TROVE2 regulated invasion and migration of hepatocellular carcinoma cells via heparanase

Background:The role of TROVE domain family member 2(TROVE2)has been well-demonstrated in autoimmune diseases;however,its involvement in liver cancer remains unclear.Therefore,this study aimed to explore the biological function and clinical significance of TROVE2 in hepatocellular carcinoma(HCC).Methods:The expression level of TROVE2 was analyzed in HCC and paired adjacent tissue samples using real-time reverse transcription-quantitative polymerase chain reaction.The impact of TROVE2 on migration and invasion in HCC cells was analyzed through Transwell assays and Western blotting.High-throughput transcriptome sequencing and bioinformatics analyses were performed to identify downstream target genes.Back-complementation experiments were employed to verify the influence of downstream proteins on TROVE2-induced invasion and migration of HCC cells.Results:TROVE2 exhibited significant overexpression in liver cancer tissue,correlating with shorter overall survival.Overexpression of TROVE2 facilitated the invasion,metastasis,and epithelial-mesenchymal transition(EMT)process of HCC cells,whereas TROVE2 knockdown restrained migration,invasion,and EMT in these cells.Transcriptome sequencing and bioinformatics analysis identified heparanase(HPSE)as a downstreamtarget protein of TROVE2.Subsequent back-complementation experiments provided evidence that HPSE overexpression promoted TROVE2-mediated prometastasis effects.Moreover,the study revealed that TROVE2 was capable of regulating the EMT pathway through GSK-3βphosphorylation.Conclusions:TROVE2 facilitated the invasion,migration,and EMT process ofHCC cells through phosphorylation of the HPSE/GSK-3βaxis,indicating its significance as an important protein in tumor progression.

Expression levels of ROS and Atg proteins in the vitreous in rhegmatogenous retinal detachment

AIM:To detect the concentrations of reactive oxygen species(ROS),transient receptor potential mucin-1(TRPML1),and autophagy-related(Atg)proteins(LC3-Ⅰ,LC3-Ⅱ,and Beclin1)in vitreous humor of patients with simple rhegmatogenous retinal detachment(RRD).METHODS:RRD patients enrolled as the RRD group,and patients with idiopathic macular hole(IMH)and idiopathic macular epiretinal membrane(IMEM)were enrolled as control group.The levels of ROS,TRPML1,LC3-Ⅰ,LC3-Ⅱ,and Beclin1 in vitreous humor of patients in the RRD and control groups were detected by enzyme-linked immunosorbent assay(ELISA).RESULTS:The RRD group included 28 eyes 28 patients and had a higher concentration of ROS in vitreous humor(631.86±18.05 vs 436.34±108.22 IU/m L,P<0.05).The ROS level in patients with a wide retinal detachment(RD)extent(RD range≥1/2)was higher than that with a narrow RD extent(RD range<1/2,P<0.05).ROS concentration was negatively correlated with RD time(r=-0.46,P=0.01).The expression levels of LC3-Ⅰand Beclin1 significantly decreased in RRD(P<0.05),but there were no correlations with the RD time,RD extent,or macular involvement.CONCLUSION:In eyes with RRD,the concentration of ROS in vitreous humor increases and the expression levels of Atg proteins decrease,reflecting possibly that autophagy is inhibited.

低温停搏液灌注压力对猪冠状动脉内皮及平滑肌功能的影响

目的 探讨心脏停搏液灌注压力高低对猪冠状动脉血管平滑肌功能及内皮依赖性舒张反应的影响。方法 用改良的St．Thomas’hospital液分别在40、60、80mmHg的灌注压下灌注猪冠状动脉后，取2cm长的猪冠状动脉前降支（left anterior descending branch，LAD）远端血管，外径1．0—1．5mm，截成1mm长的血管环。将所取血管环直接置入器官浴槽内测定内皮及平滑肌功能。通过蓄积方式加入浓度增递的P物质测定血管内皮依赖性舒张（endothelium dependent relaxation，EDR）功能。结果 在本实验灌注压力下，冠状动脉平滑肌的舒缩功能均不受影响（P＞0．05）。与新鲜对照组血管[（98．0±1．2）％]相比，40mmHg和60mmHg组冠状动脉的最大EDR值无显著下降[分别为（96．7±1．5）％、（96．6±1．9）％、（P＞0．05）]，80mmHg组血管的最大EDR值则显著降低[（90．8±2．2）％，（P＜0．01）]。在血管内皮对P物质的敏感性（用50％最大舒张时的浓度值表示，pEC50）方面，与新鲜对照组对比，40mmHg组血管环pEC50值无显著降低（P＞0．05），60mmHg组和80mmHg组血管环的pEC50值则显著降低（P＜0．05）、（P＜0．01）。结论 用改良的St．Thomas’hospital溶液灌注猪心时，60mmHg的压力即可以造成冠状动脉内皮功能的损害，而且灌注压力越高，损伤越重。

