Measurement agreement of 25-hydroxyvitamin D results derived from Roche immunoassay and liquid chromatography tandem mass spectrometry / 中华检验医学杂志

Objective:To evaluate the measurement agreement of Roche 25(OH)D immunoassay(evaluation method) with LC-MS/MS (reference method).Methods:A total of 909 residual serum samples from routine health check participants were collected from May to June in 2019. 25(OH)D concentrations were measured by evaluation method and LC-MS/MS, respectively. Passing-bablok regression, intraclass correlation coefficient (ICC), Bland Altman plots and Kappa test were used to analyze the consistency and bias on the results derived from the two measurement methods.Results:The 25(OH)D concentration derived from evaluation method was significantly different from those from LC-MS/MS method ( P<0.001). Slope of regression for evaluation method and LC-MS/MS was 0.962(95% CI 0.919-1.007), while intercept was -0.185 (95% CI -1.191-0.745). The ICC was 0.765 (95% CI 0.735-0.792). Altman plot showed that the average deviation between evaluation method and LC-MS/MS was -0.902 ng/ml (0.300%). The coincidence rate of evaluation method′s judgment of vitamin D sufficiency, insufficiency and deficiency with LC-MS/MS was 83.39%, and the weighted Kappa values was 0.790. Conclusion:Roche automatic 25(OH)D immunoassay shows acceptable correlation and agreement with LC-MS/MS, however, it is to note that the deviation between immunoassay and LC-MS/MS may lead to wrong judgment of vitamin D nutritional status. It is recommended that each laboratory should establish own corresponding reference values for 25(OH)D concentrations derived from these two methods.

The correlation between interleukin-28B gene polymorphism and its antiviral efficacy in patients with chronic hepatitis C infection / 中华检验医学杂志

Objective o evaluate the association between IL-28B ( rs12979860 ) polymorphism andantiviraltherapeutic effectbydetecting the genotype of interleukin-28B( IL-28 B) in patients with hepatitis C ( HCV ) .Methods Of total 1153 HCV patients, 303 diagnosed with CHC had been treated with pegylated interferon plus ribavirin for 24-48 weeks.IL-28B ( rs12979860 ) was genotyped by two-color fluorescent TaqMan assay.Results Among 1153 patients, CC, CT and TT genotype frequencies of IL-28B rs12979860 are 83.26%, 16.22%and 0.52%respectively.The results of HCV genotypingof 580 in 1153 cases, the frequencies of 1b, 2a and their non-1b/2a type are 63.45%, 35.00%and 1.55%respectively;In 303 CHC patients with clear medical history, the proportion of SVR was71.98% in patients with CC genotype and 16.90%in those with either the CT or TT genotypes.Logistic regression model was adopted to analyze the association of rs12979860 with SVR while adjusting for age, gender, viral load and HCV GT factors.Populations carrying combined genotype ( CT +TT) are making it harder to get SVR compared with those with CC genotype (OR, 95%CI:11.10,5.35-23.04;P<0.000 1).The percentages of SVR in HVC patients with 1b and 2a genotypeare 48.02% and 81.19% respectively.there is a statistically significant difference between these subgroups (χ2 =30.639,P<0.000 1).Conclusion IL-28B rs12979860 genotype is closely related to SVR in CHCpatients.Patients with CC genotype have a higher virus sustained response rate than those carrying CT or TT genotype.The SNP , rs12979860, might be applied as a predictor of clinical antiviral efficacy in the furture.

Better scientific administration on biosafety of a medical laboratory / 中华检验医学杂志

A lot of samples tested daily in clinical medical labs may contain one or more kinds of disease pathogeny which are potentially of biodamage toward surroundings and staff in the lab.Biosafety has since been an internationally important topic as public health control and the controllers' health under great negative impact.Following are some strategy which deserve more attention to strengthen our scientific management efficiency in biosafety of medical labs:(1)conduct bio-risk evaluation of the lab; (2) scientifically arrange layout and analysis proccsses in labs;(3 )providc with biosaf(e)ty instruments;(4)set up document series of biosafety; (5)strengthen safety protection on lab staffs;(6)handle samples and medical waste properly.

