1.Effect and mechanism of TIPE3 interference plasmid on SW480 colorectal canc-er growth
Yuhan YE ; Zhongchen LIU ; Zhongquan QI ; Guohong ZHUANG ; Ping YIN
Chinese Journal of Immunology 2017;33(3):378-383
Objective:To study the effect of interference TIPE3 on the colon cancer cell growth by transfecting SW480 colon cancer cells with the TIPE3 interference plasmid were detected.Methods:Transfecting the constructed TIPE3-shRNA-pSIREN-RetroQ plasmid to SW480 cells.To determine the highest interference efficiency plasmid ,the mRNA and protein levels of recombined plasmid were detected by RT-PCR and Western blot separately and tested the cell proliferation with CCK 8.Meanwhile,apoptosis rate of SW480 cells was determined by flow cytometry assay with AnnexinV-FITC/PI.To further determined the effects of recombined plasmid on cell development ,the level of protein involved in proliferation and apoptosis were detected by Western blot .Results:The most effecient in-terference plasmids were successfully constructed.We found that the cell survival rate decreased when interference TIPE 3 gene express-ing in colorectal cancer cells .Flow cytometry indicated that interefering the expression of TIPE 3 would increase the sensitivity of SW 480 cell to apoptosis induced by aDR5ScFv.The results of Western blot showed that low expression of TIPE 3 would activate caspase3 and downregulate the expression of p-AKT,p-PDK1 and PCNA.Conclusion:Interference TIPE3 could promote apoptosis and inhibit prolif-eration in SW480 colon cancer cells .
2.AMACR (P504s)—a new immunohistochemical method for diagnosis of prostate carcinoma
Lu SHENG ; Jianda SONG ; Qi GU ; Li XIAO ; Zhongquan SUN ; Weili QIAN
Chinese Journal of Urology 2001;0(06):-
Objective To evaluate the expression of alpha-methylacyl-coenzyme-A racemase (AMACR,P504s) with immunohistochemical staining in diagnosis of prostate carcinoma. Methods A total of 133 specimens were taken from the patients (mean age,71 years),including 46 cases of prostate carcinoma (PCa) (1 case of stage A,19 cases of stage B,14 cases of stage C,12 cases of stage D;4 cases of gradeⅠ,14 cases of gradeⅡ,28 cases of grade Ⅲ),53 cases of benign prostate hyperplasia (BPH),13 cases of prostate intraepithelial tumor (PIN),9 cases of normal prostate,6 cases of prostatitis,3 cases of metastatic prostate cancer.Two sections of each specimen were made,with 1 stained by hematoxylin and eosin,and the other stained immunohistochemically by a rabbit monoclonal antibody to AMACR (P504s).AMACR staining expression was characterized as negative (score,1), weak (positive) (2), moderate (3) or strong (4). Results In 46 cases of PCa,AMACR staining expression was negative in 2 cases,weak in 1,moderate in 25 and strong in 18,with a mean staining intensity of 3.28 [95% confidence interval (CI), 3.07-3.50]. In 53 cases of BPH, the staining expression was negative in 47 cases, weak in 6, with a mean intensity of 1.11 (95% CI, 1.02-1.20).In 13 cases of PIN, the staining expression was negative in 12, weak in 1, with a mean intensity of 1.08 (95% CI, 0.91-1.24).The score of PCa group was significantly different from those of the latter 2 groups (P
3.Therapeutic effect of different dosages of praziquantel on mice infected with Sparganum mansoni
Nan LI ; Ximeng LIN ; Jiang CUI ; Mingming WANG ; Fengjun JING ; Xin QI ; Li WANG ; Zhongquan WANG
Chinese Journal of Schistosomiasis Control 2010;22(1):51-55
Objective To observethe efficacy of mice infected with Sparganum mansoni by using different dosages of praziquantel.Methods A total of 156 Kunming mice were divided into 2 batches.each of them wag orally infted with 5 spargana.Thirty-six mice in the first batch were equally divided into 6 groups.the mice in group 1-5 were inoculated with spargana cultured in different concentrations of praziquantel for 3 days,the group 6 served as a control.One hundred and twenty mice in the second batch were equally divided into 12 groups,each mouse was inoculated with spargana obtained from frogs or tadpoles,group 1-9 were treated by different desages of praziquantel 1 or5 weeks post infection.group 10-12 served as controls.All of the mice wore sacriftced and dissectedl or 2 weeks after the treatment.the mean number of worms recovered was cmculated and worm reduction rates were determined.Results The number of worm recovered from mice infected with