1.Design,preparation,and antitumor activity of fusion protein vaccine based on tumor antigen PBK
Qianqian QIAN ; Guozhi LI ; Hong TIAN ; Xiangdong GAO
Journal of China Pharmaceutical University 2024;55(5):657-665
PBK is a cancer/testis antigen that exhibits absent expression in various normal human tissues,but undergoes aberrant overexpression upon cellular transformation,thereby promoting the initiation,metastasis and even drug resistance of cancer.Consequently,it represents a novel target for tumor immunotherapy.In this study,PBK-Nitrath,a protein vaccine specifically designed to target PBK by fusing nitrated T epitope with the PBK protein was developed,using the IFN-γ ELISpot method to evaluate the activation level of PBK antigen-specific T cells in the spleen of immunized mice,and conducting in vitro cytotoxicity T cell killing efficacy test to evaluate the killing ability against H22 hepatic carcinoma cells;the anti-tumor activity was evaluated using a H22 hepatic carcinoma subcutaneous transplantation tumor model,and the differentiation of peripheral blood and spleen T lymphocytes and tumor immune infiltration were analyzed by flow cytometry.Results showed that PBK-Nitrath could efficiently induce the activation of antigen-specific T cells against PBK while enhancing cytotoxic T lymphocyte-mediated killing capacity,significantly impede hepatic carcinoma progression in mice and increase the ratio of CD8+CD107a+T cells within peripheral blood and spleen,and facilitate tumor lymphocyte infiltration.Our findings reveal the potential utility of PBK-Nitrath as an effective candidate for tumor vaccine.
2.Cross-sectional survey on knowledge, attitude, behavior and infection of brucellosis in relocated poverty alleviation and relocation area in Tongyu County, Jilin Province
Biao LI ; Guozhi WEI ; Mujinyan LI ; Fande LI ; Taijun WANG ; Rui HUO ; Qing ZHEN
Chinese Journal of Endemiology 2022;41(10):831-836
Objective:To learn about the knowledge, attitude and behavior of brucellosis and brucellosis infection in Tongyu County, Jilin Province, and to provide a reference for the formulation of prevention and control measures in relocated poverty alleviation and relocation areas.Methods:Using a multi-stage sampling method, three townships, Xianghai Township, Wulanhua Town and Xinhua Town were selected in Tongyu County, where the incidence of brucellosis is high in Jilin Province; then Miren, Huimin, Longjing, Xinfeng and Dayou villages from the three townships were surveyed. A face-to-face questionnaire survey was conducted among the villagers to collect the knowledge, attitude and behavior of brucellosis; and blood samples were collected from the villagers according to the principle of informed consent for brucellosis serological tests, and according to the "Diagnostic criteria for Brucellosis (WS 269-2007)" for diagnosis.Results:A total of 274 questionnaires were distributed and 244 valid questionnaires were collected with a recovery rate of 89.05%. Among them, 233 people had heard of brucellosis, accounting for 95.49%. The total awareness rate of knowledge related to brucellosis prevention and control was 39.29% (2 205/5 612), of which the total awareness rate of "knowledge related to brucellosis prevention" was 71.99% (527/732), and the total awareness rate of "knowledge related to human infection with brucellosis" was 38.08% (1 115/2 928), and the total awareness rate of "knowledge about brucellosis in animals" was 28.84% (563/1 952). A total of 90.13% (21/233) people supported the brucellosis prevention and control plan (such as free screening for brucellosis, publicity and education, immunization, culling, etc.), and 61.54% (48/78) of the people would take the initiative to buy protective equipment. The contact rates of livestock through assisted feeding and slaughtering were 27.46% (67/244) and 11.48% (28/244), respectively, and the total protection rates of sheep pens cleaning and slaughtering were 30.91% (102/330) and 21.43% (30/140), respectively. In terms of brucellosis infection, a total of 1 confirmed case, 1 latent infection and 37 suspected cases were found.Conclusions:The awareness rate of "knowledge related to brucellosis prevention" in Tongyu County is generally good, but the awareness rate of "knowledge about brucellosis in animals" is low, and the protection rate of some brucellosis prevention and control behaviors is poor. Health and epidemic prevention departments should carry out targeted health education, improve and optimize propaganda methods in order to improve the protection level of the general population and reduce the risk of brucellosis infection.
