1.Development and application of a triplex TaqMan fluorescent quantitative PCR assay for simultaneous detection of Senecavirus A,foot-and-mouth disease virus and porcine teschovirus
Shiqi GAN ; Qianhe WEI ; Yuchen NI ; Jianbo NI ; Xiuling ZHAO ; Wanyu DONG ; Yings-han ZHOU ; Xiaodu WANG
Chinese Journal of Veterinary Science 2025;45(1):22-29
Primers and probes were designed based on the conserved regions of Senecavirus A(SVA),foot-and-mouth disease virus(FMDV),and porcine teschovirus(PTV)and used to devel-op a TaqMan fluorescent quantitative PCR method for detecting the above-mentioned three viru-ses.The triplex fluorescent quantitative PCR system was developed using recombinant positive plasmids containing conserved sequences of the three viruses as templates.After optimizing the conditions,the specificity,sensitivity,repeatability,standard curve,and mixed infection model were evaluated,and the constructed triplex fluorescent quantitative PCR was used for clinical detection.The results showed that this method could specifically detect SVA,FMDV and PTV without cross-reactivity with other pathogens with the minimal detection concentrations for SVA,FMDV,and PTV as low as 1X101 copies/μL,respectively.The coefficients of variation within and between groups were less than 5%.Furthermore,none of the three viruses were detected in 126 samples.The above results indicate that this method is highly specific,sensitive,and stable,making it suit-able for clinical detection.
2.Development and application of a triplex TaqMan fluorescent quantitative PCR assay for simultaneous detection of Senecavirus A,foot-and-mouth disease virus and porcine teschovirus
Shiqi GAN ; Qianhe WEI ; Yuchen NI ; Jianbo NI ; Xiuling ZHAO ; Wanyu DONG ; Yings-han ZHOU ; Xiaodu WANG
Chinese Journal of Veterinary Science 2025;45(1):22-29
Primers and probes were designed based on the conserved regions of Senecavirus A(SVA),foot-and-mouth disease virus(FMDV),and porcine teschovirus(PTV)and used to devel-op a TaqMan fluorescent quantitative PCR method for detecting the above-mentioned three viru-ses.The triplex fluorescent quantitative PCR system was developed using recombinant positive plasmids containing conserved sequences of the three viruses as templates.After optimizing the conditions,the specificity,sensitivity,repeatability,standard curve,and mixed infection model were evaluated,and the constructed triplex fluorescent quantitative PCR was used for clinical detection.The results showed that this method could specifically detect SVA,FMDV and PTV without cross-reactivity with other pathogens with the minimal detection concentrations for SVA,FMDV,and PTV as low as 1X101 copies/μL,respectively.The coefficients of variation within and between groups were less than 5%.Furthermore,none of the three viruses were detected in 126 samples.The above results indicate that this method is highly specific,sensitive,and stable,making it suit-able for clinical detection.
4.Analysis of clinical characteristics of thyroid carcinoma in 57 flying personnel
Zhiyuan MAO ; Yu WANG ; Juqin LIU ; Haiyan YU ; Xiuling XIA ; Lulu SUN ; Hui ZHAO ; Zaiwen FAN
Chinese Journal of Aerospace Medicine 2023;34(2):85-89
Objective:To improve the diagnosis and treatment of thyroid carcinoma in flying personnel by analyzing the clinical characteristics of thyroid carcinoma.Methods:The clinical and pathological data of flying personnel with thyroid carcinoma diagnosed and treated by Air Force Medical Center of PLA from January of 2010 to December of 2022 were retrospectively analyzed. Clinical characteristics such as age, flying hours, aircraft types, thyroid function at diagnosis, and pathological characteristics such as tumor site, tumor size, lymph node metastasis, calcification and sand and Ki-67 positive ratio were collected, and the clinical and pathological characteristics were statistically analyzed.Results:Thyroid nodule was found as the first diagnosis of thyroid carcinoma in 57 flying personnel. The age of onset was 22-58 years old, the median age was 37 years old, the flying hours was 4-18 000 h, and the median flying hours was 2 000 h. Thyroid carcinoma was detected in both thyroid glands, and the histological type was papillary thyroid carcinoma. Thyroid function was normal in most cases when thyroid carcinoma was detected. Lymph node metastasis of thyroid carcinoma was low positively correlated with tumor size ( r=0.304, P=0.021), and was low positively correlated with the positive proportion of Ki-67 ( r=0.360, P=0.006). Other clinical and pathological characteristics did not show clear correlation. With the extension of flying hours, the incidence of lymph node metastasis of thyroid carcinoma increased, and the difference was statistically significant ( χ2=6.32, P=0.012). There was no statistical difference among the clinical characteristics of flying personnel with different age and aircraft types ( P>0.05). Conclusions:The thyroid carcinoma is usually diagnosed without clinical symptoms and the thyroid function is basically normal. The thyroid ultrasound examination should be emphasized during physical examination, and further examination should be conducted for the flying personnel with thyroid nodules.
