Effects of potassium lactate on sensory attributes,bacterial community succession and biogenic amines formation in Rugao ham

To deepen the understanding in the effect of potassium lactate on the sensory quality and safety of Rugao ham,sensory attributes,physicochemical parameters,total volatile basic nitrogen(TVBN),microorganism community and biogenic amines of Rugao ham manufactured with different potassium lactate levels(0%,0.5%,1%,2%)were investigated;the relationship between microbial community and the formation of TVBN and biogenic amines was further evaluated.With the increase of potassium lactate from 0%to 2%,the increased sensory scores and the decreased total aerobic bacterial count and TVBN were observed;the abundance of Staphylococcus increased,while the content of Halomonas decreased.LDA effect size(LEf Se)and correlations analysis showed that Staphylococcus equorum and Lactobacillus fermentum could be the key species to improve sensory scores and decrease biogenic amines and TVBN.Metabolic pathway analysis further showed that amino acids metabolism and nitrogen metabolism were mainly involved in decreasing TVBN and biogenic amines in the treatment of 2%potassium lactate.

Cover Enhancement Method for Audio Steganography Based on Universal Adversarial Perturbations with Sample Diversification

Steganography techniques,such as audio steganography,have been widely used in covert communication.However,the deep neural network,especially the convolutional neural network(CNN),has greatly threatened the security of audio steganography.Besides,existing adversarial attacks-based countermeasures cannot provide general perturbation,and the trans-ferability against unknown steganography detection methods is weak.This paper proposes a cover enhancement method for audio steganography based on universal adversarial perturbations with sample diversification to address these issues.Universal adversarial perturbation is constructed by iteratively optimizing adversarial perturbation,which applies adversarial attack tech-niques,such as Deepfool.Moreover,the sample diversification strategy is designed to improve the transferability of adversarial perturbations in black-box attack scenarios,where two types of common audio-processing operations are considered,including noise addition and moving picture experts group audio layer III(MP3)compression.Furthermore,the perturbation ensemble method is applied to further improve the attacks’transferability by integrating perturbations of different detection networks with heterogeneous architec-tures.Consequently,the single universal adversarial perturbation can enhance different cover audios against a CNN-based detection network.Extensive experiments have been conducted,and the results demonstrate that the average missed-detection probabilities of the proposed method are higher than those of the state-of-the-art methods by 7.3%and 16.6%for known and unknown detection networks,respectively.It verifies the efficiency and transferability of the proposed methods for the cover enhancement of audio steganography.

基于VOSviewer软件的针灸治疗急性腰扭伤研究进展的可视化分析

目的探讨基于VOSviewer软件的针灸治疗急性腰扭伤研究进展的可视化分析。方法检索中国知网、万方、维普三大数据库,收集1990年1月至2021年12月有关针刺治疗急性腰扭伤的相关文献,通过VOSviewer软件,对发文量、发文期刊、主要研究者、主要穴位及施术方式等进行可视化分析。结果共纳入文献823篇,年度发文量在1995年达到51篇,之后总体呈缓慢下降趋势。发文量最多的期刊《针灸临床杂志》共发文65篇,核心作者吴耀持共发表7篇文章。主要穴位中“后溪穴”出现频数最高,为100次,施术方式频数最多的是“推拿”,为51次。结论运用新的干预方法刺激穴位或多学科联合进行急性腰扭伤的研究可能是未来的发展趋势。

Effect of transition on QOL among Chinese children who underwent liver transplantation more than 5 years earlier

Objective:This study was conducted to test the effect of transition on the quality of life(QOL)among Chinese children who underwent liver transplantation more than 5 years earlier.Methods:We delivered the pediatric quality of life inventory(Peds QL)3.0 transplant module before and after“transition”intervention.Results:In the first posttransition measurement(6 months after“transition”),the sections on“about my medicines I,”“my transplant and others,”“treatment anxiety,”“how do I look,”“communication,”and total score were significantly different from those of the pretransition stage(P<0.05).However,there was no significance in the QOL in the second posttransition measurement(1 year after“transition”;P>0.05).Conclusions:The short-term effect of transition was definite,while the long-term effect needs further evidence.

