~3H-Clonidine, a potent and selective alpha2-adrenergic agonist was used to label alpha2-a-drenoceptors in intact lymphocytes isolated from rat spleen. Binding of ~3H-Clo-nidine was rapid(t1/2: 2min)and readily reversed by 10umol?L~(-1) clonidine(t1/2: 3-4min). ~3H-Clonidine saturationexperiments indicated a single c1ass of site with a K_D of 6.57?1.63nM and Bmax of 72.4?13.4 fmol/10~7 lymphocytes. Adrenergic agonists competed for ~3H-clonidine binding site with anorder of potency:epinephrine)norepinephrine)isoproterenol. These results show the presence ofalpha2-adrenoceptors in splenic lymphocytes. Computer analysis of competition experiments withadrenergic agonists revealed three classes of sites: high affinity site, medium affinity site and lowaffinity site. The affinity of high affinity site is 2-3 orders of magnitude higher than the one ofmedium affinity site, whereas the latter is the same orders higher than low affinity site Using im-proved Mishell-Dutton method, 10umol?L~(-1) clonidine suppressed the Ig M synthesis and thesuppression was blocked by 10 umol?L~(-1) phentolamine. These results indicate the suppression ofIg M antibody response to SRBC in vitro by clonidine is mediated by the alpha-adrenoceptors.

Sufficient embryos are needed for the preservation of Beagle dogs germplasm resources and the prepara-tion of gene?modified human disease animal models. Up to now, the induced ovulation technique has no effect on dogs,it is hard to obtain mature oocytes in vivo, although the scientists try a lot in many aspects, but still could not make a break?through. The in vitro maturation rate is too low to support the preservation of germplasm resources, application in gene?modified disease models and biomedical research. Aiming to provide useful information on breakthrough in dog oocytes mat?uration, this review will summarize the effect of different age and reproductive stage,different morphology and size of the oo?cytes and lipid droplet on the in vitro maturation of dog oocytes.

During medical rescue for nuclear and radiation accidents, triage of potential victims could contribute to better use of medical resources currently and higher efficiency of the rescue.Biological dose estimation techniques ( estimated biodosimetry) are effective for assessing the degree of external radiation injury.The application of estimated biodosimetry to triage is important for effective nuclear accident medical rescue.In this paper, the characteristics of the estimated biodosimetry and its application to triage during medical rescue for nuclear accident and radiation were discussed.

Objective To evaluate the effect of perioperative administration of dexmedetomidine on postoperative ileus after laparocolectomy.Methods Sixty patients scheduled for abdominal surgery were randomly divided into two groups,30 in each group.Group D received dexme-detomidine administeration at a loading dose of 0.6 μg/kg for 10 minutes before induction,followed by an infusion rate of 0.5 μg·kg-1 ·h-1 to 30 min before the end of surgery.The control group re-ceived saline instead of Dex.After the surgery,Group C received intravenous sufentanyl 2 μg/kg, while group D sufentanyl 2 μg/kg combined with Dex 2 μg/kg.Heart rate variability (HRV)were detected before Dex infusion (T0 ),10 minutes after intubation (T1 ), 10 minutes after CO 2 insufflation (T2 ),1 hour after CO 2 insufflation (T3 ),10 minutes after CO 2 desufflation (T4 ),and 10 minutes after extubation(T5 ).The plasma concentrations of epinephrine(E)and norepinephrine (NE)were determined at T0 ,T3 ,T5 ,T7 and T1 0.The recovery of bowel function was evaluated in terms of the first time to fart and intake food.Results Compared with T0 ,HRV of power (TP), high-frequency (HF)power,low-frequency (LF)power and the ratio of LF/HF power were signifi-cantly decreased at T1-T4 in group C and at T1-T5 in group D.The plasma concentrations of E and NE were higher at T3 ,T5 ,T7 and T1 0 in both group C and group D (P <0.05).Compared with T1 ,TP, LF and the ratio of LF/HF were increased at T2-T4 (P <0.05).Compared with group C,TP,LF and the ratio of LF/HF were decreased at T2-T5 ,The plasma concentrations of E and NE were decreased at T3 ,T5 ,T7 and T10 and the time of first flatus was earlier(P <0.05).Conclusion The perioperative ad-ministration of dexmedetomidine during laparocolectomy facilitated the early recovery of bowel func-tion after surgery and decreasede the time of postoperative ileus.

