparoscopic surgery
Yijuan SUN ; Hui LUO ; Xizhao HUANG ; Shihui YANG ; Zurong HU
The Journal of Practical Medicine 2014;(12):1976-1978
Objective The aim of this study was to investigate different dosages and effects of dexmedetomidine for prevention of postanesthetic shivering. Methods One-hundred twenty patients scheduled for laparoscopic surgery were randomly allocated in four groups: before the operation, slowly injected 0.9% normal saline (group S, dexmedetomidine 0.5 μg/kg (group D0.5), dexmedetomidine 0.75 μg/kg (group D0.75), dexmedetomidine 1.0 μg/kg(group D1.0). HR and rectal temperature[C2] were continually monitered during and after operation, time to extubation was measured. Grades of shivering were recorded. Pain evaluation was assessed by a visual analogue scale, sedation was evaluated by Modified Observer′s Assessment of Alertness/Sedation scale. Results The patients in group S showed a significantly higher HR and postoperative incidence of shivering than those in group D0.75 and group D1.0, (P < 0.05). but the extubation time in groupd D0.75 and group D1.0 were longer than patients in group S (P<0.05). Conclusion Slowly injected dexmedetomidine 0.75 μg/kg or 1.0 μg/kg can prevent postanesthetic shivering in laparoscopic surgery effectively.
5.A systematic review of the effectiveness of glucocorticoids in the treatment of nonarteritic anterior ischemic optic neuropathy
Wanyu SUN ; Xing LIAO ; Ying WANG ; Shihui WEI
Chinese Journal of Ocular Fundus Diseases 2021;37(3):224-230
Objective:To evaluate the effectiveness and safety of glucocorticoids in the treatment of non-arteritic anterior ischaemic optic neuropathy (NAION).Methods:Glucocorticoids published in the National Library of Medicine PubMed; Netherlands Medical Abstracts Database Embase; Cochrane Library, an evidence-based medical library; China Cnkipedia; China Biomedical Literature Service; Chongqing Vipul Chinese Science and Technology Journal Database, and Wanfang Science and Technology Journal Full Text Database were searched about computer. Randomized controlled trials (RCTs) and non-randomized controlled trials (NRCTs) for the treatments of NAION were subjected to meta-analysis. The search period was from the establishment of each database to March 2020. The literature was screened and data were extracted according to the inclusion and exclusion criteria. The methodological quality of the RCT and NRCT studies was evaluated using the Risk of Bias Assessment Tool and the MINORS evaluation scale, respectively. The data were analyzed using RevMan version 5.3 software which was provided by the Cochrane Collaboration Network.Results:An initial search of 395 papers was conducted, and 10 papers were finally included for this meta-analysis, including 3 RCT studies and 7 NRCT studies. A total of 1057 patients with NAION were included. The 3 RCT studies were analyzed descriptively as the outcome indicators were described in different ways. A meta-analysis of 7 NRCT studies showed that patients in the treatment group showed significantly better visual prognosis (relative risk=1.28, 95% confidence interval 1.09 to 1.51, P=0.003) and retinal nerve fibre layer thickness were obviously improved (mean difference=7.76, 95% confidence interval 1.58 to 13.94, P=0.01) than the control group. Four studies reported the occurrence of adverse reactions in the treatment versus control groups. None of the above studies provided a detailed analysis of the prognosis of patients with adverse reactions. Conclusion:The efficacy and safety of glucocorticoids in the treatment of NAION is unclear and needs to be validated in a larger sample of RCTs.
6.Establishment of a mouse model of acute liver failure induced by LPS/D-GalN
Xiaohong WU ; Yan GUO ; Chenfeng LIU ; Tongtong GAO ; Hong YU ; Shihui SUN ; Yusen ZHOU
Acta Laboratorium Animalis Scientia Sinica 2014;(3):15-19
Objective To establish a mouse model of acute liver failure induced by lipopolysaccharide /D-galac-tosamine ( LPS/D-GalN) .Methods The optimum dose of LPS/D-GalN was determined by i .p.injection of eight differ-ent doses of LPS and D-GalN into 40 female C57BL/6 mice and observation of their survival time .Then, 32 female C57BL/6 mice were i.p.injected with the optimal dose of LPS/D-GalN and sacrificed at 0, 1, 4, 8 hours after the injec-tion, 8 mice in each group.The control mice received saline injection .Hepatic changes were observed by pathology and se-rum ALT, IL-6, MCP-1 and TNF-αwere measured by biochemistry or flow cytometry .Results LPS (2.5 mg/kg) and D-GalN (0.3 g/kg) were determined as the optimal dose for the establishment of mouse model of acute liver injury .Com-pared with the control group , the hepatocellular damages were progressing in a positive correlation with the time course after LPS/D-GalN administration .The level of serum ALT was significantly increased after LPS/D-GalN administration ( P <0.001).The levels of inflammatory cytokines IL-6, MCP-1 and TNF-αwere increased and reached a peak at one hour after LPS/D-GalN administration and then decreased almost to that of the control group 8 hours later(P<0.001).Conclusions The mouse model of acute liver injury is successfully established by LPS /D-GalN administration , and provide an effective animal model for the study of pathogenic mechanisms of acute liver failure and evaluation of therapeutic drugs .
