OBJECTIVETo promote development and utilization of Ophiocordyceps gracilis in xinjiang and provide basic data for researching and sustainable developing medicine fungus related to O. gracilis.

METHODA white strain SFYT002 isolated from the sclerotium of O. gracilis in Xinjiang was researched by morphological observation, ITS and 18SrDNA sequencing. The ITS and 18SrDNA sequences of the strain were determined, BLAST was compared with the other sequences of Tolypocladium in GenBank. The phylogenetic trees of ITS and 18SrDNA sequences were analyzed in Tolypocladium. In addition, the filter paper method was used to study the antibacterial effects.

RESULTThe main morphological characters of this strain were white cotton-like colonies, phialide with inflated base, drastically sharping with partially bending tips, small and transparent budding spores with being always assemble to spearhead and globular, subglobular or ellipse conidiospores. The phylogenetic trees of ITS and 18SrDNA sequences were constructed and analyzed in Tolypocladium. It was resulted that Tolypocladium was confirmed to be monophyletic, and the strain SFYT002 was the same as the systematic position of others of T. inflatum. Meanwhile, the antibacterial test was performed against the 4 common pathogenic bacteria. It was showed that both fermentation and its extracts of different polar from this strain possessed good anti-bacteria capacities.

CONCLUSIONThe strain SFYT02 was identified as T. inflatum, and inhibited effectively growth of bacteria.

Anti-Bacterial Agents ; isolation & purification ; pharmacology ; China ; DNA, Fungal ; genetics ; DNA, Intergenic ; genetics ; Hypocreales ; genetics ; isolation & purification ; physiology ; Medicine, Chinese Traditional ; methods ; Mycelium ; Phylogeny

So far there exists no corresponding quality test procedures and grading standards for the seed of Prunus humilis, which is one of the important source of base of semen pruni. Therefor we set up test procedures that are adapt to characteristics of the P. humilis seed through the study of the test of sampling, seed purity, thousand-grain weight, seed moisture, seed viability and germination percentage. 50 cases of seed specimens of P. humilis tested. The related data were analyzed by cluster analysis. Through this research, the seed quality test procedure was developed, and the seed quality grading standard was formulated. The seed quality of each grade should meet the following requirements: for first grade seeds, germination percentage ≥ 68%, thousand-grain weight 383 g, purity ≥ 93%, seed moisture ≤ 5%; for second grade seeds, germination percentage ≥ 26%, thousand-grain weight ≥ 266 g, purity ≥ 73%, seed moisture ≤9%; for third grade seeds, germination percentage ≥ 10%, purity ≥ 50%, thousand-grain weight ≥ 08 g, seed moisture ≤ 13%.
Cluster Analysis ; Germination ; Prunus ; growth & development ; Seeds ; physiology

ObjectiveTo introduction of perforator flaps,muscle flaps pedicled by descending branch of lateral circumflex femoral artery,method and their clinical application that multiple soft tissue defects of hand are repaire by muliplefoliated tissue flap only branch lateral circumflex femoral artery.MethodsFifteen patients with multiple soft tissue defects of hand were repaired muliplefoliated tissue flap only pedicled bydescending branch lateral circumflex femoral artery.At first,the anterolateral thigh perforator flap was designed and harvested according to the soft tissue defects of hand, then the descending branch lateral circumflex femoral artery was dissected at the same time the segmented perforator flap,fascia lata flap,rectus femoris muscle flap, vastus lateralis muscle flap, vastus intermedius muscle flap and distal spatium intermusculare flap were harvested in need according to distance among soft tissue defects.The muliplefoliated tissue flap was harvested only pedicled by descending branch lateral circumflex femoral artery, at last muscle flaps and fascia lata flaps were covered by skin graft, so the multiple soft tissue defects of hand were repaired in one time.ResultsNo vascular crisis happened. All skin grafts survived well, the contour of all repaired soft tissue defects was good and protective feeling was recovered by skin grafts of all flaps. All cases were got follow-up and the range was from 6 to 20 months(the average was 8.7 months).Wound of donor site healed well, muscle strength of quadriceps and motion of knee were normal. Three cases were excellent,nine cases were well and 3 cases were good, according to upper extremity function evaluation criteria of Chinese Medical Society for the Surgery of the Hand, the rate of good was 80 percent.ConclusionMultiple soft tissue defects of hand can be repaired by muliplefoliated flap only pedicled by descending branch of lateral circumflex femoral artery. Its advantages included reduction of operation time and treatment, good recovery of hand contour and function. It is a good method to repair multiple soft tissue defects of hand.

