The cis-epoxysuccinate hydrolase (CESH) from Rhizobium strain BK-20 is the key enzyme for L(+)-tartaric acid production. To establish a highly efficient and stable production process, we first optimized the enzyme production from Rhizobium strain BK-20, and then developed an immobilized cell-culture process for sustained production of L(+)-tartaric acid. The enzyme activity of free cells reached (3 498.0 +/- 142.6) U/g, and increased by 643% after optimization. The enzyme activity of immobilized cells reached (2 817.2 +/- 226.7) U/g, under the optimal condition with sodium alginate as carrier, cell concentration at 10% (W/V) and gel concentration at 1.5% (W/V). The immobilized cells preserved high enzyme activity and normal structure after 10 repeated batches. The conversion rate of the substrate was more than 98%, indicating its excellent production stability.
Alginates ; chemistry ; Cells, Immobilized ; Glucuronic Acid ; chemistry ; Hexuronic Acids ; chemistry ; Hydrolases ; metabolism ; Rhizobium ; enzymology ; metabolism ; Tartrates ; metabolism

Objective To explore the clinical features, diagnosis, and treatment of neonatal necrotizing pneumonia. Methods The clinical data of two cases of neonatal necrotizing pneumonia were retrospectively analyzed. The clinical features, diagnosis, and treatment of neonatal necrotizing pneumonia in literatures were summarized. Results Two cases were diagnosed of community-acquired Staphylococcus aureus necrotizing pneumonia and had the onset with fever. The chest X-ray showed exudative change with cystic shadow. The chest CT showed multiple cavity changes. The sputum and blood cultures were positive for Staphylococcus aureus. Both of them were effectively treated by vancomycin. The imaging was improved during the follow-up. Searching the database, 4 related literatures were being found, and there were totally 7 cases of neonatal necrotizing pneumonia including current 2 cases. The main features were as follows: The pathogenic bacteria in all cases include Staphylococcus aureus. One case was combined with pseudomonas aeruginosa. Six cases were community-acquired infections. All of them were non-immune deficiency newborn. Six cases were primary necrotizing pneumonia. Six cases were unilateral lung involvement. Five cases got fever, 5 cases had septicemia, 3 cases had pleural effusion, 2 cases had aerothorax, one case had bronchial chest and 2 cases had extrapulmonary infection. The C-reactive protein was increased in all cases. Three cases need mechanical ventilation. Six cases had a good prognosis. Conclusions The main pathogenic bacterium in neonatal necrotizing pneumonia was Staphylococcus aureus. The diagnosis was mainly depends on the typical imaging and pathogenic examination. The treatment is mainly the use of antibiotic for gram positive cocci.

