1.Establishment and assessment of a visual detection method for nucleic acid of Afri-can swine fever virus
Xingqi LIU ; Yujie BAI ; Mengyao ZHANG ; Jingbo HUANG ; Guangliang LIU ; Yuanyuan LI ; Shuyi TAN ; Haili ZHANG ; Yan ZHANG ; Zongxi CAO ; Hualei WANG ; Pei HUANG
Chinese Journal of Veterinary Science 2024;44(8):1585-1592
African swine fever(ASF),caused by the African swine fever virus(ASFV),is a highly contagious infectious disease of pigs.This disease has been spread rapidly in China since 2018,po-sing a huge threat to China's pig farming industry.To rapid detect the ASFV,a loop-mediated iso-thermal amplification(LAMP)combined with the disposable nucleic acid visualization test strip was established for visual detection of the nucleic acid of ASFV B646L gene.The method was easy to operate without special instruments and equipment,while it effectively avoided the disadvantage of false positives caused by aerosol contamination.The method was able to detect 1.16 copies/μL of the recombinant plasmid in 50 min at 65 ℃.In addition,the method was specific with no cross-re-action with classical swine fever virus,porcine reproductive and respiratory syndrome virus,por-cine parvovirus,transmissible gastroenteritis virus.The results in this study provides a rapid,con-venient,sensitive and reliable method for early diagnosis and screening for ASFV suspected infec-tion cases.
2.A visual RT-LAMP-VF method for detection of Zika virus nucleic acid
Yumeng SONG ; Pei HUANG ; Hongli JIN ; Cuicui JIAO ; Yujie BAI ; Mengyao ZHANG ; Zhiyuan GONG ; Yuanyuan LI ; Haili ZHANG ; Hualei WANG
Chinese Journal of Veterinary Science 2024;44(8):1698-1703
To establish a simple,convenient,sensitive,and specific method for rapid detection of Zi-ka virus(ZIKV),the whole genome sequences of ZIKV isolated from different times and regions were analyzed.The specific primers and probes were designed based on the screened target se-quences located in the conserved region of the ZIKV NS5 gene.By combining RT-LAMP isother-mal amplification technology and immunochromatography technology,a reverse transcription loop mediated isothermal amplification nucleic acid and flow visualization strip(RT-LAMP-VF)detec-tion method for ZIKV was established.The results showed that the method had good specificity and sensitivity.When the ratio of inner,outer,and ring primers(FIP∶LF∶F3)was 4∶2∶1,the detection method can specifically detect 102 copies/pL RNA transcripts or 2.15 pfu ZIKV at 61 ℃for 45 minutes,with no cross reaction with other flaviviruses such as Japanese encephalitis virus and classical swine fever virus.Other RNAs in blood tissue samples did not affect the sensitivity and specificity of RT-LAMP-VF,indicating that the method can be applied to clinical practice.The ZIKV RT-LAMP-VF detection method established in this study is easy to perform and does not require special instruments and equipment.It is particularly suitable for the rapid detection of ZIKV in grassroots units,providing technical support and material support for the establishment of on-site rapid detection and early warning and prediction systems for ZIKV disease.
3.Expression of severe fever with thrombocytopenia syndrome virus Gn-D Ⅲ-Ⅲ and development of indirect ELISA for antibody detection
Mengyao ZHANG ; Tianlai LIANG ; Feihu YAN ; Tao CHEN ; Cuicui JIAO ; Hongli JIN ; Jiaoyan LUAN ; Xiao WU ; Pei HUANG ; Haili ZHANG ; Qin NING ; Hualei WANG ; Yuanyuan LI
Chinese Journal of Veterinary Science 2024;44(8):1704-1712
The PCR-amplified severe fever with thrombocytopenia syndrome virus(SFTSV)Gn-DⅢ-Ⅲ gene was inserted into the pET-30a(+)prokaryotic expression vector to generate the re-combinant plasmid pET-SFTSV-Gn-D Ⅲ-Ⅲ.The plasmid was transformed into E.coli BL21(DE3)for Gn-DⅢ-m protein expression and the expression conditions were optimized.The Gn-DⅢ-Ⅲ protein purified with Ni-NTA column affinity chromatography was applied as the captured antigen to establish an indirect ELISA method for the detection of SFTSV antibody.The results demonstrated that the recombinant plasmid pET-SFTSV-Gn-D Ⅲ-Ⅲ was successfully constructed as identified by PCR and sequencing.The recombinant protein SFTSV Gn-D m-Ⅲ was soluble ex-pression in E.coli under the optimal induction conditions of 0.4 mmol/L IPTG at 25 ℃ for 4 h,and the protein purity was 91.77%after purification by Ni-NTA column.The optimal reaction con-ditions for the indirect ELISA of SFTSV antibody were as follows:coating antigen concentration(5 μg/mL),primary antibody(incubation at 37 ℃ for 1.5 h),and secondary antibody(diluted 1:10 000 and incubated at 37 ℃ for 1 h).The established method had no cross-reactivity with Rift Valley fever virus(RVFV),Ebola virus(EBOV),and tick-borne encephalitis virus(TBEV)posi-tive sera.The method had a high sensitivity,with P/N>2.1 for SFTSV-positive sera diluted to 81920.Coefficients of variation for intra-and inter-batch reactions were less than 10%.Detection of four SFTSV-infected human clinical serum samples showed the serum samples from patients in re-mission were tested as positive(P/N>2.1),while serum samples from patients with multiple or-gan failure were detected as negative(P/N<2.1).The results indicated that the SFTSV Gn-D Ⅲ-Ⅲ protein was successfully expressed and purified,and it was used as the coating protein to estab-lish an indirect ELISA assay for SFTSV antibody,which possesses good specificity,sensitivity and reproducibility.This method might be applied to detect human SFTSV clinical serum samples.
