China ; epidemiology ; Humans ; Incidence ; Occupational Diseases ; epidemiology

Objective To investigate the relationship of the expression of DNA methyltransferase,demethylase(mbd2) and tumor-associated genes in human gastric cancer. Methods Tissue samples of cancerous,para-cancerous and normal gastric mucosa were obtained surgically from 28 primary gastric cancer patients. The transcription level of DNA methyltransferase 1 (DNMT1),mbd2,methyl-CpG binding protein (MeCP2),p16 INK4A and c-myc were determined by using real-time RT-PCR and RT-PCR. The relationship between the expression of DNA methylation-associated genes and tumor-associated genes was analyzed. Results The mRNA level of DNMT1 was higher and the mRNA level of mbd2 gene was lower in cancerous tissue than that in normal tissue. The expression of c-myc instead of p16 INK4A and MeCP2 was increased in cancer tissues. The mRNA level of c-myc related negatively to mbd2 when gastric cancer developed. However,there was no any close relation between the transcription level of all above genes and tumor biological behavior in human gastric cancer. Conclusion This study indicates that MeCP2 but not DNMT1 may contribute to the regulation of tumor-associated genes expression in human gastric cancer.

objective To assess the effect of aldosterone on the production of matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9)and collagen Ⅳ in culture supematants of podocytes and the possible molecular mechanisms involved in the influence of aldosterone on the synthesis and degradation of extracellular matrix produced by podocytes. Methods Podecytes were treated with aldosterone at the concentration of 10-11, 10-9, 10-7 mol/L respectively. Cultured podocytes were examined at 24, 48 and 72 hours respectively. Spironolactone, a receptor antagonist of aldosterone, was added to observe the blocking effect on aldosterone. An inhibitor of TGF-β1 receptor was used to determine whether the effect of aldosterone on podocytes were mediated through TGF-β1 system. The enzymatic activities of MMP-2 and MMP-9 were assayed by gehtin zymography. Collagen Ⅳ 0.5 chain and TGF-β1 proteins released into culture supematants were assessed by Western blot and ELISA analysis. The adhesion rate of podocytes was monitored by flow cytometry. Results Aldosterone increased the activities of MMP-2 and MMP-9 in a dose- and time-dependent manner (P<0.05). Aldosterone decreased the level of collagen Ⅳ or5 chain protein in culture supernatants (P<0.05). Meanwhile, the expression of TGF-β1 was also increased (P<0.05). Spironolactone completely abolished the above-mentioned changes(P< 0.05). Blockage of TGF-β1 signaling with SB431542 prevented the aldosterone-induced upregulation of MMP-2 and MMP-9 as well as the downregulation of the collagen Ⅳ α5 chain protein and the adhesion rate of podocytes (P<0.05). Conclusions Aldosterone increases the activities of MMP-2 and MMP-9 but decreases the expression of collagen Ⅳ α5 chain and the adhension rate of podocytes possibly via TGF-β1 signaling pathway. Such alterations may contribute to glomerular podocyte injury associated with the GBM abnormality caused by the imbalance between matrix synthesis and degradation.

Objective To investigate the clinical therapeutic effect of filiform fire needling on tennis elbow.Method Eighty patients with tennis elbow were randomly allocated to a filiform fire needling group of 30 cases, a block group of 20 cases and an electroacupuncture group of 30 cases. The treatment group received. Pain severity was scored using the VAS after one and two weeks and one month of treatment. The therapeutic effects were evaluated and compared after one month of treatment.Result The total efficacy rate was 93.3% in the filiform fire needling group, 60.0% in the block group and 50.0% in the electroacupuncture group; there was a statistically significant difference among the three groups (P<0.05). There was a statistically significant difference between the filiform fire needling and block groups (P<0.05) and between the filiform fire needling and electroacupuncture groups (P<0.05) but no significant difference between the block and electroacupuncture groups (P>0.05). The VAS score decreased significantly in the block group (P<0.05) but did not in the filiform fire needling and electroacupuncture groups (P>0.05) after one week of treatment compared with before. The VAS score decreased significantly in the filiform fire needling and block groups (P<0.05) but did not in the electroacupuncture group (P>0.05) after two weeks of treatment compared with before. There was no statistically significant posttreatment difference in the VAS score between the filiform fire needling and block groups (P>0.05) but there was such a difference between the filiform fire needling or block group and the electroacupuncture group (P<0.05). There was a statistically significant difference in the VAS score in the filiform fire needling and block groups (P<0.01) and also in the electroacupuncture group (P<0.05) between before and after one month of treatment. There was a statistically significant difference in the VAS score after one month of treatment between the filiform fire needling group and the block or electroacupuncture group (P<0.05) and between the block and electroacupuncture groups (P<0.05). The results of the study showed that filiform fire needling, local drug blockage and electroacupuncture all had clinically a better therapeutic effect on tennis elbow and a better improving effect on the symptoms in the patients. Filiform fire needling produced a marked therapeutic effect. Its effect was superior to those of block therapy and electroacupuncture.Conclusion Filiform fire needling is a definitely effective way to treat tennis elbow.

