1.Preparation and identification of mouse anti-human B7-H6 monoclonal antibodies
Chinese Journal of Microbiology and Immunology 2016;36(7):519-522
Objective To prepare mouse anti-human monoclonal antibodies against B7-H6 and to identify their biological characteristics. Methods The B7-H6 gene was cloned by RT-PCR from a human lung adenocarcinoma cell line ( A549 ) and then subcloned into the eukaryote expression vector pCMV3 to construct the recombinant vector pCMV3-B7-H6. The recombinant vector pCMV3-B7-H6 that was verified with enzyme digestion and gene sequencing was transfected into NIH/3T3 cells by electroporation. BALB/c mice were immunized with the successfully transfected cells named 2H8 through intraperitoneal injection. The monoclonal antibodies against human B7-H6 with the advantages of high affinity and specificity were pre-pared by using hybridoma technology. Western blot assay and flow cytometry analysis were used to identify the specificity of prepared monoclonal antibodies. Results The recombinant eukaryotic expression vector encoding B7-H6 was successfully constructed. Two hybridoma clones that stably secreted monoclonal anti-bodies against B7-H6 were screened out by using flow cytometry analysis and the monoclonal antibodies se-creted by them were belonged to IgG2a isotype. Specific reactions between B7-H6 and the secreted mono-clonal antibodies were confirmed by Western blot assay and flow cytometry analysis. Conclusion The mon-oclonal antibodies which recognized B7-H6 specifically were prepared successfully.
2.Effect of Sevoflurane Inhalation Anesthesia on Stress Reaction and Kynurenicacid Level in Patients Under-going Laparoscopic Cholecystectomy
Qingquan HE ; Jianshu CHEN ; Haifeng YANG
China Pharmacist 2016;19(6):1102-1104
Objective:To study the effect of sevoflurane inhalation anesthesia on stress reaction and kynurenic acid ( KYNA) level in serum of the patients with laparoscopic cholecystectomy .Methods:Totally 48 patients undergoing laparoscopic cholecystectomy were randomly divided into two groups.Group S(n=24) received sevoflurane inhalation and group V (n=24) received intravenous general anesthesia.The levels of norepinephrine (NE), epinephrine (E), cortisol (Cor), KYNA and neutrophil and the neutrophil/lympho-cyte ratio were measured at five time points, namely T1(before the anesthesia), T2 (just after the surgery) , T3(in the morning of the first day after the surgery ) , T4 ( in the morning of the second day after the surgery ) and T5 ( in the morning of the third day after the surgery).The correlation of Cor, NE, E and NLR with KYNA was studied as well.Results:The levels of Cor, NE and E at the three time points after the surgery (T2, T3 and T4) in group V were significantly higher than those before the surgery (T1) with statistically significant difference (P<0.05), while those in group S had no significant changes (P>0.05).And the levels of Cor, NE and E at the three time points after the surgery (T2, T3 and T4) in group S were markedly lower than those in group V at the same time point (P<0.05).After the surgery (T2, T3, T4 and T5), KYNA levels in group V were significantly decreased and NLR values in group V were significantly increased when compared with those of T 1.In group S, KYNA levels and NLR values after the surgery (T2, T3, T4 and T5) were markedly increased , which were higher than those in group V at the same time point with statistically significant difference (P<0.05).The correlation analysis indicated the level of KYNA displayed positive correlation with the levels of NLR , Cr, Z and E (P<0.05).Conclusion:Sevoflurane inhalation anesthesia can inhibit the increase of stress reaction and KYNA level in the patients with laparoscopic cholecystectomy , which may be related with its anti-in ammatory properties .
