Melittin induces human gastric cancer cell apoptosis via activation of mitochondrial pathway

AIM: To investigate the apoptotic effects of melittin on SGC-7901 cells via activation of the mitochondrial signaling pathway in vitro.METHODS: SGC-7901 cells were stimulated by melittin, and its effect on proliferation and apoptosis of was investigated by methyl thiazolyl tetrazolium assay, morphologic structure with transmission electron microscopy, annexin-V/propidium iodide double-staining assay, measuring mitochondrial membrane potential(MMP) levels, and analyzing reactive oxygen species(ROS) concentrations were analyzed by flow cytometry. Cytochrome C(Cyt C), apoptosis-inducing factor(AIF), endonuclease G(Endo G), second mitochondria-derived activator of caspases(Smac)/direct IAP binding protein with low isoelectric point(Diablo), and FAS were analyzed by western blot. The expression of caspase-3 and caspase-8 was measured using activity assay kits.RESULTS: Melittin was incubated at 1.0, 2.0, 4.0, or 6.0 μg/m L for 1, 2, 4, 6, or 8 h and showed a timeand concentration-dependent inhibition of SGC-7901 cell growth. Melittin induced SGC-7901 cell apoptosis, which was confirmed by typical morphological changes. Treatment with 4 μg/m L melittin induced early apoptosis of SGC-7901 cells, and the early apoptosis rates were 39.97% ± 3.19%, 59.27% ± 3.94%, and 71.50% ± 2.87% vs 32.63% ± 2.75% for 1, 2, and 4 h vs 0 h(n = 3, P 0.05); the ROS levels were 616.53% ± 79.78%, 974.81% ± 102.40%, and 1330.94% ± 93.09% vs 603.74% ± 71.99%(n = 3, P 0.05); the MMP values were 2.07 ± 0.05, 1.78 ± 0.29, and 1.16 ± 0.25 vs 2.55 ± 0.42(n = 3, P 0.05); caspase-3 activity was significantly higher compared to the control(5492.3 ± 321.1, 6562.0 ± 381.3, and 8695.7 ± 449.1 vs 2330.0 ± 121.9), but the caspase activity of the non-tumor cell line L-O2 was not different from that of the control. With the addition of the caspase-3 inhibitor(Ac-DEVD-CHO), caspase-3 activity was significantly decreased compared to the control group(1067.0 ± 132.5 U/g vs 8695.7 ± 449.1 U/g). The expression of the Cyt C, Endo G, and AIF proteins in SGC-7901 cells was significantly higher than those in the control(P 0.05), while the expression of the Smac/Diablo protein was significantly lower than the control group after melittin exposure(P 0.01). Ac-DEVD-CHO did not, however, have any effect on the expression of caspase-8 and FAS in the SGC-7901 cells.CONCLUSION: Melittin can induce apoptosis of human gastric cancer(GC) cells through the mitochondria pathways, and it may be a potent agent in the treatment of human GC.     

Function of chloride intracellular channel 1 in gastric cancer cells

AIM: To investigate the effect of chloride intracellular channel 1 (CLIC1) on the cell proliferation, apoptosis, migration and invasion of gastric cancer cells.METHODS: CLIC1 expression was evaluated in human gastric cancer cell lines SGC-7901 and MGC-803 by real time polymerase chain reaction (RT-PCR). Four segments of small interference RNA (siRNA) targeting CLIC1 mRNA and a no-sense control segment were designed by bioinformatics technology. CLIC1 siRNA was selected using Lipofectamine 2000 and transfected transiently into human gastric cancer SGC-7901 and MGC-803 cells. The transfected efficiency was observed under fluorescence microscope. After transfection, mRNA expression of CLIC1 was detected with RT-PCR and Western blotting was used to detect the protein expression. Proliferation was examined by methyl thiazolyl tetrazolium and apoptosis was detected with flow cytometry. Polycarbonate membrane transwell chamber and Matrigel were used for the detection of the changes of invasion and migration of the two cell lines.RESULTS: In gastric cancer cell lines SGC-7901 and MGC-803, CLIC1 was obviously expressed and CLIC1 siRNA could effectively suppress the expression of CLIC1 protein and mRNA. Proliferation of cells transfected with CLIC1 siRNA3 was enhanced notably, and the highest proliferation rate was 23.3% (P = 0.002) in SGC-7901 and 35.55% (P = 0.001) in MGC-803 cells at 48 h. The G2/M phase proportion increased, while G0/G1 and S phase proportions decreased. The apoptotic rate of the CLIC1 siRNA3 group obviously decreased in both SGC-7901 cells (62.24%, P = 0.000) and MGC-803 cells (52.67%, P = 0.004). Down-regulation of CLIC1 led to the inhibition of invasion and migration by 54.31% (P = 0.000) and 33.62% (P = 0.001) in SGC-7901 and 40.74% (P = 0.000) and 29.26% (P = 0.002) in MGC-803. However, there was no significant difference between the mock group cells and the negative control group cells.CONCLUSION: High CLIC1 expression can efficiently inhibit proliferation and enhance apoptosis, migration and invasion of gastric cancer cells in vitro. CLIC1 might be a promising target for the treatment of gastric cancer.     

