Molecular mechanisms of ＂off-on switch＂ of activities of human IDH1 by tumor-associated mutation R132H

Human cytosolic NADP-1DH （IDH1） has recently been found to be involved in tumorigenesis. Notably, the tumorderived IDH1 mutations identified so far mainly occur at Arg132, and mutation R132H is the most prevalent one. This mutation impairs the oxidative IDH activity of the enzyme, but renders a new reduction function of converting a-ketoglutarate （aKG） to 2-hydroxyglutarate. Here, we report the structures of the R132H mutant IDH1 with and without isocitrate OCT） bound. The structural data together with mutagenesis and biochemical data reveal a previ- ously undefined initial ICT-binding state and demonstrate that IDH activity requires a conformational change to a closed pre-transition state. Arg132 plays multiple functional roles in the catalytic reaction; in particular, the R132H mutation hinders the conformational changes from the initial ICT-binding state to the pre-transition state, leading to the impairment of the IDH activity. Our results describe for the first time that there is an intermediate conformation that corresponds to an initial ICT-binding state and that the R132H mutation can trap the enzyme in this conforma- tion, therefore shedding fight on the molecular mechanism of the ＂off switch＂ of the potentially tumor-suppressive IDH activity. Furthermore, we proved the necessity of Tyr139 for the gained aKG reduction activity and propose that Tyr139 may play a vital role by compensating the increased negative charge on the C2 atom of aKG during the trans- fer of a hydride anion from NADPH to aKG, which provides new insights into the mechanism of the ＂on switch＂ of the hypothetically oncogenic reduction activity of IDH1 by this mutation.

Molecular evaluation of glutathione S transferase family genes in patients with sporadic colorectal cancer

AIM To evaluate the association between polymorphismsin glutathione S transferases(GSTs) and the risk of sporadic colorectal cancer(SCRC), tumor progression and the survival of patients.METHODS A case-control study of 970 individuals from the Brazilian population was conducted(232 individuals from the case group with colorectal cancer and 738 individuals from the control group without a history of cancer). PCR multiplex and PCR-RFLP techniques were used to genotype the GST polymorphisms. The tumors were categorized according to the TNM classification: tumor extension(T), affected lymph nodes(N), and presence of metastasis(M). Logistic regression, multiple logistic regression and survival analysis were used to analyze the data. The results are presented in terms of odds ratio(OR) and 95% confidence interval(CI). The level of significance was set at 5%(P ≤ 0.05).RESULTS Age equal to or over 62 years(OR = 8.79; 95%CI: 5.90-13.09, P < 0.01) and female gender(OR = 2.91; 95%CI: 1.74-4.37; P < 0.01) were associated with increased risk of SCRC. Analysis of the polymorphisms revealed an association between the GSTM1 polymorphisms and a risk of SCRC(OR = 1.45; 95%CI: 1.06-2.00; P = 0.02), as well as between GSTT1 and a reduced risk of the disease(OR = 0.65; 95%CI: 0.43-0.98; P = 0.04). An interaction between the presence of the wild-type allele of GSTP1 Ile105 Val polymorphism and tobacco consumption on risk of SCRC(OR = 2.33; 95%CI: 1.34-4.05; P = 0.05) was observed. There was an association between the GSTM1 null genotype and the presence of advanced tumors(OR = 2.33; 95%CI: 1.23-4.41; P = 0.009), as well as increased risk of SCRC in the presence of a combination of GSTT1 non-null/GSTM1 null genotypes(OR = 1.50; 95%CI: 1.03-2.19; P = 0.03) and GSTT1 non-null/GSTM1 null/GSTP1 Val~*(OR = 1.85; 95%CI: 1.01-3.36, P = 0.04). Combined GSTT1 non-null/GSTM1 null genotypes(OR = 2.40; 95%CI: 1.19-4.85; P = 0.01) and GSTT1 non-null/GSTM1 null/GSTP1 Val~*(OR = 2.92; 95%CI: 1.05-8.12; P = 0.04) were associated with tumor progression. Polymorphisms were not associated with the survival of patients with SCRC.CONCLUSION Females aged 62 years or older are more susceptible to SCRC. Polymorphisms of GSTT1 and GSTM1 null genotypes modulated the susceptibility to SCRC in the population studied.

