[Effect of L-cysteine on colonic motility and the underlying mechanism].

The purpose of the present study is to investigate the effect of L-cysteine on colonic motility and the underlying mechanism. Immunohistochemical staining and Western blot were used to detect the localization of the H2S-generating enzymes cystathionine-ß-synthase (CBS) and cystathionine-γ-lyase (CSE). Organ bath system was used to observe the muscle contractile activities. Whole-cell patch-clamp technique was applied to record ionic channels currents in colonic smooth muscle cells. The results showed that both CBS and CSE were localized in mucosa, longitudinal and circular muscle and enteric neurons. L-cysteine had a dual effect on colonic contraction, and the excitatory effect was blocked by pretreatment with CBS inhibitor aminooxyacetate acid (AOAA) and CSE inhibitor propargylglycine (PAG); L-cysteine concentration-dependently inhibited L-type calcium channel current (ICa,L) without changing the characteristic of L-type calcium channel (P < 0.01); In contrast, the exogenous H2S donor NaHS increased ICa,L at concentration of 100 µmol/L, but inhibited ICa,L and modified the channel characteristics at concentration of 300 µmol/L (P < 0.05); Furthermore, L-cysteine had no effect on large conductance calcium channel current (IBKCa), but NaHS significantly inhibited IBKCa (P < 0.05). These results suggest that L-cysteine has a potential dual effect on colonic smooth muscle and the inhibitory effect might be directly mediated by L-type calcium channel while the excitatory effect might be mediated by endogenous H2S.

[IL-6 inhibits colonic longitudinal muscle contraction by inactivating L-type calcium channel in rats with pancreatitis].

The aim of this study was to investigate the effect of interleukin 6 (IL-6) on the contraction of colon longitudinal muscle strips in rats with acute pancreatitis (AP) and its underlying mechanism. Rat AP model was established by combined injection (i. p.) of ceruletide and lipopolysaccharide. The effect of IL-6 on spontaneous contraction of longitudinal smooth muscle strips of rat colon was observed by biological function experiment system. The level of serum IL-6 was detected by ELISA, the expression and distribution of IL-6 in colon were observed by histochemical staining, and the effect of IL-6 on L-type calcium channel in colon smooth muscle cells was observed by whole cell patch clamp technique. The results showed that, compared with the control group, AP group exhibited reduced contractile amplitude and longer contraction cycle of colon smooth muscle strips. IL-6 prolonged the contraction cycle of colon smooth muscle strips, but did not affect their spontaneous contraction amplitude. Serum IL-6 concentration in AP group was significantly higher than that in control group (P > 0.05). IL-6 was diffusely distributed in the colon of the control group, but the expression of IL-6 was significantly up-regulated in the colon gland, mucosa and submucosa of the AP group. IL-6 significantly decreased the peak current density of L-type calcium channel in rat colon smooth muscle cells. These results suggest that the colon motility of AP rats is weakened, and the mechanism may be that up-regulated IL-6 inactivates L-type voltage-dependent calcium channels, and then inhibits the contraction of colon longitudinal smooth muscle.

[Actions of endogenous hydrogen sulfide on colonic hypermotility in a rat model of chronic stress].

The present study was designed to investigate the potential role of endogenous hydrogen sulfide (H2S) in chronic stress-induced colonic hypermotility. Male Wistar rats were submitted daily to 1 h of water avoidance stress (WAS) or sham WAS (SWAS) for 10 consecutive days. The total number of fecal pellets was counted at the end of each 1 h of WAS or SWAS session. Organ bath recordings were used to test the colonic motility. H2S production of colon was determined, and immunohistochemistry and Western blot were performed on rat colonic samples to detect the distribution and expression of H2S-producing enzymes. The results showed that i) repeated WAS increased the number of fecal pellets per hour and the area under the curve (AUC) of the spontaneous contractions of colonic strips (P < 0.05), ii) repeated WAS decreased the endogenous production of H2S and the expression of H2S-producing enzymes in the colon devoid of mucosa and submucosa (P < 0.001), iii) cystathionine-γ-lyase (CSE) was strongly expressed in the cytosols of the circular and longitudinal smooth muscle cells and the nucleus of the myenteric plexus neurons, iv) cystathionine-ß-synthase (CBS) was primarily localized in the cytosols of myenteric plexus neurons and weakly localized in the epithelial cells and v) inhibitors of H2S-producing enzymes increased the contractile activity of colonic strips in the SWAS rats (P < 0.001). In conclusion, the results suggest that the colonic hypermotility induced by repeated WAS may be associated with the decreased production of endogenous H2S.

