1.Preliminary analysis of differentially expressed protein profiles in two primary colonic tumor cell lines
Huasheng TONG ; Yali ZHANG ; Bo JIANG
Medical Journal of Chinese People's Liberation Army 2001;0(10):-
Objective To analyze the differences in protein pro fi le between laterally spreading tumor (LST) cell line and SW480 cell line using t wo-dimensional electrophoresis (2-DE). Methods 2-DE was empl oyed to isolate the total proteins of the two cell lines.The gels were stained with silver, and the differential protein expressions were analyzed with Melanin e 3 software. Results The protein spots of the two cell lines w ere 1285?51(LST) and 1184?47(SW480) respectively,containing 96?7 differential protein spots with 50?6 expressed or obviously increased only in LST cell line and 47?5 in SW480 cell line. Conclusion The protein profiles between LST and SW480 cell lins are different and further analysis on the differ ent interested spots is required to acquire the biology-associated proteins spe cific LST.
2.The influence on adhesion between laterally spreading tumor cell and collagen Ⅰ mediated by membrane galectin-1 monoclonal antibody inhibition
Huasheng TONG ; Yali ZHANG ; Bo JIANG
Medical Journal of Chinese People's Liberation Army 1982;0(03):-
Objective To observe the influence on adhesion between LST-R1 cell and collagen Ⅰ mediated by galectin-1 antibody inhibition. Methods Laser confocal scanning microscopy was employed to detect the expression of galectin-1 on LST-R1 cell membrane.LST-R1 cells inhibited by galectin-1 antibody or not were seeded in the 48-well plate coated with collagen Ⅰ (20?g/well), and the morphology of the adhesive LST-R1 cells was observed and the adhesive cells were counted. Results Laser confocal scanning microscopy showed the expression of galectin-1 on the membrane of LST-R1 cells.The binding rate between galectin-1 and its antibody was 38.2%?0.92%. In the galectin-1 antibody inhibiting group,more and more irregular angular adhesive cells appeared, and most of the adhesive cells were growing in singles, while the adhesive cells,mainly clustered, in the control group were round or roudish. The numbers of adhesive LST-R1 cells in the antibody inhibiting group and the control group were 40 4 and 30 4 respectively (P
3.Analysis of the characteristics of endoscopic and histopathological features of colorectal serrated adenomas
Wanwen ZHANG ; Baojun REN ; Huasheng TONG ; Yali ZHANG ; Bo JIANG
Chinese Journal of Digestive Endoscopy 2001;0(02):-
Objective To explore the endoscopic and histopathological features of serrated adenomas (SAs).Methods The data of patients with colorectal polyps diagnosed in the Digestive Endoscopy Center at Nanfang Hospital from January 2002 to July 2005 were reviewed and the detection rate, endoscopic appearances, pit patterns and histopathological features of SAs were analyzed.Results In 1928(16.21%) out of 11 894 patients undergoing colonscopy 2811 polyps were found.Among them 61 patients with 71 polyps were found,with a detection rate of 0.51%.The SAs,larger than hyperplastic polyps obviously,were found in patients 39.44% with diameter (larger than 1 cm).The incidence of pedunculated polyps in SAs (26.76%) was higher than that in hyperplastic polyps(13.25%),but less than in adenomatous polyps (43.95%).The pit patterns of SAs, were typeⅢ pit pattern (41.67%) and type Ⅳ pit pattern (18.33%), this result was similar to adenomatous polyps.The incidences of moderate and severe dysplasia of SAs were higher than those of tubular adenomas but lower than villous adenomas.The canceration rate of SA was 2.82%.Conclusion The endoscopic appearances,these of pit patterns and histopathological features of SAs,were different from hyperplastic polyps essentially, but similar to neoplastic polyps with potential malignancy,which should be emphasized in clinical practice.
