1.Effect of colquhounia root tablet on expression of tight junction proteins claudin-5 and ZO-1 in renal tissues of rats with acute lung injury
Xigang MA ; Ping SHAO ; Xiaohong WANG
Chinese Journal of Anesthesiology 2016;36(1):101-104
Objective To evaluate the effect of colquhounia root tablet on the expression of tight junction proteins claudin-5 and ZO-1 in the renal tissues of rats with acute lung injury (ALI).Methods Twenty-four healthy male Sprague-Dawley rats,weighing 250-300 g,were randomly divided into 3 groups (n =8 each) using a random number table:control group (group C),ALI group;and colquhounia root tablet group (group H).Oleic acid 0.04 ml/kg was injected into the trail vein to establish the model of ALI.In group H,colquhounia root tablet 600 mg · kg-1 · d-1 was instilled into the stomach of each rat for 10 consecutive days,and oleic acid was injected at 1 h after instillation on 10th day.Blood samples were collected from the abdominal aorta at 6 h after injection of oleic acid for determination of plasma creatine (Cr) and blood urine nitrogen (BUN) concentrations.The animals were then sacrificed.The lungs were removed for determination of wet/dry lung weight ratio (W/D ratio),and for examination of the pathological changes.The kidneys were removed for determination of the expression of claudin-5 and ZO-1 (by immunohischemistry assay and Western blot) and for examination of the pathological changes.The damage to the renal tubules was scored.Results Compared with group C,W/D ratio,plasma Cr and BUN concentrations,and renal tubular damage score were significantly increased,and the expression of claudin-5 and ZO-1 was down-regulated in group ALI (P<0.01).Compared with group ALI,W/D ratio,plasma Cr and BUN concentrations,and renal tubular damage score were significantly decreased,and the expression of claudin-5 and ZO-1 was up-regulated in group H (P<0.01).The pathological changes of lungs and kidneys were significantly attenuated in group H as compared with group ALI.Conclusion Colquhounia root tablet can attenuate kidney injury through up-regulating the expression of tight junction proteins claudin-5 and ZO-1 in the renal tissues of rats with ALI.
2.Effect of heme oxygenase-1 on CDK5-ATM-P53 signal transduction pathway in rat hippocampal neurons subjected to oxygen-glucose deprivation injury
Jianlin SHAO ; Xiaohong WAN ; Weijun ZENG ; Ruhua LONG ; Yan WANG ; Guoliang ZHAO ; Xiaohong GONG
Chinese Journal of Anesthesiology 2012;32(6):732-735
ObjectiveTo investigate the effect of heme oxygenase-1 (HO-1) on cyclin-dependent kinase 5 (CDK5)-ataxia telangiectasia mutated (ATM)-P53 signal transduction pathway in rat hippocampal neurons subjected to oxygen-glucose deprivation (OGD) injury.MethodsHippocampal neurons of newborn Wistar rats ( < 48 h) were cultured for 7 days in vitro.The primary cultured neurons were randomly divided into 4 groups with 10 wells in each group:control group (group C),OGD (group D),OGD + hemin (HO-1 inducer) group (group D + H ) and OGD + hemin + zinc protoporphyrin ( HO-1 inhibitor) group ( group D + H + T).For OGD experiments,cultures were washed three times in a glucose-free balanced salt solution (BSS).They were then placed in deoxygenated glucose-free medium and sealed under 95% N2-5% CO2 in an anaerobic chamber equilibrated to 37°C and 100% humidity for 45 min.OGD was terminated by replacement of stored medium and by returning the cultures to a standard incubator maintained at 37 ℃ in 95% air-5% CO2.The OGD model was established after the neurons were preconditioned with hemin 10 μmol/L for 24 h in group D + H.The OGD model was established after the neurons were preconditioned with hemin 10 μmol/L and zinc protoporphyrin 10 μmol/L for 24 h in group D + H + T.After 24 h of culture,the neuronal viability,apoptosis rate,and expression of HO-1 mRNA and protein,and CDK5,ATM and P53 protein were detected.ResultsCompared with group C,the expression of HO-1 mRNA,and HO-1,CDK5,ATM and P53 protein was up-regulated,the neuronal viability was significantly decreased,and the apoptosis rate was significantly increased in group D (P < 0.01 ).Compared with group D,the expression of HO-1 mRNA and protein was up-regulated,the expression of CDK5,ATM and P53 protein was down-regulated,the neuronal viability was significantly increased,and the apoptosis rate was significanlly decreased in group D + H ( P < 0.01 ).Compared with group D + H,the expression of HO-1 mRNA and protein was down-regulated,the expression of CDK5,ATM and P53 protein was up-regulated,the neuronal viability was significantly decreased,and the apoptosis rate was significantly increased in group D + H + T ( P < 0.01 ).ConclusionHO-1 can inhibit neuronal apoptosis through blocking CDK5-ATM-P53 signal transduction pathway in rat hippocampal neurons subjected to OGD injury.
