1.Establishment of SYBR Green Ⅰ-based quantitative PCR targeting pseudorabies virus UL35 gene
Xinyu ZHANG ; Hongxia WU ; Yongfeng LI ; Yuan SUN ; Qiang FU ; HuaJi QIU
Chinese Journal of Veterinary Science 2024;44(11):2334-2340
Quantitative analysis of UL35 and gB genes at different time points after PRV infection showed that there were significant differences between the two at 2.5,5.0 and 20.0 h after infec-tion,and the expression of UL35 gene could be observed in the early stage of PRV infection.To de-termine whether the UL 35 gene can be used as a target for the diagnosis of PRV infection,a spe-cific primer pair was designed and synthesized according to the conserved sequence of the UL35 gene of different PRV strains,and used to amplify a fragment of 54 bp in length.After optimizing the reaction conditions and system,the standard curve for the established by SYBR Green Ⅰ real-time PCR detection method showed that it had good repeatability,specificity and a good linear rela-tionship to the template concentration.No amplifications for porcine reproductive and respiratory syndrome virus(PRRSV),classical swine fever virus(CSFV),and African swine fever virus(AS-FV)were detected.Compared with the existing similar methods,the established method showed no difference in sensitivity.The coefficient of variation of inter-and intra-group repeatability for the method was less than 2%.Detection of the samples in mice challenged with PRV revealed a certain amount of viral load in tissue.The results showed that UL35 gene can be used as a target for the diagnosis of PRV infection.
2.Role and Mechanism of UBE2C in the Proliferation of MCF-7 Cells in Breast Cancer
Journal of Medical Research 2024;53(7):73-78
Objective To study the effect of silencing ubiquitin-conjugating enzyme 2C(UBE2C)on the proliferative capacity of human breast cancer cells MCF-7 and to preliminarily investigate its mechanism of action.Methods Silencing of UBE2C gene in MCF-7 cells using small interfering RNA(si-RNA).CCK-8 assay and clone formation assay were used to detect cell proliferation a-bility.The expression of cell cycle-associated protein kinase inhibitor protein p21 as well as cell cycle-promoting proteins CyclinD1,cyclin-dependent kinase 4(CDK4),and cyclin-dependent kinase 6(CDK6)were detected by real-time quantitative polymerase chain reaction(RT-qPCR)and Western blot at the mRNA and protein levels,respectively.Results Compared with the cells in blank control group and negative control(si-NC)group,the expression of UBE2C was down-regulated in cells of si-UBE2C group.The re-sults of CCK-8 assay and clone formation assay showed that the proliferative ability of cells in si-UBE2C group was significantly de-creased.Compared with the cells in blank control group and si-NC group,the expression of cell cycle-related protein kinase inhibitor protein p21 in the cells of si-UBE2C group was up-regulated at both the mRNA and protein levels,while the expression of cell cycle-promoting proteins CyclinD1,CDK4 and CDK6 was down-regulated at both the mRNA and protein levels.Conclusion UBE2C can reg-ulate the proliferative capacity of MCF-7 cells and promote breast cancer progression by mediating the expression of CyclinD1/CDK4/CDK6 complex and p21.
3.Surgical strategy for treatment of complex aortic coarctation
Xiaohan XU ; Miti WU ; Faliang ZHAN ; Tao YE ; Yizhou GAO ; Weidong GU ; Qun GU ; Yongfeng SHAO
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2024;31(05):776-781
Objective To summarize the clinical experience of the treatment for complex aortic coarctation with extra anatomic bypass and anatomic correction techniques. Methods The clinical data of patients with complex aortic coarctation treated in the First Affiliated Hospital of Nanjing Medical University and Friendship Hospital of Ili Kazakh Autonomous Prefecture between April 2012 and November 2020 were retrospectively reviewed. Results A total of 12 patients were enrolled, including 5 males and 7 females aged 11-54 (34.3±16.2) years. Extra anatomic bypass grafting was performed in 8 patients and anatomic correction was performed in 4 patients. The operations were successful in all patients. There was no perioperative death. The average cardiopulmonary bypass time was 203.0±46.0 min (7 median incision patients), and the average intraoperative blood loss was 665.0±102.0 mL. The average postoperative ventilator support time was 32.3±7.5 h, and the average postoperative hospital stay time was 10.2±4.3 d. The mean drainage volume of median incision was 1 580.0±360.0 mL, and the mean drainage time was 9.3±2.7 d. The mean drainage volume of left thoracotomy was 890.0±235.0 mL, and the mean drainage time was 4.8±2.5 d. One patient had a transient hoarse after operation and recovered 6 months later. The follow-up period ranged from 2 to 10 years with an average time of 81.0±27.0 months. All patients had a recovery of hypertension, cardiac afterload after 2 years postoperatively. One patient who received an artificial blood vessel replacement in situ was examined stenosis recurrence at the third year after discharge. Others were asymptomatic during the follow-up period. There were no death or other complications. Conclusion The treatment strategy for complex aortic coarctation should be individualized according to the anatomical features and concomitant heart diseases. Extra anatomic bypass technique is a safe and feasible choice.
