1.Potential mechanism of Atractylodes macrocephala aqueous extract inhibiting gastric carcinoma through PI3K-Akt-NF-κB signaling pathway
HA Wentao ; ZHAO Sunyan ; WEI Xiaowei ; GONG Yongling
Chinese Journal of Cancer Biotherapy 2023;30(3):223-229
		                        		
		                        			
		                        			[摘  要]  目的:探讨白术(Atractylodes macrocephala)水提物抑制胃癌SGC-7901细胞活性的潜在机制。方法:分别使用蒸馏水(对照)和白术水提物(白术治疗组)灌胃SD大鼠后,采集静脉血后分离其血清、过滤并分别命名为对照组血清(CON-S)和白术组血清(AM-S)。将胃癌SGC7901细胞分为对照组、10% AM-S组和20% AM-S组,其中两个AM-S组细胞分别在相应浓度的AM-S血清中培养24 h,对照组细胞用正常培养基培养相同时间,收取SGC7901细胞和上清液用于进一步分析。使用MTT法检测各组细胞活力,通过商业试剂盒测定乳酸脱氢酶(LDH)、丙二醛(MDA)和超氧化物歧化酶(SOD)的水平,采用ELISA试剂盒检测各组细胞中IL-6和TNF-α的含量,采用WB法评估各组细胞中PI3K-Akt-NF-κB信号通路相关蛋白的表达。结果:10% AM-S组和20% AM-S组的SGC7901胃癌细胞增殖活力相较于对照组分别降低48.9%和53.25%(P<0.05或P<0.01);胃癌细胞上清液中,相较于对照组,10% AM-S组和20% AM-S组LDH水平分别升高29.25%和123%、SOD活性分别升高18%和54.60%、MDA水平分别降低27.8%和40.0%,IL-6水平分别降低15%和17.5%、TNF-α水平分别降低29.71%和40.16%(P<0.05或P<0.01)。相较于对照组,AM-S组中PI3K-Akt-NF-κB信号相关蛋白的水平显著下降(P<0.05或P<0.01)。结论:白术水提物可以通过抑制癌细胞增殖活力、促进凋亡、抑制肿瘤微环境中的促炎因子分泌以及改变细胞内的氧化应激水平等方式抑制胃癌,其机制可能是通过抑制PI3K-Akt-NF-κB通路来实现这些抗癌作用的。  
		                        		
		                        		
		                        		
		                        	
2.Successful treatment of 3 cases of 22-week extremely premature infants
Yanliang YU ; Wentao GONG ; Nian LIU ; Yongping FU ; Jie ZHAO ; Bingchun LIN ; Zhifeng HUANG ; Chuanzhong YANG
Chinese Journal of Neonatology 2022;37(5):448-451
		                        		
		                        			
		                        			Objective:To review the treatment experience of extremely premature infants (EPIs) with gestational age (GA) <23 weeks.Methods:From January to November 2021, EPIs with GA<23 weeks treated in our hospital was retrospectively analyzed.Results:A total of 3 patients with GA of 22 weeks were reviewed, including 2 boys and 1 girl. Their birth weight (BW) was 450~498 g. The duration of hospitalization was 112~126 d. The treatment included early "gentle" management strategies, respiratory management, anti-infection, patent ductus arteriosus treatment and parenteral + enteral nutrition. All 3 infants were discharged from the hospital without further oxygen therapy. All had satisfying oral feeding with no neurological sequelae on follow-up.Conclusions:Early "gentle" management is the key to successful treatment and good prognosis for EPIs with GA<23w
		                        		
		                        		
		                        		
		                        	
3.Function and mechanism of ferroptosis in the radiation resistance of colorectal tumor-repopulating cells
CHANG Yuhan ; Ge Yutong ; HA Wentao ; WEI Xiaowei ; GONG Yongling
Chinese Journal of Cancer Biotherapy 2022;29(5):426-433
		                        		