Liver transplantation for hepatocellular carcinoma beyond the Milan criteria: A review

Liver transplantation(LT) has been accepted as an effective therapy for hepatocellular carcinoma(HCC). The Milan criteria(MC) are widely used across the world to select LT candidates in HCC patients. However, the MC may be too strict because a substantial subset of patients who have HCC exceed the MC and who would benefit from LT may be unnecessarily excluded from the waiting list. In recent years, many extended criteria beyond the MC were raised, which were proved to be able to yield similar outcomes compared with those patients meeting the MC. Because the simple use of tumor size and number was insufficient to indicate HCC biological features and to predict the risk of tumor recurrence, some biological markers such as Alphafetoprotein, Des-Gamma-carboxy prothrombin and the neutrophil-to-lymphocyte ratio were useful in selecting LT candidates in HCC patients beyond the MC. For patients with advanced HCC, downstaging therapy is an effective way to reduce the tumor stage to fulfill the MC by using liver-directed therapy such as transarterial chemoembolization, radiofrequency ablation and percutaneous ethanol injection. This article reviews the recent advances in LT for HCC beyond the MC.

Risk factors for new onset diabetes mellitus after liver transplantation: A meta-analysis

AIM: To determine the risk factors for new-onset diabetes mellitus(NODM) after liver transplantation by conducting a systematic review and meta-analysis.METHODS: We electronically searched the databases of MEDLINE, EMBASE and the Cochrane Library from January 1980 to December 2013 to identify relevant studies reporting risk factors for NODM after liver transplantation. Two authors independently assessed the trials for inclusion and extracted the data. Discrepancies were resolved in consultation with a third reviewer. All statistical analyses were performed with the Rev Man5.0 software(The Cochrane Collaboration, Oxford, United Kingdom). Pooled odds ratios(OR) or weighted mean differences(WMD) with 95% confidence intervals(CIs) were calculated using either a fixed effects or a random effects model, based on the presence(I2 < 50%) or absence(I2 > 50%) of significant heterogeneity. RESULTS: Twenty studies with 4580 patients were included in the meta-analysis, all of which were retrospective. The meta-analysis identified the following significant risk factors: hepatitis C virus(HCV) infection(OR = 2.68; 95%CI: 1.92-3.72); a family history of diabetes(OR = 1.69, 95%CI: 1.09-2.63, P < 0.00001); male gender(OR = 1.53; 95%CI: 1.24-1.90; P < 0.0001); impaired fasting glucose(IFG; OR = 3.27; 95%CI: 1.84-5.81; P < 0.0001); a family history of diabetes(OR = 1.69; 95%CI: 1.09-2.63; P = 0.02); use of tacrolimus(OR = 1.34; 95%CI: 1.03-1.76; P = 0.03) and body mass index(BMI)(WMD = 1.19, 95%CI: 0.69-1.68, P < 0.00001). Other factors, such as hepatitis B virus infection and alcoholism, were not found to be associated with the incidence of NODM.CONCLUSION: The study showed that HCV infection, IFG, a family history of diabetes, male gender, tacrolimus and BMI are risk factors for NODM after liver transplantation.

Role of BMSCs in liver regeneration and metastasis after hepatectomy

Hepatocellular carcinoma(HCC),which develops from liver cirrhosis,is highly prevalent worldwide and is a malignancy that leads to liver failure and systemic metastasis.While surgery is the preferred treatment for HCC,intervention and liver transplantation are also treatment options for end-stage liver disease.However,the success of partial hepatectomy and intervention is hindered by the decompensation of liver function.Conversely,liver transplantation is difficult to carry out due to its high cost and the lack of donor organs.Fortunately,research into bone-marrow stromal cells(BMSCs)has opened a new door in this field.BMSCs are a type of stem cell with powerful proliferative and differential potential that represent an attractive tool for the establishment of successful stem cell-based therapy for liver diseases.A number of different stromal cells contribute to the therapeutic effects exerted by BMSCs because BMSCs can differentiate into functional hepatic cells and can produce a series of growth factors and cytokines capable of suppressing inflammatory responses,reducing hepatocyte apoptosis,reversing liver fibrosis and enhancing hepatocyte functionality.Additionally,it has been shown that BMSCs can increase the apoptosis rate of cancer cells and inhibit tumor metastasis in some microenvironments.This review focuses on BMSCs and their possible applications in liver regeneration and metastasis after hepatectomy.

