1.Screening and characterization of anti-SARS-CoV-2 receptor binding domain antibodies with broad-spectrum neutralizing activity
NI Wanqi ; REN Li ; JIN Changzhong ; YANG Furong ; SHEN Yumin ; WANG Shuo ; HU Caiqin ; HAO Yanling ; LIU Ying ; ZHU Biao ; SHAO Yiming ; LI Dan ; WANG Zheng
China Tropical Medicine 2024;24(3):258-
convalescents, and to screen for broad-spectrum neutralizing antibodies against the SARS-CoV-2 RBD. Methods Using biotinylated RBD as a molecular probe, flow cytometry was employed to perform single-cell sorting of B cells from peripheral blood mononuclear cells (PBMCs) of convalescents. The obtained B cells were lysed and subjected to reverse transcription, followed by nested PCR amplification of the heavy and light chains of antibodies was conducted using random primers. The amplified products were cloned into corresponding expression vectors, and the respective matched heavy-light chain plasmids were co-transfected into 293F cells for expression. Monoclonal antibodies were then purified using Protein A column chromatography. Neutralization experiments were conducted with the wild-type (WT) pseudovirus, and antibodies with IC50<0.1 μg/mL were selected for further testing of neutralizing breadth and potency against the wild-type (WT), Beta variant (B.1.351), Delta variant (B.1.617.2), and currently prevalent pseudovirus strains (XBB, BA.5, BF.7). Results A total of 21 RBD-specific monoclonal B cells were obtained from two recovered patients, resulting in the isolation of 13 pairs of antibody light/heavy chains. Nine antibodies were successfully expressed, with P1-A1, P1-B6, and P1-B9 exhibiting IC50 values below 0.1 μg/mL against the pseudovirus of the wild-type strain (WT). Specifically, P1-B6 effectively neutralized the wild-type strain (WT), Beta variant (B.1.351), and Delta variant (B.1.617.2), with IC50 values reaching 0.01 μg/mL. P1-B9 demonstrated effective neutralization against the wild-type strain (WT), Beta variant (B.1.351), Delta variant (B.1.617.2), and Gamma variant (P.1) pseudoviruses, with IC50 values of 0.42 μg/mL, 0.63 μg/mL, 0.28 μg/mL, and 2.50 μg/mL, respectively. Additionally, P1-B6 exhibited good neutralization against BA.5 and BF.7 pseudoviruses, with IC50 values of 0.06 μg/mL and 0.09 μg/mL, respectively. Conclusions Infection with the SARS-CoV-2 WT strain can induce the generation of neutralizing antibodies with broad-spectrum activity. Generating these broadly neutralizing antibodies does not require an excessively high somatic hypermutation. The obtained antibodies can be used as candidates for SARS-CoV-2 diagnosis and prevention.
2.Clinical study of retrograde intrarenal surgery and miniaturized percutaneous nephrolithotomy in the treatment of lower pole kidney stones with a diameter <1.5 cm
Chengcun ZHU ; Fan CHENG ; Weimin YU ; Jinsong AO ; Bo WANG ; Chengjun QIU ; Dan YU ; Changzhong WANG
Chinese Journal of Urology 2023;44(1):32-36
Objective:To evaluate the clinical efficacy and safety of retrograde intrarenal surgery(RIRS) and miniaturized percutaneous nephrolithotomy(mini-PCNL) in the treatment of lower pole kidney stones with a diameter <1.5 cm.Methods:The data of 95 patients with lower pole kidney stones with a diameter <1.5 cm treated in Renmin Hospital of Wuhan University from June 2017 to October 2020 were retrospectively analyzed. According to different surgical methods, the patients were divided into RIRS group and mini-PCNL group. There were 51 cases in RIRS group and 44 cases in mini-PCNL group. There was no significant difference in age [(48.2±11.4) years vs. (46.4±14.1) years], body mass index [(21.9±2.4) kg/m 2 vs. (20.7±3.2) kg/m 2], gender [male/female: 37/14 vs. 24/20], stone CT [(746.42±164.24)HU vs. (858.62±148.72)HU], creatinine [(71.3±21.6)μmol/L vs. (63.5±20.3)μmol/L], stone location (left/right: 26/25 vs. 23/21), stone diameter [(10.5±2.1) mm vs. (12.5±2.4) mm], infundibulopelvic angle [(43.32±9.42) degrees vs. (43.82±10.34) degrees], infundibular length [(24.92±4.85)mm vs. (24.37±5.26)mm] and infundibular [(9.26±3.04)mm vs.