		                        			
		                        			[Abstract] Objective: To investigate the function and mechanism of ferroptosis in the radiation resistance of colorectal tumor-repopulating cells. Methods: Human colorectal tumor cells HCT116 (defined as Control cells) were cultured in two-dimensional normal conditions, and tumor regenerative cells with high tumorigenicity (defined as TRCs) were cultured and screened in three-dimensional fibrin soft gels by the mechanical force method. Both the control group and TRC group cells were exposed to X-rays with different doses (2, 4, 6, 8 Gy) and  MTS and the clone formation assay were used tomeasure the cell viability rate and proliferation ability. After the Control cells and TRCs were treated with ferroptosis inducer (Erastin) and X-rays respectively, they were stained with C11-BODIPY reagent, and the lipid peroxidation level of the cells was observed and determined by confocal microscopy and flow cytometry. qPCR was used to determine the effects of Erastin and X-rays treatments on the expressions of ferroptosis-related genes glutathione peroxidase 4 (GPX4) and acyl-coenzyme A synthetase long-chain family member 4 (ACSL4) in the Control cells and TRCs; WB assay was performed to determine the effects on the expressions of ferroptosis-related proteins GPX4 and ACSL4. Results: Colorectal TRCs with high stemness were cultured and screened out from soft fibrin gels. After irradiation with different doses (2, 4, 6, 8 Gy) of X-rays, the viability rate, the clone sizeand the number of clones in the control group were significantly lower than those in the TRC group (all P<0.05). After the cells in the control group were irradiated with different doses of X-rays (4, 8 Gy) and treated with Erastin, the lipid peroxidation level of the cells in the X-ray treated group was significantly higher than that in the untreated group (P<0.05). The lipid peroxidation level of the cells in the Erastin-treated group was significantly higher than that in the DMSO-treated group (P<0.05). There was no statistical difference among all treatment subgroups in the TRC group (all P>0.05). The mechanism study indicated that compared with those in control cells, GPX4 and ACSL4 in TRCs under ferroptosis-inducing conditions (X-ray radiation and Erastin treatment) presented expressions that contributed more to radiation resistance, i.e., continued upregulation of GPX4 and downregulation of ACSL4 and their expressions were dependent on the doses of Erastin. Conclusion: Colorectal TRCs may resist ferroptosis through a high expression of GPX4 and a low expression of ACSL4, which in turn induces radiation resistance.
		                        		
		                        		
		                        		
		                        	
4.Impulsive-like behaviors of rats in Y-maze task induced by pramipexole and its mechanism
Wentao MA ; Xiang GAO ; Zehui GONG ; Zheng YONG ; Ruibin SU
Chinese Journal of Pharmacology and Toxicology 2016;30(5):491-497
		                        		
		                        			
		                        			OBJECTIVE To analyze impulsive-like behaviors of SD rats induced by pramipexole in Y-maze avoidance tasks. METHODS Behaviors of SD rats in Y-maze avoidance tasks were recorded with a camera and analyzed by Noldus Etho Vision XT8 software after acute subcutaneous injection of pramipexole(0.1,1 and 10 mg · kg-1),including right reaction numbers of 20 consecutive avoidance tasks,shuttle number of times between the three arms of Y-maze, distance covered in Y-maze and time spent in safe arms during 20 consecutive avoidance tasks. Then,the prepulse inhibition(PPI)of the startle reflex test was used to assess the effect of pramipexole on sensorimotor gating (SG). Effects of pramipexole on the dialyzed content of monoamine neurotransmitter and its metabolites in the striatum and amygdala of SD rats were measured by microdialysis in vivo. RESULTS Compared with normal control group,the rats of pramipexole group showed a significant increase in the shuttle number of times and distance covered in Y-maze between Y-maze avoidance tasks(P<0.01),but a statistically significant decrease in the time spent in safe arms(P<0.01),while the number of right reactions in Y-maze avoidance tasks was not changed. Such premature responses were quite similar to certain impulsive-compulsive behaviors in rodent models,such as five-choice serial reaction time tasks. In the PPI test,pramipexole displayed an impairing effect on SG(P<0.01). The microdialysis results showed that there was an increase of dopamine and 5-hydroxytryptamine in the striatum of pramipexole group, but not statistically significant. Monoamine neurotransmitters and their metabolites were not significantly changed in the amygdala. CONCLUSION Pramipexole can induce impulsive-compulsive behaviors in Y-maze avoidance tasks,which might be attributed to impaired SG.
		                        		
		                        		
		                        		
		                        	
5.Design of a general evaluation scheme to measure hospital patients’satisfaction
Baosheng BIAN ; Shimin FAN ; Shaomei SHANG ; Yan ZHANG ; Wentao GONG ; Weijiao ZHOU ; Xiaoyan JIN ; Huaxing ZHANG ; Huiwen ZENG ; Jing YANG
Chinese Journal of Hospital Administration 2015;(7):500-502
		                        		
		                        			
		                        			Patient satisfaction is an important index of hospital quality evaluation and performance evaluation.This study established multi-attribute patient satisfaction index system based on the theory of customer satisfaction index,determined the index weight by combination weighting approach,and graded the hospitals and adjusted the weights by the RSR method.A set of comprehensive evaluation scheme is initially formed,which is suitable for patient satisfaction evaluation and performance evaluation.
		                        		