Heat shock protein 70 gene polymorphisms and cancer risk: a meta-analysis.

The polymorphisms in the three main heat shock protein 70 (HSP70-1, HSP70-2, and HSP70-hom) genes were identified to be associated with cancer risk. However, the results are inconsistent. We perform a meta-analysis to evaluate the association between the three HSP70 polymorphisms and cancer risk. Relevant studies were identified using PubMed, Web of Science, Chinese National Knowledge Infrastructure (CNKI), and Wanfang databases up to March 29, 2014. The cancer risk associated with the HSP70 polymorphisms was estimated for each study by odds ratios (OR) together with its 95% confidence interval (CI), respectively. Twenty case-control studies from eighteen publications were included; a significant association was observed for HSP70-2 polymorphism (dominant model: OR = 1.53, 95% CI: 1.11-2.09; recessive model: OR = 1.91, 95% CI: 1.06-3.45; AG versus AA: OR = 1.38, 95% CI: 1.03-1.84; GG versus AA: OR = 2.34, 95% CI: 1.21-4.54), while there was no significant association for HSP70-1 and HSP70-hom polymorphisms. Besides, in stratification analyses by ethnicity, cancer type, and source of control, significant association was detected for HSP70-2 polymorphism, while for HSP70-hom polymorphism, we found a significant association in hospital-based population under homozygote comparison model. This meta-analysis suggests that the HSP70-2 polymorphism rather than HSP70-hom and HSP70-1 polymorphisms was associated with the risk of cancer.

The DNA methyltransferase inhibitor zebularine induces mitochondria-mediated apoptosis in gastric cancer cells in vitro and in vivo.

DNA methyltransferase (DNMT) inhibitor zebularine has been reported to potentiate the anti-tumor effect by reactivating the expression of tumor suppressor genes and apoptosis-related genes in various malignant cells. However, the apoptotic signaling pathway in gastric cancer cells induced by zebularine is not well understood. In the study, the effects of zebularine on the growth and apoptosis of gastric cancer cells were investigated by MTT assay, Hoechst assay, Western blot analysis, flow cytometric analysis of annexin V-FITC/PI staining, and TUNEL assay. Zebularine was an effective inhibitor of human gastric cancer cells proliferation in vitro and in vivo. The effects were dose dependent. A zebularine concentration of 50 µM accounted for the inhibition of cell proliferation of 67% at 48 h. The treatment with zebularine upregulated Bax, and decreased Bcl-2 protein. Caspase-3 was activated, suggesting that the apoptosis is mediated by mitochondrial pathways. Moreover, zebularine injection successfully inhibited the tumor growth via apoptosis induction which was demonstrated by TUNEL assay in xenograft tumor mouse model. These results demonstrated that zebularine induced apoptosis in gastric cancer cells via mitochondrial pathways, and zebularine might become a therapeutic approach for the treatment of gastric cancer.

[Effects of methane on proximal colon motility of rats and ion channel mechanisms].

