1.Inhibitory effect of transgiutaminase 2 on apoptosis of osteosarcoma cell line MG-63 and its mechanism
Wentao CAI ; Hong XIA ; Ningjiang SHENG ; Mingxia LIN
Journal of Jilin University(Medicine Edition) 2014;(4):782-789
Objective To investigate the anti-apoptosis effect of transgiutaminase 2 (TG2)in osteosarcoma cell line MG-63 and to explore the mechanism of inhibiting apoptosis of tumor cells.Methods The TG2-tgrgeted siRNA was designed,and the hypoxia culture model of MG-63 cells was established by a hypoxia incubator and the cells were divided into four groups:normal oxygen group,the cells were cultured under normal oxygen;hypoxia group, the cells were cultured in hypoxic incubators;control siRNA hypoxia group,the cells were cultured in hypoxic incubators after transfection of control siRNA;TG2 siRNA hypoxia group , the cells were cultured in hypoxic incubators after transfection of TG2 siRNA.The expressions of Bax and cytochrome C (Cyt C)and the apoptotic rates were observed at different time (6,12,24,48,and 72 h)after hypoxia culture.Microtiter plate assay was performed to monitor the intracellular TG2 activity.RT-PCR was used to detect the mRNA expressions of TG2 and Bax.The expressions levels of TG2,Bax and Cyt C were observed by immunohistochemical staining and Western blotting method.The apoptotic rates were analyzed using flow cytometry.Results Compared with normal oxygen group,the activity of TG2,the mRNA and protein expression levels of TG2 in hypoxia group and control siRNA hypoxia group were increased remarkably with the prolongation of the hypoxia time (P<0.01);the expression level of Bax protein was decreased significantly (P<0.01 ), but the expression level of Bax mRNA had no significant change (P>0.05);the expression levels of Cyt C protein in cytoplasm and mitochondria and the apoptotic rates had no markedly changes (P>0.05).Compared with hypoxia group and control siRNA hypoxia group,the expression levels of mRNA and protein of TG2 in TG2 siRNA hypoxia group were decreased significantly at different time points (P<0.01);the protein expression levels of Bax and Cyt C in cytoplasm and the apoptotic rates were increased markedly (P<0.01 );the expression level of Cyt C in mitochondria was decreased (P<0.01).Conclusion TG2 can inhibit the apoptosis of tumor cells through down-regulating the Bax expression and preventing Cyt C releasing into the cytoplasm.
2.Expression of recombinant human tumor suppressor NDPK-A in E. coli
Yanchao RAN ; Yifei WANG ; Sheng XIONG ; Meiying ZHANG ; Wentao HUANG ; Linbo LUO ; Qiuying LIU
Chinese Journal of Pathophysiology 1989;0(06):-
AIM: To construct E. coli expression plasmid of recombinant human NDPK-A with a 6?His tag, optimize the expression condition and identify the activity of the product. METHODS: nm23-H1 was subcloned from plasmid pBVNMH1 to pQE40 which contain 6?His purification tag. The expression condition was modulated in grades to get the optimal expression. We purified protein with the Ni+-NTA affinity chromatography column, identified the immunogenicity of the product with Western blot, and measured the kinases activity with HPLC. In addition, angiogenesis inhibition activity of rhNDPK was identified by CAM. RESULTS: The sequence of nm23-H1 subclone in pQE40 was exactly correct. The expression rate of rhNDPK-A was 49 6%. Purified rhNDPK-A specially recognized the antiserum of NDPK-A. It also inhibited angiogenesis. CONCLUSION: PQE-nm23H1 containing 6?His can express target protein at high level. This purification method is simple than other methods, and the product has the same activity as natural human NDPK-A.