3.Effects of propofol combined with remifentanil on hepatic ischemia-reperfusion injury in cirrhotic rats
Jianzhen WANG ; He YANG ; Lin CHEN
The Journal of Clinical Anesthesiology 2014;(7):709-712
Objective To investigate the effects of propofol combined with remifentanil on he-patic ischemia-reperfusion injury in cirrhotic rats.Methods Sixty male SD rats of 260 to 300 grams were randomly divided into five groups(n=12):the sham-operated group(group S);the model con-trol group (group M);propofol group (group P);remifentanil group (group R);propofol combined with remifentanil group (group PR).In group M,P,R,PR,the hepatic arteries and veins of middle and left lobes were occluded for 20 min after 1 w hepatocirrhosis by using four principal factors,and group S went through an open surgery only.In groups P,R,and PR,porpofol (at a rate 20 mg·kg-1 ·h-1 for 1 h)、remifentanil (at a rate 1 μg·kg-1·min-1 for 1 h)and porpofol (at a rate 20 mg·kg-1· h-1 )combined with remifentanil(at a rate 1 μg·kg-1·min-1 for 1 h)was infused iv at 10 min before is-chemia,respectively.In group M,normal saline was infused iv at the same rate.Blood samples were taken at the end of 4 h reperfusion to determine serum AST,ALT activity.Meanwhile liver specimens were collected to assess Bcl-2 and Bax protein expression in liver cell and measuring the apoptosis in-dex(AI)of hepatic cell.The pathological change of liver was also examined.Results Compared with group S,the activity of AST,ALT,the expression of Bcl-2 and Bax and the apoptosis index(AI)of hepatic cell of the other groups all increased significantly(P <0.05);Compared with group M,the expression of Bcl-2 of groups P,R,PR significantly increased,and the activity of AST,ALT,the ex-pression of Bax and the apoptosis index(AI)of hepatic cell significantly decreased(P <0.05 );Com-pared with groups P and R,the expression of Bcl-2 of group PR significantly increased,and the activ-ity of AST,ALT,the expression of Bax and the apoptosis index(AI)of hepatic cell significantly de-creased(P <0.05);Microscopy and scanning electron microscopy showed that,for groups P,R,PR, pathological injury of liver tissue significantly reduced compared with group M.Conclusion Propofol combined with remifentanil can reduce the hepatic ischemia-reperfusion injury in cirrhotic rat by regu-lating Bcl-2 and Bax protein expression and inhibiting apoptosis.The effect is much more enhanced when they are used in combination than individuals.
4.STUDY ON IMMUNE RESPONSE TO THERAPEUTIC HBV DNA VACCINE
Guangming CHEN ; Fuqiang YANG ; Xiaoqian HE
Medical Journal of Chinese People's Liberation Army 2001;0(12):-
In this study,we constructed 2 eukaryotic expressing plasmids namely pcS2?S and pFP,encoding HBV preS2+S envelope protein and human IL 2/IFN ? fusion protein, respectively. The double plasmid fusion protein was used as an adjuvant in preparation of a treatment type HBV gene vaccine. To investigate its feasibility for use as a therapeutic DNA vaccine, we've evaluated the immune response after injection into healthy mice, HBV transgenic(Tg) mice, New Zealand rabbits and Rhesus monkeys. The results showed that the therapeutic DNA vaccine can improve: (1)the CTL activity; (2)the HBsAg(30?g/ml) specific T lymphocyte proliferation in which the stimulation index(SI) and cytokines(IL 2/IFN r) release levels of the pS2.S immunized group(SI=5.6?0.9; 226.3?41.1/51.1?7.1pg/ml) were significantly higher( P
5.Changes of organ water content in the early stage of 50% third degree burns in dogs
Baubin HE ; Zongzheng YANG ; Faming CHEN
Journal of Third Military Medical University 1984;0(02):-
In order to verify the effects of postburn fluid infusion on the tissue water content,extravascular water volume,residual blood water volume,and total water volume of the major organs were measured in dogs inflicted with 50% TBSA third degree burns and the dogs received no fluid infusion,immediate infusion or delayed infusion respectively.It was found that the water contents of most of the canine organs,no matter they received fluid infusion or not,were higher than those of the control.The increase was especially marked in the extravascular water volume and less marked in the residual blood water volume,which suggests that the increase of water content of most of the organs of the burned animals results mainly from tissue edema rather than congestion and/or stasis of blood.