Recent advances in prevention of hepatitis B recurrence after liver transplantation

Liver transplantation is the only effective treatment for hepatitis B virus(HBV)-related end-stage liver disease.However,without antiviral prophylaxis,the recurrence rate of hepatitis B is as high as 80%-100%,which leads to a 50% mortality rate in the first 2 years after liver transplantation.Combination therapy of hepatitis B immunoglobulin(HBIG) and lamivudine demonstrated a higher efficacy of prophylaxis and further reduced the rate of recurrence to < 10%.The strategy of HBIG combined with lamivudine has been the standard treatment in many centers.However,the high rate of lamivudine resistance and the many disadvantages of HBIG have compelled surgeons to reconsider the longterm efficacy of this strategy for the prevention of HBV reinfection.Recently,new nucleos(t)ide analogues,such as entecavir and tenofovir,have been approved as first-line monotherapies for the treatment of chronic hepatitis B infection.These antiviral medicines have replaced lamivudine as the first choice in the prevention of HBV recurrence after liver transplantation.Various therapies that are composed of entecavir,tenofovir,and lamivudine plus adefovir,with or without HBIG have been adopted in several liver transplant centers.This article reviews the recent advances in prophylaxis for the recurrence of hepatitis B after liver transplantation.

Liver transplantation for biliary atresia: A single-center study from China's Mainland

AIM: To summarize our single-center experience with liver transplantation(LT) for biliary atresia(BA).METHODS: From October 2006 to December 2012, 188 children with BA were analyzed retrospectively. The stage?Ⅰ?group(from October 2006 to December 2010) comprised the first 74 patients, and the stage Ⅱ group(from January 2011 to December 2012) comprised the remaining 114 patients. Finally, 123 liver transplants were performed in 122(64.9%) patients, whereas 66 patients did not undergo LT due to denial by their parents or lack of suitable liver grafts. The selection of graft types depended on the patients' clinical status and whether a suitable living donor was available. The characteristics of patients in stages?Ⅰ?and Ⅱ were described, and the surgical outcomes of LT recipients were compared between the two stages. The KaplanMeier method was used to estimate the cumulative patient and graft survival rates, and the equality of survival distributions was evaluated using the log-rank test.RESULTS: The 188 children consisted of 102 boysand 86 girls. Their ages ranged from 3 to 144 mo with a median of 8 mo. One hundred and fifteen(61.2%) patients were born in rural areas. Comparing stage?Ⅰ?and stage Ⅱ patients, the proportion of patients referred by pediatricians(43.2% vs 71.1%, respectively; P < 0.001) and the proportion of patients who previously received a Kasai procedure(KP)(32.4% vs 44.7%, respectively; P = 0.092) obviously increased, and significantly more parents were willing to treat their children with LT(73% vs 86%, respectively; P = 0.027). Grafts from living donors(102/122, 83.6%) were the most commonly used graft type. Surgical complications(16/25, 64.0%) were the main reason for posttransplant mortality. Among the living donor liver transplantation recipients(n = 102), the incidence of surgical complications was significantly reduced(34.1% vs 15.5%, respectively; P = 0.029) and survival rates of patients and grafts were greatly improved(81.8% vs 89.7%, respectively, at 1 year; 75.0% vs 87.8%, respectively, at 3 years; P = 0.107) from stage?Ⅰ?to stage Ⅱ.CONCLUSION: The status of surgical treatments for BA has been changing in China's Mainland. Favorable midterm outcomes after LT were achieved as centers gained greater technical experience.

