1.Theory of combinant expression vector of survivin antisense RNA and HSPT0 for the therapy of bladder cancer
International Journal of Surgery 2012;39(5):349-353
Carcinoma of urinary bladder is one of the most common tumor.As life rhythm speeding up and lifestyle changing,bladder cancer incident increases year after year.Previous major treatment methods for the disease are open surgery and TURBT,but these therapies can not overcome the characteristics of its recurrence.Since the 21st century along with the rapid development of molecular biology technology,the discovery of Survivin gene,new knowledge about HSP molecules in tumor immunology and the progress of the technique called RNA interference,open up a whole new field for early diagnosis and treatment of bladder carcinoma.Discussing the role of Survivin gene in the development of tumor,survivin antisense RNA' s efficacy and the role of HSP70 in the cancer immunity,the review aims at exploring the possibility of the double gene expression vector constructed by Survivin antisense RNA gene and HSP70 gene for bladder carcinoma therapy.
2.Establishment and biological characterization of a human glucocorticoid-resistant cell line of diffuse large B cell lymphoma
Wei TAN ; Bobin CHEN ; Yan MA ; Xiaoping XU ; Guowei LIN
Journal of Leukemia & Lymphoma 2013;22(12):712-716
Objective To establish a novel glucocorticoid (GC)-resistant human diffuse large B lymphoma(DLBCL) cell line Toledo/dexamethasone (DEX) by the exposure to DEX,and observe the biological characteristics of resistant and parental cell line,investigate the mechanisms of glucocorticoid-resistance.Methods Toledo/DEX was established by the exposure to DEX,the dose of which was increased gradually and intermittently for long periods of time.Toledo,in the logarithmic growth phage,was incubated in the culture medium containing DEX at the concentration of 1×10-8 mol/L at first.The medium without DEX was replaced after for 96 hours until the cell line re-entered the logarithmic growth phase.Repeat the above steps to acquire the ultimate concentration of DEX in the medium as 1.024 ×10-5 mol/L.The biological characteristics of resistant and parental cell lines were evaluated.Results Toledo/DEX was more invasive in the aspects of ultrastructure,tumorigenicity and drug sensitivity.Meanwhile,Toledo/DEX achieved some stable biological characteristics such as morphology,karyotypes and immunophenotype.Furthermore,GC receptor (GR) α and GR β protein expression analysis showed that GR was involved in the mechanism of the GCresistance.Conclusions Toledo/DEX is a drug-resistant cell line with a stable biology backgroud.These results may help shed light on the knowledge of GC-resistance and lay the groundwork for searching new therapeutics to reverse drug-resistance.
3.Expression of Human Connexin26 in COS-7 Cells
Cong XU ; Guowei LI ; Zeyu LIU ; Jiawei LIAO ; Panpan XUE ; Binyan TAN ; Jiahai LU
Journal of Tropical Medicine 2007;7(5):401-407
Objective To construct the eukaryotic expression plasmid for the expression of human Connexin26 in COS-7 cells.Methods Total RNA was isolated from human peripheral blood lymphocytes and used as template for the PCR cloning of the human Connexin26 gene.The human Cx26 cDNA containing the 678 bp whole coding region of the human Connexin26 gene was amplified by PCR using specific primers and cloned into the pCI-neo vector to construct the recombinant eukaryotic expression plasmid,pCI-Cx26.The recombinant plasmid was identified by restriction endonuclease digestion,and transfected into COS-7 cells by liposome.The expression of Cx26 mRNA and the protein were analyzed by RT-PCR and SDS-PAGE,respectively.Results Restriction endonuclease digestion analysis verified successful construction of the recombinant plasmid,pCI-Cx26.The expression of Cx26 mRNA and protein in the transfected COS-7 cells were detected by RT-PCR and SDS-PAGE,respectively.Conclusion The eukaryotic expression plasmid for human Cx26 has been constructed successfully with the capability of expression in COS-7 cells.
