By functional plates,16 strains which can produce?-mannana-se were isolated frnm 28 Bacillus spp.Using a pair of degenerated primers,the conserved fragments of?-mannanase gene from the selected strains were amplified by PCR.The obtained nucleotide fragments were sequenced and compared with the homologous?-mannanase genes in GenBank and a phylogenetic tree was generated.Comparing to the genes coding?-mannanase published,the cloned nucleotide fragments show the highest sequence identity between 62% and 98%.The genes coding fnr?-mannanase of Bacillus circulus have low identity while the?-mannanase genes of Bacillus subtilis and other Bacillus spp. have high identity.

AIM:To be one of the primary cause injury to multiple sites of ocular of glaucoma which affects over 70 million people worldwide. We applied data mining techniques, linear and the matrix operations, efficiently calculated the network and estimated the possible function of the“node” genes of the retina and optic of glaucoma, in order to provide new thought and method on the pathogenesis of glaucoma.
METHODS: The data in this study is from Gene Expression Omnibus ( GEO ) which belong to Nation Center for Biotechnology Information ( NCBI) , the quality of the raw data CEL files was processed and analyzed by the Expression software which belong to Affymetrix Inc. , Santa Clare, CA, USA. Significant analysis method ( SAM) which base on the T test was used to identified the significant genes. Based on GRNInfer and Gvedit soft we set up gene networks of optic and retina of mice and further more enriched analysis which based on DAVID and MAS3. 0 online software were processed.
RESULTS:The analysis between the group of the optic nerve heads and retinas in different stage of glaucoma showed that the amount of significant different expressed genes in the optic never head group increased significantly comparing with the group of retina in the early stage of glaucoma, the analysis of the genes network construction show that:the node genes of optic nerve heads included Unc13c、Kif5a、TRPM1、PANX; and the node genes of retina include POU4F1, NEFL, BC03870, CALB2. Metabolic pathways enrichment analysis which based on MAS3. 0 online platform show that there was mainly the amyotrophic lateral sclerosis, tyrosine metabolism, melanogenesis, Nitrogen metabolism, Gap junction, Leukocyte transendothelial migration metabolism pathway enriched out in optic nerve head; and there was mainly amyotrophic lateral sclerosis, neurodegenerative disorders, prostate cancer, leukocyte transendothelial migration metabolism pathway enriched out in retina.
CONCLUSION:By understanding bioinformatics result, it seems optic were more sensitive than the retina to high intraocular pressure, and weather high expression of TYrp1 gene can be as a sensitive diagnostic item require more evidence back up. Functional enrich analysis of node gene showed that cytoskeleton reconstructed, molecular motor and nutrients transport function improve in optic; and in retina, the most prominent finding in retina was enrichment function modules were focus on regeneration, repairing and differentiation of cells, which remind that we should reinforce research on reparation of retina of primary glaucoma. Metabolic pathways enrichment analysis show that inflammatory response plays prominent place in optic and retina of primary glaucoma, because of the optic narrow and crowed anatomic shape, nutrient metabolism and substances transfer enrichment modules play an important role in optics of primary glaucoma.

Objective To investigate the curative effect of brucellar spondylitis,so as to provide scientific proof for improving the curative level of the disease.Methods Epidemiological information was collected from 113 patients diagnosed as brucellar spondylitis,who were divided into 5 groups according to different drugs and drug combinations of doxycycline,gentamicin,sulfamethoxazole and rifampicin.Then the curative effect was investigated.Twenty-one patients who had greater paoas muscle abscess or Para vertebral abscess,intraspinal abscess of spinal canal,necrotic intervertebral disk and major osteolasia received the minimal invasive surgery and the focus removal surgery.Results The occurrence of the disease in female was much higher than that in male.Grazing and breeding beasts was the principal route of infection.Lumbars was mostly involved.they usually was infected in the adjacent 2 piece.L4 was the most common and seriuous one.The curative effect of doxycycline group was better than that without doxycycline(72.60% vs 35.00%,X2=15.14,P＜0.05).Doxycyeline+gentamicin+sulfamethoxazole was reeommended as the first choice.However,the curative effect did not increase despi~course of the treatment prolonged.The heMing rate and effective rate after 1 course was 52.21%(59/113)and 92.04%(104/113).that after 2 courses 58.41%(66/113)and 95.58%(108/113),that after 3 courses 59.29%(67/113)and 95.58%(108/113).The healing rate in different course did not present differences(P＞0.05).21 patients undergoing surgery were followed-up,12 patients were after 2 years and 9 patients were between 1-2 years.The healing rate was 95.24%(20/21),1 case was healed basically,the effective rate was 100%.None reoccurred.Conclusions There are characterized features in clinical epidemiology of the brucell spondylitis.Long term,adequate in dosage,combination and multi-approach use of antibiotics is the most reliable way to treat and prevent it from recurring.But fof the patients soitable for surgery.the minimal invasive surgery or the focus removal could shorten the course of therapy,decrease the complications and increase the cure rate.