Expression of IL-26 predicts prognosis of patients with hepatocellular carcinoma after surgical resection

Background: There is no data regarding prognostic impact of interleukin(IL)-26 on outcomes of patients with hepatocellular carcinoma(HCC). The present study aimed to evaluate the prognostic impact of IL-26 on HCC patients undergoing liver resection. Methods: From 2003 to 2008, 122 patients with HCC who received surgical curative resection were enrolled. Patients were stratified into IL-26-upper and-lower groups according to the median expression level from immunohistochemical staining of resected specimens. Prognostic impact of IL-26 was estimated using Kaplan–Meier curves. Univariate and multivariate analyses were performed to evaluate timedependent prognostic impact and independency of IL-26. Demographic and clinical factors that were associated with IL-26 were comprehensively identified. Results: Prognosis of the patients with high level of IL-26 revealed to be significantly unfavorable in both cumulative recurrence-free survival( P < 0.001) and overall survival( P = 0.002). Upper expression of IL-26(HR: 1.643;95% CI: 1.021 to 2.644;P = 0.041) and microvascular invasion(HR: 3.303;95% CI: 1.255 to 8.696;P = 0.016) were identified as significant independent prognostic factors for overall survival in the multivariable analysis. Conclusions: IL-26 is a novel prognostic factor for HCC after resection. Evaluation of IL-26 expression may be potentially valuable in clinical therapy when planning individualized follow-up schedule and evaluating candidates for prophylactic adjuvant treatment to prevent recurrence.

Impact of EBV infection and immune function assay for lymphoproliferative disorder in pediatric patients after liver transplantation: A single-center experience

Background:Post-transplant lymphoproliferative disorder(PTLD)is a lethal complication after pediatric liver transplantation,but information regarding risk factors for the development of PTLD remains unclear.This study was to identify characteristics and risk factors of PTLD.Methods:A total of 705 pediatric patients who underwent liver transplantation between January 2017 and October 2018 were studied.Impact of clinical characteristics and Epstein-Barr virus(EBV)infection on the development of PTLD was evaluated.In addition,ImmuKnow assay was adopted in partial patients to analyze the immune status.Results:Twenty-five(3.5%)patients suffered from PLTD with a median time of 6 months(3–14 months)after transplantation.Extremely high tacrolimus(TAC)level was found in 2 fatal cases at PTLD onset.EBV infection was found in 468(66.4%)patients.A higher peak EBV DNA loads(>9590 copies/mL)within 3 months was a significant indicator for the onset of PTLD.In addition,the ImmuKnow assay demonstrated that overall immune response was significantly lower in patients with EBV infection and PTLD(P<0.0001).The cumulative incidence of PTLD was also higher in patients with lower ATP value(≤187 ng/mL,P<0.05).Conclusions:A careful monitoring of EBV DNA loads and tacrolimus concentration might be supportive in prevention of PTLD in pediatric patients after liver transplantation.In addition,application of the ImmuKnow assay may provide guidance in reducing immunosuppressive agents in treatment of PTLD.

High mortality associated with gram-negative bacterial bloodstream infection in liver transplant recipients undergoing immunosuppression reduction

BACKGROUND Immunosuppression is an important factor in the incidence of infections in transplant recipient.Few studies are available on the management of immunosuppression(IS)treatment in the liver transplant(LT)recipients complicated with infection.The aim of this study is to describe our experience in the management of IS treatment during bacterial bloodstream infection(BSI)in LT recipients and assess the effect of temporary IS withdrawal on 30 d mortality of recipients presenting with severe infection.AIM To assess the effect of temporary IS withdrawal on 30 d mortality of LT recipients presenting with severe infection.METHODS A retrospective study was conducted with patients diagnosed with BSI after LT in the Department of Liver Surgery,Renji Hospital from January 1,2016 through December 31,2017.All recipients diagnosed with BSI after LT were included.Univariate and multivariate Cox regression analysis of risk factors for 30 d mortality was conducted in the LT recipients with Gram-negative bacterial(GNB)infection.RESULTS Seventy-four episodes of BSI were identified in 70 LT recipients,including 45 episodes of Gram-positive bacterial(GPB)infections in 42 patients and 29 episodes of GNB infections in 28 patients.Overall,IS reduction(at least 50%dose reduction or cessation of one or more immunosuppressive agent)was made in 28(41.2%)cases,specifically,in 5(11.9%)cases with GPB infections and 23(82.1%)cases with GNB infections.The 180 d all-cause mortality rate was 18.5%(13/70).The mortality rate in GNB group(39.3%,11/28)was significantly higher than that in GPB group(4.8%,2/42)(P=0.001).All the deaths in GNB group were attributed to worsening infection secondary to IS withdrawal,but the deaths in GPB group were all due to graft-versus-host disease.GNB group was associated with significantly higher incidence of intra-abdominal infection,IS reduction,and complete IS withdrawal than GPB group(P<0.05).Cox regression showed that rejection(adjusted hazard ratio 7.021,P=0.001)and complete IS withdrawal(adjusted hazard ratio 12.65,P=0.019)were independent risk factors for 30 d mortality in patients with GNB infections after LT.CONCLUSION IS reduction is more frequently associated with GNB infection than GPB infection in LT recipients.Complete IS withdrawal should be cautious due to increased risk of mortality in LT recipients complicated with BSI.

