The strain of Cunninghamella echinulata 9980 was first selected with high aminoacylase activity . In three submerged cultures, the aminoacylase activity in the mycelial cell was compared . A number of factors have effects on the resolution reaction. The results showed that, peptone culture gave the highest aminoacylase activity with 680U/g.The optium temperature,pH,and substrate concentration were 55℃, 7.0,and 0.2mol/L,respectively. The ions in the buffer lowered the activity,but the Co~(2+) in 10~(-4)~10~(-3 )mol/L was necessary for its activity.

Arginine Deiminase(ADI) was purified to homogeneity using ammonium sulfate precipitation,Q-Sepharose Fast Flow anion exchange chromatography and SephadexG-75 gel filtration chromatography. This purification protocol resulted in a 34.5-fold purification of ADI with 31.4% final yield. A molecular weight of about 190 kD determined by native gradient polyacrylamide gel electrophoresis. The enzyme has only one kind of 46 kD subunit determined by SDS-PAGE. Combining the results from the two kinds of electrophoresis,the authors deduce that the enzyme may be a tetramer. The optimum pH and temperature for lipolytic activity of ADI was pH 6.5 and 50℃,respectively. It was extremely stable at 45℃ and retained 97.9% of its original activity for 30 min. The stability declined rapidly as soon as the temperature rose over 50℃. ADI was highly stable in the pH range from pH 5-8. ADI acted on L-arginine but not on D-arginine. ADI catabolism was dependent on metal ions. At their adequate concentration,Mn2+,Mg2+ and Co2+ were the effective promoter,while superfluous Zn2+and Co2+ inhibited ADI activity. L-citrulline did not act on ADI,but L-ornithine inhibited ADI activity. The degradation of L-arginine with ADI catalysis was according to simple Michaelis-Menten equation. The Michaelis constant was 3.2686 mmol/L and the maxi-mum velocity was 2.44 ?mol/min.

L-theanine (γ-glutamylethylamide) is the main free amino acid component of tea and its favorable physiological effects on mammals have been reported. An enzymatic method for optically pure L-theanine production with a new L-aminoacylases-production fungi Cunnighamella echinulata 9980 was developed. The optimum conditions for enzymatic catalysis were at pH 6.5 with 50 mmol/L N-Acyl-DLtheanine and 40 mmol/L CoCl2. After 12-h incubation at 50℃,22.5 mmol/L L-theanine was obtained, the conversion rate against N-Acyl-L-theanine being 90%. Cunnighamella echinulata and the aminoacylase were applied in preparation of L-theanine.

OBJECTIVEThis preliminary study aims to investigate the effects of titanium and titanium alloy micro-nano-dimensional topography on the biological behavior of osteoblasts in vitro.

METHODSElectrolytic etching (EE) method was used to produce micro-nano dimensional titanium surfaces. The surfaces were observed to determine their effects on the adhesion, proliferation, cell morphology, and alkaline phosphatase (ALP) activity of osteoblasts.

RESULTSThe surfaces of the titanium and titanium alloy groups exhibited higher adhesion and proliferation of osteoblasts than those of the mechanical group. The titanium surface was covered with a group of cells, a large number of filopodia, and functional particles. The ALP activity of the titanium group was significantly higher than that of the titanium alloy and mechanical groups.

CONCLUSIONEE method in pure titanium and titanium alloy surfaces result in bowl-like nests and nanostructures of different diameters and depths. The diameters of the pure titanium and titanium alloy surfaces range from 30 to 50 μm and 5 to 8 μm, respectively. The former is more conducive to promote the proliferation and differentiation of cells.

Alloys ; Cell Differentiation ; Cell Proliferation ; Dental Etching ; Humans ; Nanostructures ; Osteoblasts ; Prostheses and Implants ; Surface Properties ; Titanium

OBJECTIVESome neonates especially premature infants, low birth weight infants and extremely low birth weight infants have limited endogenous energy stores. It is necessary to establish continuous administration of postnatal nutrition. The use of parenteral nutrition (PN) in neonates with immaturity of digestive system and intentionally delayed feedings has gained widespread acceptance. PN has been shown to provide sufficient nutrients to maintain growth in newborn infants. The major complication of PN in neonates is PN-associated cholestasis (PNAC). It remains a significant and frequent clinical problem for neonatal practitioners. In some cases, progressive liver damage, liver failure and death may become inevitable. In order to analyze the risk factors of the PNAC in neonates and to provide the evidence of safety and efficiency in clinical nutrition support, the clinical data of 612 neonates who had received PN for more than 5 days during the past 20 years were reviewed.