4.Construction of RNAi recombinant vector of galectin-1 and establishment of LST-R1 cell lines stably transfected by the recombinant vector
Huasheng TONG ; Yali ZHANG ; Bo JIANG ; Lei SU
Medical Journal of Chinese People's Liberation Army 1982;0(03):-
Objective To construct the RNA interference (RNAi) eukaryotic expression vectors of galectin-1, and establish the LST-R1 cell lines stably transfected by RNAi vectors. Methods Two pairs of small interfering RNAs (siRNA) targeting to galectin-1 mRNA (GenBank: NM002305) were designed. The corresponding single-strand short hairpin interfering RNAs (shRNA), containing BamH Ⅰ, Hind Ⅲ sites and a 9nt hairpin structure, was synthesized and annealed. The annealed product and the linear eukaryotic expression plasmid pSuperior-puro, which were digested with Bgl Ⅱ and Hind Ⅲ, were ligated by T4 ligase to set up the interfering system. The recombined plasmid was identified with EcoR Ⅰand Hind Ⅲ enzyme digestion and sequencing, and co-transfected to LST-R1 cells with pcDNA6/TR with Lipofectamine2000. Positive clones were selected with 0.8?g/ml puromycin. After incubated with 4?g/ml tetracyclin for 48 hours, RT-PCR, Western blotting and immunochemistry were employed to determine galectin-1 mRNA and protein levels. Results Sequencing results suggested that the nucleotide sequence and read frame of RNAi eukaryotic exprssion vector of galectin-1, p-shRNA1 and p-shRNA2 were perfect. Stably transfected LST-R1 cell lines of p-shRNA1-LST and p-shRNA2-LST were established. The relative values of galectin-1 expression in LST-R1 cells, p-shRNA1-LST cells, p-shRNA2-LST cells and p-LST cells by RT-PCR were 0.616, 0.298, 0.373 and 0.641, respectively, and 1.00, 0.07, 0.38 and 0.97 by Western blotting. The p-shRNA1 gave the best interfering effect, which was in conformity with the results of immunochemistry measurement. Conclusions RNAi eukaryotic expression vector of galectin-1 mRNA has been successfully constructed. Establishment of the stably transfected LST-R1 cell lines may lay a foundation to explore the roles of galectin-1 in laterally spreading tumor.
5.Impact of galectin-1 expression on LoVo cell apoptosis
Huasheng TONG ; Yali ZHANG ; Bo JIANG ; Lei SU
Chinese Journal of General Surgery 1997;0(04):-
Objective To observe the changes of LoVo cell growth by galectin-1.Methods Eukaryotic expression vector of galectin-1 was constructed and transfected into LoVo cells using lipofectamineRM 2000.Immunochemistry was employed to detect galectin-1 expresson.LoVo cell proliferation and apoptosis were observed.Results Galectin-1 eukaryotic expression vector pEGFP-C1/GAL1 was successfully constructed.Three cell clones,p-GAL1-LoVo and p-LoVo,transfected with pEGFP-C1/GAL1 and pEGFP-C1 correspondingly,and LoVo were cultured successfully.Galectin-1 expression downregulated Bcl-2 level only occurring in p-GAL1-LoVo cells.p-GAL1-LoVo cell proliferation was similar to p-LoVo and LoVo cells,but p-GAL1-LoVo cell apoptosis was increased(9.61?0.56)%,as compared with p-LoVo and LoVo cells,[(3.56?0.53)% and(3.46?0.46)% respectively](P
6.The study of Ulinastatin to reduce pulmonary inflammation and oxidative injury in rats with severe heatstroke
Yi CHEN ; Huasheng TONG ; Xingqin ZHANG ; Dongxin JIANG ; Lei SU
Chinese Journal of Emergency Medicine 2014;23(8):866-870