Heat shock protein 70 antisense oligonucleotide inhibits cell growth and induces apoptosis in human gastric cancer cell line SGC-7901

AIM: Heat shock protein (HSP)70 is over-expressed in human gastric cancer and plays an important role in the progression of this cancer. We investigated the effects of antisense HSP70 oligomer on human gastric cancer cell line SGC-7901, and its potential role in gene therapy for this cancer.METHODS: Human gastric cancer cell line SGC-7901 was treated in vitro with various concentrations of antisense HSP70 oligonucleotides at different intervals. Growth inhibition was determined as percentage by trypan blue dye exclusion test. Extracted DNA was electrophoresed on agarose gel, and distribution of cell cycle and kinetics of apoptosis induction were analyzed by propidium iodide DNA incorporation using flow cytometry, which was also used to detect the effects of antisense oligomer pretreatment on the subsequent apoptosis induced by heat shock in SGC-7901 cells. Proteins were extracted for simultaneous measurement of HSP70 expression level by SDS-PAGE Western blotting.RESULTS: The number of viable cells decreased in a doseand time-dependent manner, and ladder-like patterns of DNA fragments were observed in SGC-7901 cells treated with antisense HSP70 oligomers at a concentration of 10 μmol/L for 48 h or 8 μmol/L for 72 h, which were consistent with inter-nucleosomal DNA fragmentation. Flow cytometric analysis showed a dose- and time-dependent increase in apoptotic rate by HSP70 antisense oligomers. This response was accompanied with a decrease in the percentage of cells in the G1 and S phases of the cell cycle, suggesting inhibition of cell proliferation. In addition, flow cytometry also showed that pretreatment of SGC-7901 cells with HSP70 antisense oligomers enhanced the subsequent apoptosis induced by heat shock treatment. Western blotting demonstrated that HSP70 antisense oligomers inhibited HSP70 expression, which preceded apoptosis, and HSP70 was undetectable at the concentration of 10 μmol/L for 48 h or 8 μmol/L for 72 h.CONCLUSION: Antisense HSP70 oligomers can abrogate HSP70 expression in SGC-7901 cells, which may in turn induce apoptosis and inhibit cell proliferation, conversely suggesting that HSP70 is required for the proliferation and survival of human gastric cancer cells under normal conditions.

Risk factors and gene polymorphisms of inflammatory bowel disease in population of Zhejiang,China

AIM:To identify the risk factors and three single nucleotide polymorphisms(SNPs) of NOD2/CARD15 gene in inflammatory bowel disease(IBD) of the population in Zhejiang,China.METHODS:A case-control study was conducted using recall questionnaire to collect data on demographic,socioeconomic,lifestyle characteristics and dietary behaviors from 136 determined IBD patients and 136 paired healthy controls.COX regression method was used to screen the statistically significant risk factors for IBD.The polymorphisms of NOD2/CARD15 gene Arg702Trp,Gly908Arg and Leu1007fsinsC were genotyped and further compared between 60 patients with IBD and 60 healthy controls by polymerase chain reaction and restriction fragment length polymorphism.RESULTS:IBD occurred primarily in young and middle-aged people.The mean age for IBD patients was 42.6 years.The ratio of males to females was 1.23:1.COX regression indicated a higher statistical significance in milk,fried food and stress compared with the other postulated risk factors for IBD.None of the patients with IBD and healthy controls had heterozygous or homozygous SNPs variants.CONCLUSION:Milk,fried food and stress are associated with increased risk of IBD.The common variants in NOD2/CARD15 gene are not associated with IBD in China's Zhejiang population.

Epidemiological and histopathological study of relevance of Guizhou Maotai liquor and liver diseases

