1.Preparation of quality control materials for SARS-CoV-2 variants based on MS2 phage virus-like particles
Ran ZHAO ; Yingwei CHEN ; Chengxiang CHU ; Zhongqiang HUANG ; Weijie DING ; Xueliang WANG
Chinese Journal of Clinical Laboratory Science 2025;43(10):773-779
Objective To prepare a variety of quality control(QC)materials for SARS-CoV-2 variants as an addition to the conven-tional SARS-CoV-2 nucleic acid QC products for the laboratory detection of mutant strains by optimizing the preparation and purification process of MS2 phage virus-like particle(VLP)technique,and evaluate their performances.Methods The typical mutation sequence fragments or full length S genes were designed and synthesized according to the genomic information of SARS-CoV-2 variants.Then,they were inserted into the downstream of maturase gene,coat protein and the pac-site of MS2 phage to construct a series of recombi-nant expression vectors.After induced by the prokaryotic expression system,the VLP products were purified through the polyethylenei-mine precipitation,ultrafiltration,nuclease digestion,and gel filtration chromatography.The obtained VLP were validated by the nucle-ic acid electrophoresis,protein electrophoresis,protein concentration determination,and fluorescence PCR,and their performances such as nucleic acid residue and stability were also evaluated.Results A total of 10 kinds of VLP containing the targeted sequences of the gene to be tested were prepared.The length of the foreign sequence wrapped in them ranged from 297 bp to 3 822 bp,which could be combined into a variety of QC materials for the mutation detection of different SARS-CoV-2 variants.The prepared VLP QC materials could not be effectively amplified without nucleic acid extraction or reverse transcription steps during the routine nucleic acid detection.The simulated QC samples remained stable after repeated freeze-thaw cycles.They could be stored stably for 2 months at 25 ℃ and 4 weeks at 37 ℃.Conclusion The established preparation and combined purification process of VLP QC materials can encapsulate vari-ous exogenous nucleic acid sequences with different lengths into the viral coat protein to form VLP,with high production efficiency.The VLP QC products prepared by the above process have stable performance and almost no residual exogenous nucleic acid,which can ef-fectively meet clinical requirements and ensure the quality of laboratory testing.
2.Preparation of quality control materials for SARS-CoV-2 variants based on MS2 phage virus-like particles
Ran ZHAO ; Yingwei CHEN ; Chengxiang CHU ; Zhongqiang HUANG ; Weijie DING ; Xueliang WANG
Chinese Journal of Clinical Laboratory Science 2025;43(10):773-779
Objective To prepare a variety of quality control(QC)materials for SARS-CoV-2 variants as an addition to the conven-tional SARS-CoV-2 nucleic acid QC products for the laboratory detection of mutant strains by optimizing the preparation and purification process of MS2 phage virus-like particle(VLP)technique,and evaluate their performances.Methods The typical mutation sequence fragments or full length S genes were designed and synthesized according to the genomic information of SARS-CoV-2 variants.Then,they were inserted into the downstream of maturase gene,coat protein and the pac-site of MS2 phage to construct a series of recombi-nant expression vectors.After induced by the prokaryotic expression system,the VLP products were purified through the polyethylenei-mine precipitation,ultrafiltration,nuclease digestion,and gel filtration chromatography.The obtained VLP were validated by the nucle-ic acid electrophoresis,protein electrophoresis,protein concentration determination,and fluorescence PCR,and their performances such as nucleic acid residue and stability were also evaluated.Results A total of 10 kinds of VLP containing the targeted sequences of the gene to be tested were prepared.The length of the foreign sequence wrapped in them ranged from 297 bp to 3 822 bp,which could be combined into a variety of QC materials for the mutation detection of different SARS-CoV-2 variants.The prepared VLP QC materials could not be effectively amplified without nucleic acid extraction or reverse transcription steps during the routine nucleic acid detection.The simulated QC samples remained stable after repeated freeze-thaw cycles.They could be stored stably for 2 months at 25 ℃ and 4 weeks at 37 ℃.Conclusion The established preparation and combined purification process of VLP QC materials can encapsulate vari-ous exogenous nucleic acid sequences with different lengths into the viral coat protein to form VLP,with high production efficiency.The VLP QC products prepared by the above process have stable performance and almost no residual exogenous nucleic acid,which can ef-fectively meet clinical requirements and ensure the quality of laboratory testing.
