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The outbreak of 2019 Novel Coronavirus (2019-nCoV) has spread from Wuhan to the whole country. After the Spring Festival, workers will return to workplace and students will return to school. There is an increasing risk of 2019-nCoV cases being imported into provinces and cities. In order to promote the prevention and control of 2019-nCoV infection, reduce the risk of transmission in medical institutions, and ensure medical quality and medical safety, it is necessary to carry out the detection test of 2019-nCoV in biosafety class II laboratory. In order to achieve the goal of zero infection of the laboratory personnel, different preventive measures should be taken to assess the risk of the experimental activities.
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In order to promote the prevention and control of 2019-nCoV infection, reduce the risk of transmission in medical institutions, and ensure medical quality and medical safety, it is necessary to carry out the detection test of 2019-nCoV in biosafety class Ⅱ laboratory. In order to achieve the goal of zero infection of the laboratory personnel, different preventive measures should be taken to assess the risk of the experimental activities.
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Rapid diagnosis is important for the prevention and control of infectious disease.Point-of-care testing (POCT) technology is simple,mobile,rapid,sensitive and accurate.In recent years,POCT has been widely used in the detection of infectious agents.This article reviews the development of POCT in the diagnosis of infectious diseases,focusing on immuno-chromatographic technology,microfluidic chip technology and loop-mediated isothermal amplification (LAMP) technology.
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Objective@#To study the factors associated with efficacy of nucleos(t)ide analogues with sequential interferon in HBeAg negative chronic hepatitis B (CHB) patients.@*Methods@#HBeAg negative CHB patients with NA treatment received PEG-IFNα 2a 180 μg subcutaneously once weekly. NA was continually used with PEG-IFN 2a during the first 12 weeks. HBsAg level and HBV DNA load were observed in the sequential pre-treatment (baseline), 12th, 24th, 36th, 48th, 72nd and 96th weeks of sequential therapy in all patients.@*Results@#Of the 26 HBeAg negative CHB patients, 6 (23.1%) achieved HBsAg loss/seroconversion. The comparison between HBsAg loss/ seroconversion group and the group not achieved HBsAg loss/ seroconversion showed that the baseline HBsAg level in HBsAg loss/seroconversion group was 2.210 log10IU/ml, was lower than (t=-4.252, P=0.000) HBsAg-positive patients (3.385 log10IU/ml) in the groupp that not achieved HBsAg loss/seroconversion, the difference was statistically significant in HBsAg decreased at 72nd, 96th week in the course of sequential therapy (3.511 vs. 0.723log10IU/ml, 4.291 vs. 0.737 log10IU/ml) (t=6.712, P=0.000, t=13.319; P=0.000, 0.00); the difference was not statistically significant in age, gender and NA therapy time between the two groups; 12 (46.2%) of patients achieved sustained virologic response (SVR), the baseline of HBsAg level was 2.575 log10IU/ml, was less than (t=-4.319, P=0.000) the patients who did not achieve SVR (3.576 log10IU/ml), the difference was statistically significant in HBsAg decrease of each time in the course of sequential therapy between the two groups (0.612 vs. 0.088, 1.192 vs. 0.107, 1.566 vs. 0.167, 1.817 vs. 0.176, 2.424 vs. 0.193, 2.188 vs. 0.014, log10IU/ml) (t=3.109, 4.717, 4.500, 4.544, 5.560, 4.265, P=0.008, 0.010, 0.001, 0.001, 0.000, 0.003), the difference was not statistically significant in gender and NA therapy time, the difference was statistically significant in age (30.8 vs. 39.9 years) ( t=-2.219, P=0.038). Of 9 CHB patients whose baseline HBV DNA were positive, 5 patients (55.6%) had HBV DNA loss after sequential treatment, the patients whose HBV DNA loss with HBsAg decreased at 12th week in the course of sequential therapy, but there was no influence in the HBsAg decrease/loss after 12th week, the difference was not statistically significant in age, gender, NA therapy time, HBsAg and HBsAg decrease in the course of sequential therapy between HBV DNA-negative and HBV DNA-positive groups.@*Conclusions@#To HBeAg-negative CHB, patients with low baseline HBsAg (2 log10 IU/ml level) and significantly lower HBsAg decrease early were more likely to receive SVR. Among them, prolonged interferon therapy is easier to achieve HBsAg loss/seroconversion.
