1.Construction and screening of plasmid expression vectors containing short hairpin RNA targeting at vascular endothelial growth factor gene of GBC-SD cells
Huawei QU ; Yangde ZHANG ; Yuxiang CHEN ; Jiantai HE
Chinese Journal of Tissue Engineering Research 2009;13(50):9879-9882
BACKGROUND: Previous reseamh has proved that RNA interference can inhibit vascular endothelial growth factor (VEGF) gene expression of colon carcinoma, carcinoma of prostate, and retinoblastoma. However, RNA interference inhibiting VEGF of carcinoma of gallbladder was not reported. OBJECTIVE: To construct a plasmid expression vector coding for the short hairpin RNA (shRNA) targeting hVEGF165 mRNA. DESIGN, TIME AND SETTING: A gene engineering study was performed at National Hepatobiliary & Enteric Surgery Research Center, Xiangya Hospital, Central South University from 2008 to 2009.MATERIALS: Human GBC-SD was provided by Tumor Research Institute of Tongji University. METHODS: Four pairs of shRNAs that targeted at VEGF gene were designed. The eukaryotic expression plasmids (named shRNA1-4) were constructed and identified using restriction enzyme analysis. The plasmids were then transfected into GBC-SD cells via liposome2000. The transfection rate of recombinant plasmids was measured at 48 hours after transfection. MAIN OUTCOME MEASURES: Enzyme analysis of recombinant plasmid; transfection rate; VEGF mRNA expression determined using fluorescent polymerase chain reaction. RESULTS: shRNA plasmid vector targeting at VEGF gene was successfully constructed, in particular, pDC316-EGFP-U6-shRNA2 was the most effective. The expression plasmids were confirmed by restriction enzyme analysis. The transfection rate of recombinant plasmids in GBC-SD cells was approximately 58.6%. shRNA could inhibit VEGF mRNA expression, in particular, the inhibitory rate of RNA2 was the highest by 86%.CONCLUSION: The shRNA eukaryotic expression plasmid targeting at VEGF gene is constructed and selected successfully, and it can remarkably inhibit VEGF expression of GBC-SD cells. Additionally, the inhibitory effect of RNA2 is the greatest.
2.Small interfering RNA targeting vascular endothelial growth factor gone inhibits proliferation of gallbladder cancer cells: An in vitro and in vivo experiment
Huawei QU ; Yangde ZHANG ; Yuxiang CHEN ; Jinfeng ZHAO ; Mingmei LIAO ; Jiantai HE
Chinese Journal of Tissue Engineering Research 2010;14(2):258-261
BACKGROUND: Previous studies demonstrated that proliferation of cancer cells can be inhibited via RNA interference on the expression of vascular endothelial growth factor (VEGF). However, few studies report RNA interference on the expression of VEGF in gallbladder carcinoma, OBJECTIVE: To design and screen shRNA targeting VEGF, and to observe the effect of small interfering RNA targeting on proliferation of gallbladder cancer cells. METHODS: The VEGF-shRNA fragment was synthetized and connected with pCYU6/GFP/Neo-shRNA plasmid vector, shRNA was transfected into gallbladder cancer cells. The gallbladder carcinoma models of nude mice were prepared and randomly divided into blank control, negative control and experimental groups, With 6 animals in each group. ShRNA was injected into tumor. Cell growth was detected by fluorescence microscope MTT. The RNA interference efficiency was examined by fluorescent quantitative RT-PCR. Changes of tumor volume were also observed. RESULTS AND CONCLUSION: Gallbladder cancer cells ware shrunk with round shapes and a part of cells were dead after RNA interference on VEGF. shRNA-VEGF1 and shRNA-VEGF2 could signiticently inhibit mRNA gene expression of VEGF, the inhibition ratio was 86% and 82%, respectively. The tumor volume of the experimental group was smaller than the other groups, with slowly growth (P < 0.05). No obvious changes were found in the blank control and negative control groups. The constructed hVEGF-shRNA vector markedly decreases VEGF gene expression and inhibits cellular proliferation, eventually, to treat gallbladder cancer.