Elicitation-Based Modulation of Shelf Life in Fruits: Physiological and Molecular Insights

The process of ripening involves physiological and biochemical events that become a concern during postharvest storage.We have documented different approaches for the preservation and maintenance of fruit quality during the postharvest period that are biocompatible and fully safe for consumption.Chemical residues that sustain sensory characteristics,such as color,flavor,aroma,and texture,are considered.In fruit ripening,both physical and chemical elicitors are described that regulate ethylene biosynthesis or its signaling for gene expression.The key regulatory enzymes,such as ACC synthase and ACC oxidase,for ethylene biosynthesis,are important for both climacteric and non-climacteric fruits.Anti-oxidizing genes that retain sensory characteristics are concerns in this respect.Chemical elicitors,including chitosan,polyamine,phenolics,lipopolysaccharide,silver derivatives,and nanocomposites,are described.Gas pressure,light wavelengths,relative humidity,cooling,and other environmental factors are important for improved postharvest storage.These elicitors maintain redox status by inhibiting the generation of reactive oxygen species(ROS)or their lysis.Growth regulators,including abscisic acid,auxin,brassinosteroids,jasmonic acid,and salicylic acid,are important for the regulation of ripening.Mechanical injuries,ionic imbalances,temperature variations,and tissue dehydration can occur irrespective of ripening cate-gories.The use of synthetic physiochemically active compounds is discussed in terms of physiological,metabolic,cellular,and molecular functions.Ethylene-induced autocatalytic processes,antioxidant cascades,epigenetic regulation,and homeodomain gene expression are discussed.Sugar–acid metabolism,dissolution of the cell wall,and direct or indirect production of secondary metabolites related to postharvest storage are mentioned regarding chilling storage.Elicitors and agrochemicals that trigger plant defense to increase secondary metabolite production are discussed for reducing fruit senescence during postharvest storage.

First molecular identification of Leishmania species in a new endemic area of cutaneous leishmaniasis in Lorestan,Iran

Objective:To identify Leishman{u using PCR.Methods:This studs was conducted from April2009 to March 2011 in order to identify Leishmania species in a new endemic area of CL in Lorestan.Iran.Samples were taken from 62 patients that referred to the health centers in different cities of Lorestan province,the presence of Leishmcania was confirmed using direct smear and then grown in NNN media and mass cultured in RPM!1640 medium supplemented with 10%heat-inactivated fetal bovine serum.DNA was extracted from cultured promastigotes and used in P15-PCR.Results:45(72.6%)samples out of 62 showed a hand in the range of 485 hp and 17(27.4%)with a hand in the range of 626 hp which were similar to standard strains of Leichmania tropica(L.tropical and Leishnrania major(L.major),respectively.50(65.80%)of samples were collected from people with no history of travel in at least a year prior to the onset which shows that indigenous source of infection.Conclusions:Since the vector and reservoir of the two species are different.so precise and extensive control and prevention methods should be designed and earned out.

Hepatitis B virus genotypes in precision medicine of hepatitis Brelated hepatocellular carcinoma:Where we are now

Hepatitis B virus(HBV)infection is a major player in chronic hepatitis B that may lead to the development of hepatocellular carcinoma(HCC).HBV genetics are diverse where it is classified into at least 9 genotypes(A to I)and 1 putative genotype(J),each with specific geographical distribution and possible different clinical outcomes in the patient.This diversity may be associated with the precision medicine for HBV-related HCC and the success of therapeutical approaches against HCC,related to different pathogenicity of the virus and host response.This Editorial discusses recent updates on whether the classification of HBV genetic diversity is still valid in terms of viral oncogenicity to the HCC and its precision medicine,in addition to the recent advances in cellular and molecular biology technologies.