Hepatic artery reconstruction in pediatric liver transplantation:Experience from a single group

Background:The reconstruction of hepatic artery is a challenging part of the pediatric liver transplantation procedure.Hepatic artery thrombosis(HAT)and stenosis are complications which may result in ischemic biliary injury,causing early graft lost and even death.Methods:Two hundred and fifty-nine patients underwent liver transplantation in 2017 in a single liver transplantation group.Among them,225 patients were living donor liver transplantation(LDLT)and 34 deceased donor liver transplantation(DDLT).Results:In LDLT all reconstructions of hepatic artery were microsurgical,while in DDLT either microsurgical reconstruction or traditional continuous suture technique was done depending on different conditions.There were five(1.9%)HATs:four(4/34,11.8%)in DDLT(all whole liver grafts)and one(1/225,0.4%)in LDLT(P=0.001).Four HATs were managed conservatively using anticoagulation,and 1 accepted salvage surgery with re-anastomosis.Until now,3 HAT patients remain in good condition,whereas two developed biliary complications.One of them needed to be re-transplanted,and the other patient died due to biliary complications.Conclusions:Microsurgical technique significantly improves the reconstruction of hepatic artery in pediatric liver transplantation.The risk for arterial complications is higher in DDLT.Conservative therapy can achieve good outcome in selected HAT cases.

Effect of remote ischemic preconditioning among donors and recipients following pediatric liver transplantation:A randomized clinical trial

BACKGROUND Studies suggested that remote ischemic preconditioning(RIPC)may effectively lessen the harmful effects of ischemia reperfusion injury during organ transplantation surgery.AIM To investigate the protective effects of RIPC on living liver donors and recipients following pediatric liver transplantation.METHODS From January 2016 to January 2019 at Renji Hospital Affiliated with Shanghai Jiao Tong University School of Medicine,208 donors were recruited and randomly assigned to four groups:S-RIPC group(no intervention;n=55),D-RIPC group(donors received RIPC;n=51),R-RIPC group(recipients received RIPC,n=51)and DR-RIPC group(both donors and recipients received RIPC;n=51).We primarily evaluated postoperative liver function among donors and recipients and incidences of early allograft dysfunction,primary nonfunction and postoperative complications among recipients.RESULTS RIPC did not significantly improve alanine transaminase and aspartate aminotransferase levels among donors and recipients or decrease the incidences of early allograft dysfunction,primary nonfunction,and postoperative complications among recipients.Limited protective effects were observed,including a lower creatinine level in the D-RIPC group than in the S-RIPC group on postoperative day 0(P<0.05).However,no significant improvements were found in donors who received RIPC.Furthermore,RIPC had no effects on the overall survival of recipients.CONCLUSION The protective effects of RIPC were limited for recipients who received living liver transplantation,and no significant improvement of the prognosis was observed in recipients.