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Objective@#To investigate the differences in frequency and function of natural killer cells (NK) between chronic hepatitis B (CHB) and acute hepatitis B (AHB).@*Methods@#Patients with AHB and those with CHB in immune active (IA) phase were enrolled. The frequencies of NK, CD56dimNK, CD56brightNK and the expression of functional molecules IFNAR2 and NKp46 on the surface of NK cells were detected respectively among patients with CHB in IA phase, patients with AHB, and those recovered from AHB. At the same time, their correlations with ALT, HBV DNA and HBV markers were analyzed.@*Results@#Between IA and AHB, the frequencies of NK cells and NKp46dim NK cells in AHB cases were significantly lower than those in IA cases, but the frequency of NKp46high NK cells in AHB was higher than that in IA. For patients who recovered from AHB, the frequency of NK cells and NKp46dim NK cells increased; the varied ranges of frequencies of CD56dimNK, IFNAR2+ NK and NKp46+ NK cells were on the rise, while the frequency of NKp46high NK cells decreased after the recovery from AHB, and the varied ranges of CD56brightNK and IFNAR2MFI, NKp46MFI decreased. In AHB, HBVDNA loads were positively correlated with ALT levels. Before and after the recovery of AHB: ΔHBV DNA and ΔALT, Δ NK/LY (%) were positively correlated; ΔALT and ΔNKp46highNK/NK(%), ΔNKp46MFI, ΔIFNAR2MFI were positively correlated.@*Conclusions@#In CHB immune active phase, the activity of peripheral blood NK cells was too weak to remove the virus, but NK cells play an important role in eliminating the viruses and mediating liver tissue inflammation in AHB.
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Objective To compare the clinical feasibility of different methods for detecting Clostridium difficile (CD) .Methods The stool samples were collected from the patients with suspected antibiotic-related diarrhea during 2016 ,and the enzyme linked immunosorbent assay (ELISA) and identification medium culture method were used for detection .Then the sensitivity ,specificity and consistency were compared between the two methods .Results 29 cases of antigen-positive specimens were measured by ELISA ,including 25 cases of toxin-positive and 4 cases of toxin negative ;29 suspected strains were cultured by the selective identification medium ,which were identified by the mass spectrometry as 28 strains of Clostridium difficile and 1 strain of undetected species ;the sensitivity and specificity of ELISA method and identification culture medium method were 97% ,100% and 93% ,95% respectively ,both methods showed extremely good consistency (Kappa=0 .92) .Conclusion The ELISA method has the characteristics of fastness ,high efficiency ,time saving ,simple operation ,easy interpretation and so on ,and can rapidly and accurately screen CD related diarrhea diseases ;the medium culture method has a good specificity and can rapidly obtain the infectious strain ;their combined use can greatly increase the CD detection rate and has great help for late treatment .
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Objective@#To investigate the differences in function of plasmacytoid dendritic cells (pDC) and CD4+ T helper cells (CD4+ Th cells) between acute hepatitis B (AHB) and chronic hepatitis B (CHB).@*Methods@#In this study, patients with AHB and those with CHB in immune active (IA) phase were enrolled. The frequencies of pDC, CD86+ pDC, CD4+ T cells and their subsets, surface functional molecules were detected respectively among patients with chronic HBV infection in IA phase, patients with AHB, those recovered from AHB. Meanwhile, their correlations with ALT, HBV DNA and HBV markers were analyzed.@*Results@#The ALT level in AHB was significantly higher than that in IA, and inflammation was more obvious in AHB. Between IA and AHB, CD86+ pDC frequency and the mean fluorescence intensity of functional molecule CD86 (CD86MFI) were higher in IA than those in AHB, but the frequency of CD4+ T cells in AHB was higher than that in IA. For patients who got over AHB, the frequency of CD86+ pDC increased; Th1 were on the rise, while the frequencies of CD4+ T and Th2 decreased after the recovery of AHB, and Th2 / Th1 ratio decreased..In AHB, HBVDNA loads were positively correlated with ALT levels and Th2 frequencies.@*Conclusions@#In CHB immune active phase, CD86+ pDC with stimulating function played an important role, but the cellular immune response of CD4+ T cells decreased. In AHB inflammatory stage, CD4+ T cells played a strong cellular immune response, which result ed in viral clearance. Th2 cells regulation of CD4+ T cells played a dominant role, which was involved in the inflammatory response, and the cytotoxic role of Th1 cells during the recovery period was dominant, playing a strong cellular immune response, then the virus were completely eliminated.
