One alkaline protease from Actinomucor elegans AS3.2778 was purified protein. The enzyme was purified using ammonium sulfate precipitation, ion exchange chromatography, hydrophobic chromatography and size exclusion chromatography method, and its properties were also investigated. The molecular weight of this enzyme is 32 kDa with SDS-PAGE method, optimum temperature is 60℃, optimum pH is 8.5 to 10.5, it is stable in the pH range of 6.0 to 9.0 at < 40℃ temperature, and being completely inhibited by the serine protease inhibitor, PMSF, indicated that it belongs to the serine protease family. Specificity test indicated this protease has extensive selectivity to peptide bones, especially to peptide bones composed of Leucine residue.

Objective To investigate the relationship between the secondary hypertension and pituitary microadenoma,and to explore the diagnostic value of magnetic resonance imaging(MRI) examination for pituitary microadenoma.Methods We analyzed the clinical information,laboratory examination results and imaging data in 21 hypertension patients complicated with pituitary microadenoma.Results The manifestations of the 21 patients were as follows:(1) The onset of hypertension was in young age,complicated with headache;(2) The results of 24-hour ambulatory blood pressure monitor(24h ABPM) showed the disappearance of circadian rhythm of blood pressure,and obvious increase of systolic/diastolic blood pressure;(3) The antihypertensive effect of antihypertensive drugs was not satisfactory;(4) Of adenohypophyseal hormones,adrenocorticotrophic hormone(ACTH) level in most patients and prolactin(PRL) level in a few patients were higher than the normal level,and the other hormones levels were normal;(5) The results of MRI examination presented adrenal cortical hyperplasia,adrenal adenoma and chromaffin tumor in some patients.(6) The results of MRI examination showed pituitary microadenoma in all of the patients.Conclusion For those middle-aged and young hypertension patients on whom antihypertensive effect of drugs is poor,MRI examination for adrenal gland and pituitary gland should be taken as the routine examination.MRI examination is the optimal imaging method for pituitary microadenoma,and supply evidence for the syndrome differentiation and treatment of pituitary microadenoma.

ObjectiveTo investigate the effect of arsenic trioxide (ATO) on anti-dsDNA antibody and the expression of DNA methyltransferase 1-mRNA and CD11a-mRNA in lupus MRL/lpr mice.Methods MRL/lpr mice were divided into three groups:the ATO group,the sodium chloride(NS) group,and the cyclophosphamide(CTX) group.The control group consisted of 20 syngeneic normal C57/BL mice,which were subdivided into the ATO group and the NS group.After two-month treatment,all mice were sacrificed.Blood routine test was conducted by SYSMEX KX21.The anti-dsDNA antibody in the serum were detected by ELISA.The expression of DNMT1 and CD11a was determined by RT-PCR.ANOVA and paired t test were used for statistical analysis.Results① The serum level of anti-dsDNA antibody(0.89±0.07) and the gray scale value of CD11a-mRNA(0.43±0.25) in the ATO group were much lower than those in the NS group of MRL/lpr mice(1.77±0.28,P＜0.01; 0.99±0.31,P＜0.05),while the gray scale value of DNMTI-mRNA (0.32±0.30) was significantly higher than that in the NS group(0.16±0.26,P＜0.05 ).② The serum levels of anti-dsDNA antibody was low in both the ATO group and the CTX group (0.90±0.07,0.66±0.22),and it was higher in the ATO group than that in the CTX group (P＜0.05).The gray scale value of DNMT1 mRNA in the ATO group (0.32±0.30) was higher than that in the CTX group (0.16±0.18,P＜0.05) in MRL/lpr mice,and the gray scale value of CD11a mRNA in the ATO group(0.43±0.25) was lower than that in the CTX group (0.86±0.31,P＜0.05) in MRL/lpr mice.③ There was no difference in above parameters between the ATO-group and NS group in C57/BL mice(P＞0.05).ConclusionArsenic trioxide can reduce the serum level of auto-antibody and reverse low methylation.But it has no effect on normal mice.

