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Objective:To establish a clinical laboratory diagnostic pathway for hepatitis C covering diagnosis, differential diagnosis, drug toxicity monitoring, and therapeutic and prognostic evaluation, and to explore a new teaching model for laboratory diagnostics based on the clinical laboratory diagnostic pathway.Methods:According to the clinical diagnosis and treatment guidelines for hepatitis C, laboratory testing strategies for different stages of diagnosis and treatment of the disease were formulated to establish a clinical laboratory diagnostic pathway for hepatitis C. The pathway was applied in the teaching for undergraduate medical students of the seven-year program of grade 2019 of The First Clinical College of Wuhan University, with those of grade 2018 as the control to receive traditional teaching. The teaching effect was compared through questionnaires and quizzes in class. The data were analyzed through the t test with the use of SPSS 19.0. Results:A clinical laboratory diagnostic pathway for hepatitis C recognized by clinicians was established, covering the entire process of clinical diagnosis, differential diagnosis, monitoring of drug side effects, and therapeutic and prognostic evaluation. The students of grade 2019 receiving the pathway-based teaching model had significant improvements in teaching quality evaluation indicators ( P<0.05), with the most marked improvement in "having mastered the key and difficult points of this lesson", with a score of (60.90±2.15) points for grade 2018 and (84.80±3.44) points for grade 2019. The total score for teaching evaluation was significantly higher in students of grade 2019 than in those of grade 2018 [(94.02±4.29) vs. (79.21±3.68)] points, P<0.05). Grade 2019 also had a significantly higher classroom quiz score than grade 2018 (94.60±5.63) vs. (78.10±4.92), P<0.01]. Conclusions:We established and applied a clinical laboratory diagnostic pathway of hepatitis C in the teaching model of laboratory diagnostics, which organically integrates laboratory diagnostics and clinical medicine, and significantly improves the quality of teaching.
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The gynecological tumours such as Breast cancer or female reproductive system tumors are a serious threat to female health. With the development of molecular diagnosis, the genomic changes of gynecological tumours have been revealed continuously, and the diagnosis and treatment modes of tumors have gradually changed. The detection of molecular targets which potentially participated in the transformation or progress of disease has become an important section of the management of female reproductive tumors, and accurate identification of molecular targets of tumors plays an important role in disease diagnosis, monitoring of metastasis, prediction of recurrence and treatment. This review briefly discusses the risk assessment, molecular typing, targeted therapy, toxic and side effects, and prognosis evaluation of breast and female reproductive system tumors by molecular diagnosis.
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BackgroundThe outbreak of COVID-2019 is becoming a global public health emergency. Although its basic clinical features have been reported, the dynamic characteristics of immune system in COVID-2019 patients, especially those critical patients with refractory hypoxemia, are not yet well understood. We aim to describe the dynamic characteristics of immune system in 3 critical patients with refractory hypoxemia, and discuss the relationship between hypoxemia severity and immune cell levels, and the changes of gut microbes of COVID-2019 patient. MethodsThis is a retrospective study from 3 patients with 2019-nCoV infection admitted to Renmin Hospital of Wuhan University, a COVID-2019 designated hospital in Wuhan, from January 31 to February 6, 2020. All patients were diagnosed and classified based on the Diagnosis and Treatment of New Coronavirus Pneumonia (6th edition) published by the National Health Commission of China4. We recorded the epidemiological history, demographic features, clinical characteristics, symptoms and signs, treatment and clinical outcome in detail. Blood samples were collected and we determined the expression levels of immune cells (CD3+ T cells, CD4+ T cells, CD8+ T cells, CD19+ B cells, and CD16+56+ NK cells) in different time points. Nanopore Targeted Sequencing was used to determine the alterations of gut microbiota homeostasis. ResultsApart from the clinical features described previously4, we found that four patients had decreased immune cells and refractory hypoxemia during the hospitalization, and the severity of hypoxemia was strongly correlated to the expression levels of immune cells. Additionally, we found that the proportion of probiotics was significantly reduced, such as Bifidobacterium, Lactobacillus, and Eubacterium, and the proportion of conditioned pathogenic bacteria was significantly increased, such as Corynebacterium of Actinobacteria and Ruthenibacterium of Firmicutes. Notably, all patients died. ConclusionsWe discussed the dynamic characteristics of host immune system and the imbalance of gut microbiota in 3 critical patients with COVID-2019. Hypoxemia severity was closely related with host immune cell levels, and the vicious circle between immune disorder and gut microbiota imbalance may be a high risk of fatal pneumonia. To the best of our knowledge, this is the first study which revealing that immunodepletion with refractory hypoxemia is a potential high risk subtype of COVID-2019 and the vicious circle between immune disorder and gut dysbiosis may be a high risk of fatal pneumonia.
