1.Construction of stable BHK-21 cell lines overexpressing APN of different species and the susceptibility to different coronaviruses
Dan WANG ; Hengjie ZHANG ; Yuyang TIAN ; Xiaohan HOU ; Zeao CHEN ; Ying HU ; Wenchao ZHANG ; Jianle REN ; Ying WANG ; Yujun ZHAO ; Ding ZHANG ; Bo YANG ; Wenxia TIAN ; Sheng NIU
Chinese Journal of Veterinary Science 2025;45(10):2095-2101
This study aims to establish BHK-21 stable cell lines expressing APN from four species(human,pig,dog,and cat),the APN fragments were amplified from pEGFP-C1-APN plasmids of the four species stored in the laboratory to generate the recombinant plasmids pcDNA4.0-APN.Af-ter the recombinant plasmids were transfected into BHK-21 cells,the stable BHK-21 cell lines ex-pressing the APNs were selected by two rounds of limited dilution.The constructed BHK-21 cell lines were identified by indirect immunofluorescence assay(IFA),and their susceptibility to PD-CoV and TGEV was tested for these four cell lines.Virus infection experiments revealed that PD-CoV infected cells expressing human,pig,and dog APNs,while it did not infect cells expressing cat APN.Simultaneously,TGEV infected cells expressing pig,dog,and cat APNs,but did not infect cells expressing human APN.The results suggest that the risk of cross-species infection for different coronaviruses and the established cell line can be used effectively to evaluate the virus in-fection.The findings also revealed that PDCoV has the potential risk of cross-species infection of human and dog,and TGEV has the potential risk of cross-species infection of dog and cat.These results provide a basis for the prevention and control strategy of coronaviruses.
2.Epidemiological investigation and post-natal follow-up analysis of hepatitis B exposed children in Wenzhou area
Lulu PAN ; Jianle SUN ; Qian XU ; Enshu WANG
Journal of Public Health and Preventive Medicine 2025;36(2):131-134
Objective To investigate the situation of hepatitis B immune blocking and the growth and development of hepatitis B exposed children within one year after birth, and to provide evidence for formulating and optimizing measures for maternal and infant blocking in Wenzhou. Methods The hepatitis B infection status during pregnancy and birth was collected, hepatitis B immunoglobulin and vaccine were analyzed, and a follow-up on the immune blocking effect, growth and development, and the nutritional status of children exposed to hepatitis B was conducted. Statistical analysis of data was carried out through SPSS 26.0. Results In 2021, 6.07% of newborns in Wenzhou were exposed to hepatitis B, of which 28.37% were highly exposed children. The prevalence rates for males and females were 6.42% and 5.70%, respectively, and the difference was statistically significant (χ2=13.744, P<0.001). The prevalence rates in mountainous and non-mountainous counties were 6.35% and 5.94%, respectively, and the difference was statistically significant (χ2=5.72, P < 0.001). In 2021, the mother-to-child transmission rate of children exposed to hepatitis B was 0.54‰. There were no significant differences in the height, weight, hemoglobin and neuropsychological development of hepatitis B exposed children compared with non-exposed children (P < 0.001). Conclusion The number of children exposed to hepatitis B in mountainous counties is significantly higher than that in non-mountainous counties in Wenzhou, and the number of men is significantly higher than that of women. The proportion of highly exposed children is relatively high. The effect of mother-to-child blocking is good, and there is no significant difference between the growth and development of children exposed to hepatitis B and no-exposed children during follow-up.
