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【Objective】 To explore the expression of USP9X in platelets and its effect on platelet function. 【Methods】 The expression of USP9X in human and mouse was evaluated by PCR and Western blot. Platelets from young and old mice were separated and prepared, and the expression of USP9X was detected. USP9X inhibitos were used to assess the regulation of USP9X in platelet function, including aggregation, ATP release and spreading. Platelet lysates were collected in different time points to evaluate the change of phosphorylation of Akt in USP9X inhibitors treated platelets. 【Results】 Both human and mouse platelets expressed USP9X. Compared to the young mice, the old mice showed significantly enhanced expression of USP9X(P<0.05). To assess the effect of USP9X on platelet function, USP9X inhibitor was used to pre-incubate platelets for 30 min and platelet function were examined later. Results showed that USP9X inhibitor significantly decreased platelet activation including aggregation, ATP release and spreading(P<0.05). Compared to the control group, the inhibitor treated group showed a significant decrease in the spreading area after 45 minutes. The Western blot results showed a significant decrease in Akt phosphorylation levels of platelets in the USP9X inhibitor treated group. 【Conclusion】 Both human and mouse platelet express USP9X, and inhibition of USP9X decreased platelet function including aggregation, ATP release and spreading. USP9X can also influence the phosphorylation of Akt. The inhibitor of USP9X may become a potential therapeutic target for thrombosis intervention.
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ObjectiveTo explore the mechanism of Xianglian Huazhuo prescription in the treatment of chronic atrophic gastritis (CAG) based on network pharmacology and animal experiments,so as to provide scientific basis for clinical application. MethodThe possible targets and pathways of Xianglian Huazhuo prescription in the treatment of CAG were obtained based on the prediction of network pharmacology. The CAG rat model was induced by sodium salicylate,N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) and hunger and satiety disorder. Then the CAG rats were treated with Xianglian Huazhuo prescription and morodan for 60 days. After administration,the rats were sacrificed,and the content of interleukin-6 (IL-6),tumor necrosis factor-α (TNF-α),interleukin-1β (IL-1β) and vascular endothelial growth factor (VEGF) in serum was determined by enzyme linked immunosorbent assay(ELISA). In addition, the protein expression of Bad and Bcl-2 in gastric mucosa was detected by immunohistochemistry (IHC). ResultA total of 241 active components of Xianglian Huazhuo prescription and 53 core targets were obtained. Xianglian Huazhuo prescription affected multiple biological processes,such as cell proliferation and apoptosis,inflammatory reaction,regulation of DNA metabolism,and cell response to redox,as well as phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt),TNF,mitogen-activated protein kinase (MAPK),cancer and cancer-related signaling pathways. The animal model verification showed that Xianglian Huazhuo prescription lowered the levels of IL-6,TNF-α,IL-1β and VEGF in serum of CAG rats,and reduced the protein expression of Bad and Bcl-2 in gastric tissue. ConclusionXianglian Huazhuo prescription could regulate PI3K/Akt signal pathway and improve gastric mucosal injury in CAG by participating in biological processes such as cell proliferation,apoptosis and inflammation.
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ObjectiveTo explore the mechanism of Xianglian Huazhuo prescription in the treatment of chronic atrophic gastritis (CAG) based on network pharmacology and animal experiments,so as to provide scientific basis for clinical application. MethodThe possible targets and pathways of Xianglian Huazhuo prescription in the treatment of CAG were obtained based on the prediction of network pharmacology. The CAG rat model was induced by sodium salicylate,N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) and hunger and satiety disorder. Then the CAG rats were treated with Xianglian Huazhuo prescription and morodan for 60 days. After administration,the rats were sacrificed,and the content of interleukin-6 (IL-6),tumor necrosis factor-α (TNF-α),interleukin-1β (IL-1β) and vascular endothelial growth factor (VEGF) in serum was determined by enzyme linked immunosorbent assay(ELISA). In addition, the protein expression of Bad and Bcl-2 in gastric mucosa was detected by immunohistochemistry (IHC). ResultA total of 241 active components of Xianglian Huazhuo prescription and 53 core targets were obtained. Xianglian Huazhuo prescription affected multiple biological processes,such as cell proliferation and apoptosis,inflammatory reaction,regulation of DNA metabolism,and cell response to redox,as well as phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt),TNF,mitogen-activated protein kinase (MAPK),cancer and cancer-related signaling pathways. The animal model verification showed that Xianglian Huazhuo prescription lowered the levels of IL-6,TNF-α,IL-1β and VEGF in serum of CAG rats,and reduced the protein expression of Bad and Bcl-2 in gastric tissue. ConclusionXianglian Huazhuo prescription could regulate PI3K/Akt signal pathway and improve gastric mucosal injury in CAG by participating in biological processes such as cell proliferation,apoptosis and inflammation.