(9.46±2.94)mm] between the two groups ( P>0.05). The operation time, stone-free rate, hospital stay and postoperative complications between the two groups were compared. Results:Compared with the mini-PCNL group, the RIRS group had significantly smaller decrease in postoperative hemoglobin [(1.53±0.92) g/L vs. (4.54±2.46) g/L, P<0.05], the postoperative hospital stay was shorter [(2.52±0.94) d vs. (4.51±1.25)d, P<0.05], and postoperative visual analogue score was lower [(2.43±0.92) vs. (3.24±0.76), P<0.05]. The operation time of the mini-PCNL group was shorter than that of the RIRS group [(42.32±13.28) min vs. (54.24±14.43)min, P<0.05]. There was no significant difference in postoperative complications [5.9% (3/51) vs. 11.4% (5/44), P>0.05], postoperative cveatinine [(71.3±21.6) μmol/L vs. (63.5±20.3) μmol/L, P>0.05], postoperative intestinal function recovery time [(25.46±10.28)h vs. (32.43±9.25)h, P>0.05] and stone-free rate [92.2% (47/51) vs. 97.7% (43/ 44), P>0.05] between the two groups. Conclusions:Both RIRS and mini-PCNL are effective and safe minimally invasive treatments for lower pole kidney stones with a diameter < 1.5 cm. RIRS has shorter operation time, less blood loss, lower pain score and faster postoperative recovery.
3.IL-17 Imbalance Promotes thePyroptosis in Immune-Mediated L
Wenfang XU ; Yanan WANG ; Changzhong JIN ; Weiyang ZHANG ; Jiangnan CHEN ; Xuefang CHEN ; Junli GAO ; Junshun GAO ; Hong WANG
Immune Network 2023;23(6):e46-
Autoimmune hepatitis (AIH) affects all age group and occurs mainly in women. Pyroptosis is a novel programmed cell death featured with cell bursting and release of proinflammatory cytokines. A deeper understanding of AIH pathogenesis will contribute to novel therapy for AIH patients. Here, we aimed to investigate the role of IL-17 in immune-mediated liver injury.The levels of cytokines were measured by ELISA, and mRNA levels of STAT3 and IFN gammainducible protein 16 (IFI16) were detected by PCR. Expressions of STAT3, IFI16, gasdermin D and cleaved caspase-1 were measured by western-blotting. Immunohistochemical staining and transmission electron microscopy were applied to evaluate liver histopathological changes of the treated mice. Our results showed that the levels of IFI16 was increased in hepatocytes treated with IL-17 protein, and further elevated after STAT3-overexpressed (STAT3-OE) lentivirus treatment. The levels of IFI16 were reduced in hepatocytes treated with IL-17 neutralizing Ab (nAb), but were significantly increased after STAT3-OE treatment. Pyroptosis was observed in hepatocytes treated with IL-17 protein, and further cell damage was observed after STAT3-OE lentivirus treatment. Liver damage was alleviated in mice treated with IL-17 nAb, however sever damage was experienced after STAT3-OE lentivirus treatment. A binding interaction between IFI16 and STAT3 was detected in IL-17 treated hepatocytes. Glutathione transaminase activity was enhanced in concanavalin A-induced AIH mice compared to the control group (p<0.01). IL-17 plays an important role in activating STAT3 and up-regulating IFI16, which may promote the pyroptosis in AIH-related liver injury through STAT3-IFI16 axis.
