1.Cisplatin induces oxidative stress and apoptosis in pericytes of cochlear stria vascularis via mitochondria-mediated pathway
Tianlan HUANG ; Rongkui CHAI ; Tianfeng SHI ; Jingwen MA ; Meng YU ; Miao YU ; Junqiang SI ; Li LI
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2023;58(11):1093-1101
Objective:To study whether cisplatin may induce apoptosis in the pericytes of cochlear stria vascularis and underlying mechanisms.Methods:Twenty male C57BL/6J mice aged 6-8 weeks were divided into control group and a cisplatin group. Primary cultured mouse cochlear vascular peristriatal cells were identified and divided into control group, cisplatin group, and N-acetylcysteine+cisplatin group. Auditory brainstem response (ABR) was used to detect hearing in mice. Hematoxylin eosin (HE) staining was used to observe morphological changes in the stria vascularis of the cochlea. The total superoxide dismutase (SOD) activity test kit (WST-1 method) and thiobarbituric acid (TBA) method were used to detect SOD activity and Malondialdehyde (MDA) content, respectively. DCFH-DA fluorescence probe was used to detect the content of reactive oxygen species in peripheral cells. Hoechst 33 342 and flow cytometry were used to detect the apoptosis rate of pericytes. Immunofluorescence technology was used to detect the distribution and expression of apoptosis related proteins in pericytes of cochlear stria vascularis. Immunohistochemistry and Western blotting (WB) were used to detect the expression of apoptosis-and mitochondrial-related proteins. Mito SOX TM-red and JC-1 were used to detect the mitochondrial function of pericytes. Evans blue staining was used to observe the permeability of the blood labyrinth barrier (BLB). Statistical analysis was conducted using SPSS 18.0 software. Results:Compared with the control group, the cisplatin group significantly increased in the hearing threshold ( t=4.72, P<0.01), Ⅰ-wave latency ( t=12.25, P<0.05), and the levels of oxidative stress in the cochlea and pericytes ( t=38.34, P<0.01), and also cisplatin caused disorder and contraction of the cochlear stria vascularis structure, increased BLB permeability [Evans blue leakage (1.08±0.42) AU vs (0.55±0.23) AU, t=4.64, P<0.05], with a statistically significant difference, enhanced the expressions of apoptotic proteins c-Caspase-3 ( t=5.01, P<0.01) and Bax ( t=6.33, P<0.01) in the peristriatal cells of cochlear blood vessels in mice treated with cisplatin increased. And cisplatin can induce apoptosis of primary cultured pericytes and up-regulate the expression of c-Caspase-3 and Bax ( P<0.05). The NAC+cisplatin group partially reversed cisplatin-induced pericyte apoptosis ( P<0.05). Cisplatin caused damage to the mitochondrial function of peripheral cells, and induced the release of apoptosis-inducing factor (AIF) and cytochrome C (cyt-c) into the cytoplasm ( P<0.05). The NAC+cisplatin group partially reversed cisplatin induced pericyte apoptosis ( P<0.05) and mitochondrial damage ( P<0.05). Conclusion:Cisplatin can increase the level of oxidative stress in the cochlea and cause mitochondrial pathway apoptosis in C57BL/6J mouse cochlear vascular peristriatal cells.