Acute Disease ; Animals ; Bone Marrow Cells ; cytology ; Cell Differentiation ; Female ; Liver Diseases ; pathology ; Male ; Mesenchymal Stromal Cells ; cytology ; Rats ; Rats, Sprague-Dawley

This study was designed to use iTRAQ technology coupled with 2D LC-MS/MS to study the comparative proteomics of different processing technology for pilose antler. 1015 proteins were identified with 2D LC combined with MOLDI TOF/TOF mass spectrometry. Comparative analysis with Protein Pilot (Version 4.5) revealed that 87 proteins were changed (P ≤ 0.05, the ratio of > 1.50 or < 0.60 as the threshold selection of difference proteins), of which 24 were up regulated and 33 were down regulated in the traditional frying process (TFP) compared with the fresh pilose antler (P ≤ 0.05). 7 significant different proteins (P ≤ 0.001), most of these significantly changed proteins were found to be involved in calcium ion binding and ATP binding associated with human healthy. Freeze drying with protective agent (FDP) (Trehalose) can improve the content of significantly different proteins (P ≤ 0.001) including Collagen alpha-1 (XII) chain (COL12A1) and Collagen alpha-1 (II) chain (COL2A1). The significant function involves in platelets activating, maintenance of spermatogonium, and disorder expression in tumor cells. The functional annotation by Hierarchical clustering and GO (gene ontology) showed that the main molecule functions of the proteins significantly changed in these processes were involved in binding (52.7%), catalytic (25.3%), structural molecule and transporter (6.6%).
Animals ; Antlers ; chemistry ; Chromatography, Liquid ; Collagen ; chemistry ; Down-Regulation ; Freeze Drying ; Gene Expression Regulation ; Proteomics ; Tandem Mass Spectrometry ; Technology, Pharmaceutical ; methods ; Up-Regulation

OBJECTIVETo construct lipL32/1-lipL41/1 fusion gene and its prokaryotic expression system and to determine frequencies of carrying and expression of lipL32 and lipL41 genes in L.interrogans wild strains and specific antibody levels in sera from leptospirosis patients.

METHODSlipL32/1-lipL41/1 fusion gene was constructed using linking primer PCR method and the prokaryotic expression system of the fusion gene done with routine techniques. SDS-PAGE was used to examine expression of the target recombinant protein rLipL32/1-rLipL41/1. Immunogenicity of rLipL32/1-rLipL41/1 was identified by Western blot. PCR and MAT were performed to detect carrying and expression of lipL32 and lipL41 genes in 97 wild L.interrogans strains. Antibodies against products of lipL32 and lipL41 genes in serum samples from 228 leptospirosis patients were detected by ELISA method.

RESULTSThe homogeneity of nucleotide and putative amino acid sequence of lipL32/1-lipL41/1 fusion gene were 99.9 % and 99.8 % in comparison with the reported sequences. Expression output of the target recombinant protein rLipL32/1-rLipL41/1, mainly present in inclusion body, accounted for 10 % of the total bacterial proteins. Both the rabbit antisera against rLipL32/1 and rLipL41/1 could combine to rLipL32/1-rLipL41/1. 97.9 % and 87.6 % of the L.interrogans wild strains had lipL32 and lipL41 genes, respectively. 95.9 % and 84.5 % of the wild strains were positive for MAT with titers of 1:4 - 1:128 using rabbit anti-rLipL32s or anti-rLipL41s sera, respectively. 94.7 % - 97.4 % of the patients'serum samples were positive for rLipL32s antibodies, while 78.5 % - 84.6 % of them were rLipL41s antibodies detectable.