4.Advances of the mechanism study on Chinese herb monomers in improving premature ovarian failure
Rugen YAN ; Yuqi WANG ; Jing HE ; Yin CHEN ; Pingping CAI ; Changzhong LI
China Pharmacy 2022;33(21):2685-2688
Premature ovarian failure (POF) is a common gynecological endocrine disease ,which can cause follicular development stagnation or premature failure in the ovary ,leading to ovarian function loss ,and seriously affecting women ’s physical and mental health . Traditional Chinese medicine shows less toxicity and side effects but significant effect in the treatment of POF . Chinese herb monomers are the active ingredients of traditional Chinese medicine that exert their effects ,and their chemical structure are determined . From the perspective of Chinese herb monomers ,the author reviewed the mechanism of their effect on improving POF . The results showed that icariin ,quercetin,resveratrol and ginsenoside Rg 1 could improve the ovarian function of POF model animals by activating the phosphatidylinositol 3-kinase/protein kinase B signaling pathway ,inhibiting oxidative stress and apoptosis of granulosa cells ,promoting ovarian autophagy ,and regulating the level of sex hormones ;puerarin and resveratrol can improve the ovarian function of POF model by interfering with Wnt/ β-catenin signaling pathway ,reducing oxidative stress response ,and inhibiting follicular atresia . Although the Chinese herb monomers have certain effect in improving POF,most of them remain in the experimental stage ,and the clinical efficacy in improving POF cannot be objectively evaluated . In the future ,the development of new dosage forms of Chinese herb monomers should be strengthened to lay a foundation for the development of new drugs for treating POF .
5.The effect of different HAART regimens on liver function and HIV load of HIV infected children with HCV co-infection
Heping RAO ; Changzhong JIN ; Xiangning JIN ; Meixin FANG ; Wei WANG ; Weili LU
Chongqing Medicine 2017;46(15):2045-2047
Objective To investigate the effect of different HAART regimens on liver function,viral load and CD4+T cells of HIV infected children with HCV co-infection.Methods A total of 40 patients were divided into 4 groups:13 cases in AZT+3TC+NVP group(group A),16 cases in AZT+3TC+EFV group(group B),5 cases in D4T+3TC+EFV group(group C)and 6 cases in D4T+3TC+NVP group(group D).Liver function,viral load and CD4+T cells of patients in 4 groups were compared between before and after HAART.Results Eight patients(20%)had abnormal liver function before HAART treatment,and all of them had 1 level of liver damage.After receiving HAART,levels of ALT and AST were increased in group A and B(P<0.01);AST levels were increased in group C(P<0.01).CD4+T counts were all increased and HIV viral loads were decreased in 4 groups(P<0.01).There was no significant difference in RNA HCV load before and after HAART treatment(P>0.05).Conclusion Different HAART regimens can increase the liver damage in HIV infected children co-infected with HCV,but have little effect on the treatment efficiency of HAART regimens.
6.IFN-αcould induce the expression of SAMHD1 by down-regulating miR-181a
Heping RAO ; Wei WANG ; Xiangning JIN ; Changzhong JIN
Chinese Journal of Microbiology and Immunology 2016;36(4):277-280
Objective To investigate whether the expression of sterile alpha motif and histidine/aspartic acid domain containing protein 1 ( SAMHD1 ) could be induced by IFN-α and mediated by microRNA-181a (miR-181a). Methods THP-1 and Jurkat cells were treated with different doses of IFN-α(200 IU/ml and 1 000 IU/ml) for 24 h. The expression of miR-181a and SAMHD1 at mRNA level were de-tected by quantitative PCR. Western blot assay was performed to measure the expression of SAMHD1 at pro-tein level. THP-1 and Jurkat cells were transfected with p-181a, an over-expression vector of miR-181a, and then treated with 200 IU/ml of IFN-α. Changes in the expression of SAMHD1 in those cells were analyzed. Results IFN-α significantly enhanced the expression of SAMHD1 at mRNA and protein levels, but inhibi-ted the expression of miR-181a, especially in Jurkat cells. The expression of SAMHD induced by IFN-αwas inhibited in cells over-expressing miR-181a. Conclusion This study suggests that IFN-α could induce the expression of SAMHD1 by down-regulating the level of miR-181a.