3.Feasibility and exploration of optimal communication models for robot-assisted urological telesurgery: a multicenter, single-arm, retrospective study
Ye WANG ; Taoping SHI ; Sheng TAI ; Sunyi YE ; Yubai ZHANG ; Bingzhang QIAO ; Chenfeng WANG ; Gen CHENG ; Zhi LI ; Qing AI ; Qingbo HUANG ; Baojun WANG ; Qing YUAN ; Junnan XU ; Guojun LIU ; Yu CHEN ; Wuyi ZHAO ; Jianle MAO ; Shiwei LI ; Shuo WANG ; Dan XIA ; Wanhai XU ; Chaozhao LIANG ; Hongzhao LI ; Xin MA ; Xu ZHANG
Chinese Journal of Urology 2025;46(5):331-336
Objective:To evaluate the efficacy and feasibility of a domestically developed robotic surgical system based on fiber-optic dedicated line communication in cross-regional urological telesurgery.Methods:This was multicenter,single-arm,retrospective case series study. The data of patients who underwent urological telesurgeries using the telesurgical system between January 2023 and December 2024 were analyzed. The cohort included 59 patients from seven hospitals across China. Among the patients,47 were male(79.7%)and 12 were female(20.3%),with a median age of 63.0(56.0,68.0)years and a body mass index of(24.7 ± 3.0)kg/m 2. Surgical procedures included 32 radical prostatectomies,24 partial nephrectomies,one radical nephrectomy,one adrenalectomy,and one ureteral reconstruction. The perioperative indicators,pathological results and postoperative complications were analyzed. The network monitoring data were collected,and the perioperative data of patients,remote system monitoring data and costs were compared between the two communication modes of optical transport network(OTN)and cloud-connect network(CCN). Results:All 59 remote surgeries were successfully completed,with a mean operative time of(138.0 ± 54.0)minutes,median intraoperative blood loss of 50.0(30.0,100.0)ml and a postoperative hospital stay of 5.0(4.0,6.0)days. No cases required reoperation,Clavien-Dindo grade ≥3 complications,or readmission. The geographical distance between the primary and remote surgical sites ranged from 450 to 2 800 km. Network monitoring revealed increased bidirectional latency with distance increasing:the shortest latency time(Hefei-Hangzhou,450 km)was(16.59 ± 0.80)ms,while the longest(Harbin-Hangzhou,2 200 km)latency time was(53.31 ± 0.31)ms. Average frame loss per procedure was 0?1.27 frames. The results of subgroup analysis comparing OTN and CCN communication modes showed no significant differences in operative time[(130.7 ± 70.5)minutes vs.(142.1 ± 42.9)minutes, P = 0.442],postoperative hospitalization[6.0(4.0,8.0)d vs. 5.0(4.0,6.0)d, P = 0.581],or readmission rates(0 vs. 0). However,CCN demonstrated significant cost advantages with 500 RMB per operation vs. 3 000 RMB per operation for OTN. Conclusions:Urological telesurgery using fiber-optic communication is feasible. The CCN mode,with its cost-effectiveness,excellent usability,and multi-point interconnection flexibility,is currently the preferred communication model for telesurgical applications.
4.Construction of stable BHK-21 cell lines overexpressing APN of different species and the susceptibility to different coronaviruses
Dan WANG ; Hengjie ZHANG ; Yuyang TIAN ; Xiaohan HOU ; Zeao CHEN ; Ying HU ; Wenchao ZHANG ; Jianle REN ; Ying WANG ; Yujun ZHAO ; Ding ZHANG ; Bo YANG ; Wenxia TIAN ; Sheng NIU
Chinese Journal of Veterinary Science 2025;45(10):2095-2101
This study aims to establish BHK-21 stable cell lines expressing APN from four species(human,pig,dog,and cat),the APN fragments were amplified from pEGFP-C1-APN plasmids of the four species stored in the laboratory to generate the recombinant plasmids pcDNA4.0-APN.Af-ter the recombinant plasmids were transfected into BHK-21 cells,the stable BHK-21 cell lines ex-pressing the APNs were selected by two rounds of limited dilution.The constructed BHK-21 cell lines were identified by indirect immunofluorescence assay(IFA),and their susceptibility to PD-CoV and TGEV was tested for these four cell lines.Virus infection experiments revealed that PD-CoV infected cells expressing human,pig,and dog APNs,while it did not infect cells expressing cat APN.Simultaneously,TGEV infected cells expressing pig,dog,and cat APNs,but did not infect cells expressing human APN.The results suggest that the risk of cross-species infection for different coronaviruses and the established cell line can be used effectively to evaluate the virus in-fection.The findings also revealed that PDCoV has the potential risk of cross-species infection of human and dog,and TGEV has the potential risk of cross-species infection of dog and cat.These results provide a basis for the prevention and control strategy of coronaviruses.
