An epidemiologic study on functional constipation among adult communities in Shanghai / 中华流行病学杂志

Objective To determine the prevalence and risk factors of functional constipation (FC) by using Rome Ⅲ criteria in the local adult communities.Methods A stratified randomized and community-based study by multi-stage cluster sampling was employed.A household survey was conducted from April to May 2010.All of the participants were interviewed face-to-face by filling out the self-administered questionnaires which based on Rome Ⅲ criteria for the diagnosis of FC.Self-rating anxiety scale (SAS),self-rating depression scale (SDS) and Athens insomnia scale (AIS) were carried out to evaluate the psychological characteristics and qualities of sleep.Results A total of 7648 subjects fulfilled the questionnaires,with the response rate as 90.0％.211 patients met the Rome Ⅲ criteria,including 90 males and 121 females.The adjusted prevalence rates of FC were 2.5％ in males,3.3％ in females and with an overall rate as 2.9％.The ratio of men to women was 1∶1.32,with significant difference between males and females (P=0.043).The most common group was in the 18-29 year-olds (x2=37.359,P=0.000).FC patients were more likely to be detected in the group with normal BMI (x2=16.087,P=0.002),having received high education (x2=27.604,P=0.000),being intelectuals ( x2=6.922,P=0.031 ) and divorced ( x2=22.000,P=0.000) than in other groups. Multivariate analysis showed that excessive intake of high-fat food was significantly associated with the presence of FC (odds ratio as 1.253,P=0.000),whereas foods with high-fiber (odds ratio as 0.854,P=0.029) might serve as protective factors.Significant differences between FC groups and control groups were found in the incidence of anxiety (with odds ratio as 2.583,P=0.000) and insomnia (odds ratio as 2.443,P=0.000).Conclusion The prevalence of FC in adult communities in Shanghai Songjiang district was not higher than that in other parts of the communities.Excessive intake of high-fat food,anxiety and insomnia might be risk factors for FC and foods with high-fiber contents might serve as protective factors.

Effect of silencing connective tissue growth factor on the liver fibrosis in rats / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the anti-fibrogenesis property of intraportal vein small interfering RNA (siRNA) injection targeting connective tissue growth factor (CTGF) in a rat model of liver fibrosis induced by carbon tetrachloride (CCl4) and its effect on hepatic stellate cell (HSC) activation.</p><p><b>METHODS</b>24 male rats were randomly divided into four group. rats received CCl4 by subcutaneous injections every three days for 6 consecutive weeks, and meantime they also obtained either siRNA targeting CTGF (as CTGF siRNA group), saline (as model group) or a control siRNA (as control siRNA group) by intraportal vein injection to rats liver at the same approach. Other rats received saline intraportal vein injection for 6 weeks (as normal control group). The expressions of CTGF and a-SMA protein were detected by Western blot. Hepatic histology was evaluated by HE staining and Sirius red staining. The collagen staining areas were measured quantitatively using a computer-aided manipulator with slight modifications. The number of active HSC were evaluated by immunohistochemistry.</p><p><b>RESULTS</b>Six weeks after CCl4 injection, prominent upregulations were observed in the expressions of CTGF and a-SMA protein in saline or control siRNA-treated rats livers. In rats with CTGF siRNA treatment, the protein expressions of CTGF and a-SMA in liver decreased by 95%+/-2% and 86%+/-11% (F=21.234 and 12.473, P<0.01) respectively, the number of active HSC in liver decreased by 76%+/-9% (F=9.179, P<0.01) as compared to the model group. The attenuation of liver fibrosis was also observed in rats with CTGF siRNA treatment.</p><p><b>CONCLUSION</b>Intraportal vein siRNA injection targeting CTGF could significantly inhibit CTGF gene expression in rats, thereby attenuate liver fibrosis by decreasing the number of active HSCs.</p>