4.Combination of serum alpha-fetoprotein,PIVKA-Ⅱ and glypican-3 in diagnosis of hepatocellular carcinoma:a meta-analysis
Hongliang SONG ; Jianguo WANG ; Hui ZHANG ; Yongfeng WU ; Kai WANG ; Xiaobo WANG ; Xiao XU
Journal of Zhejiang University. Medical sciences 2024;53(1):131-139
Objective:To assess the value of serum alpha-fetoprotein(AFP),protein induced by vitamin K absence or antagonist-Ⅱ(PIVKA-Ⅱ)and glypican-3(GPC-3)in the diagnosis of hepatocellular carcinoma(HCC).Methods:Studies of AFP,PIVKA-Ⅱ,GPC-3 or in combination for the diagnosis of HCC since 2002 were searched in PubMed,Web of Science and Embase databases.The literature was screened according to the inclusion and exclusion criteria,the quality of the included articles was evaluated by QUADAS checklist,and relevant data were extracted by Meta DiSc,Review Manager 5.4 and Stata 15.1.The diagnostic values of AFP,PIVKA-Ⅱ and GPC-3 alone or in combination for HCC were assessed with receiver operating characteristic(ROC)curve.Results:A total of 32 articles were included in the study.Meta-analysis showed that when a single marker was used to diagnose HCC,the area under the ROC curve(AUC)of PIVKA-Ⅱ was the highest(0.88,95%CI:0.85-0.91),followed by GPC-3 and AFP.The AUC of combination of serum markers was higher than that of a single marker,and the AUC of PIVKA-Ⅱ combined with GPC-3 was the highest(0.90,95%CI:0.87-0.92).When a single marker was used for diagnosis,the sensitivity of PIVKA-Ⅱ and GPC-3 were relatively high(0.75 and 0.76),while the specificity of PIVKA-Ⅱ(0.88)and AFP(0.87)were higher than that of GPC-3(0.81).The sensitivity of the combination of serum markers was higher than that of a single marker,while the specificity was not significantly improved.When a single marker is used to diagnose HCC,the diagnostic odds ratio(DOR)of PIVKA-Ⅱ was the highest(22,95%CI:13-36),followed by GPC-3 and AFP.The DOR of the combination of two markers in the diagnosis of HCC was higher than that of a single marker,and the DOR of AFP combined with GPC-3 was the highest(25,95%CI:9-67).The DOR of the combination of the three markers was significantly reduced to 10(95%CI:7-45).Conclusions:When a single marker is used,PIVKA-Ⅱhas a higher diagnostic value for HCC.The combination of two markers can significantly improve the diagnostic sensitivity,and AFP combined with PIVKA-Ⅱ is recommended for the diagnosis of HCC.The combination of all three markers failed to further improve the diagnostic value.
5.Changes of median nerve F-wave parameters after manual digitorum sensory stimulation of hemiplegic fingers in stroke patients
Yu WU ; Yanjie HU ; Ziyan HE ; Yaxing TIAN ; Yongfeng HONG
Chinese Journal of Neuromedicine 2023;22(3):274-278
Objective:To evaluate the influence of manual digitorum sensory stimulation (MDSS) of hemiplegic fingers in median nerve F-wave in stroke patients.Methods:Thirty patients with hemiplegia after stroke, admitted to Department of Rehabilitation Medicine, Second Affiliated Hospital of Anhui Medical University from June 2019 to October 2022 were selected; all patients had thumb flexor modified Ashworth scale (MAS) grading≥1. Hemiplegic MDSS was given; bilateral median nerve F-wave before MDSS and median nerve F-wave at the hemiplegic side immediately after MDSS were recorded.Results:Compared with that at the healthy side before MDSS, amplitude of median nerve F-wave at the hemiplegic side was significantly increased ( P<0.05). Compared with that before MDSS, amplitude of F-wave at the hemiplegic side after MDSS in patients with thumb flexor MAS grading≤2 were significantly decreased ( P<0.05), while that in patients with thumb flexor MAS grading≥3 was significantly increased ( P<0.05). Conclusion:Amplitudes of median nerve F-wave at the hemiplegic side in patients with different thumb flexor MAS grading show that MDSS can not only inhibit the excitability of tonic motor unit related to muscle spasm, but also enhance the excitability of kinetic-motor unit mainly related to explosive power in severe muscle spasm.
