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.Practice and reflection on promoting development through interdisciplinary integration in an affiliated hospital of a university
Lihui YU ; Zhong CHEN ; Lu DING ; Fang WANG ; Mingxia YU ; Zhongqiang YUE
Chinese Journal of Medical Science Research Management 2025;38(3):203-213
Objective:To summarize the theoretical basis and practical experience of the hospital where the author works, in actively promoting the interdisciplinary integration of ″Medicine+ ″, and provide reference for hospitals of the same type.Methods:Based on the Resource-Based View framework, this study analyzed the core dimension heterogeneous resources including organizational capital, physical and financial capital, and human capital through a case study of Zhongnan Hospital of Wuhan University, systematically examining its interdisciplinary research promotion strategies.Results:The hospital had created a strong interdisciplinary atmosphere. Its scientific research and innovation capabilities had been significantly enhanced. High-quality interdisciplinary academic achievements had emerged continuously. And its academic influence has continued to expand.Conclusions:Interdisciplinary integration is a systematic project that requires organized implementation and a ″multi-pronged approach″: coordinating and innovating the organizational management system, conducting comprehensive resource inventorying and ensuring sustainable provision, making overall plans for talent introduction and cultivation, and adhering to the clinical-problem-oriented approach in carrying out interdisciplinary integration.
3.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.
4.Practice and reflection on promoting development through interdisciplinary integration in an affiliated hospital of a university
Lihui YU ; Zhong CHEN ; Lu DING ; Fang WANG ; Mingxia YU ; Zhongqiang YUE
Chinese Journal of Medical Science Research Management 2025;38(3):203-213
Objective:To summarize the theoretical basis and practical experience of the hospital where the author works, in actively promoting the interdisciplinary integration of ″Medicine+ ″, and provide reference for hospitals of the same type.Methods:Based on the Resource-Based View framework, this study analyzed the core dimension heterogeneous resources including organizational capital, physical and financial capital, and human capital through a case study of Zhongnan Hospital of Wuhan University, systematically examining its interdisciplinary research promotion strategies.Results:The hospital had created a strong interdisciplinary atmosphere. Its scientific research and innovation capabilities had been significantly enhanced. High-quality interdisciplinary academic achievements had emerged continuously. And its academic influence has continued to expand.Conclusions:Interdisciplinary integration is a systematic project that requires organized implementation and a ″multi-pronged approach″: coordinating and innovating the organizational management system, conducting comprehensive resource inventorying and ensuring sustainable provision, making overall plans for talent introduction and cultivation, and adhering to the clinical-problem-oriented approach in carrying out interdisciplinary integration.
5.Evidence-based clinical guideline for the diagnosis and treatment of surgical site infection in spinal trauma (version 2024)
Zhu GUO ; Chao WANG ; Hongfei XIANG ; Zhongqiang CHEN ; Liang CHEN ; Tongwei CHU ; Shucai DENG ; Jian DONG ; Xinru DU ; Shiqing FENG ; Baorong HE ; Xijing HE ; Jianzhong HU ; Yong HAI ; Qingquan KONG ; Guiqing LIANG ; Qi LIAO ; Zhongjun LIU ; Shaoyu LIU ; Baoge LIU ; Xiaoguang LIU ; Weishi LI ; Li LI ; Fang LI ; Bin LIN ; Shibao LU ; Tao NIU ; Zhenli QIAO ; Dike RUAN ; Yueming SONG ; Haipeng SI ; Jun SHU ; Zhongyi SUN ; Qing WANG ; Zili WANG ; Huan WANG ; Hongli WANG ; Yan WANG ; Xiaolin WU ; Zhanyong WU ; Jinglong YAN ; Tengbo YU ; Qiang ZHANG ; Guoqing ZHANG ; Xuesong ZHANG ; Fengdong ZHAO ; Jie ZHAO ; Zhaomin ZHENG ; Qingsan ZHU ; Dingjun HAO ; Bohua CHEN
Chinese Journal of Trauma 2024;40(12):1057-1070
Spinal surgical site infection (SSI), especially deep SSI after internal fixation is difficult in treatment, with long course of disease and poor prognosis. At present, there are many controversies in the diagnosis and treatment of spinal SSI, with unsatisfactory overall efficacy of its diagnosis and treatment. Besides, no diagnosis and treatment guideline based on evidence-based medicine has been in existence. To this end, the Spinal Infection Group of the Orthopedic Branch of the Chinese Medical Doctor Association and the Spinal Infection Group of the Spinal Surgery Branch of the Chinese Rehabilitation Medicine Association jointly organized relevant experts to formulate Evidence-based clinical guideline for the diagnosis and treatment of surgical site infection in spinal trauma ( version 2024) based on an evidence-based approach. A total of 10 recommendations were proposed on the diagnosis and treatment of spinal SSI, so as to provide a clinical reference for the diagnosis and treatment of spinal SSI.