AIM：To explore the relevance of Maotai liquor and liver diseases.METHODS:Epidemiological study was conducted on groups of subjects,each consisting of 3 subjects from the Maotai liquor roup consisting of 99 individuals and one from the non-alcoholic control group consisting of 33 individuals Liver biopsy was performed on 23 volunteere from Guizhou Maotai Distillery who had a constant and long history of drinking Maotai liquor.Experimental histopathological study was conducted as follows:Sixty male Wistar rate were divided into 3 groups randomly and fed with Maotai liquor,ordinary white wine,and physiological saline respectively for a period of 8and 12 weeks,The rats were sacrificed in batches,then serum ALT,AST,TBil,and AKP were measured.Rat livers were harvested to Measure the liver indexes GSH,and MDA.Histopathological examinations were also performed.Another eighty mice were randomly divided into 4 groups and fed with Maotai(at different dosages of 10ml.kg^-1 and 20ml.kg^-1),ethanol,and physiological saline.The animals were sacrificed after 4 weeks and serum ALT was determined ,Then the livers were harvested and liver indexes and MDA were measured.RESULTS:The incidence rate of hepatic symptoms splenomegaly,liver function impairment,reversal of Albumin/Globulin and increased diameter of portal verins in the Maotai liquor group were 1.0%(1/99),1.0%(1/99),1.0%(1/99),1.0%(1/99),9(9/99)and 9(0/99),0(0/99),0(0/99),0(0/99),0(0/99),respectively,There was no significant difference between the Maotai group[ and the non-alcoholic control group(P>0\05),Various degree of fatty infiltration of hepatocytes was found in the 23 volunteers receiving liver biopsy.but there was no obvious hepatic fibrosis or cirrhosis.A comparison was made between the Maotai liquor group and the ordinary white wine group,It was found that hepatic MAD in rats and mice,were 0.33±0.10and 0.49±0.23respectively in Maotai group and 0.61±0.22and 0.66±0.32inthe ordinary white wine group;MDA had an obvious decrease in the Maotai liquor group(P<0.05)hepatic GSH were 0.12mg.g^-1±0.06mg.g^-1 in rats of the Maotai liquor group and (0.08±0.02)mg.g^-1 in white wine group,it was obviously increased in the Maotai liquor group(P<0.05),Atfer the 20 rats had been fed with ordinary white wine for 8 Weeks consecutively,disarranged hepatocyte cords,fatty infiltration of hepatocytes,and fibrous septa of varying widths due to hepatic connective tissues proliferation were observed;after 12 weeks,the fibrous tissue proliferation continued and early cirrhosis appeared.COmpared with the ordinary whtite wine group fatty infiltration was observed in the 8-week and 12-week groups,but no necrosis or filbrosis or cirrhosis was found in the Maotai liquor group(P<0.05）。CONCLUSION:Maotai liquor may cause fatty liver but not hepatic fibrosis or cirrhosis,and it can strengthen lipid peroxidation in the liver。

Heme oxygenase-1 prevents liver fibrosis in rats by regulating the expression of PPAR_γ and NF-_κB

AIM:To investigate the effects of heme oxygenase(HO)-1 on liver fibrosis and the expression of peroxisome proliferator-activated receptor gamma(PPARγ) and nuclear factor-kappa B(NF-κB) in rats.METHODS:Sixty Wistar rats were used to construct liver fibrosis models and were randomly divided into 5 groups:group A(normal,untreated),group B(model for 4 wk,untreated),group C(model for 6 wk,untreated),group D [model for 6 wk,treated with zinc protoporphyrin Ⅸ(ZnPP-Ⅸ) from week 4 to week 6],group E(model for 6 wk,treated with hemin from week 4 to week 6).Next,liver injury was assessed by measuring serum alanine aminotransferase(ALT),aspartate aminotransferase(AST) and albumin levels.The degree of hepatic fibrosis was evaluated by measuring serum hyaluronate acid(HA),type Ⅳ collagen(Ⅳ-C) and by histological examination.Hydroxyproline(Hyp) content in the liver homogenate was determined.The expres-sion levels of alpha-smooth muscle actin(α-SMA) in liver tissue were measured by real-time quantitative polymerase chain reaction(RT-PCR).The expression levels of PPARγ and NF-κB were determined by RT-PCR and Western blotting.RESULTS:The expression of HO-1 increased with the development of fibrosis.Induction of HO-1 by hemin significantly attenuated the severity of liver injury and the levels of liver fibrosis as compared with inhibition of HO-1 by ZnPP-Ⅸ.The concentrations of serum ALT,AST,HA and Ⅳ-C in group E decreased compared with group C and group D(P < 0.01).Amount of Hyp and α-SMA in the liver tissues in group E decreased compared with group C(0.62 ± 0.14 vs 0.84 ± 0.07,1.42 ± 0.17 vs 1.84 ± 0.17,respectively,P < 0.01) and group D(0.62 ± 0.14 vs 1.11 ± 0.16,1.42 ± 0.17 vs 2.56 ± 0.37,respectively,P < 0.01).The expression of PPARγ at levels of transcription and translation decreased with the development of fibrosis especially in group D;and it increased in group E compared with groups C and D(0.88 ± 0.15 vs 0.56 ± 0.19,0.88 ± 0.15 vs 0.41 ± 0.11,respectively,P < 0.01).The expression of NF-κB increased with the development of fibrosis especially in group D;and it decreased in group E compared with groups C and D(1.43 ± 0.31 vs 1.89 ± 0.29,1.43 ± 0.31 vs 2.53 ± 0.54,respectively,P < 0.01).CONCLUSION:Our data demonstrate a potential mechanism that HO-1 can prevent liver fibrosis by enhancing the expression of PPARγ and decreasing the expression of NF-κB in liver tissues.