5.Feasibility and exploration of optimal communication models for robot-assisted urological telesurgery: a multicenter, single-arm, retrospective study
Ye WANG ; Taoping SHI ; Sheng TAI ; Sunyi YE ; Yubai ZHANG ; Bingzhang QIAO ; Chenfeng WANG ; Gen CHENG ; Zhi LI ; Qing AI ; Qingbo HUANG ; Baojun WANG ; Qing YUAN ; Junnan XU ; Guojun LIU ; Yu CHEN ; Wuyi ZHAO ; Jianle MAO ; Shiwei LI ; Shuo WANG ; Dan XIA ; Wanhai XU ; Chaozhao LIANG ; Hongzhao LI ; Xin MA ; Xu ZHANG
Chinese Journal of Urology 2025;46(5):331-336
Objective:To evaluate the efficacy and feasibility of a domestically developed robotic surgical system based on fiber-optic dedicated line communication in cross-regional urological telesurgery.Methods:This was multicenter,single-arm,retrospective case series study. The data of patients who underwent urological telesurgeries using the telesurgical system between January 2023 and December 2024 were analyzed. The cohort included 59 patients from seven hospitals across China. Among the patients,47 were male(79.7%)and 12 were female(20.3%),with a median age of 63.0(56.0,68.0)years and a body mass index of(24.7 ± 3.0)kg/m 2. Surgical procedures included 32 radical prostatectomies,24 partial nephrectomies,one radical nephrectomy,one adrenalectomy,and one ureteral reconstruction. The perioperative indicators,pathological results and postoperative complications were analyzed. The network monitoring data were collected,and the perioperative data of patients,remote system monitoring data and costs were compared between the two communication modes of optical transport network(OTN)and cloud-connect network(CCN). Results:All 59 remote surgeries were successfully completed,with a mean operative time of(138.0 ± 54.0)minutes,median intraoperative blood loss of 50.0(30.0,100.0)ml and a postoperative hospital stay of 5.0(4.0,6.0)days. No cases required reoperation,Clavien-Dindo grade ≥3 complications,or readmission. The geographical distance between the primary and remote surgical sites ranged from 450 to 2 800 km. Network monitoring revealed increased bidirectional latency with distance increasing:the shortest latency time(Hefei-Hangzhou,450 km)was(16.59 ± 0.80)ms,while the longest(Harbin-Hangzhou,2 200 km)latency time was(53.31 ± 0.31)ms. Average frame loss per procedure was 0?1.27 frames. The results of subgroup analysis comparing OTN and CCN communication modes showed no significant differences in operative time[(130.7 ± 70.5)minutes vs.(142.1 ± 42.9)minutes, P = 0.442],postoperative hospitalization[6.0(4.0,8.0)d vs. 5.0(4.0,6.0)d, P = 0.581],or readmission rates(0 vs. 0). However,CCN demonstrated significant cost advantages with 500 RMB per operation vs. 3 000 RMB per operation for OTN. Conclusions:Urological telesurgery using fiber-optic communication is feasible. The CCN mode,with its cost-effectiveness,excellent usability,and multi-point interconnection flexibility,is currently the preferred communication model for telesurgical applications.
6.Analysis of treatment strategies for donor-derived infection: a report of 486 cases
Shengli CAO ; Xiaoyi SHI ; Peihao WEN ; Jianle HAN ; Changan WANG ; Wenzhi GUO ; Shuijun ZHANG
Chinese Journal of Organ Transplantation 2022;43(3):135-140
Objective:To explore the microbiological characteristics of donor blood culture and donor liver perfusion culture and summarize the clinical experiences to provide basic rationales for preventing donor-derived infections.Methods:From August 1, 2018 to November 26, 2018 and November 27, 2018 to December 31, 2020 at First Affiliated Hospital, Zhengzhou University, culture results of donor blood and donor liver perfusate were retrospectively reviewed.According to whether or not donor liver was obtained without breaking diaphragm, removing gallbladder intraoperatively and flushing bile through cystic duct, two stages were assigned: before and after improvement measures of liver donor, i.e.August 1, 2018 to November 26, 2018 and November 27, 2018 to December 31, 2020.The culture results of donor blood samples and donor liver perfusion fluid samples in two stages of liver transplantation were statistically analyzed and infection preventing measures during donor liver maintenance and obtaining donor liver examined.Results:A total of 486 cases of blood culture from potential donors and 478 cases of liver perfusion culture were analyzed.The results showed that the incidence of blood culture infection was 4.5% and 4.3% before and after improvement measures( χ2=0.008; P=0.927)while the incidence of perfusion fluid infection was 56.8% and 46.2%( χ2=4.569; P=0.031); Klebsiella pneumoniae was a major pathogen cultured in perfusion solution before improvement measures and Staphylococcus epidermidis after improvement measures. Conclusions:Before organ donation, infection screening and prevention of potential donors and corresponding measures during donor liver acquisition can reduce donor source infection and effectively lower the mortality of recipients.