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The ongoing novel coronavirus pneumonia COVID-19 outbreak in Wuhan, China, has engendered numerous cases of infection and death. COVID-19 diagnosis relies upon nucleic acid detection; however, current recommended methods exhibit high false-negative rates, low sensitivity, and cannot identify other respiratory virus infections, thereby resulting patient misdiagnosis and impeding epidemic containment. Combining the advantages of target amplification and long-read, real-time nanopore sequencing, we developed nanopore target sequencing (NTS) to detect SARS- CoV-2 and other respiratory viruses simultaneously within 6-10 h. Parallel testing with approved qPCR kits of SARS-CoV-2 and NTS using 61 nucleic acid samples from suspected COVID-19 cases confirmed that NTS identified more infected patients as positive, and could also monitor for mutated nucleic acid sequence or other respiratory virus infection in the test sample. NTS is thus suitable for contemporary COVID-19 diagnosis; moreover, this platform can be further extended for diagnosing other viruses or pathogens.
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Background2019-Novel coronavirus (2019-nCoV) outbreaks create challenges for hospital laboratories because thousands of samples must be evaluated each day. Sample types, interpretation methods, and corresponding laboratory standards must be established. The possibility of other infections should be assessed to provide a basis for clinical classification, isolation, and treatment. Accordingly, in the present study, we evaluated the testing methods for 2019-nCoV and co-infections. MethodsWe used a fluorescence-based quantitative PCR kit urgently distributed by the Chinese CDC to detect 8274 close contacts in the Wuhan region against two loci on the 2019-nCoV genome. We also analyzed 613 patients with fever who underwent multiple tests for 13 respiratory pathogens; 316 subjects were also tested for 2019-nCoV. FindingsAmong the 8274 subjects, 2745 (33.2%) had 2019-nCoV infection; 5277 (63.8%) subjects showed negative results in the 2019-nCoV nucleic acid test (non-2019-nCoV); and 252 cases (3.0%) because only one target was positive, the diagnosis was not definitive. Eleven patients who originally had only one positive target were re-examined a few days later; 9 patients (81.8%) were finally defined as 2019-nCoV-positive, and 2 (18.2%) were finally defined as negative. The positive rates of nCoV-NP and nCovORF1ab were 34.7% and 34.7%, respectively. nCoV-NP-positive only and nCovORF1ab-positive cases accounted for 1.5% and 1.5%, respectively. In the 316 patients with multiple respiratory pathogens, 104 were positive for 2019-nCov and 6/104 had co-infection with coronavirus (3/104), influenza A virus (2/104), rhinovirus (2/104), and influenza A H3N2 (1/104); the remaining 212 patients had influenza A virus (11/202), influenza A H3N2 (11/202), rhinovirus (10/202), respiratory syncytial virus (7/202), influenza B virus (6/202), metapneumovirus (4/202), and coronavirus (2/202). InterpretationClinical testing methods for 2019-nCoV require improvement. Importantly, 5.8% of 2019-nCoV infected and 18.4% of non-2019-nCoV-infected patients had other pathogen infections. It is important to treat combined infections and perform rapid screening to avoid cross-contamination of patients. A test that quickly and simultaneously screens as many pathogens as possible is needed. FundingNo founding was received Research in contextO_ST_ABSEvidence before this studyC_ST_ABSWe searched PubMed for articles published up to January 31, 2020 using the keywords "2019 novel coronavirus" or "2019-nCoV". No published study on the characteristics of 2019-nCoV tests or 2019-nCoV co-infections was found. We only noted recent laboratory findings for other tests of patients infected with 2019-nCoV. Added value of this studyPositive detection of nCoV-NP or nCovORF1ab is presented, and individuals with/without 2019-nCoV infections or with inconclusive results were identified. Patients with inconclusive results may be diagnosed with 2019-nCoV infection or found to be negative for the infection after resampling and retesting in the next few days. Approximately 5.8% of the subjects diagnosed with 2019-nCoV had co-infection. Implications of all the available evidenceManagement of the population showing inconclusive results should be given attention; additionally, such results can be minimized by improving the sampling, sample pretreatment, and testing methodologies. When diagnosing 2019-nCoV subjects, the possibility of co-infection should be considered. Finally, better clinical detection methods are needed to simultaneously screen as many pathogens as possible.