The related substances in docetaxel injection were identified by LC-MS/MS. Ethyl acetate was used to extract the injection to remove the pharmaceutical excipients. HPLC separation was carried out on a Hedera ODS-2 column (150 mm x 2.1 mm, 5 microm) with a mobile phase consisting of acetonitrile - 0.1% acetate acid aqueous solution (40: 60). Electrospray ionization source was set in the positive mode for the LC-ESI-MS/MS, and the ion monitoring modes were full scan and product ion scan. According to the mass spectra of the related substances, the fragment profiles were explained, and the chemical structures were elucidated. Docetaxel and its main related substances were well separated. Nine related substances in docetaxel injection were detected by LC-MS/MS. Their chemical structures were proposed, and four of them were identified in the docetaxel injection for the first time. The established LC-MS/MS method is effective in the separation and identification of the related substances in docetaxel injection. The test results are useful for its quality control.
Antineoplastic Agents ; administration & dosage ; analysis ; chemistry ; Chromatography, High Pressure Liquid ; Drug Contamination ; Injections ; Molecular Weight ; Quality Control ; Spectrometry, Mass, Electrospray Ionization ; Tandem Mass Spectrometry ; Taxoids ; administration & dosage ; analysis ; chemistry

OBJECTIVETo observe clinical characteristics of herb-induced liver injury (HILI).

METHODSGeneral conditions, medical history, clinical manifestations, biochemical indices, prognosis, and Roussed Uclaf Causality Assessment Method (RUCAM) scores were retrospectively analyzed in 595 inpatients at 302 Military Hospital between January 2009 and January 2014.

RESULTSThere were 423 cases (accounting for 71.1%) were females with multiple onset age ranging 41 to 50 years old. The median time from starting Chinese herbs to the occurrence of liver injury (LI) was 30 days (15-75 days), and 511 cases (85.9%) were classified as hepatocellular injury. Chinese herbs inducing HILI were mainly used for skin disease (102 cases, 17.1%), osteoarticular disease (57 cases, 9.6%), and gastrointestinal disease (49 cases, 8.2%), covering 207 kinds of Chinese patent medicines. Polygonum multiflorum, Psoralea corylifolia, and Corydalis ambigua were often seen in Chinese prescriptions. In RUCAM scoring, 451 HILI patients (accounting for 74.1%) were very possibly associated with Chinese herbs. Liver failure occurred in 47 HILI patients (accounting for 7.9%), cirrhosis in 45 patients (accounting for 7.6%), chronic HILI in 80 patients (accounting for 13.4%), 27 (4.5%) died, and only 2 (0.3%) underwent liver transplantation.

CONCLUSIONSChinese herbs could cause LI or even death. Attention should be paid to herbal hepatotoxicity and improving monitoring system of HILI.

Chemical and Drug Induced Liver Injury ; diagnosis ; Female ; Humans ; Prognosis ; Retrospective Studies

