Fast infectious diseases diagnostics based on microfuidic biochip system

Molecular diagnostics is one of the most important tools currently in use for clinical pathogen detection due to its high sensitivity,specificity,and low consume of sample and reagent is keyword to low cost molecular diagnostics.In this paper,a sensitive DNA isothermal amplifi-cation method for fast clinical infectious diseases diagnostics at aM concentrations of DNA was developed using a polycarbonate(PC)microfuidic chip.A portable confocal optical fuo-rescence detector was specifically developed for the microfuidic chip that was capable of highly sensitive real-time detection of amplified products for sequence-specific molecular identification near the optical diffraction limit with low background.The molecular diagnostics of Listeria monocytogenes with nucleic acid extracted from stool samples was performed at a minimum DNA template concentration of 3.65 aM,and a detection limit of less than five copies of genomic DNA.Contrast to the general polymerase chain reaction(PCR)at eppendorf(EP)tube,the detection time in our developed method was reduced from 1.5h to 45 min for multi-target parallel detection,the consume of sample and reagent was dropped from 25μL to 1.45μL.This novel microfuidic chip system and method can be used to develop a micro total analysis system as a clinically relevant pathogen molecular diagnostics method via the amplification of targets,with potential applications in biotechnology,medicine,and clinical molecular diagnostics.

Shear wave elastography may be sensitive and more precise than transient elastography in predicting significant fibrosis

BACKGROUND Noninvasive measurements including transient elastography(TE)and twodimensional shear wave elastography(SWE)have been used clinically instead of liver biopsy for regular assessment of liver fibrosis in chronic hepatitis B(CHB)patients.AIM To investigate the diagnostic efficiency of SWE compared to TE by assessing independent influencing factors and performance for diagnosing significant fibrosis based on our cohort of treatment-naive CHB patients.METHODS Fifty-four treatment-naive CHB patients who underwent liver biopsy to determine whether to initiate antiviral therapy were enrolled.SWE,TE,serum tests and liver biopsy were performed for all participants.The fibrosis-4 and aspartate aminotransferase to platelet ratio index scores were also calculated.Potential independent influencing factors on SWE and TE values were analyzed.Based on liver pathology results,the agreement and correlation were determined,and a comparison of the two methods was performed.RESULTS There were 27 cases(50%)of mild fibrosis(F0-F2)and 27(50%)cases of significant fibrosis(F3-F6);fibrosis was assessed with the Ishak scoring system.Multivariate linear regression analyses revealed that the fibrosis stage was the only factor that affected the SWE values(P<0.001),whereas the total bilirubin level(P=0.013)and fibrosis stage(P=0.037)were independent factors that affected TE values.Orthogonal partial least squares discriminant analysis showed that the number of independent factors(VIP>1)was higher for TE than SWE.Bland-Altman analysis showed satisfactory agreement between liver stiffness measurements(LSMs)of SWE and TE.Both SWE and TE could significantly discriminate significant fibrosis from mild fibrosis(P<0.001).SWE exhibited a higher correlation with LSMs of liver fibrosis than TE(r=0.65 and 0.50,P<0.001).The diagnostic performance of SWE was better than that of TE for significant fibrosis(F>2).The areas under the receiver operating characteristic curves of SWE and TE were 0.786 and 0.714,respectively.The optimal LSM cutoff values of SWE and TE were 9.05 kPa and 8.15 kPa,respectively.CONCLUSION Compared to the TE value,the SWE value was less affected by other factors.SWE may be more sensitive and precise than TE in predicting significant fibrosis(>F2)in CHB patients.

Synergistic interaction between thioredoxin inhibitor 1-methylpropyl 2-imidazolyl disul?de and sorafenib in liver cancer cells

Hepatocellular carcinoma(HCC)is a major health problem worldwide with high incidence and mortality rate[1,2].Proper treatments for HCC mainly include surgical resection,transcatheter arterial chemoembolization(TACE),radiofrequency ablation(RFA),and liver transplantation.However,many patients who are diagnosed with advanced HCC are not eligible for surgery,even the TACE and RFA does not necessarily have a good therapeutic effect.Until now,sorafenib is currently the first-line antitumor drug for the treatment of patients with advanced HCC.

Stereotactic body radiation therapy versus radiofrequency ablation in patients with small hepatocellular carcinoma:a systematic review and meta-analysis

Background:This study aimed to compare the clinical outcomes and toxicity between small hepatocellular carcinoma(HCC)patients treated with stereotactic body radiation therapy(SBRT)and those treated with radiofrequency ablation(RFA).Methods:We searched databases for relevant clinical studies.The primary outcomes of interest were overall survival(OS)at 1 and 2 years,freedom from local progression(FFLP)rate at 2 years,and complications.Results:Five cohorts from 5 retrospective studies and 4,814 patients with HCC were included.Pooled OS at 2 years was significantly lower for SBRT than for RFA[odds ratio(OR):0.63;95%confidence interval(CI):0.51-0.79;P<0.0001],but the pooled FFLP rate at 2 years was higher for SBRT than for RFA(OR:1.66;95%CI:1.05-2.61;P=0.03).In addition,there was no significant difference in the local and liver toxicities of the two treatments.The contradictory conclusion between the OS and FFLP outcome may be attributed to the difference in radiological dose and location,but there were no uniform criteria to illustrate the radiological dose and location in the included studies.Conclusions:SBRT had a higher local control ratio but poorer prognosis than RFA in patients with small HCC.The local toxicity was comparable in both treatments.Further trials should be designed with uniform standards for SBRT and RFA treatments.

