Objective To investigate the effects of ketamine on the renal ischemia-reperfusion (IR) injury in rats and the underlying mechanism. Methods Twenty-four male Wistur rats weighing 220-250 g were randomly divided into 4 groups (n=6 each): sham operation group (group S), IR group, ketumine 2 mg/kg group (group K_1), ketamine 10 mg/kg group (group K_2). The rats were anesthetized with intraperitoneal chloral hydrate 300 mg/kg. Renal ischemia was induced by clamping the left renal artery for 45 min followed by 6 h of repednsion using an atraumatic clamp. In group K_1 and K_2, ketaminc 2 and 10 mg/kg were injected via the caudal vein 5 min before the repedusion respectively. The rats were killed at 6 h of reperfusion, and blood samples were collected from the right auricle for measurement of serum creatiniue (Cr) and blood urea nitrogen (BUN) concentrations. Pathological changes in renal tissues were observed with light and electron microscopes. The expression of Fas and Caspsse-3 in the renal tubular epithelial cell was determined by immuno-histochemistry. The apeptosis in the renal tubular epithelial cell was detected by TUNEL assay. Apeptotic index (AI) was calculated. Results Compared with group S, the levels of serum Cr and BUN, expression of Fas and Caspase-3 and AI were significantly increased in group IR, K_1 and K_2 (P < 0.01). The levels of serum Cr and BUN, expression of Fas and Caspase-3 and AI were significantly lower in group K_(1,2) than in group IR and in group K2 than in group K_1 (P<0.01). The microscopic examination showed that the renal IR injury was obviously attenuated in group K_2 compared with group K_1. Conclusion Ketamine can attenuate the renal injury induced by IR in a dose-dependent manner in rats. The underlying mechanism may be related to the inhibition of apoptosis in the renal tubular epithelial cell.

ObjectiveTo investigate the effects of induction with Dezocine to prevent the agitation of the patient after remifentanil anesthesia.Methods60 patients of upper abdominal surgery,ASA Ⅰ ～ Ⅱ class,were randomly divided into 3 groups,20 patients in each group,anesthesia was induced with the midazolam 0.05 mg/kg,propofol 1 ～2mg/kg,vecuronium 0.12mg/kg.Analgesic drugs were administer as follow:control group,remifentanil ( group Rmf )1 μg/kg; fentanyl group ( group F) 4 μg/kg; Dezocine group ( group D) 0.2mg/kg.Remifentanil、propofol are used to maintain anesthesia.The time of recovery and extubation,the VAS pain score,the Ramsay score and adverse reactions such as vomiting,respiratory depression after extubation were recorded.ResultsCompared with group Rmf,the hemodynamic parameters was more stable in group F and group D.Compared with group F ( 1 μg/kg),the VAS pain score,the Ramsay score and adverse reactions significantly decreased in group D ( P ＜ 0.05 ),and the time of extubation in group F was longer than group D(P ＜ 0.05).Conclusion0.2mg/kg of Dezocine could be used for the induction of anesthesia to reduce the incidence of agitation and adverse reactions during recovery period.

Objective To investigate the effects of cell proliferation and apoptasis on the develop-ment of gastric mucosal lesion in patients with primary pathological duodena-gastric reflux (DGR). Methods Gastroscopy, histologie examination of gastric mucosal biopsy and 24-hour intra-gastric bilirubin monitoring with Bilitec 2000 were performed in 58 patients with primary pathological DGR. Immunohisto-chemical staining was used to detect the expressions of Ki-67 and Bcl-2 proteins. Cell apoptosis in gastric mucesa was determined by TUNEL technique. Results The proliferating index (PI) and apoptosis index (AI) in patients with primary pathological DGR were significantly higher than those in control group (P< 0.05). The differences of PI and AI between high reflux group and low reflux group were significant (P< 0.05). The incidence difference of chronic superficial gastritis (CSG) and chronic atrophic gastritis (CAG) in gastric antrum between the two groups was significant (P<0.05). With lesion progressing from normal gastric mucosa, CSG, CAG to intestinal metaplasia (IM), PI and AI increased gradually and consistently. PI was still on the rise after dysplasia (Dys), but AI decreased. The positive expression rate of Ki-67 in Dys were significantly higher than that of other groups (P<0.05), so was that of Bcl-2 (P<0.05). Conclusion Cell proliferation and apoptesis may be one aspect of the main pathogenesis of gastric mucesa lesion and cell dysplasia in patients with primary pathological DGR. Over-expreasion of Ki-67 and Bcl-2 proteins in CAG, IM and Dys may play a key role in the development of gastric cancer.