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BackgroundThe emerging 2019 novel coronavirus (2019-nCoV) has pushed several countries into state of emergency all over the world. The possible transmission of 2019-nCoV by conjunctiva is controversial and has substantial public health implications. MethodsA retrospective cohort study was initiated to investigate the possible transmission of 2019-nCoV through aerosol contact with conjunctiva. We enrolled 67 cases of confirmed or suspected cases of novel coronavirus pneumonia (NCP) during 17-28 Jan 2020. Nasopharyngeal and conjunctival swabs were collected for real time RT-PCR analysis to detect 2019-nCoV. Results63 patients were identified as laboratory-confirmed NCP and the remaining four were suspected NCP. Conjunctival swab samples from one NCP patient yielded positive PCR results and two NCP patients yielded probable positive PCR results. None of the three patients had ocular symptoms. The only one NCP patient with conjunctivitis as the first symptom had negative conjunctival sac 2019-nCoV test. Conjunctival swab samples from the four suspected cases of NCIP were negative. Conclusion2019-nCoV can be detected in the conjunctival sac of patients with NCP. Through clinical analysis, viral transmission via the conjunctival route was not supported by the data. Good clinical protection can effectively cut off the transmission path.
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In December, the outbreak of a novel coronavirus (2019-nCoV) in Wuhan, China, has attracted extensive global attention. On January 20, 2020,the Chinese health authorities upgraded the coronavirus to a Class B infectious disease in the Law of the People's Republic of China on the Prevention and Treatment of Infectious Diseases, and considered it as Class A infectious diseases in disease control and prevention. On January 22, 2020, the 2019-nCoV nucleic acid detection test was listed as the diagnostic criteria in the "guidelines for diagnosis and treatment of pneumonia due to 2019-nCoV (Trial Version 2)" . Therefore, standardizing the operation process of the 2019-nCoV nucleic acid detection in clinical laboratories has become a top priority. It is of paramount importance to establish standard protocols for detection of the 2019-nCoV nucleic acids in clinical laboratories to improve the reliability of the results and ensure the biosafety of laboratory personnel.
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Objective@#To investigate the positive rate for 2019-nCoV tests and co-infections in Wuhan district.@*Methods@#A total of 8 274 cases in Wuhan were enrolled in this cross-sectional study during January 20 to February 9, 2020, and were tested for 2019-nCoV using fluorescence quantitative PCR. Both respiratory tract samples (nasopharynx, oropharynx, sputum and alveolar lavage fluid) and non-respiratory tract samples (urine, feces, anal swabs, blood and conjunctival sac swabs) were collected. If both orf1ab and N genes are positive, they are classified as nucleic acid test positive group; if both orf1ab and N genes are negative, they are classified as negative group; if single gene target is positive, they are classified as suspicious group. Individuals were divided into male group and female group according to sex. At the same time, 316 patients were tested for 13 respiratory pathogens by multiplex PCR.@*Results@#Among the 8 274 subjects, 2 745 (33.2%) were 2019-nCoV infected; 5 277 (63.8%) subjects showed negative results in the 2019-nCoV nucleic acid test; and 252 cases (3.05%) was not definitive (inconclusive result). The age of cases with COVID-19 patients and inconclusive cases was significantly higher than that of cases without 2019-nCoV infection (40 vs 56, t=27.569, P<0.001; 52 vs 56, t=6.774, P<0.001). The positive rate of 13 respiratory pathogens multiple tests was significantly lower in 104 subjects who were positive for 2019-nCoV compared with those in subjects who were negative for 2019-nCoV test (5.77% vs 18.39%, χ2=24.105, P=0.003). Four types of respiratory tract samples and five types of non-respiratory tract samples were found to be positive for 2019-nCoV nucleic acid test.@*Conclusion@#The 2019-nCoV nucleic acid positive rate in male is higher than in female. Co-infections should be pay close attention in COVID-19 patients. 2019-nCoV nucleic acid can be detected in non-respiratory tract samples.
