1.A study on the expressions of IL-17A and IL-17RA in different degrees of malignant glioma
Lei WANG ; Yanbo LIU ; Zhenjiang WANG ; Xinli ZHAO ; Weigao SHEN
Tianjin Medical Journal 2016;44(8):1018-1021
Objective To explore the expressions of interleukin (IL)-17A and its receptor IL-17RA in different degrees of malignant gliomas. Methods Fifty patients with glioma were collected in this study. Accordance to the World Health Organization Classification System, patients were classified by malignancy grade, including gradeⅠ(n=12), gradeⅡ(n=18), gradeⅢ(n=13) and gradeⅣ(n=7). The glioma tissue and peripheral blood samples of patients were obtained for detecting the expression levels of IL-17A and IL-17RA mRNA by using immunohistochemistry, quantitative real-time PCR. Western blot assay was used to detect expressions of IL- 17A and IL- 17RA in both the macroscopic (immunohistochemistry) and molecular levels (mRNA and protein). Results Immunohistochemical staining showed that the expression levels of IL-17A and its receptor IL-17RA increased with the increase of the malignant degree of gliomas. The mRNA levels of IL-17A and IL-17RA receptors in peripheral blood were up-regulated with the increasing malignancy grade of glioma (F=8.96, P<0.05;F=10.34, P<0.05). The mRNA levels of IL-17A and IL-17RA in glioma tissues were up-regulated with the increasing malignancy grade of glioma (F=11.21, P<0.05;F=14.11, P<0.05). The protein levels of IL-17A and IL-17RA in peripheral blood and glioma tissues were also up-regulated with the increasing malignancy grade of glioma (in peripheral blood:F=9.90, P<0.05;F=11.80, P<0.05;and in gliomas tissues:F=8.15, P<0.05;F=14.46, P<0.05). Conclusion The expressions of IL-17A and IL-17RA receptor are positively correlated with malignancy grade of glioma. These results provide some reference for clinical diagnosis of malignant gliomas.
2.Effects of fasudil on the expressions of Bcl-2 and Bax in cerebral cortex of rats with subarachnoid hemorrhage
Chunming MIAO ; Weigao SHEN ; Lei WANG ; Dun WEI
Tianjin Medical Journal 2016;44(12):1436-1439,1440
Objective To investigate the effects of fasudil on expressions of Bcl-2 and Bax in cerebral cortex of model rats with subarachnoid hemorrhage (SAH). Methods Thirty rats were divided into sham operation group, SAH group and SAH+fasudil group, 10 rats in each group. Double injection of blood into occipital cistern method was used for SAH model in SAH group and SAH+fasudil group. In the sham operation group, the blood injection was instead by normal saline. In the SAH+fasudil group, at 30 min after the second injection of blood, rats were administrated with intraperitoneal injection of fasudil at a dose of 3 mg/kg. The general situation, neurological score, TUNEL staining for cortex cell apoptosis, immune histochemical staining and Western blotting assay for Bcl-2 and Bax protein expression were compared 24 h after the operation between the three groups. Results Compared with the sham operation group, rats in SAH group and SAH +fasudil group appeared obvious neurological deficits. The neurological score was significantly lower in SAH group ( 2.68 ± 0.31) than that of sham operation group (5.00±0.00). The neurological score was significantly higher in SAH+fasudil group (3.27 ± 0.35) compared with that of SAH group (2.68 ± 0.31, P<0.05). There was obvious cell apoptosis in SAH group and SAH+fasudil group, and the apoptosis was less in SAH+fasudil group than that of SAH group (P<0.05). The level of Bcl-2 expression was significantly lower in SAH group than that of sham operation group, and Bax expression was significantly higher in SAH group than that of sham operation group (P<0.05). The level of Bcl-2 expression was significantly higher in SAH+fasudil group than that of SAH group, but Bax expression was significantly lower in SAH+fasudil group than that of SAH group (P<0.05). Conclusion Fasudil can improve the neurological damage in rats with SAH, which may be related with the regulation of apoptosis related proteins Bcl-2 and Bax.
3.Changes of BDNF level expression in Hippocampal and Habenular nucles in experimental depression rats
Wei XIA ; Hua ZHAO ; Aihua REN ; Weigao SHEN
Chinese Journal of Immunology 1986;0(04):-
Objective:To observe the change of BDNF level expression in experimental in hippocampal and habenular nucles in experimental depression rats.Methods:The depression model of rats was established by chronic unpredictable mild stress. Openfield test was performed to detect the behavior of rats. Immunohistochemistry was used to observe the changes of BDNF level.Results:Compared to the control group,the number of depressed animals was much less than that of normal animals.Conclusion:The level of BDNF decrease significantly in experimental depressive model of rats.