3.Comparing the effects of transcuataneous electrical nerve stimulation and fluoxetine on central pain in patients with spinal cord injury
Xiaohong WANG ; Bin SHAO ; Qin WANG ; Jingwen XU
Chinese Journal of Physical Medicine and Rehabilitation 2011;33(6):452-454
Objective To compare the effects of fluoxetine and transcuataneous electrical nerve stimulation (TENS) on central pain after spinal cord injury (SCI) using a sham-controlled crossover method.Methods Ele-ven patients with central pain after SCI were randomly divided into two groups which were then subject to 2 phases of treatment.Patients in group 1 were treated by oral intake of fluoexetine for 4 weeks followed by TENS treatment for 4 weeks.Those in group 2 were treated in the reverse sequence.A fifteen day washout period was arranged between the two phases of treatment.The short-form McGill pain questionnaire (SF-MPQ) and the Beck depression inventory (BDI) were used to assess all patients pre-and post-treatment.Results SF-MPQ scores were reduced significantly after either fluoexetine or TENS treatment.After each phase of treatment there was no significant difference between the two groups.Significant improvement in terms of BDI scores was found with fluoxetin treatment in both phases of the trial,but not with TENS treatment.Conclusions Both fluoxetine and TENS can alleviate central pain after SCI,and fluoxetine can relief patients' depression at the same time.
4.Role of HO-1 in inhibition of oxygen-glucose deprivation-induced apoptosis in rat hippocampal neurons by sevoflurane preconditioning
Jianlin SHAO ; Xiaohong WAN ; Yan WANG ; Rongbi HANG ; Xinhua HENG
Chinese Journal of Anesthesiology 2010;30(4):484-487
Objective To investigate the role of HO-1 in inhibition of oxygen-glucose deprivation (OGD)-induced apoptosis in rat hippocampal neurons by sevoflurane preconditioning.Methods Hippoeanlpal neurons of newborn Wistar rats (<48 h) were cultured in vitro.Tne neurons were randomly divided into 6 groups with 108 wells in each group:control group(group C),2% sevoflurane preconditioning group (group S1),OGD group,S1 +OGD group,4% sevoflurane preconditioning+OGD group (group S2+OGD),and 4% sevoflurane preconditioning+ZnPPⅨ+OGD group(group Z).Group C received no treatment.The neurons were cultured for 24 h after 2% sevoflurane preconditioning in group S1.For OGD experiments,the neurons were placed in deoxygenated glucose-free medium and sealed under 95% N2-5% CO2 in an anaerobic chamber equilibrated to 37℃ and 100%humidity for 45 min.then OGD was terminated by replacement of the stored medium and returning the cultures to a standard incubator maintained at 37℃ in 5% C02 and the neurons were cultured for 24 h as described by Ray et al. The OGD model was established after 2% and 4% sevoflurane preconditioning in group S1 + OGD and S2 + OGD respectively. In group Z, when the neurons were preconditioned with 4% sevoflurane, ZnPPⅨ was added to the culture medium at the same time, and the other procedures were the same as those in group S2 + OGD. The neuron viability, apoptesis rate, and expression of HO-I protein and mRNA were detected at 24 h of culture. Results Compared with group C, neuron viability was significantly decreased,apoptosis rate was significantly increased, and expression of HO-1 protein and mRNA was up-regulated in group OGD, S1 + OGD, S2 + OGD and Z, expression of HO-1 protein and mRNA was up-regulated in group S1 ( P < 0.01 ), but no significant change was found in neuron viability and apoptosis rate in group S1 ( P > 0.05). Compared with group OGD, neuron viability was significantly increased, apoptosis rate was significantly decreased, and expression of HO-1 protein and mRNA was up-regulated in group S1 + OGD and S2 + OGD ( P < 0.01), but no significant change was found in the indexes mentioned above in group Z ( P > 0.05 ). Neuron viability was significantly higher, apoptosis rate lower and expression of HO-1 protein and mRNA higher in group S2 + OGD than in group S1 + OGD ( P < 0.01). Neuron viability was significantly lower, apoptosis rate higher and expression of HO-1 protein and mRNA lower in group Z than in group S2+OGD(P<0.01).Conclusion HO-1 is involved in the inhibition of OGD-indueed apoptosis in rat hippocampal neurons by sevoflurane preconditioning.