7.Stable expression of targeting complement inhibitor CR2-CD59 in Chinese hamster ovary cells
Yan GUO ; Zhihua KOU ; Shihui SUN ; Chuanfu ZHANG ; Guangyu ZHAO ; Hong YU ; Hongbin SONG ; Fei QIAO ; Wanrong CHEN ; Yusen ZHOU
Journal of Third Military Medical University 2003;0(09):-
Objective To obtain Chinese hamster ovary (CHO) cell lines that stably express a targeting complement inhibitor CR2-CD59.Methods The recombinant plasmid PEE14.1-CR2-CD59 was constru-cted by cloning the DNA fragment CR2-CD59 into plasmid PEE14.1,and the obtained plasmid was transfected into CHO cells by FuGENE 6.The clones with stable high expression of target fragment were selected by methionine sulfoximine (MSX),the expression of CR2-CD59 was analyzed by ELISA,SDS-PAGE and Western blotting analysis.Results Several stable expression clones were obtained,and CR2-CD59 was highly expressed in the secret form in CHO cells.SDS-PAGE analysis showed that the molecular weight of the recombined protein CR2-CD59 was consistent with the predicted one.ELISA and Western blotting results revealed that the CR2-CD59 could react with both anti-human CR2 and anti-human CD59 polyclonal antibodies.Compared with serum-containing medium,the protein was highly expressed in serum-free medium (P
8.Analysis of clinical features and prognostic factors of aquaporin 4 antibody positive neuromyelitis optica spectrum disorders related optic neuritis
Xintong XU ; Mo YANG ; Huanfen ZHOU ; Mingming SUN ; Shihui WEI
Chinese Journal of Ocular Fundus Diseases 2022;38(12):968-973
Objective:To analyze the clinical features and prognosis factors of aquaporin 4 (AQP4) antibody-positive neuromyelitis optica spectrum disorders related optic neuritis (NMOSD-ON).Methods:An ambidirectional cohort study. From June 1, 2015 to June 1, 2019, 103 patients with AQP4 antibody-positive NMOSD-ON in Department of Neuro-ophthalmology, The First Medical Center of PLA General Hospital were included. All patients of followed-up period were ≥24 months. According to the best corrected visual acuity (BCVA) at the last follow-up, the affected eyes were divided into the low vision group [log of minimum resolution angle (logMAR) BCVA≥1.0] and the non-low vision group (logMAR BCVA<1.0), 66 and 37 cases, respectively. The two groups of patients were compared the genernal clinical characteristics, and the logistic regression model and COX proportional hazard model were used to analyze the relevant factors affecting the patient's visual prognosis and recurrence.Results:Among the 103 cases, 96 cases (93.2%, 96/103) were female; 94 cases (91.3%, 94/103) had unilateral disease; 48 cases (46.6%, 48/103) were the first onset; 85 cases (82.5%, 85/103) were effected by eye pain or orbital pain; 21 cases (20.4%, 21/103) had optic disc edema; 51 cases (49.5%, 51/103) serologically autoimmune antibody test were positive. Orbital magnetic resonance imaging (MRI) was performed in 101 cases. There was no obvious abnormal signal in visual pathways except for 5 cases (5.0%, 5/101); 96 cases (95.0%, 96/101) had abnormal signal in the visual path, and the optic nerve was found in the orbit; 52 cases had abnormal optic nerve in orbital segment (51.5%, 52/101); 37 cases (35.9%, 37/103) recurred within 24 months. The recovery of logMAR BCVA after the first onset and the logMAR BCVA at the first onset, at 6 months of follow-up in two groups were 1.4±1.0, 0.3±0.4, 1.9±0.7 and 0.4±0.5, 2.1±0.6, 0.3±0.4, respectively; and there were statistically significant differences between the two groups of patients at different times ( Z=-4.967,-7.603,-8.027; P<0.001). Logistic regression multivariate analysis showed that recovery of BCVA≥1.0 logMAR after the first onset [odds ratio ( OR)=226.276, P<0.001 ] and the number of attacks ( OR=8.554, P=0.003) were independent risk factors for low vision. Multivariate analysis of the Cox proportional hazards model showed the higher the MRI score [hazard ratio ( HR)=0.588, P=0.007] and plasma exchange ( HR=0.124, P=0.049) in the acute phase were protective factors for recurrence. Conclusions:Vision loss accompanied by eye pain or orbital pain is the main symptom of onset AQP4 antibody-positive NMOSD-ON, a small number of patients have disc edema, 49.5% patients serologically autoimmune antibody test are positive. Abnormal optic nerve signals can be seen in 95.0% of patients in orbital MRI, and 51.5% patients have abnormalities in the orbital optic nerve. The worse the recovery of BCVA after the first onset and the greater the number of attacks are unfavorable factors affecting the prognosis of vision. High MRI scores and plasma exchange in the acute phase are favorable factors to prevent the recurrence of the disease.
9.Construction and validation of a dual-luciferase reporter gene system for screening and evaluating anti-liver fibrosis drugs that inhibit transcription of the gene encoding collagen I, chain a1.