OBJECTIVE: To explore the effects of methane on proximal colon motility and elucidate its ion channel mechanisms. METHODS: The circular muscle and longitudinal muscle strips of proximal colon were isolated from rats. An isometric force transducer and a biology signal collection system were employed to observe the effects of methane on spontaneous contractile activity of muscle strips. The effects of methane on longitudinal muscle strips were observed after pre-treatment of these strips with tetrodotoxin (TTX) or N-nitro-L-arginine methylester (L-NAME). Single cell of colonic smooth muscle was isolated by collagenase and then whole-cell patch clamp technique was used to record voltage dependent potassium current (IKV) and large conductance Ca(2+)-activated K(+) current (IBKca) in the absence or presence of methane. RESULTS: Methane significantly attenuated the contractile amplitude of longitudinal muscle strips (from (1.12 ± 0.27) to (0.99 ± 0.31) g, n = 19, P = 0.013) whereas there were no changes in some longitudinal muscle strips (n = 6) in the presence of methane. The inhibitory effect of methane persisted after the pre-treatment of longitudinal muscle strips with TTX or L-NAME. Methane had no effects on the contractile cycle time of longitudinal muscle strips and contractile activity of circular muscle strips. And 3% methane solution significantly increased the density of IKV (from (13.3 ± 1.0) pA/pF to (18.5 ± 1.4) pA/pF, at +60 mV, n = 11, P = 0.001) versus the control group whereas methane had no effect on IBKca (all P > 0.05). CONCLUSION: Methane can inhibit contractile activity of proximal colonic longitudinal muscle by activating voltage dependent potassium channel and increasing IKV.

[Aspartate-ornithine granules in the treatment of nonalcoholic steatohepatitis: a multiple-dose parallel controlled clinical trial].

OBJECTIVE: To investigate the therapeutic efficacy and safety of aspartate-ornithine granules in patients with nonalcoholic steatohepatitis (NASH). METHODS: Seventy-two patients with NASH were included in this multiple-dose parallel controlled clinical trial and received a 12-week course of aspartate-ornithine granule treatment at either high-dose (6 g bid po; n = 38) or low-dose (3 g bid po; n = 34). Clinical efficacy was assessed by monitoring data from urinalysis, serologic tests (alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma-glutamyl transferase (GGT), and triglyceride (TG)), and abdominal computed tomography (CT) scan. Safety was assessed by occurrence of adverse events (fatigue, anorexia, abdominal distension, nausea, and vomiting). Statistical analyses were conducted to determine the significance of differences between parameters before (baseline) and after treatment. RESULTS: After 12 weeks of treatment, the liver and spleen CT ratios in both the high-dose group (0.89 +/- 0.19) and the low-dose group (0.80 +/- 0.15) were significantly higher than at baseline (S = 329, P less than 0.0001 and S = 246, P less than 0.0001); the overall improvement was more robust in the high-dose group (52.63%) than in the low-dose group (38.23%) (Z = -2.1042, P less than 0.05). After 6 and 12 weeks of treatment, the serum ALT levels in both the high-dose group and the low-dose group were significantly lower than at baseline (6 weeks: S = 324.5, P less than 0.0001 and S = 223, P less than 0.0001; 12 weeks: S = 370.5, P less than 0.0001 and S = 297.5, P less than 0.0001); the overall improvement was more robust in the high-dose group (79.0%) than in the low-dose group (53.0%) (Z = -2.0533, P less than 0.05). Similar trends were seen for the serum levels of AST and GGT after 6 and 12 weeks of treatment (all P less than 0.01) and serum levels of TG after 12 weeks of treatment. The rate of adverse reactions was low and similar between the two groups (high-dose: 4.8% and low-dose: 4.4%; all gastrointestinal). CONCLUSION: Aspartate-ornithine granule therapy was an effective and safe treatment of nonalcoholic steatohepatitis, with the higher dose of 6 g bid po providing more robust clinical benefit without affecting the safety profile.

Quantitative analysis of intestinal gas in patients with acute pancreatitis.

BACKGROUND: Disturbance of gastrointestinal function is a common complication in the early phase of acute pancreatitis (AP). Intestinal gas may reflect the function of the gut. Using plain abdominal radiographs, we investigated whether intestinal gas volume is related to AP. METHODS: Plain abdominal radiographs of 68 patients with AP within 24 hours after admission and 21 normal controls were digitized and transmitted to a computer. The region of intestinal gas was identified by an image manipulation software and the gas volume score (GVS) was calculated. The relationships between the GVS values and various clinical factors of AP were analyzed. RESULTS: The GVS in the AP group was 0.084+/-0.016, in the mild AP (MAP) group 0.070+/-0.005, and in the severe AP (SAP) group 0.094+/-0.013; all values were higher than that in the control group (P<0.01). The GVS in the SAP group was higher than that in the MAP group. The GVSs were correlated to the Ranson's scores (r=0.762, P<0.01) and the acute physiology and chronic health evaluation II (APACHE II) scores (r=0.801, P<0.01). In addition, the GVS in patients with secondary pancreatic and/or peripancreatic infection was 0.107+/-0.014, higher than that in patients without secondary infection (P<0.01). GVS was not related to gender, age, etiology or clinical outcome of AP. CONCLUSIONS: Intestinal gas volume is significantly elevated in patients with AP. It is closely related to Ranson's and APACHE II score and secondary pancreatic and/or peripancreatic infection. GVS may be a new prognostic tool for assessing the severity of AP in the early course of the disease.