Cortactin expression confers a more malignant phenotype to gastric cancer SGC-7901 cells

AIM:To study the effects of cortactin on the tumor biology of SGC-7901 cells and identify the mechanism involved in the process.METHODS:Cell lines in which cortactin was stably overexpressed or knocked down as well as the respective control cell lines were established by standard molecular methods.The effects of cortactin on the proliferation,migration and invasion capacity of SGC-7901cells were assessed by the MTT assay,colony formation,flow cytometry,transwell migration and matrigel invasion.Nude mouse models were also used to assess the role of cortactin in the growth and metastasis of SGC-7901 cells in vivo.Western blotting analysis was performed to detect the expression of epidermal growth factor receptor(EGFR)and downstream molecules.RESULTS:Cell lines in which cortactin was stably overexpressed or knocked down as well as control cell lines were successfully established and designated as LV5-cortactin-SGC,LV5-SGC,LV3-shRNA-SGC and LV3-SGC.Cortactin overexpression promoted SGC-7901 cell migration(340.7±12.6 vs 229.1±23.2,P<0.01)and invasion(71.6±5.2 vs 48.4±3.6,P<0.01).Cortactin downregulation impaired SGC-7901 cell migration(136.2±19.8 vs 225±17)and invasion(29.2±5.2vs 49.6±3.8,P<0.01).The results from the MTT and colony formation assays results indicated increased LV5-cortactin-SGC cell proliferation and decreased LV3-shRNA-SGC cell proliferation compared to the control cells.Flow cytometry analysis demonstrated that cortactin overexpression promoted the proliferation index of SGC-7901 cells,and the results were reversed when cortactin was downregulated.Mouse tumor models confirmed that cortactin expression increased SGC-7901cell proliferation and metastasis in vivo.Western blotting analysis revealed that cortactin elevated EGFR expression and activated the downstream molecules.CONCLUSION:Cortactin expression promoted the migration,invasion and proliferation of SGC-7901 cells both in vivo and in vitro.The EGFR signaling pathway is mechanistically involved.     

High dose glargine alters the expression profiles of microRNAs in pancreatic cancer cells

AIM: To investigate the effect of high dose glargine on the expression profiles of microRNAs in human pancreatic cancer cells.METHODS: Real-time polymerase chain reaction array (RT-PCR) was applied to investigate miRNAs differentially expressed in Sw1990 cells treated with or without 100 IU/L glargine. Stem-loop RT-PCR was used to confirm the results of the array assay in Sw1990 and Panc-1 cells. The effects of miR-95 on cell growth, apoptosis, invasion and migration abilities were respectively examined by CCK8 assay, apoptosis assay, Matrigel invasion and migration assay in Sw1990 and Panc-1 cells. Nude mice xenograft models with Sw1990 cells were built to investigate pancreatic cancer growth in vivo after transfection by the lentivirus pGLV3-GFP- miR-95.RESULTS: Ten miRNAs were significantly up-regulated and 2 miRNAs down-regulated in glargine treated Sw1990 cells when compared with non-treated cells (2.48-fold changes on average, P < 0.01). miR-95, miR-134 and miR-34c-3p are the top three miRNAs regulated by glargine (3.65-fold, 2.67-fold and 2.60-fold changes respectively, P < 0.01) in Sw1990 cells. Stem-loop RT-PCR confirmed that high dose glargine up-regulated the expression of miR-95 and miR-134 in both Sw1990 and Panc-1 cells. The most obvious change is the apparent increase of miR-95. Forced expression of miR-95 significantly increased cell proliferation (Sw1990: 2.510 ± 0.129 vs 2.305 ± 0.187, P < 0.05; Panc-1: 2.439 ± 0.211 vs 2.264 ± 0.117, P < 0.05), invasion (Sw1990: 67.90 ± 12.33 vs 47.30 ± 5.89, P < 0.01; Panc-1: 37.80 ± 8.93 vs 30.20 ± 5.14, P < 0.01), migration (Sw1990: 101 ± 6.00 vs 51.20 ± 8.34, P < 0.01; Panc-1: 91.80 ± 9.22 vs 81.50 ± 7.47, P < 0.01) and inhibited cell apoptosis (Sw1990: 22.05% ± 1.92% vs 40.32% ± 1.93%, P < 0.05; Panc-1: 20.17% ± 0.85% vs 45.60% ± 1.43%, P < 0.05) when compared with paired negative controls, whereas knockdown of miR-95 obtained the opposite effect. Nude mice xenograft models confirmed that miR-95 promoted the growth of pancreatic cancer in vivo when compared with negative control (tumor volume: 373.82 ± 23.67 mL vs 219.69 ± 17.82 mL, P < 0.05).CONCLUSION: These observations suggested that modulation of miRNA expression may be an important mechanism underlying the biological effects of glargine.     

