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.

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.

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 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 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 understand the screening results and positive lesion situation of colonoscopy screening among high risk urban population of colon cancer in Chongqing Nan'an District to provide a basis for the community heath guidance.Methods The high risk subjects of colon cancer screened according to the item of the Early Diagnosis and Early Treatment of China Urban Cancer and in our center for completing colonoscopy screening during 2013-2016 were collected and analyzed on the positive rates of various lesions.Results A total of 435 subjects were included,141 positive cases were found with the total positive rate of 32.41％ (141/435),in which 75 cases were male positive with the positive rate of 47.47％ (75/158),and 66 cases were female positive with the positive rate of 23.83 ％ (66/277),the polyps detective rate was 28.27 ％ (123/435),tumor-like lesion detective rate was 7.59％(33/435),the adenocarcinoma detection rate was 0.46％ (2/435);the total positive rate of male was higher than that of female(x2 =25.669,P＜0.01).Conclusion The sequential screening mode in which the high risk population preliminarily screened out by community then performs colonoscopy examination has an important significance for discovering the intestinal tract lesion.

In this study ,we established Cross Priming Amplification (CPA ) technology for detection of influenza A virus (H1N1) approach ,and evaluated the method through clinical specimens .A set of specific primers were designed for CPA ac‐cording to the conservative gene sequences ,designed and realized in the same temperature reverse transcription of RNA and DNA amplification . The amplification products can be totally enclosed nucleic acid detection device for testing . Fourteen healthy pharyngeal swab specimens ,seven other respiratory viruses ,and six arboviruses strains were used as the controls .We used a method that application of gradient dilution to the H 1N1 virus strain as the control to test the sensitivity of the CPA .We also used 102 clinical pharyngeal swab specimens of H1N1 patients for detection object to evaluate the feasibility of CPA clinical detection .Results showed that the CPA reaction did not appear cross reaction on health cases samples and other viruses .The sensitivity of the CPA was approximately 10 copies/uL in the established method that exactly titer H1N1 virus strain gradient dilution test .As to the positive results among the clinical pharyngeal swab samples collected from patients at different stages after onset ,the CPA had the highest positive detection rate during the first three days after onset (100% ) .While the detection rate from day 4 to day 6 after onset was 79 .31% .After 7 days ,the detection rate was 9 .09% .The established CPA assay was a highly sensitive ,specific and reproducible approach for rapid detection of H1N1 virus ,which is conducive to the early diagno‐sis of influenza A virus (H1N1) for basic medical units .

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.