The utility of two-dimensional shear wave elastography and texture analysis for monitoring liver fibrosis in rat model

Background:Liver fibrosis is a common pathological change caused by a variety of etiologies.Early diagnosis and timely treatment can reverse or delay disease progression and improve the prognosis.This study aimed to assess the potential utility of two-dimensional shear wave elastography and texture analysis in dynamic monitoring of the progression of liver fibrosis in rat model.Methods:Twenty rats were divided into control group(n=4)and experimental groups(n=4 per group)with carbon tetrachloride administration for 2,3,4,and 6 weeks.The liver stiffness measurement was performed by two-dimensional shear wave elastography,while the optimal texture analysis subsets to distinguish fibrosis stage were generated by MaZda.The results of elastography and texture analysis were validated through comparing with histopathology.Results:Liver stiffness measurement was 6.09±0.31 kPa in the control group and 7.10±0.41 kPa,7.80±0.93 kPa,8.64±0.93 kPa,9.91±1.13 kPa in the carbon tetrachloride induced groups for 2,3,4,6 weeks,respectively(P<0.05).By texture analysis,histogram and co-occurrence matrix had the most frequency texture parameters in staging liver fibrosis.Receiver operating characteristic curve of liver elasticity showed that the sensitivity and specificity were 95.0%and 92.5%to discriminate liver fibrosis and non-fibrosis,respectively.In texture analysis,five optimal parameters were selected to classify liver fibrosis and non-fibrosis.Conclusions:Two-dimensional shear wave elastography showed potential applications for noninvasive monitoring of the progression of hepatic fibrosis,even in mild fibrosis.Texture analysis can further extract and quantify the texture features in ultrasonic image,which was a supplementary to further visual information and acquired high diagnostic accuracy for severe fibrosis.

Living-donor or deceased-donor liver transplantation for hepatic carcinoma:A case-matched comparison

AIM: To compare the surgical outcomes between living-donor and deceased-donor liver transplantation in patients with hepatic carcinoma.

Transarterial chemoembolization versus percutaneous microwave coagulation therapy for recurrent unresectable intrahepatic cholangiocarcinoma:Development of a prognostic nomogram

Background:Transarterial chemoembolization(TACE)and percutaneous microwave coagulation therapy(PMCT)are commonly used to treat intrahepatic recurrent liver cancers.However,there is no informa-tion regarding their effectiveness in patients with recurrent intrahepatic cholangiocarcinoma(ICC)after resection.Methods:A total of 275 patients with localized recurrent ICC who received either TACE(n=183)or PMCT(n=92)were studied.A propensity score matching analysis was performed to compare prognostic impact of TACE and PMCT.Prognostic factors for TACE and PMCT were identified respectively.Predictive nomograms for each TACE and PMCT were developed using the Cox independent prognostic factors and were validated in independent patient groups by receiver operating characteristic curves and area under curve values.Results:Both TACE and PMCT provided curativeness in partial patients(5-year overall survival:21.4%and 6.1%,respectively),but TACE provided better survival benefit in both overall patients(hazard ratio[HR]=0.71;95%confidence interval[CI]:0.50–0.97;P=0.034)and propensity score matching analysis(HR=0.69;95%CI:0.47–0.98;P=0.041).Independent prognostic factors for TACE were tumor size>5 cm,poor differentiation,and major resection,whereas poor differentiation,hepatitis B virus infection,cholelithiasis,and lymph node metastasis were identified for PMCT.Both predictive nomograms for TACE and PMCT were validated to be effective with area under curve values of 0.77 and 0.70,respectively.Conclusions:TACE provided better survival benefits compared to PMCT.However,there was a disparity in prognostic factors,suggesting evaluation of the two nomograms may be supportive in modality selection.Further prospective validation studies are required for the results to be applied in clinical medicine.

Diagnosis and outcomes of collateral arterial formation after irreversible early hepatic artery thrombosis in pediatric liver recipients

BACKGROUND： Early hepatic artery thrombosis （eHAT） has been recognized as an important cause of graft loss and mortality. However, the incidence, etiology and outcome are not dear, especially for children. The present study was to investigate the formation of collateral artery flow after irreversible eHAT and its impact on patient＇s prognosis. METHODS： We analyzed eHAT after liver transplantation in children from October 2006 to April 2015 in our center, illustrated the formation of collateral hepatic artery flow after irreversible eHAT and explored the diagnosis, complications, treatment and prognosis. The basic and follow-up ultrasonographic images were also compared. RESULTS： Of the 330 pediatric liver recipients, 22 （6.67%） de- veloped eHAT within 1 month. Revascularization attempts in- duding surgical thrombectomy, interventional radiology and conservational treatment （thrombolysis） were successful in 5 patients. Among the 17 patients who had irreversible eHAT, follow-up ultrasonography revealed that collateral artery flow was developed as early as 2 weeks after eHAT. Liver abscess and bile duct complication occurred secondary to eHAT in variable time. CONCLUSIONS： Collateral arterial formation is a compensatory adaptation to eHAT to supply blood to liver grafts. However, the severe bile duct damage secondary to eHAT is irreversible and retransplantation is unavoidable.