Reduced expression of circulating microRNA-218 in gastric cancer and correlation with tumor invasion and prognosis

AIM:To investigate the expression of microRNA-218(miR-218)in serum from gastric cancer patients and its relationship with clinicopathological characteristics.METHODS:A total of 68 patients with pathologically diagnosed gastric cancer and 56 healthy individuals were recruited to this study.The expression of miR-218was detected in the serum of gastric cancer patients and healthy individuals by quantitative real-time polymerase chain reaction.The clinical data were collectedand analyzed by statistical software.RESULTS:miR-218 was reduced significantly in the serum of gastric cancer patients compared to healthy individuals(1.15±0.08 vs 0.37±0.023;P=0.026).In the gastric cancer group,serum expression of miR-218was lower in patients with metastasis and poorly differentiated cancer compared with non-metastatic and well-differentiated cancer(0.19±0.011 vs 0.45±0.021,P=0.031 and 0.21±0.019 vs 0.49±0.021,P=0.025).Serum miR-218 was found to be significantly associated with gastric cancer metastasis(P=0.003),tumor T stage(P=0.018)and tumor grade(P=0.012).Low serum expression of miR-218 was related to an increase in the stage of gastric cancer.The expression level of miR-218 in the serum was correlated with the3-year survival.Ninety-seven percent of patients with a high level of miR-218 expression survived for 3 years,while only 54%of those with low miR-218 expression survived.CONCLUSION:miR-218 is deregulated in gastric cancer patients and is strongly correlated with tumor stage,grade and metastasis.Serum expression of miR-218 may be a prognostic marker.     

Overexpressed miRNA-155 dysregulates intestinal epithelial apical junctional complex in severe acute pancreatitis

AIM:To investigate whether miRNA-155(miR-155)dysregulates apical junctional complex(AJC)protein expression in experimental severe acute pancreatitis(SAP).METHODS:Twenty-four male BALB/c mice were randomly assigned to two groups:the SAP group(n=12)receiving sequential intraperitoneal injection of 50μg/kg caerulein and 10 mg/kg lipopolysaccharide over 6h,and the control group(n=12)receiving intraperitoneal injection of normal saline.Animals were sacrificed3 h following the last injection for collection of blood samples and pancreas and distal ileal segment specimens.Routine pancreas and intestine histology was used to assess SAP pathology and intestinal epithelial barrier damage.Levels of serum amylase,diamine oxidase(DAO),and tumor necrosis factor(TNF)-αwere determined using commercial kits.Total RNA samples were isolated from intestinal epithelial specimens and reversely transcribed into cDNA.miR-155 and RhoA mRNA expression profiles were determined using quantitative real-time polymerase chain reaction.Target genes for miR-155 were predicted using the miRTarBase database,RNA22 and PicTar computational methods.Western blotting was performed to quantitate the protein expression levels of the target gene RhoA,as well as zonula occludens(ZO)-1 and E-cadherin,two AJC component proteins.RESULTS:Intraperitoneal injection of caerulein and lipopolysaccharide successfully induced experimental acute pancreatic damage(SAP vs control,10.0±2.0vs 3.2±1.2,P<0.01)and intestinal epithelial barrier damage(3.2±0.7 vs 1.4±0.7,P<0.01).Levels of serum amylase(21.6±5.1 U/mL vs 14.3±4.2 U/mL,P<0.01),DAO(21.4±4.1 mg/mL vs 2.6±0.8 mg/mL,P<0.01),and TNF-α(61.0±15.1 ng/mL vs 42.9±13.9 ng/mL,P<0.01)increased significantly in SAP mice compared to those in control mice.miR-155 was significantly overexpressed in SAP intestinal epithelia(1.94±0.50 fold vs 1.03±0.23 fold,P<0.01),and RhoA gene containing three miR-155-specific binding sites in the three prime untranslated regions was one of the target genes for mi     