Objective To investigate the effect of ulinastatin on acute lung injury in rats with severe heatstroke.Methods Fourty-eight rats were randomly (random number) assigned into control group (HS group,n =12),low dose Ulinastatin group (LUTI group,n =12),high dose Ulinastatin group (HUTI group,n =12) and non-thermal group (Sham group,n =12).Rats were prepared with pre-warm chamber to initiate heatstroke.The change of rectum temperature (Tc),heat-rate (HR) and mean arterial pressure (MAP) under heat-stress were recorded.The time-point of heatstroke onset and Tc >42 ℃ was observed.Arterial blood samples were draw at 0 min,20 min,40 min and 60 min for testing partial pressure of oxygen (PaO2) and partial pressure of carbon dioxide (PaCO2).Bronchoalveolar lavage fluid (BALF) were collected at 60 min,and the concentrations of tumor necrosis factor-α (TNF-α),interleukin-1β (IL-l β) and interleukin-6 (IL-6) in BALF were measured with enzyme linked immunosorbent assay kit.Lung tissues were harvested for observing pathological change and measuring the expression of iNOS with Western blot test.Results Compared with HS group,the time-point of Tc > 42℃ (P =0.00),severe heat-stroke (P =0.00) and the median of survival time (P =0.00) in LUTI and HUTI groups were significantly increased.At 60min after heat-stress,the level of PaO2 in HS group was much lower than those in other groups (P =0.00).But there were no differences between LUTI and HUTI groups (P =0.91).The value of PaCO2 in HS group was much higher than those in other groups (P =0.00).And the differences between LUTI and HUTI groups were no significant (P =0.79).The concentrations of TNF-α,IL-1β and IL-6 in HS group was the highest in four groups (P =0.00,P =0.04 and P =0.04),followed by LUTI,HUTI and Sham group.The concentrations of proinflammatory cytokine in LUTI were higher than those in HUTI group (P =0.02,P =0.00,P =0.00).Compared with HS group,the pathological injuries were alleviated in LUTI and HUTI group (P =0.00).The expression of iNOS in lung tissue of HS group was strengthened than LUTI and HUTI group (P =0.00),and there was a significant difference between LUTI and HUTI group (P =0.03).Conclusion Ulinastatin improves respiratory dysfunction and the prognosis of severe heatstroke rats through reducing the inflammatory and oxidative injury in lung tissue.
7.Reversal effect of FG020318 on multidrug resistance in retinoblastoma cell line SO-Rb50
Huasheng YANG ; Guoying LIU ; Juanjuan XU ; Ping ZHANG ; Danping HUANG
Chinese Journal of Pathophysiology 2000;0(10):-
AIM: To investigate the multidrug resistance(MDR),reversal activity of 2-[4-(2-pyridin-2-yl-vinyl) henyl]-4,5-bis-(4-N,N-diethylaminophenyl)-1(H)-imidazole(FG020318) in a retinoblastoma subline SO-Rb50/VCR,which is resistant to vincristine.METHODS: The procedure of stepwise increase in drug concentrations was used to obtain SO-Rb50/VCR,which was resistant to 200 ?g/L vincristine. The chemosensitivity of this drug resistant cell line with and without FG020318 or cyclosporine A(CSA) were detected by MTT assay and the function of p-glycoprotein(P-gp) was examined by rhodamine 123 accumulation detected with flow cytometry(FCM).RESULTS: FG020318(2.5 ?mol/L) significantly reduced IC50 and increased the rhodamine accumulation in a concentration-dependent manner. It was much stronger than the positive control CSA in reversal of MDR.CONCLUSION: A new tumor MDR modulator FG020318 partly reverses MDR in SO-Rb50/VCR.It may be a promising new drug to tackling MDR.
