Objective To investigate the possibility of microsatellite alteration (MA) in diagnosis of bladder cancer of Chinese people, and find the better panel which will be used in clinic. Methods A total of 6 and 10microsatellite markers were chosen, PCR-SSLP silver staining assay was performed in 31 and 32 bladder cancers tissue,exfoliate cells in urine and 10, 15 non-bladder cancers exfoliate cells in urine, respectively. Results MA (+) was found in 28 out of 31, 30 out of 32 bladder cancers, and the sensitivity was 90.3%, 93.7% respectively. The MA of urine sediment of 25 non-bladder cancers was negative, and the specificity was 100%. The cytology was carried out among 19 out of 31, 20 out of 32 bladder cancers at the same time, 2 cases ( 10.3 %) and 3 cases ( 15 % ) were found cancer positive, and the sensitivity is significantly lower than that by the analysis of MA in exfoliated cells. Conclusion MA was not associated with grade and stage of the bladder cancer. MA assay is a sensitive and effective method for the early detection of bladder cancer and post-operation surveillance.

Objective:To study the effect of Qingyi Lidan Granule on the serum levels of high mobility group box 1 (HMGB 1),heat shock protein 70 (HSP70),heat shock protein 27 (HSP27) and interleukin-8 (IL-8) of patients with severe acute pancreatitis.Methods:From August 2015 to July 2016,84 patients with severe acute pancreatitis in our hospital were selected and randomly divided into the observation group and the control group according to random number,42 cases in each group.The control group was given routine treatment,and the observation group was treated by Qingyi Lidan Granule on the basis of control group.The recovery of blood amylase to normal time,white blood cell recovery to normal time,recovery of gastrointestinal function to normal time and relieving time of abdominal pain,serum levels of HMGB1,HSP70,HSP72 and IL-8 in both groups were observed and compared before and after treatment.Results:After treatment,the total clinical effective rate of observation group was significantly higher than that of the control group[92.86％ (39/42) vs 71.43％ (30/42)] (P＜0.05).The recovery of blood amylase to normal time,white blood cell recovery to normal time,recovery of gastrointestinal function to normal time and relieving time of abdominal pain in the observation group were significantly shorter than those of the control group (P ＜0.05).Before treatment,no significant difference was found in the serum levels of HMGB 1,HSP70,HSP72,IL-8 between groups (P＞0.05).After treatment,the serum levels ofHMGB1,HSP70,HSP72 and IL-8 in both groups were significantly lower than those before treatment (P＜0.05).The levels ofHMGB1,HSP70,HSP72 and IL-8 in the observation group were lower than those in the control group (P＜0.05).Conclusion:Qingyi Lidan Granule could effectively reduce the levels of serum HMGB 1,H SP70,HSP27 and IL-8 and enhance the clinical curative effect of patients with severe acute pancreatitis.

AIM: To analyze the effect of human amniotic homogenate extract on corneal neovascularization after corneal alkali burn in the process of pigment epithelium derived factor (PEDF) and vascular endothelial growth factor (VEGF) expression and the effect of corneal neovascularization.METHODS: Totally 32 patients with corneal alkali burn were selected from June 2015 to June 2016 in Foshan,and were randomly divided into Group A and Group B,with a total of 37 eyes.Group A of 17 cases,with a total of 19 eyes,were treated with 40mg/L human amniotic homogenate extract;Group B (n=15),and 18 eyes,treated with 3g/L prednisolone eye drops.In the treatment of 1,4,7,14,21 and 28d at different time points,we observed the growth of corneal neovascularization,and detected the expression of PEDF and VEGF during angiogenesis.RESULTS: Group A of patients in the use of human amniotic homogenate extract after the treatment,the expression level of PEDF was significantly higher than that in Group B(P=0.001),after 28d treatment,the expression level of PEDF reached 0.721±0.314.While patients in Group B the expression level of PEDF was only 0.538±0.253.Two groups had significant difference between the expression level of PEDF (P<0.05).The expression level of VEGF in Group A was lower than in Group B at different time points in the test.After the treatment of 28d patients in the Group A,the expression level of VEGF was 0.152±0.020,in Group B the expression level of VEGF was0.302±0.031.Two groups of patients with VEGF expression level between the differences were statistically significant (P<0.05).The patients number in Group A with corneal neovascularization was significantly lower than that in Group B,the difference was statistically significant (P<0.05).CONCLUSION: Human amniotic homogenate extract can increase the expression of PEDF in corneal neovascularization after corneal alkali burn,inhibit the expression of VEGF and the proliferation of corneal neovascularization.

