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Influenza is an acute infectious disease of the respiratory tract and has posed a serious threat to human health. Vaccination is the most effective way to prevent influenza. The traditional inactivated or attenuated live vaccine based on chicken embryo has a long production cycle. Additionally, the surface antigens of vaccine strains and epidemic strains often do not match due to the continuous antigen drift or shift of influenza virus. As a result, the effectiveness of influenza vaccine is significantly reduced, so there is an urgent need to find a rapid and efficient influenza vaccine production strategy to overcome this limitation. In recent years, with the development of genetic engineering technology and various delivery systems, nucleic acid vaccine has been well developed, especially mRNA vaccine has attracted wide attention because of its simple production process and good safety. Therefore, mRNA-based vaccine immunization strategy provides a new direction for the prevention of influenza. This article reviews the molecular design, delivery system and research in influenza in the development of mRNA vaccine, in order to provide a reference basis for the research and development of influenza mRNA vaccine.
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Objective@#To develop a kind of quality control material which simulates clinical specimens for detecting plasma methylated Septin9 (mSEPT9) and investigate the performance for mSEPT9 detection in external quality assessment (EQA) of laboratories. @*Methods@#The cultured Hela and Jurkat cells, known to contain methylated and unmethylated Septin9 gene respectively, were cultured. The genomic DNA of the cells was collected and extracted, and detected by mSEPT9 kit. According to the Ct value, the genomic DNA was diluted into different concentrations of quality control materials with negative plasma. The homogeneity and stability of the quality control materials were evaluated. The panels consisted of 5 blindly coded samples were distributed to EQA participants and the results were summarized and evaluated. @*Results@#The purity and concentration of extracted genomic DNA met with the needs of use and could be used as quality control products for mSEPT9 detection. The homogeneity and stability met with the requirements of China National Accreditation Service for Conformity Assessment. Some of the participating laboratories occurred false positive results and false negative results, and a good linear correlation for the detected results (Ct values) was only observed in about 55.6% of the laboratories. Among the 9 participating laboratories, 7 laboratories (77.8%) performed well, 1 laboratory (11.1%) qualified, and 1 laboratory (11.1%) unqualified. @*Conclusion@#The quality control materials for mSEPT9 detection was successfully developed and the application in external quality assessment should be of great significance for evaluating and improving the detection ability of clinical laboratory.
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Objective@#To evaluate the external quality assessment (EQA) program for genotyping results of tacrolimus metabolism-related cytochrome P450 family 3 subfamily A member 5 ( CYP3A5 )using plasmid DNA constructed in vitro as quality control samples, discuss the problems in clinical laboratories enrolled in the program and improve the detection quality of CYP3A5 gene. @*Methods@#Recombinant plasmid carrying CYP3A5 *3 (rs776746) AA locus sequence was constructed as wild type sample and plasmid with CYP3A5 *3 GG mutation as mutant type sample. Heterozygous mutant samples were obtained by mixing the two plasmids with equal proportion. Recombinant plasmids DNA were used as the sample panel for EQA scheme. Participating laboratories were asked to test the samples using their routine methods and report the results before deadlines. The scores of each laboratory were calculated based on their results and the overall coincidence of different samples as well as the sensitivity and specificity of different methods. @*Results@#CYP3A5 *3 locus genotypes of the constructed plasmid were verified by Sanger sequencing. The results of 15 and 17 valid laboratories were received respectively in the two EQA programs. The total percentage of 93.33% (14/15) and 100% (17/17) of the laboratories submitted correct results for all the samples. The overall coincidence rates were 96% (72/75) and 100% (85/85) respectively. All the laboratories using digital FISH got full marks in two EQA schemes, while the coincidence rates were 90% (27/30) and 100% (40/40) for Sanger sequencing. @*Conclusion@#The recombinant plasmid DNA constructed in this study could effectively detect the performance of reagents with good clinical applicability. The results of EQA programs suggested that the overall accuracy rate of enrolled laboratories was high enough, while the performances in some laboratories still need to be improved. Quality controls in clinical laboratories were essential to assure the accuracy of results.
