AIM:To investigate the effect of caveolin-1 and phosphorylation of ERK1/2 on 17?-estradiol (E2) induced inhibition of vascular smooth muscle cells (VSMCs). METHODS:The proliferation in cultured VSMCs was determined by using [3H]-thymidine incorporation. The expressions of caveolin-1,MKP-1 and ERK1/2 phosphorylation were measured by Western blotting. The expression of caveolin-1 mRNA was measured by RT-PCR. RESULTS:Exposed to fetal calf serum (FCS) for 24 h,the increase in proliferation of VSMCs was detected by [3H]-thymidine incorporation. Pretreatment with various concentrations of E2 for 24 h inhibited VSMC proliferation induced by FCS. The results of Western blotting and RT-PCR showed that pretreated with 17?-estradiol for 24 h reserved the decrease in caveolin-1 induced by FCS. Western blotting results further proved that the expression of MKP-1 was significantly increased and the expression of ERK1/2 phosphorylation was decreased after incubated with 17?-estradiol. CONCLUSION:17?-estradiol increases caveolin-1 and MKP-1 expressions,and decreases ERK1/2 phosphorylation,leading to the inhibition of VSMC proliferation.

AIM: To investigate the effect of caveolin - 1 and phosphorylation of ERK1/2 on 17β - estradiol ( E_2 ) induced inhibition of vascular smooth muscle cells ( VSMCs ). METHODS: The proliferation in cultured VSMCs was determined by using [~3H ] - thymidine incorporation. The expressions of caveolin - 1, MKP -1 and ERK1/2 phosphorylation were measured by Western blotting. The expression of caveolin - 1 mRNA was measured by RT - PCR. RESULTS: Exposed to fetal calf serum ( FCS) for 24 h, the increase in proliferation of VSMCs was detected by [~3H] -thymidine incorporation. Pretreatment with various concentrations of E_2 for 24 h inhibited VSMC proliferation induced by FCS. The results of Western blotting and RT - PCR showed that pretreated with 17β - estradiol for 24 h reserved the decrease in caveolin - 1 induced by FCS. Western blotting results further proved that the expression of MKP - 1 was significantly increased and the expression of ERK1/2 phosphorylation was decreased after incubated with 17β - estradiol. CONCLUSION: 17β -estradiol increases caveolin - 1 and MKP - 1 expressions, and decreases ERK1/2 phosphorylation, leading to the inhibition of VSMC proliferation.

The prevalence of inflammatory bowel disease (IBD) is rising in recent years. The peak incidence of IBD is mostly occurred on middle and young aged people, which might influence the development of their social and psychological health. The mechanism of IBD related psychological disorder is unknown, however, several studies indicate that the intervention of gut microbiota might improve the psychological symptom and life quality of IBD patients, and reduce the recurrence rate. This article reviewed the role of gut microbiota in IBD related psychological disorder.

Objective:To investigate the expression level and clinical value of serum miR-148a-3p and miR-551b-5p in patients with acute pancreatitis (AP).Methods:The clinical data of 152 patients with AP admitted to Danzhou people's Hospital from January 2017 to September 2020 were selected. According to the severity of their illness, they were divided into MAP group ( n=70), MSAP group ( n=40), and SAP group ( n=42). The SAP group was divided into survival group ( n=25) and death group ( n=17) according to the prognosis of the patients. Another 50 healthy people were selected as the control group. Fastening venous blood was collected on the day of onset, and the expression levels of serum miR-148a-3p and miR-551b-5p in each group were detected by real-time quantitative PCR. The receiver operating characteristic curve (ROC) was drawn and the area under the curve (AUC) was calculated. The expression levels of serum miR-148a-3p and miR-551b-5p were analyzed to evaluate the prognosis of SAP. The correlation between serum miR-148a-3p and miR-551b-5p expression levels in SAP patients was analyzed by Pearson method. Results:The expression levels of serum miR-148a-3p (3.18±1.27 vs 0.96±0.28) and miR-551b-5p (1.94±0.85 vs 0.51±0.12) in AP group were significantly higher than those in control group ( P<0.001). The expression levels of serum miR-148a-3p (4.36±1.70 vs 2.84±1.10, 2.50±0.92) and miR-551b-5p (2.80±1.04 vs 1.68±0.53, 1.42±0.45) in SAP group were significantly higher than those in MSAP group and MAP group ( P<0.001). The expression levels of serum miR-148a-3p (5.30±1.95 vs 3.47±1.40) and miR-551b-5p (3.62±1.37 vs 2.08±0.91) in death group were significantly higher than those in survival group ( P<0.001). ROC curve analysis showed that the AUC of the combination of miR-148a-3p and miR-551b-5 in judging the prognosis of SAP was significantly higher than that of the single index of miR-148a-3p and miR-551b-5[0.943(95% CI0.886-0.997) vs 0.860(95% CI0.797-0.924), 0.822(95% CI0.795-0.880)], with a sensitivity of 98.3% and specificity of 84.6%. Correlation analysis showed that there were positive correlation between the expression level of serum miR-148a-3p and miR-551b-5p in SAP patients ( r=0.835, P<0.001). Conclusions:The expression levels of miR-148a-3p and miR-551b-5p in serum of AP patients were significantly increased, and the combined detection of miR-148a-3p and miR-551b-5p had a good value in judging the prognosis of SAP.