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Objective:To investigate the expression of inflammatory factors, advanced oxidation protein products (AOPP), homocysteine (Hcy), vascular cell adhesion molecule 1 (VCAM-1) and the clinical significance of pregnancy outcome in patients with gestational diabetes.Methods:From January 2019 to January 2020, 103 cases of gestational diabetes in Taizhou Hospital of Zhejiang province were selected as observation group, and 87 normal pregnant women in Taizhou Hospital of Zhejiang province from January 2019 to January 2020 were selected as control group.The fasting blood glucose (FPG) and fasting insulin (FINS) were measured by automatic biochemical analyzer.The levels of tumor necrosis factor-α(TNF-α), high-sensitivity C-reactive protein (hs-CRP), interleukin-6 (IL-6), AOPP and VCAM-1 were measured by enzyme-linked immunosorbent assay (ELISA). The Hcy level was measured by circulating enzyme method.Results:The FPG [(8.34±1.25)mmol/L] and FINS [(9.75±0.89)U/L] in the observation group were higher than those in the control group [(4.89±0.67)mmol/L and (5.93±0.45)U/L] ( t=23.088, 36.297, all P<0.05). The serum levels of TNF-α [(21.63±3.25)ng/L], hs-CRP [(6.34±1.29)mg/L], IL-6 [(147.32±28.39)ng/L] in the observation group were higher than those in the control group [(8.72±1.84)ng/L, (2.17±0.73)mg/L and (87.96±12.41)ng/L] ( t=32.861, 26.744, 18.091, all P<0.05). The serum levels of AOPP [(53.21±9.89)μg/L], Hcy [(11.23±1.36)μmol/L] and VCAM-1 [(94.27±15.46)μg/L] in the observation group were higher than those in the control group [(25.48±6.18)μg/L, (8.41±1.28)μmol/L and (62.18±6.52)μg/L] ( t=22.674, 14.627, 18.047, all P<0.05). The incidence of adverse pregnancy outcome in the observation group (29.13%) was lower than that in the control group (4.60%) (χ 2=19.313, P<0.05). Conclusion:The patients with gestational diabetes have obvious inflammatory reaction, higher serum levels of AOPP, Hcy and VCAM-1, and poor pregnancy outcome.
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In December, the outbreak of a novel coronavirus (2019-nCoV) in Wuhan, China, has attracted extensive global attention. On January 20, 2020, the Chinese health authorities upgraded the coronavirus to a Class B infectious disease in the Law of the People′s Republic of China on the Prevention and Treatment of Infectious Diseases, and considered it as Class A infectious diseases in disease control and prevention. On January 18, 2020, the 2019-nCoV nucleic acid detection test was listed as the diagnostic criteria in the "guidelines for diagnosis and treatment of pneumonia due to 2019-nCoV (Trial Version 2)" . Therefore, standardizing the operation process of the 2019-nCoV nucleic acid detection in clinical laboratories has become a top priority. It is of paramount importance to establish standard protocols for detection of the 2019-nCoV nucleic acids in clinical laboratories to improve the reliability of the results and ensure the biosafety of laboratory personnel.