3.H. L.P.E.S. scoring system combined with a virtual reality technique for evaluation of the stone-free rate of flexible ureteroscopy in the treatment of renal calculi
Jianglin GU ; Shengjun LUO ; Li JIANG ; Daixing HU ; Guozhi ZHAO ; Wei TANG
Chinese Journal of Urology 2022;43(5):384-386
The clinical data of 120 patients admitted to our hospital with renal stones treated by flexible ureteroscopy (FURS) and the imaging-related data measured by virtual reality technology were retrospectively analysed. The results of the univariate analysis showed that stone surface area (S), renal pelvis volume (P), length of calyces funnel (L), pelvic calyceal height (H) and essence of stone (E) were closely related to stone-free rate. The H. L.P.E.S. score was constructed to predict stone-free rate after FURS based on the above factors, and the area under the receiver aperating characteristic curve for the H. L.P.E.S. and S. O.L.V.E. scoring systems was 0.921 and 0.754 respectively.The H. L.P.E.S. scoring system has higher predictive value.
4.Ureaplasma urealyticum GrpE promotes mouse dendritic cell maturation and elicits Th1 immune responses
Fangyi GUO ; Yanhong TANG ; Hongxia YUAN ; Wenjun ZHANG ; Jing XIANG ; Pengqin LIU ; Wenyou TENG ; Ranhui LI ; Guozhi DAI
Chinese Journal of Microbiology and Immunology 2022;42(1):41-49
Objective:To investigate the effects of Ureaplasma urealyticum GrpE ( Uu-GrpE) on the maturation of dendritic cells and the polarization of T cells. Methods:Uu-GrpE was expressed and purified, and then identified by Western blot. The cytotoxicity of Uu-GrpE to mouse bone marrow-derived dendritic cells (BMDCs) was analyzed by LDH kit. After stimulating BMDCs with Uu-GrpE, the expression of costimulatory molecules, CD80, CD86 and major histocompatibility complex Ⅱ (MHCⅡ), on the surface of BMDCs was detected by flow cytometry, and ELISA was used to detect the cytokines such as IL-12p70, TNF-α, IL-1β and IL-6. CD4 + Na?ve T cells were isolated from mouse spleen tissues by magnetic beads. A co-culture system of BMDCs and Na?ve T cells was constructed to analyze the effects of GrpE-stimulated mature BMDCs (GrpE-BMDCs) on T cell proliferation and polarization towards Th1/Th2. Mice were immunized with GrpE-BMDCs through the tail vein, and the induced humoral and cellular immune responses were detected by ELISA and flow cytometry. Results:Uu-GrpE was successfully express and high purity BMDCs were isolated. Uu-GrpE could stimulate BMDCs to secrete cytokines such as IL-12p70, TNF-α, IL-1β and IL-6 without having cytotoxicity. Uu-GrpE significantly increased the expression of CD80 [mean flourscence indensity (MFI): (324.00±22.11) vs (91.03±10.95), P<0.01], CD86 [MFI: (1 176.00±51.39) vs (217.00±14.93), P<0.01] and MHCⅡ [MFI: (708.70±56.32) vs (185.70±16.77), P<0.01] on BMDCs. Compared to the GrpE-BMDCs only group and GrpE (boiled)-BMDCs+ T cell group, the GrpE-BMDCs+ T cell group showed significantly increased T cell proliferation [stimulation index: (7.25±0.21) vs(6.55±0.23) and (6.09±0.35), both P<0.05], and dramatically promoted T cell secretion of IL-2 and IFN-γ [IL-2: (145.60±14.67) pg/ml vs(55.92±3.12) pg/ml and (26.05±2.40) pg/ml, P<0.05 and P<0.01; IFN-γ: (267.20±37.80) pg/ml vs(146.70±20.65) pg/ml and(27.84±6.69) pg/ml, both P<0.05]. However, no significant change was observed in the expression of Th2-type cytokines. Moreover, the adoptive transfer of GrpE-BMDCs induced a Th1-type immune response. Conclusions:Uu-GrpE could stimulate the maturation and polarization of BMDCs. Moreover, it could induce Th1 immune response as a candidate protein vaccine for Ureaplasma urealyticum.