Effect of interleukin 10 gene-modified bone marrow-derived liver stem cells transplantation on hepatic inflammatory response and liver regeneration in hepatic fibrosis rats / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To explore effect of interleukin 10 (IL-10) gene-modified bone marrow-derived liver stem cells (BDLSCs) transplantation on hepatic inflammatory response and liver regeneration in rats with liver fibrosis.</p><p><b>METHODS</b>50 female Wistar rats were randomly divided into 4 groups: (1) control group: 10 rats were subcutaneously injected with olive oil for 8 weeks; (2) fibrosis groups: 16 rats were subcutaneously injected with carbon tetrachloride (CCl4) for 8 weeks to induce liver fibrosis; (3) BDLSC group: 12 rats were subcutaneously injected with CCl4 for 8 weeks, and were transplanted with 2 x 10(5) BDLSC at week 4; (4) BDLSC/IL-10 group: 12 rats were subcutaneously injected with CCl4 for 8 weeks, and were transplanted with 2 x 10(5) IL-10 gene-modified BDLSC at week 4. IL-10 and tumor necrosis factor alpha (TNFa) in liver tissues were detected by ELISA. HE stained liver tissues were observed under light microscope. The expression of hepatocyte growth factor (HGF) was quantified by real-time RT-PCR, and the expression of proliferating cell nuclear antigen (PCNA) was determined by immunohistochemistry.</p><p><b>RESULTS</b>The ratio of IL-10/TNFa in fibrosis group (0.05+/-0.01) was lower than that in control group (0.26+/-0.04) (P < 0.01). Transplantation of untreated BDLSCs did not improve the ratio (P > 0.05), however, transplantation of IL-10 modified BDLSCs improved the ratio significantly (P < 0.01). Severe inflammatory response and fibrosis were observed in fibrosis group. Inflammatory response was alleviated to some extent in the BDLSC group, and the histopathology of BDLSC/IL-10 group was not significantly different from that of the control group. Compared to the control group, the expression of HGF mRNA and PCNA protein was increased in the fibrosis group (P < 0.01). The expression of HGF and PCNA was further increased by BDLSCs or IL-10 modified BDLSCs transplantation. Compared to BDLSCs, IL-10 gene-modified BDLSCs were more potent to induce the expression of HGF and PCNA.</p><p><b>CONCLUSION</b>Transplantation of IL-10 gene-modified BDLSCs can alleviate hepatic inflammatory response and promote liver regeneration in hepatic fibrosis rats.</p>

Effect of silencing connective tissue growth factor on rat liver fibrosis and the accumulation of extracellular matrix / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the anti-fibrogenesis property of intraportal vein injection of small interfering RNA targeting connective tissue growth factor (CTGF) in a rat model of liver fibrosis and its effect on the accumulation of extracellular matrix (ECM).</p><p><b>METHODS</b>Thirty male rats were randomly divided into five groups. Some rats received CCl4 subcutaneously every three days for 6 consecutive weeks, and in the meantime they also received either siRNA targeting CTGF (preventive group), saline (model group) or siRNA (siRNA control group) by intraportal vein injections. Other rats received CCl4 by subcutaneous injection for 2 weeks, followed by CCl4 and CTGF siRNA intraportal vein injection for 4 more weeks (as treatment group). The expressions of CTGF and type I and III collagen genes were detected by means of reverse transcription-polymerase chain reaction (RT-PCR) and/or Western blot respectively. Hepatic histology was evaluated by HE and Sirius red stained sections. The collagen staining areas were measured quantitatively using a computer-aided manipulator with slight modifications. Serum procollagen type III and hyaluronic acid were determined by radioimmunoassay.</p><p><b>RESULTS</b>Six weeks after CCl4 injection, prominent upregulation of gene expressions of CTGF, type I and III collagen, and laminin in saline or siRNA-treated rat livers were observed. The expressions of CTGF at mRNA and protein level and type I and III collagen at mRNA level were markedly reduced in rats with CTGF siRNA treated for four or six weeks. Expressions of CTGF at mRNA and protein levels decreased by 76%+/-8%, 80%+/-3% (F = 68.630) and 95%+/-2%, 93%+/-3% (F = 21.234, P < 0.01); type I and III collagen and laminin at mRNA levels decreased by 74%+/-8%, 78%+/-8%, 31%+/-7% and 57%+/-6%, 59%+/-10%, 43%+/-9% (F = 24.219, 16.315, 9.716, P < 0.01) compared with rats in the model group at 72 h. The CTGF siRNA treatment markedly reduced serum levels of procollagen type III and hyaluronic acid and the degrees of liver fibrosis.</p><p><b>CONCLUSION</b>Intraportal vein siRNA injection targeting CTGF could significantly inhibit CTGF gene expression in rats, thereby attenuating liver fibrosis by reducing ECM accumulation.</p>

