1.Difluoromethylornithine suppresses growth and improves apoptosis of human leukemia Jurkat cells
Yushan REN ; Yu HAN ; Chunyu CAO ; Yanlin WANG
Journal of Third Military Medical University 1988;0(06):-
Objective To investigate whether difluromethylornithine (DFMO) can be used in the treatment of human leukemia. Methods The cell proliferation was detected by MTS [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium)] assay after treatment of human lymphocyte Jurkat cells by DFMO (0 to 10 mmol/L) for 24 to 72 h. Enzyme activity of spermine oxidase (SMO) and acetylpolyamine oxidase (PAO) was determined by chemiluminesence assay. DNA fragmentation assay was used to evaluate cell apoptosis. Fluorescent dye assay was performed to determine the changes in mitochondrial membrane potential. Western blotting was used to determine Bax content. Casepase-3 enzyme activity was measured by spectrophotometric method. Results DFMO treatment inhibited the proliferation of Jurkat cells significantly in a dosage- and time-dependent manner (P
2.The application of diffusion weighted imaging in the cervical spondylotic myelopathy with 3.0T MRI
Kaiming CAO ; Nanxin HAO ; Yibin WANG ; Genlin ZONG ; Yushan DU
Journal of Practical Radiology 2014;(6):918-922
Objective To explore the value of diffusion weighted imaging (DWI)as a quantitative analysis tool for chronic cervical spondylotic myelopathy (CSM)with 3.0T MRI.Methods Cervical routine MR T2-weighted imaging(T2 WI)and diffusion weighted imaging were performed in 38 patients with cervical spondylotic myelopathy (CSM group)and 20 healthy volunteers (control group),and sagittal image of apparent diffusion coefficient (ADC)was reconstructed.The number of high T2 signal of the spinal cord were calculated in both healthy volunteers and CSM patients.The ADC values of CSM groups were measured in C3,T1 -C2 level and the compression or abnormal signal parts of the spinal cord;the control group measured C3、C4-C6 and T1 -T2 level in the spinal cord.Results There were 3 cases,1 case,6 cases and 8 cases high T2 signals in CSM patients from Ⅰ to Ⅳ levels,while there were 2 cases show high T2 signals in control group,respectively.The mean ADC values of the compression part were gradual-ly increased in CSM group from Ⅰ to Ⅳ level.There were significant differences of the mean ADC values of compression part be-tween the control group,CSMⅠ-Ⅱ level and the CSM Ⅳ level.Conclusion From our study,the change of ADC value of spine cord occured earlier than the change of signal in CSM patients.Therefore,the ADC value of spine cord might reflect the degree of spinal cord compression from CSM patients.
3.Prokaryotic expression and polyclonal antibody preparation of human spermine oxidase
Chunyu CAO ; Yu HAN ; Yushan REN ; Yanlin WANG
Journal of Cellular and Molecular Immunology 2009;25(10):920-923
AIM: To prepare recombinant human spermine oxidase (SMO) and polyclonal antibody against human SMO by gene recombination techniques. METHODS: Human SMO cDNA was amplified from total RNA of A549 cells through reverse transcription PCR. The cDNA was then cloned into pET-15b to construct SMO prokaryotic expression vector. After transforming, the vector was induced to express recombinant SMO by IPTG in E. coli BL21 (DE_3). Recombinant SMO was purified by Ni-NTA resin under denaturing condition and then was dialyzed to renature. The enzyme activity of recombinant SMO was analyzed by chemical fluorescent method. SMO polyclonal antibody was prepared by using recombinant human SMO protein purified by polyacrylamide gel electrophoresis as antigen to inoculate rabbit intradermally. The titer and specificity of anti-sera were determined by ELISA, Western blot and Immune Cell Chemistry. RESULTS: Purified and dialyzed recombinant human SMO has the specificicity of oxidizing the spermine. The polyclonal antibody has high titer and specificity against human SMO. CONCLUSION: This research established a method for prokaryotic expression, purification and polyclonal antibody preparation of human SMO. The method lays a foundation for the future functional research of SMO.