7.Correlation between serum level of miRNA-106a expression with clinicopathological characteristics and prognosis of patients with renal cell carcinoma.
Qingyan YANG ; Junyi LIU ; Yalin LIANG ; Changan WANG ; Jianle HAN ; Litao ZHU ; Shengping YUAN ; Qiang SUN ; Hongsen ZHANG
Chinese Journal of Medical Genetics 2021;38(7):652-655
OBJECTIVE:
To analyze the expression of microRNA-106a(miR-106a) in renal cell carcinoma (RCC) and its correlation with clinicopathological characteristics and prognosis of patients.
METHODS:
Serum samples of 64 patients with newly diagnosed RCC were collected as the study group, and serum samples of 40 healthy individuals were used as the control group. Real-time fluorescence quantitative PCR was used to determine the expression level of miR-106a in each group. The correlation between miR-106a expression and clinicopathological characteristics of the patients was studied with single factor analysis and multiple Logistic regression model. Kaplan-Meier survival curve was used to analyze its correlation with the prognosis of patients.
RESULTS:
Before surgery, compared with the control group (1.17± 0.58), RCC patients with high- (9.15± 0.96) and low-expression(3.45± 0.37) had increased expression of miR-106a. Postoperatively, the expression level of miR-106a in both groups of patients decreased to 1.53± 0.18 and 1.75± 0.21, respectively. The area under the curve (AUC) of the diagnostic value of serum miR-106a for RCC was 0.782 (95% CI: 0.661-0.902). With an optimal cutoff value of 0.531, the sensitivity was 78.10% and the specificity was 75.00%. Serum miR-106a level of RCC patients with TNM stage T3 or T4, clinical stage II or III, lymph node metastasis, and recurrence were significantly increased. The high expression of serum miR-106a in RCC patients has an independent relationship with the tumor TNM stage and lymph node metastasis. Of the 64 follow-up patients, 4 were lost and 30 had died. Among them, the median survival time of patients in the miR-106a high expression group was 30 months, which was significantly shorter than that of the low expression group (52 months).
CONCLUSION
The serum level of miR-106a is elevated in RCC patients, and may be used as a molecular marker for the diagnosis of RCC. High serum expression of miR-106a is an independent predictor for tumor TNM stage and lymph node metastasis, as well as an independent predictor for poor prognosis of RCC patients.