BACKGROUND: It has been proved that platelet activation is involved in the development of diabetic angiopathy. Glycoprotein (GP) Ib/IX/V complex is one of the main platelet membrane glycoproteins, and the receptor of both von Willebrand Factor and thrombin. It plays a key role in the process of platelet activation.OBJECTIVE: To observe the expression changes of GP Ib/IX/V complex and its component GP Ibα in patients with type 2 diabetes mellitus. DESIGN: Case-control study. SETTING: Department of Integrated Chinese and Western Medicine, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology.PARTICIPANTS: A total of 51 type 2 diabetic outpatients who visited Union Hospital were enrolled from December 2005 to January 2007. The diagnosis was based on the independent criteria from WHO in 1999. Of all the 51 patients, there were 23 females and 28 males, with a peripheral platelet count of over 50×109 L-1. All the subjects had no history of administrating drugs two weeks before the examination, which would potentially influence platelet count. According to disease controlling condition, the patients were assigned to well controlled patient (WCP) group (n = 25) and poorly controlled patient (PCP) group (n =26); and according to whether angiopathy was accompanied, diabetic patients were divided into vascular disease (VD) group (n =27) and non-vascular disease (NVD) group (n =24). Meanwhile 23 healthy subjects were enrolled as normal control group. Informed consents were obtained from subjects and their relatives. The experiments were approved by the ethical committee of Union Hospital.METHODS: Fasting venous blood was harvested from all the subjects' elbow on the early morning of visit day.①Biochemical analysis: Fasting plasma glucose (FPG), glycosylated hemoglobin (HbA1c) and fasting plasma insulin (FINS) was measured by the Clinical Chemistry Department in Union Hospital.② Measurement of platelet membrane GP Ib/IX/V complex and its component GP Ibα: First, 3 mL cubital fasting blood was drawn from each subject and was anti-coagulated with 38 g/L natrium citricum. After that, all samples were fixed with 10 g/L paraform for 45 minutes. Then 50 μL well-fixed blood was added into the polystyrene tube, meanwhile 20 μL monoclonal antibody, such as CD42a-b-c-d and PE-labeled CD42b, was respectively mixed gently with the blood sample and incubated at room temperature in dark for 30 minutes. Next, 20 μL FITC-labeled rat IgG was mixed with the sample containing CD42a-b-c-d and incubated equally. In the end all blood samples were analyzed by FACS420 flow cytometry and the results were expressed as mean fluorescence intensity (MFI). ③ Platelet maximun aggregation rate (MAR) was detected according to reference.MAIN OUTCOME MEASURES: ①Biochemical indicators;②The expressions of GP Ib/IX/V complex and GP Ibα;③Platelet MAR.RESULTS: Fifty-one patients with type 2 diabetes mellitus and twenty-three healthy subjects were all involved in the result analysis.①There were significant differences in FINS in WCP group and PCP group compared with normal controls (P ＜ 0.01). FPG and HbA1c were significantly higher in PCP group compared with normal control group and WCP group (P ＜ 0.01).②Expressions of GP Ib/IX/V complex and GP Ibα were significantly lower in WCP group and PCP group compared with normal control group (P ＜ 0.01), significantly lower in PCP group than in WCP group (P ＜ 0.05), and also significantly lower in VD group and NVD group compared with normal control group (P ＜ 0.01). Moreover the expression of GP Ibα in VD group and NVD group was significantly lower than that of normal control group (P ＜ 0.05), and it also significantly decreased in VD group compared with NVD group (P ＜ 0.05). MFI of GP Ib/IX/V complex had an obvious negative correlation with FBG, HbA1c and FINS (r =-0.634, -0.573, -0.649, P ＜ 0.05), and GP Ibα MFI was obviously negatively correlated with FBG and HbA1c (r =-0.602, -0.543, P ＜ 0.05).③Platelet MAR of diabetic patients were remarkably higher than in healthy subjects (t =-3.852, P ＜ 0.01). Platelet MAR in PCP and VD groups were respectively higher than those in WCP and NVD groups (P ＜ 0.05). CONCLUSION: Platelet activation exists in the early stage of type 2 diabetes mellitus without diabetic angiopathy, and is more obvious after diabetic angiopathy. There is a positive correlation between platelet activation and blood glucose. As a receptor of thrombin and von Willebrand Factor, GP Ib/IX/V complex may be involved in the development of diabetic angiopathy.