AIM: To investigate the inhibitory effect of quercetin on in vitro activation of T lymphocytes by polyclonal activators with CD69 expression as an activation marker. METHODS: After being separated from lymphoid nodes of a C57BL/6 mouse, the lymphocytes were exposed to polyclonal activators (PDB or Con A) with or without quercetin. Then they were harvested at 2 h, 6 h and 24 h, respectively. The expressional rates of CD69 on T lymphocytes were assessed by two-color immunofluorescent staining and flow cytometry, and the inhibitory rates of quercetin at different time points were estimated. RESULTS: Quercetin had no effect on the expressional rate of CD69 on T lymphocytes under resting states. After the stimulation with PDB or Con A, the expressional rates of CD69 on T lymphocytes in the present of quercetin (10 ?mol/L) showed significant decrease compared with those of control groups at different time points (P

A target molecule affinity-ultrafiltration liquid chromatography-electrospray ionization tandem mass spectrometric (LC-ESI-MSn) method was established for rapid screening acetylcholinesterase (AChE) inhibitors from total alkaloids of fibraurea recisa Pierre.A total of 12 potential inhibitors were screened from Fibraurea recisa Pierre.and 6 compounds were identified including palmatine,berberine,jatrorrhizine,palmatrubine,7,8-dihydro-8-hydroxyberberine and groenlandicine.The AChE inhibitory activity of these 6 compounds was validated in vitro.Palmatine showed the strongest inhibitory activity for AChE,which was stronger than that of donepezil hydrochloride,demonstrating the potential of palmatine as anti-Alzheimer's drug.This method is simple,rapid,and accurate for directly screening active ingredients which can inhibit AChE from complex extract of traditional Chinese medicines.

ABSTRACT:Objective To explore the effects of miRNA-1246 (miR-1246)on cell proliferation,invasion and migration in human cervical squamous cell carcinoma (CSCC)cell line SiHa.Methods SiHa cells were assigned into 3 groups:miR-1246 analog group,miR-1246 antagonist group and control group.Transfection efficiency was determined.The MTT assay,transwell assay and wound healing assay were performed respectively to evaluate the proliferation,invasion and migration abilities of SiHa cells.Western blot was carried out to detect the expression of thrombospondin-2 (THBS2)before and after transfection.A THBS2 3’-UTR-containing dual luciferase plasmid was synthesized and co-transfected with miR-1246 into SiHa cells to observe the luciferase enzyme activity.Results MTT assay,transwell assay and wound healing assay revealed that the abilities of proliferation,migration and invasion were significantly enhanced (P<0.01)in SiHa cells transfected with miR-1246 analog,but suppressed in SiHa cells transfected with miR-1246 antagonist.Western blot showed that SiHa cells transfected with miR-1246 analog had significantly decreased THBS2 expression (gray value = 6 .2 8 ± 1 0 .2 2 , P=0 .0 1 3 ) while those transfected with miR-1246 antagonist had significantly increased THBS2 expression (gray value = 12.90±19.81, P=0.037).After co-transfected with miR-1246 and THBS2 3’-UTR-containing plasmid,SiHa cells exhibited a decreased level of luciferase enzyme expression.Conclusion miR-1246 promoted the proliferation,invasion and migration of CSCC SiHa cell, and it might promote CSCC tumorigenesis and progression by suppressing the expression of its target gene THBS2 .