4.Anti-rejection effects of CTLA-4 Ig fusion protein on cardiac allografts in mice and its mechanism in vivo
Guoshan DING ; Quanxing WANG ; Ming ZHANG ; Wenji SUN ; Yushan LIU ; Guoyou CHEN ; Zhiren FU ; Xuetao CAO ;
Academic Journal of Second Military Medical University 1985;0(06):-
Objective: To determine the anti rejection effects of CTLA 4 Ig fusion protein on cardiac allografts in mice and to discuss its mechanism in vivo . Methods: BALB/c recipients were performed cervical heterotopic heart transplantation to receive C57BL/6 donor hearts with a cuff technique. BALB/c recipients were intraperitoneally injected with CTLA 4 Ig [100 ?g/d?15 times], control immunoglobins and PBS to observe the survival time of allografts with ECG. The hyporesponsiveness of splenic T cell, the polarization of the T subsets were analyzed after the recipients treated with CTLA 4 Ig. Results: After treated with CTLA 4 Ig, the survival of cardiac grafts was significantly prolonged compared with the control groups, and more than 40% cardiac grafts survived over 2 months. The splenic T cells isolated from recipients did not respond to restimulation of donor splenocytes in MLR, but did exhibit the capacity to proliferate in response to C3H splenocytes(third party).The levels of IL 2 and IFN ? decreased and the level of IL 10 increased in CTLA 4 Ig treated mice. Conclusion: Administration of CTLA 4 Ig can induce donor specific tolerance, which induce T subsets to polarize toward Th2 subset and hyporesponsiveness to alloantigen, and prolong the survival time of the cardiac grafts effectively. [
5.Effect of Electroacupuncture on the Behavior of APP/PS1 Transgenic Mice of Different Age
Xin WANG ; Anping XU ; Jin CAO ; Menghan LU ; Wei Lü ; Quankai JING ; Yang GAO ; Yushan GAO ; Weiguo XUE ; Zhigang LI
Shanghai Journal of Acupuncture and Moxibustion 2017;36(5):614-619
Objective To study the effect of electroacupuncture on the behavior of APP/PS1 transgenic mice of different age, and to explore the optimal intervention time of electroacupuncture in treating Alzheimer's disease (AD). Method APP/PS1 double-transgenic mice of 4 months, 6 months and 9 months old, 20 in each age group, were randomized into a model group and an electroacupuncture group, and ten C57BL/6 wild-type mice were taken as a control group; after 6-week electroacupuncture treatment, the Morris water maze was adopted for spatial memory and behavioral test, and the changes of behavior in each group were observed.Result Of the 5-month-old mice, there were no significant between-group differences (P>0.05), while the time factor (day) produced a statistical significance (P<0.01); of the 7-month-old mice, there were significant differences considering the different groups, time factor and interaction (P<0.01); of the 10-month-old mice, there were significant differences considering the different groups, time factor and interaction (P<0.05). The spatial probe test showed that there were significant between-group differences in comparing the platform crossings and swimming distance in platform quadrant in each age group (P<0.05).Conclusion Electroacupuncture can improve the learning and memory of APP/PS1 double-transgenic mice, the age of 6-7 months old is possibly the optimal intervention time of electroacupuncture for AD, but there still requires further mechanism studies.
6.Secretory carcinoma of salivary gland: a clinicopathological and prognostic analysis of twelve cases
Chinese Journal of Pathology 2021;50(8):899-903
Objective:To study the clinical manifestations, pathological features, molecular features, differential diagnosis and prognosis of secretory carcinoma of salivary gland (SCSG).Methods:Twelve cases of SCSG diagnosed in the Department of Pathology, the First Affiliated Hospital of Zhengzhou University from January 2017 to December 2019 were collected and analyzed in terms of histological morphology, immunohistochemistry (Envision method) and molecular detection.Results:Among the 12 patients, there were 6 males and 6 females, aged 12-67 years old, with a median age of 41.5 years. The lesions in 11 patients were located in the parotid gland. The maximum diameter of the tumors ranged from 0.8 to 5.5 cm. Clinically, slow-growing painless or tender mass was the first symptom, and all patients underwent surgical resection, among which 5 patients received postoperative radiotherapy and chemotherapy, and 1 patient developed local recurrence. Histopathologically, the tumor is characterized by microcystic, solid or tubular structures with unique intravitary homogeneous secretions. Immunohistochemistry showed diffuse expression of CK7, mammaglobin, GATA-3 and S-100 in all cases, and pan-Trk in 10 of the 12 cases. Sox-10 was expressed focally in 9 cases, and Ki-67 index was 5%-20%. In molecular detection, 11 cases had ETV6 gene break/fusion.Conclusions:SCSG is a relatively rare low-grade malignant salivary gland tumor, with typical histological morphology and immunophenotype, Pan-Trk immunohistochemistry may be related to NTRK fusion, ETV6-NTRK3 gene rearrangement is not only of diagnostic significance, but also Trk-targeted therapy is expected to play a greater role in clinical treatment.