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Objective To evaluate the performance of MTHFR 677 genotyping external quality assessment ( EQA) program using plasmid DNA constructed in vitro as quality control samples and discuss the problems in clinical laboratories enrolled in the program .Methods Recombinant plasmid carrying MTHFR 677C locus sequence was constructed as wild type sample and plasmid with MTHFR 677T mutation was generated with site-directed mutagenesis as mutant type sample .Heterozygous mutant samples were obtained after equal proportion of the two plasmids .EQA scheme were held twice a year in 2016 and 2017, and sample panels contained 5 different samples using recombinant plasmid DNA containing all types of MTHFR 677 locus genotypes.Participating laboratories were asked to test samples using their routine methods and report the results before deadlines .26, 28, 52 and 56 effective reports were received respectively in the four EQA schemes .The scores of each lab were calculated based on their results and the overall compliance of different samples as well as the sensitivity and specificity of different methods were calculated using Microsoft Excel .Results MTHFR 677 locus genotypes of the constructed plasmid were verified by Sanger sequencing and there was no failure of sample detection in the four EQA schemes , which suggest that the plasmid has good clinical applicability .About 96.15％( 25/26 ) ,100％( 28/28 ) ,96.15％ (50/52)and 98.21％(55/56) of the laboratories submitted correct results for all samples in the four EQA schemes.The overall compliance rate were 99.23％ ( 129/130 ) , 100％( 140/140 ) ,96.92％ ( 252/260 ) and 98.93％(277/280) respectively.All laboratories using digital FISH and microarrays got full marks in four EQA schemes.The compliance rates for fluorescent PCR were 97.5％ ( 39/40 ) , 100％ ( 45/45 ) , 94.29％(66/70) and 100％ (95/95) respectively, while the rates were 100％ (20/20), 100％ (15/15), 90％(36/40) and 92.5％(37/40) for Sanger sequencing.Conclusions The recombinant plasmid DNA constructed in this study can effectively detect the performance of reagents with good clinical applicability.The results of EQA programs suggested that the overall accuracy rate of laboratories enrolled was high enough , while some laboratories′performance still needs to be improved .Quality controls in clinical laboratories were essential to assure the accuracy of results .
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Objective To investigate the relationship between anti-HCV antibody level and hepatitis C virus genotype in the patients.Methods Total of 603 anti-HCV positive serum samples were collected during 2013 to 2014 by retrospective research method.HCV RNA were detected in anti-HCV positive samples by repeat test and the genotype were detected in HCV RNA positive samples.The distribution of anti-HCV level in different hepatitis C genotype patients was analyzed and the body's response to viral antibodies and viral genotype correlation with anti-HCV concentration interquartile range was explored.Rates among genotype groups were compared using chi-square test.Results Totally 412 of 603 (68.33％) samples were anti-HCV positive by double reagent screening.174(42.3％) samples were detected as HCV RNA positive.The distributions of different anti-HCV level in different genotype patients were 1a(n =8) 1/8,1/8,4/8,2/8;1b(n =112)25.9％ (29/112),17.0％ (19/112),25.9％ (29/112),31.3％ (35/112);2a(n =14)3/14,4/14,5/14,2/14;3a(n =11)3/11,6/11,2/11,0/11;3b(n =16)4/16,11/16,1/16,0/16;6a(n =8)2/8,2/8,1/8,3/8 with anti-HCV concentration interquartile range respectively.The anti-HCV concentration distribution was different in patients with different HCV genotypes.The anti-HCV concentration distribution in patients of 1 b,2a and 6a genotypes were evently,while anti-HCV level was relatively high in 1a (13.65) and relatively low in 3b (8.77).There were differences in different genotypes of antibody concentrations (x2 =35.2,P ＜ 0.05).Conclusions There was correlation between anti-HCV level and HCV genotype.Because there were fewer cases in some genotypes,it was necessary to investigate more samples to corfirm the above results.
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Objective To investigate the prevalence of depression and its influencing factors in type 2 diabetes mellitus (T2DM)patients with long-term treatment of insulin pumps.Methods A total of 120 T2DM patients receiving long-term insulin pump treatment were enrolled,and 180 T2DM patients who received intensive insulin injection without pump in the same period were enrolled as controls.We assessed the depression in the two groups using Zung’s self rating scale.Results The depression scores of 32 patients with insulin pump were more than 0.50,and the depression prevalence of these patients was 26.67%,which was significantly lower than 48.89% in the control group (P < 0.05 ). The prevalence of moderate or severe depression was 3.33% and 1.67%, respectively,in the insulin pump group,which were considerably lower than the prevalence of 1 5.56% and 9.44%in the control group (P <0.05).Duration of DM and the number of complications might be independent risk factors of depression in DM patients with insulin pump.Conclusion Long-term use of insulin pump may effectively improve depressive state in T2DM patients through its optimal effect on maintaining blood glucose levels.
