Objective To afford the basis for the application of Sinomenine(SIN) in the proposal for organ transplantation, the expression of TGF-? 1 was explored in inborn WistarSK kidney transplantation model. Methods Forty-eight recipients were divided into 4 groups at random:group 1 being used as control and treated with 0 9% natrii chloride(3ml?kg -1 ?d -1 ,ip), group 2 treated with SIN(30mg?kg -1 ?d -1 ,ip),group 3 treated with CsA(2 5mg?kg -1 ?d -1 ,ip),group 4 treated with SIN(30mg?kg -1 ?d -1 ,ip) combined with CsA(2 5mg?kg -1 ?d -1 ,ip).Allografts of 6 recipients from each group were harvested at day 6 before end stage rejection for histological studies. The synergistic effects of SIN combined with CsA were assessed by COX proportional hazards model. The quantitative methods of immunohistochemistry were used for analysis on expression of TGF-? 1.Results Control recipients rejected their allografs at day 9 post-operation, the average life time was 7 83?9 75 days. Treated with either SIN(30mg?kg -1 ?d -1 ,ip) or subtherapeutic doses of CsA(2 5mg?kg -1 ?d -1 ,ip) allografts survival was mininally prolonged. Combined treatment with SIN and subtherapeutic doses of CsA led to a significant prolongation of graft survival(group 4), more than 18 days. The expression of TGF-? 1 was enhanced significantly in group 2 and group 3.No synergistic effect of SIN combined with CsA on expression of TGF-? 1 was observed. Conclusion The therapeutic value of SIN in rat kidney allograft model suggests that this safe chinese medicine has remarkable efficacy of anti-acute rejection in rat kidney transplantation. It shows significant synergistic effect with sub-therapeutic CsA. The immunosuppressive efficacy of SIN is probably based on enhancing express of TGF-? 1. No synergistic effect of SIN and CsA on expression of TGF-? 1 indicates low danger of fibrosis of allografts. SIN will be a hopeful chinese medicine for immunosuppressive treatment in organ transplantation.

Total 118 patients with pyogenic liver abscess admitted to Zhongshan Hospital were classified into pypgenic liver abscesses of cryptogenic origin (PLAC,n =54),pyogenic liver abscesses of biliary origin (PLAB,n =48) and pyogenic liver abscesses of hemagenous origin (PLAH,n =16)according to the causes.The clinical features of three groups were compared and analyzed.The results showed that no statistically significant difference existed among three groups in clinical manifestation and signs.Compared to PLAB,the abscesses in right lobe of liver were more common in PLAC,diabetes mellitus was more prevalent in PLAC,but the co-existing diseases including chronic liver disease,non biliary malignant tumor and ALP elevation were less in PLAC.Klebsiella pneumoniae was frequently detected in PLAC,but there was no significant difference comparing to other two groups.No significant difference was found among three groups in the choice of therapeutic methods,the course of antibiotics,length of hospital stay and the days from admission to diagnosis.

The clinical data of 118 patients with bacterial liver abscess admitted to a tertiary hospital from January 2007 to December 2009 were analyzed retrospectively.Etiology study showed that 45.8％(54/118) cases of bacterial liver abscess were cryptogenic,40.7％ (48/118) were biliary source and 13.5％ (16/118) were hematogenous.The positive rate of pus culture was 57.1％ (36/63),among which klebsiella pneumoniae accounted for 72.2％ (26/36).The positive rate of blood culture was 31.2％(20/64) and klebsiella pneumoniae accounted for 95％ (19/20).One hundred and sixteen patients (98.3％) underwent color ultrasound examination with an accuracy rate of 93.2％.84 patients (71.2％)underwent the CT scan with an accuracy rate of 95.3％ and 20 patients underwent MRI with an accuracy rate of 85％.Seventy five patients (63.6％)accepted ultrasound-guided percutaneous aspiration and catheter drainage combined with antibiotic therapy; 40 patients(33.9％)were treated with antibiotics alone.On the discharge 27 patients(22.9％) achieved clinical cure,87 patients (73.7％) were classified as effective,4 patients (3.4％) had no effect and no death in this group of patients.The average length of hospital stay was 16 days.Cryptogenic was the most common cause of bacterial liver abscess,biliary source took the second place.Kleibsiella pneumoniae was the predominant organism.Detailed medical history inquiry,physical examination and assistant examination may increase the accuracy rate of diagnosis.Needle biopsy should be performed if necessary.Ultrasound-guided percutaneous aspiration and catheter drainage combined with antibiotics should be the first-line treatment.