2.Comparison of segmentectomy versus lobectomy for ≤2 cm lung adenocarcinoma with micropapillary and solid subtype negative by intraoperative frozen sections: A multi-center randomized controlled trial
Chang CHEN ; Yuming ZHU ; Gening JIANG ; Haifeng WANG ; Dong XIE ; Hang SU ; Long XU ; Deping ZHAO ; Liang DUAN ; Boxiong XIE ; Chunyan WU ; Likun HOU ; Huikang XIE ; Junqiang FAN ; Xuedong ZHANG ; Weirong SHI ; Honggang KE ; Lei ZHANG ; Hao WANG ; Xuefei HU ; Qiankun CHEN ; Lei JIANG ; Wenxin HE ; Yiming ZHOU ; Xiong QIN ; Xiaogang ZHAO ; Hongcheng LIU ; Peng ZHANG ; Yang YANG ; Ming LIU ; Hui ZHENG
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2021;28(11):1292-1298
Objective To compare the clinical effects of segmentectomy and lobectomy for ≤2 cm lung adenocarcinoma with micropapillary and solid subtype negative by intraoperative frozen sections. Methods The patients with adenocarcinoma who received segmentectomy or lobectomy in multicenter from June 2020 to March 2021 were included. They were divided into two groups according to a random number table, including a segmentectomy group (n=119, 44 males and 75 females with an average age of 56.6±8.9 years) and a lobectomy group (n=115, 43 males and 72 females with an average of 56.2±9.5 years). The clinical data of the patients were analyzed. Results There was no significant difference in the baseline data between the two groups (P>0.05). No perioperative death was found. There was no statistical difference in the operation time (111.2±30.0 min vs. 107.3±34.3 min), blood loss (54.2±83.5 mL vs. 40.0±16.4 mL), drainage duration (2.8±0.6 d vs. 2.6±0.6 d), hospital stay time (3.9±2.3 d vs. 3.7±1.1 d) or pathology staging (P>0.05) between the two groups. The postoperative pulmonary function analysis revealed that the mean decreased values of forced vital capacity and forced expiratory volume in one second percent predicted in the segmentectomy group were significantly better than those in the lobectomy group (0.2±0.3 L vs. 0.4±0.3 L, P=0.005; 0.3%±8.1% vs. 2.9%±7.4%, P=0.041). Conclusion Segmentectomy is effective in protecting lungs function, which is expected to improve life quality of patients.
3.Study on NAEK-labelled HiD-Hin47 fusion protein as a carrier for pneumococcal polysaccharide conjugate vaccine
Hanyang GAO ; Hechu LIANG ; Lin XU ; Haomeng WANG ; Zhihong YAN ; Jianming SHI ; Junqiang LI ; Tao ZHU
Chinese Journal of Microbiology and Immunology 2021;41(11):880-886
Objective:To develop an unnatural amino acid-labelled HiD-Hin47 fusion protein as a novel carrier.Methods:Twenty versions of the fusion protein were designed, each of which contained a different amino acid site replaced by an azide-bearing amino acid, N6-(2-azidoethoxy) (carbonyl)-L-Lysine (NAEK). These fusion proteins were constructed, expressed and purified, and the yield were evaluated by SDS-PAGE. Based on the highest protein yield, which was approximately 70% of the wild-type yield, the fusion protein with the unnatural amino acid in E677 site was selected. The pneumococcal polysaccharides of serotype 3 (F3) and serotype 6B (F6B) were coupled to the selected fusion protein through a "click" reaction. The conjugates were purified and compared in animal studies with other F3 and F6B conjugates that were coupled to CRM197, tetanus toxoid (TT) and HiD by conventional methods.Results:The immunogenicity of F3 conjugate using HiD-Hin47 as carrier (F3-HiD-Hin47) was slightly better than that of other F3 conjugates. F6B-HiD-Hin47 was significantly better than F6B-TT and F6B-HiD in terms of immunogenicity, but showed no significant difference with F6B-CRM197.Conclusions:NAEK-labelled HiD-Hin47 had the potential as a carrier for pneumococcal polysaccharide conjugate vaccine and was worthy of further study.