METHODSRetrospective analysis on data collected from April 1985 to March 2005 was performed. The records of 612 neonates were divided into two groups according to the established Nutrition Support Team (NST) in our hospital. Each group included two sub-groups. Seventy neonates of the first group were divided into PNAC group (n = 6) and non-PNAC group (n = 64); these patients were seen between 1st April 1985 and 31st March 1995. The remaining 542 neonates of the second group who were also divided into 2 groups, i.e. PNAC group (n = 12) and non-PNAC group (n = 530) who were seen from 1st April 1995 through 31st March 2005. The incidence of PNAC between the first group and the second group was compared and the associated factors were analyzed. The PNAC was defined when serum level of direct-bilirubin exceeded 1.5 mg/dl or direct-bilirubin greater than 50% of the bilirubin and excluding cholestasis resulted from other diseases.

RESULTSThe total incidence of PNAC in neonates who had received TPN for more than 5 days was 2.94%. The incidence of PNAC of the first and the second decade was 8.57% and 2.21%, respectively (OR = 0.242, 95% CI = 0.088 approximately 0.666). The average gestational age (GA) and birth weight (BW) of PNAC group were less than those of the non-PNAC group (GA: (33 +/- 5) w vs. (36 +/- 4) w, P = 0.009; OR = 0.827, 95% CI = 0.698 approximately 0.980. BW: (2003 +/- 743) g vs. (2393 +/- 764) g, P = 0.045; OR = 1.001, 95% CI = 0.999 approximately 1.002). The PN duration and calorie intake of PNAC group was longer than that of the non-PNAC group (PN duration: 32 +/- 30 d vs. (13 +/- 10) d, P = 0.000; OR = 1.072, 95% CI = 1.032 approximately 1.112. Calorie intake: [(272 +/- 46) kJ/(kg.d)] [(65.0 +/- 10.9) kcal/(kg.d)] (1 kcal = 4.184 kJ) vs. [(232 +/- 55) kJ/(kg.d) (55.5 +/- 13.1) kcal/(kg.d)], (P = 0.002; OR = 1.066, 95% CI = 1.012 approximately 1.122), but the weight gain in the non-PNAC group had a tendency to increase as compared to that of the PNAC group [(20 +/- 27) g/d vs. (9 +/- 19) g/d, P = 0.175].

CONCLUSIONSThe incidence of PNAC was associated with the longer duration of PN, the smaller age at initiation of PN, the higher calorie intake, prematurity and lower birth weight. Establishment of the nutrition support team can normalize the practice of the PN administration and decrease the incidence of the complication with nutrition support. It is a favorable mode and it can provide a safer, more effective and reasonable means in clinical nutrition support. To avoid PNAC, it is suggested that the administration of enteric feeding should start as soon as possible, which may enhance effective contraction of gallbladder and secretion of gastrointestinal hormones, and it is best to avoid high calorie of PN and control the calorie intake under 251.04 approximately 334.72 kJ/(kg.d) [60 approximately 80 kcal/(kg.d)].

Cholestasis ; complications ; epidemiology ; etiology ; Female ; Gestational Age ; Humans ; Incidence ; Infant ; Infant, Low Birth Weight ; physiology ; Infant, Newborn ; Infant, Premature ; growth & development ; Male ; Parenteral Nutrition ; adverse effects

