BACKGROUND: Cathepsin E, an intracellular asparUc proteinase of pepsin family, Is composed of 2 homologous domains, each containing a consensus DTG motif, in which the role of aspartic acid (Asp98) is still unknown. OBJECTIVE: To determine the role of Asp98 in the catalysis of Cathepsin E. DESIGN, TIME AND SETTING: An observation experiment was performed in the Guangdong Pharmacological College between November 2007 and December 2008.MATERIALS: Rat Cathepsin E antibody was prepared according to the references, and the purified with affinity chromatography. PcDNA3.0 was offered from Guangzhou Ganewindows Biotech Ltd. Human embryo kidney 293 cell strain was offered from Stem Cell Bank,Chinese Academy of Sciences.METHODS: The cDNA containing full-length rat Cathepsln E was inserted into pBluescdpt IISK plasmid between Smal and Xhol. Asp98 was replaced with an acidic amino acid termed glutamic acid to produce a D98E mutant by Kunkel gene site-directad mutagenesis and expressed in human embryo renal cells.MAIN OUTCOME MEASURES: Expression of wild type and mutant protein was determined, the wild type and mutant Cathepsin E was purified, the enzyme activity, folding and self-precessing were analyzed. RESULTS: Hetaroganous expression of wild type and mutant rat Cathepsin E were found in human embryo kidney 293 cells, and both were expressed in the cell culture media and cell lysate. Mutant D98E was still able to auto-activate into mature enzyme, although its activity was only 13% of that of wild type. The Km value of mutant was similar to that of wild type, but its K<,cat> value was significantly smaller than that of wild type. Both mutant and wild type were inhibited by pepstatin and Ascaris pepsin inhibitor with similar Ki values. The CD spectra of mutant and wild type were similar.CONCLUSION: The Asp98 plays an important role in catalysis, and it can be partly replaced by glutamic acid. Mutant D98E has a lowered activity since the mutation affects the catalytic efficiency.

Objective To examine whether diabetes mellitus increases the risk of pneumonia mortality among seniors in the U.S. general popula-tion. Methods&Results The NHANES III follow-up study data were used. After excluding individuals from other minorities, being hos-pitalized with pneumonia in the previous year at baseline, or death of pneumonia during the first year of follow-up, a total of 3,707 subjects aged 65 years or older (1,794 men and 1,913 women) who had no missing information on variables for the analysis were included. Approxi-mately 16% of seniors at baseline were diabetics, which was defined as either having been diagnosed by a physician, currently taking pills/insulin lowering blood glucose, or HbA1c higher than 6.4%. During an average 11 years of follow-up, a total of 98 deaths due to pneu-monia were recorded (ICD-10:J12-J18). Cox-regression models were used to estimate the risk association between pneumonia mortality and diabetes mellitus. After adjustment for the covariates at baseline, the hazard ratios of pneumonia death were 1.30 (95%CI:0.64-2.70) for pre-diabetics and 2.28 (95%CI:1.18-4.39) for diabetics, respectively. Among those covariates, only age (HR (95%CI);1.16 (1.13-1.20)), gender as female (0.35 (0.22-0.61)) and physical fitness measured as having no problem walking 1+mile during the previous month (0.38 (0.20-0.67)) reached statistical significance. Conclusions The results suggest that diabetes mellitus is a strong risk predictor of pneumonia mortality and the evaluation of physical fitness may also be useful in the risk prediction of pneumonia mortality for seniors.

BACKGROUND:There is a certain relationship between bone marrow mesenchymal stem cels and articular cartilage, and its celular functional status is one of the causes of osteoarthritis.
OBJECTIVE:To explore the correlation between functional status of bone marrow mesenchymal stem cels and disease progression in patients with osteoarthritis.
METHODS:The clinical data of 45 patients were colected, including 15 cases undergoing arthroscopic knee cruciate ligament reconstruction (control group), 15 cases of mild osteoarthritis (mild osteoarthritis group), 15 cases of severe osteoarthritis (severe osteoarthritis group). Bone marrow mesenchymal stem cels from three groups were culturedin vitro to observe the morphological changes. Proliferative and differentiation ability was detected and compared between the three groups.
RESULTS AND CONCLUSION:At 24 hours of primary culture, the bone marrow mesenchymal stem cels were arranged in regular pattern and spindle-shaped in the three groups with no morphological differences. At 14 days of chondrogenic induction, toluidine blue staining was positive for bone marrow mesenchymal stem cels, but there was stil no difference in the chondrogenic ability in the three groups. Cel counting kit-8 results showed that the proliferative ability of cels was strongest in the control group and weakest in the severe osteoarthritis group. These findings indicate that there is a certain correlation between the functional status of bone marrow mesenchymal stem cels and the progress of the disease.

S100A7 protein has a variety of functions,concerning with multiple basic biological processes of cells such as proliferation,differentiation,apoptosis,signal transduction,lipid metabolism,adherence and migration.It is expressed mainly in the inflammatory diseases and tumors,and strongly correlated with metastasis of some tumors.