4.Famine exposure in early life and type 2 diabetes in adulthood: findings from prospective studies in China
Feng NING ; Jing ZHAO ; Lei ZHANG ; Weijing WANG ; Xiaohui SUN ; Xin SONG ; Yanlei ZHANG ; Hualei XIN ; Weiguo GAO ; Ruqin GAO ; Dongfeng ZHANG ; Zengchang PANG
Nutrition Research and Practice 2023;17(4):780-788
BACKGROUND/OBJECTIVES:
This study examined the relationship between famine exposure in early life and the risk of type 2 diabetes in adulthood during the 1959–1961 Chinese Famine.
SUBJECTS/METHODS:
A total of 3,418 individuals aged 35–74 years free of diabetes from two studies in 2006 and 2009 were followed up prospectively in 2009 and 2012, respectively. Famine exposure was classified as unexposed (individuals born in 1962–1978), fetal exposed (individuals born in 1959–1961), child exposed (individuals born in 1949–1958), and adolescent/adult exposed (born in 1931–1948). A logistic regression model was used to assess the relationship between famine exposure and diabetes after adjustment for potential covariates.
RESULTS:
During a three-year follow-up, the age-adjusted incidence rates of type 2 diabetes were 5.7%, 14.5%, 12.7%, and 17.8% in unexposed, fetal-exposed, child-exposed, and adolescent/adult-exposed groups, respectively (P < 0.01). Relative to the unexposed group, the relative risks (95% confidence interval) for diabetes were 2.15 (1.29–3.60), 1.53 (0.93– 2.51), and 1.65 (0.75–3.63) in the fetal-exposed, child-exposed, and adolescent/adult-exposed groups, after controlling for potential covariates. The interactions between famine exposure and obesity, education level, and family history of diabetes were not observed, except for the urbanization type. Individuals living in rural areas with fetal and childhood famine exposure were at a higher risk of type 2 diabetes, with relative risks of 8.79 (1.82–42.54) and 2.33 (1.17–4.65), respectively.
CONCLUSIONS
These findings indicate that famine exposure in early life is an independent predictor of type 2 diabetes, particularly in women. Early identification and intervention may help prevent diabetes in later life.