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Objective:To investigate the positive rate for 2019-nCoV tests and co-infections in Wuhan district.Methods:A total of 8 274 cases in Wuhan were enrolled in this cross-sectional study during January 20 to February 9 in 2020, and were tested for 2019-nCoV using fluorescence quantitative PCR. Both respiratory tract samples (nasopharynx, oropharynx, sputum and alveolar lavage fluid) and non-respiratory tract samples (urine, feces, anal swabs, blood and conjunctival sac swabs) were collected. If both orf1ab and N genes are positive, they are classified as nucleic acid test positive group; if both orf1ab and N genes are negative, they are classified as negative group; if single gene target is positive, they are classified as suspicious group. Individuals were divided into male group and female group according to sex. At the same time, 316 patients were tested for 13 respiratory pathogens by multiplex PCR.Results:Among the 8 274 subjects, 2 745 (33.17%) were 2019-nCoV infected; 5 277 (63.77%) subjects showed negative results in the 2019-nCoV nucleic acid test; and 252 cases (3.05%) was not definitive (inconclusive result). The age of cases with COVID-19 patients and inconclusive cases was significantly higher than that of cases without 2019-nCoV infection (56>40, t=27.569, P<0.001; 52>40, t=6.774, P<0.001). The positive rate of 13 respiratory pathogens multiple tests was significantly lower in 104 subjects who were positive for 2019-nCoV compared with those in subjects who were negative for 2019-nCoV test (5.77% vs 18.39%, χ 2=24.105, P=0.003). Four types of respiratory tract samples and five types of non-respiratory tract sampleswere found to be positive for 2019-nCoV nucleic acid test. Conclusion:The 2019-nCoV nucleic acid positive rate inmale is higher than infemale. Co-infections should be pay close attention in COVID-19 patients. 2019-nCoV nucleic acid can be detected in non-respiratory tract samples.
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Acute respiratory tract infections ranks first in China for various infectious diseases.Lower respiratory tract infections and related diseases caused a heavy burden on China′s medical care and society. In particular, COVID has caused great losses. This article discusses the standardization of clinical pathological diagnosis of respiratory pathogen infection, in order to improve the correct diagnosis of the disease and facilitate the timely treatment of the disease.
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High-throughput sequencing, also known as next-generation sequencing (NGS), is a new type of genetic screening and diagnostic technology. Its continuous innovation acceleratesthe understanding of genetic markers and the molecular mechanisms of disease, especially for complex hereditary diseases. With the development of NGS technology, especially the emergence of the targeted genome sequencing, genome-wide exome sequencing and genome-wide sequencing projects, NGS has been clinically applied indetection of SNVs, structural rearrangements and CNVs, monitoring circulating tumor DNA, and analyzing the genome regions that previously challenged the management of standard bioinformatics algorithms.
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Objective To establish the system of high resolution melting for detection of aldehyde dehydro-genase 2(ALDH2) and alcohol dehydrogenase-1B (ADH1B) gene polymorphisms .Methods The short prim-ers were designed for ALDH2 and ADH1B gene .Different PCR products were analyzed using Eva Green dyes after amplification ,which were confirmed by Sanger sequencing .Results The genotypes of ALDH2 rs671 and ADH1B rs1229984 were successfully detected in the same procedure by HRM within 90min ,and the results were consistent with Sanger sequencing .Conclusion The assay of HRM is simple ,rapid ,cost-effective ,and re-liable for the detection of ALDH2 and ADH1B polymorphism and it is worthy to be popularized .
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Objective To observe the effect of Yunnanbaiyao combined with gelatin sponge on wound healing in patients with post-extraction hemorrhage.Methods 72 patients received dental extraction were selected,and they were divided into observation group (Yunnanbaiyao + gelatin sponge) and control group (gelatin) according to the digital table,each group in 36cases.The hemostasis after treatment and the improvement of pain after 12h and 24h after treatment were observed.The wound healing was observed in the two groups after 7 days of treatment.Results After treatment,the total effective rate was 91.67％ in the observation group and 69.44％ in the control group.The effective rate of hemostasis in the observation group was significantly higher than that of the control group(x2 =7.32,P =0.007).The VAS scores of the observation group at 12 h (t =23.44,P =0.000) and 24h (t =22.86,P =0.000) after pack treatment obviously decreased;The VAS scores of the control group at 12h(t =19.87,P =0.000) and 24h (t =18.47,P =0.000) after pack treatment obviously decreased;The VAS scores of the observation group at 12h and 24h after pack treatment was lower than those of the control group,the differences were statistically significant(t =7.03,5.03,all P =0.000).The wound healing of the observation group was improved after 7d treatment(t =8.12,P =0.00);The wound healing of the control group was improved after 7d treatment(t =5.39,P =0.00);The wound healing of the observation group was better than that of the control group,the differences were statistically significant (t =2.88,P =0.005).Conclusion Yunnanbaiyao combined with gelatin sponge can relieve the bleeding symptoms after tooth extraction,promote wound healing,reduce the patients' pain.