Objective To analyze the stage characteristics in the exercise behavior improvement of college students and explore the role of Process in the exercise behavior change based on the transtheoretical model, providing basis for the stage-matched intervention for the exercise behavior of college students. Methods There were 932 students who completed the questionnaires, from 5 universities in Shanxi Province were selected by using a stratified random cluster sampling method. Descriptive analysis was used to describe the exercise behavior of college students. Variance analysis and multivariate Logistic regression model were used to analyze the difference of the process of with stage of change among college students’ physical exercise. Multivariate variance analysis was used to analyze how personal characteristics affect process of change. Results Among all participants, 89.4% students knew the importance of physical exercise, and 29.4% students were satisfied with their physical exercise condition. The distribution of students’ physical exercise stage showed an inverted U-shape with left-side peak, and there was a significant difference between gender( 2=54.657, P<0.001). There were significant stage characteristics in the process of students' exercise behavior, gender had a significant main effects on mutual aid relation(F=7.400, P=0.07)and conscious control (F=7.778, P=0.005), gender and grade had interaction effects on social release (F=3.614, P=0.013). Conclusions The college students' exercise behavior showed the characteristics of “knowing but not to do”, which conformed to the Transtheoretical model. It is essential to develop targeted phased exercise intervention strategies according to the relationship between change of phase and change of procedure.

[Objective ] To ascertain the current situation in application of summary procedure within the jurisdiction , and propose corresponding measures and suggestions for prevention of administrative reconsideration and administrative lawsuits resulting from improper use of summery procedure . [ Meth-ods] Retrospective examination was done of 63 administrative penalty cases by summary procedure in Xu-hui District in 2011 and 2012 .Some parties punished were visited , and at the same period were randomly selected the penalty cases from the same professional use and general procedure .Control analysis was done from six aspects such as punishment body identified , confirmation of illegal facts , fixation of evidence , ap-plication of legal provisions , legitimate punishment procedure , clerical production specification and others . [ Results] Problems were found to be existent in the six aspects of punishment body identified , confir-mation of illegal facts , fixation of evidence , application of law provisions , legitimate punishment proce-dures, and clerical production specification , etc.There is a certain gap with the general procedure punish-ment cases , which needs to be improved in future . [ Conclusion ] Administration by law awareness should be strengthened , training in this regard enhanced , management norms for summary procedure formu-lated , evidence system for summary procedure established and perfected , and inspection strengthened , so as to improve the quality of summary punishment cases .

Background/Aims: Metabolic risk factors could accelerate hepatitis B virus (HBV)-related mortality; however, their impacts on disease severity in HBV-related acute on chronic liver failure (HBV-ACLF) patients remain unexplored. In this study, we assessed the effects of metabolic risk factors on the outcome of HBV-ACLF patients. Methods: This study retrospectively enrolled antiviral therapy naïve HBV-ACLF patients from a single center in China. Patients were evaluated according to Child-Turcotte-Pugh score, Model for End-Stage Liver Disease (MELD) score, 30-day, 90-day mortality and survival rate to estimate the prognosis of HBV-ACLF. The impacts of different metabolic risk factors were further analyzed. Results: A total of 233 patients, including 158 (67.8%) with metabolic risk factors and 75 (32.2%) without metabolic risk factors, were finally analyzed. Patients with metabolic risk factors had significantly higher MELD score (22.6±6.1 vs 19.8±3.8, p<0.001), 90-day mortality rate (56.3% vs 38.7%, p=0.017), and shorter median survival time (58 days vs 75 days: hazard ratio, 1.553; 95% confidence interval, 1.061 to 2.274; p=0.036) than patients without them. Moreover, metabolic risk factors were independently associated with patients’ 90-day mortality (hazard ratio, 1.621; 95% confidence interval, 1.016 to 2.585; p=0.043). Prediabetes/diabetes and hypertension were related to higher rates of infection and worse renal function in HBV-ACLF patients. Conclusions HBV-ACLF patients with metabolic risk factors, especially prediabetes/diabetes or hypertension, could have more severe disease and lower survival rates. In addition, the existence of metabolic disorder is an independent risk factor for HBV-ACLF patients’ 90-day mortality.

OBJECTIVETo investigate the effect of farnesoid-X-receptor (FXR) antagonist Z-guggulsterone in an in vivo high-fat fed apolipoprotein E knockout (ApoE(-/-)) mice model of myocardial ischemia/reperfusion (I/R).