Effects of silencing connective tissue growth factor on rat transforming growth factor beta/Smads signal / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the effects of small interfering RNA targeting connective tissue growth factor (CTGF) on rat transforming growth factor beta (TGF beta)/Smads signal pathway.</p><p><b>METHODS</b>Chemically synthetic siRNA targeting CTGF was transfected into HSC T6 and then they were injected into rat livers through their intraportal veins. At the same time these rats also received CCl4 subcutaneously every three days for 6 consecutive weeks. Untreated HSC T6 or/and rats with random siRNA treatment served as controls. Total RNA or/and protein in HSC T6 and rat hepatic tissues were extracted. The expressions of CTGF and TGF beta 1, Smad2, 3 and 7 genes were detected by reverse transcription-polymerase chain reaction (RT-PCR) and/or Western blot.</p><p><b>RESULTS</b>CTGF siRNA significantly reduced the expression of CTGF protein in HSC T6. At 48 h after CTGF siRNA treatment, the down-regulation of CTGF protein was the most significant, up to 94%+/-4% (t=46.196, P less than 0.01), but the expressions of TGF beta 1, Smad2, 3 and 7 mRNA showed no differences in HSC T6 compared with the blank controls. Six weeks after CCl4 injections, prominent up-regulations were observed in the gene expressions of CTGF and TGF beta 1 in saline control or siRNA-treated rat livers. Administering CTGF siRNA for six weeks markedly attenuated the induction of CTGF and TGF beta 1 genes; the expressions of CTGF and TGF beta 1 protein decreased by 95%+/-2% (F=21.234, P less than 0.01) and 74%+/-8% (F=13.464, P less than 0.05), respectively, whereas Smad2, 7 protein expressions were not affected.</p><p><b>CONCLUSION</b>Silencing the CTGF gene can suppress the TGF beta /Smads signal pathway in rat livers.</p>

Risk factors of irritable bowel syndrome in adolescents in China / 中华儿科杂志

<p><b>OBJECTIVE</b>To explore the risk factors for irritable bowel syndrome (IBS) among school adolescents in China.</p><p><b>METHOD</b>A stratified, randomized study by cluster sampling was conducted, which recruited 51,956 students from high and primary schools in Chinese cities. All students were requested to fill in a questionnaire.</p><p><b>RESULT</b>(1) Factors including class (odds ratio 1.12), excessive intake of pepper (odds ratio 1.17), fried (odds ratio 1.08) and starch-based foods (odds ratio 1.06), gastrointestinal tract infection (odds ratio 2.66), abuse of analgesic (odds ratio 1.49), inheritance (odds ratio 1.83), fatigue (odds ratio 1.32) and repression (odds ratio 1.45) were significantly associated with the presence of IBS (P < 0.05). High protein food (odds ratio 0.90) was a protective factor.</p><p><b>CONCLUSION</b>Different food intake, gastrointestinal tract infection, abuse of analgesic, inheritance and psychological factors might be related to development of IBS in the students of the cities involved in this study.</p>

In vitro study of transfection of mesenchymal stem cells with adenoviral vector overexpressing human uPA / 上海第二医科大学学报

Objective To identify the isolated rat bone marrow derived mesenchymal stem cells(MSCs),and evaluate the efficiency of adenoviral vector expressing human urokinase type plasminogen activator(uPA) in transfection of rat MSCs and its effect on proliferation of MSCs. Methods MSCs were isolated and purified by pasted wall purification,and were identified by immunicytochemistry.The transfection efficiency of uPA was detected by fluorescent microscopy,the expression of uPA in MSCs was detected by Western blotting,and the proliferation of MSCs was evaluated by MTT. Results The harvested MSCs exhibited the typical appearance of MSCs,and it was revealed by immunohistochemistry that the expression of MSCs markers CD29 and CD90 was positive,while that of CD34 and CD45 was negative.A tendency of increase in expression of green fluorescent protein(GFP) was observed with increase of multiplicity of infection(MOI).After transfection with AduPA for 72 h,the transfection efficiency reached(94.0?1.5)% at MOI of 80,and positive GFP cells could still be observed even after 7 d.The transfected uPA had no effect on the proliferation of MSCs. Conclusion MSCs are favourable genetic vectors to express uPA,and can be used for treatment of liver fibrosis.

Effects of AcSDKP on proliferation and secretion of extracellular matrix in rat active hepatic stellate cells / 上海第二医科大学学报

