1.Chinese expert consensus on blood support mode and blood transfusion strategies for emergency treatment of severe trauma patients (version 2024)
Yao LU ; Yang LI ; Leiying ZHANG ; Hao TANG ; Huidan JING ; Yaoli WANG ; Xiangzhi JIA ; Li BA ; Maohong BIAN ; Dan CAI ; Hui CAI ; Xiaohong CAI ; Zhanshan ZHA ; Bingyu CHEN ; Daqing CHEN ; Feng CHEN ; Guoan CHEN ; Haiming CHEN ; Jing CHEN ; Min CHEN ; Qing CHEN ; Shu CHEN ; Xi CHEN ; Jinfeng CHENG ; Xiaoling CHU ; Hongwang CUI ; Xin CUI ; Zhen DA ; Ying DAI ; Surong DENG ; Weiqun DONG ; Weimin FAN ; Ke FENG ; Danhui FU ; Yongshui FU ; Qi FU ; Xuemei FU ; Jia GAN ; Xinyu GAN ; Wei GAO ; Huaizheng GONG ; Rong GUI ; Geng GUO ; Ning HAN ; Yiwen HAO ; Wubing HE ; Qiang HONG ; Ruiqin HOU ; Wei HOU ; Jie HU ; Peiyang HU ; Xi HU ; Xiaoyu HU ; Guangbin HUANG ; Jie HUANG ; Xiangyan HUANG ; Yuanshuai HUANG ; Shouyong HUN ; Xuebing JIANG ; Ping JIN ; Dong LAI ; Aiping LE ; Hongmei LI ; Bijuan LI ; Cuiying LI ; Daihong LI ; Haihong LI ; He LI ; Hui LI ; Jianping LI ; Ning LI ; Xiying LI ; Xiangmin LI ; Xiaofei LI ; Xiaojuan LI ; Zhiqiang LI ; Zhongjun LI ; Zunyan LI ; Huaqin LIANG ; Xiaohua LIANG ; Dongfa LIAO ; Qun LIAO ; Yan LIAO ; Jiajin LIN ; Chunxia LIU ; Fenghua LIU ; Peixian LIU ; Tiemei LIU ; Xiaoxin LIU ; Zhiwei LIU ; Zhongdi LIU ; Hua LU ; Jianfeng LUAN ; Jianjun LUO ; Qun LUO ; Dingfeng LYU ; Qi LYU ; Xianping LYU ; Aijun MA ; Liqiang MA ; Shuxuan MA ; Xainjun MA ; Xiaogang MA ; Xiaoli MA ; Guoqing MAO ; Shijie MU ; Shaolin NIE ; Shujuan OUYANG ; Xilin OUYANG ; Chunqiu PAN ; Jian PAN ; Xiaohua PAN ; Lei PENG ; Tao PENG ; Baohua QIAN ; Shu QIAO ; Li QIN ; Ying REN ; Zhaoqi REN ; Ruiming RONG ; Changshan SU ; Mingwei SUN ; Wenwu SUN ; Zhenwei SUN ; Haiping TANG ; Xiaofeng TANG ; Changjiu TANG ; Cuihua TAO ; Zhibin TIAN ; Juan WANG ; Baoyan WANG ; Chunyan WANG ; Gefei WANG ; Haiyan WANG ; Hongjie WANG ; Peng WANG ; Pengli WANG ; Qiushi WANG ; Xiaoning WANG ; Xinhua WANG ; Xuefeng WANG ; Yong WANG ; Yongjun WANG ; Yuanjie WANG ; Zhihua WANG ; Shaojun WEI ; Yaming WEI ; Jianbo WEN ; Jun WEN ; Jiang WU ; Jufeng WU ; Aijun XIA ; Fei XIA ; Rong XIA ; Jue XIE ; Yanchao XING ; Yan XIONG ; Feng XU ; Yongzhu XU ; Yongan XU ; Yonghe YAN ; Beizhan YAN ; Jiang YANG ; Jiangcun YANG ; Jun YANG ; Xinwen YANG ; Yongyi YANG ; Chunyan YAO ; Mingliang YE ; Changlin YIN ; Ming YIN ; Wen YIN ; Lianling YU ; Shuhong YU ; Zebo YU ; Yigang YU ; Anyong YU ; Hong YUAN ; Yi YUAN ; Chan ZHANG ; Jinjun ZHANG ; Jun ZHANG ; Kai ZHANG ; Leibing ZHANG ; Quan ZHANG ; Rongjiang ZHANG ; Sanming ZHANG ; Shengji ZHANG ; Shuo ZHANG ; Wei ZHANG ; Weidong ZHANG ; Xi ZHANG ; Xingwen ZHANG ; Guixi ZHANG ; Xiaojun ZHANG ; Guoqing ZHAO ; Jianpeng ZHAO ; Shuming ZHAO ; Beibei ZHENG ; Shangen ZHENG ; Huayou ZHOU ; Jicheng ZHOU ; Lihong ZHOU ; Mou ZHOU ; Xiaoyu ZHOU ; Xuelian ZHOU ; Yuan ZHOU ; Zheng ZHOU ; Zuhuang ZHOU ; Haiyan ZHU ; Peiyuan ZHU ; Changju ZHU ; Lili ZHU ; Zhengguo WANG ; Jianxin JIANG ; Deqing WANG ; Jiongcai LAN ; Quanli WANG ; Yang YU ; Lianyang ZHANG ; Aiqing WEN