8.Diagnostic value of D-dimer for chronic periprosthetic infection after hip and knee joint replacement
Runqing MIAO ; Li CAO ; Yushan NUERAIJIANG ; Xiaogang ZHANG ; Wulamu WUHUZI ; Jiangdong REN ; Rendong JIANG ; Qian WANG
Chinese Journal of Surgery 2020;58(6):464-468
Objective:To investigate the diagnose value of D-dimer for chronic periprosthetic infection (PJI) after hip and knee arthroplasty.Methods:A retrospective analyze was conducted on 168 patients underwent revision arthroplasty and primary arthroplasty at the First Affiliated Hospital of Xinjiang Medical University from November 2017 to December 2018.There were 58 males and 110 females, aged(58.6±14.5)years.There were 48 cases of chronic PJI (21 cases of knee joint, 27 cases of hip joint), 57 cases of aseptic loosening (16 cases of knee joint, 41 cases of hip joint), and 63 cases of normal follow-up patients after hip (35 cases) or knee (28 cases) arthroplasty.The levels of D-dimer, erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) were collected.The levels of D-dimer in patients with chronic PJI of hip and knee joints were compared by Mann-Whitney U test.The diagnostic efficacy of D-dimer, ESR and CRP in chronic PJI of hip and knee joints was analyzed by receiver operator curve (ROC). Results:The D-dimer level was significantly higher in knee chronic PJI patients than hip chronic PJI patients( M ( QR) ) (1 040 (1 140.5) μg/L vs.435 (605) μg/L, Z=3.169, P=0.002) . ROC analysis showed that the optimum cutoff value of D-dimer in the diagnosis of chronic PJI was 370.5 μg/L, the sensitivity was 90.5%, the specificity was 84.1%; the optimum cutoff value of CRP was 9.3 mg/L, the sensitivity was 95.2%, the specificity was 90.9%, the optimum cutoff value of ESR was 33 mm/h, the sensitivity was 90.5%, and the specificity was 88.6%.The optimum cutoff value of D-dimer in the diagnosis of chronic PJI of hip joint is 294 μg/L, the sensitivity of diagnosis is 66.7%, the specificity is 77.6%; the optimum cutoff value of ESR is 45 mm/h, the sensitivity of diagnosis is 55.6% , the specificity is 97.4%; the optimum cutoff value of CRP is 8.1 mg/L, the sensitivity of diagnosis is 74.1%, the specificity is 84.2%. Conclusion:The value of D-dimer in the diagnosis of chronic PJI of knee joint is higher than that of hip joint, but the value of D-dimer in the diagnosis of chronic PJI is not better than ESR and CRP.