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Objective To explore the effects of diagnostic delay on early disease course of Crohn's disease (CD ), and to analyze the risk factors related with the progress of early disease course. Methods From December 2014 to July 2017,a total of 56 patients in Renmin Hospital of Wuhan University initially diagnosed as CD were selected.The clinical manifestation,previous history,imaging examination, endoscopic examination and pathological results of the patients were collected after initial diagnosis of CD. CD-related surgery,CD-related rehospitalization and use of immunomodulators were followed.According to time interval between initial symptom onset and initial diagnosis,the patients were divided into diagnostic delay group (diagnostic delay time>two years,14 cases)and non-diagnostic delay group (diagnostic delay time≤two years,42 cases).The clinical characteristics at initial diagnosis and early prognosis were compared between the two groups.The risk factors affecting early prognosis of CD patients were analyzed. Kaplan-Meier was used for prognosis analysis.Log-rank test and Cox proportional hazards model were performed to analyze factors affecting early prognosis.Results The results of imaging examination indicated that the proportion of intestinal injury of diagnostic delay group was significantly higher than that of non- diagnostic delay group (χ2= 4.49,P = 0.03 ).During follow-up,the proportion of CD-related rehospitalization of diagnostic delay group was higher than that of non-diagnostic delay group,and the difference was statistically significant (χ2=7.34,P=0.01).During follow-up,the nine patients received surgery.The one-year and three-year cumulative incidence of surgery was 10.7% and 22.7%,respectively;one-year cumulative incidence of surgery in diagnostic delay group and non-diagnostic delay group was 38 .6%and 9 .6%,respectively.The results of Cox univariable and multivariable analyses both indicated that intestinal injury in imaging examination at diagnosis was the risk factor of surgery (both hazard ratio (HR)=2.313,both P<0.01).A total of 21 patients had CD-related rehospitalization during follow-up.And one-year and 31-month cumulative incidence of hospitalization was 27 .8% and 5 6 .3%,respectively;one-year cumulative incidence of CD-related rehospitalization in diagnostic delay group and non-diagnostic delay group was 43 .7% and 2 1 .6%,respectively;and the difference was statistically significant (χ2=5 .88 ,P=0.02).The results of Cox univariable analysis indicated that having systematic manifestation,intestinal inj ury in imaging examination and diagnostic delay were the risk factors of CD-related rehospitalization (HR=2.972,2.313 and 1.403,all P<0.05).The results of multivariable analysis indicated that having systematic manifestation and intestinal injury in imaging examination were independent risk factors of CD-related rehospitalization (HR=3.891 and 2.541, P < 0.01,respectively).Conclusions The proportion of intestinal injury of patients with diagnostic delay is high and the early prognosis is poor. Intestinal injury in imaging examination at initial diagnosis is independently risk factor of CD-related surgery and rehospitalization during follow-up.
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OBJECTIVE@#To study the effect of Wdr1 deletion in germ cells on ovarian function of mice.@*METHODS@#Oocyte-specific gene knockout mouse model was constructed by crossing Wdr1female mice with Cre recombinase transgenic male mice which was driven by a germ cell-specific promoter. Wdr1; Ddx4-Cre mice and control mice were sacrificed at 14 days, 28 days and 4 months after birth, whose ovaries were subjected to photography, paraffin sectioning and Hematoxylin-Eosin (HE) staining. The ovarian volume and follicular numbers were recorded at various time points.@*RESULTS@#The ovarian volume of Wdr1 ; Ddx4-Cre mice was slightly lower than that of the controls at 14 days. HE staining showed that primordial follicles, primary follicles and secondary follicles were slightly reduced compared with the control mice at 14 days. The ovarian volume of Wdr1 ; Ddx4-Cre mice was significantly lower than that of the control mice at 28 days and 4 months. HE staining showed that all developmental follicles were significantly reduced compared with the control mice.@*CONCLUSION@#Wdr1 gene deletion in germ cells can influence early ovarian function of mice and lead to premature ovarian failure.