6.CELLULAR IMMUNE RESPONSE INDUCED BY DNA VACCINE IN BOTH HEALTHY AND HBV TRANSGENIC MICE
Fuqiang YANG ; Guangming CHEN ; Xiaoqian HE
Medical Journal of Chinese People's Liberation Army 1981;0(06):-
The peptides from HBV cytotoxic T lymphocyte(CTL) epitope were used to either stimulate or impact the immune effector (E) or CTL target (T) cells respectively, in order to investigate the cellular immune response induced by DNA vaccination in both healthy and HBV transgenic (Tg) mice. It was found that HBV DNA based immunization could induce CTL activity in healthy BALB/c mice, with intensity correlated with the E/T ratio and IFN ? secretion level in its supernatants. The target cells hit by HBsAg CTL epitope peptide(pp20) could be lysed by the active CTL induced by HBV DNA vaccine encoding preS2 and HBsAg, while the cell lysis could not be observed in the target cells impacted by the HBcAg CTL epitope peptide (pp10). The supernatant IL 12 secretion level (211 3?39 8pg?ml -1 ) in the DNA vaccination group was significantly( P
7.The effects of using the nursing intervention during the course of cure patients with depression after the stroke
Feng YANG ; Xiangmei CHEN ; Xichun HE
Chinese Journal of Practical Nursing 2006;0(17):-
Objective To study the effects of using the nursing intervention during the course of cure patients with depression after the stroke. Methods Divided 127 patients with depression after stroke into the treatment group(63 cases) and the control group(64 cases) randomly,the general nursing intervention and the traditional nursing measures were adopted respectively in the two groups.Compared the effects of treatment between the two groups by the questionnaires. Results Results The effects of treatment in the treatment group were significant better than those of in the control group. Conclusion The general nursing intervention can effective improve the depression of patients with stroke,and then ameliorate their quality of life,shorten the time of recovery.
8.Effects of paclitaxel(poly-L-Iactide/polyglycolic acid)degradable materials on growth of smooth muscle cells
Yu CHEN ; Shuquan LI ; Min HE ; Xiaonan HE ; Haishan YANG
Chinese Journal of Tissue Engineering Research 2009;13(21):4037-4040
BACKGROUND: The problem of the high rate of acute restenosis at an early and advanced stage has been a hot spot, while the stent with paclitaxel (poly-L-lactide, PLLA/polyglycolic acid, PGA) degradable material will resolve it. OBJECTIVE: To seek an ideal biologic degradable material used in biologic degradable stent. DESIGN, TIME AND SETTING: The comparative cytological study was performed at the Laboratory of Toxicology, Institute of Public Health, Jilin University (Key Laboratory of Radiobiology of Ministry of Public Health of China, Key Laboratory of Toxicology of Jilin Province) from July 2003 to July 2005. MATERIALS: Paclitaxel degradable material (PLLA/PGA) (PLLA:PGA= 9: l ) were offered by Changchun Applied Chemistry Institute of Chinese Academy of Science). Human umbilical arterial smooth muscle cells (SMCs) were purchased from Boster, Wuhan, China. METHODS: The sixth passage of SMCs were digested by 0.25% Trypsin under ambient temperature for 3 minutes, incubated in 10 mL 10% ox serum BMEM, and made into SMC suspension. The materials kept in hypothermal disinfectant Co60 were heated to 37 ℃ by waterbath after surface treatment, and then placed in a 6-well culture plate. SMC suspension was added into the middle of the materials. The density of cells was about 5×106/cm2. Cell suspension diffused around the materials gradually. At last, 10% ox serum BMEM culture solution was added into it, and cultivated in a 5% CO2 incubator at 37 ℃. The growth of cells in and surrounding the materials was observed by inverted microscope everyday. MAIN OUCOME MEASURES: The materials were obtained after culture for 3, 6, 12, 19, 26, 34 days and vacuum dehydration, and were weighed. The weight loss of materials was compared before and after culture. The average degradable rate of materials was calculated.RESULTS: Degradable material had no influence on the development of SMCs. The average degradation rate ex vivo was 0.45% per day. CONCLUSION: PLLA/PGA with good cellular compatibility could be applied to intravascular stents.