[Clinical characteristics of ischemic bowel disease in young and middle-aged patients].

OBJECTIVE: To explore the clinical characteristics of ischemic bowel disease (IBD) in young and middle-aged patients (aged under 65 years old). METHODS: A total of 33 young and middle-aged IBD patients from January 1997 to July 2011 at Department of Gastroenterology, Renmin Hospital of Wuhan University were studied retrospectively. RESULTS: Among them, 4 patients underwent surgical procedures while 29 patients received conservative medical management. All became cured after correct treatment. The disease with a nighttime onset took up 36.4% (12/33). Among them, 21.2% (7/33) took oral contraceptives previously. Patients with no specific pre-existing condition accounted for 39.4% (13/33). Abdominal pain and hematochezia were the chief complaints of all patients. The laboratory findings were non-specific. The radiographic examination remained a major diagnostic tool. And 51.5% (17/33) patients had typical colonoscopic manifestations. CONCLUSION: More attention should be paid to young and middle-aged patients with classic abdominal pain, hematochezia and a lack of baseline cardiovascular disease.

[Role and mechanism of motilin in colonic motility disorder].

OBJECTIVE: To explore the role and mechanism of motilin in colonic motility disorder. METHODS: A total of 20 male Wistar rats (180 - 200 g) were randomly divided into 2 groups: water avoidance stress (WAS, n = 10) and sham water avoidance stress (SWAS, n = 10). Rats were exposed to 1 h WAS or SWAS daily for 10 consecutive days. Motilin in plasma was measured by enzyme-linked immunosorbent assay (ELISA). Proximal colon circular smooth muscle cells (PCSM) were isolated by enzymatic digestion and L-type calcium currents (ICa(L)) recorded by patch-clamp techniques. RESULTS: The fecal pellets during 1 h WAS significantly increased (5.4 ± 1.0 vs 2.4 ± 0.7, P < 0.01, n = 10). The motilin in plasma had significant difference between WAS rats and SWAS rats ((135 ± 35) vs (89 ± 24) pg/ml, P < 0.01, n = 10). The ICa(L) of two rats had no significant difference. But 6 × 10(-5) mmol/L motilin increased ICa(L) more in WAS than in SWAS rats at 0 mV ((1.6 ± 0.4) vs (1.0 ± 0.3) pA/pF, P < 0.05, n = 10). CONCLUSION: WAS leads to elevated motilin levels in plasma and active L-type Ca(2+) channels in colon. And it contributes to colonic motility disorder.

Comparison of high-dose dual therapy with bismuth-containing quadruple therapy in Helicobacter pylori-infected treatment-naive patients: An open-label, multicenter, randomized controlled trial.

