1.Application of nucleic acid detection and antibody detection in HIV infection
Haiyan HOU ; Qingli YAN ; Jing LIU ; Chuncheng LIU
International Journal of Laboratory Medicine 2015;(23):3431-3432
Objective To compare the accuracy of nucleic acid detection and antibody detection for diagnosing human immunode‐ficiency virus(HIV) infection .Methods Retrospectively analysed data of nucleic acid detection and antibody detection from 124 ca‐ses of patients diagnosed with HIV infection from 2005 to 2014 .The positive rates of the two methods were compared respectively in patients with early‐stage of HIV infection(76 cases) and patients with intermediate and advanced stage of HIV infection (48 ca‐ses) .Results In patients with early‐stage of HIV infectionn ,the positive rate of nucleic acid detection (94 .74% ) was higher than that of antibody detection (84 .21% );while in patients with intermediate and advanced stage of HIV infection ,the positive rate of antibody detection(97 .92% ) was higher than that of nucleic acid detection (81 .25% );both had statistically significant difference (P<0 .05) .Conclusion On the early stage of HIV infection ,the accuracy of nucleic acid detection is higher than that of antibody detection ;while on the intermediate and advanced stage of HIV infection ,antibody detection shows better accuracy .
2.Establishment of TaqManˉBHQ probe realˉtime fluorescence RTˉPCR for quantitative detection of Soul virus
Chuncheng LIU ; Pengfei YANG ; Qingli YAN ; Yujun SHI
International Journal of Laboratory Medicine 2014;(24):3332-3333
Objective To establish a rapid method of real-time fluorescence RT-PCR to quantify Soul virus.Methods The pro-fessional software was adopted to design the primer and the TaqMan-BHQ probe.With artificially synthesized L gene segment as the template of Soul virus,the real-time RT-PCR for detecting Soul virus was researched.Results The Ct value of templates had a good linear relationship with the log value of the template diluted concentration.The standard curve was Y =-3.607X +41.84, r2 =0.998,the PCR amplification efficiency was 108.1%,its lowest detection limit was 53.2 copies/μL.Conclusion Applying the real-time fluorescence RT-PCR by the TaqMan-BHQ probe for detecting nucleic acid of Seoul virus has the characteristics of short time-consuming and high sensitivity.
3.The epidemiologic and virological analysis of an outbreak of influenza A/H1N1
Haiyan HOU ; Pengfei YANG ; Minhui ZHANG ; Jing LIU ; Chuncheng LIU ; Yujun SHI
Chinese Journal of Experimental and Clinical Virology 2015;29(5):405-408
Objective To study the epidemiological characters of an influenza A/H1N1 outbreak in a middle school in Huaian city,and to provide scientific measures for prevention and control of influenza A/ H1N1.Method The outbreak of influenza A/H1N1 was investigated through the field epidemiology.The samples were used for virus isolation.Nucleotide sequences of H1N1 HA1 region were determined by RT-nested-PCR and sequencing.H1N1 genotypes were analyzed using bioinformatic programs by MEGA 5.0 software,and phylogenetic tree was constructed.Result 51 cases were reported during 11 days,the attack rate was 2.49% (51/2051).The sequence analysis showed that the sequence homologous rates of H1N1 virus HA gene nucleotide were 99.6%-99.9% with H1N1 genotype in 2014,98.5%-98.8% in 2013,and lower than 98.4% in 2011.Conclusion The popularity of this outbreak was low.Timely isolation of H1 N1 patients and close contacts played an important role in control the epidemics.Implementation of health propaganda and immunization of influenza A/H1N1 should be strengthened.