4.Acute stress disorder and related factors in a troop after participating in disaster relief in Beichuan county after Wenchuan earthquake
Guangtao HU ; Xuecheng LI ; Guowei WANG ; Ying HE ; Lan YANG ; Xiaoqiong TAN
Journal of Third Military Medical University 2003;0(15):-
Objective To investigate the occurrence circumstance of acute stress disorder(ASD),clinical features and related risk factors,in the officers and soldiers who had gone to Beichuan county to provide disaster relief.Methods We used the self-general questionnaire,post-traumatic stress symptoms self-rating scale(PCL-C),perceived social support scale(PSSS),and coping style questionnaires(SCSQ)to assess the 126 subjects,combining with semi-structured interviews to determine and compare the ASD group and control group,and then analyzed the related factors.Results The prevalence rate of ASD officers and soldiers was 13.49%,with main symptoms of repeatedly breaking into painful memories,nightmares,reproducing the scenes of traumatic events,strong trouble of psychological pain,decreased interest,sleep disorders and emotional instability.These main symptoms of ASD interacts each other,and were reduced with positive response and good social support.Conclusion In the phase of acute stress(one month),the officers and soldiers taking part in earthquake relief have symptoms of ASD,so we should strengthen the necessary psychological assessment and comprehensive psychological intervention means,in order to slow down psychological trauma after disaster,prevent and reduce the possibility that ASD turns into post-traumatic stress disorder(PTSD).
5.Relationship of bone mineral density and bone metabolism indicators with osteoporosis in elderly patients with post-stroke hemiplegia
Wanying FU ; Yongwen ZHAI ; Guowei TAN ; Hualong YANG ; Hong GU
Chinese Journal of Geriatric Heart Brain and Vessel Diseases 2023;25(12):1306-1309
Objective To explore the relationship of changes in bone mineral density and bone me-tabolism indicators with osteoporosis in elderly patients with hemiplegia after cerebral infarction.Methods A total of 197 elderly patients suffering post-stroke hemiplegia admitted to our hospital were enrolled between February 2018 and February 2021.Their bone mineral density and bone metabolism indicators were analyzed based on gender,disease course and hemiplegia severity.According to disease severity,they were divided into mild group(112 cases),moderate group(62 cases)and severe group(32 cases).Results Among the 197 elderly patients,83(42.13%)had osteoporosis.The males had significantly higher bone mineral density of proximal femur and levels of bone alkaline phosphatase(BALP)and osteocalcin,and lower levels of serum type Ⅰ collagen(CICP)and C-terminal telopeptides of type Ⅰ collagen(CTXI)than the females(P<0.01,P<0.05).Compared with the patients with hemiplegia course for 3-6 months,bone mineral density values of greater trochanter and total hip,and levels of BALP and osteocalcin were obviously de-creased,while levels of CICP and CTXI were notably increased in those with the course of 7-12 months and 13-24 months(P<0.05).Compared with the mild group,bone mineral density val-ues of greater trochanter and total hip,and levels of BALP and osteocalcin were significantly de-creased,while levels of CICP and CTXI were significantly increased in moderate group and severe group(P<0.05).Compared with the moderate group,bone mineral density values of greater tro-chanter,intertrochanteric site and total hip were significantly decreased in the severe group(P<0.05).The bone mineral density of above parts was negatively correlated with CICP levels(r=-0.458,r=-0.403,r=-0.355,r=-0.482,P<0.01)and CTXI levels(r=-0.305,r=-0.291,r=-0.273,r=-0.384,P<0.05,P<0.01),while positively correlated with BGP level(r=0.476,r=0.428,r=0.386,r=0.518,P<0.01).Conclusion The incidence of osteoporosis is quite higher in the elderly patients with post-stroke hemiplegia.There exist bone mineral density decline and abnormal bone metabolism,and their changes are related to gender,course and severi-ty of hemiplegia.