Expression of Livin and vascular endothelial growth factor in different clinical stages of human esophageal carcinoma

AIM: To investigate the role of Livin and vascular endothelial growth factor (VEGF) in human esophageal carcinoma, and analyze its relationship to clinical stages. METHODS: Expression of Livin in fresh esophageal cancer tissues was detected by immunohistochemistry (IHC), Western blotting and reverse transcriptase-polymerase chain reaction (RT-PCR), and VEGF by Western blotting and RT-PCR. All statistical analyses were performed by SPSS version 11.0. RESULTS: Livin positivity was also significantly correlated with tumor stages, increasing with tumor progression. Expression of Livin and VEGF increased with the process of esophageal carcinoma. In the fourth clinical stage, expression of Livin and VEGF was the most significant. Expression of Livin was positively correlated with VEGF. CONCLUSION: Over-expression of Livin and VEGF contributes to the pathogenesis of esophageal carcinoma.

Metastasis-associated protein 1 induces VEGF-C and facilitates lymphangiogenesis in colorectal cancer

AIM:To study the correlation between high metastasisassociated protein 1(MTA1)expression and lymphangiogenesis in colorectal cancer(CRC)and its role in production of vascular endothelial growth factor-C(VEGF-C). METHODS:Impact of high MTA1 and VEGF-C expression levels on disease progression and lymphovasculardensity(LVD,D2-40-immunolabeled)in 81 cases of human CRC was evaluated by immunohistochemistry. VEGF-C mRNA and protein expressions in human LoVo and HCT116 cell lines were detected by real-time polymerase chain reaction and Western blotting,respectively,with a stable expression vector or siRNA. RESULTS:The elevated MTA1 and VEGF-C expression levels were correlated with lymph node metastasis and Dukes stages(P<0.05).Additionally,high MTA1 expression level was correlated with a large tumor size(P< 0.05).A significant correlation was found between MTA1 and VEGF-C protein expressions in tumor cells(r=0.371, P<0.05).Similar to the VEGF-C expression level,high MTA1 expression level was correlated with high LVD in CRC(P<0.05).Furthermore,over-expression of MTA1 significantly enhanced the VEGF-C mRNA and protein expression levels,whereas siRNAs-knocked down MTA1 decreased the VEGF-C expression level. CONCLUSION:MTA1,as a regulator of tumor-associated lymphangiogenesis,promotes lymphangiogenesis in CRC by mediating the VEGF-C expression.

Effect of Maotai liquor in inducing metallothioneins and on hepatic stellate cells

AIM: To explore the possible mechanism why drinkingMaotai liquor dose not cause hepatic fibrosis.METHODS: After being fed with Maotai for 56 daysconsecutively, the male SD rats were decollated fordetecting the biological indexes, and the livers wereharvested to examine the liver indexes and the level ofhepatic metallothioneins (MT). Hepatic stellate cells (HSC)proliferation and collagen generation were also observed.RESULTS: Hepatic MT contents were 216.0 ng@g1 + 10.8 ng@g1 in the rats of Maotai group and 10.0 ng@g-1 ± 2.8 ng@g1 inthe normal control group, which was increased obviously inMaotain group ( P ＜ 0.05). In the rats with grade CCL2poisoning induced by Maotai, hepatic MT content was 304.8ng@ g1 + 12.1 ng@ g-1 whereas in the controls with grade CCL4poisoning, it was 126.4 ng@g-1 +4.8 ng@g-1(P＜0.05).MDAwas 102.0 nmol@ g-1 + 3.4 nmol@ g-1 in Maotai group and 150.8nmol@ g-1 + 6.7 nmol@ g1 in the control group ( P ＜ 0.05).When both of the groups were suffering from grade CCL4poisoning, hepatic MT contents was negatively correlatedwithMDA (r=-0.8023, n=20, P＜0.01). The 570 nmAvalues of each tube with HSC regeneration at concentrationsof 0, 10, 50, 100, and 200g@L-1 of Maotai were 0.818, 0.742,0.736, 0.72, 0.682, and 0.604, respectively. From theconcentration of 10 g@ L1, Maotai began to show obviousinhibitory effects against HSC, and the inhibition wasconcentration-dependent (P＜0.05, P＜ 0.01). Type Icollagen contents in HSC were 61.4, 59.9, 50.1, 49.2, 48.7,34.4μg@g1 at concentrations of 0, 10, 50, 100, and 200g@ L-1of Maotai. At the concentration of 100-200 g@L-1, Maotai hadobvious inhibitory effect against the secretion of type Icollagen (P ＜ 0. 05 ). Gene expression analysis wasconducted on cells with Maotai concentrations of 0, 50, 100g@L-1 respectively and the ash values of β-actin geneexpression were 0. 88, 0. 74, and 0. 59, respectively,suggesting that at the concentration of 100 g@ L-1 , Maotaicould obviously inhibit gene expression of type I procollagen(P＜ 0.05), but the effect was not obvious at theconcentration of 50 g@L-1(P＞0.05). At the concentration of10 g@L-1, HSC growth in vitro inhibition rates were 16.4+2.3 inMaotai group and-8.4+ 2.3 in the control group (P＜0.05).CONCLUSION: Maotai Iiluor can increase metatlothioneins inthe liver and inhibit the activation of HSC and the synthesisof collagen in many aspects, which might be the mechanismthat Maotai liquor interferes in the hepatic fibrosis.

