1.CdCl2 induces autophagy and apoptosis in HEK293 cells
Xuanxuan LIU ; Wenqian WANG ; Weiping MAO
Chinese Journal of Pharmacology and Toxicology 2016;30(5):569-575
OBJECTIVE To evaluate the possibility that CdCl2 induces autophagy and apoptosis in HEK293 cells,and the role of extracellular regulated protein kinases (ERK1/2) and AKT proteins in autophagy. METHODS Green fluorescence protein(GFP)-light chain 3B(LC3B)expression plasmid was transfected into HEK293 cells. After 24 h,HEK293 cells were induced with CdCl2 2,4,8 and 10μmol·L-1 for 12 h. The expression of GFP-LC3B was detected by fluorescent microscopy. HEK293 cells were induced with CdCl2 2,4,8 and 10μmol · L-1 without transfection of GFP-LC3B for 12 h while autophagic vacuoles were observed by transmission electron microscopy. The expression of LC3B-Ⅱ/Ⅰproteins and the phosphorylation levels of ERK1/2 and AKT were analyzed by Western blotting. Apoptosis was detected by flow cytometry microscopy. HEK293 cells were treated with 3-MA 20μmol · L-1+CdCl2 10 μmol · L-1 for 12 h before cleaved caspase 3 protein was detected by Western blotting. RESULTS When HEK293 cells were exposed to CdCl2(≤10μmol · L-1)for 12 h,cytoplasmic GFP-LC3B punctuates were observed under the fluorescence microscope,and autophagic vacuoles were observed under an electron microscope. The expression of LC3B-Ⅱ/Ⅰ,p-ERK1/2 and p-AKT proteins was significantly increased in CdCl2-induced cells(P<0.05,P<0.01). Moreover,apoptosis was observed. The addition of 3-MA 20μmol · L-1+CdCl2 10μmol · L-1 enhanced apoptosis. Cleaved capase 3 protein expression was significantly increased(P<0.01). CONCLUSION CdCl2(≤10μmol·L-1)can induce autophagy in HEK293 cells. ERK1/2 and AKT proteins might be associated with the activation of autophagy that is accompanied by apoptosis,suggesting that autophagy can inhibit apoptosis at certain concentrations of CdCl2.
2.The value of PET/CT in the diagnosis of early cervical cancer with retroperitoneal lymph node metastasis
Zhijun HU ; Kaijiang LIU ; Qing LIU ; Hong ZHU ; Peiquan LI ; Yuxin SUN ; Xuanxuan ZHAO
Journal of Chinese Physician 2021;23(2):193-197,202
Objective:To investigate the value of positron emission tomography/computed tomography (PET/CT) in diagnosis of retroperitoneal lymph node metastasis for early cervical cancer.Methods:Retrospectively analyze the preoperative PET/CT examination results and postoperative pathological results of patients with early cervical cancer who underwent surgical treatment from May 5, 2019 to August 31, 2020, and analyze the clinical characteristics, so as to explore the high risk factors of PET/CT in the diagnosis of retroperitoneal lymph node metastasis.Results:The accuracy, sensitivity, specificity and area under the curve (AUC) of PET/CT in the diagnosis of retroperitoneal lymph node metastasis were 75.2%, 60.0%, 81.3% and 0.707, respectively. Univariate analysis showed that 2009 International Federation of Gynecology and Obstetrics (FIGO) stage, tumor diameter >4 cm, lymphovascular space involvement (LVSI) positive, depth of invasion, high squamous cell carcinoma antigen (SCC-Ag) level and cytological grade were important factors for PET/CT in the diagnosis of retroperitoneal lymph node metastasis ( P<0.05); Multivariate analysis showed that tumor diameter >4 cm was an independent risk factor for PET/CT diagnosis of retroperitoneal lymph node metastasis ( P=0.015). Conclusions:PET/CT has a certain diagnostic value in the evaluation of retroperitoneal lymph node metastasis, but it can not fully reflect the facticity of lymph node metastasis; tumor diameter > 4cm is an independent risk factor for PET/CT in the diagnosis of retroperitoneal lymph node metastasis.
3.Double-layer patch containing gestodene and ethinylestradiol :preparation and in vitro/in vivo characterization
Yanli GAO ; Yan XIAO ; Jinying LIANG ; Xuanxuan ZHU ; Ying LI ; Jianping LIU
Journal of China Pharmaceutical University 2009;40(5):416-420
Aim: To prepare a novel contraceptive patch containing gestodene(GEST) and ethinylestradiol(EE), and to study the in vitro characterization and in vivo contraceptive effect. Methods: Double-layer technique was applied to sustain steady 7-day permeation flux of drugs. Polariscope examination was carried out to observe the drug distribution behavior in the patch. Uniformity, adhesion, skin irritation, release and permeation tests were conducted to evaluate in vitro characterization. Anti-implantation and anti-fertility experiments were carried out to investigate its contraceptive effect. Results: The in vitro release profiles of both drugs were in accordance with Higuchi equation. The daily permeated amount of GEST per 10 cm2 patch was about 75 μg while the amount of EE was 30 μg.The in vitro transdermal permeation of both drugs from the patches displayed a zero-order process. Permeation rate constants were 0. 377 μg/(cm2·h) for GEST, and 0. 092 μg/(cm2· h) for EE, respec-tively. After transdermal administration, the embryonic number of the test groups was zero, and the uterus coefficients of those groups were significantly reduced compared with those of the control group (P < 0. 01). Conclusion: Double-layer transdermal drug delivery system(TDDS) could allow the steady 7-day permeation flux of drugs when the drug ratio between the immediate-release layer and the reservoir layer was 1:4. In vivo charac-terization demonstrated its contraceptive effects. The prepared novel patch might be a promising non-oral contra-ceptive preparation.
