Objective To investigate the effect of ketamine on the expression of HSP 70 in myocardium in severely burned rats for its possible mechanism of myocardial protection. Methods Seventy-two male Wistar rats were randomly divided into 3 groups: normal control group (group C, n=8), burn injury group (group BI, n= 32) and ketamine group (group K,n=32). 30% Wtal body surface area of Ⅲ degree burn model was developed in group BI and group K. Ketamine 20 mg/kg was injected IM in group K 15 min after the burn model was made. Equal volume of normal saline was given instead of ketamine in group BI. Group C received no treatment. The rats were sacrificed at 3, 6, 12 and 24 h after administration in group BI and group K respectively(8 rats at each time point). Myocardial samples were obtained for determination of the expression of HSP 70 by Western blot analysis. The myocardial ultrastructure was observed at 3 h after administration with electron microscope. Results The myocardial damage was milder in group K than in group BI. The expression of HSP 70 was significantly higher at 3, 6, 12 and 24 h after administration in group K and group BI than in group C(P<0.05).The HSP 70 expression was significantly higher at 3 and 6 h after administration in group K than in group BI ( P<0.05). Conclusion Ketamine can reduce the myocardial injury induced by severe burn through up-regulating the expression of HSP 70 in cardiocytes.

Objective To investigate the expression of recombinant IL-37b protein and removal of the endotoxin, and identify its biological activity.Methods The prokaryotic expression vector pET28/IL-37b was constructed and to transform Escherichia coli (E.coli) Rosetta.After induction with IPTG, the recombinant protein was purified through Ni2+-NTA gel column and identified by SDS-PAGE and Coomassie brilliant blue staining.Then, the endotoxin protein was removed and was treated with LPS-stimulated RAW 264.7 cells.The culture supernatant was collected.The expression of IL-6 was detected by ELISA and the biological activity of the protein was identified.Results The recombinant IL-37b with high purity was expressed and the endotoxin produced by prokaryotic expression was reduced, and it was identified to have good biological activity.Conclusions In this study a recombinant IL-37b protein with high biological activity is successfully obtained.

Objective To evaluate the effect of controlled hypotension at the beginning of reperfusion on ischemia-reperfusion (I/R) injury of the liver in patients undergoing hepatectomy.Methods Forty ASA Ⅱ or Ⅲ patients (aged 30-60 years and weighing 40-70 kg) undergoing elective partial hepatectomy for liver cancer were randomly divided into two groups (n =20 each):normal blood pressure group (control group,group C) and controlled hypotension group (group H).In group C,normal blood pressure was maintained during reperfusion,while in group H,controlled hypotension (the mean arterial blood pressure (MAP) was maintained at 60-70 mm Hg) was performed for 10 minutes since the beginning of reperfusion.Hepatic portal was occluded during operation.Venous blood samples were taken before hepatic ischemia (T0,baseline) and after 15 minutes of ischemia (T1) and after 25 minutes of reperfusion (T2) for determination of plasma levels of endothelin (ET),nitric oxide (NO),tumor necrosis factor-alpha (TNF-α) and interleukin-1 (IL-1).Results I/R of the liver led to significant increases in plasma levels of ET,TNF-α and IL-1 and a decrease in plasma level of NO at T1,2 as compared with the baseline values at T0 in both groups.Plasma levels of ET,TNF-α and IL-1 were significantly lower while plasma level of NO was significantly higher at T2 in group H than in group C.Conclusion Controlled hypotension for 10 minutes in the initial stage of reperfusion can attenuate I/R-induced injury to the liver in patients undergoing hepatectomy through balancing ET with NO and inhibiting inflammation responses.

Objective To observe the effect of Zhibo Dihuang Pill(ZDP) on leptin-induced idiopathic central precocious puberty(ICPP) in mice.Methods Eighteen BALB/c female mice aged 18 months were randomized into 6 groups: normal group,model group,high-and low-dose ZDP groups(at the dose of 101.4 and 50.7 g?kg-1?d-1 respectively),and megestrol acetate(3.9 mg?kg-1?d-1)group and gonadotropin releasing hormone analogues(GnRH-A) group(at the first dose of 1573.5 ?g/kg,the second and the third dose of 1180 ?g/kg,once every other day).Except the normal group,the mice in other groups received intraperitoneal injection of leptin 20mg/kg to establish ICPP model.After treatment,the vaginal opening,body weight,uterus weight and ovaries weight as well as the serum levels of luteinizing hormone(LH),estrogen(E2) were detected.Meanwhile,the uterus index and ovaries index were also examined.Results High-dose ZDP had an inhibition on the advance of vaginal opening induced by leptin after modeling for 4~6 days(P0.05).Conclusion The therapeutic effect of ZDP for ICPP is probably related with the inhibition of the advance of hypothalamic-pituitary-gonad axis activation.

Inteleukin-37 is a member of IL-1 family and originally named as IL-1F7.It has five different subtypes (IL-37a-e).It has been reported that IL-37 suppresses pro-inflammatory cytokines production of a variety of immune cells, and further regulate innate immune response. In addition, IL-37 has protective effects on colitis, arthritis, pancreatitis and other inflammatory diseases that induced by exogenous stimuli. In a word, IL-37 is a novel anti-inflammatory cytokine and plays an important role in a variety of inflammation-related diseases.

