Objective To establish the extraction process of Zhechong Chuangyu Capsule. Methods The difference of analgesic effect of water extraction and alcohol extraction in mice was observed by body-torsion test to determine the extract solvent. With the rate of aqueous extraction, n-butyl alcohol extraction and asperosaponin Ⅵ as evaluating indicator, the influencing factors including solvent volume, time and times of extraction were investegated to evaluate extracting procedure by orthogonal experiment. Results There was no obvious difference in analgesic effect between water extraction and alcohol extraction. Given the requirement of produce, aqueous extraction was a better choice. The optimum extracting condition was extracted 3 times with 20 folds volume of solvent, and extraction time was 150 minutes. Conclusion The extraction process is feasible to be applied into production.

Objective To investigate the clinical features and prognosis of patients with de novo digestive system malignancy after liver transplantation.Methods The clinical data of 10 patients with de novo digestive system malignancy out of 1 517 patients undergoing liver transplantation from January 2004 to December 2015 in the First Affiliated Hospital of Zhejiang University were collected.Immunosuppressive agents,type of de novo malignancy,process of treatment and prognosis were analyzed retrospectively.Results The incidence of de novo digestive system malignancy after liver transplantation was 0.7％ (10/1 517),including 3 cases of gastric cancer,3 cases of colon cancer,2 cases of liver cancer,2 cases of pancreatic cancer.All of the 10 patients received the corresponding treatment,and the dosage of immunosuppressive agents was reduced at the same time.The median time from liver transplantation to the diagnosis of de novo malignancy was 40.5 months (23 to 156 months).The median follow-up time was 10.5 months (3 to 61 months) after they had been diagnosed as malignancies,and 6 patients died of tumor progression.Conclusions The prognosis of patients with de novo digestive system malignancy after liver transplantation is poor,and early diagnosis and treatment is an important way to improve prognosis.Liver transplant recipients need to be closely followed up,especially in patients with high risk factors of digestive system malignancy.The modification of immunosuppressive drugs may be an effective way to prevent and improve prognosis.

Objective To explore the association of ubiquitin-specific proteases 24 (USP24) gene polymorphisms with susceptibility to sporadic Parkinson's disease (PD) in the Han Guangdong population. Methods From August, 2006 to January, 2014, single nucleotide poly-morphisms (SNPs) of rs12138592 and rs6671533 in the intron region of USP24 were genotyped in 200 patients with sporadic PD and 200 healthy controls using the SNaPshot technique. Results There was significant difference in the allele and genotype frequency of rs12138592 between the patients and the controls (P<0.01), and no significant difference was found in the allele and genotype frequency of rs6671533 (P>0.05). Conclusion The SNP of rs12138592 in the intron region of USP24 is associated with the susceptibility to sporadic PD in the Han Guangdong population, and the A allele may contribute a protective roles to PD.

Objective To study the efficacy of emergency orthotopic liver transplantation(EOLT) for acute(hepatic) failure(ALT).Methods A retrospective review was undertaken on the clinical data of 8 patients undergoing emergency liver transplantation for ALT.Results The 8 patients completely regained consciousness in 12 to 72 hours after operation.No case developed central nervous complications.One case of severe(hepatitis) complicated by acute renal failure died of respiratory infection and ARDS on postoperative day 7.One case who refused to take medication died from chronic rejection 12 months after operation.One case was(complicated) by bile duct stricture and biliary sludge at 14 months postoperatively and survived for 18 months.Four of the other 5 cases were followed up for 17 months and 1 cases for 14 months,and thir quality of life was excellent.3 of them have returned to work.Conclusions Emergency orthotopic liver thansplantation is an effective means to treat ALF.Intensive care and effective treatment preoperatively are pre-requisite(conditions) to ensure the success of EOLT.

Objective To study the important virulence regulation genes of Brucella,and to understand their function.Methods Quantitative RT-PCR was used to quantify their relative transcription profiles under stress conditions and during macrophage cell infection.Results These genes were activated at different levels under these conditions and during cell infection,indicating their roles in pathogenesis at different srage of infection.Conclusion The transcription profiles of these genes have different effects about their functions.