Quantitative Analysis of ATP Sulfurylase and Selenocysteine Methyltransferase Gene Expression in Different Organs of Tea Plant (<i>Camellia sinensis</i>)

Tea plant (Camellia sinensis) has unique biological features for the study of cellular and molecular mechanisms, an evergreen broad-leaved woody plant which can accumulate selenium in soil abundant of Selenium. Expression of the genes related to Selenium (Se) metabolism is an adaptation to the soil environment for a long period. The purpose of the present study was to explore if there exist differences of expression about these genes in tea plant between growing in Selenium-abundant and normal soil. A quantitative real-time reverse transcription polymerase chain reaction (Q-RT-PCR) assay was done for quantification of ATP sulfurylase (APS) and selenocysteine methyltransferase (SMT) mRNA normalized to Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene in tea plant. Young leaves, mature leaves and tender roots from tea plants growing in soil abundant of Selenium were respectively obtained from Shitai County, Anhui Province, and also the relevant materials of the selenium un-enriched tea plant planted at agricultural garden of Ahui Agriculture University were taken as control for real-time PCR analysis. The results showed that APS1, APS2 and SMT expression levels for either young or mature leaves in selenium-enriched tea plant were lower than that in ordinary (selenium un-enriched) tea plant. In contrast, the APS1, APS2 and SMT expression level of roots in selenium-enriched tea plant were all higher than that in ordinary tea plant. APS1 gene expression level of roots in selenium-enriched tea plant was about 1.6 times higher than that in the ordinary tea plant, APS2 gene expression level was about 4.8-fold higher than that in the ordinary tea plant, SMT gene expression level was about 3.3 times higher than that in the ordinary tea plant. Among various tissues of selenium-enriched tea plant, APS1 gene relative expression level of young leaves was similar to or slightly higher than mature leaves, and the one of roots was the lowest among them;APS2 gene relative expression level of young leaves was similar to or slightly higher than the roots, and the one of mature leaves was the lowest among them;SMT gene relative expression level of young leaves was similar to or slightly higher than mature leaves, and the one of roots was the highest among them. Our results suggest that there existed correlation between selenium and expression levels of these genes.

Inhibitory effects of antisense phosphorothioate oligodeoxynucleotides on pancreatic cancer cell Bxpc-3 telomerase activity and cell growth in vitro

AIM： To investigate the effect of telomerase hTERT gene antisense oligonucleotide （hTERT-ASO） on proliferation and telomerase activity of pancreatic cancer cell line Bxpc-3. METHODS： MTT assay was used to detect the effect of different doses of hTERT-ASO on proliferation of Bxpc-3 cell for different times. To study the anti-tumor activity, the cells were divided into there groups： Control group （pancreatic cancer cell Bxpc-3）; antisense oligonucleotide （hTERT-ASO） group; and nosense oligonucleotide group decorated with phosphorothioate. Telomerase activity was detected using TRAP-PCR-ELISA. Cell DNA distribution was examined using flow cytometry assay. Cell apoptosis was observed by transmission electron microscope in each group. RESULTS： After treatment with 6 mmollL hTERT- ASO, cell proliferation was inhibited in dose- and time- dependent manner. The telomerase activity decreased after treatment with hTERT-ASO for 72 h. Flow cytometry showed the cell number of G0/G1 phase increased from 2.7% to 14.7%, the cell number of S phase decreased from 72.7% to 51.0%, and a sub-G1 stage cell apoptosis peak appeared in front of G1 stage. CONCLUSION： Telomerase antisense oligodeoxy- nucleotide can inhibit the proliferation of pancreatic cancer cell line Bxpc-3 and decrease the telomerase activity and increase cell apoptosis rate in vitro.