3.Interpretation for MRI signal evolution of hepatic tumors after microwave ablation
Zhenyu JIA ; Qifeng CHEN ; Wentao WU ; Sheng LIU ; Haibin SHI ; Zhengqiang YANG
Journal of Interventional Radiology 2017;26(4):324-328
Objective To make an interpretation for the time-related evolving process of magnetic resonance imaging (MRI) signal of hepatic tumors after microwave ablation (MWA) treatment.Methods A total of 56 patients with malignant hepatic tumors (56 lesions in total) were enrolled in this study.Upper abdominal MRI plain scan and enhanced scan were performed in all patients at the second day,one month and 6 months after MWA treatment.The MRI signal features of ablation zones at different time points on T1WI,T2WI,DWI as well as on contrast-enhanced T1WI were documented,and the judgment of whether there was tumor recurrence was made.Results Two days after MWA,the ablation zone was manifested as target-like structure on T1WI and T2WI,which was characterized by central high signal ablation zone with low signal band around on T1WI and low signal ablation zone surrounded by high signal band on T2WI.One and 6 months after MWA,the volume of ablation area was atrophied,the target-like structure could still be observed on T1WI and T2WI,and the signal of ablation zone became intensified.Contrast-enhanced MRI revealed that abnormal high perfusion sign could be observed around the ablation zone,and on MRI scans performed at two days,one and 6 months after MWA,the ablation zone showed no enhancement.DWI indicated that two days after MWA the signal around the ablation zone was heightened,which decreased gradually in one and 6 months after MWA.Conclusion The signal of the ablation zone of hepatic tumor after MWA is evolving over time.Correct interpretation of MRI signal of ablation zone is helpful for the judgment of curative effect and for the making of therapeutic plan.
4.Holmium laser coagulation under choledochoscope for the treatment of intrahepatic hemobilia
Ruiwu DAI ; Yong YAN ; Bingyin ZHANG ; Danqing LIU ; Wentao BAI ; Sheng ZHANG ; Lijun TANG
Chinese Journal of Digestive Surgery 2013;(6):452-454
Objective To investigate the efficacy of holmium laser coagulation under choledochoscope for the treatment of intrahepatic hemobilia.Methods The clinical data of 55 patients with intrahepatic hemobilia who received holmium laser coagulation under choledochoscope at the General Hospital of Chengdu Military Area from June 2003 to August 2011 were retrospectively analyzed.After the hemobilia was confirmed with choledochoscopy,an optical fiber of holmium laser was inserted into the intrahepatic bile duct from the hole of choledochoscope.The top of the optical fiber was posed close to the bleeding region,and then the laser was stimulated for coagulation under direct vision.If the observation of the bleeding area was influenced by the bleeding,the porta hepatis was temporarily occluded with a rubber hose combined with pressurized liquid injection bag.For patients with distal bile duct stenosis and bleeding,the narrow area was first expanded by biliary balloon or laser,then hemostasis was completed after the flow of bile duct was restored.Results The hemorrhage of the 55 patients was controlled,with an average time of 5.5 minutes (range,2-15 minutes).Temporary hepatic portal clamping was applied for 16 patients (27 times),and the average blocking time was 2.2 minutes (range,1-4 minutes).Biliary rehaemorrahagia occurred after operation in 2 patients,and they were cured by non-surgical treatment.All patients were discharged after successful hemostasis.The time of follow-up was 35-49 days.The epithelium of the bile duct coagulated by intraoperative holmium laser restored flat in 41 patients,and new biliary stricture was not found in the other 14 patients.Conclusion Holmium laser coagulation under choledochoscope for intrahepatic hemobilia is simple and effective.