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Objective@#To investigate the change of hepatitis B surface antibody (HBsAb) titer and its long-term protection and infection rates between 1 and 3-year-old children whose mothers were chronic hepatitis B pregnant woman with HBeAg positive and high viral load after successful blocking of mother-to-child transmission.@*Methods@#One-year-old children whose mothers were hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) positive, with HBV DNA≥106IU/ml were enrolled, then were followed up till 3 years old, and tested the five serological markers of hepatitis B and biochemical parameters at the age of one and three years respectively, and analyzed HBsAb titer, positive rate, negative rate and infection rate of 1 to 3-year-old children without enhanced vaccination; meanwhile, data of HBsAb titers at the age of 7 months were collected HBsAb titer, positive rate, and negative rate were analyzed.@*Results@#Totally 264 1-year-old children were enrolled into the study, including 178 children without enhanced vaccination between seven months and 1 year of age, and 114 children without enhanced vaccination between 1 year and 3 years of age. Our result showed that there were no infected children at the age between 1 and 3 years. HBsAb titer decreased from 7 months to 1 year old and dropped from 1 000 IU/L to 509.43 IU/L (P<0.05), and the antibody was still protective. From 1 year to 3 years old, HBsAb titer dropped from 466.72 IU/L to 67.3 IU/L (P< 0.05); at the age of 3 years, 60.52 % children were either weakly positive or negative, but still protective, but significantly less than those who had the reinforced vaccination. As a result , the children without the enhanced vaccination between 1 and 3 years of age were still at high risk.@*Conclusions@#If the antibody was protective at 7 months, children were not easily infected between 1 year and 3 years of age. At the age of 3, the antibody dropped to low or no responsive levels, and the children were still at high risk. It is necessary to take protective measures and supplement the vaccine.
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Objective@#To investigate the efficacy of 200IU hepatitis B immunoglobulin (HBIG) injection at 1 month after birth to interrupt the mother-to-children transmission (MTCT) of hepatitis B virus (HBV).@*Methods@#Infants born to mothers who were hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) positive, with HBV DNA load ≥1.0×106 IU/ml and who did not receive antiviral drug treatment during pregnancy, were randomly divided into 2 groups. Infants in the control group were treated with standard immunoprophylaxis: 200 IU HBIG and 10 μg recombinant hepatitis B vaccine injection within 2 h after birth and a vaccine booster at 1 and 6 months after birth. For infants in the HBIG group the standard immunoprophylaxis and an additional 200 IU HBIG were administered at 1 month. HBsAg, the antibody to HBsAg (anti-HBs), and HBV DNA load were measured at birth and after 7 months. later.Immunoprophylaxis failure was defined as the presence of HBV DNA and HBsAg positivity or the presence of HBV DNA and HBsAg negativity at 7 months.@*Results@#In this prospective cohort study, of the 280 infants enrolled, 14 infants (HBIG/control: 6/8) were lost to follow-up and 266 subjects (HBIG/control: 134/132) completed the 7-month study. The log10HBV DNA load of mothers in the HBIG group and control group were (7.31±0.66) log10IU/ml and (7.32±0.74) log10IU/ml, respectively (P=0.92). The MTCT rate of the two groups was similar (5.97% vs. 7.58%, P=0.63). At 7 months, the HBsAg positive rate and the level of anti-HBs in the two groups were 94.03%(126/134)vs. 91.67% (121/132) and 623.60±412.93 mIU/mL vs. 620.38±399.10 mIU/ml, respectively with no significant difference (P=0.48 and P=0.95, respectively). The log10 HBV DNA load of mothers in immunoprophylaxis failure group and success group was similar (P=0.09). The number of infants who were serum HBsAg positive and HBV DNA positive at birth in the immunoprophylaxis failure group were higher than those in the success group (100% and 100% vs. 35.89% and 31.85%, P<0.01, respectively). The serum HBsAg levels in infants at birth was the only independent relevant factor for HBV MTCT, with risk rates of 11.18 (95% Confidence interval (CI), 1.23-101.88), 352.00 (95%CI, 15.82-7833.20), and 968.00 (95%CI 81.35-11519.19) for HBsAg levels of 0.05-< 1, 1-< 10, and ≥ 10 IU/ml, respectively, compared to infants with HBsAg levels < 0.05 IU/ml.@*Conclusions@#Administering 200IU HBIG injection at 1 month did not reduce the risk of HBV MTCT.