Exploring the impact of hepatitis B immunoglobulin and antiviral interventions to reduce vertical transmission of hepatitis B virus

Hepatitis B virus(HBV)infection is a major public health burden.In HBV endemic regions,high prevalence is also correlated with the infections acquired in infancy through perinatal transmission or early childhood exposure to HBV,the socalled mother-to-child transmission(MTCT).Children who are infected with HBV at a young age are at higher risk of developing chronic HBV infection than those infected as adults,which may lead to worse clinical outcome.To reduce the incidence of HBV MTCT,several interventions for the infants or the mothers,or both,are already carried out.This review explores the newest information and approaches available in literature regarding HBV MTCT prevalence and its challenges,especially in high HBV endemic countries.This covers HBV screening in pregnant women,prenatal intervention,infant immunoprophylaxis,and postvaccination serological testing for children.

Effects of Imazethapyr-Based Herbicide Formulation in the Zebrafish (Danio rerio) Hepatocyte Cell Line (ZF-L): Cytotoxicity and Oxidative Stress

Seizures of agrochemical formulations have increased in Brazil and Rio Grande do Sul is among the Brazilian states with the highest number of seizures of these products obtained illicitly. The use of illicit formulations can cause significant harm to agricultural production, the environment, and non-target species. This study evaluated the cytotoxicity and oxidative stress of a seized formulation containing the herbicide imazethapyr (IMZT). Characterization of the herbicide included gas chromatography-mass spectrometry (GC-MS) and thermal analyses (thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC)). Hemolytic and cytotoxicity assays in ZF-L hepatic cells showed IC50 values of 12.75 µg/mL, 3.01 µg/mL, 2.67 µg/mL, and 1.61 µg/mL for erythrocytes, [3(4,5-dimethyl)-2 bromide-5 diphenyl tetrazolium] (MTT), neutral red (NR), and lactate dehydrogenase (LDH) assays, respectively. The median IC50 of 2.84 µg/mL was used in oxidative stress assays, revealing increased reactive oxygen species (ROS) production, reduced total sulfhydryl content, and decreased superoxide dismutase (SOD) and catalase (CAT) activity. This study is the first to report in vitro oxidative stress induced by IMZT in the ZF-L cell line, emphasizing the importance of in vitro assays for assessing the toxic effects of seized agrochemicals on human health and the environment.

Biofilm formation in clinical isolates of nosocomial Acinetobacter baumannii and its relationship with multidrug resistance

Objective: To check biofilm formation by Acinetobacter baumannii(A. baumannii)clinical isolates and show their susceptibility to different antibiotics and investigate a possible link between establishment of biofilm and multidrug resistance.Methods: This study was performed on clinical samples collected from patients with nosocomial infections in three hospitals of Tehran. Samples were initially screened by culture and biochemical tests for the presence of different species of Acinetobacter. Identifications were further confirmed by PCR assays. Their susceptibilities to 11 antibiotics of different classes were determined by disc diffusion method according to Clinical and Laboratory Standards Institute guidelines. The ability to produce biofilm was investigated using methods: culture on Congo red agar, microtiter plate, and test tube method.Results: From the overall clinical samples, 156 specimens were confirmed to contain A. baumannii. The bacteria were highly resistant to most antibiotics except polymyxin B.Of these isolates, 10.26% were able to produce biofilms as shown on Congo red agar.However, the percentage of bacteria with positive biofilm in test tube, standard microtiter plate, and modified microtiter plate assays were 48.72%, 66.66%, and 73.72%, respectively. At least 92% of the biofilm forming isolates were multidrug resistant.Conclusions: Since most of the multidrug resistant strains produce biofilm, it seems necessary to provide continuous monitoring and determination of antibiotic susceptibility of clinical A. baumannii. This would help to select the most appropriate antibiotic for treatment.