Comparision of modified and conventional delta-shaped gastroduodenostomy in totally laparoscopic surgery

AIM:To evaluate the safety and feasibility of a modified delta-shaped gastroduodenostomy(DSG)in totally laparoscopic distal gastrectomy(TLDG).METHODS:We performed a case-control study enrolling 63 patients with distal gastric cancer(GC)undergoing TLDG with a DSG from January 2013 to June 2013.Twenty-two patients underwent a conventional DSG(Con-Group),whereas the other 41 patients underwent a modified version of the DSG(Mod-Group).The modified procedure required only the instruments of the surgeon and assistant to complete the involution of the common stab incision and to completely resect the duodenal cutting edge,resulting in an anastomosis with an inverted T-shaped appearance.The clinicopathological characteristics,surgical outcomes,anastomosis time and complications of the two groups were retrospectively analyzed using a prospectively maintained comprehensive database.RESULTS:DSG procedures were successfully completed in all of the patients with histologically complete(R0)resections,and none of these patients required conversion to open surgery.The clinicopathological characteristics of the two groups were similar.There were no significant differences between the groups in the operative time,intraoperative blood loss,extension of the lymph node(LN)dissection and number of dissected LNs(150.8±21.6 min vs 143.4±23.4 min,P=0.225for the operative time;26.8±11.3 min vs 30.6±14.8mL,P=0.157 for the intraoperative blood loss;4/18 vs3/38,P=0.375 for the extension of the LN dissection;and 43.9±13.4 vs 39.5±11.5 per case,P=0.151 for the number of dissected LNs).The anastomosis time,however,was significantly shorter in the Mod-Group than in the Con-Group(13.9±2.8 min vs 23.9±5.6min,P=0.000).The postoperative outcomes,including the times to out-of-bed activities,first flatus,resumption of soft diet and postoperative hospital stay,as well as the anastomosis size,did not differ significantly(1.9±0.6 d vs 2.3±1.5 d,P=0.228 for the time to outof-bed activities;3.2±0.9 d vs 3.5±1.3 d,P=0.295for the first flatus time;7.5±0.8 d vs 8.1±4.3 d,P=0.489 for the resumption of a soft diet time;14.3±10.6 d vs 11.5±4.9 d,P=0.148 for the postoperative hospital stay;and 30.5±3.6 mm vs 30.1±4.0 mm,P=0.730 for the anastomosis size).One patient with minor anastomotic leakage in the Con-Group was managed conservatively;no other patients experienced any complications around the anastomosis.The operative complication rates were similar in the Con-and ModGroups(9.1%vs 7.3%,P=1.000).CONCLUSION:The modified DSG,an alternative reconstruction in TLDG for GC,is technically safe and feasible,with a simpler process that reduces the anastomosis time.     

Axl glycosylation mediates tumor cell proliferation, invasion and lymphatic metastasis in murine hepatocellular carcinoma