Objective To investigate the increased expression of microRNA-146a(miR-146a) in peripheral blood mononuclear cells (PBMC) of patients with chronic immune thrombocytopenic purpura (ITP) and its clinical significance. Methods Twenty-eight patients with chronic ITP and 28 healthy controls matched with age and gender were enrolled in this study. Fluorescent quantitative PCR reaction was used to detect the relative expression of miR-146a in their PBMC. The serum concentration of TNF-α, IL-2,IL-1 β and IFN-γ were measured by ELISA. CCK-8 method was used to detect the proliferation ability of PBMC , which transfected with miR-146a mimics or inhibitor and then stimulated with platelet . Results The relative expression of miR-146a in ITP patients was higher than that of healthy controls. The increased expression of miR-146a was negatively correlated with the serum TNF-α, IL-2 and IFN-γ. The PBMC transfected with miR-146a mimics had reduced expression of IL-2 and proliferation when stimulated with platelet.In contrast, the opposite effect was observed with the miR-146a inhibitors transfection. Conclusion MiR146a was involved in the pathogenesis of chronic ITP by controlling IL-2 production and PBMC proliferation.Thus, it may be a potential therapy target for chronic ITP.

Objective To detect the expression of IL-27 in PBC (primary biliary cirrhosis)patients and the possible involvement of IL-27 signal pathway in PBC. Methods The gene transcription and protein expression levels of IL-27 in patients with PBC, chronic hepatitis B(CHB) group and healthy controls(HC) were measured by real-time PCR, ELISA, flow cytometry and immunohistochemistry. AST,ALP, ALT, TBIL, GGT were determined and their correlation with IL-27 was also analyzed. Results IL-27 was significantly elevated in patients with PBC and IL-27 is present in the liver tissues of patients with PBC. Expression of IL-27 on CD4+T cells was increased in patients with PBC(72.40% ±6.22% ) and CHB(59.40% ± 7.03%) compared with HC(1.70%±0.55%,P＜0.01). Expression of IL-27 protein was increased in patients with PBC [( 126.25 ± 36.00 ) pg/ml] compared with CHB [( 51.81 ± 23.30 )pg/ml, P ＜ 0. 01] and HC[(34.19 ± 9.70) pg/ml, P ＜ 0.01], and it was positively correlated with GGT( r = 0.554, P＜0.01) and TBIL (r = 0.559,P＜0.01), but no correlation with ALT, AST, ALP.Conclusion These facts indicated the key role of IL-27 in the immune inflammatory reaction in patients with PBC.

Objective To investigate the imaging findings and key diagnostic points of Brunner’s gland adenoma on CT.Methods The CT imaging findings and pathological features of 9 cases of Brunner’s gland adenoma confirmed by pathology were retrospec-tively analyzed,including the lesions number,site,size,shape,margin,density and the enhancement pattern.Results Of the 9 ca-ses,1 case located in the antrum and 8 cases in the duodenaum [6 cases in the duodenal bulb (75%)and 2 in the papillary (25%)]. Of the 6 cases of duodenal lesions,3 were found at the anterior wall and 3 at the posterior wall.Except 1 case which complicated with enteritis and had an obscure margin,the other 8 cases were clear margined ,and were round or nodular in shape.The maximum size of tumors ranged 1 5-68 mm in diameters (mean 35.0 mm ± 1 6.2 mm).The density of tumors was homogeneous on CT scan without necrosis or hemorrhage.In the arterial phase after administration of contrast agent,the lesions were similar to the adjacent intestinal wall enhancement,and mucosal annular enhancement (halo sign)showed in 6 cases,and the dot-shape non-enhancement area within the lesion (black star sign)showed in 5 cases,and the thickening or tortuous enhanced blood vessel showed in 6 cases.In the venous phase,9 cases were progressive enhancement,and the “black star sign”or “the black line sign”showed more clearly in 5 cases.In the pathology,the lesions were polypoid-like,solid or cystic.Under the microscope,the hyperplasic Brunner’s glands were covered with normal duodenal mucosa and separated by bundles of smooth muscle cells with dilated duct,cyst,and adipose cells,1 case with atypical hyperplasia of the glandular epithelium and 1 case with ectopic pancreas.Conclusion There are some spe-cific CT imaging features in Brunner’s gland adenoma,which is of important clinical value in accurate preoperative diagnostic.