Objective To study the association among gastric mucosal lesions caused by primary pathological duodenogastric reflux(DGR),H.pylori infection,and bile reflux.Methods Twenty-four hour intragastric bilirubin monitoring were performed on 58 patients with primary pathological DGR.The patients were divided into high reflux group(n=29)and lOW reflux group(n=29)based on the severity of bile reflux(＜23.60%).The association among gastric mucosal lesions,H.pylori infection,and bile reflux were analyzed.Results The positive rate of H.pylori infection was 20.7% (6/29)in high reflux group and 48.3%(14/29)in low reflux group(P＜0.05).The frequency of intestinal metaplasia in gastric antrum and angularis in high reflux group was higher than that in low reflux group(P＜0.05).The pathological scores of gastric antrum and angularis in H.pylori positive group and high reflux group were higher than those in H.pylori negative group and low reflux group (P＜0.05).The time percentage of bilirubin absorbance≥0.25 in H.pylori positive group was lower than that in negative group(P＜0.05),while the difference in short reflux frequency,long reflux frequency,longest reflux time,maximum,mean and median value of absorbance between H.pylori positive and negative groups showed no significant difference(P＞0.05).The time percentage of bilirubin absorbance≥0.25 was positively correlated with pathological scores of gastric antrum and angularis in both H.pylori positive and negative groups(P＜0.05),but was not correlated with that of gastric body(P＞0.05).Conclusions In patients with primary pathological DGR,excessive bile reflux is related to chronic lesion of gastric mucosa.regardless of H.pylori infection.Bile reflux may inhibit H.pylori to locate in gastric mucosa.H.pylori infection and bile reflux may co-contribute to gastric mucosal lesions.

AIM:To investigate the effects of angiotensin II ( Ang II) on the immune maturation and the oxi-dized low-density lipoprotein (Ox-LDL)-uptaking capacity of human monocyte-derived dendritic cells (DCs).METH-ODS:Human peripheral blood mononuclear cells were isolated by density gradient centrifugation , and the monocytes were purified by positive selection with anti-CD14 magnetic beads.After cultured with rhGM-CSF (100 μg/L) and rhIL-4 (50μg/L) for 5 d, the monocytes differentiated into immature DCs .On the 6th day of the culture, the cells were treated with various concentration levels of Ang II or pretreated with losartan .The immunophenotypic expression of HLA-DR and CD83 was analyzed by flow cytometry .The secretion levels of IL-12 and IFN-γin the culture supernatants were measured by ELISA.Furthermore, DCs were incubated with DiI-labelled Ox-LDL.The DiI-Ox-LDL-incorporated fraction was investiga-ted by flow cytometry .The mRNA expression of 3 scavenger receptors , scavenger receptor A ( SR-A) , CD36 and lectin-like oxidized low-density lipoprotein receptor 1 (LOX-1), was examined by real-time PCR.RESULTS: Ang II induced the maturation of human monocyte-derived DCs, stimulated the expression of CD83 and HLA-DR, and promoted the secre-tion of IL-12 and IFN-γ, which were suppressed by losartan .Furthermore, Ang II increased the Ox-LDL-uptaking capacity of DCs, which was partially reduced by losartan .The incubation of DCs with Ang II enhanced the mRNA expression of LOX-1 in a dose-dependent manner , which was reduced by losartan .However, the expression of SR-A and CD36 was not changed .CONCLUSION:Ang II promotes the immune maturation of human monocyte-derived DCs and increases the up-take of Ox-LDL probably through the up-regulation of LOX-1 expression.

Objective To investigate efficient diagnosis and treatment of 17α-hydroxylase (17OHD) deficiency by summarizing clinical characteristics of those patients.Methods From January 1983 to January 2010,48 cases with 17OHD in Peking Union Medical College Hospital were studied retrospectively.Results Among 48 patients with 17OHD,karyotype analysis showed,12 cases with 46,XX and 36 cases with 46,XY.The 46,XX karyotype and 46,XY karyotype with complete 17OHD had typical clinical presentation of amenorrhea [ 12/12,100％ ( 36/36 ) ],no typical spontaneous puberty [ 12/12,13.9％ (5/36) ],Hypertension [ 11/12,100％ ( 36/36 ) ],hypokalemia [ K +:( 2.6 ± 0.7 ),( 2.8 ± 0.7 )mmol/L],hypergonadotropin [ follicle-stimulatinghormone ( FSH ):( 51 ± 35 ),( 79 ± 46 ) U/L,luteinizing hormone( LH ):( 27 ± 14 ),(49 ± 37 ) U/L ],impaired production of sex hormones [ testosterone(T):0.003,0.005 nmol/L; estradiol ( E2 ):26.86,10.64 pmol/L ],hyper-progesterone [ (P):( 32 ± 15 ),( 29 ± 23) nmol/L],impaired production of 17α-hydroxyprogesterone ( 17α-OHP ) [ ( 2.5 ± 1.1 ),( 2.4 ±1.7) nmol/L],ACTH hypersecreation (91.8,114.0 pmol/L).ACTH stimulating test did not elevated in 17α-OHP and cortisol.Conclusion When patients with elevated basal serum levels of progesterone higher than that of ovulation period in addition to clinical symptoms,examination about 17OHD should be warranted.

