1.Tumor necrosis factor-related apoptosis ligand induces apoptosis in prostate cancer PC-3M cell line.
Zhaohui, CHEN ; Huafang, WANG ; Longjie, GU ; Zhewei, YE ; Yajun, XIAO
Journal of Huazhong University of Science and Technology (Medical Sciences) 2005;25(4):442-4, 447
To study the effect of tumor necrosis factor-related apoptosis inducing ligand (TRAIL) on PC-3M cell line, PC-3M cell line was incubated with gradient concentrations of TRAIL for 4--24 h. Annixin-V fluorescence staining and TUNEL method were employed to detect the apoptosis of PC-3M cells. The morphology of apoptotic PC-3M cells was observed by electron microscopy. The relationship between TRAIL concentrations and the percentage of apoptotic cells was evaluated by flow cytometry. The proliferation inhibitory ratio was calculated by using MTT colorimetry. Our results showed that apoptosis of PC-3M cells could be induced by treatment with TRAIL for at most 4 h. The results of flow cytometry and MTT colorimetry demonstrated a time- and concentration-dependent relationship between cell apoptosis rate and TRAIL concentration. It is concluded that apoptosis of PC-3M cells can be induced by TRAIL. Because of the selective killing effect of TRAIL on tumor cells, it may become a potential alternative for the treatment of advanced prostate cancer.
Apoptosis/*drug effects
;
Cell Line, Tumor
;
Prostatic Neoplasms/metabolism
;
Prostatic Neoplasms/*pathology
;
TNF-Related Apoptosis-Inducing Ligand/*pharmacology
2.The effect of β-elemene combined with irradiation on DNA damage and repair in A549 cells
Longjie LI ; Lijuan ZOU ; Zhuo ZHANG ; Qi WANG
Chinese Journal of Radiation Oncology 2011;20(4):355-357
Objective To study if β-elemene can increase radiation-induced deoxyribonucleic acid (DNA) damage and decrease the damage repair.Methods Exponentially growing human lung adenocarcinoma cells (A549) were exposed to 10 or 20 μg/ml β-elemene for 24 h before irradiation.The effect of β-elemene on the in vitro radiosensitivity of A549 cells was evaluated using clonogenic assay.DNA damage and repair were evaluated using comet assay.Results Exposure to β-elemene before irradiation increased the radiosensitivity of A549 cells.The SERD0 for 10 μg/ml and 20 μg/ml β-elemene was 1.55 and 1.64, respectively.The SERDq for 10 μg/ml and 20 μg/ml β-elemene was 1.43 and 1.75, respectively.Combined treatment, comparing to irradiation or β-elemene treatment alone, induced higher levels of DNA damage and slower rate of damage repair.A549 cells exposed to 20 μg/ml β-elemene followed by irradiation showed a higher levels of tail moment (TM) than those exposed to irradiation or β-elemene alone at 0 h,2 h,6 h and 24 h after irradiation.The TM of the three groups at 0 h,2 h,6 h and 24 h after irradiation was 7.16±2.61,0.95±0.65 and 1.81±1.23(F=231.24,P<0.01), 3.65±2.06,0.11±0.07 and 1.58±1.40(F=90.22,P<0.01), 2.09±0.83,0.1±0.05 and 0.45±0.25(F=238.44,P<0.01), 1.45±1.37,0.11±0.08 and 0.60±0.40(F=38.94,P<0.01), respectively. Conclusions β-elemene can enhance the radiosensitivity of A549 cells through the enhancement of DNA damage and the inhibition of DNA damage repair.
3.Protection against myocardial ischemia-reperfusion injury in rats by a novel selective lymphotoxin binding p55TNFR
Yanhua WANG ; Wenli SU ; Ping HAO ; Wenxian ZHU ; Weiyi SHAO ; Longjie QIAN ; Yixin WANG
Chinese Journal of Emergency Medicine 2015;24(10):1131-1134
Objective To investigate the effect of a novel lymphotoxin with selectively binding to p55 tumor necrosis factor receptor (p55TNFR) on myocardial ischemia-reperfusion injury in rats in order to explore the mechanism.Methods A total of 40 SD rats were randomly (random number) assigned into four groups (n =10 in each),namely sham operation group (group A),I/R group (group B),wild type rhLTα treatment group (group C),and p55TNFR selective rhLTα (rhLTα-Q107E) treatment group (group D).After I/R model rats were established,various therapeutic agents or saline were given by continuous intravenous infusion for 24 h via a micropump.After 24 hours of treatment,serum myocardial zymogram,such as aspartate aminotransferase (AST),lactate dehydrogenase (LDH) and creatine kinase (CK),as well as superoxide dismutase (SOD),malondialdehyde (MDA) and glutathione peroxidase (GSH-Px) activities were determined.Myocardial infarction size (MIS) was measured by nitro blue tetrazolium chloride (NBT) staining.Results Compared to sham operation group,MIS,AST,LDH,CK,MDA were increased,while the activities of SOD and GSH-Px were decreased.However,all the effects were significantly reversed by treatment with rhLTα-Q107E (P < 0.05) but not rhLTα (P > 0.05).Conclusions The rhLTα-Q107E plays a role in the protection against myocardial ischemia-reperfusion injury in rats by the mechanism of scavenging oxygen free radicals and increasing the activity of endogenous antioxidant system.