6.Value of Two-Dimensional Transverse Section of Thalamus in Evaluating the Developmental Status of Lateral Fissure of Fetus
Zhongqiang ZHU ; Xingyun WANG ; Ling REN
Chinese Journal of Medical Imaging 2024;32(12):1262-1265,1276
Purpose To investigate the value of two-dimension thalamic horizontal transverse section in the assessment of normal fetal lateral fissure development at 20-36+6 weeks. Materials and Methods It was a prospective cross-sectional study,normal single pregnant women who were examined in the First Affiliated Hospital of Gannan Medical University from March 1,2023 to March 1,2024 were selected for fetal brain imaging via two-dimensional ultrasound imaging technology. The morphology of lateral fissure of 272 normal fetuses at 20-36+6 weeks were observed with two-dimensional transverse section of thalamus. Lateral cleft width and depth were measured at a two-dimensional thalamic horizontal cross-section and a reference range of normal fetal lateral cleft width and depth was established at 20-36+6 weeks. The repeatability of lateral crack width and depth measurements within and between observers were all verified. Results A total of 272 normal fetuses of 20-36+6 weeks were enrolled in the study. A range of reference values for lateral cleft width and depth of normal fetus at 20-36+6 weeks of normal gestation was established. The lateral fetal fissure width and depth increased linearly with gestational age. The lateral fetal fissure width (mm)=-16.92+1.18 gestational age,and the lateral fetal fissure depth (mm)=-9.77+0.86 gestational age,with good intra-and inter-observer agreement (intra-class correlation coefficient,ICC>0.850). More than 95% of the points in each Bland-Altman plot fall within the 95% consistency limit. Conclusion Two-dimensional thalamic horizontal transverse section can be used to observe the morphological changes of lateral fella in middle and late pregnancy. It is a simple and feasible method for prenatal assessment of cerebral cortex development,and can be used to guide follow-up,referral,prenatal consultation and other work.
7.Value of Two-Dimensional Transverse Section of Thalamus in Evaluating the Developmental Status of Lateral Fissure of Fetus
Zhongqiang ZHU ; Xingyun WANG ; Ling REN
Chinese Journal of Medical Imaging 2024;32(12):1262-1265,1276
Purpose To investigate the value of two-dimension thalamic horizontal transverse section in the assessment of normal fetal lateral fissure development at 20-36+6 weeks. Materials and Methods It was a prospective cross-sectional study,normal single pregnant women who were examined in the First Affiliated Hospital of Gannan Medical University from March 1,2023 to March 1,2024 were selected for fetal brain imaging via two-dimensional ultrasound imaging technology. The morphology of lateral fissure of 272 normal fetuses at 20-36+6 weeks were observed with two-dimensional transverse section of thalamus. Lateral cleft width and depth were measured at a two-dimensional thalamic horizontal cross-section and a reference range of normal fetal lateral cleft width and depth was established at 20-36+6 weeks. The repeatability of lateral crack width and depth measurements within and between observers were all verified. Results A total of 272 normal fetuses of 20-36+6 weeks were enrolled in the study. A range of reference values for lateral cleft width and depth of normal fetus at 20-36+6 weeks of normal gestation was established. The lateral fetal fissure width and depth increased linearly with gestational age. The lateral fetal fissure width (mm)=-16.92+1.18 gestational age,and the lateral fetal fissure depth (mm)=-9.77+0.86 gestational age,with good intra-and inter-observer agreement (intra-class correlation coefficient,ICC>0.850). More than 95% of the points in each Bland-Altman plot fall within the 95% consistency limit. Conclusion Two-dimensional thalamic horizontal transverse section can be used to observe the morphological changes of lateral fella in middle and late pregnancy. It is a simple and feasible method for prenatal assessment of cerebral cortex development,and can be used to guide follow-up,referral,prenatal consultation and other work.