Requirement of diagnosis and tereatment for liver diseases and improvement of laboratory technology / 中华检验医学杂志

There are so many patients with different liver diseases in our country that the diagnosis and treatment of liver diseases directly relate to the people's healthy level.The requirement of diagnosis and treatment to clinical laboratories can be summarized as: ( 1 ) sensitivity,which means shortening window period;(2) specificity,which means various subtypes and variant can be detected;( 3 )speed,which means finding out the pathogen as soon as possible;(4)accuracy,which means providing physicians with accurate test reports.Detecting techniques,for instances,chemiluminescence,polymerase chain reaction and gene mutational site detecting method,were applied to the diagnosis and treatment of liver disease,thus improving the degree of precision,accuracy,sensitivity,specificity and detection speed and satisfying the requirement of diagnosis and treatment maximatily.

Isolation and identification of the viral pathogen in an incident with crowd fever of adults / 中华微生物学和免疫学杂志

Objective To isolate and identify the causative agent in an incident with crowd fever of adults in Henan province.Methods The cells was inoculated by the throat swabs of the patients and followed by neutralization assay and other molecular methods.Then indirect immunofluorescence assay was performed to detect the specific IgM/IgG antibodies against virus in the serum of the patients.Results We have isolated 2 strain adenoviruses from 10 swab samples,which were both identified as Ad11 by sequence analysis and neutralization test.6 of 10 samples were positive fer IgM specific for adenovirus and 3 positive for IgG.The remaining sample was negative for both.Conclusion The causative agent in this incident with crowd fever of adults was Ad11 in group B.

An outbreak of acute respiratory tract infection caused by Adenovirus in a military camp / 中华传染病杂志

Objective To describe the clinical characteristics and etiology diagnosis of an outbreak of acute respiratory tract infection in a military camp. Methods Two hundred and twenty-five cases were investigated using unified epidemiological questionnaires to describe the epidemiological characteristics. The etiological tests of 30 pharynx swab specimen were performed. The serum neutralized antibodies of 52 patients in acute and convalescence phases were detected by neutralization test. The patients and their close contacts were isolated. Air, stuffs and ground were all disinfected. Results Two hundred and twenty-five cases were distributed all the camp with obvious dormitory aggregation and the aggregation rate was 44. 9%. Among the 225 cases, all cases had fever and 161 (71.6%) had cough, 111 (49.3%) had pharyngalgia, 102 (45.3%) had headache, 31 (13.8%) had chest stuffy and 4(1. 8%) had dyspnea. Twenty (66. 7%) phargnxswab specimens turned to be adenorirus gene positive by polymerase chain reaction. Pharynx swab specimens were cultured in HepG2, Hela, RD and Vero cells, and 16 (53. 3%) presented with cytopathic changes. IgM antibody screening demonstrated that 24 cases were infected with Adenovirus, and neutralization test showed that antibodies increased ≥4 folds in paired sera from 28 cases. During 10 days after patient isolation and general disinfection, there was no new case of Adenovirus infection. Conclusions This outbreak of acute respiratory tract infection in a military camp was caused by Adenovirus, Timely andreasonable preventing measures can control the epidemic quickly.

Establishment of a PCR-product direct sequencing for the detection of HBV YMDD mutation / 中华检验医学杂志