Drug-induced liver injury in hospitalized patients with notably elevated alanine aminotransferase

AIM:To identify the proportion,causes and the nature of drug-induced liver injury(DILI) in patients with notably elevated alanine aminotransferase(ALT).METHODS:All the inpatients with ALT levels above 10 times upper limit of normal range(ULN) were retrospectively identified from a computerized clinical laboratory database at our hospital covering a 12-mo period.Relevant clinical information was obtained from medical records.Alternative causes of ALT elevations were examined for each patient,including biliary abnormality,viral hepatitis,hemodynamic injury,malignancy,DILI or undetermined and other causes.All suspected DILI cases were causality assessed using the Council for International Organizations of Medical Sciences scale,and only the cases classified as highly probable,probable,or possible were diagnosed as DILI.Comments related to the diagnosis of DILI in the medical record and in the discharge letter for each case were also examined to evaluate DILI detection by the treating doctors.RESULTS:A total of 129 cases with ALT > 10 ULN were identified.Hemodynamic injury(n = 46,35.7%),DILI(n = 25,19.4%) and malignancy(n = 21,16.3%) were the top three causes of liver injury.Peak ALT values were lower in DILI patients than in patients with hemodynamic injury(14.5 ± 5.6 ULN vs 32.5 ± 30.7 ULN,P = 0.001).Among DILI patients,one(4%) case was classified as definite,19(76%) cases were classified as probable and 5(20%) as possible according to the CIOMS scale.A hepatocellular pattern was observed in 23(92%) cases and mixed in 2(8%).The extent of severity of liver injury was mild in 21(84%) patients and moderate in 4(16%).Before discharge,10(40%) patients were recovered and the other 15(60%) were improved.The improved patients tended to have a higher peak ALT(808 ± 348 U/L vs 623 ± 118 U/L,P = 0.016) and shorter treatment duration before discharge(8 ± 6 d vs 28 ± 12 d,P = 0.008) compared with the recovered patients.Twenty-two drugs and 6 herbs were found associated with DILI.Antibacterials were the most common agents causing DILI in 8(32%) cases,followed by glucocorticoids in 6(24%) cases.Twenty-four(96%) cases received treatment of DILI with at least one adjunctive drug.Agents for treatment of DILI included anti-inflammatory drugs(e.g.,glycyrrhizinate),antioxidants(e.g.,glutathione,ademetionine 1,4-butanedisulfonate and tiopronin),polyene phosphatidyl choline and herbal extracts(e.g.,protoporphyrin disodium and silymarin).Diagnosis of DILI was not mentioned in the discharge letter in 60% of the cases.Relative to prevalent cases and cases from wards of internal medicine,incident cases and cases from surgical wards had a higher risk of missed diagnosis in discharge letter [odds ratio(OR) 32.7,95%CI(2.8-374.1),CONCLUSION:DILI is mostly caused by use of antibacterials and glucocorticoids,and constitutes about one fifth of hospitalized patients with ALT > 10 ULN.DILI is underdiagnosed frequently.

B7-H1 expression is associated with expansion of regulatory T cells in colorectal carcinoma

AIM:To investigate the expression of B7-H1 in human colorectal carcinoma (CRC) to define its regulating effects on T cells in tumor microenvironment.METHODS:One hundred and two paraffin blocks and 33 fresh samples of CRC tissues were subject to this study.Immunohistochemistry was performed for B7-H1 and CD3 staining in CRC tissues.Ficoll-Hypaque density gradient centrifugation was used to isolate peripheral blood mononuclear cells of fresh CRC tissues;flow cytometry and immunofluorescence staining were used for detection of regulatory T cells.Data was analyzed with statistical software.RESULTS:Costimulatory molecule B7-H1 was found strongly expressed in CRC tissues,localized in tumor cell membrane and cytoplasm,while weak or none expression of B7-H1 was detected in pared normal colorectal tissues.Meanwhile,CD3 positive T cells were found congregated in CRC tumor nest and stroma.Statistic analysis showed that B7-H1 expression level was negatively correlated to the total T cell density in tumor nest (P<0.0001) and tumor stroma (P=0.0200) of 102 cases of CRC tissues.Among the total T cells,a variable amount of regulatory T cells with a clear Foxp3+ (forkhead box P3) staining could be detected in CRC tissues and patients' blood.Interestingly,in the 33 samples (15 cases of B7-H1 high CRC tissues and 18 cases of B7H1 low CRC tissues) of freshly isolated mononuclear cells from CRC tissues,the percentages of CD4+ Foxp3+ and CD8+ Foxp3+ regulatory T cells were found remarkably higher in B7-H1 high CRC tissues than in B7-H1 low CRC tissues (P=0.0024,P=0.0182),indicating that B7-H1 expression was involved in proliferation of regulatory T cell.No significant difference was found in CRC peripheral blood (P=0.0863,P=0.0678).PD-1 is the specific ligand for B7-H1 pathway transferring inhibitory signal to T cell,which is expressed by activated T cell.Our further analysis of PD-1 expression on T cells in CRC tissues showed that conventional T cells (CD4+ Foxp3/CD8+ Foxp3),which was thought to contribute to the anti-tumor immune response,highly expressed PD-1;while regulatory T cells (CD4+ Foxp3+/CD8+ Foxp3) almost failed to express PD-1.The average percentage of PD-1 expression on regulatory T cells was significantly higher than the percentage of PD-1 on conventional T cells (CD4+ Foxp3 T cell,P<0.0001;CD8+ Foxp3 T cell,P<0.0001).The diverse expression of PD-1 might lead to different fate of T cell subsets in B7-H1 over-expression CRC tumor microenvironment.CONCLUSION:B7-H1 expression in tumor cells can in hibit the conventional T cell proliferation in tumor micro environment through the PD-1 expression on conventiona T cells.