Objective: To investigate the effect of sevolfurane (SEVO) post-conditioning on protein kinase B (AKT)/mammalian target of rapamycin (mTOR) pathway for protecting ischemia/reperfusion (I/R) injury in isolated rat’s heart. Methods: A total of 84 isolated rat’s heart prepared by Langendorff method were randomly divided into 7 groups andn=12 in each group.①Sham group,②I/R group,③SEVO post- conditioning (SPC) group,④NVP-BEZ235 solvent dimenthyl sulfoxide (DMSO) group,⑤Phosphatidyl inositol 3-kinase (PI3K)/mTOR dual inhibitor NVP-BEZ235 (BEZ) group,⑥BEZ+SPC group and⑦SEVO alone group. The hearts received 30 min ischemia followed by 120 min reperfusion with relevant treatment except for Sham group and SEVO group in which the hearts were without ischemia process. Myocardial infarct (MI) size and tissue pathological changes were observed, protein expressions of phosphor-AKT (P-AKT)/total-AKT, P-mTOR/total-mTOR, Beclin1, Bax/Bcl-2 and cleaved Caspase-3 were examined at the end of reperfusion respectively. Results: Compared with I/R group, SPC group presented decreased MI size (26.28±4.00) % vs (49.22±3.66) % and reduced tissue pathological changes; up-regulated protein expressions of P-AKT/total-AKT and P-mTOR/total-mTOR by 79.85% and 67.02%, while down-regulated protein expressions of Beclin1, Bax/Bcl-2 and cleaved Caspase-3 by 33.77%, 69.26% and 48.84%respectively, allP<0.05. Compared with SPC group, BEZ+SPC group sowed increased MI size (53.85±4.06) % vs (26.28±4.00) %and elevated tissue pathological changes; down-regulated proteins expressions of P-AKT/total-AKT and P-mTOR/total-mTOR by 46.06% and 42.95%, while up-regulated protein expressions of Beclin1, Bax/Bcl-2 and cleaved Caspase-3 by 29.90%, 206.85% and 114.65% respectively, allP<0.05. Conclusion: SPC may activate AKT/mTOR pathway and inhibit cardiomyocyte autophagy and apoptosis, thereby attenuate I/R injury in isolated rats’ heart.

Objective To investigate the effects of Saikosaponin A (SSA)on cognitive function and cAMP/CREB signaling pathway and expression of BDNF in mice after traumatic brain injury. Methods Sixty SD male mice were randomized into three groups:shame operation group (group S, n =20),trauma group (group T,n =20)and SSA treatment group (group A,n =20).Mice received an administration of SSA 5 mg/kg (group A)or equal volume saline (group S,group T)immediately and once daily for 5 consecutive days after trauma.The cognitive function was detected by Morris wa-ter maze test on day 1,3,7 and 14 after trauma.The hippocampal tissues were harvested after be-havioral tests and homogenized for measuring the levels of brain derived neurophic factor (BDNF)and cyclic AMP (cAMP)by ELISA as well as the levels of cAMP-response element binding protein (CREB)and phosphorylation-cAMP-response element binding protein (pCREB)by western bolt. Results Compared with group S,the escape latency and swimming distance were significantly pro-longed in group T on day 1,3,7 and 14 and group A on day 1,3 after trauma (P <0.05 );while compared with group T,they were significantly shorter in group A on day 7,14 after trauma (P <0.05).Compared with group S,the levels of BDNF,cAMP,CREB and pCREB were significantly de-creased in group T(P < 0.05 ).Compared with group T,the levels of BDNF,cAMP,CREB and pCREB were significantly increased in group A (P <0.05).Conclusion SSA can significantly improve cognitive dysfunction in mice after traumatic brain injury,and the mechanism may be related to the activation of cAMP/CREB signaling pathway and up-regulation of BDNF.

Objective To investigate the impact of enhanced recovery after surgery (ERAS)on immune function in patients undergoing laparoscopic hepatectomy.Methods Sixty patients undergoing laparoscopic hepatectomy (34 males,26 females,aged 38-57 years,ASA Ⅰ or Ⅱ),were randomly divided into two groups:enhanced recovery after surgery group (group E)and non-enhanced recovery after surgery group (group C).The patients in group E received enhanced recovery after surgery,while the patients in group C received routine perioperative management and anesthesia methods.The operation method and time,the volume of bleeding,the intraoperative fentanyl con-sumption,the volume of fluid input,the preoperative and postoperative CVP and temperature were recorded in the two groups.Blood samples were obtained before induction (T0 ),at the end of opera-tion (T1 ),on day 1 (T2 ),day 3 (T3 ),day 7 (T4 )after operation for determination of plasma con-centration of IgA,IgM,IgG and the percentages of T lymphocyte subsets (CD3+ ,CD4+ ,CD8+ ) and CD4+/CD8+ ratio were detected with flow cytometry.Furthermore,the visual analogue scale (VAS)score and Ramsay score were evaluated 4 hours,8 hours,24 hours and 48 hours after opera-tion in two groups.Results Compared with group C,the intraoperative fentanyl consumption,the volume of fluid input and the postoperative CVP in group E were significantly decreased,while the postoperative temperature was significantly increased (P < 0.05 ).Compared with T0 , the percentages of CD3+ ,CD4+ ,the CD4+/CD8+ ratio and the plasma concentration of IgA,IgM,IgG in group E on T1-T3 were significantly decreased,the percentages of CD3+ ,CD4+ ,the CD4+/CD8+ratio and the plasma concentration of IgA,IgM,IgG in group C on T1-T4 were significantly decreased (P <0.05).Compared with group C,the percentages of CD3+ ,CD4+ ,the CD4+/CD8+ ratio and the plasma concentration of IgA,IgM,IgG in group E on T1-T4 were significantly increased (P <0.05),the visual analogue scale (VAS)score 4 hours,8 hours,24 hours after operation were signifi-cantly decreased (P <0.05).The comparision of Ramsay scores at all the time points between two groups were similar.Conclusion ERAS applied to patients undergoing laparoscopic hepatectomy can reduce the intraoperative fentanyl consumption,prevent the occurrence of hypothermia and provide satisfactory postoperative analgesia,which can significantly improve the immune function in patients.

