The culture conditions of Saccharomyces cerevisiae sp.strain by 1.1 b were optimized for the production of D-(-)-mandelate dehydrogenase which is useful for the asymmetric bioreduction of benzoylformate to form D-(-)-mandelate.The optimum medium(per liter)consistes of 60 g peptone,30 g maltose, 0.5 g MgSO_4,0.01 g ZnSO_4,1.0 g KCl.After optimization of the culture medium,the enzyme production in shake flasks is enhanced from 2.56 to 20.21 U/L.The optimum fermentation conditions were determined as follows:medium volume 100 mL(i.e.,40%for a 250-mL shake flask),pH 6.5,inoculum size 10%,temperature 30℃,and cultivation time 25 h.

Objective To establish a general health information construction effect evaluation model for overall as-sessment of health information construction effect .Methods Domestic and foreign health information construction effect evaluation models were systematically analyzed by bibliometric analysis , comparative analysis , inductive and deductive method,repectively.The classic health information construction effect models were integrated.Results The health information construction effect evaluation model was established from the technique-organization man-agement-operation supportangle .Conclusion Thehealth information construction effects include technique effect, organization mangement effect, and operation effect.Thegeneral health information construction effect evaluation model is established, which includes 7 primary indexes and 20 secondaey indexes.

OBJECTIVETo elucidate the molecular background of Del phenotype in the Chinese population and explore new Del alleles.

METHODSFive hundred and fifteen RhD negative blood samples was tested by Rh typing test, indirect antiglobulin test and adsorption and elution assay to screen the Del phenotype. DNA of all the Del samples was analysed by multiplex polymerase chain reaction (MPX PCR) for the presence of RHD and by sequence-specific primer polymerase chain reaction (PCR-SSP) for Del alleles: RHD 1227A and RHD 885T. Samples which showed the negative result by PCR-SSP, were additionally analysed by genomic DNA sequencing and cDNA sequencing.

RESULTSSeventy-nine Del samples were found by adsorption and elution assay. All these samples had RHD exons 3, 4, 5, 6, 7 and 9. Except 4 Del samples, other 75 Del samples carried the RHD 1227A allele. None of the samples had the RHD 885T allele. Four novel RHD alleles were found in these four Del sample. There were RHD 3G-->A (GenBank DQ310735), RHD 28C-->T, RHD 53T-->C (GenBank DQ451877,DQ451878), RHD 251T-->C (GenBank DQ310734).

CONCLUSIONfnRh blood group system is very complex. New D variation phenotypes and new RHD alleles may be discovered ceaselessly.

Alleles ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; China ; Gene Frequency ; Genetics, Population ; methods ; Genotype ; Humans ; Molecular Sequence Data ; Mutation ; Phenotype ; Polymerase Chain Reaction ; Rh-Hr Blood-Group System ; genetics ; Sequence Analysis, DNA

OBJECTIVEDiagnosing neonatal asphyxia solely according to Apgar score may lead to misdiagnosis. The aim of this study was to explore new and more accurate diagnostic criteria for neonatal asphyxia.

METHODSTotally 10 376 live born neonates in our hospital were consecutively enrolled into the study. The following five items related to birth asphyxia, i.e., antepartum high-risk factors, Apgar scores, umbilical artery blood pH, organ injury, differential diagnosis on the causes of low Apgar score cases were examined and registered. The relationship among the first 4 items were analyzed. By differential diagnosis, the sensitivity and specificity of each index on diagnosing asphyxia and their complementary value on each other were investigated.

RESULTSThe items correlated well with each other (P < 0.01 or < 0.05) but were not entirely parallel and consistent; they could complement but could not substitute for each other. The sensitivity of antepartum high-risk factors, low Apgar scores, umbilical artery blood pH < 7.00 and organ injury was 100%, 100%, 44.44% and 100%, while the specificity was 17.99%, 98.90%, 96.05% and 96.62%, respectively. Of the 230 low Apgar score cases in this series only 50.9% coincided with asphyxia. For the 230 cases, when low Apgar score was combined with umbilical artery blood pH < 7.00, the sensitivity and specificity were 41% and 99.1% and when low Apgar score was combined with umbilical artery blood pH < 7.20, the sensitivity and specificity were 100% and 29.20%, respectively. After organ injury was added, the specificity was increased to 65.49%. When differential diagnosis was further added to exclude the other causes of low Apgar score cases, the misdiagnosis rate was minimized.