CONCLUSIONlipL32/1-jlipL41/1 fusion gene and its prokaryotic expression system were successfully constructed. The expressed fusion protein had qualified immunogenicity. Both the lipL32 and lipL41 genes are extensively carried and frequently expressed by different serogroups of L.interrogans, and their expression products exhibit cross-antigenicity.

Antibodies, Bacterial ; blood ; Antigens, Bacterial ; genetics ; immunology ; Bacterial Outer Membrane Proteins ; genetics ; Cloning, Molecular ; Enzyme-Linked Immunosorbent Assay ; Gene Expression Regulation, Bacterial ; Genes, Bacterial ; genetics ; Humans ; Leptospira interrogans ; genetics ; Leptospirosis ; immunology ; microbiology ; Lipoproteins ; genetics ; Prokaryotic Cells ; metabolism ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology

OBJECTIVESTo investigate mechanism for the increasing level of serum vascular endothelial growth factor(VEGF) in tumour patients during radiotherapy and the inhibitory action of the antisense oligodeoxynucleotide (AS-ODN) to the expression of VEGF protein by radiotherapy in the prostate cancer cell line (PC3M).

METHODSTo observe the changes of serum VEGF in the prostate cancer patients during radiotherapy dynamically and the inhibitory action of the antisense oligodeoxynucleotide to the expression of VEGF by radiotherapy in PC3M.

RESULTSThe changes of serum VEGF in three patients receiving radiotherapy had been observed continuously. The levels of serum VEGF began to increase when the patients received radiotherapy and rised up to peak value after fifteen days, then declined to the range of pre-radiotherapy. Irradiating the PC3M cells with X-rays significantly increased the VEGF expression and secretion. The expression of VEGF protein in the group treated by VEGF AS-ODNs and X-ray irradiation decreased significantly than the group treated only by X-ray irradiation.

CONCLUSIONSThe induction of VEGF protein expression by X-ray irradiation in tumor cells may result in the increasing of the VEGF in the prostate cancer patients during radiotherapy and the induction can be blocked by VEGF AS-ODNs.

DNA, Antisense ; pharmacology ; Endothelial Growth Factors ; antagonists & inhibitors ; blood ; genetics ; Gene Expression ; drug effects ; radiation effects ; Humans ; Intercellular Signaling Peptides and Proteins ; blood ; genetics ; Lymphokines ; antagonists & inhibitors ; blood ; genetics ; Male ; Prostatic Neoplasms ; blood ; pathology ; Radiotherapy ; adverse effects ; Tumor Cells, Cultured ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factors

In order to develop potent antidiabetic agents that have inhibitory effect to a-glucosidase, twelve β-acetamido ketone derivatives such as N-{[(substituted-4-oxo-thiochroman-3-yl)phenyl]-methyl}acetamide are designed and synthesized through one-pot Dakin-West reaction. Their chemical structures are confirmed by 1H NMR, 13C NMR, IR and HR-MS. In vitro α-glucosidase inhibition assays of compounds 4a-41 were carried out using glucose oxidase method. The result indicated that most of them possess inhibitory activity in vitro. Compound 4k showed the most potent inhibitory activity with 87.3% inhibition of α-glucosidase at the concentration of 5.39 mmol x L(-1). The structure-activity relationship of these β-acetamido ketone derivatives was discussed preliminarily. Moreover, the molecular docking method was used to study the interaction mode of compound 4k and α-glucosidase. Our results will be helpful for designing of α-glucosidase inhibitors in the future.
Acetamides ; Glycoside Hydrolase Inhibitors ; chemical synthesis ; pharmacology ; Hypoglycemic Agents ; chemical synthesis ; pharmacology ; Molecular Docking Simulation ; Structure-Activity Relationship ; alpha-Glucosidases ; metabolism