4.Tumor inhibitory effects of 131I-Trastuzumab on human epidermal growth factor receptor 2 overexpressing breast cancer cells and its possible mechanisms
Longjie ZHANG ; Helei HOU ; Guoming WANG ; Zhenzhen HAN ; Xiaochun ZHANG ; Shengli YUAN
Chinese Journal of Nuclear Medicine and Molecular Imaging 2015;35(4):293-297
Objective To explore antitumor effect of 131I-Trastuzumab on human epidermal growth factor receptor(HER) 2 overexpressing breast cancer cells and investigate its possible mechanism.Methods The expression levels of HER2 of three different breast cancer cell lines (BT474,MCF-7,HCC1937) were detected with immunofluorescence.Trastuzumab was labeled with 131I using the Iodogen method and 131I-Trastuzumab was isolated with ultrafiltration membrane,then the labeling efficiency,radiochemical purity and immunoreactivity were measured.The effects of 131I,Trastuzumab and 131I-Trastuzumab on viability of BT474 cells were evaluated with cell counting kit-8 (CCK-8) assay.The levels of total Akt and phosphorylated Akt (p-Akt) were detected with Western blot analysis.One-way analysis of variance (ANOVA),ANOVA for factorial design,Bonferroni correction and Pearson correlation analysis were used for data analysis.Results The expression level of HER2 in BT474 cells was much higher than those in HCC1937 and MCF-7 cells.The labeling efficiency,radiochemical purity and immunoreactivity of 131I-Trastuzumab were (89.71± 2.93)%,(91.80±1.43)% and (58.84±3.35)% respectively.131I (4.625 GBq/L),Trastuzumab(125.0 rmg/L) and 131I-Trastuzumab(4.625 GBq/L) exhibited a dose-dependent cytotoxicity against BT474 cells (r =-0.964,-0.912,-0.618;all P<0.05).The cell viability of 131I-Trastuzumab treated gourp (34.73% ±5.03%) was significantly lower than those of 131I and Trastuzumab treated groups (64.36%± 1.51% and 58.09%±4.14%;t=10.373 and 8.180,both P<0.05),and the cell viability of control group was (100.00±4.54)%.131I-Trastuzumab shown a positive multiplicative interaction between 131I and Trastuzumab (F=9.226,P<0.05;CDI =0.929).Western blot results showed that there was no significant difference of total Akt expression among the control group,131I group,Trastuzumab group and 131I-Trastuzmab group (F=0.208,P>0.05).P-Akt expression in both Trastuzumab group and 131I-Trastuzumab group were much lower than those of control group and 131I group (t=12.524,15.984,7.347,10.807;all P<0.05),while there was no significant difference of p-Akt expression between Trastuzumab group and 131I-Trastuzumab group(t =3.460,P>0.05).Conclusions 131I-Trastuzumab may kill HER2 overexpressing breast cancer cells more effectively than Trastuzumab alone.The underlying mechanism may be attributed to that 131I-Trastuzumab may enhance the radiosensitivity by the inhibitory effect on PI3K/Akt pathway and thus exert synergistic effects with 131I.