Vitamin D receptor gene polymorphisms and colorectal cancer risk:A systematic meta-analysis

AIM:To investigate the relationship between polymorphisms present in the vitamin D receptor(VDR) gene and colorectal cancer risk,a systematic meta-analysis of population-based studies was performed.METHODS:A total of 38 relevant reports published between January 1990 and August 2010 were identified,of which only 23 qualified for this meta-analysis based on our selection criteria.Five polymorphic variants of the VDR gene,including Cdx-2(intron 1e) and FokI(exon 2) present in the 5' region of the gene,and BsmI(intron 8),ApaI(intron 8),and TaqI(exon 9) sites present in the 3' untranslated region(UTR),were evaluated for possible associations with colorectalcancer risk.Review manager 4.2 was used to perform statistical analyses.RESULTS:In the meta-analysis performed,only the BsmI polymorphism was found to be associated with colorectal cancer risk.In particular,the BsmI B genotype was found to be related to an overall decrease in the risk for colorectal cancer [BB vs bb:odds ratio(OR) = 0.87,95% CI:0.80-0.94,P = 3 × 10-4;BB vs Bb + bb:OR = 0.90,95% CI:0.84-0.97,P = 5 × 10-4].Moreover,in subgroup analyses,the BsmI B genotype was significantly associated with colon cancer,and not rectal cancer.An absence of between-study heterogeneity was also observed.CONCLUSION:A meta-analysis of 23 published studies identified the BsmI polymorphism of the VDR gene to be associated with an increased risk of colon cancer.

Thermotherapy enhances oxaliplatin-induced cytotoxicity in human colon carcinoma cells

AIM:To observe the synergistic effects of hyperthermia in oxaliplatin-induced cytotoxicity in human colon adenocarcinoma Lovo cells.METHODS:The human colon adenocarcinoma cell line Lovo was obtained from Sun Yat-Sen University.Cells were sealed with parafilm and placed in a circulating water bath,and was maintained within 0.01 ℃ of the desired temperature (37 ℃,39 ℃,41 ℃,43 ℃ and 45 ℃).Thermal therapy was given alone to the negative control group while oxaliplatin was administered to the treatment group at doses of 12.5 μg/mL and 50 μg/mL.Identification of morphological changes,3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyltetrazolium bromide assay,flow cytometry and Western blotting were used to investigate the effect of thermochemotherapy on human colon adenocarcinoma Lovo cells,including changes in the signal pathway related to apoptosis.RESULTS:A temperature-dependent inhibition of cell growth was observed after oxaliplatin exposure,while a synergistic interaction was detected preferentially with sequential combination.Thermochemotherapy changed the morphology of Lovo cells,increased the inhibition rate of the Lovo cells (P < 0.05) and enhanced cellular population in the G 0 /G 1 phase (16.7% ± 4.8 % in phase S plus 3.7% ± 2.4 % in phase G 2 /M,P < 0.05).Thermochemotherapy increased apoptosis through upregulating p53,Bax and downregulating Bcl-2.Protein levels were elevated in p53,Bax/Bcl-2 in thermochemotherapy group as compared with the control group (P < 0.05).CONCLUSION:Thermochemotherapy may play an important role in apoptosis via the activation of p53,Bax and the repression of Bcl-2 in Lovo cells.

