Objective: To observe the histomorphology/pathology changes and the expression of hypoxia inducible factor-1 alpha (HIF-1?) and vascular endothelial growth factor (VEGF) in drowning rats brain, in an attempt to explore new pathways for treatment of water drowning. Methods: The animal model of freshwater drowning was established with rats. The rats were randomly divided into the following groups: control group, drowning-to-death group and 2 min-drowning group (including 5 subgroups survived for 3, 6, 12, 24. and 40 hours after being drowned in water for 2 min [n=10]). Brain indices of rats in each group were detected and the pathological changes of cerebral tissues were observed; the expression of HIF-lo and VEGF were detected immunohistochemically and the correlation between them was analyzed. The dynamic blood oxygen saturation was detected in rats survived for 40 h after 2 min-drowning at different points. Results: Brain index of the drowning group and groups surviving for 24, 40 hours after drowning were obviously higher than that of the control group (P

Brain natriuretic peptide (BNP) is one of the most sensitive and specific laboratory indicators during cardiac dysfunction.N-Terminal pro-brain natriuretic peptide (NT-proBNP) is the N-terminal precursor of BNP.The functions of both are not consistent,but its physicochemical properties are superior to BNP.Now it has been widely used in the diagnosis and treatment monitoring of cardiovascular diseases.In the field of stroke,the testing and application of BNP,especially NT-proBNP is less.This article reviews the roles of NT-proBNP in cardio-cerebrovascular diseases,especially in ischemic cerebrovascular disease.

The aim was to establish an effective screening microarray at genus level for Pospiviroid. We analyzed nucleotide sequences from Pospiviroid viroid and designed 19 probes with genus identification characteristics. The standards of these probes included the characters of (i) a GC content between 40 and 60%, (ii) less than 50% of single nucleotide, (iii) less than 4 continuous mononucleotides, and (iv) less than 6 nucleotides in the inner hairpin. We synthesized microarrays by using these probes on glass slides. The validation results of microarray probes show effective signals from chrysanthemum stunt viroid and tomato planta macho viroid standard samples hybridization. The sensitivity results show that the microarray detected 200 pg/microL of total RNA. The microarray can be used to screen Pospiviroid viroid.
Base Composition ; Base Sequence ; Microarray Analysis ; Nucleic Acid Hybridization ; Plant Diseases ; virology ; Plant Viruses ; classification ; RNA ; Viroids ; classification

To express feruloyl esterase A from Aspergillus oryzae in Pichia pastoris expression system and study its hydrolysis function, explore the conditions and effects of purification for ferulic acid extracts by macroporos resin. Using the total RNA from A. oryzae CICC 40186 as the template, we amplified coding sequence AorfaeA encoding a mature feruloyl esterase A (AorFaeA) by RT-PCR technique. Then, the coding sequence AorfaeA was successfully expressed in Pichia pastoris GS115 mediated by an expression plasmid pPIC9K. The purified recombinant AorFaeA (reAorFaeA) showed one single band on SDS-PAGE with an apparent molecular weight of 39.0 kDa. The maximum activity of reAorFaeA to methyl ferulate, measured by high-performance liquid chromatography (HPLC), was 58.35 U/mg. Then, reAorFaeA was used to release ferulic acid from de-starched wheat bran in the presence of xylanase. The purification tests for ferulic acid from the enzymatic hydrolysate were carried out with preselected macroporous resins. The results showed that macroporous resin HPD-300 had much higher adsorption and desorption capacities. Ferulic acid could be quantitatively recovered by 50% of the eluent concentration at a flow speed of 1 mL/min. Under the purification condition, the recovery ratio of ferulic acid was 92%, and the content of ferulic acid was increased from 0.13% in the raw material to 10.55%. This work exploits the breakdown of ferulic acid by recombinant enzymeand provids a good strategy to its "green production".
Aspergillus oryzae ; enzymology ; Carboxylic Ester Hydrolases ; biosynthesis ; genetics ; Cloning, Molecular ; Coumaric Acids ; chemistry ; Electrophoresis, Polyacrylamide Gel ; Hydrolysis ; Molecular Weight ; Pichia ; genetics ; metabolism