BACKGROUND: The peripheral blood stem cell is a multi-differentiation precursor cell, and it can differentiate into osteoblasts. Tissue engineered bone, which is regarded as a vector of cell transplantation, has good compatibility with receptor tissue and seed cells. Transforming growth factor-β (TGF-β) is an important regulatory factor for repairing bone injury. Additionally, TGF-β can induce peripheral blood stem cells to differentiate and proliferate into osteoblasts.OBJECTIVE: To study TGF-β expression in repairing alveolar bone during synergetic transplantation of peripheral blood stem cells and tissue engineered bone. DESIGN: Observational study.SETTING: Stomatology Hospital of Xi'an Jiaotong University.MATERIALS: This study was performed at the Stomatology Hospital of Xi'an Jiaotong University from 2003 to 2006. Experimental animals were provided by the Animal Experimental Center, Medical College of Xi'an Jiaotong University (original Xi'an Medical University). All animals were intramuscularly induced with ketamine, intramuscularly anesthetized with sumianxin, and then sacrificed for surgery. The experiment was approved by the local ethics committee.METHODS: Peripheral blood stem cells were extracted from dog and prepared as a cell suspension. Iliac bone was obtained from healthy pig to prepare decalcifying-deproteinic tissue engineered bone. The tissue engineered bone was then dipped into peripheral blood stem cell suspension. Ten healthy hybrid dogs were randomly divided into an experimental group and a control group, with 5 dogs in each group. An incision was made from left to right along the canine teeth of the lower mandible, along the lip, lateral to the gingival sulcus, to the alveolar crest, and then along the bilateral vestibular groove to form a trapezoid segment. Subsequently, the segment was turned downward to expose the bone lamella lateral to the lip. In addition, a bone defect region of 2 cm × 2 cm × 1 cm was drilled between the lateral incisor of lower mandible using a turbine drill. Peripheral blood stem cell-tissue engineered bone was implanted in the experimental group but tissue engineered bone only was implanted in the control group. At 2, 3, 4, 8 and 12 weeks after surgery, during the differentiation and proliferation of peripheral blood stem cell into osteoblasts, TGF-β expression was measured using immunohistochemistry.MAIN OUTCOME MEASURES: ① Morphological changes of peripheral blood stem cells differentiating into osteoblasts and structural function of organoid were observed under optical microscopy and by transmission electron microscopy. ② TGF-β expression was measured using immunohistochemistry during the differentiation and proliferation of peripheral blood stem cells into osteoblasts.RESULTS: Two weeks after peripheral blood stem cell-tissue engineered bone transplantation in the experimental group, TGF-β expression was mildly positive at the fringe of the bone defect. Four to eight weeks after the transplantation, high numbers of osteoblasts, fibroblasts and collagenous fibers were found at the center of the bone defect region, and TGF-β expression was strongly positive. The bone defect was completely repaired after 12 weeks. In the control group, 8-12 weeks after tissue engineered bone transplantation, TGF-β expression was mildly positive only at the fringe of the bone defect. CONCLUSION: During dog alveolar bone defect repair, TGF-β can induce peripheral blood stem cells, in combination with tissue engineered bone, to differentiate and to proliferate into osteoblasts.

Objective To express and characterize the virus-like particles( VLPs) of H5 subtype containing of hemagglutinin ( HA ) and matrix 1 ( M1 ) protein by using Baculovirus-insect cells .Methods Full length genes encoding HA protein from the A/Indonesia/05/2005(H5N1) strain and the M1 protein from the A/Anhui/01/2005 ( H5N1 ) strain were cloned into a baculovirus expression vector to construct pFBD-M1-HA.The expression of HA and M1 proteins were detected by Western blot and indirect immunoflu-orescence after the transfection of Spodoptra frugiperda (Sf9) insect cells with recombinant baculovirus.Pu-rified VLPs were analyzed by SDS-PAGE and visualized with transmission electron microscope.The biologi-cal activity of purified VLPs was detected by hemagglutination test.Results The HA and M1 proteins of H5 subtype expressed by baculovirus-insect cells could be self-assembled into the functional mature VLPs.The hemagglutination titer of VLPs was as high as 1024 HAU/50μl.Conclusion The H5 subtype VLPs as pre-pared in this study would pave a way for the development of a candidate recombinant A ( H5) vaccine.

OBJECTIVETo analysis chemical components of volatile oil from the seed of Nigella glandulifera (NG), comparing them with those from the seed of foreign N. sativa (NS) and N. damascene (ND), and to quantify thymoquinone in the volatile oil extracted by hydrodistillation (HD) from the seed of NG.

METHODThe volatile oil was extracted by supercritical CO2 extraction (SFE-CO2 ) and HD from the seed of NG and its chemical components was analysed by GC-MS, the relative percentage of components were determined by peak aera normalization method and compare with those of the seed of NS and ND. The content of the thymoquinone in the volatile oil was determined by one point external standard method.

RESULTIn terms of the volatile compounds, p-cymene is the major component of NG and NS, their relative percentage contents are 33.75% and 61.48%, respectively. beta-Elemene is the major component of ND, its relative percentage content is 73.24%. The relative percentage contents of thymoquinone are 3.73% (HD), 3.80% and 0.08% in NG, NS and ND, respectively. Linoleic acid is a major component of volatile oil by SFE-CO2 in NG, but its content of p-cymene is lower. The absolute percentage content of thymoquinone is 1.58% by HD in volatile oil of NG.