5.Determination of human papillomavirus types in giant condyloma acuminatum
Xiaohong SHAO ; Yunsheng XU ; Zhiming LI ; Bingxu LI
Chinese Journal of Dermatology 2012;45(5):362-363
ObjectiveTo determine the genotypes of human papillomavirus(HPV) in patients with giant condyloma acuminatum.MethodsSixty-seven outpatients with giant condyloma acuminatum collected from January 2007 to January 2010 were included in this study.Lesional specimens were obtained from these patients.The genotypes of HPV were determined by flow-through hybridization and gene chip assay.ResultsOf the 67 cases of giant condyloma acuminatum,63 (94.02%) were positive for HPV DNA.Among the HPV DNA-positive specimens,84 (60.87%) harbored low risk types of HPV,54 (39.13%) high risk types of HPV.Type 6 and 11were the predominant low risk HPV types,while type 16 and 18 were the major high risk HPV types.
6.Effects of magnetic stimulation on apoptosis of nerve cells and the production of iNOS after spinal cord injury
Bin SHAO ; Xiaohong WANG ; Ning ZHOU ; Qiang ZHANG
Chinese Journal of Physical Medicine and Rehabilitation 2011;33(1):10-13
Objective To investigate effects of magnetic stimulation on apoptosis of nerve cells and the production of inducible nitric oxidate synthase (iNOS) after spinal cord injury (SCI). Methods Thirty-two SpragueDawley male rats were randomly divided into a magnetic stimulation group (n = 16) and a control group (n = 16).SCI models were established by spinal cord transection in both groups. Rats were sacrificed at the 6th, 12th, 24th and 72nd hour post-injury, but the rats in the stimulation group received magnetic stimulation before being sacrificed.Apoptosis index (AI) and iNOS-positive cells rate were recorded at each time point. Results Apoptotic cells could be observed by the 6th hour post-injury, and were elevated from the 24th to the 72th hour. iNOS-positive cells were few at the first two time points, but had increased significantly at the 24th and 72nd hour post-injury. Compared with the control group, the apoptosis index of the stimulation group decreased a little at the 6th and 12th hour, but not significantly. The difference was quite significant at the 24th and 72nd hour, however, and the AI in the stimulation group decreased much more than that in the control group. There was little difference in the rate of iNOS-positive cells between the control and stimulation groups at any time point. Conclusions Magnetic stimulation could inhibit neural apoptosis and protect neurons from secondary SCI, but it has little effect on iNOS production.
7.Oleanolic acid-induced apoptosis and its relation with intracellular calcium in human lung adenocarcinoma A549 cells
Xiaohong WEI ; Aiqun MA ; Jie SHAO ; Junhui WANG ; Asmitanand THAKUR
Journal of Pharmaceutical Analysis 2010;22(2):116-119
Objective To investigate the effect of oleanolic acid (OA) on apoptosis, correlation between apoptosis and intracellular calcium, and its mechanism in human lung adenocarcinoma cell line A549. Methods Human lung adenocarcinoma A549 cells were incubated in vitro and assigned with OA concentrations of 0, 10, 20 and 40μg/mL. The apoptosis status of A549 cell line was detected with Annexin V-FITC/PI by flow cytometry (FCM); fluorescence intensity (FI) of A549 cells was assessed and the level of intracellular calcium was calculated at 24 hour of OA intervention. The relation between apoptosis and calcium FI was illustrated by curve fitting. Results FCM showed that 10, 20 and 40μg/mL of OA could induce A549 cell apoptosis, which followed a concentration-effect pattern; 24-hour intervention with 20μg/mL and 40μg/mL OA showed increased A549 cell apoptosis, and was significantly different from that with 0μg/mL OA (P<0.01). The FI of intracellular calcium concentration in 10, 20 and 40μg/mL OA groups was significantly higher than that in 0μg/mL group after 24 hours' intervention, and the FI showed a trend of increase with increased OA concentration (P<0.01). Curve fitting showed a significant correlation between apoptosis rate and intracellular calcium concentration in A549 cells (r=0.981, P<0.01). Regression equation was Y=0.508X-1.627. Conclusion OA plays a role in inducing apoptosis of human lung adenocarcinoma cells in a concentration-dependent manner. The OA-induced apoptosis is responsible for intracellular calcium overload of the tumor.