7.Influence of fenofibrate on tissue NF-KB, IL-6 and cell apoptosis in secondary brain injury after traumatic brain injury in rats
Changzhong SUN ; Yongqing WANG ; Yujiang PENG ; Bo SHAO ; Zhi YU
Journal of Chinese Physician 2015;(3):372-375
Objective To investigate the changes of NF-κB p65, IL-6 and cell apoptosis in sec-ondary brain injury after traumatic brain injury and the influence of fenofibrate on these parameters in rats. Methods Ninety-eight male Sprague-Dawley ( SD) rats were randomly divided into two groups:fenofibrate group ( n =49) and control group ( n =49) .The fenofibrate group was induced with the improved Feeney method and received intragastrica of lipanthyl 60 mg/(kg? d) immediately after injury.The control group were received intragastrica of sodium chloride injection 2 ml/( kg? d) immediately after injury and twice everyday until rats were killed.Each group was divided into seven subgroups by sacrificed time after injury, those were 1 h, 3 h, 6 h, 12 h, 24 h, 3 d, and 7 d, and each subgroup got 7 rats.Each subgroup was ran-domly selected three rats after being killed to detect expressions of NF-κB p65 and IL-6 of rat contusion peri tissues brain tissues with immunohistochemical method.While using terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling ( TUNEL) method was used to observe the peri cell apoptosis after brain contusion.Results The expressions of NF-κB p65 and the IL-6 in each fenofibrate group were significantly decreased relative to the control group ( P <0.05),and a significant positive correlation between both pa-rameters in two groups ( P <0.01) .At the same time, the number of apoptotic cells was decreased ( P <0.05).Conclusions Fenofibrate was probably through the route of relieving inflammation response to re-duce the change of secondary brain injury after traumatic brain injury and decrease neural cell apoptosis, and then provide protection of neurocytes.
8.Development of pyrogenation treatment vehicle of medical wastes
Lihua WU ; Zheng WANG ; Junshu HAN ; Wenchang ZHANG ; Chao WU ; Xiaojun SUN ; Xiangdong CUI ; Changzhong XU
Chinese Medical Equipment Journal 2015;36(5):98-101,106
Objective To develop a medical wastes pyrolysis vehicle for field hospital, countryside hospital and public emergencies.Methods Configuration design, thermodynamics theory, ergonomics and auto control technology were involved in to manufacture open-type compartment, on-board pyrolysis furnace, smoke control system, and pyrolysis furnace control and monitoring system.Results The vehicle passed 7 000 km reliability test, high-low temperature test, smoke emission test and army trials.Conclusion The vehicle behaves well in performances and environment protection, and gains high economic and social benefits.
9.Way of 'analytical thinking' on data from epidemiological studies.
Chinese Journal of Epidemiology 2014;35(6):745-748
Analysis on data from epidemiological studies is the sequencing process of applying statistical methods to collected data from different angles, interpreting intermediate results, drawing statistical conclusions and forming scientific findings based on existing knowledge. This is also called the 'process of converting data to evidence'. Final results from the analysis are expressed through scientific papers. Process of an accurate, clear and comprehensive data analysis is critical to form a convincing conclusion on a paper. This article discusses how to form the analytical thoughts for conducting a thorough data analysis in order to draw a convincing evidence from epidemiological data.
Data Interpretation, Statistical
;
Epidemiologic Studies
10.Way of‘analytical thinking’ on data from epidemiological studies
Chinese Journal of Epidemiology 2014;(6):745-748
Analysis on data from epidemiological studies is the sequencing process of applying statistical methods to collected data from different angles,interpreting intermediate results, drawing statistical conclusions and forming scientific findings based on existing knowledge. This is also called the‘process of converting data to evidence’. Final results from the analysis are expressed through scientific papers. Process of an accurate,clear and comprehensive data analysis is critical to form a convincing conclusion on a paper. This article discusses how to form the analytical thoughts for conducting a thorough data analysis in order to draw a convincing evidence from epidemiological data.

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