Chinese Journal of Trauma 2024;40(10):865-881
Patients with severe trauma require an extremely timely treatment and transfusion plays an irreplaceable role in the emergency treatment of such patients. An increasing number of evidence-based medicinal evidences and clinical practices suggest that patients with severe traumatic bleeding benefit from early transfusion of low-titer group O whole blood or hemostatic resuscitation with red blood cells, plasma and platelet of a balanced ratio. However, the current domestic mode of blood supply cannot fully meet the requirements of timely and effective blood transfusion for emergency treatment of patients with severe trauma in clinical practice. In order to solve the key problems in blood supply and blood transfusion strategies for emergency treatment of severe trauma, Branch of Clinical Transfusion Medicine of Chinese Medical Association, Group for Trauma Emergency Care and Multiple Injuries of Trauma Branch of Chinese Medical Association, Young Scholar Group of Disaster Medicine Branch of Chinese Medical Association organized domestic experts of blood transfusion medicine and trauma treatment to jointly formulate Chinese expert consensus on blood support mode and blood transfusion strategies for emergency treatment of severe trauma patients ( version 2024). Based on the evidence-based medical evidence and Delphi method of expert consultation and voting, 10 recommendations were put forward from two aspects of blood support mode and transfusion strategies, aiming to provide a reference for transfusion resuscitation in the emergency treatment of severe trauma and further improve the success rate of treatment of patients with severe trauma.
2.Bonding strength of metal brackets to polished and glazed zirconia ceramics with 3 adhesives respectively
Yu CAO ; Jia LIU ; Huichao ZHANG ; Jingyu XU ; Ling YU ; Surong YE ; Luyi YANG
Journal of Practical Stomatology 2024;40(1):53-57
Objective:To investigate the effects of 3 adhesives on the bond force and durability of polished and glazed zirconia ceram-ics to orthodontic metal brackets respectively.Methods:Universal adhesives,Single Bond Universal(SBU)and Prime&Bond Universal(PBU)were respectively used to bond polished and glazed zirconia to metal braces of maxillary central incisors using TransbondTM MIP(TM)as the control.The shear bond strength(SBS),the fracture morphology and adhesive residual index(ARI)were examed after wa-ter bath or water bath-thermal cycling storage.Results:The adhesive(P<0.001)and storage conditions(P<0.001)significantly af-fected the shear bond strength of zirconia to brackets.There was no significant difference between the polished or glazed groups(P=0.09).SBU showed the stronger SBS and lower ABI,there were significant differences in ARI scores among the 3 cements(P<0.001).Conclusion:SBU may have better bonding performance than PBU and TM in the orthodontic bonding of polished or glazed zir-conia surfaces to the zirconia ceramics.