6.Influencing factors of lower urinary tract symptoms in patients with prostate hyperplasia after transurethral holmium laser enucleation
Ting XU ; Junbiao HU ; Yongfeng ZHU ; Huiling WU
Chinese Journal of General Practitioners 2023;22(7):732-735
A total of 360 patients with prostate hyperplasia underwent transurethral holmium laser enucleation of the prostate in Department of Urology of Jinhua People′s Hospital from July 2019 to December 2022. Among 360 patients, lower urinary tract symptoms occurred one month after operation in 40 cases with an incidence rate of 11.11%. The age, body mass index (BMI), course of disease, nature of disease, preoperative prostate volume, postoperative prostate volume, preoperative IPSS score, postoperative IPSS score, preoperative maximum urine flow rate, postoperative maximum urine flow rate, preoperative residual urine volume, postoperative residual urine volume, postoperative urinary tract infection, operative time, postoperative catheter retention time were compared between patients with lower urinary tract symptoms (study group) and those without (control group). There were significant differences in the course of disease, preoperative prostate volume, preoperative IPSS score, preoperative maximum urine flow rate, preoperative residual urine volume, postoperative urinary tract infection and postoperative catheter retention time between two groups ( t=28.01, 6.35, 8.79, 17.92, 34.84, 11.45;all P<0.05). Multivariate logistic analysis showed that the course of disease, preoperative prostate volume, preoperative IPSS score, postoperative urinary tract infection, and postoperative catheter retention time were independent risk factors for postoperative lower urinary tract symptoms ( OR=6.964, 3.442, 1.944, 4.836, 4.225, 4.894; all P<0.05); while the preoperative maximum urinary flow rate was the protective factor( OR=0.043, P<0.05). The incidence of postoperative lower urinary tract symptoms in patients undergoing transurethral holmium laser enucleation of prostate is high. Effective protective measures should be taken based on the risk factors to reduce the incidence of postoperative lower urinary tract symptoms and to promote the early recovery of patients.
7.Diagnostic value of a combined serology-based model for minimal hepatic encephalopathy in patients with compensated cirrhosis
Shanghao LIU ; Hongmei ZU ; Yan HUANG ; Xiaoqing GUO ; Huiling XIANG ; Tong DANG ; Xiaoyan LI ; Zhaolan YAN ; Yajing LI ; Fei LIU ; Jia SUN ; Ruixin SONG ; Junqing YAN ; Qing YE ; Jing WANG ; Xianmei MENG ; Haiying WANG ; Zhenyu JIANG ; Lei HUANG ; Fanping MENG ; Guo ZHANG ; Wenjuan WANG ; Shaoqi YANG ; Shengjuan HU ; Jigang RUAN ; Chuang LEI ; Qinghai WANG ; Hongling TIAN ; Qi ZHENG ; Yiling LI ; Ningning WANG ; Huipeng CUI ; Yanmeng WANG ; Zhangshu QU ; Min YUAN ; Yijun LIU ; Ying CHEN ; Yuxiang XIA ; Yayuan LIU ; Ying LIU ; Suxuan QU ; Hong TAO ; Ruichun SHI ; Xiaoting YANG ; Dan JIN ; Dan SU ; Yongfeng YANG ; Wei YE ; Na LIU ; Rongyu TANG ; Quan ZHANG ; Qin LIU ; Gaoliang ZOU ; Ziyue LI ; Caiyan ZHAO ; Qian ZHAO ; Qingge ZHANG ; Huafang GAO ; Tao MENG ; Jie LI ; Weihua WU ; Jian WANG ; Chuanlong YANG ; Hui LYU ; Chuan LIU ; Fusheng WANG ; Junliang FU ; Xiaolong QI
Chinese Journal of Laboratory Medicine 2023;46(1):52-61
Objective:To investigate the diagnostic accuracy of serological indicators and evaluate the diagnostic value of a new established combined serological model on identifying the minimal hepatic encephalopathy (MHE) in patients with compensated cirrhosis.Methods:This prospective multicenter study enrolled 263 compensated cirrhotic patients from 23 hospitals in 15 provinces, autonomous regions and municipalities of China between October 2021 and August 2022. Clinical data and laboratory test results were collected, and the model for end-stage liver disease (MELD) score was calculated. Ammonia level was corrected to the upper limit of normal (AMM-ULN) by the baseline blood ammonia measurements/upper limit of the normal reference value. MHE was diagnosed by combined abnormal number connection test-A and abnormal digit symbol test as suggested by Guidelines on the management of hepatic encephalopathy in cirrhosis. The patients were randomly divided (7∶3) into training set ( n=185) and validation set ( n=78) based on caret package of R language. Logistic regression was used to establish a combined model of MHE diagnosis. The diagnostic performance was evaluated by the area under the curve (AUC) of