4.Synthesis and Identification of Hapten and Complete Antigen of Norketa mine
Tingting YANG ; Chuncheng ZHAO ; Xiaoping SHEN ; Bochang TAN ; Pankun ZOU ; Huaiming WANG ; Lingyun LIU ; Chao WEI ; Wanli WEI ; Jinhai HE ; Jie WU ; Dongsheng ZHANG ; Xiaolian ZHAO
Chinese Journal of Analytical Chemistry 2010;38(1):109-112
Under low temperature conditions, the hapten carboxyl-norketamine was synthesized by reacting norketamine and succinaldehyde acid.Identification result using electrospray ionization mass showed the hap ten was successfully synthesized.The artificial antigen confirmed by infrared spectroscopy was developed by conjugating hapten to carrier proteins with carbodiimide(EDC) method.Matrix-assisted laser desorption ioni zation time of flight mass spectrometry(MALDI-TOF-MS) showed that the ratio of hapten to BSA was 11:1.The antibody with high titer(5.12 × 10~4) was produced after immuning to rabbits.
5.Fatty liver and light manual labor: an epidemiological survey in Tianjin by controlled attenuation parameter
Hai LI ; Chuncheng LIU ; Yulin WANG
Journal of Clinical Hepatology 2015;31(7):1073-1077
ObjectiveTo investigate the incidence and possible related factors for fatty liver among workers engaged in light manual labor in Tianjin, China. MethodsAmong 800 healthy workers engaged in light manual labor randomly selected from a company in Tianjin, a total of 602 healthy workers completed physical examination and data collection. The content of fat and liver stiffness were determined by controlled attenuation parameter (CAP) using FibroScan, and the disease history, body mass, height, blood pressure, and living habits in all subjects were investigated. The possible related factors for different degrees of fatty liver were analyzed. Between-group comparison of continuous data was performed using the Kruskal-Wallis H test, and comparison of categorical data was performed using χ2 test with row×column tables. Pairwise comparison was performed using χ2 segmentation method, and the agreement between CAP and color ultrasound was analyzed using the Kappa coefficient. ResultsAmong the 602 subjects, there were 180 (29.9%), 195 (32.3%), and 227 (37.7%) subjects with severe fatty liver, moderate fatty liver, and no fatty liver, respectively. There were significant differences in age, body mass index, waist-hip ratio, smoking, drinking, history of diabetes, and family history between the above three groups according to CAP (P<005 for all). With higher sensitivity, CAP achieved a significantly higher detection rate of fatty liver than traditional color ultrasound (623% vs 44.0%, P=0.000). The agreement between two methods was poor (Kappa value <0.4). ConclusionAmong the so-called “healthy people” engaged in light manual labor, the incidence of fatty liver has exceeded 60%, which is associated with body mass index, waist-hip ratio, bad living habits, history of diabetes, and family history of fatty liver. CAP can noninvasively detect fatty liver with higher sensitivity and less time; however, its clinical significance needs further studies.
6.The therapeutic evaluation of video-assisted anal fistula treatment for Parks Ⅱ anal fistula
Yuru ZHANG ; Xiu ZHANG ; Bin HUANG ; Liancheng LIU ; Chuncheng REN ; Min WANG ; Tuanjie ZHAO ; Zhiliang ZHANG ; Xu HAN
Chinese Journal of General Surgery 2018;33(8):658-661
Objective To evaluate video-assisted anal fistula treatment (VAAFT) for Parks type Ⅱ anal fistula.Methods 40 Parks type Ⅱ anal fistula patients underwent VAAFT procedure from June 2015 to June 2017.Results were compared with 40 cases treated by incision and thread drawing.Results There was no significant difference between the two groups for curative effect,postoperative urinary retention,wound edema,bleeding and recurrence rate after 6 months of operation (90% vs.95%,x2 =0.722,P =0.697;5% vs.8%,x2 =0.213,P=1.0;2% vs.8%,x2 =1.053,P=0.615;0 vs.5%,x2 =2.051,P =0.494;10% vs.5%,x2 =0.721,P =0.675).Pain on first day and one week after operation in the VAAFT was less [(1.9±0.6) vs.(3.7±1.0),t =9.438,P=0.001;(0.9±0.7) vs.(1.9±0.8),t=6.269,P=0.001],hospital stay was shorter [8.4 ±1.3) d vs.(9.2 ±2.2) d,t =2.030,P=0.047],wound healing was faster [(27 ±8) dm.(38 ±6) d,t =7.328,P =0.001].The Jorge-Wexner incontinence score [(0.5±0.7) vs.(1.2±1.3),t =2.951,P=0.005] and the fecal incontinence severity index [(1.1±1.6) vs.(5.1 ±3.2),t =7.097,P=0.001] were lower in patients receiving VAAFT procedure.Conclusion Video-assisted anal fistula treatment is a safe and effective surgical method with the advantages of less trauma,and pain,quicker recovery and no damage to the anal sphincter.