Objective To investigate the expression and relationship between Phosphatase and tension homolog deleted on chromosome ten (PTEN) and Alpha fetoprotein(AFP) in hepatocellular carcinoma. Methods PTEN and AFP expression was examined by immunohistochemical method. Results PTEN expression was significantly weaker in patients with tumor less than 5.0 cm and patients with CLIP score≥1 (P<0.05). The expression of PTEN and AFP was negatively correlated ( Rs=-0.422, P=0.043), serum AFP and tissue expression of AFP was positively correlated (Rs=0.380, P=0.042), but for individual expression was not consistent. Conclusions PTEN and AFP is respectively negative and positive adjustment factor of PI3K/AKT pathway. Tissue expression of AFP may be more significant, and may become the new targets for the treatment of hepatocellular carcinoma.

Objective To evaluate the effect of stellate ganglion block (SGB) on cellular immune function in diabetic rats.Methods Healthy male Sprague-Dawley rats,aged 3 months,weighing 240-280 g,were used in this study.Diabetes mellitus was induced by intraperitoneal 1％ streptozotocin 60 mg/kg and confirmed by blood glucose ≥ 16.7 mmol/L 3 days later.Forty-eight rats with diabetes mellitus were randomly divided into 2 groups (n=24 each) using a random number table:diabetes mellitus group (group DM) and group SGB.Another 24 healthy rats,aged 3 months,were selected and served as control group (group C).At 1 week after successful establishment of the model,unilateral transection of cervical sympathetic trunk (TCST) was performed in group SGB,while the right cervical sympathetic trunk was only exposed in C and DM groups.Before TCST (T0) and on 1,3,7 days after TCST (T1-3),6 rats were randomly selected from each group,and blood samples were collected from the inferior vena cava for determination of the blood glucose,plasma norepinephrine (NE) concentrations (by enzyme-linked immunosorbent assay),and levels of T lymphocyte subsets CD3+,CD4+ and CD8+ in whole blood (using FACSCalibur flow cytometer).C D4+/CD8+ratio was calculated.The rats were weighed before sacrifice,and the rats were sacrificed to obtain the thymus which was weighed.The thymus index (thymus weight/body weight) was calculated.Results Compared with group C,the blood glucose was significantly increased,and the levels of CD3+ and CD4+ in whole blood,CD4+/CD8+ ratio,and thymus index were significantly decreased at T0-3 (P＜0.05),and no significant change was found in CD8+ levels in DM and SGB groups (P＞0.05),the plasma NE concentrations were significantly decreased at T1-3 in group SGB (P＜0.05),and no significant change was found in plasma NE concentrations in group DM (P＞0.05).Compared with group DM,the blood glucose and plasma NE concentrations were significantly decreased,and the levels of CD3+ and CD4+ in whole blood,CD4+/CD8+ ratio,and thymus index were significantly increased at T1-3 (P＜0.05),and no significant change was found in CD8+ levels in group SGB (P＞0.05).Conclusion SGB can improve the cellular immune function in diabetic rats.

Objective To clone the full-length CDS sequence of LOC 401296 , an undefined gene that we found recently, and to obtain both its expression in eukaryotic cells or E.coli and antiserum to LOC401296 protein.Methods The full-length CDS sequence of LOC401296 was amplified by polymerase chain reaction (PCR).Then we established the expression vectors pET28B-LOC401296 and pCMV-Myc-LOC401296 by cloning the full-length CDS sequence into vector PET28B and vector pCMV-Myc respectively.Isopropyl Thiogalactoside (IPTG) was used to induce LOC401296 expression in E.coli.Furthermore,the protein purified and refolded was used to immunize BALB /c mice.The titer of the antiserum collected from immunized mice was identified by ELISA assay and Western blot .Results We cloned the full-length CDS sequence of LOC401296 was successfully .Protein LOC401296 was expressed as was expected and the mouse anti-human LOC401296 antiserum was obtained .The antiserum titer reaching 1∶64 000 was identified by ELISA .Besides, Western blot analysis showed that the antiserum could be used to detect protein LOC 401296 .Conclusion The full-length sequence of LOC401296 is obtained and the mouse anti-human LOC401296 antiserum becomes available .This study can contribut to further research on the undefined gene LOC 401296 .