6.Prediction for target genes affecting biological behaviors of glioblastoma
Shuo WANG ; Zheng CHEN ; Yuanyuan XIE ; Guowei TAN
Chinese Journal of Neuromedicine 2022;21(12):1203-1208
Objective:To explore the differentially expressed genes in glioblastoma development and their functions and roles as to identify the target genes influencing biological behaviors of glioblastoma.Methods:Original gene expression profiles of GSE70231 dataset obtained from Gene Expression Omnibus database were screened for differentially expressed genes by GEO2R software. DAVID database was used to conduct gene ontology (GO) function analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis for these differentially expressed genes. STRING database was used to construct protein-protein interaction (PPI) network of these differentially expressed genes; target genes were selected from PPI network by cytoHubba and MCODE plug-ins; GEPIA online was used to analyze the expressions of target genes in glioblastoma and their influences in overall survival of glioblastoma patients (sample data collecting from The Cancer Genome Atlas database). Finally, the selected target genes were verified using RNA-seq dataset GSE50021 from human tissues.Results:Totally, 520 differentially expressed genes were identified, including 305 up-regulated genes and 215 down-regulated ones. GO analysis showed that the differentially expressed genes were mainly enriched in biological processes (signal transduction, cell adhesion, and positive regulation of cell proliferation), cytological components (extracellular exosomes, cytoplasm, and cytoplasmic membrane), and molecular function (protein binding). KEGG pathway enrichment analysis showed that the differentially expressed genes were mainly enriched in mitogen activated protein kinase signal pathway, proteoglycans in cancer, oxytocin signal pathway and calcium signal pathway. Totally, 17 target genes were selected by MCODE and cytoHubba plug-ins from the PPI network of differentially expressed genes; functional analysis and clinical sample verification showed 8 target genes ( VCAM1, SPP1, ITGB1, CTGF, VIM, ITGAV, COL1A1, and BCL2A1) could affect the biological behaviors of glioblastoma; the correlations of ITGAV, COL1A1, and BCL2A1 with glioma had been rarely reported, and GSE50021 dataset verified that their expressions in glioblastoma tissues were significantly higher than those in normal brain tissues ( P<0.05). Conclusion:These 8 target genes concluded from this research, especially BCL2A1, COL1A1 and ITGAV, may be important targets for exploring the pathogenesis, diagnosis and treatment of glioblastoma in the future.
7.Is Laparoscopic Sleeve Gastrectomy for Asian Super Obese a Safe and Effective Procedure?
Hannah Jh NG ; Guowei KIM ; Claire Alexandra Z CHEW ; Moe Thu SAN ; Jimmy By SO ; Asim SHABBIR
Annals of the Academy of Medicine, Singapore 2018;47(5):177-184
INTRODUCTIONOutcomes of bariatric surgery for super obese Asians are not well reported. We aimed to compare short-term outcomes of laparoscopic sleeve gastrectomy (LSG) in Asian patients with body mass index (BMI) <47.5 kg/m to those with BMI ≥47.5 kg/m.
MATERIALS AND METHODSA total of 272 patients from Singapore university hospital who underwent LSG from 2008 to 2015 with a follow-up of at least 6 months were included in the study. Primary endpoint was weight loss at 1-year and 3-years. Morbid obesity (Group 1, G1) was defined as BMI <47.5 kg/m and super obesity (Group2, G2) was defined as BMI ≥47.5 kg/m.
RESULTSThere were 215 patients in G1 and 57 patients in G2 (mean preoperative weight: 107.3 kg and 146.8 kg; mean follow-up: 27.9 and 26.8 months, respectively). Mean total weight loss at 3-year of 41.9 kg for G2 was significantly higher ( = 0.003) than 27.2 kg for G1. Mean percentage excess weight loss (EWL) did not differ at 3-years. There was no difference in operating time, blood loss, length of stay, 30-day morbidity and readmission. There were no conversions and mortality in both groups. Remission of herpertension ( - 0.001) and dyslipidaemia ( = 0.038) were significantly associated with achieving EWL percentage (%EWL) >50 in G1.