5.Analysis of clinical characteristics of thyroid carcinoma in 57 flying personnel
Zhiyuan MAO ; Yu WANG ; Juqin LIU ; Haiyan YU ; Xiuling XIA ; Lulu SUN ; Hui ZHAO ; Zaiwen FAN
Chinese Journal of Aerospace Medicine 2023;34(2):85-89
Objective:To improve the diagnosis and treatment of thyroid carcinoma in flying personnel by analyzing the clinical characteristics of thyroid carcinoma.Methods:The clinical and pathological data of flying personnel with thyroid carcinoma diagnosed and treated by Air Force Medical Center of PLA from January of 2010 to December of 2022 were retrospectively analyzed. Clinical characteristics such as age, flying hours, aircraft types, thyroid function at diagnosis, and pathological characteristics such as tumor site, tumor size, lymph node metastasis, calcification and sand and Ki-67 positive ratio were collected, and the clinical and pathological characteristics were statistically analyzed.Results:Thyroid nodule was found as the first diagnosis of thyroid carcinoma in 57 flying personnel. The age of onset was 22-58 years old, the median age was 37 years old, the flying hours was 4-18 000 h, and the median flying hours was 2 000 h. Thyroid carcinoma was detected in both thyroid glands, and the histological type was papillary thyroid carcinoma. Thyroid function was normal in most cases when thyroid carcinoma was detected. Lymph node metastasis of thyroid carcinoma was low positively correlated with tumor size ( r=0.304, P=0.021), and was low positively correlated with the positive proportion of Ki-67 ( r=0.360, P=0.006). Other clinical and pathological characteristics did not show clear correlation. With the extension of flying hours, the incidence of lymph node metastasis of thyroid carcinoma increased, and the difference was statistically significant ( χ2=6.32, P=0.012). There was no statistical difference among the clinical characteristics of flying personnel with different age and aircraft types ( P>0.05). Conclusions:The thyroid carcinoma is usually diagnosed without clinical symptoms and the thyroid function is basically normal. The thyroid ultrasound examination should be emphasized during physical examination, and further examination should be conducted for the flying personnel with thyroid nodules.
6.The clinical characteristic and risk factors of subcutaneous tophus among hospitalized gout patients
Linling SONG ; Xiuling NIE ; Junwei WANG ; Maoyue LI ; Wei ZHAO
Chinese Journal of Postgraduates of Medicine 2022;45(7):581-587
Objective:To explore the clinical characteristic and risk factors of subcutaneous tophus among hospitalized gout patients.Methods:The clinical data of 646 inpatients with gout from April 2014 to December 2019 in Tianjin Medical University Chu Hsien-I Memorial Hospital were retrospectively analyzed. The patients were divided into tophus group (172 cases) and non-tophus group (474 cases) according to the presence of subcutaneous tophus. The body height, body mass, waist circumference, hip circumference and blood pressure were measured. The total cholesterol (TC), triglyceride (TG), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), urea nitrogen, creatinine, uric acid and