Decoction is one of the most commonly used dosage forms of traditional Chinese medicine. The stability of chemical constituents in decoction is closely related to the clinical efficacy and safety. There were few reports about the influence of metal ions in the stability of chemical constituents in traditional Chinese medicine. However, there is no evidence that metal ions in decoction water need to be controlled. In this study, 2,3,5,4'-tetrahydroxy stilbene-2-O-β-D-glucoside (THSG), one of the main constituents in Polygoni Multiflori Radix was studied. Ordinary tap water, deionized water, and water containing different metal ions were used to investigate and compare the influence on THSG. The results showed that after storage in a dark place at the room temperature for 10 days, the degradation of THSG was 7% in deionized water, while undetectable in tap water. The content of THSG could be decreased by different kinds of metal ions, and the effect was concentration-dependent. Moreover, Fe3+ and Fe2+ showed the greatest influence at the same concentration; and our study has shown that THSG decreased more than 98% in Fe and Fe2+ solutions at 500 ppm concentration. In the same time we found out p-hydroxybenzaldehyde (molecular weight: 122.036 7) and 2,3,5-trihydroxybenzaldehyde-2-O-glycoside (molecular weight: 316.079 4) were the main degradation products of THSG in tap water and water containing Cu2+, Ca2+, Zn2+, Mg2+ and Al3+. The product of THSG dimer with a water molecule was found in water containing Fe3+ and Fe2+. The above results showed that the metal ions in water could significantly influence the stability of THSG in water, indicating that the clinical efficacy and safety of decoction would be affected if the metal ions in water were not under control. It's suggested that deionized water should be used in the preparation of decoction containing Polygoni Multiflori Radix in the clinic to avoid degradation of THSG. Meanwhile, decoction prepared by tap water should be taken by patients in a short time. Our investigation provides important information and reference about the influence of metal ions on the stability of decoctions in other traditional Chinese medicine that have unstable groups such as hydroxyls and unsaturated bonds, etc.
Drugs, Chinese Herbal ; chemistry ; Glucosides ; chemistry ; Ions ; chemistry ; Metals ; chemistry ; Plant Roots ; chemistry ; Polygonaceae ; chemistry ; Stilbenes ; chemistry

The hybrid xylanase TB was constructed by the substitution of the N-terminus segment of the Streptomyces olivaceoviridis xylanase XYNB with corresponding region of Thermomonosporafusca xylanase TfxA. The hybrid gene tb, encoding the TB, was correctly expressed in Escherichia coli BL21 and Pichia pastoris GS115. TB was purified and its enzymatic properties were determined. The results revealed that the optimal temperature and optimal pH of TB were at 70 degrees C and 6.0, which have been obviously improved compared with those of XYNB. The thermostability of TB were all about six-fold of XYNB's after incubating the properly diluted enzyme solutions at 80 degrees C and 90 degrees C for 3min, respectively. The pH stability of TB was 5 to approximately 9, which was narrower than that of XYNB. Still, TB remains a high specific activity as XYNB does. Analysis of a homology modeling and sequence similarity were used to reveal the factors influencing the enzymatic properties of TB and the discussion for the relationship between structure and function of xylanase was given.
Amino Acid Sequence ; Base Sequence ; Desulfurococcaceae ; enzymology ; genetics ; Endo-1,4-beta Xylanases ; genetics ; metabolism ; Enzyme Stability ; Escherichia coli ; enzymology ; genetics ; Hot Temperature ; Molecular Sequence Data ; Pichia ; enzymology ; genetics ; Protein Engineering ; methods ; Recombinant Fusion Proteins ; genetics ; metabolism ; Streptomyces ; enzymology ; genetics ; Structure-Activity Relationship

A gene appA encoding a novel phytase was firstly cloned from Hafnia alvei by PCR and sequenced. The gene was consisted of 1335 bp, encoding 444 amino acids. The calculated molecular weight of the mature APPA was about 45.2 kD. The gene appA was expressed in E. coli BL21 (DE3). Recombinant APPA was purified and its enzymatic properties were determined. The optimum pH for the enzyme was 4.5 and the optimum temperature was 60 degrees C. The pH stability of r-APPA is good, the relative phytase activity was above 80% after treated in buffers of pH 2.0-10.0. The specific activity of r-APPA is 356.7 U/mg, and the Km value was 0.49 mmol/L and Vmax of 238 U/mg. The enzyme showed resistance to pepsin and trypsin treatment.
6-Phytase ; biosynthesis ; genetics ; isolation & purification ; Amino Acid Sequence ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Hafnia ; enzymology ; genetics ; Hydrogen-Ion Concentration ; Molecular Sequence Data ; Recombinant Proteins ; biosynthesis ; genetics ; isolation & purification ; metabolism ; Temperature