Inhibitory effect of Polo-like kinase 1 depletion on mitosis and apoptosis of gastric cancer cells

瞄准：像马球的激酶 1 (PLK1 ) serine/threonine 激酶在胃的癌症房间在有丝分裂的多重阶段起一个重要作用。在胃的癌症房间的有丝分裂和 apoptosis 上调查 PLK1 弄空的效果。方法：PLK1 表示被小 RNA 干扰(siRNA ) 堵住。PLK1， cdc2， cyclin B 和 caspase 的表示层次 3 被西方的弄污检测。然后， PLK1 击倒的房间的 apoptosis 的 PLK1 弄空， cdc2 活动，房间增长，房间周期阶段分发，有丝分裂的锭子结构，和率被观察。结果：PLK1 基因击倒与增加的 cyclin B 表示被联系，增加的 cdc2 活动(然而并非与表示层次) ，在 G2/M，不适当的有丝分裂的锭子形成，推迟的染色体分离和推迟或逮捕的胞质分裂的胃的癌症房间的累积。而且，在胃的癌症房间的 PLK1 弄空与减少的增长， 3 铺平的稀释 pro-caspase 和增加的 apoptosis 被联系。结论：PLK1 表示的阻塞可以在胃的癌症房间导致减少的有丝分裂或甚至 apoptosis，显示 PLK1 可以是为胃的癌症的一个珍贵治疗学的目标。

Effect of smoking on semen quality of infertile men in Shandong, China

Aim: To study the effect of smoking on the semen quality in infertile men in Shandong Province, China. Methods:Adult non-drinker males attending the infertility clinic, including 110 non-smokers and 191 smokers, were recruited forthe study. Sixty-one fertile, non-smoker and non-drinker males, who had one or more children, served as the controls.The smokers were divided into subgroups according to the amount and duration of smoking. Semen parameters (semenvolume and sperm density, viability, motility, and morphology) were examined and seminal plasma contents of Zn,Cu and superoxide dismutase (SOD) determined. Results: The semen volume and acidity, and the sperm density,viability and forward progression, as well as the seminal plasma contents of Zn, Cu and SOD were much lower in themedium, heavy and long-term smokers than in the non-smokers ( P < 0.01). The sperm density, viability and forwardprogression, and the seminal plasma Zn, Cu and SOD levels were negatively correlated with the amount and duration ofcigarette smoking (P < 0.01). Conclusion: Medium, heavy and long-term smoking adversely affected the semenquality in a population of men visiting the infertility clinic in Shandong, China.

Tuberculosis versus non-Hodgkin's lymphomas involving small bowel mesentery:Evaluation with contrast-enhanced computed tomography

AIM: To evaluate the specific computed tomography (CT) imaging criteria for differentiating tuberculosis involving the small bowel mesenteric lymph nodes from lymphomas. METHODS: We retrospectively reviewed the anatomic distribution,CT enhancement patterns of lymphoma in 18 patients with mesenteric tuberculosis and 22 with untreated non-Hodgkin’s lymphomas (NHL) involving small bowel mesentery (SBM). Of the 18 patients with tuberculosis,9 had purely mesenteric tuberculous lymphadenopathy (TL),and 9 had mesenteric TL accompanied with tuberculous mesenteritis (TLM). RESULTS: CT showed that tuberculosis and NHL mainly affected lymph nodes in the body and root of SBM. Homogeneously enhanced lymph nodes in the body and root of SBM were found more often in the NHL (P < 0.05). Homogeneously mixed peripheral enhanced lymph nodes in the body of SBM were found more often in mesenteric TL and TLM (P < 0.05). Peripheral enhanced lymph nodes in the root of SBM were found more often in mesenteric TL and TLM (P < 0.01). "Sandwich sign" in the root of SBM was observed more often in NHL (P < 0.05). CONCLUSION: Anatomic lymph node distribution,sandwich sign and specific enhancement patterns of lymphadenopathy in SBM on CT images can be used in differentiating between tuberculosis and untreated NHL involving SBM.

Protective effects of non-mitogenic human acidic fibroblast growth factor on hydrogen peroxide-induced damage to cardiomyocytes in vitro

AIM: To study the protective effect of non-mitogenic human acidic fibroblast growth factor (FGF) on cardiac oxidative injury in vivo.METHODS: Ventricular cardiomyocytes were isolated from 1- to 3-d-old neonatal SD mice and cultured in Dulbecco's minimum essential medium supplemented with 15% fetal bovine serum under an atmosphere of 50 mL/L CO2-95% air at 37 ℃, as well as assessed by immunocytochemical assay. We constructed the cardiomyocyte injury model by exposure to a certain concentration of H2O2.Cellular viability, superoxide dismutase (SOD) activity,leakage of maleic dialdehyde and anti-apoptosis effect were included to evaluate the cardiac protective effect of non-mitogenic human acidic FGF.RESULTS: Over 50% of the cardiomyocytes beat spontaneously on the 2nd d of culture and synchronously beat after being cultured for 3 d. Forty-eight hours after plating was completed, the purity of such cultures was 95% myocytes, assessed b,y an immunocytochemical assay. Cellular viability dramatically decreased with the increasing of the concentration of H2O2. Non-mitogenic human acidic FGF showed significant resistance to the toxic effect of H2O2, significantly increased the cellular viability as well as the activity of SOD, and dramatically decreased the leakage of maleic dialdehyde as well as the cellular apoptosis rate.CONCLUSION: Hydrogen peroxide shows strong cytotoxicity to the cultured cardiac myocytes, and non-mitogenic human acidic FGF shows strong cardio-protective effect when exposed to a certain concentration of H2O2.