4.Effects of cell proliferation and apoptosis of L-carnitine combined with epirubicin on different human lung adenocarcinoma cells
Yintao YE ; Wei ZHONG ; Junqiang QIAN ; Yun SHI ; Chen WANG
International Journal of Biomedical Engineering 2019;42(1):15-20
Objective To investigate the effect of L-carnitine (LCNT) combined with epirubicin (EPI) on cell proliferation and apoptosis of two lung adenocarcinoma cell lines (GLC-82 and A549). Methods GLC-82 and A549 cells were divided into control group, EPI group, LCNT group and EPI+LCNT group, respectively. The cell proliferation rate was examined by MTT assay 24 h after the treatment, and the cell cycle and cell early apoptosis were analyzed by flow cytometry. The expression of P53 and Bcl-2 proteins were detected by Western Blot. Results Compared with the control group, the proliferation rate of GLC-82 and A549 cells was 37.56%(P<0.01) and 6.32%(P>0.05) after 24 hours of 20.00μg/ml LCNT treatment indicating LCNT could promote the proliferation of GLC-82 cells. Compared with the EPI group, EPI+LCNT had smaller inhibitory effect on the proliferation of GLC-82 cells, and the inhibition rate of the EPI+LCNT group was 12.14%lower than that of EPI group (P<0.05). Compared with the EPI group, EPI+LCNT could significantly increase the G0/G1 phase ratio of GLC-82 cells (50.42%±1.21%vs. 25.94%± 0.66%, P<0.01) and A549 cells (54.92%±1.71%vs. 38.63%±0.69%, P<0.01), decrease the S phase ratio of GLC-82 cells (34.21%±0.96%vs. 59.68%±1.25%, P<0.05), and prevent the early apoptosis of GLC-82 cells without affecting apoptosis of A549 cells. Moreover, in the EPI+LCNT group, the expression of P53 protein in GLC-82 and A549 cells was down-regulated (P<0.05), and the expression of Bcl-2 protein in GLC-82 cells was up-regulated (P<0.01) and no significant change in A549 cells (P>0.05). Conclusions Comparing with the EPI, the combination of LCNT and EPI has less proliferation inhibition and apoptosis induction on GLC-82 cells, and without significant effect on A549 cells. LCNT can promote the proliferation of GLC-82 cells and block the cell cycle at G0/G1 phase. This mechanism may be related to down-regulation of P53 protein and up-regulation of Bcl-2 protein expression.
5.Time of lower extremity deep venous thrombosis after hip arthroplasty in senile patients with osteoporotic femoral neck fractures
Hao SUN ; Junqiang WEI ; Lirui LIU ; Shi YAN ; Yu JIN ; Zhen FENG
Chinese Journal of Tissue Engineering Research 2017;21(19):2961-2965
BACKGROUND: Lower extremity deep venous thrombosis is commonly seen after hip arthroplasty. The time of thrombosis after arthroplasty in senile patients with osteoporotic femoral neck fractures is little reported.OBJECTIVE: To investigate the time of perioperative lower extremity deep venous thrombosis in senile patients with osteoporotic femoral neck fractures undergoing hip arthroplasty. METHODS: The data of 178 patients with femoral neck fractures undergoing hip arthroplasty were analyzed, and were then allocated to anticoagulant and non-anticoagulant groups according to the preoperative use of low-molecular-weight heparins or not. Doppler ultrasonography of both lower extremity arteries was conducted at 1, 3, 5 and 5-7 days after injury, and 2, 5, 7 and 14 days postoperatively to monitor the lower extremity deep venous thrombosis, and the number and incidence of thrombosis in the two groups were recorded at different time points. The time of thrombosis occurred frequently after injury and surgery was determined through drawing the frequency distribution table and line chart.RESULTS AND CONCLUSION: (1) The incidence of thrombosis showed significant difference between anticoagulant (n=6, 8%) and non-anticoagulant (n=22, 21%)) groups (P < 0.05). (2) Thrombosis frequently occurred at 3 days after injury and 1 day postoperatively. (3) These results suggest that the thrombosis in patients with femoral neck fractures frequently occurs at 1-3 days after injury and 1 day after replacement. Additionally, active prevention measures after injury can significantly reduce the incidence of lower extremity venous thrombosis.