Objective To investigate the changes of stress response on inhalation-intravenous general anesthesia during pericystectomy for Liver hydatid cyst. Methods Thirty ASA I-II patients with liver hydatidosis for pericystectomy were studied. The blood from right jugular vein were collected for measurement of serum cortisol (Cor) concentrations and glucose (BG) levels, and MAP, HR and BIS were observed and recorded at different point-times: pre-intubation for 1 min, post-intubation for 3 min, pre-incision for 1 min, post-incision for 3 min, post-incision for 10 min, post-surgical exploration 10 min. The data of pre-intubation and pre-incision served as controls. Results The levels of Cor concentration of post-incision for 10 min were decreased than those of pre-intubation and pre-incision (P<0.05); MAP of post-incision for 10 min and post-surgical exploration for 10 min were increased than those of pre-intubation and pre-incision (P < 0.01); For HR, the data of post-surgical exploration 10 min was much quicker compared with pre-incision (P < 0.01), which is slower than that of pre-intubation(P<0.01). Conclusion Inhalation-intravenous general anesthesia may inhibit the stress response during intubation,incision and surgical exploration for liver hydatidosis pericystectomy. Hemodynamic changes during incision may reflect the trend of stress response in advance.

Objective To evaluate the effect of RNA interference in p53 gene on the expressions of genes involved in ultraviolet B (UVB)-induced premature senescence and photocarcinogenesis in human skin fibroblasts (HSFs).Methods A previously established HSF cell clone with repressed expression of p53,which was named as HSF-p53,was cultured and irradiated with a subcytotoxic dose (10 mJ/cm2) of UVB once a day for five consecutive days.The HSFs with normal expression of p53 served as the control.Subsequently,β-galactosidase (SA-β-gal)-staining was performed to estimate the degree of senescence,quantitative real-time PCR array was performed to determine the mRNA expressions of photocarcinogenesis-and senescence-associated genes,including p53,p21,p19,p16,pRb,fibronectin,osteonectin,smooth muscle 22 (SM22),bax,bcl-2,hypoxia-inducible factor-1 α (HIF-1α),vascular endothelial growth factor(VEGF),and human double minute-2 (hdm2).Statistical analysis was carried out by Student's t test using the software SPSS 10.0.Results The percentage of SA-β-gal-positive cells in irradiated HSF-p53 was 19.70％ ± 0.85％,significantly higher than that in unirradiated HSF-p53 (12.77％ ± 0.81％,t =6.45,P ＜ 0.05),but lower than that in irradiated control HSFs (50.48％ ± 5.30％,t =7.86,P ＜ 0.05),and similar to that in unirradiated control HSFs (18.50％ ± 0.45％,t =2.57,P ＞ 0.05).Compared with the control HSFs,the HSF-p53 showed decreased expressions of p21,p19,fibronectin,osteonectin,SM22 and bax genes (all P ＜ 0.05),but increased expressions of bcl-2,HIF-1α,VEGF and hdm2 genes (all P ＜ 0.05),and a similar expression of p16 gene (P ＞ 0.05); the repeated UVB radiation significantly promoted the expressions of p16 and pRb genes (both P ＜ 0.05),but had no obvious effect on the expressions of the other genes in HSF-p53 compared with unirradiated HSF-p53 (all P ＞ 0.05).Conclusions The inhibition of p53 expression may decelerate the UVB-induced premature senescence in HSFs,which may be involved in the p53-dependent tumor suppression.

A cytotoxicity model of primary cultured hepatocytes with D-galactosamine (D-Gal) has been established. The ALT value of culture medium was used as the marker of cell injury. Using this model, the in vitro antihepatotoxic effects of hepatic stimulator substance (HSS), isolated from regenerating adult rat liver, as well as Potenlin and 15-amino acid solution were observed. The results showed that although HSS could stimulate DNA synthesis in serum-free culture of adult rat hepatocytes, it had no effect to reduce ALT value in vitro. However the latter could be reduced by Potenlin or 15-amino acid solution in high concentrations. The described model may be useful for preliminary screening of antihepatotoxic activity of drugs.