3.Clinical study on the imbalance of gut microbiota mediated by foodborne antibiotic exposure leading to irritable bowel syndrome
Qiulan HUANG ; Min LU ; Nali XUE ; Xinqi WANG ; Shouxia GAO ; Hongchou LIU ; Yingwei MA
International Journal of Laboratory Medicine 2025;46(19):2347-2352
Objective To investigate the relationship between exposure to foodborne antibiotics and gut microbiota,as well as its impact on irritable bowel syndrome(IBS).Methods A total of 100 IBS patients(IBS group)and 100 healthy controls(control group)were enrolled in this case-control study.Antibiotic ex-posure was assessed by serum detection,and gut microbiota composition was analyzed by 16S rRNA gene se-quencing technology.Binary Logistic regression model and chi-square test were used for statistical analysis to explore the relationship between antibiotic exposure and IBS,and to analyze the differences in gut microbiota.Results The antibiotic exposure rate of IBS group was significantly higher than that of control group(59%vs.36%,P=0.016),and the exposure to β-lactam antibiotics was significantly different between the two groups(P=0.047).In addition,the composition of gut microbiota in the IBS group was significantly different from that in the control group,and the abundance of Firmicutes was increased(P<0.001),while the abun-dance of Lactobacillus,Bifidobacterium and Faecalibacterium was decreased(P<0.001).Conclusion Food-borne antibiotic exposure may be one of the risk factors for IBS,and antibiotic exposure may affect the occur-rence and development of IBS by changing the composition of gut microbiota.Foodborne antibiotic exposure may be a risk factor for inducing IBS and could affect the disease process by altering the structure of the gut microbiota.Therefore,enhancing the regulation of foodborne antibiotic use and raising public awareness about the importance of minimizing foodborne antibiotic exposure can effectively prevent and reduce the incidence of IBS.
4.A randomized controlled study on the long-term efficacy of intra-cervical lymphatic immunotherapy for adult allergic rhinitis
Kai WANG ; Yang QIN ; Qixing WANG ; Weijun HUANG ; Qingqing YU ; Yin LI ; Yu XIONG ; Yingwei GUO ; Jun TANG
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2023;58(9):871-877
Objective:To determine the long-term efficacy and safety of intra-cervical lymphatic immunotherapy (ICLIT) for adult allergic rhinitis (AR) by comparing it with subcutaneous immunotherapy (SCIT).Methods:A total of 100 adult AR patients with dust mite allergy in Department of Otorhinolaryngology, First People′s Hospital of Foshan from Feb 2018 to Dec 2019 were randomly divided into two groups, 50 in SCIT group [including 42 males and 8 females, aging (32.55±9.72) years] and 50 in ICLIT group [including 45 males and 5 females, aging (31.33±9.84) years]. The changes in total symptom score (total system score, TSS), nasal symptom score (total nasal symptom score, TNSS), eye symptom score (total ocular scoring system, TOSS), drug score (total medication score, TMS), and quality of life score of the two groups of patients were evaluated before and after treatment, and the adverse reactions of all patients during the treatment period were recorded. The changes in the level of dust mite specific IgE (sIgE) in the serum were evaluated. GraphPad Prism 9.0 software was used for statistical analysis.Results:In the SCIT group, 38 patients completed treatment and follow-up, with a dropout rate of 24%. In the ICLIT group, 48 patients completed treatment and follow-up, with a dropout rate of only 4%. The scores of TSS, TNSS, TOSS, TMS, and quality of life in the ICLIT group before treatment were 32.1±3.0, 27.3±3.1, 4.8±2.8, 2.3±0.9, and 68.1±28.7, respectively; After 36 months of treatment, the scores were 21.8±11.4, 18.1±9.4, 3.7±2.9, 1.3±1.1, and 36.0±26.7, respectively, which were significantly lower than those before treatment (all P<0.001). After 36 months of treatment, the TSS of the ICLIT group improved by 10.3±11.2 compared to before, while the TSS of the SCIT group improved significantly by 21.9±11.0 compared to before, with statistically significant differences between the groups ( P<0.001). No serious systemic adverse reactions occurred in both groups of patients. Conclusions:ICLIT treatment for adult AR has long-term efficacy, high safety, and high compliance, but its long-term efficacy is not as good as SCIT. ICLIT can be considered as a new complementary option for AR immunotherapy.