C-reactive protein (CRP) were detected; urine was collected for 24 h to detect uric acid. The body mass index (BMI), estimated glomerular filtration rate (eGFR) and uric acid excretion fraction (FUA) were counted. Binary Logistic regression analysis was used to analyze the independent risk factors of subcutaneous tophus among hospitalized gout patients.Results:The proportion of male, course of disease, number of joint involved in attack, rate of smoking history, rate of drinking history, smoking amount, alcohol consumption volume, length of time spent drinking, length of time spent smoking, LDL-C, urea nitrogen, serum creatinine and serum uric acid in tophus group were significantly higher than those in non-tophus group: 98.84% (170/172) vs. 94.09% (446/474), 10 (5, 16) years vs. 5 (2, 9) years, 6 (4, 8) joints vs. 3 (2, 5) joints, 66.86% (115/172) vs. 58.44% (277/474), 65.70% (113/172) vs. 57.38% (272/474), 11 (0, 20) cigarettes /d vs. 10 (0, 20) cigarettes/d, 100 (0, 250) ml/d vs. 50 (0, 162) ml/d, 10 (0, 26) years vs. 0 (0, 20) years, 20 (0, 30) years vs. 10 (0, 20) years, (3.44 ± 0.98) mmol/L vs. (3.25 ± 0.97) mmol/L, 5.81 (4.61, 7.46) mmol/L vs. 5.38 (4.39, 6.66) mmol/L, 89.4 (74.3, 107.5) μmol/L vs. 85.1 (72.5, 98.9) μmol/L and 514.4 (452.9, 595.2) μmol/L vs. 499.0 (404.8, 572.4), the onset age, red blood cell, hemoglobin and eGFR were significantly lower than those in non-tophus group: (37.78 ± 10.56) years old vs. (40.17 ± 13.06) years old, (4.72 ± 0.74) × 10 9/L vs. (4.88 ± 0.56) × 10 9/L, (141.15 ± 17.19) g/L vs. (146.00 ± 13.06) g/L and 87.7 (65.9, 108.4) ml/min vs. 93.3 (75.9, 113.1) ml/min, and there were statistical differences ( P<0.05 or <0.01); there were no statistical differences in family history of diabetes, family history of hypertension, family history of gout, BMI, waist-hip ratio, blood pressure, FUA, white blood cell, TC, TG, HDL-C and CRP between 2 groups ( P>0.05). The patients were grouped according to the course of disease, the incidences of tophus in patients with course of disease ≤5.0 years, 5.1 to 10.0 years and >10.0 years were 15.34% (50/326), 27.67% (44/159) and 48.45% (78/161), and there was statistical difference ( χ2 = 60.59, P<0.01); the patients were grouped according to the quartiles of serum uric acid, the incidences of tophus in patients with serum uric acid <424.05 μmol/L, 424.05 to 505.24 μmol/L, 505.25 to 576.17 μmol/L and ≥576.18 μmol/L were 14.91% (24/161), 32.72% (53/162), 29.01% (47/162) and 29.81% (48/161), and there was statistical difference ( χ2 = 15.70, P<0.01). Binary Logistic regression analysis result showed that the course of gout, smoking amount, number of joint involved in attack, serum uric acid and LDL-C were the independent risk factors of subcutaneous tophus among hospitalized gout patients ( OR = 1.069, 1.020, 1.317, 1.002 and 1.262; 95% CI 1.032 to 1.097, 1.006 to 1.032, 1.223 to 1.417, 1.000 to 1.003 and 1.033 to 1.541; P<0.01 or <0.05). Conclusions:The duration of disease, number of involved joint, serum uric acid level, unhealthy lifestyles and lipid metabolism disorders are the independent risk factors of subcutaneous tophus among hospitalized gout patients.