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Objective To explore the related factors of hepatitis B virus(HBV) drug resistance and their clinical significance . Methods The retrospective analysis was performed on the dinical data in 153 cases of CHB in our hospital during 2011-2013 .The 8 viral drug resistance loci of 4 known nucleosides analogues lamivudine ,adefovir ester ,entecavir and telbivudine were performed the direct sequencing by using the double DNA end termination method(Sanger method);the related clinical factors of drug‐resist‐ant virus genotypes were statistically analyzed and the correlation between prognosis and drug resistance was analyzed .Results A‐mong 153 patients with CHB ,47 cases(29 .8% ) appeared virus genotype drug‐resistant ,the common sites were 204M‐I(18 cases) , 204M‐V(8 cases) ,followed by 181A‐T(10 cases) ,181A‐V(5 cases) ,then 180L‐M (14 cases) .The proportion of viral drug resist‐ance in the patients with family history ,HBeAg positive ,using the nucleotide analogues in the past and the baseline ALT≥5 times was significantly increased ,the incidence rates were 38 .8% ,34 .8% ,34 .6% and 50 .0% respectively .The multivariable COX re‐gression found that HBV‐DNA genetic drug resistance increased the risk of progression to cirrhosis of the liver ,the OR value was 4 .704(95% CI:1 .199 -18 .454) .Conclusion the Sanger method for direct sequencing is reliable and accurate method of HBV‐DNA genotype drug resistance .The proportion of viral drug resistance in the patients with a family history of hepatitis B ,HBeAg positive ,using nucleotide analogues and baseline ALT≥5 times is significantly increased;HBV‐DNA genotype drug resistance also increases the risk of progression to cirrhosis of the liver .
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Objective To establish and evaluate the review criteria of blood routine detection by the Sysmex XE‐5000 hematolo‐gy analyzer in our hospital for improving the effectiveness of the measurement results .Methods 606 blood samples were collected from the outpatients and inpatients during 2014 and performed the microscopic examination by 2 technologists‐in‐charge with the double‐blind method .The average value of these results was used as the judging criteria .According to the 41 items of rule formula‐ted by the international expert group and combining with the patients characteristics of infectious disease hospital and instrument features in this hospital ,20 items of review criteria for blood routine test of our hospital was formulated .Results With the micro‐scopic examination results as the criteria ,among 606 detected samples ,116 samples were in line with the rules and the review rate was 19 .14(116/606) ,in which the true positive rate was 6 .93% (42/606) ,the false positive rate was 12 .21% (74/606) ,the false negative rate was 2 .64% (16/606) ,the true negative rate was 78 .22% (474/606) ,the sensitivity was 72 .41% and the specificity was 86 .5% .The expected values of positive and negative results were 36 .21% and 96 .73% respectively .The total effective rate was 85 .15% .Conclusion The review criteria formulated by this laboratory are reasonable and effective ,which has an important significance in the clinical application for increasing the detection speed of blood routine ,improving the laboratory reports quality , without missed true blood patients and accurately providing the valuable information for clinicians .
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<p><b>OBJECTIVE</b>To investigate the correlation of CD(+)4 T lymphocyte count and prostaglandin E2 (PGE2) in gingival crevicular fluid (GCF) with periodontal status in HIV-positive patients with periodontitis.</p><p><b>METHODS</b>Twenty subjects were selected according to inclusion criteria. The plasmatic CD(+)4 T lymphocytes were counted. All the individuals were divided into three groups, group A (CD(+)4 T lymphocyte count < 200 cell/mm(3)), group B (200 cell/mm(3) ≤ CD(+)4 T lymphocyte count ≤ 500 cell/mm(3)) and group C (CD(+)4 T lymphocyte count > 500 cell/mm(3)). Periodontal indexes, including plaque index(PLI), bleeding index(BI), attachment level(AL) and probing depth(PD) were recorded.GCF samples were taken from 120 index teeth by means of sterile paper strips.GCF PGE2 levels were determined by radioimmunoassays. Mann-Whitney was used to compare the periodontal indexes and PGE2 levels among the three groups. Partial correlations and Spearman correlations were applied to analyze the correlation of CD(+)4 T lymphocytes count and PGE2 in gingival crevicular fluid with periodontal status.</p><p><b>RESULTS</b>BI value, PGE2 concentration and total PGE2 were 3.00(2.00), 90.75(30.60) µg/L, 447.58 (243.08) pg in group B, which were higher than those in group A[2.00(1.25), 79.75(30.50) µg/L and 339.52 (200.97) pg respectively] and group C[2.00(1.00), 73.38 (14.83) µg/L and 299.18 (108.33) pg respectively] (P < 0.0167). But the differences of PD and AL among the three groups were not significantly different(P > 0.0167). The correlations were observed between CD(+)4 T lymphocyte count and BI for the subpopulations with CD(+)4 T lymphocyte count <200 cells/mm(3) (r = 0.657, P < 0.05) and between 200-500 cells/mm(3) (r = -0.369, P < 0.05). PGE2 concentration was negatively correlated with BI, PD and AL (P < 0.05), and total PGE2 was positively correlated with PD and AL(P < 0.05).</p><p><b>CONCLUSIONS</b>There was an association between the periodontal status and CD(+)4 T lymphocyte count in HIV(+) patients.GCF PGE2 level was related to periodontal parameters including BI, PD and AL.</p>