5.Application of multimodal low opioid combined with saphenous nerve block analgesia in elderly patients undergoing knee arthroplasty
Liyuan JIAO ; Ziye JING ; Hualei YAN ; Wenjie ZHANG ; Shouyuan TIAN
Chinese Journal of Geriatrics 2023;42(1):67-72
Objective:To investigate the clinical application of multimodal low-opioid combined with saphenous nerve block analgesia in elderly patients undergoing knee arthroplasty.Methods:This study is a randomized controlled study.A total of 60 elderly patients who underwent elective knee arthroplasty in the Department of Orthopedics, the First Hospital of Shanxi Medical University from January 2021 to December 2021 were selected and divided into 2 groups by numerical randomization: mode low opioid analgesia regimen group(observation group)and traditional analgesia regimen group(control group), 30 cases in each group.Observation group: (1)Preemptive analgesia: Oral celecoxib 200 mg, qd.from 3 days before surgery, the mini-mental state examination(MMSE)score was used to evaluate the cognitive function; (2)Intraoperative analgesia: After the prosthesis was installed, choose to inject analgesics around the knee joint(ropivacaine 200 mg, morphine 5 mg, epinephrine 0.25 mg, dexamethasone 5 mg/100 ml normal saline, also known as "cocktail" solution); (3)Postoperative analgesia: After the operation, continuous saphenous nerve block(0.2% ropivacaine, 2 ml/h)was performed under ultrasound guidance, and the dose of nerve block was adjusted according to the degree of rehabilitation training.Control group: no special treatment before and during the operation, traditional postoperative patient-controlled intravenous analgesia(PCIA)was used after the operation, the formula WAs as follows: sufentanil(2 μg/kg)+ flurbiprofen axetil(200-300 mg)+ Tropisetron(5-10 mg). The numerical rating scale(NRS)scores were observed and recorded 1 day before surgery, 6, 12, 24, and 48 hours after surgery; the time of the first active straight leg raising after surgery, the time of landing; the range of motion(ROM)of the knee joint 3, 7, and 14 days after surgery; the American Hospital for Special Surgery Knee Score(HSS)at 14 days, 1, 3, and 6 months after surgery; serum substance P(SP)and interleukin-6(IL-6)concentrations 1 day before surgery and 48 hours after surgery, and adverse events were recorded.Results:Compared with the control group, the patients in the observation group had lower NRS scores at 6, 12, 24, and 48 hours after the operation( t=27.705, 27.532, 21.739, 25.780, all P<0.05); the first active straight leg raising time and the time of landing earlier after the operation, and the knee joint at 3, 7, and 14 days after the operation, the range of motion(ROM)was better( t=35.496, 43.716, 3.766, 5.216, 6.009, all P<0.05). And the American hospital for special surgery knee score(HSS)was higher at 14 days, 1 month, and 3 months after surgery( t=19.247, 32.337, 22.651, all P<0.05), but there was no significant difference at 6 months after surgery.Simultaneously, the serum SP and IL-6 concentrations in the observation group 48 h after the operation were (431.0±11.3)ng/L and(11.9±2.7)ng/L, respectively.Compared with the control group(442.5±15.6)ng/L, (14.4±2.9)ng/L( t=5.362, 4.144, both P<0.05). Compared with the control group, which were lower than those in the incidence of postoperative nausea and vomiting was lower, the length of hospital stay was shorter in the observation group( χ2=4.630, t=3.311, P=0.031, 0.002), and the other indicators had no statistical differences(all P>0.05). Conclusions:Multimodal low-opioid combined with saphenous nerve block analgesia can significantly reduce perioperative pain in elderly patients undergoing knee arthroplasty, improve early postoperative mobility, and speed up postoperative functional recovery.
6.Clinical efficacy of dose escalation in 3-dimensional radiotherapy for patients with esophageal squamous cell carcinoma-multicenter retrospective analysis (3JECROG R-03)
Jingjing ZHAO ; Wencheng ZHANG ; Hualei ZHANG ; Weiming HAN ; Xin WANG ; Chen LI ; Junqiang CHEN ; Xiaomin WANG ; Yidian ZHAO ; Xueying QIAO ; Zhiguo ZHOU ; Chun HAN ; Shuchai ZHU ; Wenbin SHEN ; Lan WANG ; Xiaolin GE ; Xinchen SUN ; Kaixian ZHANG ; Miaomiao HU ; Ling LI ; Chongli HAO ; Gaofeng LI ; Yonggang XU ; Yadi WANG ; Na LU ; Miaoling LIU ; Shuai QIE ; Zefen XIAO ; Qingsong PANG ; Ping WANG
Chinese Journal of Radiation Oncology 2020;29(11):941-947
Objective:To evaluate the effect of definitive radiotherapy with different doses on overall survival (OS) and identify the prognostic factors of patients with non-metastatic esophageal squamous cell carcinoma (ESCC).Methods:Clinical data of 2 344 ESCC patients treated with definitive radiotherapy (RT) alone or chemoradiotherapy from 2002 to 2016 in 10 hospitals were collected and analyzed retrospectively. After the propensity score matching (PSM)(1 to 2 ratio), all patients were divided into the low-dose group (equivalent dose in 2 Gy fractions, EQD 2Gy<60 Gy; n=303) and high-dose group (EQD 2Gy≥60 Gy; n=606) based on the dose of radiation. Survival analysis was conducted by Kaplan- Meier method. Multivariate prognostic analysis was performed by Cox′s regression model. Results:The median follow-up time was 59.6 months. After the PSM, the 1-, 3- and 5-year overall survival (OS) rate was 66.5%, 34.7%, 27.2% in the low-dose group, 72.9%, 41.7% and 34.7% in the high-dose group, respectively ( P=0.018). The 1-, 3-and 5-year progression-free survival rate was 52.2%, 27.2%, 23.1% in the low-dose group, 58.3%, 38.1% and 33.9% in the high-dose group, respectively ( P=0.001). The outcomes of univariate analysis indicated that cervical/upper esophagus location, early (stage Ⅱ) AJCC clinical stage, node negative status, tumor length ≤5 cm, receiving intensity-modulated radiation therapy (IMRT), receiving concurrent chemotherapy and EQD 2Gy≥60 Gy were closely associated with better OS (all P<0.05). Multivariable analysis demonstrated that tumor location, regional lymph node metastasis, concurrent chemotherapy and EQD 2Gy were the independent prognostic factors for OS (all P<0.05). Conclusion:Three-dimensional conformal or IMRT with EQD 2Gy≥60 Gy yields favorable survival outcomes for patients with locally advanced ESCC.