7.Short-term effects of acute fructose intake on blood antioxidant capacity and liver enzymes in healthy young adults
Xiuling NIE ; Jing ZHANG ; Linling SONG ; Wei ZHAO ; Xiaocui CHENG ; Pei SUN
Chinese Journal of Postgraduates of Medicine 2022;45(7):587-593
Objective:To investigate the short-term effects of acute fructose intake on serum antioxidant capacity and liver enzymes in healthy young adults.Methods:From January to June 2019, 64 healthy young subjects were recruited, and divided into 75 g glucose group, 25 g fructose group, 50 g fructose group and 75 g fructose group by random digits table method with 16 cases each. The subjects took corresponding amounts of glucose or fructose according to grouping. The levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), glutathione peroxidase (GPX), superoxide dismutase (SOD), C-Jun amino terminal kinase (JNK), malondialdehyde (MDA) and 8-OH deoxyguanine (8-OHdG) before taking sugar and 30, 60, 120, 180 min after taking sugar, and the changes of ALT, AST and LDH at 30, 60, 120 and 180 min after taking sugar compared with that before taking sugar.Results:One case in 50 g fructose group, 2 cases in 75 g fructose group and 1 case in 75 g glucose group dropped out due to adverse reaction; finally, 15 cases in 75 g glucose group, 16 cases in 25 g fructose group, 15 cases in 50 g fructose group and 14 cases in 75 g fructose group completed the study. The increase of ALT and AST after taking sugar in 25 g fructose group, 50 g fructose group and 75 g fructose group was significantly higher than that in 75 g glucose group, and there were statistical differences ( P<0.05); there was no statistical difference in the change of LDH after taking sugar among 4 groups ( P>0.05). One hundred and eighty min after taking sugar, the receiver operating characteristic (ROC) curve analysis result showed that there were no statistical differences in the areas under curve of ALT, AST and LDH among 4 groups ( P>0.05). There was no statistical difference in SOD before taking sugar among 4 groups ( P>0.05); the SOD 60 min after taking sugar in 50 g fructose group and 75 g fructose group, and SOD 180 min after taking sugar in 25 g fructose group, 50 g fructose group and 75 g fructose group were significantly lower than those in 75 g glucose group: (4.84 ± 1.88) and (4.38 ± 1.12) μg/L vs. (6.25 ± 1.65) μg/L, (4.46 ± 1.66), (5.22 ± 1.66) and (3.99 ± 0.96) μg/L vs. (6.55 ± 1.78) μg/L, and there were statistical differences ( P<0.05). There were no statistical differences in the changes of JNK, GPX, MDA and 8-OHdG before and after taking sugar among 4 groups ( P>0.05). The ROC curve 180 min after taking sugar analysis result showed that the area under curve of SOD in 75 g fructose group was significantly lower than that in 75 g glucose group (9.06 ± 1.88 vs. 12.74 ± 3.15), and there was statistical difference ( P<0.05); there were no statistical differences in the areas under curve of GPX, JNK, MDA and 8-OHdG among 4 groups ( P>0.05). Conclusions:Acute fructose intake can lead to the decrease of antioxidant capacity, and the increasing of oxidative damage and liver enzymes in healthy adults.
8.Analysis of the salt-stress responsive element of the promoter of peanut small GTP binding protein gene AhRabG3f.
Guoning DU ; Jie XIANG ; Shunyu LIN ; Xiangyuan KONG ; Xiuling WU ; Xuedong GUAN ; Hong ZHU ; Jingshan WANG ; Lixian QIAO ; Jiongming SUI ; Chunmei ZHAO
Chinese Journal of Biotechnology 2022;38(8):2989-2998
To study the molecular mechanism of salt stress response of peanut small GTP binding protein gene AhRabG3f, a 1 914 bp promoter fragment upstream of the start codon of AhRabG3f gene (3f-P) from peanut was cloned. Subsequently, five truncated fragments (3f-P1-3f-P5) with lengths of 1 729, 1 379, 666, 510 and 179 bp were obtained through deletion at the 5' end, respectively. Plant expression vectors where these six promoter fragments were fused with the gus gene were constructed and transformed into tobacco by Agrobacterium-mediated method, respectively. GUS expression in transgenic tobacco and activity analysis were conducted. The gus gene expression can be detected in the transgenic tobacco harboring each promoter segment, among which the driving activity of the full-length promoter 3f-P was the weakest, while the driving activity of the promoter segment 3f-P3 was the strongest. Upon exposure of the transgenic tobacco to salt stress, the GUS activity driven by 3f-P, 3f-P1, 3f-P2 and 3f-P3 was 3.3, 1.2, 1.9 and 1.2 times compared to that of the transgenic plants without salt treatment. This suggests that the AhRabG3f promoter was salt-inducible and there might be positive regulatory elements between 3f-P and 3f-P3 in response to salt stress. The results of GUS activity driven by promoter fragments after salt treatment showed that elements included MYB and GT1 between 1 930 bp and 1 745 bp. Moreover, a TC-rich repeat between 682 bp and 526 bp might be positive cis-elements responsible for salt stress, and an MYC element between 1 395 bp and 682 bp might be a negative cis-element responsible for salt stress. This study may facilitate using the induced promoter to regulate the salt resistance of peanut.