CONCLUSIONUp to now, no single accurate index for diagnosing neonatal asphyxia is available. In order to increase diagnostic bases and reduce misdiagnosis, the criteria of sole Apgar score should be replaced by multi-index diagnostic criteria. Based on the present study, a set of integrated diagnostic criteria for neonatal asphyxia is proposed: (1) prenatal high-risk factors, (2) low Apgar scores (respiratory depression must present), (3) umbilical artery blood pH < 7.00, if only pH < 7.20, the items (2) (4) (5) must be present, (4) hypoxic-ischemic organ injury (at least one organ dysfunction), (5) the other causes of low Apgar scores should be excluded. The last 4 indexes should all be met and the first one serves as reference. If multi-organ (three or more organs) dysfunction and (or) hypoxic-ischemic encephalopathy are present, severe asphyxia can be diagnosed.

Apgar Score ; Asphyxia Neonatorum ; blood ; diagnosis ; Diagnosis, Differential ; Diagnostic Errors ; prevention & control ; Humans ; Hydrogen-Ion Concentration ; Infant, Newborn ; Multiple Organ Failure ; Risk Factors ; Sensitivity and Specificity

OBJECTIVESTo determine the expression of nitric oxide synthase (NOS) in testis and to investigate the effects of NO on the reproductive function of testis.

METHODSTestes of adult male Sprague-Dawley rats were fixed in 4% paraformaldehyde. The paraffin sections were made as routine. Immunohistochemical ABC method was used to observe the localization of NOS.

RESULTSEndothelia NOS (eNOS), neuronal NOS (nNOS) and inductive NOS (iNOS) were all expressed in Leydig cells. Only eNOS was expressed in peritubular myoid cells, endothelial and smooth muscle cells of blood vessel, while only nNOS expressed in tunica adventitia of testicular blood vessels. The reactive substance distributes in cytoplasm with negative nuclei. Immunoreactivity for eNOS, nNOS and iNOS in all spermatogenic cells was negative.

CONCLUSIONSThree kinds of NOS were all expressed in testis and the distribution of different NOS had a little difference.

Animals ; Immunohistochemistry ; Male ; Nitric Oxide ; biosynthesis ; Nitric Oxide Synthase ; analysis ; Rats ; Rats, Sprague-Dawley ; Testis ; enzymology

OBJECTIVETo study the pathogenesis of abnormal bone metabolism in patients with HBV liver cirrhosis.

METHODSNM-300 signal-energy X-ray absorptiometry system was used to measure the bone mineral density (BMD) in 61 liver cirrhosis patients and 30 age-matched healthy controls. The serum levels of 1,25(OH)2D3, parathyroid hormone (PTH), calcitonin (CT), bone gamma-carboxyglutamic acid-containing protein (BGP), IL-1beta, IL-6, tumor necrosis factor (TNF)alpha and urine crosslaps were also detected in these patients.

RESULTSBMD in patients with HBV liver cirrhosis was lower than those of the controls. The serum levels of 1,25(OH)2D3 and BGP in cirrhosis patients were lower than those in the controls, and they were much lower in the osteoporosis (OP) group than in the non-osteoporosis (NOP) group. The PTH and CT were higher significantly in the patients than in the controls. The changes of serum 1,25(OH)2D3 and BGP were correlated with the changes of BMD. The serum levels of IL-1beta, IL-6, TNFalpha and urine crosslaps in cirrhosis patients were higher than those of the controls, and they were much higher in the OP group than in the NOP group. We also found that the serum levels of IL-1beta, IL-6, TNFalpha and urine crosslaps had a negative correlation with BMD.