AIM: To investigate the effects of Axl deglycosylation on tumor lymphatic metastases in mouse hepatocellular carcinoma cell lines. METHODS: Western blotting was used to analyze the expression profile of Axl glycoprotein in mouse hepa-tocellular carcinoma cell line Hca-F treated with tunicamycin and PNGase F 3-(4,5)-dimethylthiazol(-zyl)-3,5- diphenyltetrazolium bromide (MTT) assay, extracellular matrix (ECM) invasion assay (in vitro ) and tumor metastasis assay (in vivo ) were utilized to evaluate the effect of Axl deglycosylation on the Hca-F cell proliferation, invasion and lymphatic metastasis. RESULTS: Tunicamycin and PNGase F treatment markedly inhibited Axl glycoprotein synthesis and expression, proliferation, invasion, and lymphatic metastasis both in vitro and in vivo . In the MTT assay, proliferation was apparent in untreated Hca-F cells compared with treated Hca-F cells. In the ECM invasion assay (in vitro ), treated cells passed through the ECMatrix gel in significantly smaller numbers than untreated cells (tunicamycin 5 μg/mL: 68 ± 8 vs 80 ± 9, P=0.0222; 10 μg/mL: 50 ± 6vs 80 ± 9,P=0.0003; 20 μg/mL: 41 ± 4 vs 80 ± 9, P=0.0001); (PNGase F 8 h: 66 ± 7 vs 82 ± 8, P=0.0098; 16 h: 49 ± 4 vs 82 ± 8, P=0.0001; 24 h: 34 ± 3 vs 82 ± 8, P=0.0001). In the tumor metastasis assay (in vivo ), average lymph node weights of the untreated Hca-F group compared with treated Hca-F groups (tunicamycin 5 μg/mL: 0.84 ± 0.21 g vs 0.72 ± 0.19 g, P=0.3237; 10 μg/mL: 0.84 ± 0.21 g vs 0.54 ± 0.11 g, P=0.0113; 20 μg/mL: 0.84 ± 0.21 g vs 0.42 ± 0.06 g, P=0.0008); (PNGase F 8 h: 0.79 ± 0.15 g vs 0.63 ± 0.13 g, P=0.0766; 16 h: 0.79 ± 0.15 g vs 0.49 ± 0.10 g, P=0.0022; 24 h: 0.79 ± 0.15 g vs 0.39 ± 0.05 g, P=0.0001). Also, average lymph node volumes of the untreated Hca-F group compared with treated Hca-F groups (tunicamycin 5 μg/mL: 815 ± 61 mm 3 vs 680 ± 59 mm 3 , P=0.0613; 10 μg/mL: 815 ± 61 mm 3 vs 580 ± 29 mm 3 , P=0.0001; 20 μg/mL: 815 ± 61 mm 3 vs 395 ± 12 mm 3 , P=0.0001); (PNGase F 8 h: 670 ± 56 mm 3 vs 581 ± 48 mm 3 , P=0.0532; 16 h: 670 ± 56 mm 3 vs 412 ± 22 mm 3 , P=0.0001; 24 h: 670 ± 56 mm 3 vs 323 ± 11 mm 3 , P=0.0001). CONCLUSION: Alteration of Axl glycosylation can at-tenuate neoplastic lymphatic metastasis. Axl N-glycans may be a universal target for chemotherapy.     

Prevalence of functional dyspepsia and its subgroups in patients with eating disorders