spargana cultured in 10-40 μg/ml of praziquantel had no significant difference with that of the control(P>0.05).The worm reduction rate wag 16.60%while the spargana beins cultured in 50 μg/ml of praziquantel.The worm reduction rates of the mice that sacrificed 1 week or 2 weeks after being treated by the same dosage of praziquantel had no significant difference(P>0.05).When being treated with 200.400 or 800 mg/kg of praziquantel 1 week post infection,the number of worm recovered from mice infected with spargana from frogs had no significant difference with those of the control 1 and 2 weeks after the treatment(P>0.05).The worm reduction rates between the groups with the same dosage 1 week and 2 weeks post treatment had no significant difference(P>0.05).When being treated with 200 or 400 mg/kg of praziquantel 1 week post infection,the number of worm recovered from mice infected with spargana from tadpoles had no statistically difference with that of the control 1 week after treatment (P>0. 05). The worm reduction rate of mice was only 17.02% while being treated with 800 mg/kg of praziquantel. The worm reduction rates among groups with different dosages had no significant difference (P>0.05). Compared with the mice infected with spargana from frogs treated with 1 200 or 1 800 mg/kg of praziquantel 5 weeks post infection, the difference between the numbers of worm recovered from mice 1 week and 2 weeks after treatment had no statistically significance (P > 0.05), but they were significantly higher than those of the controls (P<0.05 ). The worm reduction rates among the groups with the same dosage had no significant difference (P>0.05). ConclusionsPraziquantel (10-50 μg/ml) has no evident killing effect on spargnna in vitro, but when the dosage is higher(1 800 mg/kg), it has certain efficacy for treating the mice infected with spargana by oral inoculation.
4.Clinical Effect of Cetuximab Combined with Chemotheraphy on Metastastic Colorectal Cancer
Liping BAI ; Zhongchen LIU ; Zhongquan QI ; Zhijie DING ; Sibo YUAN ; Shifeng ZHANG ; Xingfeng QIU
Chinese Journal of Clinical Oncology 2010;37(4):220-222
Objective: To evaluate the efficacy and the adverse reactoions of cetuximab combined with cheomotherapy (oxapliplatin or iriticon) for metastastic colorectal cancer. Methods: A total of 22 patients with metastastic colorectal cancer were treated with cetuximab combined with FOLFIRI or mFOLFOX6. The patients received cetuximab at an initial dose of 400 mg/m~2 intravenously on day 1 in the first cycle, followed by weekly infusion of 250 mg/m~2; FOLFIRI: irinotecan 180 mg/ m~2 on day 1, CF 400 mg/m~2, 5-FU bolus 400 mg/m~2, 5-FU infusion 2400 mg/m~2 over 46 hours, once every 2 weeks; mFOLF-OX6: oxaliplatin 85mg/m~2 on day 1, CF 400 mg/m~2, 5-FU bolus 400 mg/m~2, 5-FU infusion 2400 mg/m~2 over 46 hours, once every 2 weeks. The immediate response, complete response and partial response and changes in tumor marker levels were observed. Results: There were 12 PR cases, 6 SD cases, and no CR cases. The rate of (CR+PR) was 57.1% and the rate of (CR+PR+SD) was 85.7%. The adverse reactions during the theraphy were skin toxicity and neutropenia. Conclu-sion: Safe and effective for metastastic colorectal cancer, cituximab combined with oxaliplatin or irinotecan can increase the resectabiliy rate and prolong patient survival.
5.Construction and identification of interference plasmid targeting on TNFAIP8
Wenming LIU ; Jingjing YANG ; Ruyi HU ; Xingfeng QIU ; Chunyan SHI ; Zhongquan QI ; Zhongchen LIU ; Guohong ZHUANG
Chinese Journal of Immunology 2015;(5):650-654
Objective:To construct and screen the high efficiency interference plasmid of TFAIP8-shRNA-pSIREN-RetroQ.Methods:Selected and synthesized three Target Sequence of TNFAIP8 shRNA1,TNFAIP8 shRNA2,TNFAIP8 shRNA3,and construct the TNFAIP8 interference plasmid.Transfection TNFAIP8-shRNA-pSIREN-RetroQ interference plasmid to A549 cells.Filter out the highest interference efficiency plasmid by detecting the mRNA and protein levels using RT-PCR and Western blot methods.Results:We successfully design and built three TNFAIP8-shRNA-pSIREN-RetroQ interference plasmids,and screen out the highest efficiency interference plasmid.Conclusion: Three interference plasmids targeting the TNFAIP8 gene have been constructed successfully and provide a useful tool for studying the function of TNFAIP8.