OBJECTIVE:To explore the protective effect and its mechanisms of Sinomenium acutum polysaccharide on mice with acute alcoholic liver injury. METHODS:60 mice were randomly divided into blank control group (normal saline),model group (normal saline),bifendate group (positive control,150 mg/kg) and S. acutum polysaccharide low-dose,medium-dose, high-dose groups(100,200,400 mg/kg),10 in each group,intragastrically administrated,once a day,for continual 7 d. 1 h af-ter last administration,mice received 50% ethanol (0.1 mL/10 g) intragastrically to induce acute alcoholic liver injury model ex-cept for those in blank control group. After 12 h,alanine aminotransferase(ALT),aspartate aminotransferase(AST)levels in se-rum,malondialdehyde(MDA),superoxide dismutase(SOD),glutathione(GSH),glutathione peroxidase(GSH-Px)levels in liv-er tissue of mice were determined;hematoxylin-eosin staining was conducted to observe the pathological changes in liver tissue;flow cytometry was used to detect the cell apoptosis rate. RESULTS:Compared with blank control group,mice in model group showed pathological changes in edema,disordered cell arrangement and local necrosis;ALT and AST levels in serum,MDA level in liver tissue and hepatocyte apoptotic rate were significantly increased,while the SOD,GSH and GSH-Px levels in liver tissue were significantly decreased,with statistical significances (P<0.01). Compared with model group,cell degeneration and necrosis degree of mice were improved in S. acutum polysaccharide medium-dose and high-dose groups;except for cell apoptosis rate of liver in S. acutum polysaccharide low-dose group was not decreased significantly,the above-mentioned indicators in other treatment groups were significantly improved(P<0.05 or P<0.01). CONCLUSIONS:S. acutum polysaccharide shows obvious protective effect on mice with acute alcoholic liver injury,its mechanism might re-late to anti-oxidation stress and inhibiting hepatocyte apoptosis.

In order to clarify the metabolism pathways of scandoside methyl ester, the analysis of metabolites profiling in four kinds of liver microsomes was performed by using an ultra-performance liquid chromatography/ electrospray-tandem mass spectrometry (UPLC-ESI-MS). The data obtained from the 0 h-incubation and the 2 h-incubation were compared and analyzed. After incubation, 5 metabolites of scandoside methyl ester were found in rat, Beagles, rhesus monkey and human liver microsome. The results showed that scandoside methyl ester's major metabolic pathway in the liver microsomes is hydrolysis, oxidation and reduction reactions, and there are certain kinds differences between species. The study provides a research base for further research about iridoid compounds in vivo metabolic pathways.

To identify the active components in Bufo melanostictus Schneider and clarify the difference between fresh and dried Venenum Bufonis, a UPLC-Orbitrap MS method has been established. The separation was performed with gradient elution of acetonitrile and water (with 0.1% formic acid) as mobile phase. By comparing their retention time and high resolution mass data of Venenum Bufonis extracts, 39 effective components were primarily identified by MS/MS analysis in positive ion mode. Twenty-six of them were bufadienolides. There were significant differences in the main composition between fresh and dried Venenum Bufonis. There are fewer bufadienolides in fresh toad venom.

Objective Discussed the efficacy of applying standardized patient in clinical surgical practice teaching.Method Twenty eight-year-medical students in our college were enrolled and students' theoretical knowledge mastering ability,students' evaluation on the teaching and students'clinical ability were surveyed.Result Students' score significantly increased compared to last year scores (P ＜0.01).100％ of the participants found the SP teaching methods can increase the interest in learning; 90％ of students found the SP as a guide to better grasp the teaching content; 95％ of students found the SP teaching methods can improve the ability to communicate with patients.Conclusions The class atmosphere was improved,students' enthusiasm was activated,students' clinical ability was promoted and satisfactory teaching effect was obtained after applying standardized patient.