6.Effect of transcription factor SOX18 on proliferation, migration and invasion of cervical cancer cell line HeLa
Junqiang SHI ; Jianli WANG ; Guofang WANG ; Chunmei WANG
Cancer Research and Clinic 2017;29(6):370-373
Objective To explore the effect of transcription factor SOX18 on proliferation, migration and invasion of HeLa cells and its related mechanism. Methods HeLa cells were divided into blank control group, negative control group, pCI-SOX18 group. The expression of SOX18 and matrix metalloproteinase 7 (MMP-7) was detected by Western blot. The cell proliferation of different group was measured by MTT analysis. The effect of transcription factor SOX18 on migration and invasion of HeLa cells were measured by Transwell assay. Results HeLa cells were successfully transfected with pCI-SOX18. According to the results of MTT analysis, the cell number of pCI-SOX18 group showed no significant difference compared with blank control group and negative control group (P> 0.05). Compared to blank control group (0.23 ±0.05) and negative control group (0.28±0.07), SOX18 expression in pCI-SOX18 group (1.16±0.13) was significantly higher (F=218.14, P<0.05). Meanwhile, MMP-7 expression in pCI-SOX18 group (0.95±0.09) was increased significantly compared with blank control group (0.71±0.07) and negative control group (0.68±0.06) (F=116.84, P < 0.05). The number of cell migration in pCI-SOX18 group (175.71 ±17.17) was increased significantly compared with that in negative control group (95.19 ±8.18) and that in blank control group (93.34±9.72) (t=75.47, t=77.35, P<0.05). Compared with that in negative control group (98.63±14.62) and that in blank control group (96.57 ±13.95), the number of cell invasion in pCI-SOX18 group (247.12 ±36.82) was significantly higher (t=98.02, t=99.34, P<0.05). Conclusion Overexpression of SOX18 has no effect on the proliferation of HeLa cells, but has a significant role in promoting cell migration and invasion probably by regulating MMP-7 expression positively.
7.Proximal femoral nail antirotation fixation for osteoporotic intertrochanteric fracture in the elderly:characteristics of deep venous thrombosis of lower extremity
Junqiang WEI ; Lirui LIU ; Xinyu WANG ; Shi YAN ; Yu JIN ; Zhen FENG
Chinese Journal of Tissue Engineering Research 2016;20(35):5224-5230
BACKGROUND:The risk of lower-extremity deep venous thrombosis was high in patients with the osteoporotic hip fracture. The aged age and hip fracture surgery are major risk factors for thrombosis. No study concerned the critical point in time course to the lower extremity deep venous thrombosis in the osteoporotic hip fracture patients.
OBJECTIVE:To analyze the incidence of the perioperative deep venous thrombosis of lower extremity limb at different time slots in senile osteoporotic intertrochanteric fracture patients repaired by proximal femoral nail antirotation fixation.
METHODS:From June 2012 to January 2016, 220 cases diagnosed as osteoporotic intertrochanteric fracture and performed closed reduction and proximal femoral nail antirotation fixation were selected from the Department of Orthopedics, the Affiliated Hospital of Chengde Medical Col ege in China. Based on administered low-molecular-weight heparins (LMWH) or not after trauma, there were two groups:LMWH group and the non-LMWH group. Vascular Doppler ultrasound was conducted at 1, 3 and 5 days after trauma, more than 5 days after trauma, 2, 5, 7 and 14 days after surgery. The total number of cases and incidence of thrombosis were recorded and compared between the two groups. The number of cases and the incidence of new thrombosis were recorded at different time points in both groups. Frequency distribution table was used to analyze the time of suffering thrombosis after injury and surgery.
RESULTS AND CONCLUSION:(1) Among 220 cases, 18 cases in the LMWH group (n=154) affected thrombosis, with an incidence of 12%, and 15 cases in the non-LMWH group (n=66) experienced deep vein thrombosis, with an incidence of 23%. Significant differences in the incidence of thrombosis were detected between the two groups (P=0.041). (2) Peak time of thrombus was 3 days before injury and 1 day after surgery in both groups. (3) Results suggested that the incidence of deep vein thrombosis was high in proximal femoral nail antirotation fixation for intertrochanteric fracture. The peak time of deep vein thrombosis was 3 days before injury and 1 day after surgery. The prevention of deep vein thrombosis should begin from injury. Prophylactic anticoagulation after injury can dramatical y decrease the incidence of deep vein thrombosis.