OBJECTIVE: Bismuth-containing quadruple therapy for Helicobacter pylori (H. pylori) eradication has a relatively high rate of side effects and high cost, thus the option of a high-dose dual therapy with a high eradication rate and fewer adverse events is a consideration. However, studies of dual therapy are still scarce and are mostly single-center studies with limited generalizability. Large-scale, multicenter studies are required. Our study investigated and compared the effectiveness, adverse events, patient compliance, and costs of high-dose dual therapy with those of bismuth-containing quadruple therapy in H. pylori-infected treatment-naive patients in a prospective, multicenter, open-label, randomized controlled trial. METHOD: Treatment-naive patients infected with H. pylori were randomly assigned to receive high-dose dual therapy (esomeprazole 20 mg 4 times daily and amoxicillin 1000 mg 3 times daily, for 14 days) or bismuth-containing quadruple therapy (esomeprazole 20 mg, amoxicillin 1000 mg, clarithromycin 500 mg, and bismuth potassium citrate 220 mg, all twice daily for 14 days). The effectiveness, adverse events, patient compliance, and costs of both groups were compared. RESULTS: A total of 700 patients were enrolled. The high-dose dual therapy group (N = 350) achieved eradication rates of 89.4% (intention-to-treat), 90.4% (modified intention-to-treat), and 90.6% (per-protocol), which were similar to rates in the bismuth-containing quadruple therapy group (N = 350), 84.6%, 88.0%, and 88.2%, respectively (p > 0.05). The high-dose dual therapy group had a lower rate of adverse events (12.9% vs. 28.1%, p < 0.001) and lower costs (¥590.2 vs. ¥723.22) compared with the quadruple therapy group, respectively. The compliance of both groups was satisfactory (97.7% high-dose dual vs. 96.8% quadruple, p > 0.05). CONCLUSION: High-dose dual therapy for H. pylori eradication had similar efficacy and compliance, fewer adverse events, and lower costs than bismuth-containing quadruple therapy for treatment-naive patients.

Impaired activation of phosphatidylinositol 3-kinase by leptin is a novel mechanism of hepatic leptin resistance in NAFLD.

BACKGROUND/AIMS: The aim of this study was to detect the levels of leptin in serum and the expression of leptin, obesity receptor (OB-R), phosphatidylinositol 3-Kinase (p85) (PI3-K p85) and phospho-Akt-kinase (Akt) in non-alcoholic fatty liver disease (NAFLD). METHODOLOGY: The expressions of leptin, OB-R and PI3-K/ Akt kinase pathway were examined by immunohistochemistry. The level of leptin in serum was measured by radioimmunoassay. RESULTS: In agreement with significantly elevated serum leptin levels in NAFLD patients (p<0.05), expression of leptin, OB-R and PI3-K (p85) was significant higher in NAFLD patients (p<0.05) compared with the control patients. In contrast, expression of Akt was significantly down-regulated in the NAFLD patients (p<0.05). Moreover, PI3-K (p85) expression was significantly, positively correlated with leptin (r= 0.365, p<0.05) but negatively correlated with Akt (r=-0.854, p<0.01). CONCLUSIONS: Leptin may be involved in NAFLD pathogenesis by activating the PI3-K/Akt kinase pathway via OB-R and the defective leptin activation of PI3-K is a novel mechanism of leptin resistance in NAFLD.

[Effects of cholecystokinin octapeptide on the contractile activity of guinea-pig colonic smooth muscles, L-type calcium currents and membrane potentials of myocytes].

OBJECTIVE: To investigate the effects and mechanism of cholecystokinin octapeptide (CCK-8S) on the contractile activity of smooth muscles, L-type calcium current and membrane potentials of proximal colon myocytes in guinea pig. METHODS: (1) Strips of proximal colon were obtained from adult guinea pigs. The contraction of these stripes was measured by a RM6240 multi-channel physiological signal system. (2) Suspension of single smooth muscle cells (SMCs) were obtained from proximal colon and isolated by enzymatic digestion. The effect of CCK-8S on intracellular calcium concentration ([Ca(2+)]i) of SMCs was examined by fura-2-loaded microfluorimetric measurement. (3) Resting potential (RP), action potential (AP) and L-type calcium current (I(Ca-L)) were recorded by patch-clamp technique. RESULTS: (1) The contractile amplitude and frequency of muscle stripes enhanced by CCK-8S (10(-7) mol/L) were (149 ± 12)% and (132 ± 13)% respectively of those of control group (all P < 0.05). They were significantly attenuated by pretreating strips with CCK1 receptor antagonist devazepide (10(-7) mol/L), L-type calcium channel blocker nifedipine (10(-5) mol/L), Ca(2+)-ATPase inhibitor TG (thapsigargin) (10(-5) mol/L) and BA (boric acid) (10(-5) mol/L) respectively. (2) [Ca(2+)]i of SMCs intensified by CCK-8S was (738 ± 24)% of that of control group. And it was inhibited by pretreating SMCs with devazepide (all P < 0.05). (3) After the superfusion of CCK-8S, RP depolarized to (52 ± 9)%, the exogenously stimulated peak values of AP rose to (140 ± 4)% and fast repolarization time of AP decreased to (61 ± 13)% (all P < 0.05). They were significantly inhibited when these cells were pretreated with devazepide and/or nifedipine (n = 8, P < 0.05 for each group) whereas CI 988 had little effect. (4) The CCK-8S-evoked I(Ca-L) of SMCs at the voltage of + 10 mV was boosted to (138 ± 7)%. Such an effect was suppressed by a pretreatment with nifedipine, devazepide, TG and BA respectively. In the presence of an inhibitor of inositol 1,4,5-trisphosphate (IP3) receptors, heparin (10(-6) mol/L) and an protein kinase C (PKC) inhibitor, saurosporine (10(-6) mol/L), CCK-8S did not significantly intensify I(Ca-L) (all P > 0.05). CONCLUSION: CCK-8S promotes proximal colon contraction by CCK1 receptors through the activation of IP3-mediated PKC pathway.