4.Application of laparoscopic partial nephrectomy with individual operation plan in treatment of small renal cell carcinoma
Sihai SHAO ; Jianer TANG ; Yuefan SHEN ; Anping XIANG ; Weigao WANG ; Rongjiang WANG
China Journal of Endoscopy 2017;23(8):105-110
Objective To evaluate the feasibility and clinical efficacy of retroperitoneal laparoscopic partial nephrectomy (RLPN) with individual operation plan in treatment of small renal cell carcinoma (RCC). Methods 98 patients with small RCC who was treated by RLPN from June 2012 to June 2016 were retrospectively analyzed. There were 57 males and 41 females with a mean age of 52 years old (ranging 28 ~ 75 years old). 52 cases were located on the right side while 46 cases were left. The mean tumor size was 3.1 cm in diameter (ranging 0.8 ~ 4.5 cm). 87 patients (A group) were underwent standard RLPN with clamping main renal artery. 7 patients (B group) with exophytic RCC were performed without clamping renal artery, but with separating main renal artery and prepared for possible clamping. 4 patients (C group) with endophytic RCC were performed with clamping renal artery under ultrosound monitoring. The feasibility and outcomes were evaluated by surgical and oncological outcomes. Results 84 cases among A group were underwent standard RLPN successfully, with 2 cases converted to open surgery and 1 case failed to excising tumor completely and converted to laparoscopic radical nephrectomy. The amount of bleeding during operation was 30 ~ 350 ml, average 93 ml, operation time was 70 ~ 245 min, average 127 min, warm ischemia time 20 ~ 42 min, average 26 min. 6 cases among B group were performed successfully without clamping renal artery with 1 case converted to clamp renal artery for 15 min during the operation because of obvious bleeding. The amount of bleeding was 160 ~ 380 ml, average 220 ml, operation time was 85 ~ 215 min, average143 min. 4 cases of C group were all performed successfully, The amount of bleeding was 35 ~ 250 ml, average 85 ml, operation time was 110 ~ 235 min, average 175 min, warm ischemia time 25 ~ 40 min, average 28 min. With a mean follow up of 28 months (ranging 18 ~ 42 months), there was only 1 case of A group occured local recurrence and lung metastases and accepted molecular targeted therapy with Sorafenib. Conclusion RLPN with individual operation plan in treatment of small RCC is safe and effective, the long-term effect of the procedure needs further investigation.
5.Risk factors analysis of persistent frequency after transurethral resection of the prostate
Junwen SHEN ; Fuding BAI ; Chuanjun DU ; Weigao WANG
Chinese Journal of Urology 2018;39(1):34-37
Objective To discuss the risk of factors influencing persistent frequency after transurethral resection of the prostate (TURP).Methods The clinical data of 119 post-TURP patients treated from January 2014 to June 2015 was retrospectively analyzed.The age was (72.1 ±2.3)years old.There were 15 cases with hypertension,23 cases with diabetes and 6 cases with heart disease.The preoperative IPSS score of 119 cases was (22.1 ± 5.9) points,with (10.2 ± 1.8) points in urinary storage period and (11.8 ± 4.7) points in urination period.Urination frequency was (10.8 ± 2.6) times per day and there were (3.8 ± 0.8) times of nocturnal urination.B-ultrasound:residual urine volume was (38.1 ± 9.1) ml and prostate volume was (34.1 ± 4.2) ml.Preoperative maximum urine flow rate was (8.8 ± 3.9) ml/s.The detrusor pressure at maximum urinary flow rate was (43.9 ± 14.1) cm H2O (1 cmH2O =0.098 kPa),maximum detrusor pressure was (99.7 ± 12.2) cmH2O and effective bladder volume was (217.5 ± 14.8) ml.Contraction of bladder weakened in 40 cases (33.6%) and 36 cases (30.2%) had detrusor overactivity.According to whether continuous urinary frequency was developed,the patients were divided into frequency-positive group and frequency-negative group.The differences between the patients in two groups were compared and univariate analysis was performed.A multivariate logistic regression was performed on statistically significant indicators.Results Among the 119 patients,21 were frequency-positive and 98 were frequency-negative.Univariate analysis showed that age,IPSS score,preoperative urinary storage score,detrusor pressure at maximum urinary flow rate,maximum detrusor pressure,effective bladder volume,contraction decrease of bladder,preoperative detrusor activity were important indicators affecting the condition of postoperative urinary frequency (all P < 0.05).Multivariate analysis showed that old age (OR =3.842,P =0.021),high total IPSS score (OR =5.109,P =0.011),low maximum detrusor pressure (OR =3.477,P =0.039),low effective volume of bladder (OR =4.051,P =0.017) and detrusor overactivity (OR =3.662,P =0.025) were independent risk factors for urinary frequency after TURP.Conclusions The age,the high IPSS score before operation,low maximal detrusor pressure,low effective bladder capacity and the bladder detrusor activity could be independent predictive factors of persistent frequency after TURP.