3.Application of near-infrared autofluorescence probe in intraoperative parathyroid gland identification
Surong HUA ; Junyi GAO ; Zhen CAO ; Huaijin ZHENG ; Hongyu WANG ; Xiaojing NING ; Liyuan FU ; Yang ZHANG ; Yikun WANG ; Ziwen LIU ; Quan LIAO
Chinese Journal of Endocrine Surgery 2024;18(5):675-678
Objective:To explore the use of near-infrared autofluorescence probe (NIRAF-P) and its application in identifying parathyroid glands during surgery.Methods:A total of 68 patients undergoing thyroid surgery at Peking Union Medical College Hospital and Beijing Longfu Hospital between Dec. 2023 and Jun. 2024 were selected. During the operation, the near-infrared parathyroid gland detector was used to identify the parathyroid gland tissue to be tested, and histopathological examination was performed. The positive predictive value and accuracy of the near-infrared parathyroid gland detector were analyzed.Results:A total of 111 parathyroid glands were identified in 68 patients, and the positive predictive value and accuracy of the NIRAF-P were 95.5% and 94.6%, respectively.Conclusions:The NIRAF-P has high accuracy in identifying parathyroid glands. The standardized application of the NIRAF-P can help improve the efficiency of identifying parathyroid glands during surgery.
4.Expression of lncRNA CCAT2 and OTUD1 in multiple myeloma tissue and their relationship with clinicopathological features and prognosis
Yang QUAN ; Peidong HE ; Jiao ZHU ; Hongcang WANG ; Surong LIU
Chongqing Medicine 2024;53(21):3215-3221
Objective To investigate the expression of long non-coding RNA(lncRNA)CCAT2 and o-varian cancer domain-containing protease 1(OTUD1)in multiple myeloma(MM)tissue and their correlation with clinicopathological characteristics and prognosis of MM.Methods A total of 132 patients with MM(MM group)diagnosed and treated in this hospital from April 2018 to April 2020 were selected.Seventy patients with non-hematological disease who underwent bone marrow puncture without abnormal bone marrow func-tion during the same period served as the control group.The real-time fluorescence quantitative PCR was used to detect the expression levels of lncRNA CCAT2 and OTUD1 mRNA in bone marrow tissue.The Pearson correlation was performed to analyze the correlation between lncRNA CCAT2 and OTUD1 mRNA expression in bone marrow tissues.The expression differences of lncRNA CCAT2 and OTUD1 mRNA were compared a-mong the MM patients with different clinical and pathological characteristics.The Kaplan-Meier curve was used to analyze the difference of prognosis among the MM patients with different lncRNA CCAT2 and OTUD1 mRNA expressions.The Cox regression was performed to analyze the factors affecting the prognosis in MM patients.Results The expression level of lncRNA CCAT2 in the bone marrow tissue of the MM group was significantly higher than that of the control group(2.31±0.67 vs.0.85±0.24),while the expression level of OTUD1 mRNA in the MM group was lower than that of the control group(1.22±0.37 vs.2.54±0.75),and the differences were statistically significant(t=17.624,16.760,all P<0.001).The lncRNA CCAT2 expression level in the bone marrow tissue of the MM group had significantly negative correlation with the OTUD1 mRNA expression level(r=-0.731,P<0.001).The lncRNA CCAT2 and OTUD1 mRNA expression levels had statistical differences among the MM patients with different ISS stages and β2-micro-globulin levels(P<0.001).The 3-year overall survival rates of the high and low expression groups of ln-cRNA CCAT2 were 42.19%(27/64)and 66.18%(45/68),respectively.The 3-year overall survival rates of the high and low expression groups of OTUD1 mRNA were 72.31%(47/65)and 37.31%(25/67)respective-ly.The 3-year cumulative survival rate of MM patients in the lncRNA CCAT2 low expression group was sig-nificantly higher than that in the lncRNA CCAT2 high expression group,and the difference was statistically significant(Log Rank X2=7.151,P=0.007).The 3-year cumulative survival rate of MM patients in the OTUD1 mRNA low expression group was significantly lower than that in the OTUD1 mRNAhigh expression group(Log Rank x2=13.667,P<0.001).The ISS stage Ⅲ and lncRNA CCAT2 high expression were the risk factors affecting the prognosis of MM patients(P<0.01),while the OTUD1 mRNA high expression was the protective factor.Conclusion The lncRNA CCAT2 expression level in bone marrow tissue of the MM pa-tients is increased and OTUD1 expression level is decreased,the both are associated with adverse clinical and pathological characteristics of MM and independent factors affecting the prognosis of MM patients.
