1.Retrospect and prospect of the translational medicine research of severe heatstroke
Medical Journal of Chinese People's Liberation Army 2017;42(2):103-108
There exist a series of problems in heat stroke treatment,such as,pathogenesis is still unclear,clinical classification is too simple and has no intrinsic relation with pathophysiological process and prognosis,missing of indexes for hierarchical diagnosis and prognosis prediction,and lack of targeted therapeutic norms.All of these factors could lead to high mortality and disability by heat stroke.Our research team started an epidemiological investigation of heat stroke since 2002.On the basis of discovering organ injury rule,system info and treatment technology on critical medicine were applied to heat stroke treatment.Research on organ injury mechanism for heatstroke was carried out based on translational medicine idea,and periodic research results were also achieved.A series of key technologies for heat stroke treatment were obtained.These technologies were popularized in 30 hospitals across the country,thus improving ability of heat stroke treatment.
2.The effect and safety of thoracic drainage by central venous catheter in critical patients with pleural effusion
Yi CHEN ; Huasheng TONG ; Lei SU
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care 2013;(4):234-236
Objective To investigate the effect and safety of thoracic drainage by central venous catheter in critical patients with pleural effusion. Methods A prospective study was carried out,in which 46 critical patients with pleural effusion admitted to department of critical care medicine were included,and according to the types of their primary diseases to make matched pairs,and they were divided into two groups:conventional drainage control group and central venous catheter drainage observation group(each 23 cases). The drainage effect and complications were compared between the two groups,acute physiology and chronic health evaluation Ⅱ(APACHEⅡ)score, quantitative pleural effusion and the counts of white blood cells(WBC)and of platelets(PLT),the levels of prothrombin time(PT),procalcitonin(PCT)and C-reactive protein(CRP),etc. were measured on the baseline and 24 hours after the operation. Results There were no significant differences in the APACHEⅡ score,quantitative pleural effusion,amount and duration of drainage,as well as the levels of WBC,PLT,PT and PCT between group control and observation(all P>0.05). The post-drainage CRP(μg/L)level in group observation was much lower than that in group control(77.26±67.20 vs. 106.13±66.23,P<0.01). There was a trend that the incidence of complication in group control was higher than that in group observation(26.1%vs. 4.3%),but the difference was not statistically significant(P>0.05). Conclusions In the comparison between the conventional drainage and thoracic drainage with central venous catheter,the therapeutic effects for treatment of critical patients with pleural effusion were similar, but the stress was milder and the incidence of complications was lower in the latter group. Therefore,the thoracic drainage with central venous catheter is a safe,effective and alternative method to substitute the conventional drainage for critical patients with pleural effusion.
3.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.
4.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
5.The effect of inhibition of dUTPase expression by RNAi on adhesive ability of SW620 cell
Xiaorong LAI ; Huasheng TONG ; Jinbao WU
Medical Journal of Chinese People's Liberation Army 2001;0(07):-
Objective To evaluate the curative effect of valsartan associated with low-dose amiodarone on the recurrence of atrial fibrillation (AF), the left atrial diameter (LAD), P wave dispersion (Pd) and the maximum P wave duration (Pmax) in patients with paroxysmal AF. Methods 76 patients with paroxysmal atrial fibrillation (PAF) were randomized to valsartan (test group) and placebo (placebo group), both associated with low-dose amiodarone, and were followed up for 18 months. The patients were asked to report any episode of symptomatic atrial fibrillation and to perform an ECG as early as possible. AF load, Pmax, Pd and LAD were measured before and at the 6th, 12th, and 18th month after the treatment. Results At least one ECG-documented episode of AF was reported in 16% of the patients in test group and in 41% in placebo group, the difference was significant (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.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
8.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.
9.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.
10.Role and mechanism of endoplasmic reticulum stress and Ca2+ overload in pulmonary endothelial cell damage induced by heat stress
Baojun YU ; Na PENG ; Zhengtao GU ; Huasheng TONG ; Lei SU
Medical Journal of Chinese People's Liberation Army 2017;42(6):488-494
Objective To observe the effect of different temperatures on endoplasmic reticulum stress, calcium overload, mitochondria and cell damage in pulmonary microvascular endothelial cells (PMVEC) induced by heat stress, and clarify the mechanism of endothelial cell injury in the process of heat stress to provide experimental basis for clinical prevention and treatment of heat stree. Methods Heat stress model of PMVEC cell was set up. Control group cells were incubated at 37℃, 5%CO2, while heat stress group cells were incubated at 39℃, 41℃, 43℃ for 2h, respectively, then further incubated at 37℃, 5%CO2 for 6h. Pretreatment of cells with 20μmol/L BAPTA-AM or 50μmol/L CsA before heat stress at 43℃. The protein levels of p-PERK, PERK p-eIF2a, eIF2a, ATF4 and GRP78 were analyzed by Western blotting. Intracellular Ca2+, mitochondrial membrane potential and the changes in mitochondrial permeability transition pore were investigated by flow cytometry. The change of caspase-3 was detected by Caspase Assay Kit. Millicell-ERS Volt-Ohm Meter and Accessories was used for determining the changes of transepithelium electrical resistance (TER). Results Compared with the control group, with the increase of heat stress temperature (41-43℃), the phosphorylation of p-PERK and p-eIF2a protein and the expressions of ATF4 and GRP78 proteins were gradually activated, intracellular Ca2+ increased, MPTP pore was opened, mitochondrial membrane potential decreased, cell permeability increased and apoptosis occurred, and it was the most obvious in the 43℃ heat stress group, and the difference was statistically significant (P<0.05). Pretreatment with Ca2+ inhibitors promoted the recovery of the MPTP hole, mitochondrial membrane potential and cell permeability, and reduced the occurrence of apoptosis. While pretreatment with the mitochondrial protective agent did not reduce the release of Ca2+, but it could promote the recovery of cell permeability and reduce the occurrence of apoptosis. Conclusion Heat stress activates endoplasmic reticulum stress response, induces intracellular Ca2+ overload mediated cell and mitochondrial damages in PMVEC cells, which may be one of the important mechanisms of endothelial cell injury induced by heat stress.