7.Modified Bascom cleft lift procedure for chronic pilonidal sinus
Liancheng LIU ; Yuru ZHANG ; Hongyan DUAN ; Yue SU ; Bin HUANG ; Xu HAN ; Chuncheng REN ; Min WANG ; Tuanjie ZHAO ; Zhiliang ZHANG ; Fang XIONG
Chinese Journal of General Surgery 2019;34(1):42-44
Objective To evaluate the clinical efficacy of modified Bascom cleft lift procedure in the treatment of chronic sinus.Methods Modified Bascom cleft lift procedure was performed in 53 patients admitted from Oct 2012 to Jul 2016.49 cases were male and 4 were female.The average age was (25.4± 2.3) years.Results All patients were satisfied with the operation.There were 49 cases of primary healing and 4 cases of incision complications.The average follow-up was (12.1 ±4.3) months,no recurrence was observed.Conclusion The modified Bascom cleft lift technique is effective and reliable,with less complications and a lower recurrence rate.
8.Analysis of the role of endoscopic retrograde cholangiopancreatography in children with pancreaticobiliary diseases.
Wei LIU ; Qiming WANG ; Hongze ZENG ; Yi MOU ; Hang YI ; Chuncheng WU ; Bing HU ; Chengwei TANG
Chinese Journal of Pediatrics 2014;52(5):328-332
OBJECTIVETo evaluate the safety and efficacy of endoscopic retrograde cholangiopancreatography (ERCP) in children with pancreaticobiliary diseases and the characteristics of pancreaticobiliary disorders in children.
METHODRetrospective review was conducted on the data of patients younger than 18 years who underwent ERCP between 2005 and 2012 at West China Hospital. The indications,ERCP findings, ERCP procedures, complications, and clinical outcomes were evaluated.ERCP procedures were performed using standard duodenoscopes under general anaesthesia or sedation, which included all endoscopic treatments, such as endoscopic sphincteropapillotomy, stone extraction, stent treatment and so on.
RESULTOne hundred and two ERCPs were performed on 68 patients, and all the procedures were successfully completed in 100% cases. There were 39 girls (57%), and median age at time of procedure was 14.6 years (range, 5-17 years).General anesthesia and sedation were performed in 81% and 19% of procedures, respectively. The ERCP findings were classified as follows:bile duct stone(s) (n = 37, 54%), pancreatic duct stone(s) (n = 8, 12%), bile duct benign stricture (n = 7, 10%) and other nonmalignant pancreaticobiliary diseases (n = 16, 24%).Four cases (4/102, prevalence 4%) were complicated with post-ERCP pancreatitis.Symptoms such as abdominal pain and jaundice were cured obviously after the procedures of ERCP were performed.
CONCLUSIONThe main characteristics of pancreaticobiliary disorders in children were nonmalignant pancreaticobiliary diseases, such as bile duct stone, pancreatic duct stone, and bile/pancreatic duct benign stricture.When performed by well-trained endoscopists, ERCP is safe and effective in children.