5.The research of pulmonary function changes after thoracoscopic lobectomy versus thoracoscopic segmentectomy based on propensity score matching method
Nadier YIMIN ; Zhouyi LU ; Yunbiao BAI ; Kaiheng GAO ; Yulong TAN ; Xuan WANG ; An WANG ; Dong XU ; Dayu HUANG ; Zhenhua HAO ; Huijun ZHANG ; Ning WU ; Shaohua WANG ; Qinyun MA ; Yingwei WANG ; Xiaofeng CHEN
Chinese Journal of Thoracic and Cardiovascular Surgery 2022;38(1):1-4
Objective:To compare the effects of thoracoscopic anatomical segmentectomy and thoracoscopic lobectomy on patients' respiratory function.Methods:Retrospective analysis of 326 patients who underwent thoracoscopic surgery from July 2016 to July 2019(209 patients underwent anatomical segmentectomy, 117 patients underwent lobectomy). According to variables including gender, age, tumor location, smoking history and BMI, two propensity score-matched cohorts including 89 patients respectively were constructed. The patients’ baseline data and respiratory function date of the patients pre-operation and post-operation were analyzed. The measurement data that obey the normal distribution were described by mean±standard deviation, and the t-test was used for comparison between groups; the measurement data of non-normal distribution was described by the median value( P25, P75), and the Wilcoxon rank sum test was used for the comparison between groups; The data was described by frequency, and the chi-square test or Fisher's exact probability method was used for comparison between groups. Results:At the first-month follow-up after surgery, there was no significant difference in the variation of FVC[(0.48±0.40)L vs.(0.34±0.37)L, P=0.215)and FEV1[(0.52±0.46)L vs.(0.43±0.77)L, P=0.364), and in the change rate of FVC(%)[15.23(8.74, 21.25) vs. 14.58(7.75, 19.40), P=0.122], FEV1(%)[17.25(9.56, 22.78) vs. 16.42(9.15, 20.28), P=0.154]and DLCO(%)[18.54(10.88, 25.68)vs. 17.45(9.58, 23.75) P=0.245]. Between the segmentectomy group and lobectomy group, there was a significant difference in the alteration of FVC[(0.50±0.47)L vs. (0.29±0.31)L, P=0.031] and FEV1[(0.44±0.34)L vs.(0.24±0.23)L, P<0.001], the change rate of FVC(%)[14.27(7.87, 22.32) vs. 9.95(5.56, 17.24), P=0.008]、FEV1(%)[15.23(8.36, 22.17)vs. 10.05(5.15, 18.54), P<0.001]and DLCO(%)[13.74(6.24, 19.78) vs. 4.45(-2.32, 13.75), P=0.023]in the 6th month after surgery. The lobectomy group had a higher variation of FEV1[(0.34±0.49)L vs.(0.18±0.26)L, P=0.006] and change rate of FVC(%)[9.28(2.15, 18.94) vs. 5.24(0.52, 11.45), P=0.0032] and FEV1(%)[10.45(3.15, 21.32) vs. 6.50(1.55, 14.24), P<0.001] in the first year after surgery. However, the variation of FVC[(0.29±0.36)L vs.(0.21±0.24)L, P=0.176) and the change rate of DLCO(%)[8.35(2.15, 16.45) vs. 6.23(2.12, 14.54), P=0.143] didn't show a significant difference between the two groups. Conclusion:Whether in the short or the middle postoperative period, segmentectomy can preserve postoperative respiratory function than lobectomy.