Objective To establish a C57BL/6 mouse model of intestinal infection induced by S.Typhimurium.Methods In order to improve the infectious sensitivity of S.Typhimurium, C57BL/6 mice were intragastrically given 5% (w/v) NaHCO3.Then mice were challenged with S.Typhimurium.The health condition, survival and body weight of mice were observed from day 0 to day 7 after the bacterial infection.The pathological changes were also examined.Results the mice challenged with S.Typhimurium showed decreased body weight and typical clinical signs, including in appetence, piloerection and low survival rate.Macroscopic dissection revealed that intestinal hyperemia and swelling were founded in the mice challenged with S.Typhimurium.Histopathology showed intestinal epithelial and mucosal damages.Conclusions We have successfully established a C57BL/6 mouse model of S.Typhimurium infection.This model may be of crucial significance for studying the biological functions of associated immunological molecules or cytokines in the process of inflammatory bowel disease induced by S.Typhimurium.

Objective To evaluate the effect of controlled hypotension at the beginning of reperfusion on ischemia-reperfusion (I/R) injury of the liver in patients undergoing hepatectomy. Methods Forty ASA Ⅱ or Ⅲ patients aged 30-60 yr weighing 40-70 kg undergoing elective partial hepatectomy for liver cancer were randomly divided into 2 groups ( n = 20 each): group C normal BP and group H controlled hypotension. Hepatic portal was occluded during operation. In group C normal BP was maintained during reperfusion while in group H controlled hypotension (MAP was maintained at 60-70 mm Hg) was performed for 10 min since the beginning of reperfusion.Venous blood samples were taken before hepatic ischemia (T0 ,baseline) and at 15 min of ischemia (T1) and 25 min of reperfnsion (T2 ) for determination of plasma endothelin (ET), nitric oxide(NO), TNF-α and IL-1 concentrations. Results I/R of the liver led to significant increase in plasma ET, TNF-α and IL-1 concentrations and decrease in plasma NO concentration at T1,2 as compared with the baseline values at T0 in both groups. Plasma ET,TNF-α and IL- 1 concentrations were significantly lower while plasma NO concentration was significantly higher at T2 in group H than in group C. Conclusion Ten minutes controlled hypotension in the initial stage of reperfusion can attenuate I/R-induced injury to the liver in patients undergoing hepatectomy by balancing ET with NO and inhibiting inflammation response.

Objective To investigate the effects of ulinastatin on the expression of Toll-like receptor 4 (TLR4) during myocardial ischemia/reperfusion (I/R) in rats and the possible mechanism.Methods Thirty adult Sprague-Dawley rats,weighing 250-280 g,were randomly divided into 3 groups (n =10 each):sham operation group (group S),I/R group and ulinastatin group (group U).Myocardial ischemia was induced by 30 min occlusion of left anterior descending coronary artery followed by reperfusion.Ulinastatin 1 × 104 IU/kg was injected intravenously at 5 min before reperfusion in group U,while the equal volume of normal saline was given in groups S and I/R.The animals were sacrificed at 5 h of reperfusion and myocardial specimens were obtained for microscopic examination and determination of myocardial apoptosis (using TUNEL),TLR4 expression (by immuno-histochemistry) and TNF-α content (by ELIAS).The apoptotic index was calculated.Results Compared with S group,the apoptotic index,TLR4 expression and TNF-α content were significantly increased in groups I/R and U (P ＜0.05).Compared with I/R group,the apoptotic index,TLR4 expression and TNF-α content were significantly decreased (P ＜ 0.05),and the pathological changes were significantly reduced in group U.Conclusion The mechanism by which ulinastatin alleviates myocardial I/R injury is related to inhibition of TLR4 expression and reduction of inflammatory responses during early I/R in rats.

Objective To investigate the effect of Smad3 on cell migration of A549 and HeLa cells.Methods Primers for pCMV-Myc-Smad3 plasmid construction and siRNA targeting Smad 3 were designed and synthesized .pCMV-Myc-Smad3 plasmid was constructed with molecular cloning techniques .Overexpression of Smad 3 with Myc-tag or silencing of endogenous Smad3, and then scratch assay was used to detect the migration ability of A 549 and HeLa cells in vitro. Results pCMV-Myc-Smad3 plasmid was successfully constructed .Overexpression of Smad 3 significantly up-regulated the migration rate of A549 and HeLa cells.Conversely, in the same cells, silencing of endogenous Smad3 or treatment with Smad3 inhibitor, SIS3, down-regulated the migration rate .Conclusions Smad3 promotes cell migration of A549 and HeLa cells.

Objective To investigate the effect of inhalation of enflurane on the expression of N-methyl-D-aspartate receptor subunit 2B (NR2B) in the hippocampus of the offsprings of rats.Methods Thirty SD rats pregnancy 8-10 day weighing 200-250 g were randomly divided into 3 groups ( n =10 each):control group (Group C),4 h inhalation of enflurane group ( group E1 ) and 8 h inhalation of enflurane group ( group E2 ).Group E1 and E2inhaled 1.7 ％ enflurane (in O2 2 L/min) for 4 and 8 h respectively,while group C inhaled oxygen 2 L/rin for 8 h.The learning and memory functions of the offsprings were assessed at 20 and 30 days after birth by Morris maze test.The expression of the NR2B mRNA were examined by RT-PCR,NR2B protein were examined by mmunohistochemistry.Results Compared with group C,the escape latency was significantly prolonged,the frequency of crossing the original platform was significantly decreased,the staying time at the original platform quadrant was significantly shortened at 3-5 days after the test in group E1 and E2 (.P ＜ 0.05 ),the expression levels of NR2B mRNA and protein were significantly decreased at 20 and 30 days after birth in group E1 and E2 ( P ＜ 0.05).There were no significant differences in the indexes mentioned above between groups E1 and F2 ( P ＞ 0.05).Conclusion Inhalation enflurane in the early pregnancy can result in cognition dysfunction through inhibiting NR2B expression in the hippocampus of the offsprings of the rats.