8.Relationship of quadriceps tendon and the stability of external fixator in rabbit femur fracture model
Meng ZHANG ; Junqiang WEI ; Jianwei DUAN ; Shi YAN
Chinese Journal of Tissue Engineering Research 2016;20(39):5834-5839
BACKGROUND:In rabbit femur fracture model experiment, there are a few studies on whether quadriceps tendon wil affect the stability of the fixed device. OBJECTIVE:To analyze the effects of quadriceps tendon to the stability of external fixator in rabbit models of femur fracture. METHODS:The 48 New Zealand white rabbits were randomly divided into two groups. External fixator was used to make fracture model. In the experiment group, quadriceps tendon was cut. In the control group, quadriceps tendon was retained. 2 weeks later, X-ray examination was utilized to observe the number of failed external fixator in both groups from modeling to success. RESULTS AND CONCLUSION:(1) X-ray examination:2 weeks after model establishment, external fixation frame screw was free from femur in one rabbit of the experiment group, and seven rabbits in the control group. Of them, one screw was broken. Threaded portion retained in the femur. (2) External fixation failure rate:failed external fixator was found in one rabbit in the experiment group, resulting in a failure rate of 4%, and 12 rabbits in the control group, resulting in a failure rate of 50%(P<0.05). These findings suggested that cutting off the rabbit quadriceps tendon can effectively ensure the stability of the external fixator, and indirectly improve success rate of the experiment.
9.MR Tracer Diffusion in Cerebral Interstitial Fluid of Elderly SD Rats
Shuaiwen WANG ; Chunyan SHI ; Long ZUO ; Hongbin HAN ; Junqiang LEI
Chinese Journal of Medical Imaging 2015;(6):409-412
Purpose With the progression of brain tissue aging, the transport and drainage characteristics of metabolites and secretory products for neurons in extracellular space occurs irreversible change. This paper aims to investigate and quantify MR tracer diffusion characteristics in cerebral interstitial fluid of elderly SD rats. Materials and Methods MR contrast agent Gd-DTPA was injected into the caudate nucleus of two groups of rats including 8 in experimental group (15-17 month old) and 15 in control group (7-10 month old). MR scan was performed at 0.25 h, 0.5 h, 1 h, 2 h, 3 h and 4 h to observe the dynamic distribution in the caudate and measure the diffusion and clearance rate. Results There was no statistically significant difference in diffusion rate and D* between control group with (3.32±0.70)×10-4 mm2/s and experimental group with (3.25±0.46)×10-4 mm2/s (t=1.739, P>0.05). The clearance rate k' was significantly different between control group (0.62±0.12)×10-4/s and experimental group (0.29±0.08)×10-4/s (t=11.602, P<0.05). Conclusion The degeneration of aging brain tissue changes the composition of extracellular space resulting in decreased speed of ISF clearance. This may cause accumulation of metabolites which eventually triggers a variety of age-related diseases.
10.Relationship between the Cx37 polymorphism of connexin gene and essential hypertension in Kazak and Han population in Xinjiang
Bihan LU ; Wenwen ZHANG ; Wenyan SHI ; Ketao MA ; Li LI ; Shuxia GUO ; Junqiang SI
Tianjin Medical Journal 2015;(11):1326-1329,1330
Objective To investigate the association of the Cx37 polymorphism of connexin gene with essential hyper?tension (EHT) in Xinjiang Han and Kazak population. Methods In Xinjiang region, 500 EHT patients (EHT group) were in?cluded in this study including Kazak 250 cases and Han 250 cases. Five hundred healthy volunteers (NT) were used as NT group including Kazak 250 cases and Han 250 cases. The values of age, body mass index (BMI), systolic blood pressure (SBP) and diastolic blood pressure (DBP), and other general clinical features were compared between two groups. The poly?morphism of Cx37 gene rs1630310, rs697372 and rs705193 SNP were compared between EHT and NT groups in the two eth?nic groups. Hardy-Weinberg equilibrium was used to detect the representation, and differences of genotype frequencies and gene frequency were calculated in two groups of Kazak and Han groups. Results There were significant differences in BMI, SBP, DBP, apolipoprotein ratios and homocysteine between EHT group and NT group in Kazak and Han groups (P<0.05). There were no significant differences in genotype frequencies and gene frequencies of rs705193 between EHT and NT groups (P>0.05). The differences of Kazak rs1630310 genotype and gene frequency were statistically significant (P<0.01). The frequency of Kazak rs697372 locus genotype was not significantly different (P>0.05), but the difference of gene frequen?cy was statistically significant (P<0.05). There were no significant differences in rs1630310 and rs697372 locus genotype and gene frequency in two groups of Han group. Conclusion Cx37 gene polymorphism is associated with the occurrence of EHT in Xinjiang Kazak population, which may be related with the rs1630310 and rs697372 polymorphism.


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