Detection of k-ras gene point mutation in fine needle aspiration and pancreatic juice by sequence special primer method and its clinical significance

INTRODUCTIONThe point mutation rate of k-ras gene at codon 12 inpancreatic adenocarcinoma is reported to be as highas 90%,and with no mutations in normalpancreas tissues or other pancreatic disorders.Wehave detected the presence of k-ras gene

Syzygium cumini(L.)Skeels:A review of its phytochemical constituents and traditional uses

Syzygium cumini(S.cumini)(L.) Skeels(jambolan) is one of the widely used medicinal plants in the treatment of various diseases in particular diabetes.The present review has been primed to describe the existing data on the information on botany,phytochemical constituents,traditional uses and pharmacological actions of 5.cumini(L.) Skeels(jambolan).Electronic database search was conducted with the search terms of Eugenia jambolana,S.cumini,jambolan,common plum and java plum.The plant has been viewed as an antidiabetic plant since it became commercially available several decades ago.During last four decades,numerous folk medicine and scientific reports on the antidiabetic effects of this plant have been cited in the literature.The plant is rich in compounds containing anthocyanins,glucoside,ellagic acid,isoquercetin,kaemferol and myrecetin.The seeds are claimed to contain alkaloid,jambosine,and glycoside jambolin or antimellin,which halts the diastalic conversion of starch into sugar.The vast number of literatures found in the database revealed that the extracts of different parts of jambolan showed significant pharmacological actions.We suggest that there is a need for further investigation to isolate active principles which confer the pharmacological action.Hence identification of such active compounds is useful for producing safer drugs in the treatment of various ailments including diabetes.

A duplex PCR for the rapid and simultaneous detection of Brucella spp.in human blood samples

Objective:To design a duplex PCR for rapid and simultaneous detection of Brucella species, in human blood samples.Methods:Fifty-two peripheral bloods samples were collected from suspicious patients with brucellosis.Following DNA extraction,PCR assay were performed, using three primers that could simultaneously identify and differentiate three major species of pathogenic Brucella in humans and animals.Results:Of the 52 peripheral bloods samples tested, 25 sample(48%) showed positive reactions in PCR.Twelve samples were positive for Brucella abortus(B.abortus)(23%,13 for Brucella melUensis(B.melUensis)(25%) and 0 for Brucella ovis (6.ovis)(Ow.Conclusions:This work de=monstrates dial in case where specific primers were utilized,duplex PCR has proved to be a simple,fast,and relatively inexpensive method for simultaneous detection of important species of Brucella in clinical samples.

Antibiotic resistance and carriage class 1 and 2 integrons in clinical isolates of Acinetobacter baumannii from Tehran,Iran

Objective:To investigate antibiotic resistance and carriage class 1 and 2 integrons in clinical isolates of Acinetobacter baumannii(A.baumannii)from Tehran,Iran.Methods:Antimicrobial susceptibility testing was performed according to the Clinical and Laboratory Standards Institute.The presence of integrons was investigated by PCR using specific primers.Results:Among isolated A.baumannii strains,82%were multidrug resistant,27 samples(54%)were resistant to three or more than three antibiotics and 16 samples(32%)showed resistance to two antibiotics.Integrons were detected from 44 of 50 isolates(88%),with classes 1 and 2 being observed in 42%(21/50)and 82%(41/50)of isolates,respectively.Integron-positive A.baumannii isolates showed higher antibiotic resistance than integron-negative isolates and all showed a multidrug-resistant phenotype.Conclusions:Our findings show that classes 1 and 2 integrons,and especially classes 2 integrons are widely disseminated among A.baumannii strains isolated from Tehran and these structures are playing a major role in the acquisition of multidrug resistance in these strains.So monitoring of drug resistance with investigating carriage class 1 and 2 integrons is very important to plan specific infection control measures due to multidrug resistance A.baumannii in Iran hospitals.

Simultaneous detection and differentiates of Brucella abortus and Brucella melitensis by combinatorial PCR

Objective:To evaluate simultaneous detection and differentiates of Brucella abortus(B.abortus) and Brucella melitensis(B.melitensis) through the combinatorial POR method.Methods:This study was designed using three primers that could simultaneously identify and differentiate two major species of pathogenic Brucella in humans and animals.Identification and differentiation of each species using the size of the PCR product were determined.To determine the specifieity of the method,bacteria close to the genus Brucella were used.Finally,to confirm PCR products.In addition to the products sequence,RFLP was performed on PCR products using restriction enzymes.Results:The method of optimized combinatorial PCR in this study could simultaneously detect and differentiate B.abortus and B.melitensis with high specificity and sensitivity in clinical samples.Differentiation of species is based on the resulting bands: therefore,the band 494 bp for B.abortus and 733 bp for B.melitensis were obtained.RFLP and sequencing results confirmed PCR results.Conclusions:The results of this study shows that without routine diagnostic methods such as culture and serology tests,using the molecular method of combinatorial PCR,important species of Brucella can be simultaneously identified and differentiated in clinical samples.

