Tumor necrosis factor alpha (TNF-α) is a homotrimeric protein encoded within the major histocompatibility complex (MHC). TNF-α can bind its receptors in the body to play a role in immune regulation and to participate in the various pathophysiology processes including fever, inflammation, infection, wound healing and tumors necrosis. TNF-α, an important component of inflammatory pathways, is up-regulated in the synovial tissue, synovial fluid and serum of rheumatoid arthritis (RA) patients. In RA patients, TNF-α along with many kinds of tissue factors and matrix proteins together promotes the inflammatory response, abnormal apoptosis of synovial cells, pannus formation and cartilage and bone destruction, which maintain the sustainability of the progress of RA. Thus, TNF-α may be a clinical indicator of RA activity as well as an effective target for RA.

Liquichip is a new type of biochip emerging in the 1990s, which is integrated flow cytometry, laser technology, digital signal processing and traditional chemical technology with the greatest features of high-throughput and high-flexibility. Luminex has promoted new products from Luminex 100/200 to Flexmap 3D. This is an introduction to the development, principle, and application of Luminex liquichip.

Objective To study the effects of cripto on migration, invasion, and liver metastasis of colorectal cancer cell. Methods After human colorectal cancer cell line SW480 was transfected by cripto small interfering RNA (siRNA), the mRNA and protein level were determined by Real-time RT-PCR and Western blot, respectively. The migration and invasion ability were evaluated by wound-healing assay and boyden chamber model, respectively. Thirty nude mice model of liver metastasis from colorectal cancer was established by splenectomy. Results The siRNA could down-regulate the level of mRNA and protein of cripto in a dose- and time-dependent manner. Suppression of cripto expression could inhibit migration and invasion ability of human colorectal cancer cell in vitro. The metastastic rate and tumor nodules were lower in transfection with cripto siRNA than in two control groups in vivo. Conclusions Cripto gene might play an important role in regulation of liver metastasis from colorectal carcinoma cell, and suppression of cripto gene by siRNA can inhibit liver metastasis of colorectal cancer.

Objective To introduce dimethyl on 4-position of 4-ene-3-one steroids. Methods Seven 4-ene-3-one steroids were treated with methyl iodide (CH3I) and potassium tert-butoxide (t-BuOK) in tert-butanol (t-BuOH). Results Four substrates were smoothly converted to 4,4-dimethyl products, while other three substrates afforded 3-O-methylation products with high yields. Conclusions The different reaction results revealed a high remote effect of D ring moiety on the selectivity between 3-O-methylation and 4-dimethylation.

Objective To prepare monoclonal antibody in mice so as to develop an ELISA method for diagnosis of Toxoplasma gondii infection during the initial stage. Methods The mice were immunized by combining routine and intrasplenic immunization with recombinant SGA1 antigen. B lymphocyte hybridization technique was applied to prepare the anti-SAG1 McAbs. Positive clones were screened using ELISA and subcloned to establish cell lines. Ascites was induced to produce the McAbs. Then the McAbs were purified by protein G chromatograph column. The specificity of McAbs was identified by Western blot and sandwich-ELISA. Sensitivity of the McAbs was determined using sandwich-ELISA. Comparasion was carried out between PCR and sandwich-ELISA method. Results Two positive clones were obtained and named as 3B6, 10C4, both could identify the native and recombinant SAG1 antigens. The sensitivity of 3B6, 10C4 was 31.3 ng and 62.5 ng, respectively. There was no cross reaction between the McAbs and positive sera from patients with schistosomiasis, ancylostomiasis or malaria. By using PCR and ELISA, the positive infection rate of T. gondii was 63.2% and 47.4%, respectively. Conclusions Therefore, mouse anti-rSAG1 antigen McAbs have been prepared successfully and primarily applied to early stage diagnosis of T. gondii infection.

Objective To investigate the inhibitory effect and the mechanism of endostatin on human gastric carcinoma model in nude mice. Methods Athymic mice were injected by in vitro-cultured SGC-7901 human carcinoma cell strain to observe the inhibitory effect of endostatin. Real time fluorescent quantitative PCR and immunohistochemistry stain were used to quantify the expression levels of Bcl-2 and Bax genes both on mRNA and protein levels. Results The average tumor volume in endostatin group was statistically smaller than control group (P<0.05). The inhibitory rate on the 7~(th), 10~(th), 13~(th), 16~(th) and 19th day was 26.2%, 47.7%, 38.9%, 45.8% and 51.7%, respectively. The results of real time fluorescent quantitative PCR demonstrated that mRNA expression of Bcl-2 in endostatin group was obviously lower than it in control group (P<0.05), however, the expression level of Bax had no statistical difference between these two groups (P>0.05). The results of immonohistochemistry were consistent with PCR. Conclusions Endostatin can inhibit the proliferation of SGC-7901 human gastric carcinoma in athymic mouse model, and the mechanism involves the suppression of Bcl-2 expression.