To investigate the protective effects of eplerenone on adriamycin-induced renal injury and the possible mechanisms involved, 36 male Sprague-Dawley rats were randomly divided into control group, adriamycin nephropathy (AN) group and eplerenone-treated group (100 mg·kg(-1)·d(-1) eplerenone). Blood pressure, 24-h urinary protein, serum potassium, sodium and creatinine were measured 28 days after adriamycin injection (a single tail intravenous injection of 6.5 mg/kg adriamycin). The morphological changes of renal tissues were observed by light and electron microscopy. Immunohistochemistry and Western blotting were performed to examine the expression of TGF-β(1) and desmin in renal cortex. The results showed that 28 days after adriamycin injection, there were no significant changes in the level of serum potassium, sodium, creatinine concentrations and blood pressure values in the rats of the three groups. Meanwhile, the 24-h proteinuria excretion in the AN group was significantly higher than that in the control group (P<0.01), but that in the eplerenone-treated group was substantially reduced when compared with that in the AN group (P<0.05). Mild mesangial cell proliferation and matrix expansion, diffuse deformation and confluence of foot processes in podocytes were found in the AN group. By contrast, rats in the eplerenone-treated group exhibited obvious attenuation of these morphological lesions. The protein expression of TGF-β(1) and desmin in the AN group was markedly up-regulated in contrast to that in the control group (P<0.01), whereas that in the eplerenone-treated group was much lower than in the AN group (P<0.05). It was concluded that eplerenone may ameliorate the proteinuria and the development of pathological alteration in adriamycin-induced nephropathy presumably via the inhibition of cytokine release, and restore the morphology of podocytes independent of its blood pressure-lowing effects.

Objective To analyze the effect of eukaryotic expression vector containing sense and antisense DNA methyltransferase (DNMT1) gene on the transcript level of tumor associated genes in human colon cancer cell line. Methods Recombinant plasmid, including sense DNMT1 (HMT) and antisense DNMT1 (THM) gene, were constructed and transfected into SW1116 cell by using the lipofectamine. Then G418 filtration was performed. The expression of DNMT1 protein was examined by Western blotting. The transcription level of hMLH1, hMSH2, c myc and p16 INK4A genes were detected by RT PCR. Results The sense and antisense eukaryotic expression vectors were successfully constructed and then the constructed recombinant plasmids were transfected into SW1116 cell. The protein levels of DNMT1 have been up regulated and down regulated in SW1116 cells transfected with HMT and THM plasmids, respectively. The mRNA level of hMLH1, hMSH2, c myc gene were down regulated in the sense DNMT1 transfected cell. The mRNA level of hMSH2 was up regulated in the antisense DNMT1 transfected cell. However, the transcription level of p16 INK4A gene could not be associated with DNMT1 in SW1116 cell.Conclusion DNMT1 can regulate the expression of the tumor associated genes in human colon cancer cell line SW1116.

OBJECTIVE: To investigate the effects of Erxian Decoction (EXD), a traditional Chinese compound herbal medicine and its separate prescriptions such as Wenshen Yijing Recipe (WSYJR, a recipe for warming kidney and replenishing essence), Ziyin Xiehuo Recipe (ZYXHR, a recipe for nourishing yin and dispersing fire) and Tiaoli Chongren Recipe (TLCRR, a recipe for regulating thoroughfare and conception vessels) and some extracts from EXD on the levels of LH and FSH in the primary cultured anterior pituitary cells from female rats. METHODS: EXD, its separate prescriptions and traditional Chinese herbal extracts from EXD were added directly to the incubators. The levels of LH and FSH were tested by radioimmunoassay. RESULTS: The levels of LH and FSH in the supernatant of anterior pituitary cells treated by EXD or its separate prescriptions including ZYXHR and TLCRR were increased significantly as compared with those in the blank control. There was a tendency for stimulating the secretions of LH and FSH in the WSYJR-treated group too. Dimethyl sulfoxide as the solvent of icariin and curculigoside could interfere with the results. CONCLUSION: EXD and its separate prescriptions such as ZYXHR and TLCRR can increase the levels of LH and FSH significantly.