Unsatisfying antiviral therapeutic effect in patients with mother-tochild transmissed chronic hepatitis B virus infection: a prospective multi-center clinical study

Background:Few data are available regarding the progression of liver disease and therapeutic efficacy in chronic hepatitis B(HBV)carriers infected by mother-to-child transmission(MTCT).This study aimed to investigate these two aspects by comparing the adult chronic HBV carriers in MTCT group with those in horizontal transmission group.Methods:The 683 adult chronic HBV patients qualified for liver biopsy including 191 with MTCT and 492 with horizontal transmission entered the multi-center prospective study from October 2013 to May 2016.Biopsy results from 217 patients at baseline and 78 weeks post antiviral therapy were collected.Results:Patients infected by MTCT were more likely to have e antigen positive(68.6%vs.58.2%,x^2=-2.491,P=0.012)than those with horizontal transmission.However,in patients with MTCT,levels of alkaline phosphatase(ALP)(P=0.031),Fibroscan(P=0.013),N-terminal propeptide of Type III procollagen(PIIINP)(P=0.014),and Laminin(LN)(P=0.006)were high,in contrast to the patients with horizontal transmission for whom the levels of albumin(ALB)(P=0.041),matrix metalloproteinase-3(MMP-3)(P=0.001)were high.The 47.2% of patients with MTCT and 36.8%of those with horizontal transmission had significant liver fibrosis(P=0.013).Following antiviral therapy for 78 weeks,21.2%and 38.0%patients with MTCT and horizontal transmission acquired hepatitis B e antigen(HBeAg)clearance,respectively(P=0.043),and the virological response rates were 54.7%and 74.1%in the MTCT and horizontal groups,respectively(P=0.005).MTCT was a risk factor for HBeAg clearance and virological response.Conclusion:Adult patients with MTCT were more prone to severe liver diseases,and the therapeutic efficacy was relatively poor,which underlined the importance of earlier,long-term treatment and interrupting perinatal transmission.

Gilbert syndrome combined with prolonged jaundice caused by contrast agent:Case report

This case highlights a patient with Gilbert syndrome who underwent endoscopic retrograde cholangiopancreatography(ERCP) with removal of bile duct stones, who then experienced an unexplained increase in bilirubin, with total bilirubin(TBIL) levels increasing from 159.5 μmol/L to 396.2 μmol/L and to a maximum of 502.8 μmol/L after 9 d. Following the decrease in the TBIL level, enhanced magnetic resonance cholangiopancreatography(MRCP) was performed to exclude any possible remaining choledocholithiasis. Nevertheless, the serum bilirubin level increased again, with TBIL levels rising from 455.7 μmol/L to 594.8 μmol/L and a maximum level of 660.3 μmol/L with no remaining bile duct stones. A liver biopsy showed severe bile duct cholestasis with no inflammation. Based on the exclusion of other potential causes of hyperbilirubinemia and the fact that both instances of increased bilirubin occurred after ERCP and MRCP, the contrast agents iopromide and gadoterate meglumine were suspected to be the causes of the hyperbilirubinemia. As of the writing of this report, the patient's bilirubin levels have spontaneously returned to baseline levels. In summary,ERCP and MRCP utilizing the contrast agents iopromide and gadoterate meglumine may possibly induce prolonged hyperbilirubinemia.

Serum HBV RNA as a predictor of virological response in treatmentnaive chronic HBeAg-positive HBV-infected patients with normal alanine aminotransferase

According to current guidelines,patients with chronic hepatitis B virus(HBV)infection whose HBVDNA is positive but whose alanine aminotransferase(ALT)levels are normal do not currently need antiviral treatment, However,studies have found that even chronic HBV infection'patients withnormal ALTmay,progress to cirrhosis or hepatocellular carcinoma(HCC).Furthermore,almost half of patients with normal ALT levels were found to have moderate to severe inflammation and/or significant fibrosis[2]Currently there were only a few studies on the treatment of chronic HBV patients with normal ALT levels,and the factors that affect their virological response(VR)are still unclear.An increasing number of studies have proven that serum HBV RNAcan be used as a non-invasive indicator to predict a VR,i3i In this study,weexplored the predictivevalue of HBV RNA in anticipating a VR in normal ALTchronic HBV patients.