6.Complete mesocolic excision in laparoscopy-assisted right hemicolon carcinoma radical resection
Xingfeng QIU ; Zhijian YE ; Feng YAN ; Li LIN ; Sibo YUAN ; Zhijie DING ; Zhenfa WANG ; Zhongquan QI ; Zhongchen LIU
Chinese Journal of General Surgery 2012;27(3):213-215
Objective To evaluate complete mesocolic excision in laparoscopy-assisted right hemicolon carcinoma radical resection. Methods Laparoscopy-assisted right hemicolon carcinoma radical resection with complete right-side mesocolic excision was performed in 36 cases between June 2010 and July 2011 at Zhongshan Hospital,Xiamen University. Results The operations were completed successfully without conversion to open surgery.The mean operative time was (134 ±22) min.The blood loss was (95 ±53 ) ml.The median number of total lymph nodes removed was 15.7.The average time for passage of flatus was (3.1 ± 1.2) d.The postoperative complications were observed in 6 of 36 cases (17%) including lymphatic fistulas in 4 patients,pulmonary infection in 1 patient and postoperative bleeding in 1 case.Conclusions Laparoscopy-assisted complete right-side mesocohc excision can be successfully performed for right hemicolon carcinoma,and the lymphoid tissue could be eliminated maximally.The long-term results need further evaluation.
7.Homoharringtonine promotes heart allograft acceptance by enhancing regulatory T cells induction in a mouse model
Xia QIU ; Hedong ZHANG ; Zhouqi TANG ; Yuxi FAN ; Wenjia YUAN ; Chen FENG ; Chao CHEN ; Pengcheng CUI ; Yan CUI ; Zhongquan QI ; Tengfang LI ; Yuexing ZHU ; Liming XIE ; Fenghua PENG ; Tuo DENG ; Xin JIANG ; Longkai PENG ; Helong DAI
Chinese Medical Journal 2024;137(12):1453-1464
Background::Homoharringtonine (HHT) is an effective anti-inflammatory, anti-viral, and anti-tumor protein synthesis inhibitor that has been applied clinically. Here, we explored the therapeutic effects of HHT in a mouse heart transplant model.Methods::Healthy C57BL/6 mice were used to observe the toxicity of HHT in the liver, kidney, and hematology. A mouse heart transplantation model was constructed, and the potential mechanism of HHT prolonging allograft survival was evaluated using Kaplan–Meier analysis, immunostaining, and bulk RNA sequencing analysis. The HHT-T cell crosstalk was modeled ex vivo to further verify the molecular mechanism of HHT-induced regulatory T cells (Tregs) differentiation. Results::HHT inhibited the activation and proliferation of T cells and promoted their apoptosis ex vivo. Treatment of 0.5 mg/kg HHT for 10 days significantly prolonged the mean graft survival time of the allografts from 7 days to 48 days ( P <0.001) without non-immune toxicity. The allografts had long-term survival after continuous HHT treatment for 28 days. HHT significantly reduced lymphocyte infiltration in the graft, and interferon-γ-secreting CD4 + and CD8 + T cells in the spleen ( P <0.01). HHT significantly increased the number of peripheral Tregs (about 20%, P <0.001) and serum interleukin (IL)-10 levels. HHT downregulated the expression of T cell receptor (TCR) signaling pathway-related genes ( CD4, H2-Eb1, TRAT1, and CD74) and upregulated the expression of IL-10 and transforming growth factor (TGF) -β pathway-related genes and Treg signature genes ( CTLA4, Foxp3, CD74, and ICOS). HHT increased CD4 + Foxp3 + cells and Foxp3 expression ex vivo, and it enhanced the inhibitory function of inducible Tregs. Conclusions::HHT promotes Treg cell differentiation and enhances Treg suppressive function by attenuating the TCR signaling pathway and upregulating the expression of Treg signature genes and IL-10 levels, thereby promoting mouse heart allograft acceptance. These findings may have therapeutic implications for organ transplant recipients, particularly those with viral infections and malignancies, which require a more suitable anti-rejection medication.