Objective To evaluate the effects of different doses of dexmedetomidine on lung injury in pediatric patients undergoing open heart surgery under cardiopulmonary bypass (CPB).Methods Forty-eight pediatric patients,aged 7 months-3 yr,of ASA physical status Ⅱ or Ⅲ,scheduled for elective repair of ventricular septal defect under CPB,were randomly divided into 4 groups (n =12 each) using a random number table:control group (group C) and 3 different doses of dexmedetomidine groups (D1-3 groups).Anesthesia was induced with midazolam,fentanyl and vecuronium.The patients were endotracheally intubated and mechanically ventilated.In D1-3 groups,dexmedetomidine 0.5μg/kg (initial dose) was given over more than 10 min,followed by continuous infusion at 0.1,0.3 and 0.5 μg· kg-1 · h-1,respectively,until the end of operation.The equal volume of normal saline was given instead of dexmedetomidine in group C.Before administration (T1),and at 2,6 and 12 h after termination of CPB (T2-4),blood samples were collected for determination of plasma tumor necrosis factor-α (TNF-α),interleukin-6 (IL-6) and IL-8 concentrations and for blood gas analysis.Respiratory index (RI),oxygenation index (OI) and lung compliance (CL) were calculated.Results The plasma concentrations of TNF-α and IL-8 at T2-4 and IL-6 at T3,4 were significantly lower in D2,3 groups than in group C.Compared with group D1,the plasma concentrations of TNF-α,IL-8 and RI were significantly decreased,and OI and CL were increased at T2-4 in D2-3 groups,and the plasma IL-6 concentrations at T3,4 in group D2 and at T2-4 in group D3 were decreased.The plasma concentrations of TNF-α and IL-6 were significantly lower,while OI and CL were higher at T2,3,and the plasma concentrations of IL-8 and RI were lower at T2-4 in group D3 than in group D2.Conclusion Dexmedetomidine can reduce lung injury in pediatric patients undergoing open heart surgery under CPB and promote the lung function after CPB possibly through inhibiting inflammatory responses to CPB dosedependently.

[ ABSTRACT] AIM:To study the effects of anti-aging Klotho protein on neonatal rat myocardial cells with hypo-xia/reoxygenation ( H/R) injury.METHODS:The cardiomyocytes of neonatal SD rats were cultured to establish hypoxia/reoxygenation model.The myocardial cells were divided into normal control group, H/R model group, different concentra-tions of Klotho protein (0.1μmol/L, 1μmol/L and 10μmol/L) pretreatment groups.The myocardial cells pulse frequen-cy was observed before and after H/R.The cell viability was measured by MTT assay.The leakages of LDH, CK and AST, the content of MDA and the activity of SOD were detected.The apoptotic rate of the myocardial cells was analyzed by flow cytometry.The mRNA expression of endoplasmic reticulum stress markers and apoptosis-related molecules GRP78, CRT, CHOP and caspase-12 was measured by real-time PCR.The protein levels of CHOP, caspase-12 and phosphorylated Akt in the myocardial cells were determined by Western blot.RESULTS: Compared with normal control group, the pulse fre-quency, cell viability rate and SOD activity of myocardial cells were significantly decreased, the cell apoptotic rate as well as the contents of LDH, CK, AST and MDA were increased in H/R model group.The mRNA expressions of GRP78, CRT, CHOP and caspase-12 as well as the protein levels of CHOP and caspase-12 were increased, whereas p-Akt level was decreased obviously.Compared with H/R model group, the pulse frequency, cell viability rate and SOD activity were in-creased significantly, the cell apoptotic rate as well as the contents of LDH, CK, AST and MDA were decreased in Klotho pretreated group.The mRNA expression of GRP78, CRT, CHOP and caspase-12 as well as the protein levels of CHOP and
caspase-12 were decreased, while p-Akt level increased significantly.CONCLUSION:Anti-aging Klotho protein improves the myocardial cell survival and inhibits the apoptosis by increasing the resistance of the cells to oxidative stress and exces-sive endoplasmic reticulum stress response, which is related with the activation of Akt phosphorylation in H/R-injured my-cardial cells.