5.Posterior only approaches versus anterior only approaches and combined posterior and anterior approaches for thorac-ic tuberculosis in adults:minimum 5-year follow-up
Hongqi ZHANG ; Longjie WANG ; Mingxing TANG ; Qile GAO ; Jinyang LIU ; Jianhuang WU ; Jianzhong HU
Chinese Journal of Orthopaedics 2016;36(11):641-650
Objective To compare the minimum 5?year follow?up outcomes of surgical management by posterior only ap?proaches, anterior only approaches and combined posterior and anterior approaches for thoracic tuberculosis in adults, and evalu?ate the mid term follow?up results of posterior only approaches. Methods All of 184 patients with monosegment thoracic tubercu?losis between January 2003 and November 2010 were studied retrospectively. Among these patients, 62 cases were treated with posterior debridement combine with interbody fusion (PO group), 65 cases were treated by posterior instrumentation, anterior de?bridement and bone graft in one or two?stage procedures (AP Group ), and 57 cases were treated by anterior only approach (AO Group). The operation time, blood loss, Visual Analogue Scale, complications, recovery of neurological function, kyphosis angle, correction rate and loss angle were respectively compared between each group. Results Comparison of postoperative curative ef?fects showed:mean operation time and blood loss:PO group (260.05±30.75 min,735.95±161.43 ml) was better than AP group (411.65 ± 55.61 min, 1178.65 ± 184.50 ml)and AO group (343.65 ± 24.74 min, 965.35 ± 122.59 ml);corrective angle and correction rate:PO group (6.78°±1.13°, 72.48%±12.97%) and AP group (6.97°±1.05°, 73.10%±11.42%) were better than AO group (13.98°± 1.73°, 44.95%±16.84%);bed time:PO group and AO group were shorter than AP group. Mid term follow?up outcomes showed:ky?phosis angle and loss angle:PO group (8.56°±1.09°, 1.89°±1.41°) and AP group (8.55°±1.65°, 1.63°±1.11°) were better than AO group (16.39°±1.59°, 2.80°±1.29°);bone fusion time, VAS and recovery of neurological function:there were no statistically differ?ence in all groups. Conclusion The mid term follow?up outcomes of posterior debridement combined with interbody fusion is sat?isfied in the management of monosegment thoracic tuberculosis. It is a safe and effective method.
6.Exploration and practice of student' clinical thinking ability in the courses of Medical Laboratory Technology
Yulan LIN ; Bin YANG ; Shoutao CHEN ; Longjie GAN ; Fenqing WANG ; Qiang YI ; Jing CHEN ; Yuefei MA ; Liqin GAO ; Qishui OU
Chinese Journal of Medical Education Research 2017;16(4):381-384
This study was conducted to explore a proper training model of interns' clinical thinking ability under the construction of a new four-year system of medical laboratory technology courses, combined with the establishment of innovative standard whole process practice mode. Multi-teaching methods of clinical thinking, such as explanation of laboratory sheet, interactive teaching based on micro digital system, interdisciplinary multiple information system, combined PBL teaching and intern report, were applied and evaluated in the laboratory. Integrated application of these methods remarkably improved the intern's com-prehensive professional quality and their practice performance. All methods received high evaluation from both the interns and teachers.
7.Tumor Necrosis Factor-related Apoptosis Ligand Induces Apoptosis in Prostate Cancer PC-3M Cell Line
Zhohui CHEN ; Huafang WANG ; Longjie GU ; Zhewei YE ; Yajun XIAO
Journal of Huazhong University of Science and Technology (Medical Sciences) 2005;25(4):442-444,447
To study the effect of tumor necrosis factor-related apoptosis inducing ligand (TRAIL)on PC-3M cell line, PC-3M cell line was incubated with gradient concentrations of TRAIL for 4-24h. Annixin-Ⅴ fluorescence staining and TUNEL method were employed to detect the apoptosis of PC-3M cells. The morphology of apoptotic PC-3M cells was observed by electron microscopy. The relationship between TRAIL concentrations and the percentage of apoptotic cells was evaluated by flow cytometry. The proliferation inhibitory ratio was calculated by using MTT colorimetry. Our results showed that apoptosis of PC-3M cells could be induced by treatment with TRAIL for at most 4 h. The results of flow cytometry and MTT colorimetry demonstrated a time- and concentration-dependent relationship between cell apoptosis rate and TRAIL concentration. It is concluded that apoptosis of PC-3M cells can be induced by TRAIL. Because of the selective killing effect of TRAIL on tumor ceils, it may become a potential alternative for the treatment of advanced prostate cancer.
