OBJECTIVETo investigate the effects of sinomenine (SIN) and methotrexate (MTX) on the proliferation and apoptosis of in vitro cultured fibroblast-like synoviocytes (FLS) in rheumatoid arthritis (RA) patients, as well as the expression of osteoclast differentiation factor in FLS.

METHODSFLS were isolated from the synovium of RA patients and cultured in vitro. FLS were incubated with different concentrations of SIN and MTX respectively or combined: 0.001, 0.010, 0.100, 1.000 mg/mL SIN; 0.001, 0.010, 0.100, 1.000 mg/mL MTX; 0.001 mg/mL SIN + 0.001 mg/mL MTX, 0.010 mg/mL SIN + 0.010 mg/mL MTX, 0.100 mg/mL SIN + 0.100 mg/mL MTX, 1.000 mg/mL SIN + 1.000 mg/mL MTX, namely SIN1, 2, 3, 4 groups; MTX1, 2, 3, 4 groups and the combination 1, 2, 3, 4 groups. The medium without drugs was used as a control group. There was a total of 13 groups, each group with 3 complex holes. MTT was applied to detect the growth of FLS. The flow cytometry was applied to detect the apoptosis of FLS. The expressions of FLS receptor activator of nuclear factor kappa B ligand (RANKL) mRNA and osteoprotegerin (OPG) mRNA were observed by semi-quantitative RT-PCR.

RESULTSCompared with the control group, RA FLS proliferation OD values of all the drug groups were lower (P < 0.05). The RA FLS apoptosis OD value of the combination 3 group increased, the OPG mRNA expression increased, the expression of RANKL mRNA decreased with statistical difference (P < 0.05). The RA proliferation OD values of the SIN3 group and the MTX3 group increased when compared with the combination 3 group (P < 0.05).

CONCLUSIONSSIN and MTX had synergistic effects in inhibiting FLS. This might be one of the mechanisms for inhibiting RA bone damage.

Apoptosis ; drug effects ; Arthritis, Rheumatoid ; pathology ; Cell Proliferation ; drug effects ; Cells, Cultured ; Fibroblasts ; drug effects ; Humans ; Methotrexate ; pharmacology ; Morphinans ; pharmacology ; Synovial Membrane ; cytology ; drug effects

Objective To observe the change of invasion and migration of the pancreatic carcinoma cell line SW1990 transfeeted with EEF1A2 gene.Methods Pancreatic carcinoma cell line SW1990 was transfected with EEF1A2 by recombinant adenovirus vector.The alteration of motility、invasion and adhesion property of SW1990 was evaluated by wound healing assay,transwell With or without Matrigel basement membrane and adhesion assay.Results Wound healing assay revealed that EEF1A2 enhanced cell motility and transwell assay with Matrigel indicated that the average numbers of transwell cells with EEFlA2 was increased from 23.25±5.23 to 65.42±8.24(P＜0.05).The adhesive rate was substantially increased in EEF1A2 transfected SW1990 cells compared with control cells.Conclusions EEF1A2 gene can promote the migration.invasion and adhesion ability of pancreatic cancer cell in vitro.It is indicated that EEF1A2 may involve in the development of human pancreatic cancer by influencing cell biological characteristics.

Objective To investigate the effects of EEF1 A2 on growth and proliferation of the human pancreatic cancer cell line SW1990. MethodsThe EEF1 A2 gene was introduced into pancreatic cancer cell line SW1990 by adenovirus vector. The effects of EEF1 A2 on the growth of human pancreatic cancer cell were measured by MTT. Cell cycle was detected by flow cytometry and cell growth rate was examined by soft agar cloning formation test. ResultsAfter EEF1A2 induction, the expression of EEFA1 A 2 mRNA in pancreatic cancer cell line SW1990 increased, value of A750 at 72 h was 1. 2996 ±0. 2091, number of cells was 81250, cloning efficiency at 14 d was 82%, all of these parameters were significantly higher than those in the groups of blank adenoviras vector and PBS groups (P <0.05 ) ; the percentage of G1 phase cell was 28.5%, which was significantly lower than those in the groups of blank adenovirus vector and PBS groups; the percentage of Sphase ceil was 60.9%, which was significantly higher than those in the groups of blank adenovirus vector and PBS groups (P < 0.05 ). ConclusionsEEF1 A2 gene significantly enhanced cell growth and proliferation in human pancreatic cancer in vitro.

Participating in the National Oral Medical Skills Competition can push forward the teaching reform and curriculum construction,and promote the standardized development of teaching.According to the changes of competition level,rules and content of the National Oral Medical Skills Competition,the selection and training methods have been reformed.By designing the training programs,quantifying the scoring standards,strengthening the oral physician guidance and strengthening the psychological counseling and other trainings,satisfactory results have been achieved.