AIM: To study the prevalence of functional dyspepsia (FD) (Rome III criteria) across eating disorders (ED), obese patients, constitutional thinner and healthy volunteers.METHODS: Twenty patients affected by anorexia nervosa, 6 affected by bulimia nervosa, 10 affected by ED not otherwise specified according to diagnostic and statistical manual of mental disorders, 4th edition, nine constitutional thinner subjects and, thirty-two obese patients were recruited from an outpatients clinic devoted to eating behavior disorders. Twenty-two healthy volunteers matched for age and gender were enrolled as healthy controls. All participants underwent a careful clinical examination. Demographic and anthropometric characteristics were obtained from a structured questionnaires. The presence of FD and, its subgroups, epigastric pain syndrome and postprandial distress syndrome (PDS) were diagnosed according to Rome III criteria. The intensity-frequency score of broader dyspeptic symptoms such as early satiety, epigastric fullness, epigastric pain, epigastric burning, epigastric pressure, belching, nausea and vomiting were studied by a standardized questionnaire (0-6). Analysis of variance and post-hoc Sheffè tests were used for comparisons.RESULTS: 90% of patients affected by anorexia nervosa, 83.3% of patients affected by bulimia nervosa, 90% of patients affected by ED not otherwise specified, 55.6% of constitutionally thin subjects and 18.2% healthy volunteers met the Postprandial Distress Syndrome Criteria (χ², P < 0.001). Only one bulimic patient met the epigastric pain syndrome diagnosis. Postprandial fullness intensity-frequency score was significantly higher in anorexia nervosa, bulimia nervosa and ED not otherwise specified groups compared to the score calculated in the constitutional thinner group (4.15 ± 2.08 vs 1.44 ± 2.35, P = 0.003; 5.00 ± 2.45 vs 1.44 ± 2.35, P = 0.003; 4.10 ± 2.23 vs 1.44 ± 2.35, P = 0.002, respectively), the obese group (4.15 ± 2.08 vs 0.00 ± 0.00, P < 0.001; 5.00 ± 2.45 vs 0.00 ± 0.00, P < 0.001; 4.10 ± 2.23 vs 0.00 ± 0.00, P < 0.001, respectively) and healthy volunteers (4.15 ± 2.08 vs 0.36 ± 0.79, P < 0.001; 5.00 ± 2.45 vs 0.36 ± 0.79, P < 0.001; 4.10 ± 2.23 vs 0.36 ± 0.79, P < 0.001, respectively). Early satiety intensity-frequency score was prominent in anorectic patients compared to bulimic patients (3.85 ± 2.23 vs 1.17 ± 1.83, P = 0.015), obese patients (3.85 ± 2.23 vs 0.00 ± 0.00, P < 0.001) and healthy volunteers (3.85 ± 2.23 vs 0.05 ± 0.21, P < 0.001). Nausea and epigastric pressure were increased in bulimic and ED not otherwise specified patients. Specifically, nausea intensity-frequency-score was significantly higher in bulimia nervosa and ED not otherwise specified patients compared to anorectic patients (3.17 ± 2.56 vs 0.89 ± 1.66, P = 0.04; 2.70 ± 2.91 vs 0.89 ± 1.66, P = 0.05, respectively), constitutional thinner subjects (3.17 ± 2.56 vs 0.00 ± 0.00, P = 0.004; 2.70 ± 2.91 vs 0.00 ± 0.00, P = 0.005, respectively), obese patients (3.17 ± 2.56 vs 0.00 ± 0.00, P < 0.001; 3.17 ± 2.56 vs 0.00 ± 0.00, P < 0.001 respectively) and, healthy volunteers (3.17 ± 2.56 vs 0.17 ± 0.71, P = 0.002; 3.17 ± 2.56 vs 0.17 ± 0.71, P = 0.001, respectively). Epigastric pressure intensity-frequency score was significantly higher in bulimic and ED not otherwise specified patients compared to constitutional thin subjects (4.67 ± 2.42 vs 1.22 ± 1.72, P = 0.03; 4.20 ± 2.21 vs 1.22 ± 1.72, P = 0.03, respectively), obese patients (4.67 ± 2.42 vs 0.75 ± 1.32, P = 0.001; 4.20 ± 2.21 vs 0.75 ± 1.32, P < 0.001, respectively) and, healthy volunteers (4.67 ± 2.42 vs 0.67 ± 1.46, P = 0.001; 4.20 ± 2.21 vs 0.67 ± 1.46, P = 0.001, respectively). Vomiting was referred in 100% of bulimia nervosa patients, in 20% of ED not otherwise specified patients, in 15% of anorexia nervosa patients, in 22% of constitutional thinner subjects, and, in 5.6% healthy volunteers (χ², P < 0.001).CONCLUSION: PDS is common in eating disorders. Is it mandatory in outpatient gastroenterological clinics to investigate eating disorders in patients with PDS?     

Health-related quality of life of 256 recipients after liver transplantation

AIM:To investigate health-related quality of life(HRQoL) and psychological outcomes in 256 adults who had undergone liver transplantation(LT).METHODS:A stratified random sampling method was used in this follow-up multicenter study to select a representative sample of recipients undergoing either living donor liver transplantation(LDLT) or deceased donor liver transplantation(DDLT).HRQoL was measured by using the Chinese version of Medical Outcome Study Short Form-36(SF-36),and psychological outcomes by using the beck anxiety inventory(BAI) and the self-rating depression scale(SDS).Clinical and demographic data were collected from the records of the Chinese Liver Transplant Registry and via questionnaires.RESULTS:A total of 256 patients were sampled,including 66(25.8%) receiving LDLT and 190(74.2%) undergoing DDLT;15(5.9%) recipients had anxiety and four(1.6%) developed severe depression after the operation.Compared with LDLT recipients,DDLT patients had higher scores in general health(60.33 ± 16.97 vs 66.86 ± 18.42,P = 0.012),role-physical(63.64 ± 42.55 vs 74.47 ± 36.46,P = 0.048),roleemotional(61.11 ± 44.37 vs 78.95 ± 34.31,P = 0.001),social functioning(78.60 ± 22.76 vs 88.16 ± 21.85,P = 0.003),vitality(70.30 ± 15.76 vs 75.95 ± 16.40,P = 0.016),mental health(65.88 ± 12.94 vs 71.85 ± 15.45,P = 0.005),physical component summary scale(PCS,60.07 ± 7.36 vs 62.58 ± 6.88,P = 0.013) and mental component summary scale(MCS,52.65 ± 7.66 vs 55.95 ± 10.14,P = 0.016).Recipients 45 years old at the time of transplant scored higher in vitality(77.33 ± 15.64 vs 72.52 ± 16.66,P = 0.020),mental health(73.64 ± 15.06 vs 68.00 ± 14.65,P = 0.003) and MCS(56.61 ± 10.00 vs 54.05 ± 9.30,P = 0.037) than those aged ≤ 45 years.MCS was poorer in recipients with than in those without complications(52.92 ± 12.21 vs 56.06 ± 8.16,P = 0.017).Regarding MCS(55.10 ± 9.66 vs 50.0 ± 10.0,P 0.05) and PCS(61.93 ± 7.08 vs 50.0 ± 10.0,P 0.05),recipients scored better than the Sichuan general and had improved overall QoL compared to patients with chronic diseases.MCS and PCS significantly correlated with scores of the BAI(P 0.001) and the SDS(P 0.001).CONCLUSION:Age 45 years at time of transplant,DDLT,full-time working,no complications,anxiety and depression were possible factors influencing postoperative HRQoL in liver recipients.     