Objective To develop an assay of PCR-produet direct sequencing to detect hepatitis B virus (HBV) YMDD mutation, and compare the results gained by the sequencing and traditional real-time fluorescent PCR assays. Methods Serum samples were collected from 103 patients with chronic hepatitis B. HBV DNA were extracted from sers. YMDD mutation was detected by a commercial real-time PCR assay. Meanwhile, HBV reverse transcriptase-encoding gene was amplified by a nested PCR assay. The PCR products were directly subjected to sequencing at two directions, and the sequencing results were analyzed by NTI program. Using Kappa test, comparison was made between the results of rtM204-site mutations obtained by the direct sequencing and YMDD mutations by the real-time fluorescent PCR. Results The direct sequencing assay proved to be highly effective with bread range of detection in viral load from 500 to 1010copies/ml. And it may simultaneously avoid inhibitory effect caused by high viral load. The coincidence rates between two assays were 100% for YIDD, 97. 1% for YVDD, 76. 2% for YIDD/YVDD coexistence (Kappa = 0. 853, P < 0. 01). Conclusions The direct sequencing assay for HBV drug-resistant mutation detection is highly sensitive with broad dynamic range. It has high coincidence rate with real-time fluorescent PCR assay with advantage of detecting YMDD, YIDD and YVDD mutations simultaneously.

The distribution and antimicrobial resistance tendency of pathogens associated with diarrhea in Beijing / 中华内科杂志

Objective To monitor the constituents and resistant tendency of bacterial pathogens isolated from diarrheal patients in our hospital form 1994 to 2005 to offer the basis for guiding epidemiologic study,vaccination research and clinical treatment. Methods Enteric pathogenic bacteria were cultured and identified to species,group and serotype with biochemical and serologic methods and the susceptibility of bacteria to antimicrobial agents were tested. Results Enteric pathogenic bacteria were isolated predominantly in male patients and mainly in children and youngsters. It reached a peak from July to September every year. Shigella spp.(75.11%) was the most frequendy isolated pathogens and followed by Vibrio spp.(12.7%),Salmonella spp.(6.28%),Aeromonas spp.(4.43%) and Escherichia coli(1.25%).During the period from 1994 to 2005,diarrheal pathogens had a trend of decrease especially Shigella spp.and Salmonella spp.. Of the 6329 isolates of Shigella spp., 75.62% was S. flexneri and S.soanei,S.dysenteriae and S. boydii constituted 23.98%,0.22% and 0.01% respectively.The sensitivity of different species,group or serotype to different antimicrobial agents was not the same.S.flexneri and Aeromonas spp. were highly resistant to most of antibiotics. However, S.sonnei and Vibrio spp.had good susceptibility to antibiotics tested except trimethoprim/sulfamethoxazole and ampicillin. Conclusion There are many species and serotypes of enteric pathogenic bacteria causing infective diarrhea and the distribution changes gradually in Beijing. The resistance rate of enteric pathogenic bacteria to antibiotics is not the same in different species and serotypes.so strict surveillance iS always needed.

The significant evaluation of eight biochemical items′ changing in liver disease / 中华检验医学杂志

Objective To discuss the diagnostic value of the examination of serum NAG, AFU, PAB, LAP, ASTm, GLDH,ADA and AFP in patients suffering from liver diseases.Methods Serum of 274 hepatitis cases and 30 healthy cases are examined with auto biochemical analyzer and analyzed statistically.Results The mean values of LAP, ASTm, GLDH, ADA and AFU in acute hepatitis patients are higher than health′s significantly, AUC of AFU,LAP and ASTm are 0.842,0.816 and 0.782 separately, positive rate of AFU,LAP and ASTm are 84.2%,95% and 80% separately; The mean values of ADA、AFU and NAG in liver cirrhosis patients are higher than health′s significantly while the mean value of PAB is lower significantly, AUC of ADA is 0.689, positive rate of ADA is 89.5%; The mean values of ADA and NAG in severe hepatitis patients are higher than health′s significantly while the mean values of PAB and AFU are lower significantly, AUC of PAB and AFU all is 0.861, positive rate of PAB and AFU is 100% and 52.1%; The mean values of LAP,AFP,NAG,ADA and AFU in liver cancer patients are higher than health′s significantly while the mean value of PAB is lower significantly, AUC of LAP and AFU is 0.697 and 0.653 separately, positive rate of LAP and AFU are 74% and 79.5% separately.Conclusions AFU、LAP and ASTm are valuable markers for diagnosing of acute hepatitis, ADA is a valuable marker for diagnosing of liver cirrhosis, PAB and AFU are valuable markers for diagnosing of severe hepatitis, LAP and AFP are valuable markers for diagnosing of liver cancer.