6.Analysis of etiology and clinical characteristics of 618 children with severe community-acquired pneumonia
Yanzhi HUANG ; Liwei SUN ; Yuqi LIU ; Yingwei MA ; Yuling TIAN ; Yanling ZHAO ; Liwu WANG ; Hongbo JIANG ; Aili CUI ; Yan ZHANG ; Wenbo XU
Chinese Pediatric Emergency Medicine 2021;28(2):111-115
Objective:To understand the etiology and clinical characteristics of hospitalized severe community-acquired pneumonia(SCAP) in Changchun, and provide scientific basis for its etiology diagnosis and targeted treatment.Methods:The study subjects included 618 children with clinical diagnosis of SCAP who were hospitalized from January 2016 to December 2019.We collected pharyngeal swabs and alveolar lavage fluid from children.Virus isolation, bacterial culture, time-of-flight mass spectrometry, PCR/RT-PCR, colloidal gold method and Optochin test were used to detect the antigen, nucleic acid and protein profiles in the specimen.Results:There were more boys than girls in hospitalized children with SCAP.The peak age of onset was 7 to 12 months.Most cases occurred in winter and spring.The highest detection rate of SCAP virus was 56.15%(347/618); 73.49%(255/347) were positive for one virus, among which the top five were respiratory syncytial virus (27.8%), influenza A virus (23.9%), influenza B virus (16.1%), rhinovirus (12.2%) and metapneumovirus (10.2%). Two viruses were positive for 19.88%(69/347); three viruses were positive for 4.32%(15/347); four viruses were positive for 2.31%(8/347). Atypical microbial infections were 29.77%(184/618), of which Mycoplasma pneumoniae accounted for 95.65%(176/184). Bacterial infections were 17.31%(107/618), mainly Streptococcus pneumoniae(39.25%, 42/107) and Staphylococcus aureus(24.30%, 26/107). The mixed infection of multiple pathogens was 7.61%(47/618), among which the mixed infection rates of Mycoplasma pneumonia with Streptococcus pneumoniae, virus were 40.43% and 34.04%, respectively.High fever, faster breathing, and perioral cyanosis were risk factors for SCAP, with OR and 95% CI of 7.71 and 4.56-13.04, 2.43 and 2.02-2.93, 3.53 and 2.56-4.86, respectively.Viral co-infection occurred in 36.96%(34/92) of complications such as heart failure, toxic encephalopathy, and myocardial damage; Mycoplasma pneumoniae and other pathogens co-infected 35.29% of children with pleural effusion. Conclusion:The pathogens of SCAP in Changchun are mainly viruses notably, respiratory syncytial virus is the dominant pathogen, followed by Mycoplasma pneumoniae.The bacterial pathogen is mainly Streptococcus pneumoniae.High fever, faster breathing, and cyanosis around the mouth are risk factors for severe pneumonia.Multi-pathogen mixed infection is prone to serious complications.
7.Analysis of respiratory pathogens for children respiratory tract infection by capillary electrophoresis﹣based multiplex PCR
Yuqi LIU ; Yanzhi HUANG ; Liwei SUN ; Hongbo JIANG ; Yuling TIAN ; Tingyu MENG ; Yan MA ; Yan HE ; Yanling ZHAO ; Yingwei MA
Chinese Pediatric Emergency Medicine 2019;26(10):764-770
Objective To investigate the effect of capillary electrophoresis﹣based multiplex PCR ( CEMP) in detecting pathogens for children respiratory tract infection,and to provide scientific basis for clin﹣ical diagnosis and treatment rapidly and accurately. Methods The cases were defined according to the na﹣tional monitoring program of febrile respiratory syndrome during the 12th Five﹣Year Plan,and the samples were collected from nasopharyngeal swabs,bronchoalveolar lavage fluid and sputum of children with respira﹣tory tract infection hospitalized in Changchun Children′s Hospital from January 2017 to February 2018. Multi﹣plex PCR amplification was performed by one﹣step method, then PCR products were separated by DNA length size with capillary electrophoresis and pathogens were analyzed by"Genemapper software" software. Detecting pathogens included Influenza A virus (InfA),Human Adenovirus (HADV),Boca virus ( Boca), Human Rhinovirus ( HRV), Novel InfA﹣09H1 ( InfA﹣09H1 ) and Seasonal Influenza virus H3N2 ( InfA﹣H3N2),Parainfluenza virus ( HPIV),Human metapneumonia virus ( HMPV), Influenza B virus ( InfB), Mycoplasma pneumoniae (Mp),Chlamydia pneumoniae ( CP),Human Coronavirus ( HCOV),Human Re﹣spiratory Syncytial virus (HRSV). Results The effective detection rate of the CEMP assay was 95. 