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Animales , Femenino , Masculino , Ratones , Células Germinativas , Ratones Noqueados , Ratones Transgénicos , Proteínas de Microfilamentos , Genética , Oocitos , Folículo OváricoRESUMEN
Objective To analyze the infection status and possible risk factors of Helicobacter pylori among people taken physical examination in Miyun area of Beijing,so as to establish effective intervention measures scientifically.Methods From January 2012 to December 2016,eleven thousand physical examinees in Beijing MIyun Hospital were enrolled in this study,13C-urea breath test was used to detect Helicobacter pylori infection and a face to face questionnaire survey was applied.The data colleted was analyzed based on sex,age, living habits.Results The overall infection rate of Hp was 46.72%(5139/11000),the rate of male infection was higher than that of female(49.74%(2826/5682)vs.43.49%(2313/5318)),and the difference was statistically significant(χ2=43.000,P<0.000).The physical examinees were divided into 4 groups by 20 years for age.The infection rates of Hp in the<20 years old group,20—39 years old group,40-59 years old group,>60 years old group were 36.17%,47.28%,48.04%,43.59%,respectively.The infection rate before the age of 60 was on the rise,and the infection rate was the highest in the 40-59 year group,the lowest in the<20 years old group.There was significant difference among the 4 groups(χ2=23.694,P=0.000).The Hp infection rate in people who smoked,had a large number of family members and preferred a hot diet,a dinner party and no hand washing habits was significantly higher than those without related habits,the differences were statistically significant(P<0.05).Conclusion The infection rate of Hp infection in the MIyun area is lower than that of the national average.The infection distribution is closely related with sex,age and living habits.Health education should be strengthened and the occurrence of food-borne disease should be prevented to reduce the Hp infection rate.
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Objective To analyze the infection status and possible risk factors of Helicobacter pylori among people taken physical examination in Miyun area of Beijing,so as to establish effective intervention measures scientifically.Methods From January 2012 to December 2016,eleven thousand physical examinees in Beijing MIyun Hospital were enrolled in this study,13C-urea breath test was used to detect Helicobacter pylori infection and a face to face questionnaire survey was applied.The data colleted was analyzed based on sex,age, living habits.Results The overall infection rate of Hp was 46.72%(5139/11000),the rate of male infection was higher than that of female(49.74%(2826/5682)vs.43.49%(2313/5318)),and the difference was statistically significant(χ2=43.000,P<0.000).The physical examinees were divided into 4 groups by 20 years for age.The infection rates of Hp in the<20 years old group,20—39 years old group,40-59 years old group,>60 years old group were 36.17%,47.28%,48.04%,43.59%,respectively.The infection rate before the age of 60 was on the rise,and the infection rate was the highest in the 40-59 year group,the lowest in the<20 years old group.There was significant difference among the 4 groups(χ2=23.694,P=0.000).The Hp infection rate in people who smoked,had a large number of family members and preferred a hot diet,a dinner party and no hand washing habits was significantly higher than those without related habits,the differences were statistically significant(P<0.05).Conclusion The infection rate of Hp infection in the MIyun area is lower than that of the national average.The infection distribution is closely related with sex,age and living habits.Health education should be strengthened and the occurrence of food-borne disease should be prevented to reduce the Hp infection rate.
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Objective To package the recombinant lentivirus containing HIV-1 Vpr gene and detect the effect of Vpr protein expression on the latent infection and lyric replication of KSHV.Methods The fragment of Vpr gene from expression plasmid pCI-neo-Vpr was cloned into the lentivirus vector pHAGE-CMV-MCS-IzsGreen,then the recombinant plasmid pHAGE-Vpr,package vector psPAX2 and envelope vector pMD2.G were cotransfected into 293T cells.GFP expression was observed by fluorescent microscopy.Culture media of 293T cells were harvested and filtered through 0.45 μm filter.After 293T cells were infected by a series of diluted lentivirus,the virus titer was checked by observing GFP expression.Vpr mRNA transcripts in 293T cells were detected by RT-PCR.Then BCBL-1 cells were infected by the recombinant lentivirus with 1 MOI,GFP expression was observed by fluorescent microscopy,and the mRNA transcripts and protein expression of Vpr in BCBL-1 cells were detected by RT-PCR and Western blot.Meanwhile,the mRNA transcripts and protein expression of KSHV lytic cycle gene Rta were detected by RT-PCR and Western blot,respectively.Results The recombinant lentivirus carrying HIV-1 Vpr was packaged successfully with the virus titer of 4 × 107 TU/ml.After infected with lentivirus,BCBL-1 cells could express GFP,and the exact band of Vpr was detectable by RT-PCR and Western blot.Moreover,the expression of KSHV Rta mRNA and protein were downregulated by Vpr protein.Conclusion Overexpression of HIV-1 Vpr mediated by the recombinant lentivirus could inhibit KSHV lytic replication and enhance KSHV latent infection.