8.Evidence-based clinical guideline for the diagnosis and treatment of surgical site infection in spinal trauma (version 2024)
Zhu GUO ; Chao WANG ; Hongfei XIANG ; Zhongqiang CHEN ; Liang CHEN ; Tongwei CHU ; Shucai DENG ; Jian DONG ; Xinru DU ; Shiqing FENG ; Baorong HE ; Xijing HE ; Jianzhong HU ; Yong HAI ; Qingquan KONG ; Guiqing LIANG ; Qi LIAO ; Zhongjun LIU ; Shaoyu LIU ; Baoge LIU ; Xiaoguang LIU ; Weishi LI ; Li LI ; Fang LI ; Bin LIN ; Shibao LU ; Tao NIU ; Zhenli QIAO ; Dike RUAN ; Yueming SONG ; Haipeng SI ; Jun SHU ; Zhongyi SUN ; Qing WANG ; Zili WANG ; Huan WANG ; Hongli WANG ; Yan WANG ; Xiaolin WU ; Zhanyong WU ; Jinglong YAN ; Tengbo YU ; Qiang ZHANG ; Guoqing ZHANG ; Xuesong ZHANG ; Fengdong ZHAO ; Jie ZHAO ; Zhaomin ZHENG ; Qingsan ZHU ; Dingjun HAO ; Bohua CHEN
Chinese Journal of Trauma 2024;40(12):1057-1070
Spinal surgical site infection (SSI), especially deep SSI after internal fixation is difficult in treatment, with long course of disease and poor prognosis. At present, there are many controversies in the diagnosis and treatment of spinal SSI, with unsatisfactory overall efficacy of its diagnosis and treatment. Besides, no diagnosis and treatment guideline based on evidence-based medicine has been in existence. To this end, the Spinal Infection Group of the Orthopedic Branch of the Chinese Medical Doctor Association and the Spinal Infection Group of the Spinal Surgery Branch of the Chinese Rehabilitation Medicine Association jointly organized relevant experts to formulate Evidence-based clinical guideline for the diagnosis and treatment of surgical site infection in spinal trauma ( version 2024) based on an evidence-based approach. A total of 10 recommendations were proposed on the diagnosis and treatment of spinal SSI, so as to provide a clinical reference for the diagnosis and treatment of spinal SSI.
9.Clinical application of optical surface monitoring system in radiotherapy after radical mastectomy for left breast cancer
Bei KUANG ; Zhongqiang LIU ; Hui WANG ; Zhenming WU ; Xiqu YE ; Gengxian LIN
Chinese Journal of Radiological Health 2023;32(1):40-45
Objective To study the setup error under deep inspiration breath hold (DIBH) guided by optical surface monitoring system (OSMS) and free breathing (FB) FB1 and FB2 (without OSMS guidance, directly set up the body marker line by laser lamp) in radiotherapy after radical mastectomy for left breast cancer, and to provide a basis for individualized clinical target volume-planning target volume (CTV-PTV) expansion for the doctor in charge to delineate the target volume. Methods A total of 36 patients with left breast cancer after radical mastectomy were selected and divided into three groups, in which cone beam computed tomography (CBCT) images were taken in three states: DIBH, FB1, and FB2, respectively. CBCT and CT images were analyzed for registration; the absolute error data of linear displacement in the ventro-dorsal, cranio-caudal, and left-right directions were recorded, and the expanding margin was calculated. Results The translation errors in the ventro-dorsal, cranio-caudal, and left-right directions were (0.06 ± 0.22) cm, (0.05 ± 0.23) cm, and (0.01 ± 0.24) cm in the DIBH group, (0.07 ± 0.21) cm, (0.02 ± 0.23) cm, and (0.02 ± 0.21) cm in the FB1 group, and (0.07 ± 0.24) cm, (0.07 ± 0.34) cm, and (0.25 ± 0.09) cm in the FB2 group. The statistical results of the DIBH group and FB1 group in the ventro-dorsal, RTN, and ROLL directions were significantly different (P < 0.05). The statistical results of the FB1 group and FB2 group in the ventro-dorsal direction were significantly different. The relation of three groups in the value of margin of planning target volume was DIBH < FB1 < FB2 in the ventro-dorsal and cranio-caudal directions and FB1 < DIBH < FB2 in the left-right direction. Conclusion OSMS-guided DIBH radiotherapy in patients with left breast cancer after radical mastectomy can reduce the setup error and provide an important basis for individualized CTV-PTV expansion for the doctor in charge to delineate the target volume.