Wei ZHANG ; Xiaoming DAI ; Hong YU ; Luyan WANG ; Shihui SUN ; Junfeng LI ; Yusen ZHOU
Chinese Journal of Hepatology 2014;22(10):747-751
OBJECTIVETo construct a dual-reporter gene system that will be applicable for use as a tool to screen and evaluate therapeutic drug compounds that inhibit transcription of the gene encoding collagen I, chain at1 (COL1A1).
METHODSThe full-length eDNA of transforming growth factor beta1 (TGFbeta1) was cloned by RT-PCR and inserted into two vectors, pcDNA3.1 and pJW4303, for construction of two eukaryotic expression vectors, pcDNA3.1-TGFbeta1 and pJW4303-TGFbeta1.Next, the promoter region of COL1A1, cloned by PCR using human genome DNA as template, was inserted into the vector pGL4.29 to construct the reporter gene vector, pGL4.29-COL1A1 promoter.All three recombinant vectors were verified by restriction enzyme digestion and DNA sequencing.Either the pcDNA3.1-TGFbeta1 or pJW4303-TGFbeta1 vector along with the pGL4.29-COL1A1 promoter vector or a pRL-null, control reporter, vector were co-transfected into the LX-2 human hepatic stellate cells to establish the transcription-activated dualreporter gene system.This system was used as a cell model for screening anti-liver fibrosis compounds that inhibit the transcription of COL1A1.Dexamethasone, a model drug that is known to inhibit the expression of COL1A1, was used as a control to validate the dual-reporter gene system.
RESULTSThe two TGFbeta1-expressing vectors and the reporter gene vector containing the promoter region of COL1A1 were successfully constructed.The results of a dual-reporter gene assay showed that TGFbeta1 co-expression increased the activity of the COL1A1 promoter by above 200-fold (t =21.78, P =0.0001), whereas in the absence of TGF31 co-expression the activity was below 2-fold (t =3.396, P =0.0274).The transcriptionactivated dual-reporter gene system was successfully established.The model drug, dexamethasone, effectively inhibited the activity of the COL 1A1 promoter in dose-dependent manner; the activity decreased 29.6% with 10 mumol/L dexamethasone (t =4.140, P =0.0144) and 53.9% with 100 mumol/L (t =6.193, P =0.0035).
CONCLUSIONThe dual-luciferase reporter system of TGFbeta1 and COL1A1 co-expression developed here can be used as a cell model to screen and evaluate anti-liver fibrosis compounds that inhibit activity of the COL1A1.
Base Sequence ; Collagen Type I ; genetics ; Drug Evaluation, Preclinical ; Genes, Reporter ; Genetic Vectors ; Humans ; Liver Cirrhosis ; drug therapy ; Luciferases ; Promoter Regions, Genetic ; Transcriptional Activation ; drug effects ; Transfection ; Transforming Growth Factor beta1
10.Comparison of pulmonary pathological changes in mice infected with H7N9 influenza virus and pandemic H1N1 influenza virus
Shihui SUN ; Xiaohong WU ; Chenfeng LIU ; Tongtong GAO ; Yang ZENG ; Yan GUO ; Jian TANG ; Ting PAN ; Hong YU ; Zhihua KOU ; Guangyu ZHAO ; Yusen ZHOU
Acta Laboratorium Animalis Scientia Sinica 2014;(3):1-6
Objective To analyze and compare the pathological changes of lung tissue in mice infected with the novel H7N9 influenza virus and 2009 pandemic H1N1 influenza virus, respectively, and to preliminarily study the mecha-nisms of acute lung injury induced by those virus infection .Methods SPF 6-week old BALB/c mice ( body weight 18-20 g, male∶female=1∶1) (n=3 in each subgroup) were intranasally infected with H7N9 virus and H1N1 virus, respec-tively.The behavior and survival time of mice after virus infection were observed and the survival rates were analyzed .The heart, liver, spleen, lung, kidney, intestines, and brain were collected at indicated time points for histopathological exami-nation using H&E staining .The distribution of virus antigen was detected by immunohistochemistry .The neutrophil infiltra-tion was also observed .The correlation of lung injury with virus replication and host immune responses was analyzed .Re-sults The lung and spleen injury of mice infected with H 7N9 virus was slighter and their survival rate (100%) was high-er than those of mice infected with H1N1 virus.The damages of the lung and spleen in H1N1virus-infected mice were more severe than that in H7N9 virus-infected mice, and all the 10 mice in this group died within 9 days after virus inoculation . The distributions of both the virus antigens were mainly in the bronchial epithelial cells , a few stromal cells and alveolar ep-ithelial cells .The levels of virus replication in the two groups were not significantly different .There were more intense neu-trophil infiltration in the lung and inflammatory response in the H 1N1 virus-infected mice than those in the H7N9 virus-in-fected mice .Conclusions There are some differences of the pathological characteristics and extent of lung injury in the mice infected with H7N9 virus and H1N1 virus, respectively.The virus replication is a precipitating factor but not the deci-sive factor of the lung injury , and there is a close relationship between the host immune responses and acute lung injury .