METHODSMale ApoE(-/-) mice were randomly divided into three groups: standard ApoE(-/-) group (fed with standard mouse diet for 12 weeks before myocardial I/R procedure, n = 18), high-fat ApoE(-/-) group (fed with high-fat mouse diet for 12 weeks before myocardial I/R procedure, n = 22), and high-fat ApoE(-/-) + FXR antagonist group(fed with high-fat mouse diet for 12 weeks and received FXR antagonist Z-Guggulsterone 30 minutes before myocardial I/R procedure, n = 17). The expression of FXR was detected by real-time quantitative-PCR. Myocardial infarct size was determined by Evans blue/TTC double staining methods. Myocardial apoptosis was determined by in situ TUNEL technique. Markers of the mitochondrial-mediated apoptotic pathway (cytochrome c release, caspase-9 activity, and BAX and BCL-2 levels), endoplasmic reticulum stress apoptotic pathway (caspase-12 activity and CHOP level), and death receptor apoptotic pathway (caspase-8 activity, and Fas and FasL levels) were also measured.

RESULTFXR expression (3.7-fold higher, P < 0.01), myocardial infarct size [(62.1 ± 7.0)% vs. (33.8 ± 5.8)%, P < 0.01] and myocardial apoptosis index[ (36.8 ± 5.7)% vs. (17.2 ± 3.8)%, P < 0.01]were all significantly higher in high-fat ApoE(-/-) group than those in standard ApoE(-/-) group. Compared with high-fat ApoE(-/-) group, myocardial infarct size [(24.4 ± 4.7)% vs. (62.1 ± 7.0)%, P < 0.01] and myocardial apoptosis index [(13.8 ± 2.7)% vs. (36.8 ± 5.7)%, P < 0.01] were significantly reduced in high-fat ApoE(-/-) + FXR antagonist group. Moreover, levels of mitochondrial-mediated apoptotic pathway markers (cytochrome c release, caspase-9 activity, and BAX/BCL-2 levels) and endoplasmic reticulum stress apoptotic pathway markers (caspase-12 activity and CHOP level) were significantly lower in high-fat ApoE(-/-) + FXR antagonist group than those in high-fat ApoE(-/-) group (all P < 0.01). Levels of death receptor apoptotic pathway markers (caspase-8 activity, and Fas and FasL levels) were similar between high-fat ApoE(-/-) group and high-fat ApoE(-/-) + FXR antagonist group.

CONCLUSIONFXR antagonist alleviates myocardial reperfusion injury in cholesterol-fed ApoE(-/-) mice via inhibition of the mitochondrial-mediated and endoplasmic-reticulum stress pathway.

Animals ; Apolipoproteins E ; genetics ; Apoptosis ; drug effects ; Caspase 9 ; metabolism ; Cholesterol, Dietary ; administration & dosage ; Cytochromes c ; metabolism ; Disease Models, Animal ; Endoplasmic Reticulum Stress ; Male ; Mice ; Mice, Knockout ; Myocardial Reperfusion Injury ; metabolism ; pathology ; prevention & control ; Pregnenediones ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Receptors, Cytoplasmic and Nuclear ; antagonists & inhibitors ; metabolism ; bcl-2-Associated X Protein ; metabolism

OBJECTIVETo explore the relationship between expression thange of P-selectin after brain injury and secondary brain damage.

METHODSSixty SD rats were randomized into 3 equal groups, namely the control group, mild injury group and severe injury group and animal models of brain injury were established in SD rats according to the method of Feeney. P-selectin expression in the brain tissues were determined at 6 h and l, 3, and 7 days following brain injury (n=5 for each time point). Imaging analysis was performed using computerized imaging technique.

RESULTSP-selectin expression and neutrophil infiltration in the brain tissues increased significantly 6 h after brain injury (P<0.05), reaching the peak level at postoperative 24 h and then gradually decreased.

CONCLUSIONP-selectin expression and neutrophil infiltration increase significantly following brain injury, and the time course and distribution of P-selectin expression are consistent with the secondary damage of the brain, strongly suggesting the involvement of P-selectin upregulation in the secondary insult after brain injury.

Animals ; Brain Chemistry ; Brain Injuries ; etiology ; metabolism ; pathology ; Female ; Immunohistochemistry ; P-Selectin ; biosynthesis ; Random Allocation ; Rats ; Rats, Sprague-Dawley