Prognostic significance of hERG1 expression in gastric cancer.

Previous studies have demonstrated that human ether-à-go-go-related potassium channel (hERG1) is highly expressed in many tumor cell lines, as well as in primary human cancers, and, hence, have a critical role in cell cycle progress and proliferation. In this study, hERG1 expression was investigated in gastric cancer by immunohistochemistry and/or reverse transcription polymerase chain reaction (RT-PCR). It was discovered that hERG1, which was negatively expressed in surrounding non-tumor tissues, switched to aberrantly positive expression in gastric cancer. Statistically, there were significant differences in hERG1 protein expression according to factors such as serosal invasion, venous invasion, lymph node metastases, other organ metastases, and stage. The mean survival time for the hERG1-positive expression group was significantly shorter than the negative group, the survival rates for the positive group were significantly lower than the negative group, and hERG1 expression was found to be an independent prognostic factor. In summary, hERG1 channel was proved to be a potential biomarker for gastric cancer invasion and survival.

[The expression and role of PTEN in doxorubicin induced gastric cancer cell apoptosis].

OBJECTIVE: To study the expression of PTEN and its significance in doxorubicin-treated gastric cancer cells. METHODS: (1) Gastric cancer BGC-823 cells were treated with doxorubicin. Cell proliferation and apoptosis were evaluated by MTT and flow cytometry. The expression of PTEN at the mRNA and protein level were determined by RT-PCT and Western blot, respectively. (2) The gastric cancer xenografts model was constructed. The apoptosis of gastric cancer xenografts cells was determined by TUNEL. The expression of PTEN at the mRNA and protein level were detected using RT-PCR and Western blot, respectively. (3)BGC-823 cells were transfected with PTEN siRNA before addition of doxorubicin. The proliferation and apoptosis of these cells as well as the expression level of PTEN protein were determined. RESULTS: (1) After administration of doxorubicin, the proliferation of BGC-823 cells was inhibited in a time-dependent manner. (2) Doxorubicin significantly induced apoptosis of BGC-823 cells. (3) Doxorubicin treated BGC-823 cells showed a significant increase in the expression of PTEN at the mRNA and protein level in a time-dependent manner. TUNEL assay also showed a significant increase of apoptosis rate in gastric cancer xenografts treated with doxorubicin compared with control group [(28.11 ± 1.05)% vs (2.78 ± 1.63)%]. The expression of PTEN at the mRNA and protein level in the gastric cancer xenografts were significantly increased after administration of doxorubicin (0.5667 ± 0.0043 vs 0.2217 ± 0.0063, 0.14 ± 0.26 vs 0.04 ± 0.15, P < 0.05). (4) After treated with doxorubicin, the expression of PTEN in siRNA-transfected BGC-823 cells was significantly higher than that in non-transfected BGC-823 cells (P < 0.0001). The apoptosis of PTEN siRNA-transfected BGC-823 cells was significantly decreased compared with non-transfected BGC-823 cells [(10.35 ± 1.04)% vs (31.37 ± 3.58)%, P < 0.05]. CONCLUSION: Doxorubicin can effectively inhibit the growth and induce the apoptosis of BGC-823 gastric cancer cells. Increasing PTEN protein may be one of the main mechanism involved in this effect.