CONCLUSIONLSG is an equally safe and effective operation in Asians with BMI ≥47.5 kg/m2 when compare to patients with BMI <47.5 kg/m in achieving significant weight loss and improvement in comorbidities. Super obese lose more weight but have lower %EWL.
Asian Continental Ancestry Group ; Gastrectomy ; methods ; Humans ; Laparoscopy ; instrumentation ; methods ; Obesity, Morbid ; surgery ; Outcome Assessment (Health Care) ; Patient Safety ; Retrospective Studies ; Singapore
8.Awake craniotomy of gliomas in eloquent areas: an analysis of 19 cases
Xi CHEN ; Liang ZHANG ; Jinli SUN ; Weichao JIANG ; Sifang CHEN ; Guowei TAN ; Zhanxiang WANG ; Hongming BAI
Chinese Journal of Neuromedicine 2017;16(12):1270-1274
Objective To discuss the techniques and methods of surgery for brain gliomas located in eloquent areas at awake anesthesia. Methods Nineteen patients with brain gliomas in eloquent areas, admitted to our hospital from December 2014 to May 2017, were operated under awake anesthesia with neuronavigation and intraoperative ultrasonography for locating the lesions and intraoperative direct electrical stimulation for functional mapping of the eloquent areas. All patients were followed up from 3 to 18 months; the surgical efficacies were analyzed. Results Of 19 patients, 18 (94.74%) were achieved awake and alert during brain mapping and resection of the tumors;17 (89.47%) were detected the motor areas by intraoperative direct electrical stimulation, 6 (31.58%) were detected the sensory cortex and 12 (63.16%) were detected language related cortex. Of 19 patients, MR imaging 2-3 months after surgery indicated that 5 (26.32%) received total resection of lesions, 9 (47.37%) subtotal resection of lesions and 5 (26.32%) partial resection of lesions. Seven patients (36.84%) had transitory postoperative aphasia, 4 (21.05%) were with transitory postoperative dyskinesia and one (5.26%) with permanent dyskinesia. Conclusion Comprehensive applications of awake anesthesia, neuronavigation, intraoperative ultrasonography and intraoperative direct electrical stimulation technologies allow maximum safe resection of gliomas in eloquent areas and protection of brain function.
9.Role of 3D printing positioning guide in neurosurgery
Zhongjie SHI ; Xin GAO ; Liwei ZHOU ; Shuo WANG ; Jianfeng GUO ; Guowei TAN ; Zhanxiang WANG
Chinese Journal of Neuromedicine 2021;20(10):1039-1042
Objective:To explore the application value of individualized three-dimensional (3D) printing positioning guides in localization and resection of intracranial lesions.Methods:Fifteen patients with intracranial space occupying lesions underwent resection in our hospital from March 2021 to May 2021 were selected in our study. Brain images by CT and MRI as raw data were used to design individual positioning guides. The positioning guides were placed on the patient's skin before resection to mark the location and boundary of the lesions with a marker, and neuro-navigation was used to verify the accuracy. During the resection, the location of the lesions was identified through microscope by the surgeons. Postoperative CT and MRI were used to evaluate the lesion resection.Results:The individualized positioning guides of 15 patients fit the skin well, and the skin incision and bone window were designed to meet the surgical requirements. All surgeries were completed in one time, and the lesion tissues were successfully removed. During the surgeries, the skin incision was not adjusted for secondary expansion. Brain MRI reexamination within 48 h of surgery showed that the lesions of 11 patients with tumors were removed satisfactorily (total resection in 9 and subtotal resection in 2); brain CT reexamination within 12 h showed that the clearance rate of hematomas in 3 patients was above 80% and that in 1 patient was 70%. No patients had cerebrospinal fluid leakage, intracranial hematoma, intracranial infection or other serious complications. All patients recovered well during the 1-3 months of outpatient/telephone follow-up.Conclusion:The positioning method with personalized 3D printing guides is simple and convenient, enjoying accurate positioning results, which can assist the clinicians to optimize the preoperative planning, optimize the surgical incision design, and is worthy of promotion and application in primary hospitals.