Objective To explore the role of ERK1/2 protein in development of myocardial hypertrophy.Methods Myocardial cells were isolated from ventricles of 1～3-day-old neonate rats and purifed by a culture method.Neonate rat cardiomyocyte hypertrophic responses were assayed by measuring protein content,protein synthesis rate and cell surface area.Expression of protein ERK1/2 were detected by Western blot.Results Cell protein content,~3H-leucine(~3 H-Leu)incorporation and cell surface area increased by treating of cardiomyocytes with T(10~(-10)～10~(-6) mol/L)for 24 h.The maxium effect was observed at the concentration of 10~(-8) mol/L.The increase of cell protein content induced by T was inhibited by pretreating with flutamide(10~(-5) mol/L)for 2 h,while there was no effect on cardiomyocytes pretreating with flutamide alone.The increase of ~3H-Leu incorporation induced by T was blocked by PD98059(50 μmol/L).Expression of ERK1/2 was upregulated significantly by treating with testoster one for 24 h at the level of 10~(-8) mol/L.The increased expression of ERK1/2 induced by T was reversed by pretreating with flutamide(10~(-5) mol/L)for 2 h.Conclusion T with physio-concentration may induce cardiomyocyte hypertrophy and this effect was possibly mediated through the activation of ERK1/2 signalling.During this procession,T upregulated the protein expression of ERK1/2 mediated by androgen receptor.

Objective To investigate the potential roles of dexamethasone (Dex) in podocyte motility,and to explore the mechanism of modulating α-actinin-4,nephrin.Methods Podocytes were divided into three groups:Dex group [1 μmol/L Dex +50 mg/L puromycin aminonucleoside (PAN)],PAN group (50 mg/L) and normal control group.Scrape wound assay and Transwell migration assay were used to detect cell motility.Filtering ratio of podocytes was measured by FITC labeled BSA.Real-time PCR and Western blotting were used to examine the expressions of c-actinin-4 and nephrin.Results From the scrape wound assays,the ability of wound repair in podocytes of PAN group was significantly increased (P＜0.01),and the number of migrating cells in this group also rose (P＜0.01).Compared to PAN group,podocytes in Dex group did not enhance the motility after treatment with the same dose PAN (P＜0.01).Real-time PCR and Western blotting showed that Dex could significantly inhibit the up-regulated expression of α-actinin-4 and nephrin induced by PAN.Conclusions Dex can relieve the enhanced motility induced by PAN.Its mechanism may be associated with the modulation of the expressions of α-actinin-4 and nephrin.

AIM:To investigate the effect of resveratrol on the lipids ( CHOL, TG, LDL-C and HDL-C) , ni-tric oxide ( NO) , peroxynitrite anion ( ONOO-) and the expression of inducible nitric oxide synthase ( iNOS) in the artery of the mice with ovariotomy ( OVX) .METHODS:The lipid levels and NO level in the serum were measured .The chan-ges of atherosclerosis were evaluated with Oil Red O staining .The expression of iNOS was measured by DAB staining and Western blot .The ONOO-production was measured by DAB staining .RESULTS:Compared with sham group , the levels of the lipids and NO production in OVX +high fat (HF) group were increased (P<0.05).Compared with OVX+HF group, the levels of the lipids and NO production in resveratrol group were decreased (P<0.05).Fourteen weeks later, the atherosclerosis model was successfully established .Compared with OVX +HF group, the iNOS expression and the ONOO-production in resveratrol group were decreased ( P<0.05 ) , while those in sham group were increased ( P <0.05).CONCLUSION:Resveratrol prevents and treats atherosclerosis by inhibiting the iNOS expression in C 57BL/6J mice.

Objective To investigate the dynamic expression of nitric oxide synthase(NOS) in the apoptosis of primary cultured rat cortical neruons following hypoxia/reoxygenation(H/R) and the protective role of extract of ginkgo biloba(EGB). Methods The cortical neurons of E16-17 days fetal rat were primarily cultured.The apoptosis model of primary cultured cortical nurons following H/R was established by using W-G staning,electromicroscopy,TUNEL staining.The dynamic expression of NOS different H/R times was investigated with NADPH-diaphorase histochemical method. Results H/R can cause apoptosis of primary cultured rat cortical neurons.In the experiment of H-2R-0,H-4R-0, H-6R-0,H-8R-0 and H-2R 18,H-4R 18,H-6R 18 H-8R 18,the apoptosis cells occurred after 4 hour hypoxia.The increasing of apoptosis cell acted as time-dependence and the peak value was at H-8R 18.The expression of NOS increased both after 2 hour hypoxia and reoxygenation 18 hour after 8 hour hypoxia compared with the normal control group.EGB could inhibit the increasing and decrease the percentage of apoptosis.Conclusion The apoptosis of primary cultured rat cortical neurons could be induced by H/R.The increasing of NO might be one of the mechannisms of apoptosis.EGB could singnificantly inhibit the apoptosis by means of inhibiting the expression of NOS and reducing the production of NO.;