Biomarkers, Tumor/genetics*
;
Carcinoma, Renal Cell/genetics*
;
Gene Expression Regulation, Neoplastic
;
Humans
;
Kidney Neoplasms/genetics*
;
MicroRNAs/genetics*
;
Neoplasm Recurrence, Local
;
Prognosis
8.Research progress in circular RNAs in spine and spinal cord diseases
Jianle WANG ; Jun GAO ; Yizhen HUANG ; Gang LIU ; Weiyu NI ; Jianjun MA ; Xiangqian FANG ; Shuying SHEN ; Shunwu FAN
Chinese Journal of Orthopaedics 2020;40(11):743-748
Circular RNA (circRNA) is a kind of RNA with a circular structure. The unique structure of circRNA endows it with various cell biological functions and characteristics. It has become a research hotspot recently. CircRNA can play a role via mechanisms, such as microRNA (miRNA) sponge, RNA binding protein, peptide translation and regulation of gene transcription. CircRNA was found to be associated with disc degeneration, spinal cord injury, scoliosis, and facet arthritis. Some techniques, including bioinformatics and molecular biology techniques, microarray and high-throughput sequencing, can be used to predict and to discover disease-related circRNA, aiming to evaluate whether circRNA can be used as a molecular biomarker for spinal and spinal cord diseases. Based on the current role of circRNA, the corresponding therapeutic strategies have been carried out in experimental animals, which can provide theoretical basis for gene therapy. At present, the researches in circRNA for spinal and spinal cord diseases are still insufficient compared with those in other fields. Currently, the main direction focuses on the miRNA sponge mechanism of circRNA. Due to the variety of diseases in spinal surgery, the research progress of circRNA is also varied. In addition, the development of microarray and high-throughput sequencing technology have greatly promoted the researches in circRNA. The availability of public database is of great significance in the study. The present review summarized the current researches status of circRNA in spinal and spinal cord diseases, aiming to deepen understanding of circRNA in spinal and spinal cord diseases.
9. Association between prion infection and Ryanodine receptor 2 in scrapie-263 K infected hamster
Yao LI ; Jianle LI ; Kang XIAO ; Yue MA ; Liping GAO ; Jing WANG ; Zhen ZHEN ; Yanjun GUO ; Qi SHI
Chinese Journal of Experimental and Clinical Virology 2018;32(5):474-478
Objective:
To classify the expression of RyR2 in the brain of prion infected hamsters.
Methods:
Immunohistochemical assays was used to verify that the location of RyR2 in the brain slices of hamster. Assays of the brain samples of intracranial inoculation of scrapie infected hamster (agent 263 K) and normal hamster were tested to evaluate the RyR2 expression by Western Blotting. Immunofluorescent assays were used to verify the co-location between RyR2 and PrP protein.
Results:
RyR2 mainly located in cortex and Purkinje cells with parts of which are distributed in thalamus, hippocampus and olfactory bulb. The expression of RyR2 significantly decreased in the 263 K infected hamster at terminal stage. The Immunofluorescence tests showed that RyR2 was colocalized with PrP protein.
Conclusions
The experimental data showed that RyR2 may play a crucial role in prion disease, which might be closely linked to the cognition impairment and neuron loss. The relation between RyR2 and prion disease still needs further research.
10.Mortality of kidney transplantation recipients with type 2 diabetes: a retrospective cohort study
Jinrui LIU ; Chang'an WANG ; Jinghua ZHANG ; Jianle HAN
Chinese Journal of Organ Transplantation 2018;39(11):671-675
Objective To compare the difference of all-cause mortality between diabetic and non-diabetic transplant recipients.Methods This population cohort study included all primary kidneyonly transplant recipients with data integrity between Jan.2000 and May 2017,the deadline date of follow-up was May 2018,and median follow-up period was 5.9 (range 1.1 to 17.1) years.According to whether the recipients had diabetes,they were divided into type 2 diabetes group (DM group),and no diabetes group (non-DM group).The clinical data of the two groups including recipients and donors were collected.The all-cause mortality and difference in dead causes were analyzed.Results Mortality was higher in DM group than in non-DM group.Compared with non-DM group,the relative risk(RR) for all-cause mortality in DM group was 2.25 (95% CI:1.23-4.12).The cumulative survival rate in non-DM group was significantly higher than that in DM group(Log Rank P =0.035).The DM group aged younger than 40 years had the highest risk (RR 4.50),but there was no significant difference between the two groups at the age of ≥40 years.Compared with living donor and cadaveric donor renal transplantation,the relative mortality risk in DM group in DCD donor renal transplantation was highest (RR 2.68).The transplantation time did not change the multiple risk of death in both groups (adjusted RR 2.13).The first cause of death in two groups was infection (50% in DM group vs.53.3% in non-DM group).The difference was not statistically significant.Conclusion Kidney transplant recipients with type 2 diabetes had higher all-cause mortality than in those without diabetes.Compared with non-diabetic recipients,the age <40 years and receiving DCD for kidneys had a greater impact on the all-cause mortality of diabetic recipients.Infection was the leading cause of death in both groups.


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