		                        		
		                        		
		                        	
6.Observation of the cavum septi pellucidum with MRI
Wentao WANG ; Chuqing ZHAO ; Shaoyong GONG ; Sheng QIU ; Xiangfei CHEN ; Bosen WU ; Kai WANG ; Junping WANG ; Leitao WEN ; Xiaoqun YAO ; Guangfu YANG
Journal of Practical Radiology 2015;(8):1239-1242
		                        		
		                        			
		                        			Objective To study the development characteristics of cavum sepit pellucidun (CSP)in prematures,neonates,infants and adults with MRI.Methods Brain MR images of different subjects including 141 prematures,106 neonates,171 infants and 35 046 adults were observed to determine the incidence and shape of CSP,and to measure its transverse diameter.Results CSP incidences were 100% (141/141)in prematures,97.17% (103/106)in neonates,2.26%(4/177)in infants and 0.82% (287/35 046)in adults respectively,and the CSP was cylinder (44.00%)or triangle in shape (56.00%)in prematures,triangle (76.40%)or fissure in shape (23.60%)in neonates.For infants or adults,each shape accounted for about a third of three kinds of shape respectively.Its mean transverse diameters were 5.7 mm in prematures,4.1 mm in neonates,13.3 mm in infants and 14.3 mm in adults respectivity.Conclusion CSP has different performances at development periods in human being brain.Most close after birth,while fewer remain in the whole life.
		                        		
		                        		
		                        		
		                        	
7.Assessment of risk factors of schistosomiasis transmission control in Jingzhou City
Liangcai HE ; Jiasong WANG ; Xianbing RONG ; Xianhui LU ; Yaosheng ZHAO ; Wentao BIE ; Youxin PENG ; Shaoxiong GONG
Chinese Journal of Schistosomiasis Control 2014;(1):92-93
		                        		
		                        			
		                        			Objective To understand the key risk factors of schistosomiasis transmission in Jingzhou City,so as to provide the evidence for improving the treatment of these risk factors. Methods Each village of six counties was investigated and 3 envi-ronments were surveyed each village for the distribution of Oncomelania snails and animal stools in the field. The results were ana-lyzed and the risk factors of schistosomiasis transmission were assessed. Results The density of living snails was 0.43 snails per 0.1 m2,the frequency of the frames with snails was 9.12%,and no schistosome infected snails were found. All of the animal stools collected from the field were from bovines. The schistosome positive rate of animal stools was 37.50%(3/8)among the environ-ments,and the schistosome infection rate of stools was 8.11%(3/37). The schistosome infection rate of animal stools was 0 near the residence living sites,and the positive rates were 12.50%and 8.33%in the ditches and slopes,respectively(χ2=0.07,P>0.05). Conclusions Bovine is still the main infectious source of schistosomiasis,i.e. the main risk factor of the disease transmis-sion. Therefore,the strategy of controlling bovine should be strengthened.
		                        		
		                        		
		                        		
		                        	
8.Apoptosis of Tim-3~- T cells induced by secretion of galectin-9 in mice
Wentao HE ; Jin YUAN ; Yi XU ; Hongmin ZHOU ; Lanjun CAI ; Hui GUO ; Chao LI ; Liqun ZUO ; Nianqiao GONG ; Zhonghua CHEN
Chinese Journal of Microbiology and Immunology 2010;30(2):144-149
		                        		
		                        			
		                        			Objective To prepare recombinant adenovirus pAd-gal-9 containing murine galectin-9 and explore galectin-9's pro-apoptotic effect on T lymphocytes. Methods The recombinant adenovirus plas-mid pAd/CMV/V5-DEST-gal-9 was prepared by conventional molecular cloning and LR reaction. The pAd/ CMV/V5-DEST-gal-9 linearlized by Pac I was transfected into 293A cells with Lipofectin 2000. Eight days after transfection, the 293A cells were subjected to freeze/thraw circle for three times and the supernatant was collected after centrifugation. Higer titer pAd-gal-9 was produced by large-scale infection of 293A cells with the supernatant containing pAd-gal-9. The supernatant was condensed to get purified pAd-gal-9 by CsCl density gradient centrifugation. After titer determination with gradient dilution of harvested pAd-gal-9 infec-tion in 293A-seeded 96-wells, pAd-gal-9 was used to infect the CHO cell line. Immunohistological assay, Western blot and flow cytometry were employed to ascertain the subcellular location expression of galectin-9. We added solid-phase transgenic CHO cells or freshly-cultured supernatant to medium containing activated T cells to detect the pro-apoptotic effect of galectin-9. Results The pAd-gal-9 was prepared successful. Im-munohistochemical staining of CHO infected with pAd-gal-9 confirmed that galectin-9 was expressed in the cytosol. Intercellular staining indicated that mean fluorescence intensity of galectin-9 was significantly higher in pAd-gal-9-infected CHO group than control group. Supernatant from pAd-gal-9-infected CHO promoted the apoptosis of T cells. The percent of apoptotic T cells was higher than the Tim-3 positive T cells. Conclu-sion CHO infected with pAd-gal-9 can secret galectin-9 to promote the apoptosis of activated T cells via Tim-3-independent mechanisms.
		                        		