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Objective To investigate the association of rs10004195 single nucleotide polymorphisms in Toll like re-ceptor 10(TLR 10) gene with helicobacter pylori infection and associated diseases. Methods A total of 652 pa-tients who has been examined by gastroscopy were obtained, and then peripheral blood samples were collected from all the patients. Immune colloidal gold method was used to test the serological Hp antibody of all the patients. The TLR10 gene rs10004195 polymorphisms were examined by direct DNA sequencing of the PCR products. Results The frequencies of AA,TT and AT genotype on TLR10 rs10004195 were 30. 98%, 20. 71%, 48. 31%;there was significant difference between Hp antibody positive group and Hp antibody negative group in the TT frequencies of TLR10 rs10004195 ( P<0. 05 ) . No significant difference between controls and Hp associated diseases groups in Hp antibody positive group or in Hp antibody negative group were observed. Conclusion There was correlation be-tween the TLR10 rs10004195 loci genotype and the risk of Hp infection, but no correlation with Hp associated dis-eases.
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Objective To evaluated the prevalence of EGFR mutations in Hubei region,to provide reliable experimental basis for reasonable screening TKI beneficiaries in clinic.Methods EGFR mutation of 253 patients diagnosed with NSCLC in Hu-bei region during 2010 to 2015 was detected by PCR-sanger sequencing and ARMS assay,to investigate the sensitivity of dif-ferent methods,to explore the frequency and clinical significance of EGFR mutation in different gender,in different histologi-cal type and different exons.Results 93 of 253 NSCLC patients harbored EGFR mutation,mainly occurred in exon 19 and 21,accounting for 53.76% and 35.38% of the total EGFR mutation rate,respectively.ADx-ARMS method showed higher sensitivity (P=0.001).The mutation detection rate of female NSCLC was significantly higher than that of male NSCLC (P=0.001).The observed incidence of EGFR mutations in patients with adenocarcinomas (38.01%)was the highest in differ-ent histological type,followed by glandular squamous cell carcinoma (30.77%),large cell carcinoma (20%)and squamous cell carcinoma (4.55%).Nonsmoking patients had a higher EGFR mutation frequency (51.6%,81/157)than those with a history of smoking (24%,12/50).Conclusion ARMS assay was more sensitive and more convenient detection method for clinical screening for EGFR-TKI treatment subpopulation.The mutation rate of female NSCLC was significantly higher than that of male NSCLC in Hubei Province.Speculated that the sexual differences in NSCLC with EGFR mutation frequencies were related to hormone levels and smoking status.
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With the development of molecular diagnostic techniques, the study for the etiology of leukemia has been entering the era of the molecular biology.Molecular techniques for leukemia diagnosis, prognosis and individualized therapy are used widely, becoming one of the necessary routine tests for patients with leukemia. So far, molecular techniques for leukemiaincluding cytogenetic diagnosis, molecular genetics, molecular diagnostics based on PCR, mutation detection, as well as a variety of next-generation sequencing, have played an important role in the diagnosis of leukemia, minimal residual disease monitoring, prognosis and targeted therapy.
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Objective Investigating the expression of TLR3 and TLR7 mRNA in peripheral blood mononuclear cells (PBMC) of patients with chronic hepatitis C infection (CHC),to explore the effects of gender and age on Toll-like receptor expres-sion.Methods Peripheral blood was collected from 1 1 5 patients of chronic hepatitis C infection,1 1 3 healthy individuals.Ex-pression levels of TLR3 and TLR7 mRNA were detected by real-time quantitative PCR.Results The expression of TLR3 and TLR7 mRNA were significant difference between patients with CHC infection and healthy individuals (t=38.73,6.16, P0.05).The HCV-RNA load of premenopausal females was significant lower than young males and old females (t=3.49,2.51,P<0.05),the load of old males was lower than old females (t=2.35,P<0.05),however, there was no significant differences between old males and old females (t=1.20,P<0.05).Conclusion Gender and age dis-crepancies have a relationship with the expression of Toll-like receptors of patients with CHC infection,which may provide a theoretical basis and a new method for CHC.