Clinical and pathological characteristics and pathogenesis of autoimmune hepatitis / 中华实验和临床病毒学杂志

<p><b>BACKGROUND</b>To explore the clinical and pathological characteristics and pathogenesis of autoimmunohepatitis (AIH).</p><p><b>METHODS</b>The serum and liver biopsy specimens and clinical data of 26 cases with patients with AIH were analyzed and scored according to the criteria of International autoimmune hepatitis (IAIHG, 1999). The changes of dendritic cells (DC) in the liver tissues were observed with a panel of DC markers (CD-80/B7-1, CD-86/B7-2, CD-1a and HLA-DR) and immunohistochemistry, and the activation of hepatic stellate cells (HSC) and the expression of TGF-alpha were also detected. Liver tissue specimens from 10 patients with chronic viral hepatitis B and C respectively and 5 normal liver specimens were chosen as controls.</p><p><b>RESULTS</b>Mean aggregate scores of 26 AIH cases, including 21 cases of type B (80.8%) and 5 cases of type C (19.2%), which were 18.6 +/- 1.4 and 19.1 +/- 2.1 respectively. There were significant differences between the type B and type C in the average age levels of serum ALT and AST, and alpha-Glo (P <0.001 or P< 0.01 or P <0.05). Histological features of all the AIH liver tissues showed the lesions of chronic active hepatitis such as interface hepatitis/piecemeal necrosis (100%), obvious lobular inflammation (type B 95.2%, type C 100%), bridging necrosis (57.1% type B, 80.0% type C, P<0.05), rosetting of liver cells (71.4% type B, 100% type C, P<0.01), central lobular confluent necrosis (33.3% type B, 80.0% type C, P<0.001), predominant plasmacytic infiltration (type B 95.2%, type C 20.0%, P<0.001). The rates of increased and concentrated DC in the portal and lobular areas, especially in the active lesions in type B and type C AIH were 85.7% (18/21) and 5/5 respectively. It was found that DC and lymphocytes surrounded the hepatocytes which partly expressed HLA-DR antigen, while there were no or a few HLA-DR positive hepatocytes in controls. Meanwhile, the number of alpha-SMA positive HSC and the expression of TGF- were obviously increased in AIH liver tissues.</p><p><b>CONCLUSIONS</b>Several clinical and pathological features of AIH were identified in this study. As an antigen-presenting cell, DC might play an important role in the pathogenesis of AIH. In China, sub-type B of AIH might be more frequent than sub-type C and there were differences in clinical aspects, serology and pathology between the two types.</p>

Treatment of severe viral hepatitis with artificial liver support system / 中华实验和临床病毒学杂志

<p><b>BACKGROUND</b>To investigate the method and therapeutic efficacy of artificial liver support system (ALSS) in treatment of severe viral hepatitis.</p><p><b>METHODS</b>A total of 83 patients including 66 with severe viral hepatitis were treated with ALSS using Baxter-550 artificial kidney and Biologic-DT system.</p><p><b>RESULTS</b>The levels of mean bilirubin, ALT, AST, BUN, Cr and endotoxin was significantly decreased after the treatment. Of the 66 patients?with severe viral hepatitis, 31(47.0%) had improvement in symptoms and 35 (53.0%) died or left hospital. In the control group,50(27.6%) out of the 181 had improvement in symptoms and 131(72.4%) died or left hospital.</p><p><b>CONCLUSIONS</b>ALSS could exert certain therapeutic effects on severe viral hepatitis.</p>

Dynamic studies on the serum cytokines level of different stage in the diagnosed SARS patients / 中华检验医学杂志

Objective To evaluate the clinical significance of the multiple cytokines in the serum of SARS patients and explore the rel ationship between the immune-reactivity and pathological damages. Methods 12 different serum cytokines have been detected in 4 groups ( inchoation, metaphase, convalescence of SARS patients and Healthy control) by using biochips technique(RANDOX) and to study the changes of each cytokine level in SARS patients. Results Compared with healthy group, IL-6 , IL-8 , IL-10 ,IFN? increased obviously and IL-1? ,IL-2,IL-4,VEGF,EGF,MCP-1, TNF?decreased obviously. Whereas IL-1?has no statistic changes among different stages of SARS. Conclusion There were obvious changes of multiple cytokines in different phase of SARS pathological process, especially in the early phase. It is further support the hypothesis that over-reaction of the immune system initiated the pathological injuries of the patients.