9.Effects of electroacupuncture combined with hydrogel on the formation and changes in the glial scar in rats with spinal cord injury
Yang KAITAN ; Gao YUSHAN ; Yang YONGDONG ; Li CHUANHONG ; Cao ZHENG ; Wang XIUMEI ; Li ZAIBO ; Yu XING
Journal of Traditional Chinese Medical Sciences 2022;9(2):197-205
Objective:To observe the effect of electroacupuncture(EA)combined with oriented conductive bio-protein hydrogel(OCBH)on the recovery of nerve function in rats with complete spinal cord injury(SCI)and to explore its effect and mechanism on the formation and changes of glial scars.Methods:A total of 72 female Sprague-Dawley rats were randomly divided into groups according to the treatment received.A rat model of complete SCI was constructed using a spinal cord transection.Behavioral assessments,hematoxylin-eosin(H&E)staining,immunofluorescence staining,and Western blotting were performed at a fixed period after the operation.Results:The material group and the material+EA group obtained better results in the behavioral as-sessments(all P<.05)and the H&E staining.In the immunofluorescence staining and Western blotting,the GFAP protein was expressed more and denser in the material group and the material+EA group than in the model group,and the density of the GFAP expression in the material+EA group was lower at week 12 than in the material group(all P<.05).The expression of complement C3 in the model,material,and material+EA groups decreased in turn.Some inflammatory factors and the NF-kB signaling pathway showed similar results in the Western blotting(all P<.05).The expression of the GDNF protein in the material+EA group was significantly higher than that in the model group and the material group(both P<.01).Conclusion:EA combined with OCBH can promote the recovery of motor functions after SCI by facili-tating the formation of glial scars in the early stage,preventing the further spread of an inflammatory response that would affect the activation of A1/A2 astrocytes and change the morphology of glial scars at the spinal cord-material interface in its late stage.
10.Gender differences in serological indicators for the diagnosis of chronic periprosthetic infections
Shuaifei TIAN ; Xiaobin GUO ; Wulamu WUHUZI· ; Yushan NUERAIJIANG· ; Yicheng LI ; Xiaogang ZHANG ; Li CAO
Chinese Journal of Orthopaedics 2023;43(11):768-774
Objective:To investigate the gender differences in serum C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), platelet (PLT), fibrinogen (FIB) and D-dimer for the diagnosis of chronic periprosthetic joint infection (PJI).Methods:A total of 470 patients who underwent revision arthroplasty in the Department of Joint Surgery, First Affiliated Hospital of Xinjiang Medical University from January 2013 to December 2021 were retrospectively analyzed. According to gender and diagnosis, they were divided into four groups: 78 cases of male chronic PJI, 108 cases of male aseptic loosening, 95 cases of female chronic PJI, and 189 cases of female aseptic loosening. The serological results of CRP, ESR, PLT, FIB and D-dimer were collected before operation. The receiver operating characteristics (ROC) curves were used to determine the optimal cut-off values of CRP, ESR, PLT, FIB and D-dimer for the diagnosis of chronic PJI in different genders, and to calculate their sensitivity and specificity. The diagnostic efficacy of the index was analyzed by comparing the area under curve (AUC) of different indicators.Results:The levels of ESR, PLT and D-dimer in the male chronic PJI group were 43 (20.0, 52.5) mm/1 h, 249×10 9 (204×10 9, 306×10 9) /L, 449 (219,833) μg/L, respectively, which were lower than those in the female group of 56 (40, 65) mm/1 h, 295×10 9 (228×10 9, 364×10 9) /L, and 645 (345, 1 157) μg/L, with statistically significant differences ( Z=-4.17, P<0.001; Z=-2.17, P=0.030; Z=-2.82, P=0.005). The AUC of CRP in the male chronic PJI group was 0.841, which was higher than the AUC of the other four indicators; CRP was combined with ESR, PLT, FIB and D-dimer to establish a joint prediction model for male chronic PJI. The ROC curve showed that the combination of CRP+FIB had a maximum AUC [0.849, 95% CI (0.79, 0.91)], sensitivity of 80% and specificity of 86%. The AUC of CRP in the female chronic PJI group was 0.866, which was higher than the AUC of the other four indices; CRP was combined with ESR, PLT, FIB and D-dimer to establish a combined prediction model for female chronic PJI. The ROC curve showed that the combination of CRP+PLT had the maximum AUC [0.883, 95% CI (0.84, 0.93)], sensitivity of 87% and specificity of 79%. Conclusion:Serologic indicators in patients with chronic PJI are gender-specific. CRP combined with FIB has the highest diagnostic value for the chronic PJI in males, while CRP combined with PLT has the highest diagnostic value for the chronic PJI in females.