5.Changes of Tim-3 expression In T lymphocytes from different sites in mice heart-transplant recipients
Zemin FANG ; Wentao HE ; Sheng WANG ; Lanjun CAI ; Zhenlong LUO ; Weina ZHANG ; Hongmin ZHOU ; Zhonghua CHEN ; Changsheng MING
Chinese Journal of Organ Transplantation 2010;31(3):141-143
Objective To explore the expression level of Tim-3,the marker of activated T_H 1 cells.in T lymphocytes in different sites from recipients with acute rejection.Methods The model of cervical heterotopic heart transplantation was established in mice Two groups were get up:the isograft group(C57BL/6→C57BL/6) and the allograft group (Balb/c→C57BL/6).Lymphocytes were isolated from peripheral blood,spleens,draining lymph nodes and grafts 3 or 6 days after transplantation.The expression of TIM-3 in CD4~+ and CD8~+ T subsets was detected by flow cytometry.Results There was no significant difference in Tim-3~+/CD4~+ and Tim-3~+/CD8~+ ratio in peripheral blood or spleens between two groups.As compared with the isograft group,the proportion of Tirn-3~+/CD4~+ cells was slightly elevated in draining lymph node(P<0.05),but the percentage of Tim-3~+/CD4~+ cells had no significant change between 3 days and 6 days in the allograft group(P>0.05).The expression of Tim-3 in CD4~+ and CD8~+ of graft infiltrating T cells was obviously increased in allograft group(P<0.01),and it was significantly (P<0.01) up-regulated on the 6th day as compared with that on the 3rd day.Conclusion The dynamic changes of Tim-3 expression in T lymphocytes in draining lymph node and graft were correlated with the progresston oi acute rejection in mice.
6.Observation of the cavum septi pellucidum with MRI
Wentao WANG ; Chuqing ZHAO ; Shaoyong GONG ; Sheng QIU ; Xiangfei CHEN ; Bosen WU ; Kai WANG ; Junping WANG ; Leitao WEN ; Xiaoqun YAO ; Guangfu YANG
Journal of Practical Radiology 2015;(8):1239-1242
Objective To study the development characteristics of cavum sepit pellucidun (CSP)in prematures,neonates,infants and adults with MRI.Methods Brain MR images of different subjects including 141 prematures,106 neonates,171 infants and 35 046 adults were observed to determine the incidence and shape of CSP,and to measure its transverse diameter.Results CSP incidences were 100% (141/141)in prematures,97.17% (103/106)in neonates,2.26%(4/177)in infants and 0.82% (287/35 046)in adults respectively,and the CSP was cylinder (44.00%)or triangle in shape (56.00%)in prematures,triangle (76.40%)or fissure in shape (23.60%)in neonates.For infants or adults,each shape accounted for about a third of three kinds of shape respectively.Its mean transverse diameters were 5.7 mm in prematures,4.1 mm in neonates,13.3 mm in infants and 14.3 mm in adults respectivity.Conclusion CSP has different performances at development periods in human being brain.Most close after birth,while fewer remain in the whole life.
7.Analysis of clinicopathological features and risk factors for postoperative complications in the elderly gastric cancer patients.
Sheng LU ; Min YAN ; Chen LI ; Chao YAN ; Xuexin YAO ; Minming CHEN ; Runhua FENG ; Renda BI ; Wentao LIU ; Zhenglun ZHU ; Zhenggang ZHU
Chinese Journal of Gastrointestinal Surgery 2016;19(5):514-521
OBJECTIVETo investigate the clinicopathological features and postoperative short-term complications in the elderly gastric cancer patients.
METHODSClinical data of 270 elderly patients with gastric cancer who underwent gastrectomy in Department of Surgery, Ruijin Hospital, Shanghai Jiaotong University School of Medicine between July 2012 and June 2014 were analyzed retrospectively. Among 270 patients, 220 were 70 to 79 years old( old group) and 50 were ≥80 years old(oldest group). The clinicopathological features were compared between the two groups. Perioperative factors were analyzed to determine if they are associated with postoperative complications. Multivariate logistic regression model was performed.