5.Genomic epitopes of Xinjiang hemorrhagic fever virus and the evaluation of its laboratory testing value
Huahua ZHANG ; Guoyu ZHAO ; Yang LI ; Shen SHI ; Ran AN ; Jiangling YUAN ; Dongliang LIU ; Jingrui DOU ; Tao LUO ; Surong SUN ; Yujiang ZHANG
Chinese Journal of Laboratory Medicine 2023;46(2):127-136
Objective:To grasp the distribution of fine antigenic epitope profiles of nucleoprotein (NP) and glycoprotein (GP) fragments of Crimean-Congo hemorrhagic fever virus (CCHFV) and to clarify the value of dominant antigenic epitopes in laboratory testing of Crimean-Congo hemorrhagic fever (CCHF).Methods:In a minimal synthetic short peptide consisting of 8 amino acids was segmentally expressed by CCHFV YL04057 strain using a modified bio-peptide synthesis method from 2014 to 2021 in the laboratory of Xinjiang University, College of Life Sciences. Using CCHFV polyclonal antibody or monoclonal antibody 14B7 (IgM) or CCHFV-positive sheep serum as antibodies, the minimal antigenic epitopes (BCEs) with antigenic activity on NP and GP fragments were identified by immunoblotting, and the obtained BCEs with sequence polymorphism were spatially clustered with CCHFV from different regions using the neighbor-joining method to determine the combination mode of BCEs with geographical correlation of regional distribution, to explore its application in establishing serological diagnosis. A prokaryotic expression plasmid (pET-32a), an E. coli expression plasmid (pGEX-KG) and a prokaryotic expression plasmid with an incomplete glutathione (GST188) tag (pXXGST-ST-1) were used to construct and express six dominant antigenic epitopes of different peptide lengths on NP fragments, and an indirect Enzyme-linked immunosorbent assay (ELISA) was established. CCHF sheep serum identified by immunofluorescence assay (IFA) was used as a control, and the specificity, sensitivity and overall compliance of the recombinant proteins with different peptide lengths of antigenic epitopes with IFA assay results were statistically analyzed. Results:CCHFV, NP and GP fragments had a total of 30 antigenically active BCEs, among which the core intermediate fragment NP2 (aa 170 th-305 th), which had a concentration of antigenic epitopes in the NP fragment, has 6 BCEs, and the NP1 (aa 1 st-200 th) and NP3 (aa 286 th-482 nd) at both ends have 9 BCEs; the Gc (aa 1 st-558 th) and Gn (aa 533 th-708 th) fragments of the GP fragment have 14 BCEs and a long antigenic peptide (AP) containing 15 amino acids, and the amino acid sequence homology of the NP fragment BCEs was 97.1% and that of the GP fragment BCEs was 89.1%. There was a significant difference ( P=0.0281, P<0.05). Among the 9 BCEs with sequence polymorphism in the GP fragment, 6 combined BCEs from GnEc1, GnE2, GnE4, GcE3, GcE6 and GcAP-4 (Ap) could cluster 15 CCHFV strains from different regions of the world into 5 geographical taxa, AsiaⅠ, AsiaⅡ, AficaⅠ, AficaⅡ and Europe. The constructs expressing PET-32a-NP (full length), PGEX-KG-NP2 (aa 170 th-305 th), pGEX-KG-NP2-1 (aa 235 th-275 th), PGEX-KG-NP2-1-1 (aa 237 th-256 th), pXXGST-1-NP2-1-2 (aa 250 th-265 th) and PGEX KG-NP2-1-3 (aa 260 th-276 th), six recombinant proteins CCHFV NP rabbit polyclonal antiserum (pAb) Western Blotting reaction positive, 33 sheep sera tested by IFA XHF as a reference, the sensitivity of the assay established by indirect ELISA using the recombinant proteins constructed from two fragments of NP2 and NP2-1 as antigens. The sensitivity, specificity and overall compliance were the best, with 73.4% (11/15) and 66.7% (10/15) for sensitivity, 100% (18/18) and 94.4% (17/18) for specificity, and 87.9% (29/33) and 81.8% (27/33) for overall compliance. Conclusion:CCHFV NP and GP are distributed with a high number of BCEs with antigenic immunoreactivity, among which the dominant antigenic epitopes are of high value in the laboratory serological diagnosis of CCHF.