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Objective To investigate the effect of apolipoprotein E gene knock-out(ApoE KO) and high-fat diet on morphology and the expression of mortalin in hippocampal CA_3 neurons of mice, and to explore the impact of these factors on memory and Alzheimer's disease.Methods Ten wild-type and 10 ApoE KO mice were fed with common chow as the control group and the KO group respectively while 10 ApoE KO mice were fed with high fat diet.Twelve weeks later, the weight and the lid of these mice were measured.The brain tissues were observed using HE staining, nissl staining, protargol staining,immunohistochemistry staining and image analysis by computer.Results In the ApoE KO group, weight,total cholesterol, triglyceride, low-density lipoprotein cholesterol were higher than those in the control group,and these changes were more significant in ApoE KO high-fat diet group.The nissl was higher in the ApoE KO group (0.301±0.031) and in ApoE KO high-fat diet group (0.261±0.020) than those in the control group (0.341±0.035, F=18.068, P<0.05).The mortalin in the ApoE KO group (0.322±0.060) and in ApoE KO high-fat diet group (0.391±0.041) were higher than the control group (0.256±0.061, F=15.230, P < 0.05).Conclusions ApoE KO and high-fat diet can reduce nissl, and improve the expression of mortalin.This protein may be involved in the pathogenesis of Alzheimer's disease.
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Objective To study the effect of hypothemah on the early inflammatory reaction in acute lung injury induced by intestinal ischemia-repeffusion(IlR)in rabbits.Method Seventy-two healthy rabbits provided by Peking Union Medical Colege Hospital Anhnal center were randomly divided into four groups(n=18 pergroup):(1)normothermia control group (rectal temperature 37-38 C;sham group);(2)normothermia IlR group(rectal temperature 37-38 C);(3)mild hypothermia HR group(rectal temperature 32-35℃);and (4)moderate hypothermia IIR group(rectal temperature 28-31.9C).Acute lung injury was induced by claIllp.ithe superiornteric artery(SMA)for 1 hour and declamping the SMA for 6 hours.Hypothermia WaS induced by surface cooling.Before and 2.4 and 6 hours after IIR,the Olasmlevels o,IL-,IL-6 and IL10 were measured.All rabbits were killed 6 hours after IIR and water content in lung tissue Wttk'assessed.Iaght mieropic examination was performed tbr morphological assessment of the hmg.The data were analyzed by AN()VA.Statistical significance wag dned as a P of<0.05.Results In the IIR groups,the plasma levels ofTHE-a.IL-l,IL-6 and IL-10 and lung water were increased.There Was evidence of acute lung injury from morphologi-cal assessment of the lung.The acute lung injury induced by IIR was improved by hypethennia.Mild hypothermia Was similar to moderate hypothermia for the treatment of acute lung injury induced by IIR.ConclusiotMild hy-pothermia and moderate hypothermia Can significantly improve acute lung injury induced by IIR in rabbits.Mild hypothea had similar efficacy to moderate hypothermia for the treatment of acute lung injury induced by IIR.
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Objective To determine the content of indirubin in Kangbingan Oral Liquid by UV spectrophotometer methods.Methods The content of indirubin was determined by Heltos Gamma ultravioletvisible spectrophotometer with chloroformic solution of 2~8 ?g/mL confected by standard sample of indigotin and indirubin.The sample of negative blank solution was confected by the prescription without Radix Isatidis.The standard sample and the blank sample were determined separately with the wavelength at 500~700 nm.Results Indirubin had the maximum absorption at wavelength of 540.0 nm and indigotin had the maximum absorption at wavelength of 634.5 nm.The linear correlation was available at the range of 0.20~1.40 ?g/mL,and the correct coefficient was 0.999 4.The average recovery rate was 99.05% and RSD was 0.87%(n=6).Conclusion This method is simple and accurate,and can be used to control the quality of Kangbingan Oral Liquid.