10.Effects of TRIB3 activation of Wnt/β-catenin signaling pathway on growth and proliferation of laryngeal carcinoma TU686 cells in vitro and peripheral immunosuppressive molecule expression in transplanted mice
Zhongqiang CHENG ; Chengyi JIANG ; Wei WANG ; Hualong QIANG ; Xiaodong ZHAN ; Runsheng YUAN
Chinese Journal of Immunology 2023;39(12):2595-2600
Objective:To investigate the effects of TRIB3 activation of Wnt/β-catenin signaling pathway on the growth and pro-liferation of laryngeal carcinoma TU686 cells in vitro and expression of peripheral immunosuppressive molecules in transplanted mice.Methods:Protein and RNA expressions of TRIB3 were detected in vitro cells(human immortalized epidermal cell line HaCat and laryngeal carcinoma cell line TU686)and tissues(laryngeal carcinoma and adjacent tissues),respectively.Laryngeal carcinoma TU686 cells were cultured in vitro and divided into negative control group(NC group)and TRIB3 knockdown group(sh-TRIB3 group),total protein and RNA of cells were extracted to verify the expression level of TRIB3 in two groups.After successful verifica-tion,proliferation ability of TU686 cells was detected by CCK-8,colony cloning and flow cytometry.Protein expression levels of Wnt,Cyclin-D1,C-myc,β-catenin and p-β-catenin in two groups were detected by Western blot.Correlation analysis verified the correla-tion between TRIB3 and Wnt,Cyclin-D1,C-myc,β-catenin,p-β-catenin protein expressions.TRIB3-low expressing nude mouse transplanted tumor model(TRIB3 sgRNA group)was constructed by knockdown the TRIB3 core plasmid,and a parallel control group(Control sgRNA group)was set up,tumor growth volume and weight were observed,and serum immunosuppressive molecules expres-sions was determined by ELISA.Results:Compared with HaCat cells and normal paracarcinoma tissues,TRIB3 was highly expressed in TU686 cells and laryngeal carcinoma tissues.Compared with negative control group,proliferation ability of TU686 cells was signifi-cantly inhibited after TRIB3 knockdown,and cell growth was blocked in G1/S phase.Expressions of Wnt,Cyclin-D1,C-myc andβ-catenin protein in Wnt/β-catenin signaling pathway were decreased significantly,while expression of p-β-catenin was increased significantly.TRIB3 was significantly correlated with protein expression levels of Wnt,Cyclin-D1,β-catenin and p-β-catenin.The in vivo results showed that compared with Control sgRNA group,tumor growth volume and weight of mice in TRIB3 sgRNA group were significantly decreased,and expressions of serum immunosuppressive molecules IL-4,IL-6,IL-10,TGF-β and PGE2 were signifi-cantly decreased.Conclusion:TRIB3 is highly expressed in TU686 cells,and TRIB3 can inhibit growth and proliferation of TU686 cells and transplanted tumors by activating Wnt/β-catenin-related signaling pathways,and reverse tumor immunosuppressive microen-vironment,suggesting that TRIB3 may be an effective target for laryngeal cancer.


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