10.Effect of p38 mitogen-activated protein kinase pathway on hydrocephalus and aquaporin 4 expression in rats
Zhangyu LI ; Xin GAO ; Jianyao MAO ; Sifang CHEN ; Guowei TAN ; Zhanxiang WANG
Chinese Journal of Neuromedicine 2022;21(4):333-340
Objective:To observe the regulatory effect of p38 mitogen-activated protein kinase (p38 MAPK) on aquaporin 4 (AQP4) in rats after hydrocephalus, and to explore its significance in hydrocephalus prevention.Methods:Fifty SD rats were randomly divided into sham-operated group ( n=10), hydrocephalus group ( n=20), and hydrocephalus+inhibitor (SB203580) group (SB group, n=20). The rat models of hydrocephalus in the latter two groups were prepared by intracerebroventricular injection of kaolin suspension; rats in the sham-operated group were injected with same amount of normal saline into the lateral ventricle. The p38 MAPK specific inhibitor SB203580 (10 mg/kg) was intraperitoneally injected into the rats of SB group on the 8 th d of modeling for 7 consecutive d; same volume of dimethylsulfoxide was given to the rats of hydrocephalus group on the 8 th d of modeling for 7 consecutive d; rats in the sham-operated group did not give any treatment. The severity of hydrocephalus in these rats was observed by MRI. The inflammatory factor tumor necrosis factor (TNF)-α level in the cerebrospinal fluid was detected by enzyme-linked immunosorbent assay (ELISA). The AQP4 and TNF-α mRNA expressions were detected by real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR). The phosphorylated p38 MAPK and AQP4 expressions in the periventricular brain tissues were detected by Western blotting and immunohistochemistry. Results:No hydrocephalus developed in sham-operated group and hydrocephalus developed in the latter two groups. As compared with sham-operated group, hydrocephalus group and SB group had significantly increased lateral ventricle volume, significantly aggravated periventricular edema, significantly higher EVAN's index, and statistically increased brain water content ( P<0.05). Two weeks after modeling, the TNF-α expression levels in cerebrospinal fluid of sham-operated group, hydrocephalus group and SB group were (20.49±0.96), (42.04±3.17), and (28.00±3.71) pg/mL, respectively, with significant differences ( F=186.000, P<0.001); the TNF-α expression level in SB group was significantly higher than that in sham-operated group and significantly lower than that in hydrocephalus group ( P<0.05). Two weeks after modeling, the TNF-α and AQP4 mRNA expression levels in brain tissues of the three groups were significantly different ( P<0.05); the TNF-α and AQP4 mRNA expression levels in hydrocephalus group were significantly higher than those in sham-operated group and SB group ( P<0.05). Correlation analysis showed that there was a positive linear correlation between AQP4 mRNA expression and TNF-α mRNA expression in hydrocephalus group ( r=0.511, P=0.026), and there was a positive linear correlation between AQP4 protein expression and phosphorylated p38 MAPK protein expression in hydrocephalus group and SB group ( r=0.560, P=0.013; r=0.463, P=0.030). Immunohistochemical staining results showed that AQP4 expression was abundant in glial cells of the three groups; the p38 MAPK distribution was uniform and non-polar; the phosphorylated p38 MAPK protein expression in the hydrocephalus group was significantly higher than that in the sham-operated group, and that in the SB group returned to the level of the sham-operated group. Conclusion:The p38 MAPK pathway is involved in the positive regulation of AQP4 expression, which could be inhibited by SB203580.