6.Comparison of therapeutic effect between Kirschner wire-loop plating and clavicular hook plate in the treatment of distal clavicle fracture
Bin ZHU ; Zhiming QI ; Yiteng WANG ; Menghong YIN ; Guozhi WU ; Hengshan LIU ; Qing LI
Chinese Journal of Postgraduates of Medicine 2017;40(3):214-218
Objective To compare the therapeutic effect between Kirschner wire- loop plating and clavicular hook plate in the treatment of distal clavicle fractures. Methods Forty- two patients with type Neer Ⅱ distal clavicle fracture were selected, among whom 23 patients were treated with Kirschner wire- loop plating (group A), and 19 patients were treated with clavicular hook plate (group B). The operative time, fluoroscopy time, length of incision, intra- operative bleeding volume and hospitalization time etc were recorded. The patients were followed up at 6 weeks, 3 months, 6 months and 12 months after operation. The American shoulder and elbow society (ASES) score was evaluated, and the therapeutic effect was evaluated by Karlsson standard. Results The operative time and fluoroscopy time in group B were significantly lower than those in group A: (63.5 ± 12.4) min vs. (71.5 ± 11.1) min and (3.6 ± 1.9) s vs. (4.8 ± 1.5) s, but the intra- operative bleeding volume, length of incision and cases of postoperative shoulder pain in group A were significantly lower than those in group B:(91.5 ± 18.5) ml vs. (108.7 ± 32.5) ml, (6.6 ± 0.7) cm vs. (11.3 ± 0.9) cm and 2 cases vs. 14 cases, and there were statistical differences (P<0.05 or <0.01). There was no statistical difference in hospitalization time between 2 groups (P>0.05). There was no statistical difference in ASES score at 6 weeks and 3 months after operation between 2 groups (P>0.05); but the ASES score at 6 and 12 months in group A was significantly higher than that in group B: (89.9 ± 12.6) scores vs. (79.7 ± 12.6) scores and (96.1 ± 4.8) scores vs. (86.1 ± 12.4) scores, and there was statistical difference (P<0.05 or <0.01). There was no statistical difference in the cases of good and excellent at 6 weeks, 3 months and 6 months after operation (P>0.05);but the cases of good and excellent at 12 months in group A was significantly more than that in group B:23 cases vs. 13 cases, and there was statistical difference (P<0.05). Conclusions Kirschner wire- loop plating provides a new treatment option for type Neer Ⅱ distal clavicle fractures. It is conducive to decrease the incidence of shoulder pain and improve the range of motion of should joint. Besides, this technology has advantage of small incision, relatively simple operation steps and small damage to the surrounding soft tissues of the acromioclavicular joint. It may be superior to the clavicular hook plate.
7.Study on the regulation mechanism of specific proteins c-kit of interstitial cells of Cajal in the pathogenesis of slow transit constipation in rats
Zhihao LIU ; Shuguang LI ; Xiangyang YU ; Jianli CHEN ; Wenbin CAO ; Yongkui ZHAO ; Guozhi ZHANG
Journal of Clinical Medicine in Practice 2017;21(23):43-45
Objective The explore regulation mechanism of specific proteins c-kit of interstitial cells of Cajal in the pathogenesis of slow transit constipation in rats.Methods Slow transit constipation in rat model was duplicated by injections of morphine (25 mg/kg,45 d).The number and weight of fecal granule were assayed.Daily average number of fecal particles,quality,and the time for first black discharge were analyzed.At the same time,the c-kit and SCF expression were detected by Real-time PCR and Western blot.The changes of inflammatory factors such as IL-1β,IL-2 and IL-10 were also detected by ELISA.Results Daily average number of fecal particles in constipation group was decreased,quality was higher,and the first time for first black discharge was prolonged,which showed significant differences (P < 0.05).The expression levels of c-kit and SCF were decreased significantly,and levels of inflammatory factors IL-1β,IL-2 and IL-10 were higher in constipation rats than the controls (P < 0.05).Conclusion The pathogenic progress of slow transit constipation rats are regulated,which is also related with inflammatory reactions.