Humans ; Liver Cirrhosis, Biliary ; pathology ; surgery ; Liver Transplantation ; Recurrence

Objective To investigate the feasibility in screening of normal ovarian tissues by evaluating the expressions of tumor-associated genes in tissues adjacent to human epithelial ovarian cancer of orthotopic implantation in nude mice.Methods Human epithelial ovarian cancer cell lines OVCAR3 were grown in subcutaneous tissues,and the tumor tissues were orthotopic implanted.The expressions of CK-7,CA125,P53,survivin,MMP-2,and TIMP-2 were detected by immunohistochemical staining in proximal tissues,middle tissues,distal tissues adjacent to tumor,tumor tissues,and normal ovarian tissues of nude mice.Results 35 samples ovarian tissues with normal biopsy were gained from 40 cases of human epithelial ovarian cancers of orthotopic implantation model in nude mice.The expression rate of CK-7,CA125,P53,survivin,MMP-2,and TIMP-2 were 95.0％ (38/40),95.0％ (38/40),75.0％ (30/40),85.0％ (34/40),77.5％ (31/40),and 77.5％ (31/40) in tumor tissues,respectively; 71.4％ (25/35),68.6％(24/35),57.1％ (20/35),62.9％ (22/35),60.0％ (21/35),and 57.1％ (20/35) in proximal tissues adjacent to tumor,respectively; 34.3％ (12/35),31.4％ (11/35),31.4％ (11/35),31.4％ (11/35),34.3％ (12/35),and 31.4％ (11/35) in middle tissues adjacent to tumor,respectively; and 25.7％ (9/35),22.9％ (8/35),25.7％ (9/35),25.7％ (9/35),28.6％(10/35),and 31.4％ (11/35) in distal tissues adjacent to tumor,re respectively.The expressions of CK-7,CA125,P53,survivin,MMP-2,and TIMP-2 were all negative in 23 samples normal ovarian tissues adjacent to tumor.The expressions of CK-7,CA125,P53,survivin,MMP-2 and TIMP-2 in proximal tissues adjacent to tumor were lower than those in tumor tissues(P＜0.05),and higher than those in middle tissues and in distal tissues adjacent to tumor(P＜0.01).There were no expression difference between in middle tissues and in distal tissues (P＞0.05).The strong positive expressions of CK-7,CA125,and survivin were higher than those of P53,MMP-2,and TIMP-2 (P＜0.01).No significant difference was found in those expressions in the normal ovarian tissues adjacent to tumor gained from orthotopic implantation model with different severity.The expressions of CK-7,CA125,P53,survivin,MMP-2,and TIMP-2 were all negative in 20 normal ovarian tissues of nude mice.Conclusions The expression of CK-7,CA125,P53,survivin,MMP-2,and TIMP-2 showed decreasing trend to non-cancer direction.Negative expressions of these tumor-associated genes can be used as standard in screening of normal ovarian tissues adjacent to tumor.Relative safe normal ovarian tissues can be obtained from the tissues adjacent to tumor.

Objective To investigate the correlation between TCF7L2 gene rs3814570 polymorphisms with type 2 diabetes mellitus(T2DM) in Uygur population of Xinjiang area.Methods By adopting the case-control study design,949 cases of T2DM were recruited as the observation group and 963 individuals Undergoing healthy physical examination were selected as the control group.The TCF7L2 gene polymorphism was detected by matrix-assisted laser desorption/ionization-time of flight(MALDI-TOF).Results The statistical differences in frequencies of CC,CT and TT genotypes and the C and T allele frequencies on TCF7L2 rs3814570 were found between the T2DM group and control group(P＜0.05).The risk of suffering from T2DM in the carriers of CT genotype was 0.331 times of that in the carriers of CC genotype(OR =0.331,95 ％ CI:0.166-0.661,P =0.002),the risk of suffering from T2DM in the carriers of TT genotype was 0.539 times of that in the carriers of CC genotype(OR=0.539,95％CI:0.348-0.834,P=0.005),and the risk of suffering from T2DM in the carriers of T allele was 0.501 times of that in the carriers of C allele(OR=0.501,95 ％ CI:0.377-0.664,P＜ 0.01).Among all subjects,the FPG level of the CT + TT genotype group on TCF7L2 gene rs3814570 locus was significantly lower than that of the CC genotype group(P＜0.05).Conclusion The rs3814570 locus in TCF7L2 gene may be associated with T2DM occurrence in Uygur population of Xinjiang area,the T allele and TT genotype might be protective factors of T2DM.

OBJECTIVETo observe the clinical efficacy of ursodeoxycholic acid (UDCA) and Fuzheng Huayu Capsule (FHC) in the treatment of primary biliary cirrhosis (PBC).

METHODSEighty PBC patients were randomly assigned to two groups, the treatment group and the control group, 40 in each group. Patients in the treatment group took UDCA and FHC, while those in the control group were treated with UDCA alone. The treatment course was 48 weeks for both groups. The clinical symptoms and signs, liver function indices (ALT, AST, ALP, GGT, ALB, TBIL, and TBA), hepatic fibrosis indices (HA, LN, IV-CL, and PIIIP), immunologic indices (IgG, IgM, and autoimmune antibodies), changes of portal hemodynamics, and adverse reactions were observed before treatment, as well as at week 4, 12, 24, and 48 after treatment.