Objective:To study the relationship between serum uric acid level and hyperlipidemia.Methods:The serum uric acid level between hyperlipidemia group (103 cases) and control group (109 cases) was compared.The correlation of serum uric acid level and hyperlipidemia was analysed using multiple stepwise regression model.The logistic regression analysis was used to investigate the association of serum uric acid level with the incidence of hyperlipidemia.Results:The serum uric acid level in hyperlipidemia group[(382.4?100.3)?mol/L] was significantly higher than in control group[(351.7?88.9)?mol/L]( P 0.05).Conclusion:The serum uric acid level increases in the hyperlipidemia patients, and is closely related to blood lipid, but it is not an independent risk factor for hyperlipidemia.

Objective:To study the relationship between serum uric acid level and coronary heart disease.Methods:212 patients in hospital were divided into two groups of non-coronary heart disease (91 cases) and coronary heart disease (121 cases).The group of coronary heart disease was divided into three subgroups of single-branch,double-branch and three-branch according to the scope of pathological changes.The serum uric acid levels in different groups were compared.The correlation between serum uric acid level and coronary artery lesion was analysed using univariate stepwise regression model.The logistic regression analysis was used to investigate the association of serum uric acid level with the incidence of coronary heart disease.Results:The serum uric acid level in coronary heart disease group[(378?100)?mol/L]was significantly higher than that in non-coronary heart disease group[(345?77)?mol/L](P0.05).Conclusion:The serum uric acid level increases along with the severity of coronary heart disease, but it is not an independent risk factor for coronary heart disease.

Objective:To evaluate the efficacy of coronary stenting in the treatment of patients with unstable angina.Methods:38 patients with unstable angina underwent coronary stenting based on routine conservative therapy.The clinical outcome was evaluated with regard to the frequency of angina and the changes in electrocardiogram.The patients were followed up for average 6 months.Results:The procedural success rate was 94.7% in the patients with residual stenosis less than 10% and initial TIMI grade 3 flow.There were no recurrent angina in 36 patients and significantly less angina frequency in 2 patients.The electrocardiogram in 29 patients recovered normally.More than 50% ST-T segment changes happened in 5 patients while 4 patients suffered no obvious changes.During the follow-up,4 patients presented recurrent angina,3 of which underwent repeat coronary stenting.There was no acute myocardial infarction or death.Conclusion:Coronary stenting is a safe and effective treatment for unstable angina.

Objective To compare short- and long-term outcome of rehabilitation in different age patients with severe traumatic brain injury. Methods 116 patients with severe traumatic brain injury aged 18~65 years were divided into 18~44 years old group and 45~65 years old group. They were assessed with Mini-Mental Status Examination (MMSE), simplified Fugl-Meyer Motor Functional Assessment, Fugl-Meyer Balance Assessment, modified Barthel Index (MBI) and Disability Rating Score (DRS) before and 3 months after rehabilitation. And they were followed up with DRS 1 year after discharge. Results All the indexes improved 3 months after rehabilitation (P<0.01). After 1 year of follow-up, the DRS scores continued to decline in both groups, especially in the 18~44 years old group (P<0.05). Conclusion Early rehabilitation can improve the limb function and activities of daily living of adult patients with severe traumatic brain injury, especially in younger people.

Objective To evaluate the efficacy and safety of Xiao-fan-xi-ji in treatment of the post operation of mixed hemorrhoids. Methods198 patients were randomly divided into the treatment group(112 cases,treated by Xiao-fan-xi-ji for 15 days)and the control group(96 cases,treated by Potassium Permanganate for 15 days).The improvement of symptom,sign and quality of life scores were evaluated. ResultsThe experimental and control groups in age,sex 、ethnic and the degree of disease had no statistical significance(all P＞0.05).After 2,7 days treatment,major adverse reaction was observed.Compared two groups in postoperative pain and edema edge,there was significant difference between the two groups(P＜0.05).The average healing time in experimental group was shorter than control group.Compared with control group,the quality of life scores increased after the treatment,the differences had statistical significance(P＜0.05). ConclusionXiao-fan-xi-ji was effective in the treatment of patients with mixed hemorrhoids after the operation,it was worth being promoted.

Drug-drug interaction (DDI) is referred as the changes of physical and chemical properties, as well as the pharmacokinetics or pharmacodynamics of drugs administered simultaneously or consecutively. The clinical results for drug-drug interaction could be divided into good clinical efficacy and adverse interaction. With the kinds of drugs increasing every year, new drug resistances spring up frequently. This phenomenon makes drug combination increased so that the drug interaction, especially the adverse interaction emerged. The mechanisms of in vivo drug-drug interaction are relevant to a number of factors, including drug-metabolizing enzyme systems and membrane transporters. Recent studies have revealed the important role played by transporters in drug absorption, distribution, metabolism and elimination. In order to avoid severe side effects mediated by transporters and to promote rational combination in clinics, the mechanisms of intestinal absorption and renal excretion mediated by transporters are reviewed.