Objective To study the effects of N-acetyl-seryl-aspartyl-lysyl-proline(AcSDKP) on proliferation and synthesis and secretion of extracellular matrix(ECM) in rat active hepatic stellate cells(HSCs). Methods Primary cultures of HSCs when actived were subjected to AcSDKP(0.01, 0.1, 1, 10 and 100 nmol/L, respectively) treatment, and control group was established. The expression of collagen Ⅰ(ColⅠ) and collagen Ⅲ(Col Ⅲ) mRNA were analyzed with RT-PCR. The proliferation of HSCs was detected by MTT. Hyaluronic acid (HA) and laminin(LN) in the supernatant were determined by enzyme-linked immunosorbent assay. Results Compared with control group, the proliferation of active HSCs was significantly inhibited by 1 and 10 nmol/L AcSDKP. 1, 10 nmol/L AcSDKP and 0.1 to 100 nmol/L AcSDKP significantly inhibited the expression of HSCs ColⅠ mRNA and Col Ⅲ mRNA, respectively. Expression of HA and LN in the supernatant were significantly inhibited by 0.1, 1, 10 nmol/L and 0.1, 1 nmol/L AcSDKP, respectively. ConclusionAcSDKP can inhibit synthesis and secretion of ECM in active HSCs in a dose-dependent manner, with a maximum inhibition effect at 1 nmol/L AcSDKP. The mechanism may involve the inhibition of the proliferation of HSCs,which leads to the decrease of HSCs that synthesize and secrete ECM.

Isolation and characterization of ?_2m~-/Thy-1~+ bone marrow-derived liver stem cells from cholestatic rats in vitro / 上海第二医科大学学报

Objective To explore the in vitro isolation of ?2m-/Thy-1+ bone marrow-derived liver stem cells(BDLSCs) which bear double features of stem and liver cells from bone marrow stem cells(BMSCs)as so to provide suitable donor cells for the treatment of liver diseases by cellular transplant. Methods ?2m-/Thy-1+ BDLSCs were isolated by magnetic bead cell sorting(MACS) method from cholestatic rats in vitro,and cell purity was detected using flow cytometry.Liver associated phenotype markers were characterized by RT-PCR and immunofluorescence staining. Results BDLSCs isolated by MACS were purified and viable,and possessed hepatocyte-like features at gene and protein levels. Conclusion ?2m-/Thy-1+ BDLSCs are special subsets of BMSCs which may have promising potentials in the stem cell-based treatment of liver diseases.

COX-2 expression in the H. pylori infected gastric mucosal epithelia and its significance / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To study COX-2 expression in H. pylori infected gastric mucosal epithelia and its significance in the carcinogenesis of the stomach.</p><p><b>METHODS</b>Rapid urease test and histological examination with basic magnenta staining were used to assess the status of H. pylori infection in the stomach. COX-2 was detected immunohistochemically.</p><p><b>RESULTS</b>COX-2 immunostaining was positive in 1 out of 12 cases with H. pylori-negative gastric mucosa and also in 1 out of 10 cases with H. pylori-positive gastric mucosa without macroscopic alterations, while COX-2 expression was found to be positive in 5 out of 9 cases with H. pylori related superficial gastritis with mucosal erosions. COX-2 expression was detected in 5 out of 10 cases with H. pylori-positive mild atrophic gastritis, 8 out of 10 cases with H. pylori-positive moderate-severe atrophic gastritis and intestinal metaplasia, and 6 out of 8 cases with H. pylori-positive moderate-severe dysplasia. COX-2 expression was positive in 22 out of 32 cases of gastric cancer.</p><p><b>CONCLUSION</b>H. pylori may induce COX-2 expression of gastric mucosal epithelia in chronic superficial gastritis, which is related to the development of mucosal injury. According to gastric mucosal carcinogenesis pattern up-regulation of COX-2 expression is associated with gastric mucosal carcinogenesis, and involved in the early development of premalignant lesions.</p>

Relationship between somatostatin receptors and activation of hepatic stellate cells / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Somafostatin receptors (SSTRs) have been suggested to involve in mediating the effect of somatostatin on hepatic stellate cells (HSCs) in an activation-dependent way. We, therefore, try to investigate the relationship between expression of SSTRs and activation of rat HSCs.</p><p><b>METHODS</b>HSCs were isolated from rats by in situ perfusion and single-step density gradient centrifugation. SSTR1-5 mRNA levels in the differentiated first passage HSCs were detected by means of a reverse transcription polymerase chain reaction. On the other hand, hepatic fibrosis was induced in adult male Sprague-Dawley rats by carbon tetrachloride intoxication, and the expression of SSTR1-5 in normal as well as fibrotic livers was measured by immunohistochemical staining.</p><p><b>RESULTS</b>SSTR mRNA and SSTR could not be found in freshly isolated rat HSCs or normal rat liver. However, SSTR1-3 mRNA appeared as HSCs became wholly activated, and could also be identified on the membrane of activated HSCs in the perisinusoid space, fibrous septa, etc.</p><p><b>CONCLUSION</b>The expression of SSTR1-3 in the rat HSC is closely related to its activation. This may reflect one of the main negative regulation mechanisms in the course of HSC activation.</p>