3.Analyses of the changes of sphenoid sinus and related factors of sphenoid sinusitis after microscopic transsphenoidal pituitary adenoma resection
Yong QIN ; Huawei WU ; Shanwu WU ; Hongyan QU ; Kunzhe LIN ; Yehuang CHEN ; Shousen WANG
Cancer Research and Clinic 2021;33(1):48-52
Objective:To investigate the changes of sphenoid sinus and related risk factors of sphenoid sinusitis after microscopic transsphenoidal pituitary adenoma resection.Methods:The clinical and imaging data of 106 patients with large pituitary adenoma in 900 Hospital of the PLA Joint Logistics Team between August 2012 and March 2015 were continually collected. The changes of accumulated blood and fluid, inflammation, mucocele, mucosa remodeling in sphenoid sinus cavity at preoperative and postoperative different time points were observed through the analysis of magnetic resonance imaging (MRI). Binary logistic multiple factors regression model was used to analyze the independent risk factors for postoperative sphenoid sinusitis.Results:MRI results showed that the blood and fluid accumulated in sphenoid sinus cavity were absorbed and dissipated 3 months after the surgery, and the saddle bone windows were covered by new mucous membrane at this time, but they were not complete; the remaining tumors in the saddle all sank into the saddle to different degrees. The reconstruction of sphenoid sinus mucosa was basically complete 6 months after the surgery. There were 7 (6.6%) cases of mucocele in sphenoid sinus and 26 (24.5%) cases of sphenoid sinusitis 3 months after the surgery among 106 patients. The results of multivariate analysis showed that growth hormone adenoma ( OR = 2.981, 95% CI 1.480-26.207, P = 0.014), preoperative sphenoid sinusitis ( OR = 12.392, 95% CI 2.927-52.462, P = 0.001), frequency of multiple transsphenoidal surgery ( OR = 14.758, 95% CI 2.431-89.584, P = 0.003) and perioperative cerebrospinal fluid leakage ( OR = 11.644, 95% CI 2.175-62.344, P = 0.004) were independent risk factors for postoperative sphenoid sinusitis. Conclusions:The evolution of sphenoid sinus cavity contents has its own rules after microscopic transsphenoidal pituitary adenoma resection. Patients with growth-hormone pituitary adenoma, sinusitis before surgery, multiple transsphenoidal surgery and cerebrospinal fluid leakage during the surgery should receive enhanced anti-infection treatment and nasal care in perioperative period to reduce the possibility of sphenoid sinusitis after surgery. The staged time of reoperation for pituitary adenoma resection by transsphenoidal approach should be about 3 months after the previous operation.
4.Effect of S-1 combined with oxaliplatin in the advanced pancreatic cancer and the level of immunocyte
Tao ZHANG ; Xin WANG ; Fengliang WANG ; Zhenxue CAO ; Huawei QU ; Wen PAN ; Changren LIU ; Yaning QUAN
Cancer Research and Clinic 2019;31(2):109-112
Objective To investigate the clinical effect of S-1 combined with oxaliplatin (SOX regimen) in treatment of the patients with advanced pancreatic cancer. Methods A total of 106 patients with advanced pancreatic cancer in Qingdao Fuwai Hospital from April 2015 to June 2017 were randomly divided into the treatment group (53 cases) and the control group (53 cases) according to the random number table method. Patients in the control group were treated with S-1 combined with cisplatin treatment, and patients in the treatment group were treated with SOX regimen. The cell proportion of CD3+, CD4+, CD8+and CD4+/CD8+before treatment and after 2 cycles of treatment were detected by using flow cytometry of both groups. The clinical curative effects, immunity and adverse reactions of both groups were compared by usingχ2 test and t test. Kaplan-Meier method was used to make the survival analysis. Results After two cycles of treatment, there were 4 cases of complete remission (CR), 23 cases of partial remission (PR), 17 cases of stable disease (SD), 9 cases of progression disease (PD) in the treatment group, and 0 case of CR, 18 cases of PR, 20 cases of SD, 15 cases of PD in the control group. The rate of CR+PR in the treatment group was higher than that in the control group [50.94%(27/53) vs. 33.96%(18/53)], and there was a statistical difference (χ2=5.936, P<0.05). There was a significant difference in the cell proportion of CD4+and ratio of CD4+/CD8+between the two groups before and after treatment [the treatment group: (27.31±2.48)% vs. (37.05±2.53)%, χ2= 6.491,P< 0.01; 0.91 ±0.23 vs. 1.53 ±0.50, χ2 = 5.913, P< 0.01; the control group: (27.43 ±2.47)% vs. (30.32 ± 2.41)%,χ2= 11.214, P<0.01; 0.90±0.22 vs. 1.22±0.34,χ2=7.992, P<0.01]. After 2 cycles of treatment, the cell proportion of CD4+and ratio of CD4+/CD8+of the treatment group were higher than those of the control group, and there were statistical differences (χ2=5.309, P<0.01;χ2= 7.112, P< 0.01). The incidence rate of side effects had no significant difference in both groups after two cycles of treatment [22.64% (12/53) vs. 18.87% (10/53), χ2= 1.924, P> 0.05]. The progression-free survival time in the treatment group was longer than that in the control group (P<0.05). Conclusions SOX regimen has a favorable effect on the patients with advanced pancreatic cancer. It can help to improve the immunity and prolong the survival time of the patients.
5.Progress and confusion in the diagnosis and treatment of scar
Jiarong YI ; Honglian ZHANG ; Jia CHEN ; Siwei QU ; Huawei OUYANG ; Chaoqi YIN ; Bin HE ; Ke TAO ; Jianda ZHOU
Journal of Chinese Physician 2019;21(4):481-484
The prevention and treatment of scars has always been an important task in the field of wound repair.Humans have started treatment for scars long time ago and have created a series of treatments.At the same time,with the development of science and technology,the diagnosis and treatment of scars have made great progress.Through the basic research for new therapeutic targets and innovative of treatment methods,treatment for scar turns more diversified and integrated.As a result,clinicians will suffer many challenges while having more options for scars treatment.Combined with traditional treatment,developing new treatments,and the comprehensive therapy of multiple treatment methods is the mainstream trend of scar treatment.