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Objective To study the location and significance of leptin and leptin receptor in rat submanibular gland. Methods Submandibular gland from SD rats were observed by hematoxylin-eosin, leptin and leptin receptor immunohistochemical staining. Result The granular convoluted tube and striated duct were leptin and leptin receptor positive. The density of positive cells in granular convoluted tube were much high than those of in striated duct. The acini were negative.Conclusion leptin and leptin receptor were widly expressed in granular convoluted tube and striated duct. These results might indicate that submandibular gland participated in adjusting the balance of energy.;
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AIM: To study the possibility and conditions of transplacental infection of coxsackievirus B3(CVB 3) from pregnant mice to their fetuses and newborns. METHODS: Coxsackievirus B 3 strain causing balb/c mice myocardial injury(CVB 3m )was inoculated with 10 5 TCID 50 in dose into the mother mice at 6-7 days (early gestation),9-10 days (middle gestation) and 17-18 days (late gestation) of gestation, in contrast with non pregnant mice. Some placentas and fetuses were removed by caesarean section before mothers partusing; some mothers and their babies were sacrificed after parturition, and virus isolation, serological and pathological tests were performed. RESULTS: Viramiae was observed in mother mice of late gestation inoculated with CVB 3m at a fit amount on the second day after inoculation, while no newtralizing antibody to CVB 3m was detected in blood. The virus was isolated from cardiac muscles of inoculated mother mice in different gestation and the controls. The virus was also isolated from some placentas and fetuses, and both sera and cardiac muscles of infants in the late gestation (virus titer were all 10 -2 -10 -3 ). On d 7 of inoculating virus, pregnant and non pregnant mice titers of neutralizing antibody to CVB 3m in sera were all between 1160 and 1320. Under the electromicroscopy, some cardiac muscle cells of mother or infant mice appeared with morphological changes and little hollow bubbles occured in cytoplasm. The fibers broke off, and the bright and dark belts became indistinct. CONCLUSION: The amimal model, intraplacental passage of CVB 3 from pregnant mother in late gestation to fetus in mice, is a benefitial tool to study enterovirus diseases in human perinatal period.
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0.05).The eggs per gram feces and liver, eggs in uterus per female, and egg granulomas on the liver surface were(56.68?24.78),(5 826?437),(49.94?12.53) and(10.04?1.13)/0.25 cm2, respectively in immunized group, while in control group these were(89.93?32.18),(10 016?3 541),(76.54? 19.77) and(19.22?2.45)/0.25 cm2 respectively, all with significant difference(P
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Aim To compare cardioprotection effects between carvedilol and metoprolol in canine late reperfusion of acute myocardial infarction.Methods Eighteen anesthetized dogs were randomly divided into three groups: late reperfusion group(LR,n=6),late reperfusion after metoprolol treatment group(LR+M,n=6),and late reperfusion after carvedilol treatment group(LR+C,n=6),respectively orally giving physiological saline,metoprolol(1 mg?kg~(-1)?d~(-1)),and carvedilol(1 mg?kg~(-1)?d~(-1)) for seven days,and then late reperfusion of acute myocardial infarction model was made by ligating the coronary for 6 h,followed by reperfusion for 6 h.SOD,GR activity and MDA content of infarction brim myocardium were detected by colorimetry,Fas/FasL were detected by immunohistochemistry,apoptosis index(AI) were detected by TUNEL.Results Compared with LR,Myocardial MDA content in LR+C was decreased,and SOD and GR activities were significantly higher,but LR+M did not change.The expression of Fas/FasL and apoptosis index were significantly lowered in LR+M and LR+C,especially in LR+C.Conclusion Carvedilol and metoprolol have cardioprotection on late reperfusion of acute myocardial infarction,and carvedilol is superior to metoprolol and the pharmacological effects may due to its antioxidant effect.
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Objective To observe the distribution of the neuropeptides in mouse sublingual gland. Method Using immunohistochemical ABC method. Results The striated duct cells in the sublingual gland showed somatostatin and calcitonin gene related peptide immunoreactivity positive which were in the cytoplasma, but the nucleus negative.Conclusion The striated duct cells in mouse sublingual gland also contained neuropetides; which may play an important role in regulating acinar secretion and blood supply.