8.Predictive value of vertebral trabecular and endplate Hounsfield Units on cage subsidence followed posterior lumbar interbody fusion
Hui WANG ; Da ZOU ; Zhuoran SUN ; Longjie WANG ; Shuai JIANG ; Weishi LI
Chinese Journal of Orthopaedics 2021;41(13):864-871
Objective:To explore the predictive value of vertebral trabecular and endplate HU values on cage subsidence after posterior lumbar interbody fusion (PLIF), hope to provide reference for surgical planning.Methods:All of 72 patients with lumbar disc herniation that underwent PLIF were retrospectively reviewed, who were divided into two groups according to the occurrence of cage subsidence at one-year follow up. Cage subsidence was defined as more than 4 mm subsidence into the vertebrae valuated by CT at one-year follow up. There were 18 patients enrolled into Subsidence group and 54 patients enrolled into N-Subsidence group. The lumbar lordosis, segmental lordosis, intervertebral height, off-bed time, hospital stay, complications, the trabecular and endplate HU values of upper instrumented vertebrae (UIV) and lower instrumented vertebrae (LIV) were compared between the two groups. ROC was used to explore the thresholds of HU values.Results:There were 14 patients presented cage subsidence into the L4, 4 patients presented cage subsidence into the L5. There was no significant difference in lumbar lordosis, segmental lordosis, intervertebral height, off-bed time, hospital stay, or complications between the two groups. Both UIV and LIV trabecular and endplate showed a lower HU value in Subsidence group than those in N-Subsidence group. The most appropriate thresholds of HU value were 146, 172, 307, 254 for trabecular of UIV, trabecular of LIV, lower endplate of UIV, and upper endplate of LIV, respectively.Conclusion:Vertebral trabecular and endplate HU values could effectively predict the cage subsidence after PLIF, patients should be completely informed the risk of cage subsidence and larger cage should be recommended if they presented HU values under the certain threshold.
9.SRT1720,an activator of silent information regulator 1,alleviates acute traumatic brain injury in a rat model
Longjie QIAN ; Wenli SU ; Wenxian ZHU ; Yixin WANG
Chinese Journal of Tissue Engineering Research 2024;28(28):4447-4454
BACKGROUND:It has been shown that in a mouse model of acute traumatic brain injury,the transcriptional and translational levels of silent information regulator 1(SIRT1)activated by drugs significantly elevates the expression of SIRT1 in brain tissue,reduces inflammatory and oxidative stress in brain tissue,and improves neurological function. OBJECTIVE:To investigate the mechanism of intraperitoneal injection of SRT1720,an activator of SIRT1,to alleviate acute traumatic brain injury in rats. METHODS:Ninety Sprague-Dawley rats were randomized into three groups(n=30 per group):a sham group(without modeling),a model group and an activator group.Animal models of acute traumatic brain injury were established in the latter two groups.At 6 hours after modeling,the sham,model and activator groups were injected intraperitoneally with dimethyl sulfoxide solution,methylsulfoxide solution and SRT1720 once a day for 28 days,respectively.The time points for sampling were set,and rats'neurological function,brain tissue water content,brain tissue oxidative stress and inflammatory response,brain tissue morphology,apoptosis and angiogenesis,and the protein expression of SIRT1 in brain tissue were detected and measured. RESULTS AND CONCLUSION:Compared with the sham group,the modified neurological deficit score,brain tissue water content and apoptosis rate of rats were increased in the model group at 7,14 and 28 days of injection(P<0.05);compared with the model group,the modified neurological deficit score,brain tissue water content and apoptosis rate of rats were decreased in the activator group(P<0.05).Compared with the sham group,the levels of reactive oxygen radicals and myeloperoxidase in the brain tissue were increased(P<0.05),the levels of malondialdehyde,tumor necrosis factor α and interleukin 6 in the serum were increased(P<0.05),and the levels of superoxide dismutase in the serum were decreased in the model group at 7,14 and 28 days of injection(P<0.05).Compared with the model group,the levels of reactive oxygen radicals and myeloperoxidase in the brain tissue were decreased(P<0.05),the levels of malondialdehyde,tumor necrosis factor α and interleukin 6 in the serum were decreased(P<0.05),and the levels of superoxide dismutase in the serum were increased in the activator group at 7,14 and 28 days of injection(P<0.05).Immunohistochemical staining at 7,14 and 28 days of injection showed that the number of new vessels in the brain tissue was higher in the model group than the sham group(P<0.05)as well as higher in the activator group than the model group(P<0.05).Western blot assay indicated that at 7,14 and 28 days of injection,the expression of SIRT1 protein in the brain tissue was lower in the model group than the sham group(P<0.05)and higher in the activator group than the model group(P<0.05).Hematoxylin-eosin staining showed that at 7,14 and 28 days of injection,the degree of brain injury in the activator group was less than that in the model group.To conclude,intraperitoneal injection of the SIRT1 signal activator SRT1720 can significantly reduce oxidative and inflammatory stress in the brain tissue,inhibit neuronal apoptosis,promote angiogenesis,and alleviate brain injury in rats with acute traumatic brain injury.