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 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.

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 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.

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 To investigate the relationship between human leptin receptor(LEPR)gene G3057A polymorphism and type 2 diabetes complicated with non-alcoholic fatty liver disease(NAFLD).Methods 216 cases of newly diagnosed type 2 diabetes(104 cases complicated with NAFLD)and 108 cases of normal glucose tolerances(NGT)were recruited.Hemi-nested PCR-RFLP and PCR direct sequence analysis were conducted to detect the polymorphisms of LEPR G3057A polymorphism.The plasma leptin and insulin levels were measured by ELISA kit.Plasma lipid and glucose metabolic parameters were measured routinely.Liver ultrasound scanning Was carried out among all subjects.Results Type 2 diabetic patients complicated with NAFLD had higher plasma alanine aminotransferase(ALT),triglycerides(TG),low density lipaprotein cholesterol(LDL-C),leptin levels and lower plasma insulin levels than those cages without NAFLD and NGT.The variant frequency at nucleotide 3057 G to A transversion Was 76.0% in type 2 diabetic patients complicated with NAFLD,which Was also significant higher than those cases without NAFLD(62.1%)or NGT cases(53.2%)(x2=14.63,P＜0.01).Conclusions The polymorphism of LEPR gene 3057 probably contributes to the onset of NAFLD in type 2 diabetes by regulating lipid metabolism and affecting insulin sensitivity.

The rats were assigned to blank control group, classical induction group, and drynaria total flavonoid group. Whole bone marrow culture method was applied to isolate and purify rats bone mesenchymal stem cells (BMSCs). Akaline phosphatase activity, calcium nodes, TGF-β1 and BMP-2 secretion in the process of bone mesenchymal stem cells osteogenic differentiation were detected. The results showed that compared to the blank group and classical group, drynaria total flavonoid promoted osteogenic differentiation accompanied with increased TGF-β1 and BMP-2 secretion (all P＜0. 05). Drynaria total flavonoid may promote osteogenic differentiation of BMSCs via upregulating TGF-β1 and BMP-2 expressions, and play an active role in the treatment of osteoporosis.