RESULTSAfter treatment the skin itching and fatigue were significantly improved in the treatment group, showing statistical difference when compared with the control group (P < 0.05, P < 0.01). After treatment the levels of ALT, AST, ALP, GGT, TBIL, and TBA obviously decreased in the two groups. They were lower in the treatment group than in the control group at the same time point (P < 0.05). The decrement was the largest at week 4. Besides, at week 48 after treatment the ALB level was improved in the treatment group (P < 0.05). The levels of HA and PIIIP obviously decreased at week 4, 12, and 24, the levels of LN and IV-C obviously decreased at week 4 and 12, the decrement of the hepatic fibrosis indices at week 4 were more obvious in the treatment group. But the levels of HA and PIIIP were lower than the pre-treatment levels at week 12 in the control group. The immunologic indices such as IgM and IgG were improved in the two groups, with better results obtained in the treatment group (P < 0.05, P < 0.01). In the treatment group ANA turned negative in 1 patient and AMA turned negative in 2 patients. After 48 weeks of treatment, the spleen was retracted, the inner diameters of the portal vein (PV) and the splenic vein (SV) were significantly reduced, and the blood flow velocity in the PV and SV increased in the treatment group (P < 0.01). At week 24 and 48, 33 patients (82.5%) and 26 patients (90.0%) in the treatment group had complete relief, better than those of the control group [22 cases (55.0%) and 28 cases (70.0%)]. No obvious adverse reaction was found in the two groups during the treatment course.

CONCLUSIONSThe combination therapy of UDCA and FHC was effective and safe in anti fibrosis and improving the liver functions of PBC patients. It was safe and better than the application of UDCA alone. It was advocated to be combined use for a long term. It might improve the long-term efficacy.

Adult ; Capsules ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Liver Cirrhosis, Biliary ; drug therapy ; Male ; Middle Aged ; Phytotherapy ; Ursodeoxycholic Acid ; therapeutic use ; Young Adult

Objective To detect anti-M_2 autoantibody using recombinant BCOADC-E_2.Methods We purified recombinant BCOADC-E_2 by Ni~(2+)affinity chromatography column and then detect anti-M_2 autoan- tibody in the sera of patients with primary biliary cirrhosis(PBC)by Western blot test(WBT)and enzyme linked immunosorbent assays(ELISA).Results Among 60 sera from PBC patients,33 were positive,all of controls were negative.Conclusion The recombinant BCOADC-E_2 can be used to detect anti-M_2 autoanti- body specifically and sensitively.It is helpful for the diagnosis of PBC.

OBJECTIVETo study the effects of Zhengqing Fengtongning Tablet (ZFT) and methotrexate (MTX) on the expression of osteoprotegerin (OPG), receptor activator of nuclear factor-kappaB ligand (RANKL), and interleukin 17 (IL-17) in collagen-induced arthritis (CIA) rats, thus addressing their bone protection.

METHODSThe CIA rat model was established by intradermally injecting type II collagen emulsion from the rats' back and tail. Totally 28 successfully modeled rats [with the arthritis index (AI) more than 2] were randomly divided into the model group, the Chinese medicine (CM) treatment group, the MTX group, and the ZFT + MTX treatment group, 7 rats in each group. Another 7 rats were recruited as the normal control group. Rats were administered from the 7th day of modeling. Rats in the MTX group were treated with MTX at 3.8 mg/kg once a week. Those in the CM group were treated with ZFT at the daily dose of 130 mg/kg, once a day. Those in the ZFT + MTX treatment group were treated with both MTX (at 3.8 mg/kg once a week) and ZFT (at the daily dose of 130 mg/kg, once a day). Those in the model group and the normal control group were administered with normal saline of the equal volume by gastrogavage. All the intervention lasted for 26 days. The destruction of joints in the four limbs were observed using X-ray. The AI was recorded. The expression levels of serum OPG, RANKL, and IL-17 were detected at the end of the experiment.

RESULTSDuring the whole process, all rats except those in the model group were in a good condition. On the 21st day of modeling the AI of all rats reached the peak, but it decreased after treatment. Compared with the model group, the AI decreased in the CM treatment group, the MTX group, and the ZFT + MTX treatment group with statistical difference (P < 0.05). Compared with the model group, the OPG increased and RANKL decreased in the MTX group; the OPG and OPG/RANKL increased in the CM treatment group; the OPG, RANKL, and OPG/RANKL increased, and IL-17 decreased in the ZFT + MTX treatment group, all showing statistical difference (P < 0.05). Compared with the MTX and the ZFT + MTX treatment group, OPG/RANKL increased and IL-17 decreased in the ZFT + MTX treatment group (both P < 0.05).

CONCLUSIONZFT + MTX could synergistically elevate peripheral OPG/RANKL and down-regulate IL-17 in CIA model rats.

Animals ; Arthritis, Experimental ; blood ; chemically induced ; drug therapy ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Interleukin-17 ; blood ; Methotrexate ; pharmacology ; therapeutic use ; Osteoprotegerin ; blood ; RANK Ligand ; blood ; Rats