Dynamic analysis of anti-Cov antibody and SARS coronavirus in the plasma of 25 SARS patients / 解放军医学杂志

Objective To observe dynamic responsive regularity of specific IgM and IgG antibodies in SARS patients. Methods 145 specimens of plasma from 25 cases of clinically diagnosed SARS patients were examined in the study. ELISA was employed to detect IgM and IgG antibodies against SARS coronaviral antigens. Nested RT-PCR was used to qualitatively determine SARS coronavirus (SARS-CoV). Results 49.0% (71/145) and 54.5% (79/145) of the samples were positive for IgM and IgG antibodies, respectively. Both antibodies were found to be detected in 84.0% of the patients. The antibodies were found to be detectable from the 2nd to 4th week after the onset of disease in most patients, and there was a fendeney of sising in positive rate until the 5th week after the onset. Thereafter, the detectable rate of IgM antibody began to decline, while that of IgG antibody remained to rise. Positive rate for serum SARS-CoV was 15.8% (18/114) for all samples or 40.0% (10/25) of patients. Most virus-positive samples were those which were collected within 4 weeks after disease onset. Conclusions Anti-SARS-CoV IgM/IgG antibody and detection of virus in plasma could serve as practical diagnostic indicators for SARS. In most cases, when the serum was pasitive for antibody the serum virus positive rate would soon declined. However, concomitant existence of antibody and viral sequence in plasma was observed in a few patients.

Contribution of clinical laboratory in handling public health emergencies / 解放军医学杂志

Objective Accidental public health emergencies occurred frequently all over the world in the last decade. The problem of how to set up an effective system for prevention and control of infectious diseases has aroused the attention of all the nations worldwide. The present paper discussed the contributions of the clinical laboratory in hospital during public health emergencies by collecting samples, establishing and applying a rapid diagnosis platform for infectious pathogens, ensuring laboratory biosafety, and developing a strategy in case of emergency. These countermeasures may be helpful for the army to establish an effective system and mechanism in handling public health emergencies.

Application of PCR-RFLP technique on identification and genotyping of Brucella spp / 解放军医学杂志

Objective To amplify the 16S RNA fragments of 7 clinically isolated strains of Brucella spp. by PCR-RFLP technique, so as to provide experimental basis for the studies on diagnostics, genetics and epidemiology of Brucella spp. Methods According to the gene sequence of ATCC 25840 standard strain in GenBank, special primers for the 16S RNA conservative area in the Brucella spp. were designed. DNA extraction and PCR amplification of the 16S RNA fragments were performed with the 7 isolated strains. PCR products were then sequenced and RFLP analysis was conducted with appropriate restricted enzymes to study the homology and the mutation sites in those strains. Meanwhile, the clinical data of infected patients were retrospectively analyzed to evaluate the relationship between the clinical features and genotypes of Brucella infection. Results The amplified target fragments were about 1500bp in length and consistent with what was expected. The sequencing and homology analysis showed a 98.88% homology and 11 mutation sites among the 7 isolated strains. Four genotypes were identified by RFLP. Retrospective analysis of the clinical data indicated that no obvious relationship existed between the genotypes and the clinical features. Conclusions Amplifying 16S RNA fragments by PCR technique is a feasible method to make an early diagnosis of Brucella infection. The 7 clinically isolated strains are different in genotypes and 16S RNA fragment is a highly conservative fragment in bacterial genome with some mutations. The research provides evidence for the genetics and epidemiology of brucellosis.