AIM:To study the protective effect of anti-aging Klotho protein on human umbilical vein endothe-lial cells (HUVECs) treated with high glucose (HG).METHODS:HUVECs were cultured in vitro, and divided into PBS control group, 5.5 mmol/L glucose group, 33.3 mmol/L glucose group, 0.1 μmol/L Klotho +33.3 mmol/L glucose group, 1 μmol/L Klotho+33.3 mmol/L glucose group , and 10μmol/L Klotho+33.3 mmol/L glucose group .The viabili-ty of the HUVECs was measured by MTT assay .The content of malondialdehyde ( MDA) , and the activities of lactate de-hydrogenase (LDH), superoxide dismutase (SOD) and glutathione (GSH) in cell culture supernatants were observed . The production of reactive oxygen species ( ROS) in HUVECs was analyzed by flow cytometry .The levels of nitric oxide ( NO) , endothelin ( ET-1 ) , intercellular adhesion molecule-1 ( ICAM-1 ) in HUVEC culture medium were detected by ELISA.The protein expression of nuclear factor-kappa B (NF-κB) in the HUVECs was determined by Western blot .RE-SULTS:Compared with PBS control group , 33.3 mmol/L glucose significantly decreased the HUVEC viability , increased ROS, LDH and MDA levels , reduced the activities of SOD and GSH , decreased the NO secretion , and induced the ET-1 and ICAM-1 secretion and the protein expression of NF-κB in HUVECs.When HUVECs were treated with Klotho protein at different concentrations combined with 33.3 mmol/L glucose, the cell viability was increased significantly , the ROS, LDH and MDA levels were decreased significantly , the antioxidant SOD and GSH activities were significantly increased , the se-cretion of NO was increased , but ET-1 and ICAM-1 releases and protein expression of NF-κB were significantly reduced . CONCLUSION:Anti-aging Klotho protein promotes the viability of HUVECs treated with HG , reduces the oxidative dam-age and ROS production , and restores the normal secretory function of HUVECs , thus playing a protective role in vascular endothelial cells through reducing the protein expression of NF-κB.

Objective To investigate the effects of different BIS values on postoperative cogni-tive dysfunction (POCD)and S100βprotein(S100β)in the early stage of postoperation.Methods Fifty patients who were scheduled for selective abdominal surgery under general anesthesia (male 34 cases, female 1 6 cases,aged 65 to 75 years,ASA Ⅰ or Ⅱ)were randomly divided into two groups:light anesthesia group (group L,n =25,BIS value was maintained at 50 to 59)and deep anesthesia group (group D,n =25,BIS value was maintained at 30 to 39).BP,HR,SpO 2 ,ECG,PET CO 2 ,inhaled anes-thetic concentration and BIS values were recorded on time points of 5 minutes after the patients ente-ring the operating room (T0 ),before endotracheal (T1 ),intubation (T2 ),incision (T3 ),two hours after incision (T4 ),three hours after incision (T5 )and at the end of surgery (T6 ).The procedure du-ration,anesthesia time,dosages of propofol,fentanyl,midazolam and VAS scores on 1 d after sur-gery were also recorded.Blood samples were collected on time points of 10 min before anesthesia,im-mediately after surgery and 24,48 h after operation.S100β concentration were detected.Mini-mental State Examination (MMSE)score and Trail Making Test (TMT)completion time were recorded on 1 d before surgery and 1,3,7 d after surgery.Results BIS value of group D were lower than group L on T2 ,T3 and T4 .The propofol dosage of group D was significantly greater than that in group L (P <0.05 ).The concentration of serum S100βincreased significantly immediate and 48 h after operation in both groups compared with 10 min before anesthesia(P < 0.05).It was still higher 24 hours after op-eration than before anesthesia.But there was no statistic difference.Compared with the end of surger-y,the concentration of serum S100βin two groups on 24 h after surgery were significantly decreased (P < 0.05 ).The concentration of serum S100β in group L on the end of surgery and 24 h after surgery were higher than that in group D significantly (P <0.05).Compared with 1 d before surgery, postoperative 1 d MMSE scores in two groups and postoperative 3 d MMSE score in group L de-creased significantly (P <0.01).Compared with postoperative 3 d,postoperative 7 d MMSE score in group L increased significantly (P <0.01).Postoperative 1,3 d MMSE score in group D were signifi-cantly higher than group L (P <0.05).Compared with 1 d before surgery,TMT completion time in two groups on 1 d after surgery were significantly prolonged (P <0.01 ).Compared with 1 d after surgery,TMT completion time in two groups on 3 d after surgery were significantly shortened (P <0.01).Compared with 3 d after surgery,TMT completion time in group L on 7 d after surgery was significantly shortened (P <0.01 ).TMT completion time in group D on 1,3 d postoperative were significantly shorter than group L (P <0.05).POCD incidence of group D on 1 d after surgery was lower than that in group L (P < 0.05).Conclusion Different depth of anesthesia can ensure hemo-dynamic balance in old patients during surgery and after surgery.When BIS value was maintained at 30 to 39,it had lower S100βprotein levels,lower incidence of early POCD and a lesser degree of post-operative cognitive dysfunction.