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.

AIM: To explore the feasibility of direct separat and selective enlargement of the bone marrow-derived liver stem cells (BDLSC) from bone marrow cells with a culture system containing cholestatic serum in vitro . METHODS: Bone marrow cells of rats were cultured with selective media containing 2%, 5%, 7% and 10% cholestatic rat serum, respectively. The BDLSC were then induced to proliferate with the addition of hepatocyte growth factor (HGF) on the firth day. BDLSC were characterized using immunocytochemistry and RT-PCR for lineage markers, glycogen staining and urea synthetic assay for functions 2 weeks later. RESULTS: Bone marrow cells were unble to form colony in the presence of 2% cholestatic serum and apopotosis appeared gradually in 7% or 10% cholestatic serum. The BDLSC survived in the medium containing 5% cholestatic serum while the other types of cells did not. The survival cells proliferated with a high speed during the second week and then formed hepatocyte-like colony-forming units (H-CFU). Cells in the H-CFU expressed the characteristic proteins of fetal hepatocytes. Furthermore, they had glycogen storage and urea synthesis functions, two of the critical features of hepatocytes. CONCLUSION: The selective micro-environment effectively selected BDLSC from the bone marrow cell, and will be a new way to provide an abundant source of donor hepatocytes for clinical cell therapy.

Objective To study the rhubarb phenol and P38 inhibitor suppression on light aging of human skin HaCaT induced by UVB. Methods The potential of cell proliferation of the different concentrations of rhubarb phenol and P38 inhibitor (SB203580) on human skin HaCaT was detected by MTT method. The cells was divided into the control group, the model group, the rhubarb phenol group, the SB203580 group by random number table method after the 24 h incubation. The 10-6 mol/L rhubarb phenol and 10-7 mol/L SB203580 were added to the rhubarb phenol group and SB203580 group for 24h, The competence of cultured cell proliferation which was irradiated with UVB of intensity of 0.61mW/cm2, mutiply time of 7 min and distance of 10 cm for 24 h except the control group; Western Blot method detected rhubarb phenol and P38 inhibitor of the influence of P38, P-P38, TNF-α, IL-6 protein. Results Compared with control group, the cell proliferation in UVB group significantly reduced (P<0.01); Compared with UVB group, the expression of the P-P38 (0.419 ± 0.029, 0.398 ± 0.015 vs. 0.497 ± 0.051), TNF-α (0.435 ± 0.025, 0.411 ± 0.021 vs. 0.509 ± 0.040) and IL-6 (0.457 ± 0.027, 0.432 ± 0.018 vs. 0.478 ± 0.036) in rhubarb phenol and P38 inhibitor group significantly reduced (P<0.01). Conclusions The rhubarb phenol and P38 inhibitor could significantly suppress HaCaT cells light aging, and its mechanism may be related with inhibiting P38 signaling pathways, and inhibiting the secretion of inflammatory cytokines.

To investigate the effects of albumin on the production of matrix metalloproteinases-2 (MMP-2) and matrix metalloproteinases-9 (MMP-9) in podocytes. Podocytes were treated with bovine serum albumin (BSA) at the concentration of 0.1, 0.5, 1, 2 g/L, respectively. Conditioned media were harvested 12, 24, 48 and 72 h after the treatment. The expression of MMP-2 and MMP-9 was assayed by gelatin zymography, RT-PCR and Western blotting analysis. Our results showed that in comparison with the control group, BSA increased the expression of MMP-2 and MMP-9 mRNA and protein in a dose- and time-dependent manner (P<0.05). Meanwhile, the enzymatic activities of MMP-2 and MMP-9 in the culture supernatants of podocytes were also increased (P<0.05). It is concluded that albumin up-regulated the expression of MMP-2 and MMP-9 at gene and protein levels in a time- and dose-dependent manner.

This study examined the effect of sulodexide on podocyte injury in rats with adriamycin nephropathy (AN). A total of 36 healthy male SD rats were randomly assigned to three groups: control group, AN group and sulodexide treatment group. Rat models of AN were established by a single tail intravenous injection of adriamycin (6.5 mg/kg) in both AN group and sulodexide treatment group. Sulodexide (10 mg/kg) was administered the rats in the treatment group once daily by garage from the first day of model establishment until the 14th day or the 28th day. Samples of 24-h urine and renal cortex tissues were harvested at day 14, 28 after the model establishment. Excretion of 24-h urinary protein was measured by Coomassie brilliant blue method. The pathological changes in renal tissues were observed by light microscopy and electron microscopy respectively. Heparanase mRNA was detected by RT-PCR. Expressions of desmin, CD2AP and heparanase were determined by immunohistological staining. The results showed that the expressions of heparanase mRNA and protein were increased in the glomeruli of AN rats at day 14 and 28 after the model establishment, which was accompanied by the increased expression of desmin and CD2AP. The mRNA and protein expression of heparanase was decreased in the sulodexide-treated rats as compared with AN rats at day 14 and 28. And, the protein expression of desmin and CD2AP was reduced as with heparanase in the sulodexide- treated rats. Proteinuria and podocyte foot process effacement were alleviated in the AN rats after sulodexide treatment. There was a positive correlation between the expression of heparanase and the expression of desmin and CD2AP (as well as 24-h urinary protein excretion). It was concluded that increased heparanase is involved in podocyte injury. Sulodexide can maintain and restore podocyte morphology by inhibiting the expression of heparanase in AN.

Eukaryotic expression vectors carrying the small hairpin RNA (shRNA) for TRPC6 mRNA were constructed, and the effects of knocking-down TRPC6 on puromycin aminonucleoside (PAN)-induced apoptosis of mouse podocytes were observed. Two eukaryotic expression vectors containing small hairpin structure targeting TRPC6 named pGCsi-TRPC6A and pGCsi-TRPC6B were designed and synthesized. The plasmids were transfected into conditionally immortalized murine podocyte cell line by liposome. The changes in the TRPC6 mRNA and protein expression were observed by RT-PCR and Western blot after 48 h. Cultured podocytes were divided into four groups: control group, PAN treatment group, PAN treatment+shRNA transfection group, and PAN treatment+ negative control group. The expression of Bax and Bcl-2 mRNA and proteins was detected by RT-PCR and Western-blot respectively. The apoptotic rate of podocytes was measured by flow cytometry. The results showed that the expression of TRPC6 mRNA and protein was decreased in the podocytes when transfected with pGCsi-TRPC6A, and pGCsi-TRPC6B. The expression of Bax was increased, and that of Bcl-2 was decreased at protein and mRNA levels in the podocytes after treated with PAN for 48 h. These changes was attenuated by knocking-down TRPC6. Knocking-down TRPC6 could effectively decrease the PAN-induced apoptosis of podocytes. It was concluded that TRPC6 may play an important role in the PAN-induced apoptosis of podocytes. Knocking-down TRPC6 gene could effectively prevent the podocytes from apoptosis induced by PAN.

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.