Objective To investigate the effectiveness of digital technology in repairing wounds of the lower leg and foot with perforator pedicled propeller flaps.Methods Eighteen patients with wounds of the lower leg and foot were returned to the hospital for review and evaluated retrospectively.The wounds were repaired by using the perforator pedicled propeller flaps from January,2013 to February,2014.There were 11 males and 7 females,with an average age of 27 years (range,6-48 years).Including 6 cases of injuries caused by the spokes and the Achilles heel of soft tissue defects.Five cases of infection after internal fixation of calcaneal fractures induced skin necrosis,2 cases of dorsal skin defects caused by heavy injured,5 cases of foot and ankle soft tissue defects caused by car accidents.All wounds were associated with exposure of tendon.The wounds area were ranged from 2.5 cm×5.0 cmn to 4.0 cm× 15.0 cm.The course of disease was from 3 hour to 35 days.Computed tomography angiography (CTA) was performed preoperatively,the appropriate perforator was selected and the CTA data were imported into the Mimics 15.0 software for the location of the perforator vessel and the design of the propeller flap,and simulate flap cut and transfer.The flap was obtained according to preoperative plan during operation.The flap size ranged from 4.0 cm×7.0 cm to 5.0 cm ×20.5 cm.These flaps included terminal branch of the peroneal artery perforator in 14 cases,posterior tibia artery perforator in 4 cases.All patients were followed up at regular intervals.Results The reconstruction of Mimics 15.0 software could confirm the perforator vessels origin,vascular distribution,diameter,and the cutting length.The rotation direction of the flap could be simulated preoperatively,which was consistent with the actual observation intraoperative.The donor sites were sutured directly.One case suffered from vascular crisis in 1 day was cured by the removal of part of the suture,massage and bloodletting.All cases were followed-up for 1 month to 16 months,and all flaps survived well and pediele were smooth with a satisfied appearance.The patient were extremely satisfied with the results for repair.Conclusion The preoperative individualization design of the perforator pedicled propeller flaps can be realized through CTA combined with Mimics 15.0 software.It can reduce the risk of operation.

BACKGROUND: Studies have shown that dental pulp stem cells have high proliferation and multi-directional differentiation abilities and can differentiate into a variety of cells under certain conditions. At present, the use of dental pulp stem cells to construct tissue-engineered dentin-pulp complex is expected to become a new strategy for human dental defect repair . OBJECTIVE: To observe the effect of dental pulp stem cells on the repair of rat tooth defects by construction of tissue-engineered dentin-pulp complex. METHODS: Twenty-four Sprague-Dawley rats were used to make animal models with dental pulp removal, and then model rats were randomly divided into model group and transplantation group. Rats in the transplantation group were subjected to tissue-engineered dentin-pulp complex transplantation, and those in the model group given no treatment. Tooth samples were collected at 3, 5, 7 weeks post transplantation and observed using hematoxylin-eosin staining and immunofluorescence staining. The dentin thickness of rats was measured by Image Pro Plus 6.0 image software system. RESULTS AND CONCLUSION: (1) Dental pulp cells was mostly spindle/oval-shaped and partially polygonal. The third generation of cells with long spindle shape showed fibrous growth and uniform morphology. Findings from immunohistochemical staining showed spindle-shaped deep-colored cells with oval nuclei stained as dark blue were identified as fibroblast-like cells, and were positire for vimtin. (2) Findings from hematoxylin-eosin staining showed vacuolar degeneration of the cells, and hbdestroyed pulp tissue and debris, irregular cord-like tissue, and a large amount of red blood cells and inflammatory cells in the pulp cavity, accompanied by clearly visible vascular dilation. Seven weeks after transplantation, a bundle of odontoblasts were visible in the matrix-like tissues of the dentin, and there was a distinct boundary between the original dentin and regenerated dentin. (3) Findings from immunofluorescent staining showed that after dentin-pulp complex transplantation, the number of cells in the pulp cavity increased significantly at 3 weeks, and there was also a substantial increase in dental pulp cells at 5 weeks that were distributed on the wall of the pulp cavity. Compared with the model group, the dentin thickness in the transplantation group was significantly higher at each time after transplantation (P < 0.05), and in the transplantation group, there was also a significant difference in the dentin thickness at different time points (P < 0.05). To conclude, the tissue-engineered dentin-pulp complex can promote dentin regeneration and repair.