71%. The positive detection rate of respiratory tract pathogens was 62. 84% and the mixed infection rate was 9. 61%. The mixed infection was mainly InfA and HRSV. The highest three positive rates were named InfA, HRSV and Mp. The positive rate of HRSV was significantly higher in the 0﹣3 age group than that in older group. Different pathogens were detected in different age groups,and the high﹣occurrence season of respiratory tract infection with virus was from December to March of the next year. InfA﹣09H1 was the main prevalent influenza virus in January,February and March 2017,InfA﹣H3N2 was the main prevalent influenza virus in November and December 2017,and the outbreak of InfB was happened in Changchun in late 2017 and early 2018. HRSV was detected only in the coldest season in Changchun from November to March of the next year. Different pathogens were detected in different respiratory infection. HRSV was the main pathogen detec﹣ted in pneumonia; InfA﹣03H2 and HPIV were the main pathogens detected in acute bronchitis; HRV and InfA were the main pathogens detected in upper respiratory tract infection. Conclusion CEMP is an effi﹣cient,rapid and accurate method for the detection of pathogens in patients with respiratory tract infections,and it will have a broad application prospect to develop reagents suitable for clinical diagnosis.
8. Analysis of respiratory pathogens for children respiratory tract infection by capillary electrophoresis-based multiplex PCR
Yuqi LIU ; Yanzhi HUANG ; Liwei SUN ; Hongbo JIANG ; Yuling TIAN ; Tingyu MENG ; Yan MA ; Yan HE ; Yanling ZHAO ; Yingwei MA
Chinese Pediatric Emergency Medicine 2019;26(10):764-770
Objective:
To investigate the effect of capillary electrophoresis-based multiplex PCR (CEMP) in detecting pathogens for children respiratory tract infection, and to provide scientific basis for clinical diagnosis and treatment rapidly and accurately.
Methods:
The cases were defined according to the national monitoring program of febrile respiratory syndrome during the 12th Five-Year Plan, and the samples were collected from nasopharyngeal swabs, bronchoalveolar lavage fluid and sputum of children with respiratory tract infection hospitalized in Changchun Children′s Hospital from January 2017 to February 2018.Multiplex PCR amplification was performed by one-step method, then PCR products were separated by DNA length size with capillary electrophoresis and pathogens were analyzed by "Genemapper software" software.Detecting pathogens included Influenza A virus (InfA), Human Adenovirus (HADV), Boca virus (Boca), Human Rhinovirus (HRV), Novel InfA-09H1 (InfA-09H1) and Seasonal Influenza virus H3N2 (InfA-H3N2), Parainfluenza virus (HPIV), Human metapneumonia virus (HMPV), Influenza B virus (InfB), Mycoplasma pneumoniae (Mp), Chlamydia pneumoniae (CP), Human Coronavirus (HCOV), Human Respiratory Syncytial virus (HRSV).
Results:
The effective detection rate of the CEMP assay was 95.71%.The positive detection rate of respiratory tract pathogens was 62.84% and the mixed infection rate was 9.61%.The mixed infection was mainly InfA and HRSV.The highest three positive rates were named InfA, HRSV and Mp.The positive rate of HRSV was significantly higher in the 0-3 age group than that in older group.Different pathogens were detected in different age groups, and the high-occurrence season of respiratory tract infection with virus was from December to March of the next year.InfA-09H1 was the main prevalent influenza virus in January, February and March 2017, InfA-H3N2 was the main prevalent influenza virus in November and December 2017, and the outbreak of InfB was happened in Changchun in late 2017 and early 2018.HRSV was detected only in the coldest season in Changchun from November to March of the next year.Different pathogens were detected in different respiratory infection.HRSV was the main pathogen detected in pneumonia; InfA-03H2 and HPIV were the main pathogens detected in acute bronchitis; HRV and InfA were the main pathogens detected in upper respiratory tract infection.