CONCLUSIONThere are comparatively large differences of volatile components in NG, NS and ND.

Benzoquinones ; analysis ; Nigella ; chemistry ; Oils, Volatile ; analysis ; Plant Oils ; analysis ; Seeds ; chemistry

Objective To investigate correlation between aspirin resistance(AR) and inflammatory factors. Methods One hundred and ten patients with coronary heart disease took aspirin 0.1 mg/d for 14 days.It was detected platelet aggregation function induced with adenosine disphosphate (ADP) and arachidonic acid (AA), and investigated correlation between AR and inflammatory factors. Interleukin-1? (IL-1?),interleukin-6 (IL-6) and high sensitive C-reaction protein (hs-CRP) levels. Results IL-6 level of patients with AR was significantly higher than that of aspirin sensitive (AS) patients. The other two index were not different between the two groups. Conclusion IL-6 levels could be used as predictor.

Objective To explore the temporal trend of cancer death rates in different age and the influencing factors in Kunshan,Jiangsu province,1981 to 2015.Methods Data were derived from cancer rcgistry and vital registration system.The Chinese age structure in 2000 was used to calculate age-standardized death rates (ASR),and annual percentage changes (APC) and 95％ confidence interval (Cl) were used to estimate the temporal trend of cancer death rates.Difference decomposition method was applied to analyze the contribution of demographic and non-demographic factors for the change of cancer mortality.Results Between 1981 and 2015,the age standardized all cancers death rate decreased from 162.49 to 93.74 per 100,000 (APC=-l.6％,95％ CI:-1.8％--1.5％).However,the ASR for those aged 70 years or above was stable over time (APC=0.2％,95％ CI:-0.2％-0.5％),whereas aged 30-69 years was decreased from 240.01 in 1981 to 93.28 in 2015 (APC=-2.8％,95％ CI:-3.0％--2.6％).In addition,the proportion of leading cancers were changed obviously.The proportion of lung cancer increased from 1981 to 2015,while gastric cancer,liver cancer,esophageal cancer and colorectal cancer decreased.Compared with the crude cancer mortality in 1993,the effect of the demographic and non-demographic factors to the increased death rate in 2015 were 308.93％ and-208.93％,respectively.Conclusion The ASR death rate of all cancers was decreasing,and the rate in those aged 30 to 69 years decreased significantly,whereas stable in those aged 70 years or above.The effect of demographic characteristics on cancer mortality was significantly greater than that of non-demographic characteristics.

OBJECTIVETo estimate the dose response relationship between cigarette smoking and hypertension in men based on restricted cubic spline method.

METHODSUnder the proportion to the population size, 38 520 subjects were randomly selected from May to August 2012 with cluster sampling method in urban and rural areas of Kunshan, China.Each participant received face-to-face interview with the standardized questionnaire, and physical examination.Restricted cubic spline was employed to estimate the dose response relation of cigarette smoking on the risk of hypertension.

RESULTSThe overall prevalence of cigarette smoking was 22.6% (8 691/38 520), prevalence of cigarette smoking in men was 46.1% (8 499/18 454). Multiple logistic regression analysis showed that current smoking (OR = 1.16, 95%CI:1.05-1.28) and previous cigarette smoking (OR = 1.32, 95%CI:1.07-1.63) were associated with hypertension after adjusted confounding factors (age, sex, body mass index, education, family income per month, urban or rural areas, physical activity and physical exercise) in men. After further adjusting drinking status, only previous cigarette smoking was associated with hypertension (OR = 1.28, 95%CI:1.04-1.58). The restricted cubic spline model indicated a linear dose-response relation between hypertension and cigarette smoking per day in men (non-linearity test P = 0.604 1).However, a non-linear dose response relation was found between duration of smoking (non-linearity test P < 0.000 1), smoking index (non-linearity test P = 0.009 9) and hypertension.

CONCLUSIONLong-term and heavy cigarette smoking is associated with hypertension in men.

Adult ; Aged ; Alcohol Drinking ; Body Mass Index ; China ; epidemiology ; Humans ; Hypertension ; epidemiology ; Male ; Middle Aged ; Prevalence ; Risk Factors ; Smoking ; epidemiology ; Surveys and Questionnaires