6.Over-expression of programmed cell death 5 gene enhances chemosensitivity of brain glioma cells to temozolomide
WANG Lei ; SHEN Weigao ; LIU Yanbo ; XU Haiyang
Chinese Journal of Cancer Biotherapy 2019;26(8):868-875
Objective: To explore the role of tumor suppressor gene programmed cell death 5 gene (PDCD5) in the growth and temozolomide (TMZ) sensitivity of brain glioma cells. Methods:Atotal of 116 patients with cerebral glioma admitted to the Department of Neurosurgery, First Clinical Hospital of Jilin University from January 2009 to December 2014 were enrolled in this study. QPCR, WB and immunohistochemistry method were used to detect the mRNAand protein expressions of PDCD5 in glioma cell lines (U87, U251), U87 cell line with stable PDCD5 expression (U87-PDCD5), glioma cells with si-PDCD5 transfection and primary cerebral glioma tissues, respectively. MTT assay was used to detect the effect of over-expression or knockdown of PDCD5 on the growth and TMZ-sensitivity of glioma cells. The subcutaneous tumor-bearing model of glioma cell line U87 was established in nude mice, and then the experimental mice were randomly divided into control group, TMZ group, PDCD5 group and TMZ+exogenous PDCD5 recombinant expression vector group.After 20 days, the animals were sacrificed by cervical dislocation and the tumor tissue was excised to measure the tumor volume and weigh. The expression of PDCD5 in tumor tissues was detected by qPCR and WB methods, and the effects of PDCD5 combined with TMZ on the growth of gliomas were also analyzed. Results: The relative mRNA and protein expressions of PDCD5 in U87 cells were significantly lower than those in U251 cells (both P<0.05), and the mRNA and protein expressions of PDCD5 in high level glioma tissues were significantly lower than those in low level tissues (all P<0.05). The sensitivity of U87-PDCD5 cells and U251 cells to TMZ was higher than that of U87 cells (all P<0.05). The sensitivity of cells to TMZ in U87-PDCD5-siRNA group and U251siRNA group was significantly lower than that of the control group (both P<0.05). The tumor volume and weigh to fnudemicexenografts were compared,and the results showed control group>TMZ group>PDCD 5group>combined group(allP<0.05);however, the mRNA and protein expressions of PDCD5 in the transplanted tumor tissues of each group showed the opposite trend (all P<0.05). Conclusion: PDCD5 over-expression can enhance the chemosensitivity of braingliomato the chemotherapy drug TMZ, while silencing of PDCD5 expression exertsthe opposite effect.The combination of PDCD5 and TMZ can better inhibit the growth of xenografts in nude mice.
7.miR-423-5p enhances chemoresistance of glioblastomas cells to temozolomide by regulating PDCD5
WANG Lei ; SHEN Weigao ; LIU Yanbo
Chinese Journal of Cancer Biotherapy 2020;27(10):1100-1105
[Abstract] Objective: To investigate the miR-423-5p expression in brain glioma tissues and cell lines, and its promotive effect on
temozolomide (TMZ) chemoresistance by targeting PDCD5 (programmed cell death protein 5). Methods: Tumor tissues and matched
peritumoral tissues were collected from 20 brain glioma patients who were surgically treated in the Department of Neurosurgery,
Affiliated Hospital of Beihua University between January 2017 and December 2018. Glioblastoma cell lines (U251, U87, SHG-44)
and human normal glial cell line HMC-3 were also used in the study. The relative expression of miR-423-5p and PDCD5 in brain
glioma and peritumoral tissues and cell lines was detected by qPCR. The synthesized miR-423-5p mimics and miR-NC were respectively
transfected into U251 and U87 cells; meanwhile, TMZ at different concentrations (50, 100, 150 and 200 μmol/L) were also used to treat
the cells. Then, the chemoresistance of cells to TMZ were determined. MTT assay and colony formation assay were used to examine the
proliferation of U251 and U87 cells, andWestern blotting was used to detect the expression of c-caspase 3, Bcl-2 and PDCD5 proteins in
U251 and U87 cells. The targeting relationship between PDCD5 and miR-423-5p was validated through Dual luciferase reporter gene
assay. Results: miR-423-5p was highly expressed in glioma tissues and glioma cell lines (all P<0.01). As compared with the miR-NC
group, the proliferation and TMZ-chemoresistance of U251 and U87 cells in miR-423-5p mimics group significantly increased (all
P<0.01). Dual luciferase reporter gene assay validated that miR-423-5p could bind with PDCD5 3' UTR to suppress the expression of
PDCD5. Conclusion: High expression of miR-423-5p enhances the chemoresistance of glioma cells to TMZ, and miR-423-5p may
serve as a potential therapeutic target in the treatment of brain glioma.