The aflatoxin B1 isolating potential of two lactic acid bacteria

Objective:To determine lactic acid bacteria's capability to enhance the process of binding and isolating aflatoxin B1 and to utilize such lactic acid bacteria as a food supplement or probiotic products for preventing absorption of aflatoxin Bl in human and animal bodies.Methods:In the present research,the bacteria were isolated from five different sources.For surveying the capability of the bacteria in isolating aflatoxin Bl,ELISA method was implemented,and for identifying the resultant strains through 16S rRNA sequencing method,universal primers were applied.Results:Among the strains which were isolated,two strains of Lactobacillus pentosus and Lactobacillus beveris exhibited the capability of absorbing and isolating aflatoxin Bl by respectively absorbing and discharging 17.4%and 34.7%of the aforementioned toxin existing in the experiment solution.Conclusions:Strains of Lactobacillus pentosus and Lactobacillus beveris were isolated from human feces and local milk samples,respectively.And both strains has the ability to isolate or bind with aflatoxin Bl.

Toll-like receptor 9 polymorphisms and Helicobacter pylori influence gene expression and risk of gastric carcinogenesis in the Brazilian population

BACKGROUND Toll-like receptors(TLRs)are the first line of host defense,and are involved in Helicobacter pylori(H.pylori)recognition and activation of both inflammatory and carcinogenic processes.The presence of single nucleotide polymorphisms(SNPs)in genes that activate the immune response may modulate the risk of precancerous lesions and gastric cancer(GC).Among them,Toll-like receptor 9(TLR9)polymorphisms have emerged with a risk factor of infectious diseases and cancer,however the studies are still inconclusive.AIM To evaluate whether TLR9 rs5743836 and rs187084 SNPs contribute to the risk of gastric carcinogenesis,and its influence on mRNA expression.METHODS A case-control study was conducted to evaluate two TLR9 SNPs(TLR9-1237 TCrs5743836 and TLR9-1486 CT-rs187084)in chronic gastritis(CG)and GC patients.A total of 609 DNA samples of peripheral blood[248 CG,161 GC,and 200 samples from healthy individuals(C)]were genotyped by polymerase chain reaction-restriction fragment length polymorphism.All samples were tested for the H.pylori infection using Hpx1 and Hpx2 primers.Quantitative polymerase chain reaction by TaqMan?assay was used to quantify TLR9 mRNA from fresh gastric tissues(48 GC,26 CG,and 14 C).RESULTS For TLR9-1237,the TC+CC or CC genotypes were associated with a higher risk of GC than C[recessive model odds ratio(OR)=5.01,95%confidence interval(CI):2.52-9.94,P<0.0001],and the CG(recessive model OR=4.63;95%CI:2.44-8.79,P<0.0001)groups.For TLR9-1486,an association between the CT+TT genotypes and increased risk of both GC(dominant model OR=2.72,95%CI:1.57-4.72,P<0.0001)and CG(dominant model OR=1.79,95%CI:1.15-2.79,P=0.0094)was observed when compared to the C group.Moreover,the presence of TLR9-1237 TC/CC+TLR9-1486 CC genotypes potentiate the risk for this neoplasm(OR=18.57;95%CI:5.06-68.15,P<0.0001).The TLR9 mRNA level was significantly higher in the GC group(RQ=9.24,P<0.0001)in relation to the CG group(RQ=1.55,P=0.0010)and normal mucosa(RQ=1.0).When the samples were grouped according to the polymorphic genotypes and the presence of H.pylori infection,an influence of TLR9-1237 TC+CC polymorphic genotypes(P=0.0083)and H.pylori infection(P<0.0001)was observed on the upregulation of mRNA expression.CONCLUSION Our findings show that TLR9 rs5743836 and rs187084 polymorphisms are associated with a higher risk of carcinogenesis gastric,and that TLR9 mRNA levels can be modulated by TLR9-1237 TC+CC variant genotypes and H.pylori infection.