Adolescent ; Biliary Tract Diseases ; diagnostic imaging ; surgery ; Calculi ; diagnosis ; pathology ; surgery ; Child ; Child, Preschool ; Cholangiopancreatography, Endoscopic Retrograde ; Choledocholithiasis ; diagnosis ; pathology ; surgery ; Female ; Humans ; Male ; Pancreatic Diseases ; diagnosis ; pathology ; surgery ; Pancreatic Ducts ; diagnostic imaging ; surgery ; Retrospective Studies ; Treatment Outcome
9.Inhibiting effect of miR-18a on expression of extracellular matrix-related genes of NIH-3T3 by targeting Notch2
Jiaqi HAO ; Jing WEN ; Chuncheng LIU ; Lu CAI
Journal of Environmental and Occupational Medicine 2022;39(7):786-791
Background The main pathological feature of silicosis is pulmonary fibrosis. Multiple miRNAs regulate the development of silicosis. Objective Using a fibroblast cell line, to explore the effect of miR-18a on the expression of extracellular matrix-related genes, and verify the mechanism. Methods The fibroblast cell line NIH-3T3 cells were transfected with miR-18a mimics or neurogenic locus notch homolog protein 2 (Notch2) small interfering RNA (siRNA). The mRNA expression changes of Acta2, Col1a1, and Notch2 were detected by real-time quantitative reverse transcription PCR (qRT-PCR), α-smooth muscle actin (α-SMA) and Notch2 were also detected at the protein level by Western blotting. To verify whether miR-18a could directly act on the complementary sequences of the Notch2 gene, human embryonic kidney HEK293T cells and the psiCHECKTM-2 vector were used. Results The results of qRT-PCR showed that in NIH-3T3 cells, the over-expression of miR-18a mimics for 36 h inhibited the mRNA expression of Col1a1 and Acta2 (P<0.05). The results of Western blotting showed that the protein expression abundance of α-SMA was decreased at 48 h of miR-18a mimics over-expression. The qRT-PCR results showed that the over-expression of miR-18a for 36 h had no significant effect on Notch2 gene expression, but the Western blotting results showed that the over-expression of miR-18a mimics inhibited the expression of Notch2 at the protein level. The results of the dual luciferase reporter vector assay showed that in HEK293T cells, both over-expressed miR-18a mimics and inhibitors for 24 h demonstrated that Notch2 is a direct target gene of miR-18a. When Notch2 was inhibited for 36 h, the qRT-PCR results showed that Acta2 and Col1a1 were down-regulated (P < 0.05), and the results of Western blotting showed that α-SMA protein was also inhibited. Conclusion The findings indicate that miR-18a could inhibit the expression of extracellular matrix-related genes of NIH-3T3 cells by directly acting on the 3’UTR of target gene Notch2.
10. Analysis of HA1 gene of influenza A H1N1 pdm09 virus from a clustered human cases
Pengfei YANG ; Qingli YAN ; Xuezheng MA ; Liping ZHANG ; Wei ZHEN ; Chuncheng LIU ; Yadong XING ; Haibo YAO ; Nanjiang HE ; Kongxin HU
Chinese Journal of Experimental and Clinical Virology 2017;31(1):42-46
Objective:
To understand the viral etiology of a clustered case of human infection outbreak in the middle school of Huai’an city.
Methods:
Nasopharyngeal swab samples from patients were collected and rapidly detected by Real-time RT-PCR and the target virus isolated in cells. Furthermore, HA1 segments of target virus were amplified by RT-PCR and sequenced. The genetic and phylogenetic analysis based on HA1 genes was computed.
Results:
Influenza A(H1N1)pdm09 viral nucleic acid in 11 nasopharyngeal swab samples from patients in the outbreak were positive. Compared to the vaccine strains A/California/07/2009, the Huai’an isolates, nucleotide identity was 97.7%-98.1%, and amino acid identity was 96.6%-97.4%. Phylogenetic analysis of HA1 segment sequences indicated that the Huai’an strains from the outbreak were related closely to the viruses isolated in the year of 2014. Sequence analysis indicated that the Huai’an isolates had no amino acid substitution in the receptor binding sites and glycosylation sites, while in the Ca1 of antigenic determinant of HA1 the Huai’an isolates had an amino acid substitution of S for T at 220.
Conclusions
The pathogen of the clustered case of human infection was Influenza A(H1N1)pdm09 virus. Though the Huai’an isolates had one animo acid substitution in the Ca1 of antigenic determinant, the antigenicity characteristic remained unchanged.