6.Application of Endoscopic Parathyroidectomy in the Treatment of Primary Hyperparathyroidism
Surong HUA ; Zhihong WANG ; Junyi GAO ; Mengyi WANG ; Qiaofei LIU ; Wenjing LIU ; Guannan GE ; Yingxin WEI ; Ya HU ; Quan LIAO
Chinese Journal of Endocrine Surgery 2022;16(4):391-395
Objective:To summarize the experience and the clinical data of patients with primary hyperparathyroidism undergoing endoscopic parathyroidectomy.Methods:A total of 24 patients who underwent endoscopic parathyroidectomy for primary hyperparathyroidism in Peking Union Medical College Hospital during Feb. 2021 to May. 2022 were concluded in this study (20 cases of parathyroidectomy via axillary approach and 4 cases of parathyroidectomy via thoracic and breast approach) . The operation time, postoperative drainage, length of stay, level of parathyroid hormone and serum calcium of those patients were collected. Postoperative complications and recurrence of hyperparathyroidism were also observed.Results:The postoperative levels of serum parathyroid hormone and serum calcium were significantly reduced (over 50%) compared with preoperative level ( P<0.05) . The average operation time was (96±22) min (64-157 min) . The mean postoperative drainage volume was (47±16) ml on day 1, (46±11) ml on day 2, and (30±9) ml on day 3, respectively. The average length of postoperative hospital stay was (2.8±1.1) days (2-6 days) . In one case of parathyroidectomy via axillary approach, the operation was converted to open surgery because of the low position of lesion. Other cases completed endoscopic surgery and obtained satisfactory cosmetic results. There were no postoperative complications such as bleeding, permanent hoarseness, coughing while drinking water, or surgical site infection. The mean follow-up time was (7.4±4.2) months (1-16 months) . There was no obvious discomfort and no recurrence during follow-up. Conclusion:Endoscopic parathyroidectomy is safe and effective in the treatment of primary hyperparathyroidism, which can be used as a surgical option for patients with cosmetic requirements.
7.Analysis of influencing factors for the development of gallstone in population of Beijing
Zhihong WANG ; Jiayi LI ; Surong HUA ; Bo TANG ; Tao HONG ; Wei LIU ; Xiaodong HE
Chinese Journal of Digestive Surgery 2022;21(7):910-916
Objective:To investigate the influencing factors for the development of gall-stone in population of Beijing.Methods:The retrospective cross-sectional survey was conducted. From November 2016 to September 2020, patients living in Beijing (registered residence in Beijing ≥12 months) who visited the biliary outpatient of Department of General Surgery of Peking Union Medical College Hospital, Chinese Academy of Medical Sciences for the first time were recruited to participate as respondents. The survey was conducted by the questionnaire survey on correlation between dietary habits and incidence of gallstones, in which the information of gender, age, body mass index (BMI), gallstone status, metabolic indicators (hypercholesterolemia, history of diabetes mellitus, reproductive times for female, menopause status of female, duration of menopause for female, history of weight loss), dietary indicators (dietary mix of meat and vegetable dishes, times of coffee intake per month, times of alcohol consumption per month, times of greasy diet intake per month, times of breakfast skipping per week, average overnight fasting time of breakfast skipping, times of supper skipping per week, average overnight fasting time of supper skipping), family history of gallstones, lifestyle indicators (times of staying up late per month, average overnight fasting time when staying up late, daily sedentary time, weekly physical activity score). Observation indicators: (1) results of questionnaire survey; (2) analysis of influencing factors for the occurrence of gallstone. Measurement data with normal distribution were represented as Mean± SD, and measurement data with skewed distribution were represented as M( Q1, Q3). Count data were expressed as absolute numbers or percentages. Univariate and multivariate analyses were performed using the Logistic regression model. Results:(1) Results of questionnaire survey. A total of 1 036 questionnaires were distributed, and 1 004 complete questionnaires were recovered. Of the 1 004 patients who completed the questionnaire survey, there were 329 males and 675 females, aged (44±12)years. The BMI of 1 004 patients was (24±3)kg/m 2. Of the 1 004 patients, there were 659 cases with a history of cholecystolithiasis and 345 cases without. (2) Analysis of influencing factors for the occurrence of gallstone. Results of univariate analysis showed that age, history of diabetes mellitus, history of weight loss, times of coffee intake per month, times of greasy diet intake per month, family history of gallstone and daily sedentary time were related factors for the development of gallstone in 1 004 patients ( odds ratio=1.03, 2.26, 1.74, 1.01, 1.01, 2.22, 1.06, 95% confidence intervals as 1.02?1.05, 1.09?5.18, 1.22?2.53, 1.00?1.03, 1.00?1.01, 1.60?3.11, 1.01?1.11, P<0.05). Results of multivariate analysis showed that age, history of diabetes mellitus, history of weight loss, times of greasy diet intake per month, family history of gallstone and daily sedentary time were independent influencing factors for the development of gallstone in 1 004 patients ( odds ratio=1.03, 2.26, 1.82, 1.01, 2.22, 1.06, 95% confidence intervals as 1.02?1.05, 1.11?5.13, 1.28?2.62, 1.00?1.02,1.60?3.09, 1.01?1.12, P<0.05). Conclusion:Age, history of diabetes mellitus, history of weight loss, times of greasy diet intake per month, family history of gallstone and daily sedentary time are independent influencing factors for the development of gallstone in population of Beijing.