8.Multivariate analysis for the factors affecting recurrence of aggressive fibromatosis
Xiaohong NING ; Lin ZHAO ; Yajuan SHAO ; Yuzhou WANG
Basic & Clinical Medicine 2006;0(06):-
Objective To study the factors influencing the recurrence of aggressive fibromatosis(AF) patients.Methods Analyze clinical features and find out factors affecting AF recurrence with univariate and multivariate analysis. Results One hundred and three AF patients were analyzed. Diameter of all the masses was (7.84?5.62)cm. 98.2% of all patients received surgery and radical resection rate is 79.4%. First recurrent time after surgery of male and female is (1563?377) and (2117?3704) days for male and female cases respectively,it's (2723?461),(657?262),(2090?499),(812?220) and (721?234) days for that of abdomen wall、head and neck(H & N),deep mass,joints and chest wall. Recurrent time is (2232?271) and (1347?267) days for those with or without surgery history patients. Univariate analysis showed that gender,tumor site and surgery history of tumor site are prognostic factors of tumor recurrence. Multivariate analysis suggest that only surgery history of tumor site is an independant prognostic factors of AF recurrence after surgery. Conclusion Gender,tumor location and surgery history of tumor site can be valuable to predict postsurgery recurrence of AF.
9.Detecting of gene oipA of normal and clinical Helicobacter pylor strains and comparing of their nucleotide sequences
Shihe SHAO ; Hua WANG ; Muqing LIU ; Xiaohong HAN ; Xiujie DUAN
Chinese Journal of Immunology 1985;0(02):-
Objective:To detect the oipA gene of Helicobacter pylori(Hp)strains NCTC11637 and Hp1,Hp2 isolated from clinical biopsies,analyze their nucleotide sequences and make a homologous comparison of nucleotide with Hp 26695.Methods:The oipA gene was detected with PCR in Helicobacter pylori(Hp)strains NCTC11637 and Hp1,Hp2 isolated from clinical gastric biopsies after routine culture.Then PCR products were sent out for nucleotide sequence analysis and compared with Hp 26695.Results:The sequence of the aim gene was obtained in NCTC11637 and Hp1,Hp2 and was made a homologous comparison of nucleotide with 26695.The number of mutation of NCTC11637,Hp1 and Hp2 and was 48,48,50 respectively.The identity was 94%,94% and 94% respectively,while the strain Hp1 was most identical to 11637 as much as 100%.The homology of Hp2 and 11637 was 97%.Conclusion:Hp1,Hp2 and NCTC11637 expresse gene oipA,but the sequences of gene oipA of different strains are distinct.
10.Effect of transducted heme oxygenase-1 protein on rat brain ischemia-reperfusion injury
Xiaohong WAN ; Yan WANG ; Guoliang ZHAO ; Huamei CHEN ; Jianlin SHAO
Chinese Pharmacological Bulletin 2014;(5):628-631,632
Aim To investigate the effect of transduct-ed-hemeoxygenase-1 ( HO-1 ) protein on brain ischemi-a-reperfusion ( I/R ) rat hippocampal neurons injury. Methods 11 R ( arginine residues )-fused HO-1 pro-tein was established and 50 male Mongolian gerbils were randomly divided into 5 groups ( n=10 ):I/R ( control group ) , I/R + 5 mg · kg-1 saline group ( group S ) , I/R + 5 mg · kg-1 11 R group ( group R), I/R + 5mg·kg-1 11R-HO-1 group (group H1) and I/R + 25 mg · kg-1 11 R-HO-1 group ( group H2). For I/R experiments, ischemia was induced for 5 min by occluding the common carotid arteries bilater-ally with aneurysm clips under Ketamine anesthesia. The experiment was conducted after the neurons were intraperitoneally injected with 5mg·kg-1 saline,11R , 11R-HO-1,or 25mg · kg-1 11R-HO-1 for 3 h. The rats were killed after 24h of reperfusion. Hippocampus was removed immediately for determination of cAMP level, neuronal apoptotic rate, and expression of HO-1 and Caspase-3 protein, mitochondria was observed un-der electron microscope. Results Among group C, group S and group R,there were no differences in the expressions of HO-1, Caspase-3 protein, cAMP level , neuronal apoptotic rate and mitochondria damage ( P>0. 05). Compared with group C, group S and group R, the expression of HO-1 protein was up-regulated, the expression of Caspase-3 protein was down-regula-ted, cAMP level increased, the apoptotic rate was sig-nificantly decreased and mitochondria damage de-creased in group H1 ( P < 0. 01 ) . Compared with group H1 , the expression of HO-1 protein was up-regu-lated, the expression of Caspase-3 protein was down-regulated, cAMP level increased, the apoptotic rate was significantly decreased and mitochondria damage decreased in group H2 ( P <0. 01 ) . Conclusion Transducted-HO-1 protein can attenuate brain ischemi-a-reperfusion rat hippocampal neurons injury.