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BACKGROUND:Studies have reported that taurine has a certain therapeutic effect on the disease of various systems, such as nervous system, cardiovascular system, immune system and digestive system. The liver is the main place, also the important target organ, of taurine metabolism. Therefore, the relationship between taurine and hepatopathy has become a hot topic in recent years. OBJECTIVE:To investigate the influence of taurine on superoxide dismutase and malondialdehyde expression in the liver tissue of rat models of liver fibrosis induced by carbon tetrachloride. METHODS:Thirty male C57B/L rats of SPF grade were randomly and evenly divided into blank control, model and taurine groups. Rats in the blank control group were intraperitonealy injected with 100% peanut oil of 1 mL/kg, twice a week, in total 10 weeks. Rats in the model group were intraperitonealy injected with peanut oil of 1 mL/kg containing 20% carbon tetrachloride, twice a week, in total 10 weeks. Rats in the taurine group were intraperitonealy injected with peanut oil of 1mL/kg containing 20% carbon tetrachloride, twice a week, in total 10 weeks, and were intragastricaly administered taurine of 500 mg/kg per day starting from the 3rd week til the 10th week. RESULTS AND CONCLUSION:Compared with the blank control group, the serum levels of hyaluronic acid, laminin, typeⅢ procolagen, typeⅣ colagen, alanine aminotransferase, aspartate aminotransferase were significantly increased (P < 0.05), the level of superoxide dismutase in the liver tissue was lowered (P < 0.05), the level of malondialdehyde in liver tissue was significantly increased (P < 0.05), and liver index was increased (P < 0.05) in the model group. Pathological examination showed that there were necrosis of liver cels, fat vacuoles, fibrous tissue hyperplasia and inflammatory cel infiltration in the rats of the model group. Compared with the model group, the serum levels of hyaluronic acid, laminin, typeⅢ procolagen, typeⅣ colagen, alanine aminotransferase, aspartate aminotransferase were significantly lowered (P < 0.05), the level of superoxide dismutase in the liver tissue was significantly increased (P < 0.05), the level of malondialdehyde in the liver tissue was significantly lowered (P < 0.05), and liver index was significantly decreased (P < 0.05) in the taurine group. Pathological examination showed that there were no inflammatory cel infiltration, fat vacuoles, and fibrous tissue deposition in the liver tissue. The results indicate that taurine can decrease the contents of superoxide dismutase and malondialdehyde, and relieve the degree of liver fibrosis induced by carbon tetrachloridevia exerting its antioxidative effects.
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Decades researches of molecular oncology have found some moleculars which can promote tumor growth and help cancer surrive.This brings new hope for the treatment of tumors.Epidermal growth factor receptor (EGFR) is the first important molecular to inhibit tumor growth.Fifty percent triple negative breast cancer (TNBC) and inflammatory breast cancer (IBC) overexpress EGFR.EGFR and its downstream pathway can regulate epithelial-mesenchymal transition,promote tumor cell migration.EGFR therapy can improve the chemosensitivity of TNBC cells,apoptotic signal transduction occur.These studies suggest that the EGFR targeted therapy may have an important role in the treatment of TNBC and IBC.
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Benefit from the completion of human genome project and the rapid development of bioinformatics,the clinical molecular diagnostics continues to expand its scope of application which involves more widely use in infectious diseases,genetic diseases and tumors and other aspects.In the clinical molecular diagnostics,the application of international standard materials could effectively guarantee the quality of laboratory tests.This paper reviews the research progress in well-established international standard materials used for molecular diagnosis.
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A near infrared (NIR) method for the detection of illegal addition of sildenafil citrate in capsules of Chinese traditional medicine was constructed. The diffuse reflectance near infrared spectra were recorded using integrated sphere and fiber optics respectively. Using the spectra of the sildenafil citrate standard as Reference spectra, the threshold of the correlation coefficient was set up based on the minimum effective concentration of the sildenafil citrate. By comparing the threshold with the correlation coefficient between the tested spectra and the Reference spectra in the selected spectral range, the tested sample contains sildenafil citrate is determined. For the integrated sphere method, the spectral range of 6070-5800 cm~(-1) and 4170-4070 cm~(-1) were selected, and 70% was set as threshold. For the fiber method, 6070-5800 cm~(-1) and 65% were used. 79 and 247 batches of the Chinese traditional medicines contained sildenafil citrate collected from the Chinese market were used to validate the two methods separately. The accuracy of the integrated sphere method was 91.14%, and that of f iber method was 87.45%.