receiver operating characteristic curve, Hosmer-Lemeshow test and calibration curve. The internal verification was carried out by the Bootstrap method ( n=200). AUC comparisons were achieved using the Delong test. Results:In the training set, prevalence of MHE was 37.8% (70/185). There were statistically significant differences in AMM-ULN, albumin, platelet, alkaline phosphatase, international normalized ratio, MELD score and education between non-MHE group and MHE group (all P<0.05). Multivariate Logistic regression analysis showed that AMM-ULN [odds ratio ( OR)=1.78, 95% confidence interval ( CI) 1.05-3.14, P=0.038] and MELD score ( OR=1.11, 95% CI 1.04-1.20, P=0.002) were independent risk factors for MHE, and the AUC for predicting MHE were 0.663, 0.625, respectively. Compared with the use of blood AMM-ULN and MELD score alone, the AUC of the combined model of AMM-ULN, MELD score and education exhibited better predictive performance in determining the presence of MHE was 0.755, the specificity and sensitivity was 85.2% and 55.7%, respectively. Hosmer-Lemeshow test and calibration curve showed that the model had good calibration ( P=0.733). The AUC for internal validation of the combined model for diagnosing MHE was 0.752. In the validation set, the AUC of the combined model for diagnosing MHE was 0.794, and Hosmer-Lemeshow test showed good calibration ( P=0.841). Conclusion:Use of the combined model including AMM-ULN, MELD score and education could improve the predictive efficiency of MHE among patients with compensated cirrhosis.
8.Microsatellite instability and its relationship with clinicopathological characteristics of patients with endometrial carcinoma
Zhuailin DUAN ; Yan WU ; Ruixia ZHAO ; Yongfeng GUO ; Yao ZHANG ; Jing SUN ; Haixia JIA
Cancer Research and Clinic 2023;35(7):521-525
Objective:To explore the status of microsatellite instability (MSI) and its relationship with clinicopathological characteristics of patients with endometrial carcinoma.Methods:The clinical data of 365 patients with endometrial carcinoma who received surgery in Shanxi Province Cancer Hospital between January 2020 and December 2021 were retrospectively analyzed. Immunohistochemistry was used to detect the expressions of 4 DNA mismatch repair (MMR) proteins (MLH1, MSH2, MHS6, and PMS2), estrogen receptor (ER), progesterone receptor (PR), and p53 mutant protein in postoperative cancer tissue samples from 365 patients with endometrial carcinoma. All patients were divided into MSI group (1 or more non-expression of MMR protein) and microsatellite stability (MSS) group (4 proteins were all expressed), and the clinicopathological characteristics of patients in both groups were compared. φ efficient was used to analyze the correlation of MSI with ER, PR, p53 mutant protein expressions. Results:There were 72 cases (19.7%) in MSI group and 293 cases (80.3%) in MSS group; and the age of all patients was (53±19) years (21-83 years). There were statistically significant differences in the proportion of MSI patients in endometrial carcinoma patients with different age [>50 years vs. ≤50 years: 22.1% (61/276) vs. 12.4% (11/89)], tumor diameter [≤2 cm vs. > 2 cm: 25.9% (30/116) vs. 16.8% (42/249)], International Federation of Gynecology and Obstetrics (FIGO) staging [stage Ⅲ-Ⅳ vs. stage Ⅰ-Ⅱ: 31.1% (14/45) vs. 18.1% (58/320)], histological type [type Ⅰ vs. type Ⅱ: 21.7% (71/327) vs. 2.6% (1/38)] (all P < 0.05). There were no statistically significant differences in the proportion of MSI patients with different depth of invasion, degree of differentiation, lymph node metastasis, vascular involvement, and lesion location (all P > 0.05). Among 327 cases of type Ⅰendometrial carcinoma, 1 case was mucinous adenocarcinoma (MSS status), and the other 326 cases were endometrioid adenocarcinoma. Of the 72 patients with MSI, 71 cases were endometrioid carcinoma and the other was 1 of 3 mixed carcinomas in type Ⅱ endometrial carcinoma. There was a negative correlation between MSI and mutant p53 ( φ coefficient was -0.11, P = 0.031), and φ coefficient of the correlation of MSI with ER and PR was -0.03 and -0.06, while there were no statistically significant differences ( P value was 0.578 and 0.255, respectively). Conclusions:Endometrioid adenocarcinoma is the main type of endometrial cancer patients with MSI. MSI in endometrial cancer is correlated with age, FIGO staging, tumor diameter and histological type of patients, while negatively correlated with mutant p53.