Objective To investigate the effect of miR-146a on the proliferation and interleukin (IL)-2 production of T helper cells from primary biliary cirrhosis (PBC) patients.Methods MiR-146a in the peripheral blood mononuclear cells (PBMC),monocytes,T helper cells,cytotoxic T cells and B cells from 20 confirmede PBC patients and age/sex matched healthy controls were detected by quantitative PCR.By gainand-loss of function,the miR-146a's effect on anti-CD3/anti-CD28 activated T helper's proliferation and IL-2 production ability were measured by CCK-8 approach and enzyme linked immunosorbent assays (ELISA),respectively.Statistical analysis were carried out by t-test.Results PBMCs (0.46±0.20 vs 1.00±0.26; t=7.47,P＜0.01),T helpers (0.33±0.13 vs 1.00±0.14; t=6.15,P＜0.01) and monocytes (0.56±0.11 vs 1.00±0.11; t=4.97,P＜0.05),but not B cells (0.91±0.06 vs 1.00±0.14; t=0.97,P＞0.05) and cytotoxic T cells (0.98±0.15vs 1.00±0.12; t=0.22; P＞0.05) from PBC patients had lower miR-146a expression level than that of healthy controls.Inducible up expression of miR-146a was observed in PBC patients'T helpers stimulated with antiCD3/anti-CD28 (1.00±0.18 vs 9.12±2.05; t=8.81; P＜0.01).The activated T helpers from PBC patients had higher proliferative ability [PBC:0.35±0.06 (A); healthy controls:0.26±0.04 (A); t=2.83; P＜0.05] and increased IL-2 production [PBC: (685.60±109.19 pg/ml)]; Healthy controls: [(512.20±72.26) pg/ml; t=2.96; P＜0.05 ] than those of healthy controls.For activated T helpers,the proliferation ability,as well as IL-2 production,was enhanced by miR-146a.Conclusion MiR-146a can down regulate the proliferation and IL-2 releasing of activated T helpers.The reduced miR-146a expression enhances IL-2 production and promotes proliferation of T helper of PBC patients,thus,may be involved in the pathogenesis of PBC.

ObjectiveTo investigate mechanisms for IL-27 induced proliferation and differentiation of peripheral blood CD4+ T cells in primary biliary cirrhosis (PBC).MethodsPeriperal blood CD4+ T cells were isolated from patients with PBC,chonic hepatitis B (CHB) and health controls (HCs).After IL-27 stimulation,proliferation ability of CD4+ T cells was evaluated by CCK-8 kit,and cytokines were analyzed by ELISA.Real-time PCR was employed to assay mRNA expression of T-bet and GATA3 in CD4+ T cells.p-STAT-1 and pSTAT-3 expression in CD4+ T cells were detected by Western blot.ResultsEnhanced proliferation of CD4+ T cells was found in all subjects after IL-27 stimulation.However,the proliferation ability in patients with PBC was greater than that in CHB and HCs ( P＜0.001 ).Levels of IL-2 and IFN-γ in supernatant from IL-27-incubated PBC blood CD4+ T cells were higher than that from CHB and HCs (P＜0.001 ).In normal situation,T-bet mRNA of CD4+ T cells in PBC group was higher than that in CHB group (P=0.007).Furthermore,after IL-27 stimulation,elevated T-bet mRNA expression and GATA3 inhibition were found in patients with PBC.High expression of p-STAT-1 and p-STAT-3 in blood CD4+ T cells were found in PBC,CHB and HCs after stimulation by IL-27.But their expression in patients with PBC were higher than those in patients with CHB and HCs.ConclusionProliferation of blood CD4+ T cells could be induced by IL-27 in patients with PBC.The signaling pathways of p-STAT-1,p-STAT-3 were involved to induce Th1 immune response and related cytokines expression.This study implicated that IL-27 may play important roles in early inflammation damage in PBC.