1.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.
2.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.
3.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
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.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.
9.Diagnostic value of plasma fibrinogen in chronic periprosthetic infection
Shuaifei TIAN ; Xiaobin GUO ; Yicheng LI ; Yushan NUERAIJIANG· ; Wulamu WUHUZI· ; Xiaogang ZHANG ; Li CAO
Chinese Journal of Orthopaedics 2023;43(13):891-897
Objective:To investigate the diagnostic value of plasma fibrinogen (FIB) in chronic periprosthetic joint infection (PJI).Methods:A total of 470 patients who underwent revision hip and knee arthroplasty after primary hip and knee arthroplasty in the First Affiliated Hospital of Xinjiang Medical University from January 2013 to December 2021 were retrospectively analyzed. According to the diagnosis of the disease, 173 patients (112 hips and 61 knees) were divided into chronic PJI group, including 78 males and 96 females, aged 65 (53, 72) years; and 297 patients (216 hips and 81 knees) were divided into aseptic loosening group, including 108 males and 189 females, aged 63 (50, 72) years. The preoperative levels of FIB, C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), D-dimer and platelet were compared between the two groups, and the sensitivity and specificity of the diagnosis of chronic PJI were calculated. Receiver operating characteristic (ROC) curves were used to determine the optimal cut-off values of FIB, CRP, ESR, platelet and D-dimer for the diagnosis of chronic PJI. The diagnostic efficacy of each indicator was analyzed by comparing the area under curve (AUC) and using a combined diagnostic test.Results:FIB, CRP, ESR, platelets and D-dimer in the chronic PJI group were 4.05 (3.52, 4.72) g/L, 19.5 (10.7, 40.0) mg/L, 50 (28, 60) mm/1 h, 270 (221, 351)×10 9 /L, 514 (261, 873) μg/L, respectively, which were higher than 3.25 (2.80, 3.63) g/L, 3.7 (2.0, 6.7) mg/L, 20 (12, 30) mm/1 h, 225 (182, 269)×10 9 /L, and 310 (167, 569) μg/L in sterile loosening group, with statistically significant differences ( P<0.05). The AUC of FIB, CRP, ESR, platelets and D-dimer in the diagnosis of chronic PJI were 0.78 (95% CI: 0.73, 0.82), 0.86 (95% CI: 0.82, 0.89), 0.80 (95% CI: 0.76, 0.85), 0.68 (95% CI: 0.63, 0.73), 0.64 (95% CI: 0.59, 0.69); the optimal cut-off values were 3.73 g/L, 9.64 mg/L, 39 mm/1 h, 280×10 9 /L, 624 μg/L; the sensitivity was 68%, 79%, 69%, 47%, 43%; the specificity was 81%, 85%, 85%, 81%, 79%, respectively. When CRP, ESR, FIB, platelets and D-dimer were combined sequentially to diagnose patients with chronic PJI, the sensitivity and specificity of the series test were 12.7% and 99.7%, and those of the parallel test were 100% and 37.3%. The combined diagnostic test showed that the maximum AUC of FIB combined with CRP was 0.85 (95% CI: 0.81, 0.89), with a sensitivity of 76% and a specificity of 89%. Conclusion:The clinical value of plasma FIB in the diagnosis of chronic PJI is not superior to that of CRP and ESR, but the combination of FIB and CRP can improve the specificity.
10.Analysis of bacterial spectrum and variability of drug resistance of bile in patients with common bile duct stones and biliary tract infections
Hang ZHANG ; Peng QI ; Cong XIE ; Yushan MENG ; Kuijin XUE ; Lu LIU ; Guangrong WANG ; Wanting LIU ; Baoguo HE ; Hui JU ; Bin CAO
Chinese Journal of Digestion 2023;43(1):24-30
Objective:To analyze the distribution of pathogenic bacteria in bile culture in patients with common bile duct stones and biliary tract infections, in order to guide clinical optimization of antibiotics application.Methods:From March 30, 2017 to December 31, 2021, at Affiliated Hospital of Qingdao University, 753 patients with common bile duct stones and biliary tract infections and received endoscopic retrograde cholangiopancreatography were selected. Bile samples were obtained for bacterial culture, strain type identification and drug sensitivity test in order to analyze bile pathogenic bacteria distribution, change trend and drug resistance. Chi-square test was used for statistical analysis.Results:From 2017 to 2021, the total positive rate of bile culture in 753 patients with choledocholithiasis complicated with biliary tract infection was 90.17% (679/753). From 2017 to 2021, the positive rates of bile culture were 82.05% (64/78), 88.81% (119/134), 88.03% (125/142), 93.87% (199/212), and 91.98% (172/187), respectively, and the difference was statistically significant ( χ2=10.78, P=0.029). The positive rate of bile culture in 2017 was lower than those in 2020 and 2021, and the differences were statistically significant ( χ2=9.43 and 5.57, P=0.002 and 0.018). There were no significant differences in the positive rates of bile culture among the other years (all P>0.05). A total of 1 033 pathogenic bacteria were detected in the 679 bile specimens with positive bile culture results. Among which the total proportion of Gram-negative bacilli was 57.02% (589/1 033), and from 2017 to 2021 the proportions were 66.38% (77/116), 66.47% (111/167), 59.43% (104/175), 54.75% (173/316), and 47.88% (124/259), respectively. The total proportion of Gram-positive cocci was 41.05% (424/1 033), and from 2017 to 2021 the proportions were 31.90% (37/116), 31.74% (53/167), 38.86% (68/175), 44.30% (140/316), and 48.65% (126/259), respectively. The total proportion of fungus was 1.94% (20/1 033), and from 2017 to 2021 the proportions were 1.72% (2/116), 1.80% (3/167), 1.71% (3/175), 0.95% (3/316), and 3.47% (9/259), respectively. From 2017 to 2021, the proportion of Gram-negative bacilli gradually decreased, while the proportion of Gram-positive cocci gradually increased, and the differences were statistically significant ( χ2=20.14 and 17.91, P<0.001 and =0.001). From 2017 to 2021, the change in the proportion of fungus was not statistically significant ( P>0.05). The main Gram-negative bacilli in the bile culture were Escherichia coli (31.36%, 324/1 033) and Klebsiella pneumoniae (12.68%, 131/1 033); the main Gram-positive cocci were Enterococcus faecalis (14.04%, 145/1 033) and Streptococcus salivarius (4.36%, 45/1 033). From 2017 to 2021, the proportions of Escherichia coli were 39.66% (46/116), 38.92% (65/167), 33.14% (58/175), 28.48% (90/316), and 25.10% (65/259), respectively, with gradual decrease and the difference was statistically significant ( χ2=14.34, P=0.006). From 2017 to 2021 the detection rates of extended-spectrum β-lactamase (ESBL) in Escherichia coli and Klebsiella pneumoniae were 30.43% (14/46), 26.15% (17/65), 29.31% (17/58), 38.89% (35/90), 40.00% (26/65), and 4/15, 20.00% (5/25), 20% (5/25), 24.32% (9/37), and 31.03% (9/29), and there were no significant differences in the detection rates of ESBL between different years (both P>0.05). Conclusions:From 2017 to 2021, the positive rate of bile culture in patients with choledocholithiasis complicated with biliary tract infection showed an overall increasing trend. Gram-negative bacilli were still dominated in bile pathogenic bacteria, while the proportion of Gram-positive cocci remarkably increased, and the bile bacterial spectrum significantly changed. Clinicians should adjust the antibiotic dosing regimens according to the variation of bacterial spectrum and drug resistance.