Single nucleotide polymorphisms of OCTN1, OCTN2, and DLG5 genes in Greek patients with Crohn's disease

AIM: To validate novel single nucleotide polymorphisms (SNPs) in Greek patients with Crohn's disease (CD).METHODS: A total of 120 patients with CD, 85 patients with UC, and 100 unrelated healthy controls were genotyped. Genotyping was performed by allele-specific PCR or by PCR-RFLP analysis.RESULTS: Our results showed that the 1672T and -207C alleles were obviously over-represented in CD patients only (P＜0.01 and P＜0.05, respectively) compared to the control population. The G113A polymorphism was completely absent in our studied population. The odds ratio for the carriage of the TC haplotype was 2.21 for CD patients as compared with controls. Additionally, the frequency of the TC haplotype was increased in patients with ileocolitis or colitis, and was mainly associated with the fibrostenotic phenotype of the disease. Furthermore, when the TC haplotype was compared jointly with the carriage of at least one mutation of the NOD2/CARD15 gene, there was an increased risk for CD, but not for UC, compared to controls. Regarding the location of the disease, the concomitant presence of the TC haplotype and NOD2/CARD15 mutations was mainly associated with ileocolitis or ileitis. CONCLUSION: Collectively, our results suggest that the 1672T variant of the OCTN1 gene and the -207C variant of the OCTN2 gene represent risk factors for CD in the Greek population.

Self-Healing MXene-and Graphene-Based Composites:Properties and Applications

Today,self-healing graphene-and MXene-based composites have attracted researchers due to the increase in durability as well as the cost reduction in long-time applications.Different studies have focused on designing novel self-healing graphene-and MXenebased composites with enhanced sensitivity,stretchability,and flexibility as well as improved electrical conductivity,healing efficacy,mechanical properties,and energy conversion efficacy.These composites with self-healing properties can be employed in the field of wearable sensors,supercapacitors,anticorrosive coatings,electromagnetic interference shielding,electronic-skin,soft robotics,etc.However,it appears that more explorations are still needed to achieve composites with excellent arbitrary shape adaptability,suitable adhesiveness,ideal durability,high stretchability,immediate self-healing responsibility,and outstanding electromagnetic features.Besides,optimizing reaction/synthesis conditions and finding suitable strategies for functionalization/modification are crucial aspects that should be comprehensively investigated.MXenes and graphene exhibited superior electrochemical properties with abundant surface terminations and great surface area,which are important to evolve biomedical and sensing applications.However,flexibility and stretchability are important criteria that need to be improved for their future applications.Herein,the most recent advancements pertaining to the applications and properties of self-healing graphene-and MXene-based composites are deliberated,focusing on crucial challenges and future perspectives.

Evaluation of rabbit antibody response against 8 and 16 kDa recombinant subunits of antigen B from Echinococcus granulosus

Objective:To immunize rabbits with 12 and 16 kDa recombinant subunits of antigen B from Echinococcus granulosus(E.granulosus) and measuring polyclonal antibody and humoral immune response using ELISA and gel diffusion.Methods:Two mentioned antigens were cloned and expressed in expression vector and purified by affinity chromatography.Four young rabbits were selected and challenged intradermallt with yielded recombinant antigens.Rabbits’ sera were collected post infection and were tested using ELISA and gel diffusion for polyclonal antibody detection 10 days after last injection.Results:The specific antibody against the recombinant peptides was efficiently produced within 4 weeks post infection.Conclusions: Produced recombinants proteins could induce the immune response of the rabbits successfully. This process might improve the clarification of diagnosis and vaccination as regards hydatidosis.