8.Study on the regulation mechanism of specific proteins c-kit of interstitial cells of Cajal in the pathogenesis of slow transit constipation in rats
Zhihao LIU ; Shuguang LI ; Xiangyang YU ; Jianli CHEN ; Wenbin CAO ; Yongkui ZHAO ; Guozhi ZHANG
Journal of Clinical Medicine in Practice 2017;21(23):43-45
Objective The explore regulation mechanism of specific proteins c-kit of interstitial cells of Cajal in the pathogenesis of slow transit constipation in rats.Methods Slow transit constipation in rat model was duplicated by injections of morphine (25 mg/kg,45 d).The number and weight of fecal granule were assayed.Daily average number of fecal particles,quality,and the time for first black discharge were analyzed.At the same time,the c-kit and SCF expression were detected by Real-time PCR and Western blot.The changes of inflammatory factors such as IL-1β,IL-2 and IL-10 were also detected by ELISA.Results Daily average number of fecal particles in constipation group was decreased,quality was higher,and the first time for first black discharge was prolonged,which showed significant differences (P < 0.05).The expression levels of c-kit and SCF were decreased significantly,and levels of inflammatory factors IL-1β,IL-2 and IL-10 were higher in constipation rats than the controls (P < 0.05).Conclusion The pathogenic progress of slow transit constipation rats are regulated,which is also related with inflammatory reactions.
9.Evaluation of Fractional Flow Reserve Determined by CT Coronary Angiography in Relevant Patients
Di FAN ; Guangbin CUI ; Qiang LI ; Jia ZHU ; Wei WANG ; Guozhi XIA ; Yu XIANG
Chinese Circulation Journal 2016;31(9):840-843
Objective: To evaluate the accuracy and clinical value of fractional lfow reserve (FFR) determined by CT coronary angiography (CTA) in relevant patients. Methods: A total of 43 patients treated in our hospitals from 2013-10 to 2015-10 were retrospectively studied. There were 29 (67.40%) with male gender, the average age was (60.2±10.1) years. The patients received CTA at 1 week prior coronary angiography (CAG), the interval between CTA and CAG was (5.4±1.6) days. FFR was measured by both CAG and CTA (FFRCT) in selected target vessel which was deifned as maximal diameter reduction 50% to 70%. The imaging data were recorded and compared, FFRCT was calculated. Results: 48 vessels from 43 patients were eligible for analysis as target vessels. FFRCT vas evaluated based on the gold criteria of FFR. FFRCT had the diagnostic accuracy at 83.3%, sensitivity 75.0%, speciifcity 89.3% and positive predictive value was 83.3%, negative predictive value was 83.3% respectively. FFR and FFRCT showed obvious correlation (r=0.704,P<0.001); Bland-Altman analysis presented good concordance with 95% limits of agreement for FFRCTand FFR value ranged from -0.12 to 0.16, and 95.8% of the points (46/48) fell in the 95% limit of agreement, Receiver operating characteristic curve indicated that AUC of FFRCT was 0.871 (95% CI 0.770-0.973). Conclusion: CTA could accurately assess FFR, and FFRCT might be used in guiding the treatment for patients with intermediate coronary stenosis in clinical practice.
10.Study on expression of RACK1,Src and Bcl-2 protein and their correlation in gastric carcinoma tissues and adjacent carcinomatous tissues
Junmao CHEN ; Siyang LIU ; Jinghua WU ; Jianli CHEN ; Peng ZHAO ; Xiaotao WANG ; Guanghua YANG ; Changyou WANG ; Guozhi ZHANG ; Zhenxing LI
Chongqing Medicine 2016;45(19):2645-2647,2651
Objective To investigate the expression change of RACK1 ,Src and Bcl‐2 in gastric carcinoma tissue and adjacent carcinomatous tissue .Methods Eighty specimens of gastric carcinoma and adjacent carcinomatous tissues in our hospital from Au‐gust 1 ,2011 to February 1 ,2014 were collected .The immunohistochemistry staining and Western blotting methods were adopted to detect the expression of RACK1 ,Src and Bcl‐2 protein in gastric carcinoma and adjacent carcinomatous tissues ,and their correlation was analyzed and performed the statistical analysis by combining with the clinicopathological data .Results The immunohistochem‐istry staining and Western blotting detection displayed that the expression positive rate and expression level of RACK 1 in gastric carcinoma tissue were obviously lower than those in the adjacent carcinomatous tissue ,while the expression positive rate and ex‐pression level of Src and Bcl‐2 in gastric carcinoma tissue were obviously higher than those in the adjacent carcinomatous tissue ,the differences were statistically significant (P<0 .05) .The RACK1 expression in gastric carcinoma tissue was negatively correlated with the Src and Bcl‐2 expression(r= -0 .632 ,-0 .754 ,P<0 .01) ,while Src had no obvious correlation with Bcl‐2 protein(r=0 .217 ,P>0 .05) .Conclusion The expression of RACK1 in gastric carcinoma tissue is significantly decreased ,while the expres‐sions of Src and Bcl‐2 are increased .

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