Arachis/genetics*
;
Fabaceae/genetics*
;
GTP-Binding Proteins/metabolism*
;
Gene Expression Regulation, Plant
;
Glucuronidase/metabolism*
;
Plant Proteins/metabolism*
;
Plants, Genetically Modified/genetics*
;
Salt Stress
;
Stress, Physiological/genetics*
;
Tobacco/genetics*
9.Protective effect of Acronychia pedunculata water extract on photoaging keratinocyte model and its mechanism
Xiuling LUO ; Yuan ZHOU ; Binbin ZHAO ; Manli HUANG ; Wenyu LI ; Runge FAN ; Sijian WEN ; Jichao LI ; Huayu WU ; Youkun LIN
Chinese Journal of Medical Aesthetics and Cosmetology 2022;28(6):517-521
Objective:To investigate the protective effect and mechanism of Acronychia pedunculata water extracts on UV-induced light damage of human keratinocytes.Methods:The experiment was conducted from December 2018 to April 2020 in the Guangxi Medical University Laboratory of Genetics. The photoaged keratinocyte model was used, the cells were co-cultured with different concentrations of Acronychia pedunculata water extracts. The cell proliferation rate was detected by CCK-8 method. The levels of reactive oxygen species (ROS), malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX) and total antioxidant capacity (T-AOC) of cells were detected by a test kit. The levels of IL-1β, IL-6 and tumor necrosis factor-alpha (TNF-α) were determined by ELISA.Results:The proliferation of HaCaT cells was promoted by 0.5 mg/L-2.0 mg/L of the extracts. Compared with control group, the proliferation rate of HaCaT cells in the experimental group was significantly increased ( P<0.05). Compared with control group, the contents of ROS was decreased ( F=214.67, P<0.05), MDA was decreased ( F=811.88, P<0.05), SOD was increased ( F=28.95, P<0.05), CAT was increased ( F=213.31, P<0.05), GPX was increased ( F=65.10, P<0.05), T-AOC was increased ( F=305.58, P<0.05), IL-1β was decreased ( F=15.46, P<0.05), IL-6 was decreased ( F=59.2, P<0.05), and TNF-α was decreased ( F=33.13, P<0.05). Conclusions:The extracts of 0.5-2.0 mg/L of Acronychia pedunculata have protective effects on the photoaging cell model, which may be related to the increase of SOD, CAT, GPX and other antioxidant enzymes and the level of T-AOC in photoaging HaCaT cells, and the decrease of ROS, MDA content and the expression of inflammatory cytokines.
10.Variation of the Vaginal Microbiome During and After Pregnancy in Chinese Women
Zhang XIAOAI ; Zhai QINGZHI ; Wang JINFENG ; Ma XIULING ; Xing BO ; Fan HANG ; Gao ZHIYING ; Zhao FANGQING ; Liu WEI
Genomics, Proteomics & Bioinformatics 2022;20(2):322-333
A comprehensive profiling of the vaginal microbial communities and their variability enables an accurate description of the microbiome in women.However,there is a lack of studies available on Chinese women.In the present study,the composition of the vaginal microbiota during pregnancy and the 6-week postpartum period of 454 Chinese women was characterized by sequenc-ing the V3-V4 region of the 16S ribosomal RNA(rRNA)gene.The vaginal microbiome showed variations during pregnancy and the postpartum period based on the abortion history,hypertensive disorders,delivery mode,and maternal age.Co-variation of 22 bacterial taxa,including the Lacto-bacillus genus and two of its species,may account for the common characteristics of the vaginal microbiome under scenarios of different medical histories and pregnancy outcomes.In contrast,dis-criminant bacterial species were significantly different between women who had preterm birth(PTB)with and without premature rupture of membranes(PROM),and the community state type(CST)Ⅳ-A without any predominant Lactobacillus species in the microbiota was more prevalent during pregnancy in the PROM-PTB cases,suggesting that specific bacterial species could be considered to distinguish between different types of PTB.By providing data on Chinese women,this study will enrich the knowledge of the human microbiome and contribute to a better understanding of the association between the vaginal microbiome and reproductive health.

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