9.The impacts of low-dose corticosteroids infusion given in different manners on refractory septic shock ;patients
Zhi CHEN ; Chunli YANG ; Huiwei HE ; Zhaohui HE
Chinese Critical Care Medicine 2015;(6):443-447
Objective To discuss the influence of different ways of low-dose corticosteroids infusion on hemodynamics, changes in blood glucose level and prognosis in patients with refractory septic shock. Methods A prospective single-blind randomized controlled trial was conducted. Refractory septic shock patients admitted to the Department of Critical Care Medicine of Jiangxi Provincial People's Hospital from April 1st, 2013 to October 31st, 2014 were enrolled for the study. The patients were divided into control group and research group by random number table. Besides conventional treatment for septic shock, patients in control group were given 200 mg/d hydrocortisone intravenous infusion lasting for 2 hours, while those of research group were given 8.33 mg/h hydrocortisone per hour with an intravenous pump. Treatment lasted for 5 continuous days for both groups. The changes in heart rate ( HR ), mean arterial pressure ( MAP ), central venous pressure ( CVP ) and arterial blood lactic acid in both groups were observed at the time of enroldment and 6 hours, 24 hours, 48 hours, and 5 days after the treatment. With a dynamic blood glucose monitor, mean blood glucose ( MBG ) level, largest amplitude of glycemic excursions ( LAGE ), glucose variability ( GV ), and the ratio of hyperglycaemia time were recorded. The duration of shock, length of intensive care unit ( ICU ) stay, total length of hospital stay, and 28-day mortality of both groups were recorded. Results Seventy-nine septic shock patients were assigned to the treatment, with 41 in control group, and 38 in research group. Compared with control group, 6-hour MAP in research group was obviously lowered [ mmHg ( 1 mmHg=0.133 kPa ):66.31±4.38 vs. 68.58±4.86, t=1.062, P=0.033 ], but there were no significant differences in HR, MAP, CVP, lactic acid clearance and norepinephrine ( NE ) utilization rates at other time points between two groups. No significant difference in MBG was found between research group and control group ( mmol/L:8.69±2.14 vs. 9.95±3.87, t=1.771, P=0.080 ), but LAGE, GV, the ratio of hyperglycemia time in research group were significantly lower than those of the control group [ LAGE ( mmol/L ): 17.18±8.97 vs. 22.71±11.80, t = 2.331, P = 0.022; GV ( mmol/L ): 2.57±1.05 vs. 3.16±1.37, t=2.136, P=0.036;the ratio of hyperglycemia time:( 43.1±11.7 )%vs. ( 49.4±15.3 )%, t=2.044, P=0.044 ]. There was no statistical difference in the following features between research group and control group, such as the duration of shock ( days:3.47±0.98 vs. 3.61±1.07, t=0.605, P=0.547 ), length of ICU stay ( days:8.74±3.12 vs. 9.97±3.37, t = 1.543, P = 0.120 ), total length of hospital stay ( days: 18.34±9.27 vs. 19.58±9.83, t = 0.576, P = 0.566 ) and 28-day mortality rate ( 23.68%vs. 26.83%,χ2=0.103, P=0.748 ). Conclusions Compared with slow intravenous infusion, a continuous intravenous supplementation of small amount of hydrocortisone to patients with refractory septic shock could stabilize blood glucose levels and maintain metabolic balance efficiently. However, in both groups there was no significant difference in the efficiency in stabilizing hemodynamics, shortening shock duration, reducing ICU or hospital days and decreasing 28-day mortality.
10.Surgical treatment and perioperative management of carotid body tumors
Zhiyong CHEN ; Mingchang HE ; Aiguo YANG ; Ning HE
Chinese Journal of General Surgery 1997;0(06):-
Objective To explore the surgical treatment and perioperative management of carotid body tumors.Methods Fifteen patients with carotid body tumor were operated upon after appropriate perioperative management.Results Among the 15 cases,6 underwent simple tumor excision,7 had co-resection of carotid body tumor and carotid artery,and 2 cases had arterial anastomosis of common carotid artery and internal carotid artery after co-resection of carotid body tumor.There was no mortality after operation in this series.All patients were followed up from 1 year to 3 years (average,15.3months).Operative incisions healed by primary intention.The pressure symptoms of carotid body tumor disappeared after operation,but two patients had temporary headache for 3months,and one patient died of cerebral infarction.Injury of hypoglossal nerve occurred in one patient but it recovered after treatment.There were no recurrences of carotid body tumor after operation.Conclusions Adequate perioperative management can improve the effect and safety of operation,and decrease operative complications of patients with carotid body tumor.