Effects of hexahydrocurcumin in combination with 5-fluorouracil on dimethylhydrazine-induced colon cancer in rats

AIM: To investigate the effects of hexahydrocurcumin (HHC), and its combination with 5-fluorouracil (5-FU) on dimethylhydrazine (DMH)-induced colon cancer in rats.METHODS: Male Wistar rats weighing 100-120 g were used as subject models. Aberrant crypt foci (ACF), early preneoplastic lesions of colon cancer, were induced by subcutaneous injection of DHM (40 mg/kg) twice a week for two weeks. After the first DMH injection, rats were treated daily with vehicle (n = 12), curcumin (CUR) (50 mg/kg) (n = 12), HHC (50 mg/kg) orally (n = 12), and treated weekly with an intraperitoneal injection of 5-FU (50 mg/kg) (n = 12), or a combination of 5-FU plus CUR (n = 12) and HHC (n = 12) at the same dosage(s) for 16 wk. The total number of ACF and large ACF were assessed. Cyclooxygenase (COX)-1 and COX-2 expression were detected by immunohistochemistry in colon tissues. The quantitative data of both COX-1 and COX-2 expression were presented as the percentage of number of positive-stained cells to the total number of cells counted. Apoptotic cells in colon tissues were also visualized using the dUTP-biotin nick end labeling method. Apoptotic index (AI) was determined as the percentage of labeled nuclei with respect to the total number of nuclei counted.RESULTS: The total number of ACF was highest in the DMH-vehicle group (1558.20 ± 17.37), however, the number of ACF was significantly reduced by all treatments, 5-FU (1231.20 ± 25.62 vs 1558.20 ± 17.37, P < 0.001), CUR (1284.20 ± 25.47 vs 1558.20 ± 17.37, P < 0.001), HHC (1086.80 ± 53.47 vs 1558.20 ± 17.37, P < 0.001), DMH-5-FU + CUR (880.20 ± 13.67 vs 1558.20 ± 17.37, P < 0.001) and DMH-5-FU + HHC (665.80 ± 16.64 vs 1558.20 ± 17.37, P < 0.001). Interestingly, the total number of ACF in the combined treatment groups, the DMH-5-FU + CUR group (880.20 ± 13.67 vs 1231.20 ± 25.62, P < 0.001; 880.20 ± 13.67 vs 1284.20 ± 25.47, P < 0.001) and the DMH-5-FU + HHC group (665.80 ± 16.64 vs 1231.20 ± 25.62, P < 0.001; 665.80 ± 16.64 vs 1086.80 ± 53.47, P < 0.001) were significantly reduced as compared to 5-FU or each treatment alone. Large ACF were also significantly reduced in all treatment groups, 5-FU (111.00 ± 7.88 vs 262.20 ± 10.18, P < 0.001), CUR (178.00 ± 7.33 vs 262.20 ± 10.18, P < 0.001), HHC (186.60 ± 21.51 vs 262.20 ± 10.18, P < 0.001), DMH-5-FU + CUR (122.00 ± 5.94 vs 262.20 ± 10.18, P < 0.001) and DMH-5-FU + HHC (119.00 ± 17.92 vs 262.20 ± 10.18, P < 0.001) when compared to the vehicle group. Furthermore, in the DMH-5-FU + CUR and DMH-5-FU + HHC groups the formation of large ACF was significantly reduced when compared to CUR (122.00 ± 5.94 vs 178.00 ± 7.33, P < 0.005) or HHC treatment alone (119.00 ± 17.92 vs 186.60 ± 21.51, P < 0.001), however, this reduction was not statistically different to 5-FU monotherapy (122.00 ± 5.94 vs 111.00 ± 7.88, P = 0.217; 119.00 ± 17.92 vs 111.00 ± 7.88, P = 0.619, respectively). The levels of COX-1 protein after all treatments were not different from normal rats. A marked increase in the expression of COX-2 protein was observed in the DMH-vehicle group. Over-expression of COX-2 was not significantly decreased by 5-FU treatment alone (95.79 ± 1.60 vs 100 ± 0.00, P = 0.198). However, over-expression of COX-2 was significantly suppressed by CUR (77.52 ± 1.68 vs 100 ± 0.00, P < 0.001), HHC (71.33 ± 3.01 vs 100 ± 0.00, P < 0.001), 5-FU + CUR (76.25 ± 3.32 vs 100 ± 0.00, P < 0.001) and 5-FU + HHC (68.48 ± 2.24 vs 100 ± 0.00, P < 0.001) in the treated groups compared to the vehicle group. Moreover, CUR (77.52 ± 1.68 vs 95.79 ± 1.60, P < 0.001), HHC (71.33 ± 3.01 vs 95.79 ± 1.60, P < 0.001), 5-FU + CUR treatments (76.25 ± 3.32 vs 95.79 ± 1.60, P < 0.001) and 5-FU + HHC (68.48 ± 2.24 vs 95.79 ± 1.60, P < 0.001) markedly decreased COX-2 protein expression more than 5-FU alone. Furthermore, the AI in all treated groups, 5-FU (38.86 ± 4.73 vs 23.56 ± 2.12, P = 0.038), CUR (41.78 ± 6.92 vs 23.56 ± 2.12, P < 0.001), HHC (41.06 ± 4.81 vs 23.56 ± 2.12, P < 0.001), 5-FU + CUR (49.05 ± 6.75 vs 23.56 ± 2.12, P < 0.001) and 5-FU + HHC (53.69 ± 8.59 vs 23.56 ± 2.12, P < 0.001) significantly increased when compared to the DMH-vehicle group. However, the AI in the combination treatments, 5-FU + CUR (49.05 ± 6.75 vs 41.78 ± 6.92, P = 0.192; 49.05 ± 6.75 vs 38.86 ± 4.73, P = 0.771) and 5-FU + HHC (53.69 ± 8.59 vs 41.06 ± 4.81, P = 0.379; 53.69 ± 8.59 vs 38.86 ± 4.73, P = 0.245) did not reach significant levels as compared with each treatment alone and 5-FU monotherapy, respectively.CONCLUSION: The combined effects of HHC with 5-FU exhibit a synergistic inhibition by decreasing ACF formation mediated by down-regulation of COX-2 expression.     