[Expression of Dickkopf-3 in esophageal squamous cell carcinoma].

OBJECTIVE: To investigate the expression of Dickkopf-3 (Dkk-3) in esophageal cancer and normal esophageal tissue and the relationship between Dkk-3 expression and the biological behavior of esophageal cancer. METHODS: Immunohistochemical method of S-P was used to examine Dkk-3 expression in 69 cases of esophageal squamous cell carcinoma and 5 cases of normal esophageal tissue with non-tumor tissue microarray and the results were analyzed and correlated with their clinical and pathological features. RESULTS: Positive Dkk-3 expression was observed in 65.7% (44/67) of the esophageal squamous cell carcinoma cases, but only one of the five cases with normal esophageal tissue showed positive microvascular expression of Dkk-3. In cases with positive expression of Dkk-3 significant differences were found in fiber membrane infiltration, depth of invasion, lymph node metastasis and TNM staging (P < 0.05), while no significant differences were found in the age, gender and pathological grading (P > 0.05). CONCLUSIONS: The upregulation of Dkk-3 expression in esophageal squamous cell carcinoma may contribute to tumor invasion and metastasis.

Clinicopathological association and prognostic value of long non-coding RNA CASC9 in patients with cancer: A meta-analysis.

Several studies have reported a prognostic role of the long non-coding RNA (lncRNA) cancer susceptibility candidate 9 (CASC9) in various cancer types, but its clinical significance has remained inconclusive. The aim of the present meta-analysis was to evaluate the impact of CASC9 expression on the prognosis and clinicopathological features of patients with cancer patients. The PubMed, Embase, Cochrane Library and Web of Science databases were searched for relevant literature and eight studies, including 565 patients with cancer, were selected. The quality of these studies was appraised with the Newcastle-Ottawa Scale (NOS) and the association between CASC9 expression and prognosis or clinicopathological features was analyzed. Patients with high expression levels of CASC9 in their tumor tissues had a lower overall survival rate compared with those in the low CASC9 expression group (hazard ratio=2.25, 95% CI: 1.60-3.17, P<0.001). Furthermore, elevated CASC9 expression was significantly associated with deeper tumor invasion [odds ratio (OR)=2.66, 95% CI: 1.72-4.14, P<0.001], poor tumor differentiation (OR=2.44, 95% CI: 1.24-4.78, P=0.009), lymph node metastasis (OR=3.42, 95% CI: 1.98-5.92, P<0.001) and advanced clinical stage (OR=3.21, 95% CI: 2.21-4.66, P<0.001). In conclusion, CASC9 is a promising biomarker for predicting the prognosis of cancer patients and should be validated in the clinic.

Expression of C-kit messenger ribonucleic acid and C-kit protein in the gallbladders in guinea pigs of high cholesterol diet.

The c-kit protooncogene receptor and its ligand-stem cell factor regulating the proliferation and survival of interstitial cells of Cajal (ICCs) have been described. The aim of this study was to determine the expression of c-kit mRNA and c-kit protein in the gallbladders in guinea pigs of high cholesterol diet (HCD). The gallbladder samples from 16 guinea pigs of HCD and from 16 guinea pigs of standard diet (StD) were used for this study. Expression of c-kit mRNA was detected by reverse transcription polymerase chain reaction (RT-PCR), and expression of c-kit protein was detected by Western blot analysis. Serum total cholesterol (TC) (39 +/- 6 vs. 109 +/- 20 mg/dl), low density lipoprotein (LDL) cholesterol (24 +/- 4 vs. 71 +/- 10 mg/dl), high density lipoprotein (HDL) cholesterol (2.4 +/- 0.4 vs. 7.0 +/- 1.6 mg/dl), and triglyceride (TG) (58 +/- 8 vs. 118 +/- 23 mg/dl) concentrations were significantly higher in the HCD group than in the StD group of guinea pigs (P < 0.001, respectively). Decreased expression of c-kit mRNA was demonstrated in the HCD group compared with the StD group. The ratio of c-kit mRNA and GAPDH was 0.56 +/- 0.09 in controls and 0.50 +/- 0.07 in the HCD group (P = 0.033). C-kit protein expression significantly declined in the HCD group. The mean value of optical density was 129 +/- 25 in the StD group and 103 +/- 19 in the HCD group (P = 0.0009). The data indicate that the expression of c-kit mRNA and c-kit protein significantly decreased in the gallbladders in guinea pigs of HCD.