Effect of small interfering RNA targeting connective tissue growth factor on the synthesis and secretion of extracellular matrix in hepatic stellate cells / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the effect of chemically synthetic small interfering RNA (siRNA) targeting connective tissue growth factor (CTGF) on the synthesis and secretion of extracellular matrix (ECM) in hepatic stellate cells (HSC).</p><p><b>METHODS</b>Chemically synthetic siRNA targeting CTGF was transfected into HSC T6 (an active HSC line in rats) by oligofectamine package, and untreated HSC T6 were used as control. Total RNA and protein of the cells, after their incubation with siRNA for 24, 48 and 72 hours, were extracted, and the supernatants were collected. The expressions of CTGF and type I and III collagen genes were detected by means of reverse transcription-polymerase chain reaction (RT-PCR) and/or Western blot. Contents of hyaluronic acid and type III pro-collagen in the supernatants were determined by radioimmunoassay.</p><p><b>RESULTS</b>The expression of CTGF at mRNA and protein level and type I and III collagen at mRNA levels were markedly down-regulated in siRNA-transfected HSCs. The contents of hyaluronic acid and type III pro-collagen in the supernatants decreased by 46%+/-7%, 52%+/-7%, 53%+/-7% and 29%+/-18%, 29%+/-7%, 27%+/-5%, compared with those of the blank control at 24, 48 and 72 hours.</p><p><b>CONCLUSIONS</b>Chemically synthetic anti-CTGF siRNA can significantly inhibit CTGF gene expression in HSC, and markedly reduce the synthesis and secretion of ECM including type I and III collagen and hyaluronic acid. The siRNA-directed suppression of CTGF gene in HSC was maintained for 72 hours. This suggests that chemically synthetic siRNA may be a potential in preventing and treating liver fibrosis and may have a promising future for development</p>

Therapeutic effects of octreotide on hepatofibrosis-induced with carbon tetrachloride in rats / 中华肝脏病杂志

<p><b>OBJECTIVES</b>To investigate the therapeutic effects and mechanism of octreotide on experimental hepatic fibrosis in rats.</p><p><b>METHODS</b>Hepatofibrotic rats models were established with carbon tetrachloride. All the experimental rats were divided into four groups: normal control group, pre-and post-treatment model group, and octreotide-treated group in which the rats were injected subcutaneously with octreotide at the dose of 50ng/100g, twice daily, for thirty days. Serum levels of hyaluronic acid (HA), laminin (LN) and pro-collagen type III peptide (PCIII) were detected by radioimmunoassay. Hepatic fibrosis scoring grade was assessed through Van-Gieson staining and observed under light microscope. Protein expression levels of alpha-smooth muscle actin (alpha-SMA) and transforming growth factor beta1 (TGFbeta1) were determined with immunohistochemical staining method. Messenger RNA (mRNA) levels of collagen type I and PCIII were detected by reverse transcription polymerase chain reaction.</p><p><b>RESULTS</b>Serum levels of HA (ng/L), LN (microg/L) and PCIII (ng/L) in pre- and post-treatment model groups were higher than those in normal control group (121.8+/-9.5 and 110.3+/-13.4 vs. 33.1+/-3.7, 85.7+/-12.1 and 78.2+/-7.9 vs. 37.1+/-6.3, 35.9+/-3.5 and 33.7+/-2.6 vs. 15.6+/-2.8, respectively, t > or = 9.41, P<0.05), and there was no significant difference between the two model groups. Concentrations of HA (55.8ng/L+/-7.2ng/L), LN (43.1microg/L+/-3.4microg/L) and PCIII (27.8ng/L+/-3.4ng/L) decreased significantly in octreotide-treated group, compared with those in model groups (t >or=2.76, P<0.05). With histological analysis, fibrotic scoring grade in octreotide-treated group was obviously ameliorated, compared with that in model groups (chi2 > or = 3.97, P<0.05). Imaging analysis revealed that alpha-SMA and TGFbeta1 immunohistological staining areas were markedly shrinked in octreotide-treated group (t > or = 2.47, P < 0.05). In two model groups, PCIII and type I mRNA levels significantly up-regulated as compared with those in normal group (t > or = 9.27, P<0.001), and they were inhibited by octreotide markedly (t > or = 2.47, P<0.05).</p><p><b>CONCLUSIONS</b>Octreotide can inhibit hepatic stellate cells transforming into myofibroblasts, down-regulate TGFbeta1, collagen type I and PCIII transcriptions, so that it has therapeutic effects on experimental hepatic fibrosis.</p>