Ehealth: Low FODMAP diet vs Lactobacillus rhamnosus GG in irritable bowel syndrome

AIM:To investigate the effects of a low fermentable,oligosaccharides,disaccharides,monosaccharides and polyols diet(LFD)and the probiotic Lactobacillus rhamnosus GG(LGG)in irritable bowel syndrome(IBS).METHODS:Randomised,unblinded controlled trial on the effect of 6-wk treatment with LFD,LGG or a normal Danish/Western diet(ND)in patients with IBS fulfilling Rome III diagnostic criteria,recruited betweenNovember 2009 and April 2013.Patients were required to complete on a weekly basis the IBS severity score system(IBS-SSS)and IBS quality of life(IBS-QOL)questionnaires in a specially developed IBS web selfmonitoring application.We investigated whether LFD or LGG could reduce IBS-SSS and improve QOL in IBS patients.RESULTS:One hundred twenty-three(median age 37years,range:18-74 years),90(73%)females were randomised:42 to LFD,41 to LGG and 40 to ND.A significant reduction in mean±SD of IBS-SSS from baseline to week 6 between LFD vs LGG vs ND was revealed:133±122 vs 68±107,133±122 vs 34±95,P<0.01.Adjusted changes of IBS-SSS for baseline covariates showed statistically significant reduction of IBS-SSS in LFD group compared to ND(IBS-SSS score75;95%CI:24-126,P<0.01),but not in LGG compared to ND(IBS-SSS score 32;95%CI:18-80,P=0.20).IBS-QOL was not altered significantly in any of the three groups:mean±SD in LFD 8±18 vs LGG 7±17,LFD 8±18 vs ND 0.1±15,P=0.13.CONCLUSION:LFD is efficacious for patients with IBS.