8.Application of precision nursing emergency management in emergency rescue of emergencies
Shanshan YANG ; Surong LIU ; Li XU ; Hong XIAO ; Yan YE ; Li HUANG ; Shuhua DENG
Chinese Journal of Practical Nursing 2020;36(34):2695-2700
Objective:To explore the management effectiveness of emergency treatment management for emergencies.Methods:To establish precision nursing emergency management system, the response time, triage accuracy, receiving time, information delivery time and rescue success rate were compared before and after the application of the system.Results:After the application of the precision nursing emergency management, the emergency response time was shortened from (6.47±1.25) min to (3.56±1.38) min, and the time for admission and triage reduced from (5.15±0.54) min to (2.84±0.49) min. The time was shortened from (92.45±10.49) minutes to (72.35±13.20) minutes, and the time for information submission was shortened from (121.47±58.41) minutes to (65.23±10.72) minutes; the accuracy of triage diagnosis increased from 96.85% (277/286) to 99.27%(271/273). The rescue success rate increased from 96.15%(275/286) to 98.90%(270/273), and the differences were statistically significant ( t value was -2.920-5.587, χ2 value was 4.220, 4.317, P<0.05). Conclusions:The application of precision nursing emergency management in emergencies can improve the emergency response rate and overall level of nursing staff and ensure the safety of patients.
9.Expression and antibody preparation of recombinant truncated glycoprotein of Guertu virus
Abula AYIPAIRI ; Shu SHEN ; Jingyuan ZHANG ; Xijia LIU ; Yijie LI ; Fei DENG ; Yujiang ZHANG ; Surong SUN
Chinese Journal of Microbiology and Immunology 2020;40(3):178-184
Objective:To express truncated glycoprotein (Gn, Gn1, Gn2, Gn3, Gc1 and Gc2) of Guertu virus (GTV) in Escherichia coli ( E. coli) cells, and prepare polyclonal antibodies against recombinant proteins Gn-His, Gc1-His and Gc2-His after purification. Methods:Gene fragments encoding Gn, Gn1, Gn2, Gn3, Gc1 and Gc2 of GTV DXM strain were amplified by RT-PCR, and cloned into the prokaryotic expression vector pET-32a (+ ) to construct recombinant expression plasmids. The transformed E. coli BL21(DE3) strains carrying expression plasmids were induced by IPTG to express target proteins, which were identified by SDS-PAGE. Recombinant proteins Gn-His, Gc1-His and Gc2-His were purified by nickel affinity chromatography and detected by Western blot using GTV-positive sheep serum for analysis of their antigenicity. New Zealand white rabbits were immunized with the purified recombinant proteins. The titers and specificity of serum antibodies were analyzed by ELISA. Meanwhile, eukaryotic expression vectors pcDNA3.1-Gn, pcDNA3.1-Gc1/Gc2 were constructed and transfected into mammalian Vero cells to evaluate the binding activity of rabbit polyclonal antibodies by indirect immunofluorescence method. The specific reactivity of serum antibodies to recombinant proteins was detected by Western blot. Results:Restriction enzyme analysis and DNA sequencing confirmed that the recombinant expression vectors of pET-32a-Gn, pET-32a-Gn1/Gn2/Gn3, pET-32a-Gc1/Gc2, pcDNA3.1-Gn and pcDNA3.1-Gc1/Gc2 were constructed successfully. The relative molecular mass ( Mr) of the expressed recombinant proteins Gn-His, Gn1/Gn2/Gn3-His, Gc1/Gc2-His were approximately 63.4×10 3, 