Study of Helicobacter pylori genotype status in saliva,dental plaques,stool and gastric biopsy samples

AIM:To compare genotype of Helicobacter pylori(H.pylori) isolated from saliva,dental plaques,gastric biopsy,and stool of each patient in order to evaluate the mode of transmission of H.pylori infection.METHODS:This cross-sectional descriptive study was performed on 300 antral gastric biopsy,saliva,dental plaque and stool samples which were obtained from patients undergoing upper gastrointestinal tract endoscopy referred to endoscopy centre of Hajar hospital of Shahrekord,Iran from March 2010 to February 2011.Initially,H.pylori strains were identified by rapid urease test(RUT) and polymerase chain reaction(PCR) were applied to determine the presence of H.pylori(ureC) and for genotyping of voculating cytotoxin gene A(vacA) and cytotoxin associated gene A(cagA) genesin each specimen.Finally the data were analyzed by using statistical formulas such as Chi-square and Fisher's exact tests to find any significant relationship between these genes and patient's diseases.P < 0.05 was considered statistically significant,RESULTS:Of 300 gastric biopsy samples,77.66% were confirmed to be H.pylori positive by PCR assay while this bacterium were detected in 10.72% of saliva,71.67% of stool samples.We were not able to find it in dental plaque specimens.The prevalence of H.pylori was 90.47% among patients with peptic ulcer disease(PUD),80% among patients with gastric cancer,and 74.13% among patients with none ulcer dyspepsia(NUD) by PCR assay.The evaluation of vacA and cagA genes showed 6 differences between gastric biopsy and saliva specimens and 11 differences between gastric and stool specimens.94.42% of H.pylori positive specimens were cagA positive and all samples had amplified band both for vacA s and m regions.There was significant relationship between vacA s1a/m1a and PUD diseases(P = 0.04),s2/m2 genotype and NUD diseases(P = 0.05).No statically significant relationship was found between cagA status with clinical outcomes and vacA genotypes(P = 0.65).The evaluation of vacA and cagA genes showed 6 differences between gastric biopsy and saliva specimens and 11 differences between gastric and stool specimens,CONCLUSION:Regard to high similarity in genotype of H.pylori isolates from saliva,stomach and stool,this study support the idea which fecal-oral is the main route of H.pylori transmission and oral cavity may serve as a reservoir for H.pylori,however,remarkable genotype diversity among stomach,saliva and stool samples showed that more than one H.pylori genotype may exist in a same patient.

Midkine secretion protects Hep3B cells from cadmium induced cellular damage

AIM： To evaluate role Cadmium （Cd） exposure line Hep3B cells. of midkine secretion during in the human hepatocyte cell METHODS： Different dosages of Cd （0.5-1-5-10 μg/mL） were applied to Hep3B cells and their effects to apoptosis, lactate dehydrogenase （LDH） leakage and midkine secretion were evaluated as time dependent manner. Same experiments were repeated with exogenously applied midkine （250-5000 pg/mL） and/or 5 t^g/mL Cd. RESULTS： Cd exposure induced prominent apoptosis and LDH leakage beginning from lower dosages at the 48^th. Cd induced midkine secretion with higher dosages （P 〈 0.001）, （control, Cd 0.5-1-5-10μg/mL respectively： 1123 ± 73, 1157 ± 63, 1242 ± 90, 1886 ± 175, 1712 ± 166 pg/mL）. Exogenous 500-5000 pg/mL midkine application during 5 μg/mL Cd toxicity prevented caspase-3 activation （control, Cd toxicity, 250, 500, 1000, 2500, 5000 pg/mL midkine＋ Cd toxicity, respectively： 374 ± 64, 1786 ± 156, 1545 ± 179, 1203 ± 113, 974 ± 116, 646 ± 56, 556 ± 63 cfu） LDH leakage and cell death in Hep3B cells （P 〈 0.001）. CONCLUSION： Our results showed that midkine secretion from Hep3B cells during Cd exposure protects liver cells from Cd induced cellular damage. Midkine has anti-apoptotic and cytoprotective role during Cd toxicity. Further studies are needed to explain the mechanism of midkine secretion and cytoprotective role of midkine during Cd exposure. Midkine may be a promising theurapatic agent in different toxic hepatic diseases.