RESULTSBefore operation, most elderly patients (n=161, 59.6%) had 2 or more than 2 comorbidities, including hypertension (n=154, 57.0%), anaemia (n=126, 46.7%), diabetes (n=53, 19.6%), arhythmia (n=52, 19.3%), cardiovascular disease(n=33, 12.2%), and chronic pulmonary disease(n=28, 10.4%). Elderly gastric cancers were more likely to locate at the lower third of the stomach (n=116, 43.0%). The pathological type was mainly the poorly differentiated carcinoma (n=152, 56.3%), and stage III was more common in TNM staging(n=138, 51.1%). As compared to the old group, the oldest group had more preoperative comorbid diseases(P=0.048), more previous surgery(P=0.029), more preoperative transfusion (P=0.019), more combined cholecystectomy (P=0.007) and feeding jejunostomy (P=0.037), but less tumor invasion of nerves(P=0.045). No significant differences in other clinicopathological parameters were found between the two groups (all P>0.05). A total of 121 (44.8%) patients presented postoperative complications, including severe complication in 30 cases(11.1%) and death in 4 cases(1.5%). Forty-seven patients(17.4%) presented operation-associated complications, including infection in 28 cases(10.4%) and leakage in 21 cases(7.8%). One hundred and seven(39.6%) patients presented non-operation-associated complications, including pneumonia in 48 cases(17.8%), hypertension in 23 cases(8.5%), and arhythmia in 17 cases(6.3%). Postoperative morbidities of Clavien-Dindo class II complication and non-operation-associated complication were higher in the oldest group compared with old group(P<0.05), while other postoperative complications were compared between the two groups, only urinary tract infection was significantly different(P<0.05). Univariate analysis showed that postoperative complications were significantly associated with age(χ(2)=7.308, P=0.007), number of comorbid diseases (χ(2)=10.872, P=0.001), cardiovascular disease (χ(2)=9.412, P=0.002), hypertension (χ(2)=4.934, P=0.026) and preoperative transfusion (χ(2)=3.911, P=0.048). Multivariate analysis showed that only the number of comorbid diseases was an independent risk factor for postoperative complications(OR=2.810, 95% CI: 1.710 to 4.616, P=0.000).
CONCLUSIONNon-operation-associated postoperative complications are more likely to occur in the elderly patients due to more comorbid diseases. Perioperative intensive care should be carried out for the elderly gastric cancer patients with comorbid diseases in order to improve surgical safety and efficacy.
Aged ; Aged, 80 and over ; China ; Comorbidity ; Gastrectomy ; adverse effects ; Humans ; Hypertension ; complications ; Logistic Models ; Multivariate Analysis ; Neoplasm Staging ; Postoperative Complications ; Retrospective Studies ; Risk Factors ; Stomach Neoplasms ; complications ; surgery
8.The diagnostic value of both mammography and MRI in combination with clinical features in high-risk breast lesions
Chao YOU ; Weijun PENG ; Yajia GU ; Sheng CHEN ; Xiaohang LIU ; Tingting JIANG ; Wentao YANG
Chinese Journal of Radiology 2020;54(3):203-208
Objective:To evaluate the value of mammography and MRI combined with clinical features in predicting upgrade to malignancy in high-risk breast lesions.Methods:Data from 230 patients who were diagnosed with high-risk breast lesions and underwent both mammography and MRI examinations before biopsy were analyzed retrospectively from Jan 2017 to Mar 2018 in Fudan University Shanghai Cancer Hospital. The imaging features of both mammography and MRI were analyzed, and the association between mammography, MRI and clinical features were evaluated using pathology as the gold standard. Independent t test and χ 2 test were used to compare the difference of clinical and imaging features between upgrade and non-upgrade groups, using receiver operating characteristic (ROC) curve to test the diagnostic value between mammography and MRI. Binary logistic regression was used to evaluate the correlation between upgrade and clinical, imaging findings. Results:Two hundred and thirty patients had 230 lesions, and 47 cases had atypia upgrade to malignancy during second surgery (upgrade rate was 20.4%). There were statistically significant differences in age, maximum diameter of lesion, and menopausal status between the upgraded and non-upgraded groups ( P<0.05). There was no statistically significant difference in mammographic features between two groups ( P>0.05), while there was statistically significant difference in breast MRI features and background parenchymal enhancement ( P<0.05). For the diagnostic value in predicting upgrade of high-risk lesions, MRI was better than mammography (the areas under ROC curve were 0.913 and 0.606, Z=6.919, P<0.01). Single factor analysis showed that age, lesion size, menopausal status, MRI negative and background parenchymal enhancement on MRI were significantly different for upgrade to malignancy ( P<0.05). Multiple factors analysis showed age and background parenchymal enhancement on MRI were independent factors for predicting upgrade ( P<0.01). Conclusion:For the upgrade to malignancy in high-risk lesions, the diagnostic value of MRI is better than mammography. The elder age and moderate or marked background parenchymal enhancement on MRI may serve as useful predictors of upgrade.