Objective To investigate the effects of compound cornu cervi degelatinatum on the expression of related inflammatory cytokines and pathological changes of the synovium in mice with collagen-induced arthritis (CIA). MethodsThe CIA model of Balb/c mice was established. The mice were randomly divided into 5 groups: blank control group, model control group, low-dose group ( C1 ) , high-dose group ( C2 ) and glucosidorum tripterygll totorum group D. After the first immunization in mice, we observed the general condition and lesions of the fore and hind jaws. After the second immunization, the mice in groups C1 and C2 were orally administered with compound cornu cervi degelatinatum at the doses of 2. 5 and 5 g·kg-1 body weight per day for 4 weeks, and meanwhile those in group D were administrated with glucosidorum tripterygll totorum at the dose of 13. 6 mg·kg-1 body weight per day. The serum of the mice was collected to detect the levels of TNF-αand IL-4 by ELISA. Ankle joints were harvested, and the pathological changes of synovial tissues were observed under light microscope. Results As compared with the model control group, the level of TNF-αin the treatment groups was significantly decreased, while the level of IL-4 was elevated in group C1. Histological pathology of ankle joints demonstrated that the synovium of the CIA mice were hyperplastic and the synovial tissues were markedly ameliorated in treatment groups. Conclusion The compound cornu cervi degelatinatum can relieve redness and swelling in mice with CIA. The mechanism may be related to the inhibition of pro-inflammatory cytokine TNF-α and regulation of anti-inflammatory cytokine IL-4.

Objective To evaluate the comprehensive clinical ability of new nurses in hospital of traditional Chinese medicine by the application of obstructive structured clinical examination.Methods New nurses of Nanjing hospital of traditional Chinese medicine (TCM) of the grade 2012 and 2013 were assessed.The examination consisted of four parts including standard patient station,the capacity of the case,the basic nursing skills of medicine and TCM.The scores of the theoretical knowledges and nursing skills were assessed.Results The average score of the nurses was (76.32±6.31)points,complying to the normal distribution;The degree of difficulty of the examination was 0.70-0.85 and the degree of distinguish was 0.28-0.48;The score of the theoretical test was lower than that of nursing skills and there was significant difference between them (P＜0.05).Conclusions The design of the examination is reasonable and the clinical ability of new nurses majored in TCM can be well assessed.

Objective To investigate the application of three stages enhanced scan of multi-slice spiral CT for the characteristic analysis on the wall and lumen of colorectal mucous adenocarcinoma.Methods 51 patients with colorectal mucinous adenocarcinoma who had complete pathologic diagnosis were studied.GE Discovery CT 750HD scanner was used for line scan,arterial and portal venous phase scan,and delayed scan.The lesions of intestinal wall,enteric cavity and proximal normal lumen were observed by line scan and enhanced scan,and the related quantitative value and CT value were observed.Results The intestinal wall was uneven annular thickening or hemispherical thickening.The thickening intestinal wall showed lobular changes around the inner margin,outer margin,or without lobular changes.The hierarchical sign and cystoids sign were observed with three layers or two layers structure after enhanced scan.Metastases of the larger lymph node,liver,ovaries were characterized similar to cyst.According to the form,the stenosis was divided into natural streamlined,irregular fixed,and irregular constrictive types.The proximal normal intestinal canal of the lesion showed occlusion,mild expansion or severe expansion.Conclusions Walls of colorectal mucous adenocarcinoma was uneven annular thickening or hemispherical thickening,in which calcification was found,and there was features of similar to layer and cyst after enhancement.The stiff intestinal wall appeared less,and the proximal bowel obstruction was rare.Therefore,depend on these characteristics,the diagnosis of colorectal mucous adenocarcinoma can be achieved.