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Objective With the same times of injection to compare low-dose intradermal regimen with routine-dose intramuscular inoculation in revaccination of non-responders to hepatitis B vaccine. Methods 40 healthy non-responder children collected by screening were administrated a three-dose revaccination randomly by intramuscular or intradermal route (10 vs 2 g per dose), and regularly tested for serologic markers up to five years. By the end of follow-up, a booster dose (5 μg) was given to those who had lost anti-HBs of ≥10 mIU/mL (seroprotection) and anamnestic response was estimated thereafter. Results All 17 intramuscular and 22 of 23 intradermal children effected seroprotection after revaccination. Intradermal children lost seroprotection over time significantly rapider compared with intramuscular children (Log Rank test, P= 0.029). In year 5, 50% of intramuscular but only 18.2% of intradermal children still maintained seroprotection (P=0.075). 12-14 days after the booster dose, all the eight intramuscular children developed an anamnestic response with anti-HBs titer increasing greater, but two of the 18 intradermal children failed to mount seroprotective level. Conclusion Three-routine-dose intramuscular revaccination was significantly effective than low-dose intradermal one with the same times of injection, especially in long-term immunity. We recommend routine-dose intramuscular protocol in revaccination of non-responders.
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Objective In order to observe the efficacy of low-dose in the third injection of hepatitis B vaccine. Methods 126 children aged 5～9 years were enrolled in a double-blind, place-controlled and randomized field trial. They were randomly divided into 10μg and 20μg dose group, and were redivided into 2μg, 5μg, 10μg, 20μg or non-in jected subgroups when the third booster injections were given. Results During the 9 years follow-up, the differences of the anti-HBs levels(GMT) among the groups were not significant at every time (P ＞ 0. 05). The GMTs at the ninth year(T108) were 7. 0, 6. 4, 9.9, 6.1, 9.7, 5. 4 and 7.4, respectively (P＞ 0. 05). The HBV infection rates among the groups had no significant difference (P ＞ 0. 05). The protective rates in the groups were all higher than 75％ at T108. Conclusion According to the data, it can be concluded that the third injection with low-dose has no influence on the vaccine efficacy(either short-term or long-term efficacy).
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ObjectiveTo assess the efficacy and the immunological memory of plasma-derived hepatitis B vac- cine 11 years after the initial inoculation. Methods A randomized, double-blind and placebo-controlled trial design was used. Results The immunogenicity and protection rate of the vaccine were good 1 1 years after vaccination, how- ever, from 9 to 11 years after vaccination, the perscn year HBV infection rate showed no noticeable difference be- tween the vaccine group and placebo-controls. Furthermore, the immunological memory remained 11 years after in- oculation, but was significantly lower than that observed in the past 10 years. Conclusion Vaccine protection wanes over the years. More information is needed to define the appropriate time for vaccine booster doses.
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Objective To analyze the changes of insulin-like growth factor-1(IGF-1) serum level in diabetic lower extremity angiopathy,explore the role of IGF-1 in the pathogenesis of diabetic vascular complications,and provide basis for prevention and treatment of diabetic angiopathy.Methods We selected 74 type 2 diabetes mellitus(T2DM) patients as case group and 15 normal people as control group.All of the research subjects were measured in height,weight,blood pressure,blood glucose,blood lipid,fast insulin,and urinary albumin excretion rate(UAER).The serum IGF-1 level was measured with ELISA.Arteries of both lower extremities were measured with color Doppler flow image.Results ① The serum IGF-1 level was significantly higher in DM patients than in control group.② The serum IGF-1 level was significantly higher in diabetic patients with lower extremities arterial vasculopathy than diabetic patients without.The results showed a positive correlation between the serum IGF-1 level and severity of lower extremity arterial vasculopathy.③ There was a positive correlation between diabetic duration,UAER,total cholesterol(TC),lower density lipoprotein(LDL) in blood and lower extremity arterial vasculopathy in diabetic patients.Conclusion The serum IGF-1 level can reflect the severity of lower extremity arterial vasculopathy.The serum IGF-1 level may be a reference valuable in diagnosis of diabetic macroangiopathy and a new cure point for diabetic macroangiopathy.