4.Delay aversion test between the male children with obsessive-compulsive disorder and ones with attention-deficit/hyperactivity disorder
Yuncheng ZHU ; Xixi JIANG ; Xuanxuan CHEN ; Li LIU ; Li ZHANG ; Aiai CAO ; Kang JU ; Xinxin LI ; Guohai LI ; Weidong JI
Chinese Mental Health Journal 2015;(4):273-278
Objective:To clarify the characteristics and differences between obsessive-compulsive disorder (OCD)and attention-deficit/hyperactivity disorder (ADHD)in male children.Methods:Forty boys with OCD and 40 boys with ADHD were selected from out-patient department,and 40 normal boys matched with age,gender and IQ were recruited as the controls.Patients'diagnosis was made according to the International Statistical Classifica-tion of Diseases and Related Health Problems,Tenth Revision (ICD-10).The choice delay tasks (CDT1 /2)were assessed respectively for the measurements of normal or high ecological validity of delay aversion.In CDT1 /2, waiting time and scores of the tasks were indicators reflecting the delay aversion.Results:The scores of CDTs were higher in children with OCD than in the controls [CDT1,(30.9 ±3.2)vs.(27.6 ±3.8);CDT2,(31.3 ±4.4)vs. (28.5 ±4.9);P <0.01].The scores of CDTs were lower in children with ADHD than in the controls [CDT1, [(25.0 ±4.3)vs.(27.6 ±3.8)];CDT2,[(26.2 ±4.6)vs.(28.5 ±4.9);P <0.001].Conclusion:The results in-dicate that the children with OCD have a trend to choose the task with longer waiting time and higher score,and the children with ADHD trend to choose the task with shorter waiting time and lower score.The results suggest that the pathogenesis of hyperactivity/impulse control may be different between OCD and ADHD in delay aversion.
5.Application of shear wave elastography in cervical cancer
Manting ZENG ; Jihua LIU ; Ningbo ZHOU ; Jian WANG ; Xuanxuan LI ; Hong ZHU
Journal of International Oncology 2019;46(2):117-120
Shear wave elastography (SWE) is used to quantitatively analyze the hardness of the tissue by Young's modulus.The hardness of the tissue is visualized in the form of color coding to distinguish the benign and malignant tissue detected.SWE has higher sensitivity,accuracy and specificity compared with traditional color doppler,which is more objective than elastography,safer,cheaper and simpler than MRI.SWE has a good application prospect in the diagnosis,clinical staging and curative effect monitoring of cervical cancer.
6.Advances in the effect of body temperature on perioperative neurocognitive disorders in elderly patients
Chinese Journal of Geriatrics 2023;42(2):225-228
Perioperative neurocognitive disorders(PND)is a common complication after surgery or anesthesia, especially in elderly patients.The effects and mechanism of body temperature on PND are still controversial.In this paper, the relationship between body temperature and PND and its possible mechanism of action were discussed based on some research results of perioperative body temperature management in elderly patients, aiming at suggesting the significance of body temperature management in preventing PND.
7.Research progress on cervical cerclage for preventing preterm birth in twin pregnancies
Peili WU ; Qing XUE ; Xuanxuan LIU ; Xiao SUN
Chinese Journal of Perinatal Medicine 2024;27(3):258-261
Preterm birth is the most common maternal complication in twin pregnancies. In recent years, cervical cerclage has been of long-standing interest in the prevention of preterm birth in twin pregnancies. However, its clinical application in the treatment of cervical insufficiency of twin pregnancies remains a controversial subject. In addition, infection or inflammation conditions are considered to be closely related to the perinatal outcomes of twin pregnancies after cervical cerclage. This paper reviews the research progress on cervical cerclage in twin pregnancies, recommending cervical cerclage for twin pregnancies with cervical length≤15 mm or cervical dilatation, while it is not suggested for those with cervical length of 15-25 mm or history-indicated cervical cerclage. The clinical significance of preoperative evaluation of intraamniotic infection or inflammation of twin pregnancies needs to be further explored, but it is necessary to avoid the effect of antibiotic use on the evaluation of surgical effects.