Objective:To determine the valproate concentration in plasma of epilepsy patients by HPLC, and compare with the re-sults of chemiluminescence microparticle immunoassay ( CMIA) to evaluate the consistency of the two methods. Methods:HPLC and CMIA was respectively applied to determine the plasma concentration of valproate in 230 epileptic patients. The correlation of the two methods was studied by Passing-Bablok regression and Bland-Altman method. Results:The regression equation of the determination re-sults of HPLC (Y) and CMIA (X) was Y=1. 069 7X+2. 338 2 (R2 =0. 969, n=230), which showed promising correlation. Bland-Altman analysis showed that the consistency of the two methods was poor, and the values of HPLC were higher. Conclusion: HPLC and CMIA used for the determination of valproate plasma concentration show good correlation. However, the consistency is poor and there is system error. In the clinical treatment, adjustment and choice should be paid more attention.

Objective To prepare endo-beta-N-acetylglucosaminidase H (Endo-H) expressed in Pichia pastoris, and apply it to N-glycosylation analysis .Methods One complete gene was synthesized on the basis of the cDNA sequence encoding Streptomyces plicatus reported in GenBank .The gene was cloned into the expression vector pPIC 9.The expression vector pPIC9-Endo-H was transformed into P.pastoris(JC308).The expression products were induced by methanol , puri-fied by two-step chromatography , used to analyze the glycan structures of RNaseB by the DNA sequencer assisted fluoro-phore-assisted carbohydrate electrophoresis (DSA-FACE)methods, and finally compared with peptide-N-asparagine amidase F(PNGase F).Results This enzyme expressed in P.pastoris(JC308) had the ability to hydrolyze natural or denatured high-mannose type of oligosaccharide linked by β-1,4-glycosidic bonds , but not complex-type oligosaccharide .The result of DSA-FACE showed that carbohydrate chains of Man 5 GlcNAc-Man9 GlcNAc could be obtained when RNaseB was hydrolyzed by Endo-H, and that Man5 GlcNAc2-Man9 GlcNAc2 chains became available when RNaseB was hydrolyzed by PNGase F . Conclusion Endo-H expressed in P.pastoris has bioactivity which can be used to analyze N-glycosylation with the method of DSA-FACE.

ObjectiveTo investigate the role played by acute glucagon response in the short-term intensive insulin therapy induced long-term remission of newly-diagnosed type 2 diabetes.MethodsTen newly-diagnosed type 2 diabetic patients( 7 males,3 females) received intensive continuous subcutaneous insulin infusion( CSII )therapy for 2 weeks.Intravenous glucose tolerance test (IVGTT) and hyperinsulinemia euglycemia clamp test were performed before and after CSII.Glucose infusion rate ( GIR ),acute insulin response ( AIR ) and acute glucagon response (AGR) were assessed.Long-term remission was defined as good glycaemic control without any hypoglycaemic agent (fasting plasma glucose ＜ 7.0 mmol/L 及 2 h postprandial glucose ＜ 10 mmol/L) one year after CSII.Relationship between AGR and long-term remission were analyzed by one way ANOVA and Spearman correlation analysis.Results ( 1 ) Five of the ten patients achieved long-term remission in this study.GIR and AIR in the remission group were increased significantly after CSII compared with those before CSII [ GIR ( 5.39 ± 1.76 vs 2.02 ± 0.83 )mg · kg-1 ·min-1,AIR ( mean 54.1 vs mean 3.39 ) mU · L-1 · min,P＜0.01 respectively ],however both of them were not associated with the remission.(2) AGR in the remission group was significantly higher than that in the non-remission group before CSII ( 5.10±0.60 vs 2.85 ± 1.86,P＜0.05 ) and was decreased significantly after CSII.The mean of AGR after CSII was apparently lower in the remission group than that in non-remission group (0 vs 3.04±2.00,P＜0.01 ).(3)Spearman analysis showed that AGR before CSII and its range of reduction after CSII were correlated with remission ( r for both were 0.731,P=0.016).ConclusionHigher level of AGR before CSII and greater reduction after CSII in the subjects with newly-diagnosed type 2 diabetes were significantly associated with long-term remission,suggesting that pancreatic α cells may play a unique role in the induction of remission of type 2 diabetes.