CONCLUSIONSThese data suggest that bone formation is weakened and bone resorption is increased in patients with HBV liver cirrhosis, 1,25(OH)2D3 plays an important role in abnormal bone formation. Elevation of serum IL-1beta, IL-6, TNFalpha can accelerate bone resorption and cause hepatic bone disease (HBD). Taking 1,25(OH)2D3 and reducing the level of IL-1beta, IL-6, TNFalpha may be very important in preventing and treating HBD.

Adult ; Bone Density ; Bone and Bones ; metabolism ; Calcitriol ; pharmacology ; Hepatitis B, Chronic ; complications ; Humans ; Liver Cirrhosis ; complications ; Male ; Middle Aged ; Osteoporosis ; etiology

BACKGROUNDIn recent years, the prevalence of type 2 diabetes among Chinese population has been increasing by years, directly leading to an average annual growth rate of 19.90% of medical expenditure. Therefore, it is urgent to work on strategies to control the growth of medical expenditure on type 2 diabetes on the basis of the reality of China. Therefore, in this study, we explored the feasibility of implementing bundled payment in China through analyzing bundled payment standards of type 2 diabetes outpatient services.

METHODSThis study analyzed the outpatient expenditure on type 2 diabetes with Beijing Urban Employee's Basic Medical Insurance from 2010 to 2012. Based on the analysis of outpatient expenditure and its influential factors, we adopted decision tree approach to conduct a case-mix analysis. In the end, we built a case-mix model to calculate the standard expenditure and the upper limit of each combination.

RESULTSWe found that age, job status, and whether with complication were significant factors that influence outpatient expenditure for type 2 diabetes. Through the analysis of the decision tree, we used six variables (complication, age, diabetic foot, diabetic nephropathy, cardiac-cerebrovascular disease, and neuropathy) to group the cases, and obtained 11 case-mix groups.

CONCLUSIONSWe argued that it is feasible to implement bundled payment on type 2 diabetes outpatient services. Bundled payment is effective to control the increase of outpatient expenditure. Further improvements are needed for the implementation of bundled payment reimbursement standards, together with relevant policies and measures.

Aged ; Diabetes Mellitus, Type 2 ; economics ; Female ; Health Expenditures ; standards ; Humans ; Insurance, Health, Reimbursement ; Male ; Middle Aged ; Outpatients

Objective To investigate the effect of TiO2-SiO2 coatings with different firing temperatures to the Ti-porcelain bond strength.Methods The surface of titanium was coated uniformly with TiO2-SiO2 nano-coatings by solution-gelation (sol-gel) technology and then imposed three kinds of firing temperatures (300 ℃,500 ℃,750 ℃).The specimens without any coatings were the control group.The bond strength between the TiO2-SiO2 nano-coatings and titanium substrate was tested by the scarification.The Ti-porcelain bond strength was evaluated using three-point bending test.The phase composition of the TiO2-SiO2 nano-coatings was characterized by X-ray diffraction(XRD).The interface of Ti-porcelain and fractured titanium surface were investigated by scanning electron microscope (SEM).Results When the heat treatment temperature was 300 ℃,500 ℃ and 750 ℃,the bond strength of the coatings to the titanium substrate was (15.3±0.8),(22.1 ± 0.5),and (28.5 ±1.3) N respectively.The XRD patterns of the coatings showed that no rutile phase was found in all specimens.Compared with the Ti-porcelain system without any intermediate coatings [(29.5 ± 4.2) MPa],the bond strength of the same system coated with TiO2-SiO2 nano-coatings,being produced by the sol-gel technology and fired at 750 ℃ [(35.9 ± 2.3) MPa] was significantly increased by 21.8％.The SEM photomicrographs of titanium-porcelain interface morphology of the specimens before porcelain fused showed that the TiO2-SiO2 nano-coatings fired at 750 ℃ was compact and homogeneous without any obvious cracks.The location of the Ti-porcelain fracture existed within the TiO2-SiO2 nano-coatings.Conclusions TiO2-SiO2 nano-coatings by sol-gel technology fired at 750 ℃ can notablely improve the Ti-porcelain bond strength.