Sonographic features of duodenal lipomas in eight clinicopathologically diagnosed patients

AIM:To investigate the sonographic features and diagnostic value of endoscopic ultrasonography (EUS) for duodenal lipomas (DLs).METHODS:A total of eight consecutive patients with DL diagnosed pathologically were included in the study.One EUS expert reviewed the ultrasonic images for all lesions,including the original layer of the duodenal wall,the echo intensity and the echo homogeneity.The size of the lesions and the perifocal structures were also investigated.The diagnosis by EUS was compared with the histological results.RESULTS:Using routine endoscopy,only one case was correctly diagnosed as DL.Four cases were classified as submucosal tumors,and three cases were mistaken for stromal tumors.All tumors appeared as round or oval intensive hyperechoic lesions with distinct anterior borders that originated from the submucosal layer on EUS.Tumors ranged from 8 to 36 mm in size,with an average size of 16 mm.Homogeneous echogenicity was seen in all cases except one that had a tubular structure inside the tumor.Echo attenuation was observed only in the area behind the tumors in five cases,and it was observed both inside and behind the tumors in three cases in which the posterior border was obscure or invisible.Seven (87.5%) cases were correctly diagnosed as DL,and one (12.5%) was mistaken as Brunner's gland adenoma by EUS.Pathologically,all tumors originated from the submucosal layer and consisted of mature fat cells without heteromorphism.Among the fat cells,there was a small amount of thick-wall vessels infiltrating the lymphocytes,and abundant fibrous connective tissues.CONCLUSION:On EUS,DL is featured as an intensive homogeneous hyperechoic submucosal lesion with marked echo attenuation and without involvement of the mucosa.

NT4(Si)-p53(N15)-antennapedia induces cell death in a human hepatocellular carcinoma cell line

AIM:To construct the recombinant lentivirus expression plasmid,pLenti6/V5-NT4 p53(N15)-antennapedia(Ant),and study its effect on HepG2 cells.METHODS:Plasmid pLenti6/V5-NT4 p53(N15)-Ant was constructed incorporating the following functional regions,including signal peptide sequence and pro-region of neurotrophin 4,N-terminal residues 12-26 of p53 and 17 amino acid drosophila carrier protein,Ant.Hepatocellular carcinoma(HepG2)cells were used for transfection.3-[4,5-dimethyl-thiazol-2yl]-2,5 diphenyl tetrazolium bromide(MTT)assay,lactate dehydrogenase(LDH)release assay,transmission electron microscopy(TEM)and flow cytometric analysis(FCM)were employed to investigate the effects of LV-NT4(Si)-p53(N15)-Ant in vitro on HepG2 cells.In vivo experiment was also performed to investigate the inhibitory effect of LV-NT4(Si)-p53(N15)-Ant on tumor growth in nude mice.RESULTS:LV-NT4(Si)-p53(N15)-Ant significantly suppressed the growth of HepG2 cells.MTT assay showed that the growth of HepG2 cells was mucj more significantly inhibited by LV-NT4(Si)-p53(N15)-Ant than by LV-EGFP.The inhibition rate for HepG2 cell growth in the two groups was 46.9% and 94.5%,respectively,48 h after infection with LV-NT4(Si)-p53(N15)-Ant,and was 33.9% and 95.8%,respectively,72 h after infection with LV-NT4(Si)-p53(N15)-Ant(P < 0.01).Light microscopy and TEM showed morphological changes in HepG2 cells infected with LV-NT4(Si)-p53(N15)-Ant,but no signif icant changes in HepG2 cells infected with LV-EGFP.Changes were observed in ultra-structure of HepG2 cells infected with LV-NT4(Si)-p53(N15)-Ant,with degraded membranes,resulting in necrosis.LDH release from HepG2 cells was analyzed at 24,48,72 and 96 h after infection with LV-NT4(Si)-p53(N15)-Ant and LV-EGFP,which showed that LDH release was signif icantly higher in LV-NT4(Si)-p53(N15)-Ant treatment group(682 IU/L)than in control group(45 IU/L,P < 0.01).The longer the time was after infection,the bigger the difference was in LDH release.FCM analysis showed that LV-NT4(Si)-p53(N15)-Ant could induce two different kinds of cell death:necrosis and apoptosis,with apoptosis being the minor type and necrosis being the main type,suggesting that LV-NT4(Si)-p53(N15)-Ant exerts its anticancer effect on HepG2 cells by inducing necrosis.The in vivo study showed that LV-NT4(Si)-p53(N15)-Ant significantly inhibited tumor growth with an inhibition rate of 66.14% in terms of tumor size and weight.