7.Analysis on death mortality among children ≤5 years old in Jiaxing city from 2013 to 2017
Hongmei TANG ; Xiaoyan SUN ; Dinghua CHEN ; Hualei YIN ; Huanhuan HUANG ; Qinhui WU ; Yanqing YANG ; Yan ZHANG
Journal of Chinese Physician 2018;20(6):893-896
Objective To analyze the death status of children ≤ 5 years old in Jiaxing City from 2013 to 2017,and to provide scientific and effective measures of decreasing the children mortality and ensuring the health of children.Methods Death surveillance data of children ≤5 years old was obtained from monitoring network covering the whole city of maternal and child health information systems and the relevant maternal and child health statistics.The mortality rate,the sequence and changing trend of main death causes,prenatal health services of children ≤5 years old in Jiaxing City from 2013 to 2017were analyzed.Results From 2013 to 2017,there were 2 788 cases death of children ≤5 years old in Jiaxing City.The neonatal mortality rate (NMR),infant mortality rate (IMR),≤5 years old children mortality rate(U5MR) in Jiaxing City were 2.33‰,3.52‰ and 4.83 ‰,respectively.The mortality rates of all ages showed a declining trend year by year (x2NMR =19.641,P<0.001;x2IMR =31.705,P <0.001;x2U5MR =48.294,P <0.001).There were significant differences in the mortality between the lower local and the migrant children (x22013 =26.16,x22014 =18.45,x22015 =27.72,x22016 =14.49,x22017=24.19;P<0.001).The top two causes of death in children ≤ 5 years old were drowning,premature birth and low birth weight.The top two causes of death in infants and newborns were premature birth,low birth weight and other congenital anomalies.In the recent five years,49.60% of children ≤5 years old died in the hospital.At the same time,the proportion of deaths on the way and at home was 15.64% and 34.76% respectively.Although there was a gap in the level of prenatal health services between local and mobile children,the gap has been declining year by year.Conclusions The key measures to decrease the mortality rate of children under 5 years old in Jiaxing City include strengthening the management of pregnancy care and high-risk pregnancy;paying attention to tertiary prevention and doing a good job in monitoring birth defects such as prenatal screening;enhancing safety education and child supervision for parents of children in 1-4 years old;reinforcing the construction of maternal and child health professional team and the technical training of newborn resuscitation techniques such as neonatal asphyxia recovery technology;intensifying the health-care and management of migrant children and improving the ability of parents to utilize child medical service.
8. Evaluation on occupational exposure to whole body vibration in workers of a hydroelectric power station
Tao CHEN ; Huafei ZHANG ; Chuanbao YI ; Hongyan LI ; Hualei XU ; Lijun ZHAO ; Chengxun SUN ; Hansheng LIN ; Bin XIAO ; Maosheng YAN
China Occupational Medicine 2018;45(02):210-213
OBJECTIVE: To examine the level of occupational exposure to whole body vibration( WBV) in a hydroelectric power station,and evaluate its effects on workers' health. METHODS: A hydroelectric power station in Hangzhou City was chosen as study subject by using judgment sampling method. The level of WBV exposure to workers and high vibration level areas were measured,and the vibration frequency was analyzed. The comfort and allowing duration of occupational exposure in WBV exposed workers was evaluated. RESULTS: The median root-mean-square of accelerometer acceleration of WBV exposure in generator floor,turbines-floor,middle-floor,hydraulic floor of the hydroelectric power station were 0. 894,0. 995,1. 095 and 0. 904 m/s~2,respectively. The central frequency of WBV in this station was 1. 25 Hz. Among the 105 monitoring spots,8. 6% of WBV exposed workers of the spots did not perceive or feel uncomfortable,but 91. 4%of them felt uncomfortable or very uncomfortable. Only 12. 4% of the WBV exposed workers tolerated more than 8 hours of WBV occupational exposure. CONCLUSION: There is an occupational hazard of WBV in this hydroelectric power station.The low frequency vibration with central frequency below 1. 25 Hz is the key control point of the WBV of this hydroelectric power station.