37.1×10 3, 31.9×10 3, 30.8×10 3, 40×10 3 and 54.4×10 3, respectively. The recombinant proteins could be recognized by GTV-positive sheep serum. The titers of polyclonal antibodies against GTV Gn, Gc1 and Gc2 were 1∶409 600, 1∶204 800 and 1∶6 400, respectively. Indirect immunofluorescence assay and Western blot showed that the prepared rabbit polyclonal antibodies could specifically react with the proteins expressed in eukaryotic cells and the recombinant proteins. Conclusions:The recombinant GTV glycoproteins Gn-His and Gc1/Gc2-His were efficiently expressed and purified and characterized with good immunity. The prepared polyclonal antibodies had high titers and good specificity. This study provided reference for further studying the biological function and detection methods of GTV glycoproteins and research on vaccines.
10.Down-regulation of miR-205-5p enhances pro-apoptotic effect of 3-bromopyruvate on human nasopharyngeal carcinoma CNE2Z cells.
Zongfen SHI ; Pei ZHANG ; Xingyue LU ; Chenlu ZHU ; Changjiang CHEN ; Surong ZHAO ; Hao LIU
Journal of Southern Medical University 2019;39(10):1166-1172
OBJECTIVE:
To investigate the effect of down-regulation of miR-205-5p on 3-bromopyruvate-induced apoptosis in human nasopharyngeal carcinoma CNE2Z cells.
METHODS:
Nasopharyngeal carcinoma CNE2Z cells were transfected with miR- 205-5p-mimic or miR-205-5p-inhibitor, treated with 80 μmol/L 3-bromopyruvate alone, or exposed to both of the treatments. The proliferation of the treated cells was examined with MTT assay, and early apoptosis of the cells was detected using a mitochondrial membrane potential detection kit (JC-1). DAPI fluorescence staining was used to detect morphological changes of the cell nuclei and late cell apoptosis; Annexin V-FITC/PI double staining was employed to detect the cell apoptosis rate. Western blotting was used to detect the expressions of Bcl-2, Bax, Mcl-1 and Bak proteins.
RESULTS:
Exposure to 3-bromopyruvate significantly inhibited the proliferation of CNE2Z cells, and increasing the drug concentration and extending the treatment time produced stronger inhibitory effects. Treatment with 80 μmol/L 3-bromopyruvate for 24, 48 and 72 h resulted in inhibition rates of (45.7±1.21)%, (64.4±2.02)% and (78.3±1.55)% in non-transfected CNE2Z cells, respectively; the inhibition rates were (27.7±1.04)%, (34.8±2.10)% and (44.3±1.57)% in the cells transfected with miR-205-5p-mimic, and were (80.5 ± 0.94)%, (87.9 ± 0.50)% and (93.8 ± 1.16)% in cells transfected with miR-205-5p-inhibitor, respectively. The results of mitochondrial membrane potential detection showed that the relative proportion of red and green fluorescence decreased significantly in miR-205-5p-inhibitor-transfected cells with 3-bromopyruvate treatment. Combined treatment of the cells with 3-bromopyruvate and miR-205-5p-inhibitor transfection obviously increased nuclear fragmentation and nuclear pyknosis and significantly increased cell apoptotic rate as compared with the two treatments alone ( < 0.01), causing also decreased expressions of Bcl-2 and Mcl-1 proteins and increased expressions of Bax and Bak proteins.
CONCLUSIONS
Inhibition of miR-205-5p enhances the proapototic effect of 3-bromopyruvate in CNE2Z cells possibly in relation to the down-regulation of Mcl-1 and Bcl-2 and the up-regulation of Bak and Bax proteins.

Result Analysis
Print
Save
E-mail