Objective To investigate the feasibility and efficacy of vaginal sacrospinous ligament fixation (VSSLF) procedure which reserving uterus in the treatment of uterine prolapse. Methods 8 cases with uterine pro-lapse underwent VSSLF procedures which reserving uterus from July 2006 to December 2008. Those with incontinence d urine simultaneously underwent ventrofixation of the middle piece of the urethra and folding of the ligamentum pos-terius of the uretha and those with anterior or posterior vaginal wall prolapse underwent eolporrhaphia anterior-posteri-or. Results All patients successfully accomplished by VSSLF,without intraoperative or postoperative hemorrhage or organ injury. All patients' prolapse symptoms were relieved efficently according to the pelvic organs prolapse quantifi-cation(POP-Q). Patients were followed-up for 18 months after operation. 8 patients all recovered well without any complication related to the procedure after operation. Conclusion VSSLF is a simple,safe,effective and micro-inva-sire way for patients with uterine prolapse who want to reserve uterus.

Objective To investigate the influence of PPARγ excitomotor RSG and ATRA on gastric cancer SGC7901 cell line proliferation in vitro and its potential mechanism study.Methods Human gastric cancer SGC7901 cell line was cultured in vitro.Experiment samples were divided to blank group,10μmol/L ATRA group, 12.5μmol/L RSG group, 25μmol/L RSG group, 10μmol/L ATRA + 25μmol/L RSG group.Proliferation inhibitory effect was determined by MTI assay.Flow cytometry was used to detect cell cycle, H.E stain was used to observed micrography alteration.Expression of PPARγ protein in gastric cancer cells were measured by immunohistochemistry.PPARγ mRNA in gastric cancer cells were measured by RT-PCR.Results ATAR at concentration 10μmol/L, RSG at 12.5 μmol/L and RSG at 25 μmol/L could inhibit the proliferation of SGC7901 cells in a dose-and time-dependent, and when both agents were combined for 72h, growth inhibition ratio was (29.73 ± 0.69) %.Flow cytometry analysis revealed a cell cycle arrest at G1 and S phase, and when both agents combined, S% was (12.87 ± 0.35 )%, cell micrography tended to be normal when both agents combined.Up-regulation of PPARγ protein and PPARγ mRNA expressions were also observed, those effects were enhanced when both agents combined, and grey scale ratio was 0.646.Conclusion The ATRA and RSG could significantly induced growth inhibition of human gastric cancer SGC7901 cell, which may be associated with cell cycle arrest and inducing differentiation, activation of PPARγ protein and PPARγ mRNA expression.Synergistic effect could be caused by the combined use of the two agents.

OBJECTIVE To study the therapeutic effect and the underlying mechanism of asiatico?side on bleomycin-induced rat interstitial pulmonary fibrosis(IPF). METHODS Male Sprague-Dawley (SD)rats were divided into normal control group,bleomycin 5 mg·kg-1 model group and asiaticoside 50 mg · kg-1 group. The model and asiaticoside group were administrated with bleomycin 5 mg · kg-1 to induce IPF,while the asiaticoside group was administrated with asiaticoside 50 mg·kg-1 by gastric perfusion. Hematein eosin(HE)and Masson staining were carried out to analyze the histopathological changes in the lung. Lung homogenates were used to examine hydroxyproline(HYP) content,and serum samples were used to measure the concentration of interferon-γ(IFN-γ),interleukin-4(IL-4) and tumor necrosis factor-α(TNF-α). In addition,immunohistochemical methods were used to locate lung transforming growth factor-β1(TGF-β1)and adenosine 2A receptor(A2AR)expression,and Western blotting was used to examine the expression levels of TGF-β1 and A2AR. RESULTS On the 7th,14th and 28th days,the scores of pulmonary inflammation were higher in model group than in control group (P<0.01),and the asiaticoside group showed mitigated alveolitis(P<0.01,P<0.05) compared with model group. Compared with control group,the scores of pulmonary fibrosis in model group were elevated(P<0.01),and the asiaticoside group showed reduced pulmonary fibrosis(P<0.05). On the 14th and 28th days,HYP content in the model group〔1.85±0.10,(2.48±0.18)mg·g-1〕was higher than in the control group〔0.79 ± 0.07,(0.84 ± 0.08)mg · g-1〕(P<0.01),but HYP content in the asiaticoside group〔1.32±0.131,(1.71±0.13)mg·g-1〕was lower than in the model group(P<0.05). IL-4 and TNF-αin the asiaticoside group were lower than in model group(P<0.05),but were higher in the model group than in the control group(P<0.01,P<0.05). The expression level of TGF-β1 protein in the asiaticoside group was lower than in the model group(P<0.05),but was higher in the model group than in the control group(P<0.05). The expression level of A2AR protein in the asiaticoside group was higher than in the model group(P<0.05),but was lower in the model group than in the control group(P<0.05). CONCLUSION Asiaticoside can mitigate bleomycin-induced IPF by inhibiting the expression of IL-4, TNF-αand TGF-β1,and raising the level of A2AR.