8.Application values of shear wave elastography and ultrasonic grading in liver grafts from brain death donor
Xuanxuan LI ; Wenyi LIU ; Jianhong WANG ; Yang ZHAO ; Lina ZHU ; Xin WANG ; Yuan GUO ; Yong LIU
Chinese Journal of Organ Transplantation 2019;40(4):211-214
Objective To explore the values of shear wave elastography (SWE) and ultrasonic grading in liver grafts from brain death donor (DBD).Methods Liver grafts of 39 DBD cases were examined preoperatively by ultrasonography (US) and SWE.Blood samples were tested preoperatively.Based upon early allograft function of liver recipients,the donors were classified into early allograft dysfunction (EAD) and early allograft function normal (non-EAD) groups.The SWE/ US grading results of EAD group were compared with those of non-EAD group.Receiver operating curve (ROC) was employed for analyzing the diagnostic accuracy of SWE/US grading in EAD.Results The SWE value of EAD group was significantly higher than that of non-EAD group [(6.65 ± 2.69)vs.(3.50 ± 1.27) kPa,P<0.05].Ultrasonic grading of EAD group was also significantly higher than that of non-EAD group (P<0.05).The area under ROC curve (AUROC) of SWE in EAD was 0.939,optimal cut-off value 4.56 kPa,AUROC of ultrasonic grading 0.806 (P =0.003) and optimal cut-off value level 3.5.Conclusions SWE quantifies fibrosis in liver grafts of DBD with a high diagnostic accuracy.There are significant correlations between EAD after liver transplantation and SWE value of liver grafts and ultrasonic grading.SWE and ultrasonic grading may improve the assessments of liver grafts of DBD.
9.Oncological outcomes of laparoscopic radical trachelectomy for early stage cervical cancer
Qing LIU ; Yuxin SUN ; Kaijiang LIU ; Peiquan LI ; Xuanxuan ZHAO ; Zhijun HU
Chinese Journal of Obstetrics and Gynecology 2024;59(2):135-142
Objective:To analyze and summarize the oncological outcomes after laparoscopic radical trachelectomy (LRT) for early stage cervical cancer.Methods:The clinical data and follow-up results of 148 patients with early stage cervical cancer who underwent LRT in Renji Hospital, School of Medicine, Shanghai Jiao Tong University from July 2014 to June 2023 were collected, while tumor outcomes and postoperative pregnancy were analyzed retrospectively.Results:(1) General situation: the median age of 148 patients with LRT was 33 years (range: 19-42 years). Pathological type: 111 cases of squamous cell carcinoma, 36 cases of adenocarcinoma, 1 case of adenosquamous carcinoma. International Federation of Gynecology and Obstetrics (2018) stage: 17 cases of stage Ⅰa1 with lympho-vascular space invasion, 25 cases of stage Ⅰa2, 102 cases of stage Ⅰb1, and 4 cases of stage Ⅰb2. (2) Tumor outcomes: 148 patients were followed up regularly after LRT, and the median follow-up time was 59 months (range: 2-104 months). During the follow-up period, 5 cases of tumor recurred (including 1 death), and the median recurrence time was 10 months (range: 4-33 months). Among them, there were 3 cases of pelvic metastasis, 1 case of distant metastasis, and 1 case of both pelvic and distant metastasis. Both 3-year and 5-year disease-free survival rates of 148 patients were 94.5%, and the 5-year overall survival rate was 98.9%. (3) Postoperative pregnancy: among 148 patients with LRT, 67 patients had pregnancy requirements, followed up for 1 year, and 20 of them were pregnant, with a pregnancy rate of 29.9% (20/67). Among the 20 pregnant patients, 2 cases early abortion, 1 case mid-term abortion, and 17 cases gave birth (including 4 cases of premature birth and 13 cases of full-term birth).Conclusion:Under the condition of strict control of surgical indications, guaranteed surgical scope and tumor-free operation, LRT in patients with early cervical cancer has a good outcome.
10.Construction of and Protein Preparation from a Recombinant Plasmid Containing the Distal Fragment of the Distal C-terminus of the Cav1.2 Channel
Huiyuan HU ; Shuai LEI ; Deri SUN ; Xuanxuan SUN ; Shan YAN ; Shihao LIU ; Jian WANG ; Ying WANG ; Yue LI ; Liying HAO
Journal of China Medical University 2017;46(10):865-868
Objective To construct a recombinant plasmid vector containing the distal fragment of the distal C-terminus (dDCT) of the Cav 1.2 channel,and express,extract,and purify dDCT protein and characterize its biological activity.Methods dDCT cDNA was ligated into the pGEX-6p-1 vector to create a recombinant plasmid that was subsequently transformed into Escherichia coli BL21 competent cells.Expression of GST-dDCT fusion protein from this plasmid was induced with isopropy-β-D-thiogalactoside,and the resulting protein was purified using glutathione-sepharose 4B beads.The biological activity of dDCT was analyzed by GST pull-down assay.Results The recombinant plasmid was verified by restriction enzyme digestion and sequencing.The concentration and purity of the dDCT protein,which was extracted by ultrasonication,were high enough to detect dDCT activity.The binding of dDCT to CT1 was determined to be concentration-dependent.Conclusion The recombinant dDCT plasmid was successfully constructed,providing the fundamental basis for future studies on mechanisms of Cav 1.2 channel autoregulation.