Objective To study the effect of overexpression of TRPC6 on Ang Ⅱ-induced apoptosis of mouse podocytes in vitro and to explore the possible mechanisms. Methods Mouse TRPC6 cDNA eukaryotie expression vector pEGFP-NI-mTRPC6 was transfected to conditionally immortalized routine podocyte cell line by liposome. The fluorescent microscopy was used to examine the expression of EGFP after 24 hours. The change of TRPC6 protein expression was observed by Western-blot. Podocytes were treated by different concentrations of Ang Ⅱ. The podocyte intracellular calcium concentration was measured with laser-scanning con_focal microscope. The expression of Bax and Bcl-2 mRNA was assessed by RT-PCR and the expression of Bax and Bcl-2 protein was measured by Western-blot. The apoptotic ratio of podocytes was monitored by flow cytometry and Hoechst staining. Results About 35% of the cells expressed EGFP. An up-regulation of protein expression of TRPC6 was detected in podocytes when transfected with pEGFP-N1-mTRPC6 (P<0.01). The overexpression of TRPC6 promoted the Ang Ⅱ-induced influx of extracellular calcium and elevated the expression of Bax but decreased the expression of Bcl-2 (P<0.01, P<0.05). The apoptotic ratio of podocyte was (2.50±0.72)% when treated by low-dose Ang Ⅱ (10-10 mol/L), and it was increased to (4.33±0.45)% when transfected with pEGFP-N1-mTRPC6 (P <0.05 ). Transfection with pEGFP-NI-mTRPC6 increased apoptosis rate from (15.46± 1.40)% to (18.33±0.87)%(P<0.01) by high-dose Ang Ⅱ (10-6 mol/L). Conclusion TRPC6 plays an important role in the Ang Ⅱ-induced apoptosis of podocytes by promoting the influx of extraeellular calcium, which leads to the apoptosis cascade initiation.

Objective To study the mutations of outer-membrane porin gerte (oprD) in imipenem-resistant Pseudomonas aeruginosa.Methods The PCR was applied to detect the oprD gene from the 34 clinical imipenem-resistant Pseudomonas aeruginosa.DNA sequence was proceeded to analysis the nuclentide sequence of the oprD gene and the deduced amino acid sequence.To analysis the mutation and the function of the oprD domain,those mutations were compaired with the standard Pseudomonas aeruginosa ATCC27853 and 2 clinical imipenem-susceptibility isolates.Results oprD gene mutation was wide and diverse.The rate of the mutation was 92.3% (12/13),mutations were concluded dot mutation,deletion mutation and insert mutation,those result in the amino sequence change and frame shift in L2 and L3 loops of outer membrane protein D,hampering the combine of oprD and imipenem.Some new mutations were found.They were 1 079,1 114,1 196,1 206,1 288,1 300,1 301 bases and 115,127,154,158,185,189 aminos.All above mutations were not deteced in ATCC 27853 and 2 clinical imipenem-susoeptibility isolates.Conclusions The wide and diverse mutations in oprD gene result in amino acid change and/or frame shift L2 and L3 loops,hampering the binding of IMP and oprD.Those may result in resistance to imipenem in Pseudomonas aeruginosa.

ObjectiveTo observe gene transcription characteristic of AC-cAMP signal pathway in thyroid of H22 tumor-bearing mice with different syndromes.MethodsThe quantitative four diagnosis and syndrome differentiation methods and Affymetrix Gene Chip Mouse Exon 1.0 ST Array were used,thyroid gene expression of normal mice,qi-deficiency syndromes (QDS) and poisonous pathogenic factors syndrome (PPFS) in early stage of H22 tumor-bearing mice were detected.Genes of AC-cAMP signal pathway in gene chips,which related with thyroid hormone synthesis and secretion were selected,and then analyzed their expressive characteristic.Results ①In key genes,TG and Pax8 were down-regulated in early stage,TSHr down-regulated in PPFS while contrary in QDS.②the transcription level of most genes in QDS were slightly higher than in PPFS,Nis,Tpo,AC,Ttf1,Titf2 and Prkaca were included.()Slight changes showed in other genes in this pathway.ConclusionIn AC-cAMP pathway of H22 tumor-bearing mice with different symptoms,some key genes showed similar characteristics including TSHr,AC,Pax8,Ttf1,Tiff2,TG,Nis and Tpo.This suggested that the thyroid is inhibited in mice with PPFS.