9.Effect of atrial fibrillation model by high thyroxine on the electrophysiological changes in left atrium
Jialin ZHENG ; Tao GUO ; Xinjin ZHANG ; Siming TAO ; Hualei DAI ; Jianmei LI
Chinese Journal of Biochemical Pharmaceutics 2015;37(4):48-50,54
Objective To investigate about establishment the animal model of atrial fibrillation(AF) by high thyroxine and electrophysiological study of left atrium.Methods 49 rabbits were randomly divided into three groups, control group (10, injection of saline), withdrawal group (20, injection of levo-thyroxine 50μg/kg, change to inject isodose saline after two months), continuous dosing group (19, injection of levo-thyroxine 50μg/kg everyday).the data of left atrium effective refractory period(AERP), conduction velocity(CV), wavelength(WL) and AF induced ratio were collected after four months.Results The withdrawal group and continuous dosing group AERP200, AERP150 were more shorter than the control group after two months(P<0.05), The continuous dosing group AERP200, AERP150 was shorter significantly than withdrawal group and control group after four months ( P<0.01 ).The withdrawal group and continuous dosing group CV were slower than control group after two months ( P<0.05 ).The continuous dosing group CV was slower significantly than withdrawal group and control group after four months ( P<0.01 ).The withdrawal group and continuous dosing group WL were shorter than control group after two months(P<0.05), The continuous dosing group WL was shorter significantly than withdrawal group and control group after four months (P<0.01).The AF induced ratio in the continuous dosing group increased significantly(P<0.01). After four months, but the withdrawal group decreased, the control group did not induce AF.Conclusion It's feasible to establish the rabbit model of AF by high thyroxine, with left atrium electrophysiological changes, which provides animal model for further to study the pathogenesis of AF cause of hyperthyroidism.
10.Comparative analysis of whole mount processing and conventional sampling of radical prostatectomy specimens
Kun CHANG ; Xiaoqun YANG ; Chaofu WANG ; Hualei GAN ; Aihua ZHENG ; Jun YANG ; Bo DAI ; Yuanyuan QU ; Hailiang ZHANG ; Guohai SHI ; Yao ZHU ; Dingwei YE
China Oncology 2014;(11):824-829
Background and purpose:To perform whole mount technique in the diagnosis of the prostate cancer could provide orientation to the specimen. Whole mount technique has great value in pathologic diagnosis and morphological research. However, limited by the specimen-making technique, shortage of equipment and heavy workload, this technique has not been generally accepted in China. The aim of this study was to evaluate the signiifcance of whole mount technique in the diagnosis of the prostate cancer by comparing the clinical and pathological variables between whole mount patients and conventional ones after radical prostatectomy (RP).Methods:A total number of 229 patients’ whole mount RP specimens were recruited in the study from Dec. 2012 to Feb. 2014. The control group included 393 patients’ specimens which underwent conventional sampling from Jan. 2010 to Jun. 2012. We compared the clinical and pathological variables between the groups, including age, preoperative PSA level, methods of diagnosis, preliminary diagnostic Gleason score, clinical T stage, postoperative Gleason score, pathological T stage, positive surgical margin, extraprostatic extension, seminal vesicle invasion and pelvic lymph node metastasis.Results:Two groups shared similar preoperative parameters. Also there was no signiifcant difference between the whole mount and the conventional sampling groups in postoperative Gleason score, pathological T stage, extraprostatic extension and pelvic lymph node metastasis. However, positive surgical margin and seminal vesicle invasion rates were much higher in the whole mount group than the control one and both of the differences reached statistical signiifcance (26.2%vs 17.6%, 23.1%vs 17.0%;P=0.010, 0.025)Conclusion:After compared the clinical and pathological variables, we could conclude that whole mount technique has prevalence in the diagnosis of the positive surgical margin and seminal vesicle invasion compared with the conventional sampling technique. Thus, whole mount technique should be strongly recommended in the diagnosis of prostate cancer.

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