Aspergillus fumigatus wild-type phytase has many favorable properties, such as a good thermorstability and a broad pH optimum. However, the specific activity of the enzyme is relative low. A. fumigatus Q23L phytase resulted in a remarkable increase in specific activity around pH4.5 - 7.0, but the pH stability of Q23L was lower than A. fumigatus wild-type phytase. To increase the pH stability of Q23L, the mutant Q23LG272E was constructed by site-directed mutagenesis with PCR. The gene of A. fumigatus wild-type phytase and the mutant genes encoding the Q23LG272E and the Q23L were correctly expressed in Pichia pastoris GS115. Enzymes were purified and their enzymatic properties were determined. The results revealed that the specific activity of the Q23L improved remarkably, which increased from 51 u/mg of the wild type to 109 u/mg at pH5.5. Meanwhile, the pH stability of Q23L, decreased evidently, especially from pH3.0 to pH4.0.The pH stability of Q23LG272E in pH3.0 - 4.5 and pH6.5 - 7.0 has been improved compared with Q23L. The specific activity of Q23LG272E basically maintained at the level of Q23L. Analysis of 3-D structure and sequence similarity were used to reveal the presumable factors influencing the enzymatic properties of Q23LG272E, and discussion for the relationship between structure and function of phytase was given.
6-Phytase ; chemistry ; genetics ; metabolism ; Amino Acid Substitution ; Aspergillus fumigatus ; enzymology ; genetics ; Biocatalysis ; Electrophoresis, Polyacrylamide Gel ; Fungal Proteins ; chemistry ; genetics ; metabolism ; Hydrogen-Ion Concentration ; Models, Molecular ; Mutagenesis, Site-Directed ; Mutant Proteins ; genetics ; metabolism ; Mutation ; Pichia ; genetics ; Polymerase Chain Reaction ; Protein Conformation ; Protein Engineering ; methods ; Recombinant Proteins ; metabolism ; Structure-Activity Relationship ; Substrate Specificity

OBJECTIVETo evaluate the feasibility and the clinical practicality of the design about the hollowed titanium stick supporting the femoral head and preventing it from collapsing.

METHODSFrom Jan.2003 to Jun.2007, 36 patients (46 hips) diagnosed as cystis degeneration of the femoral head were treated by surgical operation, including 20 males and 16 females with an average age of 40 years ranging from 18 to 56 years old, and the course of the disease was from 10 to 24 months (16 months on average). According to ARCO staging,there were 24 patients (34 hips) in NFH II, of which 11 hips were II a stage, 13 hips were II b stage and 10 hips were II c stage; there were 6 patients diagnosed as osteofibrous,4 patients as simple bone cyst and 2 patients as chondromyxoid fibroma. Under X-ray the percutaneous narrow core decompression and focus infection elimination were performed and supported the sclerotin under the cartilage with titanium stick. The patients were followed-up at the first, third, sixth, twelfth, twenty-fourth and thirty-sixth month after the operation. The clinical evaluation was done by X-ray and the indexes included stable, unstable and abortive. The data was analyzed by Fisher exact probility and the suviaval rate was analyzed by Kaplan-Meier suviaval curve using statistical soft ware SPSS13.5.

RESULTSThere were no unstable or failure cases on each period from the 1st month to the 12th month after the operation, indicating that the supporting effect of the titanium stick was exact during 12 months after the operation. There were unstable and failure cases from the 12th month to the 24th month after the operation, which were mainly in stage NFH II c but the comparision of the stable rate in this period and 12 months after the operation had no obvious statistical differences (P>0.05) indicating that the supporting effect of the titanium stick was feasible during the stage. One of the unstable cases deteriorated and failed but there were no new unstable cases, both the stable rate and the unstable rate had no change and the failure rate rose on the 36th month after the operation. The compar- ision of the stable rate on each period after the operation had no obvious difference (P=0.197>0.05), which indicated that the supporting effect of the titanium stick was persistent. By the difference of the etiology the three-year survival rate of the relevant NFH II c pathological changes was the lowest-70% and the survival rate of the pathological changes induced by other etiological factors was 90.2%.

CONCLUSIONThe design about the hollowed titanium stick supporting the collapsed femoral head is feasible. Using the hollowed titanium stick to support the femoral head and prevent it from collapsing is pragmatic in the clinical and the effect is positive, however, when it comes to the NFH II c pathological changes, the choice should be made discreetly.

Adolescent ; Adult ; Feasibility Studies ; Female ; Femur Head ; diagnostic imaging ; surgery ; Follow-Up Studies ; Humans ; Internal Fixators ; adverse effects ; Male ; Middle Aged ; Prosthesis Design ; methods ; Titanium ; Tomography, X-Ray Computed ; Young Adult