Dysbiosis of Gut Microbiota Contributes to the Development of Diabetes Mellitus

Accumulating evidence has revealed that the composition of gut microbiota in patients with diabetes mellitus(DM)varies compared to those with healthy controls.The abnormal production and releases of metabolites derived from microbiota into the blood circulation contribute to the development of DM through the activation of multiple metabolic pathways,including trimethylamine Noxide,short-chain fatty acids,lipopolysaccharide,aromatic amino acids,and their related metabolites.Therefore,the modulation of gut microbiota through dietary intervention,probiotics,broad-spectrum antibiotics,and fecal microbiota transplantation could be a potential therapeutic approach for DM.This review mainly summarized the complicated interactions of gut microbiota through its metabolites with DM.

Significant Histologic Changes Are Not Rare in Treatment-naive Hepatitis B Patients with Normal Alanine Aminotransferase Level:A Meta-analysis

Background and Aims:Chronic hepatitis B is the main cause of liver cancer.However,the most neglected group has been treatment-naive chronic hepatitis B patients with normal alanine aminotransferase(ALT).People have tended to subjectively assume that the liver lesions of these patients are not serious and do not need antiviral treatment.However,the truth is not as optimistic as we thought.We aimed in this study to analyze the proportion of significant inflammation or fibrosis in aforementioned patients.Methods:Medline,Embase,and Cochrane Library were searched up to January 10th 2020,to identify studies of these patients with liver biopsy.The double arcsine method was used with a random-effect model to combine the proportion of significant inflammation or fibrosis.Potential heterogeneity was explored by subgroup analysis and meta-regression.Outcome of interests included the proportion of significant inflammation or fibrosis and cirrhosis.The secondary outcome was to find the risk factors of significant histological changes.Results:Nineteen eligible studies,with 2,771 participants,were included.The pooled proportion of significant inflammation or fibrosis was 35%[95%confidence interval(CI):27 to 43]and 30%(95%CI:25 to 36),respectively.The pooled proportion of cirrhosis was 3%[95%CI:1 to 5,(12 studies;1,755 participants)].In subgroup analysis,old age[vs.young(<40 years-old),44%vs.26%,p=0.012]was significantly associated with higher fibrosis stage as well as cirrhosis[vs.young(<40 years-old),4.8%vs.1.8%,p<0.001].Conclusions:About 1/3 of the treatment-naive chronic hepatitis B patients with normal ALT show significant histological changes,and some even have cirrhosis.

Hepatitis B core-related antigen reflects viral replication and protein production in chronic hepatitis B patients

Background:Hepatitis B core-related antigen(HBcrAg)is a promising disease-monitoring marker for chronic hepatitis B(CHB).We investigated correlations between HBcrAg with antiviral efficacy and virological and histological variables.Methods:One hundred and forty-five CHB patients from the mainland of China between August 2013 and September 2016 who underwent liver biopsy received entecavir therapy and had paired liver biopsy at 78 weeks.We analyzed correlations between HBcrAg and virological and histological variables in hepatitis B e antigen(HBeAg)-positive and HBeAg-negative patients.We also explored the predictors of HBeAg loss after 78 weeks of antiviral therapy.Pearson correlation analysis and logistic forward stepwise regression were the main statistic methods.Results:HBeAg-positive patients(n=93)had higher baseline HBcrAg(median 7.4 vs.5.3 log10 U/mL P<0.001)and greater HBcrAg declines(median 1.6 vs.0.9 log10 U/mL P=0.007)than HBeAg-negative patients after 78 weeks of therapy.At baseline,HBcrAg correlated with hepatitis B virus(HBV)DNA in both HBeAg-positive(r=0.641,P<0.001)and-negative patients(r=0.616,P<0.001),with hepatitis B surface antigen(HBsAg)in HBeAg-positive patients(r=0.495,P<0.001),but not with anti-hepatitis B virus core antibody(anti-HBc).Weak correlations existed between HBcrAg,histology activity index(HAI;r=0.232,P=0.025),and Ishak fibrosis score(r=-0.292,P=0.005)in HBeAg-positive patients.At 78 weeks,significant correlations existed only between HBcrAg and anti-HBc in HBeAg-positive(r=-0.263,P=0.014)and HBeAg-negative patients(r=-0.291,P=0.045).Decreased HBcrAg significantly correlated with reduced HBV DNA(r=0.366,P=0.001;r=0.626,P<0.001)and HBsAg(r=0.526,P=0.001;r=0.289,P=0.044)in HBeAg-positive and-negative patients,respectively,and with reduced HAI in HBeAg-positive patients(r=0.329,P=0.001).Patients with HBeAg loss(n=29)showed a larger reduction in HBcrAg than those without(median 2.3 vs.1.3 log10 U/mL,P=0.001).In multivariate analysis,decreased HBcrAg was an independent predictor of HBeAg loss(P=0.005).Conclusions:HBcrAg reflects viral replication and protein production.Decreased HBcrAg could predict HBeAg loss after antiviral therapy.Trial registration:Clinical Trials.gov:NCT01962155;https://www.clinicaltrials.gov/ct2/show/NCT01962155?term=NCT01962155&draw=2&rank=1.