Objective To investigate the effect of Schistosoma japonicum ova on the expression of intestinal NOD2/CARD15 in the mice induced by 2,4,6-trinitrobenzesulfonic acid (TNBS). Methods Mice (n=50) in the experiment were randomly allocated into 3 groups: control group (n=10), TNBS+saline (n=20) and TNBS+Schistosoma japonicum ova (n=20). TNBS enema (100 mg/kg) was applied to the two TNBS groups in order to establish a colonitis model. Schistosoma japonicum ova was administered i.p. on the 14~(th) and 3~(rd) day before the instillation of haptenating agent. All mice were killed on the 7~(th) day after colitis induction. The transcription level of NOD2 in colon tissues was measured by Real time PCR, and the expression of NOD2 protein was measured by Western blot. Results The transcription and protein levels of NOD2 in TNBS-induced mice increased statistically compared with those of the normal group (P<0.05). Compared with the TNBS-induced mice, Schistosoma japonicum ova-treated ones exhibited a statistical reduction of gene and protein expression (P<0.05). Conclusions TNBS-treated mice exhibited a statistical increased expression of NOD2/CARD15, Schistosoma japonicum ova treatment reduced the severity of experimental colitis through down-regulating NOD2/CARD15.

Objective To construct the recombinant expression vector encoding antisense Tcf fragment for the blockage of abnormal Wnt pathway, and to investigate its effect on the biological behaviors of human hepatocarcinoma cells. Methods Antisense expression vector was transfected into hepatocarcinoma cells SMMC-7721 with GeneJammer. RT-PCR and Western blot were used to detect Tcf expression. Cell proliferation and motility were compared by growth curves and Transwell plate assay. Cell apoptosis was determined by Annexin V and cell cycle was examined by fluorescent staining. Results The stable transfection of antisense Tcf in SMMC-7721 cells significantly reduced Tcf expression at both mRNA and protein levels. Compared with parental and mock-transfected 7721 (7721-vector) cells, antisense Tcf RNA transfected cells 7721-pTas showed much decreased activities of proliferation, migration and invasion in vitro. Furthermore, the apoptosis rate of 7721-pTas cells [(26.34±2.07)%] was significantly higher than that of 7721-vector cells [(6.53±1.02)%] and parental SMMC-7721 cells [(4.33±0.68)%] (P<0.001). The percentages of G0-G1 phase antisense transfected cells were 20.24% and 20.95%, higher than parental SMMC-7721 and 7721-vector cells, and percentages of S phase antisense transfected cells were 11.8% and 11.38%, lower than parental SMMC-7721 and 7721-vector cells, respectively. Conclusions Antisense RNA suppress the growth ability of liver cancer cells by inducing cell apoptosis and impeding the progress of cell cycle, which suggests that selective blockage of abnormal Wnt signal pathway by antisense Tcf RNA may be a potential new gene therapy for liver cancer.

Objective To investigate the impact of aging and weight bearing on cartilage endplate morphology and chondrocyte apoptosis in rats. Methods The bipedal rat model (n=45) was developed by forelimb amputation and special breeding methods. The normal rats of the same age served as the control group (n=40). When the rats became 3, 6, 9, and 12 months old, 8 rats randomly selected from each group were sacrificed and paraffin-embedded mid-sagittal sections of the L4-5 spine were obtained. Sections were stained with hematoxylin and eosin and the TUNEL procedure was performed. The numbers of apoptotic cells and viable cells in the cartilage endplates of the intervertebral discs were counted, the thickness of the cartilage endplate was measured and the degree of impairment of the cartilage endplate was evaluated. Results Apoptosis first appeared in the cartilage endplate, then increased with aging and resulted in a remarkable decrease in cell density. The apoptotic rate of chondrocytes within the cartilage endplate of the bipedal rat model group was significantly higher than the control group at the 6-month time point. A statistically significant difference was observed in the bipedal rat model group between the 6-month time point and 9-month time point (P<0.05). Correlation analyses indicated that there was a highly negative correlation between the number of the viable cells of the cartilage endplate and the degree of the cartilage endplate degeneration (r=-0.97, P<0.05). Compared with the naturally aged group, the bipedal rat model group experienced more severe degeneration in the structure of the cartilage endplate, more obvious thickening of the cartilage endplate's calcified layer, and more defects in the structure of the cartilage. Conclusions Besides aging, weight bearing is probably a key factor of the increase of chondrocyte apoptosis and the degeneration of vertebral cartilage endplate.

Objective To assess the efficacy of ganciclovir to prevent and cure cytomegalovirus (CMV) infection after renal transplantation. Methods We searched PubMed, EMBASE, Cochrane Library, SCI, China Academic Journals Full-text Databases, Chinese Biomedical Literature Database, Chinese Scientific Journals Databases and Chinese Medical Association Journals to collect randomized controlled trials of ganciclovir to prevent and cure CMV infection after renal transplantation (up to June, 2009). Two reviewers extracted data independently using a designed extraction form. The quality of included trials was evaluated according to the Cochrane Handbook. RevMan 5.0 software was used for data analysis. Results Twelve randomized controlled trials were included. The results of meta-analysis showed that: ①Compared with no receive antiviral agents, ganciclovir couldn't lower CMV infection rate and disease rate in 3 months and 6 months after renal transplantation, but could lower CMV disease rate in 12 months. The delay between transplantation and CMV infection was significantly longer. ②Either valaciclovir or ganciclovir could lower CMV infection rate and disease rate after renal transplantation, without statistical difference. ③Compared with acyclovir, ganciclovir could lower CMV disease rate in 6 months after renal transplantation. ④Compared with CMV-IgG and valganciclovir, ganciclovir didn't have statistical difference in decreasing CMV disease rate (P=0.93;P=0.14). Conclusions Longer prophylaxis by ganciclovior may prevent CMV infection after renal transplantation. Its curative effect is similar to valaciclovir, CMV-IgG and valganciclovir, but better than acyclovir.