9.Features in digitorum muscular tension and shear-wave elastography in hemiplegic patients with stroke before and after manual digitorum sensory stimulation
Ziyan HE ; Xiaoxiao TANG ; Yaxing TIAN ; Fan JIANG ; Xiuli KAN ; Xianshan SHEN ; Jing MAO ; Jun XU ; Xue LIU ; Jianxian WU ; Tingting WU ; Yongfeng HONG
Chinese Journal of Rehabilitation Theory and Practice 2022;28(2):204-211
Objective To demonstrate the changes in flexor digitorum and extensor digitorum tension in the affected hands with shear-wave elastography (SWE) before and after manual digitorum sensory stimulation (MDSS) in hemiplegic patients with stroke. Methods A total of 51 hemiplegic post-stroke inpatients in the Department of Rehabilitation Medicine in Second Hospital of Anhui Medical University from April to June, 2020, underwent MDSS completed by a researcher who used a bare thumb and index finger to squeeze each nail bed as well as the sides of each fingertip in the affected hand. The stimulation intensity was the minimum that could cause finger extension without obvious pain, and the interval between two stimulations was five to ten seconds. Muscular tension of the flexor digitorum superficialis, flexor digitorum profundus, flexor pollicis longus and extensor digitorum were assessed with modified Ashworth Scale (MAS) and shear-wave velocity (SWV) of SWE on the affected side before and immediately after MDSS. MAS score was -1 as low muscular tension. Results The MAS scores of all the muscles significantly reduced after MDSS (|Z| > 2.843, P < 0.001); while the changes of SWV were not significantly in all the muscles with initially MAS score of 0 or -1 (t < 1.052, P > 0.05), and it reduced in those muscles with initial MAS scores of one to three (t > 2.672, P < 0.05). The SWV were positively correlated with the MAS scores both before and after MDSS (r > 0.334, P < 0.05). Conclusion MDSS can effectively, immediately, and safely relieves muscle spasms of the flexor digitorum and facilitate active finger extension in the affected hand for hemiplegic patients with stroke. SWE is useful for quantitatively and objectively evaluating muscular tension in the affected hand for hemiplegic patients with stroke.
10.Effects of manual digitorum sensory stimulation on spasticity of fingers for stroke patients: a surface electromyography study
Yaxing TIAN ; Yongfeng HONG ; Xiuli KAN ; Xianshan SHEN ; Jing MAO ; Yan JIANG ; Ziyan HE ; Yu WU ; Wei HU ; Xiaoning SUN ; Shunyin HU
Chinese Journal of Rehabilitation Theory and Practice 2022;28(5):515-519
ObjectiveTo investigate the changes of surface electromyography (sEMG) of the flexors and extensors of the affected fingers after manual digitorum sensory stimulation (MDSS) in the hemiplegic patients after stroke. MethodsFrom April to August, 2020, 50 stroke patients in Department of Rehabilitation Medicine, the Second Hospital of Anhui Medical University accepted MDSS on the nail beds and the third knuckles of affected fingers, until the fingers extended actively. The tension of affected flexor pollicis brevis, flexor digitorum superficialis and extensor digitorum were assessed with modified Ashworth Scale (MAS) before and immediately after stimulation; while the root mean square (RMS) of sEMG of bilateral flexor pollicis brevis, flexor digitorum superficialis and extensor digitorum were recorded. ResultsThe MAS score of all the muscles decreased after stimulation (|Z| > 2.699, P < 0.01), while the RMS of affected extensor digitorum increased (t = -2.069, P < 0.05). Compared with the unaffected ones, RMS of affected flexor pollicis brevis and extensor digitorum decreased before and after stimulation (t > 2.450, P < 0.05). ConclusionMDSS may immediately relieve the spasm of flexors of hemiplegic fingers after stroke, which associates with the promoting muscle strength of the extensors.

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