Conclusion
CEMP is an efficient, rapid and accurate method for the detection of pathogens in patients with respiratory tract infections, and it will have a broad application prospect to develop reagents suitable for clinical diagnosis.
9.Detection and homology analysis of virulence genes in pandrug-resistant Pseudo-monas aeruginosa
Huiling CHEN ; Hui SHENG ; Huifen YE ; Yingwei YANG ; Yiyan YANG ; Xiaoyuan HUANG ; Yinmei YANG
Chinese Journal of Infection and Chemotherapy 2013;(6):469-472
Objective To study the prevalence and sequence homology of virulence genes exoU and exoS in 53 strains of pan-drug-resistant Pseudomonas aeruginosa .Methods The virulence genes exoU and exoS were detected by PCR.Sequence homo-logy was analyzed by BOX-PCR.Results Of the 53 clinical isolates of pandrug-resistant Pseudomonas aeruginosa ,the exoS+/exoU- genotype was identified in 40 strains,exoU+/exoS - genotype in 10 strains,exoS +/exoU+ genotype in 1 strain, and exoS-/exoU- genotype in 2 strains.BOX-PCR results showed that 41 exoS+ isolates belonged to 24 genotypes,and 11 exoU+ strains could be grouped into 7 genotypes.Conclusions The prevalence of virulence genes is high in clinical isolates of pandrug-resistant Pseudomonas aeruginosa .BOX-PCR fingerprint analysis combined with sequence homology analysis is help-ful for effective monitoring and control of hospital pandrug-resistant pseudomonas aeruginosa infection.
10.Effect of Losartan and Amlodipine on serum and urine transforming growth factor-beta 1 in kidney transplantation recipients
Tongqing CHEN ; Yingwei HUANG ; Yaozhong KONG ; Minwa LIN ; Bihong WU ; Guiying LIAN
Chinese Journal of Tissue Engineering Research 2009;13(53):10541-10544
OBJECTIVE: To investigate the effect of Losartan and Amlodipine on serum and urine transforming growth factor -β_1 in patients undergoing kidney transplantation. METHODS: A total of 40 patients with mild or moderate hypertension (systolic pressure 140-170 mm Hg, and diastolic pressure 85-100 mm Hg, 1 mm Hg=0.133 kPa) following primary kidney transplantation were selected from First People's Hospital of Foshan, including 23 males and 17 females aged (38.6±19.2) years. They were randomly divided into two groups (n=20): Losartan group (oral administration 50 mg per day) and Amlodipine group (oral administration 5 mg per day). The blood pressure of patients should be controlled below 130/80 mm Hg. The blood pressure, renal function, 24 h-proteinuria, serum and urine transforming growth factor-β_1 6 months after medication were observed. RESULTS: A total of 40 patients were included in the final analysis. The systolic pressure and diastolic pressure of patients were decreased after administration (P < 0.05) and decreased to normal levels 6 months after administration (P < 0.01). During treatment, there were significant differences in blood pressure decrease and mean arterial pressure between two groups (P > 0.05). No difference was found in total efficacy between two groups (P > 0.05). In addition, blood urea nitrogen, creatinine, and blood uric acid did not significantly alter after treatment in two groups (P > 0.05). After 6 months of treatment, 24 h-proteinuria, serum and urine transforming growth factor -β_1 in Losartan group were significantly decreased compared with before treatment (P < 0.05), while no obvious changes were found in Amlodipine group (P > 0.05). The 24 h-proteinuria, serum and urine transforming growth factor-β_1 in Iosartan group were significantly less than Amlodipine group (P < 0.05).CONCLUSION: Both Losartan and Amlodipine effectively controlled hypertension of patients following kidney transplantation, but Losartan significantly decreased 24 h-proteinuria, serum and urine transforming growth factor-β_1 compared with Amlodipine.

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