目的观察心理康复对焦虑、抑郁症状的疗效。方法对212例具有失眠、慢性头痛、头晕或多系统症状而不能用某一种疾病解释的患者,进行焦虑抑郁量表检测,并随机分为康复组(88例)和对照组(124例)。前者给予心理康复治疗,后者给予药物治疗,比较两者的疗效。结果康复组总有效率90.91%,高于对照组(74.19%)(P<0.05)。结论对综合医院中具有失眠、头痛、头晕或多系统症状而不能用某一种疾病解释的患者,应进行焦虑抑郁量表检测并给予心理康复治疗。

To investigate the protective effect of hirudin on kidneys of diabetic rats and its molecular mechanism.

Objective Using CT perfusion (CTP) technique,to investigate the graft perfusion changes in patients with hepatic artery stenosis (HAS) with or without ischemic-type biliary lesions (ITBL) after orthotopic liver transplantation (OLT).Methods Thirteen recipients with HAS received CTP scan of the liver,including 8 with ITBL and 5 without ITBL.For all patients,the diagnosis of HAS was made by CTA,and the diagnosis of ITBL by percutaneous transhepatic cholangiography.CT perfusion indices were obtained,including hepatic artery perfusion (HAP),portal vein perfusion (PVP),total liver perfusion (TLP) and hepatic perfusion index (HPI).Results Of the 13 patients with HAS,mean HAP in patients with and without ITBL was 59.8 and 35.1 ml·min-1 ·100 ml-1 (P =0.021,two-tailed paired Student t test) ; mean PVP was 125.4 and 166.2 ml·min-1·100 m1-1 (P =0.016) ; mean TLP was 185.2 and 201.3 ml· min-1 · 100 ml-1 (P =0.306) ; and mean HPI was 33.6 and 18.2 (P =0.005),respectively.Conclusion Using CTP technique,liver perfusion changes were reflected by measuring CTP indices noninvasively.Compared to those without ITBL in this study,HAP and HPI in patients with ITBL were higher and PVP was lower,which may be contributed to biliary inflammation.

Objective To formulate a five-level pediatric emergency triage standard and evaluate the efficacy and reliability of it in determining severity of emergency pediatric patients.Methods According to the assessment methods in Pediatric Advanced Life Support recommended by American Heart Association and American Academy of Pediatrics,we formulated a five-level pediatric emergency triage standard based on the situation of our hospital and analyzed the data one year before and after the application of it.Results Before and after the application of the triage standard,the average satisfaction rate of emergency patients were (81.28 ± 3.97) ％ and (94.13 ± 4.62) ％,and there was significant difference (P ＜ 0.01) ; the proportion of whom became worse during waiting time were 1.83％ (628/34275) and 0.04％ (16/36 187),and there was significant difference (P ＜0.01) ;the average waiting time of emergency admission patients were (12.71 ± 2.32) min and (3.34 ± 1.95) min,and there was significant difference (P ＜0.01) ;the misjudgment rate of severity were 3.78％ (1 296/34 275) and 0.57％ (205/36 187),and there was significant difference (P ＜ 0.05).Conclusion The five-level triage standard is objective,easy to master,and suitable for pediatric triage.It can quickly sort out critical cases from emergency pediatric patients,which can improve the effectiveness of emergency service,make use of medical resources rationally and somewhat solve the problem of overcrowding.