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 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.

Abstract: Objective To research the relationship between promoter polymorphism-438 A/G of thrombin-activatable fibrinolysis inhibitorgene (TAFI-438 A/G) and atherosclerotic cerebral infarction (ACI) in Chinese Han population. Methods TAFI-438A/G genotypes andtheir allele frequencies were identified with the polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) in225 ACI patients and 184 healthy controls. Results There was no significant difference of TAFI-438A/G polymorphism between ACI groupand control group. Stratified by gender, in males, the incidence of A allele was 28.6% in ACI group, and 20.6% in control (P=0.039); of theAA genotype was 9.0% in ACI group, and 1.9% in control (P=0.019). But no significant difference was found in females. ConclusionTAFI-438A/G polymorphism is associated with the risk of ACI in males, that AA genotype may increase the risk of ACI.

Objective To explore the role of sialic acid-binding immunoglobulin-like lectin-one (Siglec-1) in the process of atherosclerotic inflammation induced by oxidized low-density hpepmtein (ox-LDL). Methods Ox-LDL was synthesized by oxidization of native LDL and different concentration of ox-LDL was added to the culture medium of RAW264.7. Forty-eight hours later, cells and supernatants were collected separately. The expression of Siglec-1 protein and mRNA were measured by flow cytometry(FCM) and real-time quantitative RT-PCR, respectively. The levels of monocyte chemotactic protein-1 (MCP-1), macrophage inflammatory protein-1α(MIP-1α) and IL-8 in supernatants were determined by ELISA. Re-sults By the stimulation of ox-LDL, Siglec-1 protein and mRNA on RAW264.7 cells were significantly in-creased, meanwhile, the cytokines levels in culture supematants were significantly higher than that in the control group. And both Siglec-1 expression and cytokine secretion were ox-LDL dose-dependent. Conclu-sion Ox-LDL can increase Siglec-1 protein and mRNA expression and some inflammatory cytokines secre-tion on RAW264.7 cells in a dose-dependent manner. Manifested by enhanced Siglec-1 expression, the acti-vated macmphages may take a part in the development and progression of atherosclerosis.

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 study the immunoregulatory effects of thalidomide on the peripheral blood T-lymphocytes of rheumatoid arthritis patients.Methods MTr was used to detect the effects of different thalidomide concentrations on the proliferation of T-cells.Flow eytometry was used to analyze T-cells early apoptosis and the T-cells subsets in different concentration of thalidomide.The mRNA expression of IL-6,IL- 10 and TNF-α was measured by RT-PCR method.Results The level of thalidomide at 500 μg/ml inhibited the proliferation of T-ceils and the CD3+CD28+ expression of T-cell subsets,but promoted the early apoptosis and the CD8+CD152+ expression of T-cell subsets.Thalidomine at any concentration could inhibit the mRNA expression of IL-6,TNF-α.However,the level of thalidomide that could promote the mRNA expression of IL- 10 was 100 μg/ml and 500 μg/ml.Conclusion Thalidomide can inhibit the proliferation of T lymphocytes and the expression of CD3+CD28+ on T-cell subsets.It can promote the early apoptosis and the CD8+CD152+ expression of T-cell subsets.Thalidomide inhibits the mRNA expression of IL-6 and TNF-α but promote the mRNA expression of IL-10.Thalidomide has immuno-regulatory effects on rheumatoid arthritis T-cells.

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 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 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.