Objective To compare the effects of 20 mg qd and 10 mg bidadministration of iprrazole enteric-coated tablets on the control of gastric acid in healthy subjects.Methods A randomized,single-center,parallel controlled trial was designed to include 8 healthy subjects.Randomly divided into 2 groups,20 mg qd administration group:20 mg enteric-coated tablets of iprrazole in the morning;10 mg bid administration group:10 mg enteric-coated tablets of iprrazole in the morning and 10 mg in the evening.The pH values in the stomach of the subjects before and 24 h after administration were monitored by pH meter.The plasma concentration of iprazole after administration was determined by HPLC-MS/MS.The main pharmacokinetic parameters were calculated by Phoenix WinNonlin(V8.0)software.Results The PK parameters of iprrazole enteric-coated tablets and reference preparations in fasting group were as follows:The Cmax of 20 mg qd group and 10 mg bid group were(595.75±131.15)and(283.50±96.98)ng·mL-1;AUC0-t were(5 531.94±784.35)and(4 686.67±898.23)h·ng·mL-1;AUC0-∞ were(6 003.19±538.59)and(7 361.48±1 816.77)h·ng·mL-1,respectively.The mean time percentage of gastric pH＞3 after 20 mg qd and 10 mg bid were 82.64％and 61.92％,and the median gastric pH within 24 h were 6.25±1.49 and 3.53±2.05,respectively.The mean gastric pH values within 24 h were 5.71±1.36 and 4.23±1.45,respectively.The correlation analysis of pharmacokinetic/pharmacodynamics showed that there was no significant correlation between the peak concentration of drug in plasma and the inhibitory effect of acid.Conclusion Compared with the 20 mg qd group and the 10 mg bid group,the acid inhibition effect is better,the administration times are less,and the safety of the two administration regimes is good.

OBJECTIVETo study the possible mechanism of the intracellular aggregate formation of small heat shock protein HSPB8 (HSPB8)(K141N) mutation resulting in axonal Charcot-Marie-Tooth disease type 2L(CMT2L).

METHODSThe cell models which transiently expressed pEGFPN1-HSPB8 and pEGFPN1-(K141N)HSPB8 were established. The immunofluorescent co-location study of EGFP-(K141N)HSPB8 and HSPB1, EGFP-(K141N)HSPB8 and neurofilament light chain (NEFL) was carried out in the SHSY5Y cell models. The aggregate formation of EGFP-(K141N)HSPB8 in cell models was investigated and the possible mechanism of cellular aggregate formation was analyzed by t test and analysis of variance between group(ANOVA).

RESULTSEGFP-(K141N)HSPB8 formed large aggregate which predominantly located around the nucleus in cell models. EGFP-(K141N)HSPB8 co-localized perfectly with HSPB1 and NEFL in the SHSY5Y cell models. The aggregate formation was different in different cell types, there were fewer aggregates formed in an sHSPs deficient milieu than in HEK293T cells.

CONCLUSION(K141N)HSPB8 formed aggregates predominantly locate around the nucleus in cells. (K141N)HSPB8 co-localizes perfectly with HSPB1 and NEFL. The aggregate formation may be due to (K141N)HSPB8 conformational change leading to self aggregation and its abnormal interaction with other sHSPs such as HSPB1.

Cell Line ; Cell Line, Tumor ; Cell Nucleus ; metabolism ; Charcot-Marie-Tooth Disease ; genetics ; metabolism ; Green Fluorescent Proteins ; genetics ; metabolism ; HSP27 Heat-Shock Proteins ; HeLa Cells ; Heat-Shock Proteins ; genetics ; metabolism ; Humans ; Kidney ; cytology ; metabolism ; Microscopy, Confocal ; Neoplasm Proteins ; genetics ; metabolism ; Neuroblastoma ; genetics ; metabolism ; pathology ; Neurofilament Proteins ; genetics ; metabolism ; Point Mutation ; Protein-Serine-Threonine Kinases ; genetics ; metabolism ; Recombinant Fusion Proteins ; genetics ; metabolism ; Transfection