Objective To investigate the metabolite changes in the preschool children with autism spectrum disorder (ASD) using MR spectroscopy (MRS) and explore the associations between image findings and clinical variables, which may provide a noninvasive brain biochemical method for the early diagnosis and prevention of autism. Methods Twenty one cases of preschool ASD children (3-6 years old) and age-and sex-matched 20 preschool healthy controls underwent single voxel short (SVS) short TE (TE=30 ms) MRS. The absolute metabolite concentrations in the anterior cingulate cortex (ACC) , anterior middle anterior cingulate cortex (aMCC) and posterior cingulate (PCC) were quantitatively analyzed using LCModel software. Two independent sample t tests were used for analysis. The relationships between metabolite concentrations and diagnostic and statistical manual of mental disorders (DSM-IV) , childhood autism rating scale (CARS) and autism behavior checklist (ABC) were analyzed by Pearson correlation analysis. Results Compared to control subjects, ASD patients had significantly lower N-acetylaspartate (NAA) values (4.35 ± 0.80, 6.34±0.82, 8.04±0.97 mmol/L respectively) in ACC, aMCC and PCC (t=2.640, P=0.012;t=2.182, P=0.035;t=3.343, P=0.002) , had significantly lower choline (Cho) 1.32±0.22 mmol/L (t=2.905, P=0.006) and glutamine and glutamate complex (Glx) 10.02 ± 0.88 mmol/L (t=2.090, P=0.043) in PCC. Cho, total creatine (tCr) , myo-Inositol (MI) and Glx levels did not differ between groups in other aforementioned regions (P>0.05). Negative correlations between the NAA ualues in the PCC and CARS (r=-0.504, P=0.020) were detected, and no significant correlation among DSM-IV, CARS, ABC and other metabolite values (P>0.05). Condnsions The biochemical changes in the preschool children with ASD in cingulate reflect the neuronal loss, structural or functional damage and cell membrane enzyme metabolic dysfunctions, may reveal the pathological basis of ASD. These results may provide noninvasive and quantitative methods for the diagnosis and prognosis evaluation of ASD child.

Aim To study the expression of human VEGF165 cDNA in Pichia pastoris and to obtain high-level expressed recombinant human VEGF165 with good biological activity. Methods Amplifying human VEGF165 cDNA by PCR, after confirmed by DNA sequence analysis, the gene was inserted into the Pichia pastoris expression vector pPIC9K containing AOX1 promoter and α secreting signal peptides, the recombinant expression plasmids pPIC9K/hVEGF165 was constructed and transformed into KM71. The multiple insert transformants were screened， fermented in flasks and induced by 10 mL/L methanol. Results After 4 days of methanol induction, the expressed hVEGF165 reached up to 60％ of total proteins in supernatant by SDS-PAGE. Western blot assay proved the expressed hVEGF165 having good antigenicity and high specificity. The recombinant protein was further purified with Heparin-Sepharose CL6B affinity chromatography, and was proved to have good biological activity in stimulating HUVEC proliferation. Conclusion High-level expression of secreted hVEGF165 has been successfully achieved in Pichia pastoris expression system.

Aim To investigate the effect and mecha-nism of valproic acid on neuroglobin expression, and the neuroprotective role of valproic acid against H2 O2-induced neurotoxicity. Methods Western blot, RT-PCR and luciferase assay were used to detect the pro-tein levels, mRNA levels and promoter activity of mouse and human neuroglobin induced by valproic acid. Luciferase assay was used to investigate the role of transcription factor CREB in the up-regulation of neuroglobin by valproic acid. MTT assay was used to evaluate the effect of valproic acid against H2 O2-in-duced neurotoxicity. Results VPA treatment marked-ly increased the protein levels, mRNA levels and pro-moter activity of Ngb in mouse N2 a cells and human SKNSH cells. CREB specific inhibitor KG501 or CREB dominant negative mutant KCREB attenuated VPA-induced Ngb promoter activity. VPA could pro-tect N2a cells from H2 O2-induced neurotoxicity. Con-clusion CREB mediates VPA-induced Ngb up-regula-tion, which may contribute to the neuroprotective effects of VPA in oxidative stress in neurons.

Purpose:The study was designed to investigate the feasibility of contrast-enhanced ultrasound in the differentiation of benign and malignant adnexal masses.Materials and Methods:Sixty-nine consecutive patients with adnexal masses received trans vaginal contrast-enhanced ultrasound.The image and perfusion features were assessed.Results:All of 26 malignant tumors showed detectable contrast enhancement,including 24 cases with a quick,heterogeneous or branching pattern.Among 39 benign lesions,24 were cystic with circle or half-circle enhancement,including 5 cases with intra-cystic septum or papillae slightly enhanced.The other 15 cases were solid,8 of them had slightly dotted enhancement.There are significant difference in enhancement patterns between benign and malignant masses ( P < 0.0001).The 4 cases of borderline tumors showed progressive,heterogeneous enhancement.Conclusion:Contrast-enhanced ultrasound is of value in the differentiation of benign and malignant adnexal masses.