8.Effect of community-based intervention on the improving of early detection of individuals with high risk nasopharyngeal carcinoma
Hong GUO ; Xiangjun FU ; Huasheng LIU ; Senping LIU ; Jie XIAO ; Ping XIAO ; Shenhua ZHANG ; Xiangdong ZHAO
Chinese Journal of Primary Medicine and Pharmacy 2016;23(8):1121-1125
Objective To analyze the effects of community-based intervention on improving early detection of individuals with high risk nasopharyngeal carcinoma and to explore the interventional mechanism of prevention and treatment in nasopharyngeal carcinoma.Methods Total of 139 344 residents of the Second Chinese Medicine Hospi-tal of Guangdong Province Hospital Community were detected per year.Individuals with high risk nasopharyngeal car-cinoma or nasopharyngeal carcinoma patients were divided into intervention group(69 616 cases) and control group (69 728 cases) according to the random number table.The two groups were given conventional treatment (radiothera-py,chemotherapy and surgery treatment) after diagnosed.The intervention group was interfered with long -term prevention,health care,medical care,rehabilitation,health education and psychological intervention by full-time phy-sician,while the control group was only treated by general education intervention.The awareness of early symptoms and the risk factor of nasopharyngeal carcinoma were analyzed.Additionally,the behavior,knowledge and attitude on nasopharyngeal carcinoma were detected before and after the intervention.Results Community-based intervention significantly improved the rate of early detection of nasopharyngeal carcinoma high-risk individuals ( 75.31% vs. 44.37%,χ2 =5.32,P<0.01) .The awareness rates of major symptoms and the damage of nasopharyngeal carcinoma were 89.34%and 91.78%in the intervention group.In the control group,the rates were 42.17%and 28.47%.The differences of the two groups were significant(P<0.05).Additionally,the awareness of diagnosis standard and good habits were improved significantly in the intervention group compared with the control group(P<0.01).Our results showed that intervention changed the attitude and understanding of patients with nasopharyngeal carcinoma and improved the quality of life of patients with nasopharyngeal carcinoma.Conclusion Intervention increased the early diagnostic rate of nasopharyngeal carcinoma and promoted the quality of life of patients with nasopharyngeal carcinoma.
9.The diagnostic value of Sonoclot in the diagnosis of heat stoke complicated with DIC and heat stroke
Peng WAN ; Huasheng TONG ; Xingqin ZHANG ; Pengkai DUAN ; Youqing TANG ; Lei SU
The Journal of Practical Medicine 2014;(16):2562-2565
Objective To evaluate diagnostic value of Sonoclot in the diagnosis of heat stroke complicated with disseminated intravascular coagulation (DIC). Methods 43 patients with heat stroke and coagulation dysfunction were retrospectively included and divided into 2 groups according to scores by International Society of Thrombosis and Hemostasis (ISTH): DIC dominant group (22 case) and DIC non-dominant group (21case). Regular coagulation tests , routine blood test , D-dimer and Sonoclot tests were performed at admission and their clinical data were compared. Logistic regression analysis was applied to evaluate the relationships between DIC occurrence and Sonoclot parameters. ROC curves were used to evaluate diagnostic value of Sonoclot for the patients with DIC and heat stroke. Results There were no differences in age, sex, central temperature and total hospital stay between the 2 groups except ICU stay, DIC symptoms, outcome, ISTH scores and APACHEⅡscores (P <0.05). ACT and CR correlated with the occurrence of DIC (P < 0.05). The AUC of ACT and ACT combined with CR were 0.854 and 0.877 respectively. The specificity of ACT in predicting DIC was 69.2%with the sensitivity of 90.3%. When combined with CR, both the specificity and the sensitivity were increased to 80.2% and 93.5%, respectively. Conclusions Sonoclot can predict DIC quickly and is effective in the diagnosis of heat stroke patients with DIC.
10.Evaluation of Sterile Effect of Arthroscope Surgical Instruments Using Low-temperature Plasma Sterilizer
Juanli ZHU ; Mingxue ZHANG ; Yubin XING ; Ying NIE ; Huasheng WANG ; Feng SONG ; Li HE ; Lin CHEN ; Zheng ZHANG ; Yujie LIU
Chinese Journal of Nosocomiology 2006;0(05):-
OBJECTIVE To evaluate the feasibility and sterile effect of low-temperature plasma sterilizer produced by Chinese Academy of Sciences for the sterilization of arthroscope surgical instruments.METHODS To explore the cleaning,package,arrangement and sterile effect monitoring of instruments in the process of arthroscope sterilization using low-temperature plasma sterilizer.RESULTS In the sterilization process of arthroscope surgical instruments using low-temperature plasma sterilizer,the chemical indicators and indicating patch were discolored,the monitoring of bio-box and the sterilization monitoring of surgical instruments were all negative.CONCLUSIONS Using the low-temperature plasma sterilizer machine for the sterilization of endoscope surgical instruments can ensure the security of sterilization,make less injury of precise instruments,shorten the sterilization time,accelerate the surgical turnover,enhance the frequency of surgical instruments application,and increase the efficiency.