Sensory denervation reduces visceral hypersensitivity in adult rats exposed to chronic unpredictable stress: evidences of neurogenic inflammation.

The purpose of this study is to provide evidence of neurogenic inflammation in chronic unpredictable stressed rats with the changes of visceral sensitivity, number of mast cells, and close proximity among mast cell-nerve-blood vessels. We found that (1) capsaicin denervation blocked stress-induced increase of visceral sensitivity, while doxantrazole presented a partial blocking; (2) capsaicin denervation blocked stress-induced enhancement of the proximity of mast cell-nerve fiber-blood vessels and blood vessel damage, while doxantrazole showed no effects on these; (3) doxantrazole blocked stress-induced increases of the MPO activity, the number and the degranulation of mast cells in the colon; (4) sensory denervation and doxantrazole had no effects on stress-induced behavioral inhibition. These results suggest that capsaicin-sensitive sensory fibers play a key role in stress-induced visceral hypersensitivity and the ultrastructural changes, mast cells play an important role in the generation of stress-induced colon inflammation.

[Altered expression of L-type calcium channel alpha1C and alpha1D subunits in colon of rats with diarrhea-predominant irritable bowel syndrome].

OBJECTIVE: To investigate the molecular identities of L-type calcium channel alpha1C subunit (Cav1.2) and alpha1D subunit (Cav1.3) responsible for motor dysfunction of diarrhea-predominant irritable bowel syndrome (D-IBS). METHODS: A total of 30 male Wistar rats were randomly divided into two groups. The traditional method for irritable bowel syndrome in a cold environment and intragastric administration of Folium Cassiae were combined to develop the D-IBS model. The fecal particles of rats and the water content in feces were measured. Then the expression of Cav1.2 and Cav1.3 mRNA was detected by reverse transcription polymerase chain reaction. The expressions of Cav1.2 and Cav1.3 were examined by immunohistochemistry in colonic tissues from D-IBS model rats and matched tissues. RESULTS: The fecal particles and the water content in feces of D-IBS model rats significantly increased as compared with the normal rats (6.8 +/- 1.4 vs 3.2 +/- 0.8, P = 0.032, 80% +/- 4% vs 47% +/- 5%, P = 0.018). Cav1.2 and Cav1.3 were positively expressed in colon of both model group and control group rats. The immunohistochemical scores of Cav1.2 and Cav1.3 expression increased in colon of D-IBS model rats as compared with those in normal control rats (3.43 +/- 0.92 vs 2.82 +/- 0.60, P = 0.034, 4.32 +/- 0.51 vs 3.75 +/- 1.05, P = 0.039). The immunohistochemical scores of Cav1.3 expression were significantly higher than Cav1.2 in colon of both two group rats (P = 0.003, 0.005). Similarly, the expression of Cav1.2 and Cav1.3 mRNA increased in colon of D-IBS model rats as compared with those in normal control rats (1.18 +/- 0.15 vs 1.06 +/- 0.12, P = 0.023, 1.32 +/- 0.13 vs 1.23 +/- 0.13, P = 0.033). The expression of Cav1.3 mRNA was significantly higher than Cav1.2 in colon of both two group rats (P = 0.038, 0.012). CONCLUSIONS: The traditional modeling of irritable bowel syndrome in rats alters the expression of Cav1.2 and Cav1.3. It may be directly related to the generation of enhanced colonic contraction in D-IBS. In addition, Cav1.3 may play a more important role than Cav1.2 in colonic contractility. It offers another potential therapeutic target in the treatment of irritable bowel syndrome.