BACKGROUND:In 1990s,overseas researchers use balloon occlusion method for establishing closed-chest animal models of myocardial infarction. But,ventricular fibrillation and thrombosis of intraoperative factors reduce the success rate of establishing the models. Currently,there are a few reports on establishing the large animal models. OBJECTIVE:We used balloon occlusion method for establishing closed-chest swine models of myocardial infarction,and explored ways to improve the success rate of modeling. DESIGN,TIME AND SETTING:The randomized controlled animal study of pathology observation was performed at the Department of Cardiology,First Affiliated Hospital of Kunming Medical College and Research Room of Pathology,Kunming Medical College from July 2008 to May 2009. MATERIALS:Fifteen Diannan small-ear pigs weighing 19-25 kg,aged 8-11 months,were divided into three groups:sham operation group,ischemia-reperfusion group,and ischemic postconditioning group,with 5 pigs in each group.METHODS:After the coronary occlusion and reperfusion period,the prophylactic use of lidocaine (1.0-2.0 mg/kg) infusion to control arrhythmia,and use of heparin to prevent and treat the thrombosis. A balloon catheter was positioned in the distal end of the first diagonal branch of the left anterior descending (LAD) artery under fluoroscopic guidance. In the sham operation group,the balloon was only placed to the LAD,did not block coronary artery. In the ischemia-reperfusion group,inflatable balloon occlusion was done for 60 minutes in the LAD after the balloon removed. In ischemic postconditioning group,after the balloon was inflated and occluded the LAD for 60 minutes,ischemic postconditioning was elicited by eight cycles of 30-second reperfusion,followed by 30-second reocclusion.MAIN OUTCOME MEASURES:Coronary angiography,electrocardiogram (ECG) and cardiac enzymes test was conducted to evaluate models of myocardial infarction. After three days,cardiac 2,3,5-triphenyltetrazolium chloride (TTC) staining and pathological examination was done to verily myocardial infarction.RESULTS:In the sham operation group,all pigs survived. In the ischemia-reperfusion group,4 pig models of myocardial infarction were successfully established,and one died of refractory ventricular fibrillation. In the ischemic postconditioning group,models of myocardial infarction after ischemia were successfully established. Following distal left anterior descending artery occlusion,the ECG leads V13 on the ST-segment elevation,the sick rational Q-wave formed;myocardial enzyme evolution of myocardial infarction in the human body was basically the same process. The site of myocardial infarction,basically the same parts,was located in apical,left ventricular anterior wall,and the former interval. TTC staining was normal myocardium brick red,myocardial infarct area appeared pale;pathological examination revealed a normal structure of myocardial infarct damage,cytoplasm condensed,dyeing deepening,transverse striations disappeared,nuclear enrichment,dissolution,fragmentation,many erythrocytes around the infarct area with abundant granulation tissue and a large infiltration of inflammatory cells.CONCLUSION:The described model presents a less invasion to the animals,and is the closest to the process of clinical practice.Intraoperative use of lidocaine and heparin for controlling arrhythmia and thrombosis of the models is successfully established as an effective manner. Ischemic postconditioning may be one of the factors for improving the modeling success rate.

Objective To summarize the treatment experience of refractory systemic juvenile idiopathic arthritis (JIA) by tocilizumab, and to explore the cost-effective treatment. Methods The clinical data of 6 pediatric patients with refractory systemic JIA treated by tocilizumab from 2014 to June 2016 were retrospectively analyzed in the aspects of course and effectiveness of tocilizumab, steroid reduction, adverse reaction, and growth. Results The median age of the six patients (3 males and 3 females) was 6 years, and the course of disease were from 16 to 63 months. All patients were treated by other immunosuppressive agents or biological agents in addition to steroid and traditional anti-rheumatic drug therapy. The courses of tocilizumab treatment were from 7 to 26 months and the median time was 9.5 months. All 6 patients responded to tocilizumab and achieved the clinical remission at different time. After the induced remission, the interval of the treatment intervention was increased from 2 weeks up to 4 weeks in 3 cases, and no disease activity was observed. Except one case, another 5 cases reduced and stopped the use of hormones at 5.8 months after tocilizumab treatment. After hormones was reduced and discontinued, the growth was improved. All 6 patients had no serious adverse reactions. Conclusions Tocilizumab is safe and effective for patients with refractory JIAs. The steroid can be reduced in short time to improve growth. After remission is induced, the interval of the treatment intervention could be prolonged.