Gene expression profiling defined pathways correlated with fibroblast cell proliferation induced by Opisthorchis viverrini excretory /secretory product

AIM： To investigate the mechanism of fibroblast cell proliferation stimulated by the Opisthorchis viverrini excretory/secretory （ES） product. METHODS： NIH-3T3, mouse fibroblast cells were treated with O. viverrini ES product by non-contact co-cultured with the adult parasites. Total RNA from NIH-3T3 treated and untreated with O. viverrini was extracted, reverse transcribed and hybridized with the mouse 15K complementary DNA （cDNA） array. The result was analyzed by ArrayVision version 5 and GeneSpring version 5 softwares. After normalization, the ratios of gene expression of parasite treated to untreated NIH-3T3 cells of 2-and more-fold upregulated was defined as the differentially expressed genes. The expression levels of the signal transduction genes were validated by semiquantitative SYBR-based real-time RT-PCR. RESULTS： Among a total of 15 000 genes/ESTs, 239 genes with established cell proliferation-related function were 2 fold-and more-up-regulated by O. viverrini ES product compared to those in cells without exposure to the parasitic product. These genes were classified into groups including energy and metabolism, signal transduction, protein synthesis and translation, matrix and structural protein, transcription control, cell cycle and DNA replication. Moreover, the expressions of serinethreonine kinase receptor, receptor tyrosine kinase and collagen production-related genes were up-regulated by O.viverrini ES product, The expression level of signal transduction genes, pkC, pdgfra, jak 1, eps 8, tgfβ 1/4,strap and h ras measured by real-time RT-PCR confirmed their expression levels to those obtained from cDNA array. However, only the up-regulated expression of pkC,eps 8 and tgfβ3 1/4 which are the downstream signaling molecules of either epidermal growth factor （EGF） or transforming growth factor-β （TGF-β） showed statistical significance （P 〈 0.05）.CONCLUSION： O. viverrini ES product stimulates the significant changes of gene expression in several functional categories and these mainly include transcripts related to cell proliferation. The TGF-β and EGF signal transduction pathways are indicated as the possible pathways of O. viverrini-driven cell proliferation.

EFFECTS OF MEDICINAL MOXIBUSTION ON SERUM GASTRIN,URINARY D-XYLOSE EXCRETORY RATE AND T CELL SUBGROUPS IN SPLEEN DEFICIENCY PATIENTS

The medicinal moxa stick made by moxa wool combining with Chinese medicines of supporting the body resistance and strengthening the spleen was used to treat 74 cases of spleen deficiency, the other 41 patients as control were treated by pure moxa stick. After treatment, the serum gastrin, urinary D - xylose excretory rate and T cell subgroups in patients with spleen deficiency have been greatly improved as compared with those before treatment in two groups (P< 0.01 or 0.05). Comparing the changes of the indexes between treatment group and control group, there were significant(P<0.05), or markedly significant(P<0.01) differences. It indicated that the medicinal moxibustion could improve the secreting and absorbing functions of digestive tract and the cellular immune function in patients with spleen deficiency.

Prospective study of SEVA TB peroxidase assay for cocktail antigen and antibody in the diagnosis of Tuberculosis in suspected patients attending a tertiary care hospital located in rural area

Objective:To evaluate inhouse developed SEVA TB peroxidase enzyme immunoassay using cocktail of mycobacterial excretory-secretory antigens(ES-31,ES-43 & EST-6) for antibody detection and their affinity purified antibodies for antigen detection in tuberculosis suspected patients.Methods:Inhouse developed SEVA TB peroxidase enzyme immunoassay was evaluated prospectively in 73 suspected pulmonary and 46 extra-pulmonary tuberculosis patients during November 2008~March 2009 in a tertiary hospital located in rural area.Results:Assay on prospective analysis showed 100% correlation of pulmonary tuberculosis(PTB) and extrapulmonary tuberculosis(EPTB) acid fast bacilli positivity and antitubercular treatment in 11 cases.Thirty nine PTB and 12 EPTB cases showed negative for EUSA test and were also not given antitubercular therapy.However 30 PTB and 27 EPTB cases showing ELISA positivity were neither acid fast bacilli positive nor antitubercular therapy treated.These cases may possibly have dormant infection and need further diagnosis.In EPTB cases ELISA was observed to be more useful than AFB smear test.Conclusions:This inhouse developed user-friendly peroxidase ELISA can be used as an adjunct test of smear microscopy or culture techniques for routine screening of patients suspected of PTB or EPTB.

Effects of inductive methods on dunging behavior of weaning pigs in slatted floor pens

The excreta of pig is generally utilized to induce pigs to excrete in particular area,which reduces the subsequent work required to clean pens.This paper discussed a new induction device design based on the biological characteristics of pigs.Using different induction materials in the devices,the frequency and location of the excretory behavior of pigs through five treatments and a control group were compared.According to the results,different induction methods had significant(p<0.05)effects on frequency and duration of excretory behavior.Compared to the conventional induction method,the induction devices were significantly(p<0.05)more effective in training pigs to excrete in the assigned area,the most effective material used in the induction devices was feces.If the inductive feces had been preserved in the air for a longer time,the effect of inducing excretory behavior would have been more obvious.Empty devices did not work to induce pigs to excrete in the induction area.