9.Study on the evaluation of glenoid bone defects by MRI three-dimensional reconstruction.
Fei ZHANG ; Lin XU ; Baoxiang ZHANG ; Shoulong SONG ; Xianhao SHENG ; Wentao XIONG ; Ziran WANG ; Weixiong LIAO ; Qiang ZHANG
Chinese Journal of Reparative and Reconstructive Surgery 2023;37(5):551-555
OBJECTIVE:
To investigate the feasibility of MRI three-dimensional (3D) reconstruction model in quantifying glenoid bone defect by comparing with CT 3D reconstruction model measurement.
METHODS:
Forty patients with shoulder anterior dislocation who met the selection criteria between December 2021 and December 2022 were admitted as study participants. There were 34 males and 6 females with an average age of 24.8 years (range, 19-32 years). The injury caused by sports injury in 29 cases and collision injury in 6 cases, and 5 cases had no obvious inducement. The time from injury to admission ranged from 4 to 72 months (mean, 28.5 months). CT and MRI were performed on the patients' shoulder joints, and a semi-automatic segmentation of the images was done with 3D slicer software to construct a glenoid model. The length of the glenoid bone defect was measured on the models by 2 physicians. The intra-group correlation coefficient ( ICC) was used to evaluate the consistency between the 2 physicians, and Bland-Altman plots were constructed to evaluate the consistency between the 2 methods.
RESULTS:
The length of the glenoid bone defects measured on MRI 3D reconstruction model was (3.83±1.36) mm/4.00 (0.58, 6.13) mm for physician 1 and (3.91±1.20) mm/3.86 (1.39, 5.96) mm for physician 2. The length of the glenoid bone defects measured on CT 3D reconstruction model was (3.81±1.38) mm/3.80 (0.60, 6.02) mm for physician 1 and (3.99±1.19) mm/4.00 (1.68, 6.38) mm for physician 2. ICC and Bland-Altman plot analysis showed good consistency. The ICC between the 2 physicians based on MRI and CT 3D reconstruction model measurements were 0.73 [95% CI (0.54, 0.85)] and 0.80 [95% CI (0.65, 0.89)], respectively. The 95% CI of the difference between the two measurements of physicians 1 and 2 were (-0.46, 0.49) and (-0.68, 0.53), respectively.
CONCLUSION
The measurement of glenoid bone defect based on MRI 3D reconstruction model is consistent with that based on CT 3D reconstruction model. MRI can be used instead of CT to measure glenoid bone defects in clinic, and the soft tissue of shoulder joint can be observed comprehensively while reducing radiation.
Male
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Female
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Humans
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Young Adult
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Adult
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Imaging, Three-Dimensional/methods*
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Tomography, X-Ray Computed/methods*
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Joint Instability
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Shoulder Joint/diagnostic imaging*
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Shoulder Dislocation
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Magnetic Resonance Imaging/methods*
10. Research of TLD calibrated in different standard X-ray RQR radiation field in terms of eye lens dose H p(3)
Wenyan LI ; Guiying ZHANG ; Zhichao WANG ; Zeqin GUO ; Mengxiao KANG ; Chuanye LIU ; Wentao SHENG ; Yue ZENG ; Xi LUO ; Linlin YUE
Chinese Journal of Radiological Medicine and Protection 2020;40(1):42-46
Objective:
To compare the calibration result of standard X-ray RQR radiation field between SSDL (NIRP) and CEA LIST LNHB(France), and to explore the feasibility of calibrating