		                        		
		                        		
		                        	
9.Ectopic Over-expression of Oncogene Pim-2 Induce Malignant Transformation of Nontumorous Human Liver Cell Line L02.
Ke REN ; Wentao DUAN ; Yujun SHI ; Bo LI ; Zuojin LIU ; Jiangping GONG
Journal of Korean Medical Science 2010;25(7):1017-1023
		                        		
		                        			
		                        			In order to prove that ectopic over-expression of Pim-2 could induce malignant transformation of human liver cell line L02, three groups of cells were set up including human liver cell line L02 (L02), L02 cells transfected with Pim-2 gene (L02/Pim-2) and L02 cells transfected with empty-vector (L02/Vector). Pim-2 expression levels were detected. The morphology, proliferation level, apoptosis rate and migration ability of the cells were detected respectively. Then the cells were subcutaneously inoculated into athymic mice and the microstructures of the neoplasm were observed. Compared with the controls, Pim-2 expression levels were significantly higher in L02/Pim-2 cells (P<0.05), and their morphology had obvious malignant changes. They also showed a significantly increased proliferation rate (P<0.05) and migration capacity (P<0.05), as well as a significantly decreased apoptosis rate (P<0.05). Only the athymic mice inoculated with L02/Pim-2 cells could generate neoplasm, and the morphology of the neoplasm coincided with that of the hepatoma. The results manifest that ectopic Pim-2 gene could be stably expressed in L02/Pim-2 cells. Both the morphological and biological changes of L02/Pim-2 cells demonstrate the trend of malignant transformation. L02/Pim-2 cells could generate hepatoma in athymic mice. In conclusion, Pim-2 could induce malignant transformation of human liver cell line L02.
		                        		
		                        		
		                        		
		                        			Animals
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		                        			Apoptosis
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		                        			Cell Line
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		                        			Cell Movement
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		                        			Cell Proliferation
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		                        			*Cell Transformation, Neoplastic
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		                        			Humans
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		                        			Liver/pathology/physiology
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		                        			*Liver Neoplasms/genetics/pathology
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		                        			Mice
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		                        			Mice, Nude
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		                        			Neoplasm Transplantation
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		                        			*Oncogenes
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		                        			Protein-Serine-Threonine Kinases/genetics/*metabolism
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		                        			Proto-Oncogene Proteins/genetics/*metabolism
		                        			
		                        		
		                        	
10.Effect of N-acetylcysteine on the liver function and mRNA and protein expressions of NF-KB in brain-dead BA-Ma mini pigs
Yan SONG ; Wenzhi GUO ; Xueli GUO ; Gong ZHANG ; Wentao LI ; Shuijun ZHANG
Chinese Journal of Trauma 2009;25(3):275-279
		                        		
		                        			
		                        			Objective To investigate the effects of N-acetylcysteine (NAC) pretreatment on the liver function and mRNA and protein expressions of nuclear factor-KB (NF-KB) in brain-dead BA-Ma mini pigs. Methods The brain-dead model was established by increasing intracranial pressure by a modi-fied, slow and intermittent way. A total of 15 BA-Ma mini pigs were randomly and equally divided into three groups (five in each group), ie, control group (Group C) : treated only with opening and closing abdomen after anesthesia; group without NAC treatment (group B): brain-dead models without use of NAC; NAC treatment group (Group N): 1 and 12 hours after establishment of brain-dead models, 200 mg/kg NAC was added into 100 ml normal saline and intravenously transfused. Levels of ALT and AST in serum as well as TNF-α, IL-1β and IL-6 were determined at 3,6,12, 18,24 hours after brain death. The changes of liver tissues were observed by HE staining under a light microscope, the uhrastruc-rural changes of liver tissues observed under electron microscope, the expression of NF-KB detected by immnohistochemistry and change of NF-KB mRNA by RT-PCR. Results (1) Compared with Group C, serum ALT and AST began to increase at 12 hours after brain death, but IL-1β, IL-6 and TNF-α be-gan to increase three hours after brain death in Groups B and N. mRNA and protein expressions of NF-KB in Groups B and N began to increase six hours after brain death, when Group B increased more sharply than Group N, with statistical difference (P<0.05). (2) At 12 hours after brain death, injury of liver cells in Group B was severer than that in Group N. Conclusion NAC can inhibit the mRNA and pro-tein expressions of NF-KB, decrease the release of inflammatory factors and hence protect the hepatic structure and function during brain death.
		                        		
		                        		
		                        		
		                        	
            
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