The fermentation conditions which affect C-15? hydroxylation o f 18-methyl-estr- 4-ene-3,17-dione were investigated. As the key step in the Hydroxylation, the dissolution of substrate was focused on. Tween80, MeOH, DMSO,?-CD and 2-HP?CD were studied to improve the dissolvability of 18-met hyl-estr-4-ene-3,17-dione.The other factors such as pH, substrate concent ration and aeration strategies which affected conversion rate were also resea rched. As a result, the conversion rate can be up to 60% in shake flask and ach i(eve 50% in fermentor,which would overcome the disadvantage of 15?-hydroxyl -18-methyl-estr-4-ene-3,17-dione chemosynthesis and provide a good techn ics to industry.

The synthesized full-length hGLP-1 gene was cloned into pET-32a(+) to get the recombinant plasmid pET32-GLP-1, which could express a fusion protein containing thioredox, hexahistidine, and rhGLP-1 consecutively. The recombinant plasmid containing hGLP-1 was transformed into E.coli BL21 (DE3) and expressed by IPTG induction. The fusion protein was purified from lysates with Ni?IDA His?Bind affinity chromatography. rhGLP-1 with the purity of 90% was achieved after enterokinase digestion, Ni?IDA His?Bind affinity chromatography again, then was concentrated by ultrafiltration. The purified rhGLP-1 showed a single band on IEF gel with an isoelectric point between pH5.2 and pH5.85. ESI mass spectrometry showed that the molecular weight was 3355.0kDa as expected. rhGLP-1 was digested with trypsin followed by mass analysis and the peptide mapping produced two expected fragments with the molecular weights of 2097.7kDa and 1005.5kDa, respectively. The purified rhGLP-1 also showed obvious biological activity for both lowering plasma glucose and stimulating insulin secretion in mice.

The objective of this work was to investigate the effect of six individual strains of fungi on the reduction of gossypol levels and nutritional value during solid substrate fermentation of cottonseed meal (CSM). Six groups of disinfected CSM substrate were incubated for 48 h after inoculation with either of the fungi C. capsuligena ZD-1, C. tropicalis ZD-3, S. cerevisae ZD-5, A. terricola ZD-6, A. oryzae ZD-7, or A. niger ZD-8. One not inoculated group (substrate) was used as a control. Levels of initial and final free gossypol (FG), crude protein (CP), amino acids (AA) and in vitro digestibility were assayed. The experiment was done in triplicate. The experimental results indicated that microbial fermentation could greatly decrease (P<0.05) FG levels in CSM. The detoxification efficiency differed between the species of microorganisms applied. From the perspective of reducing CSM potential toxicity, C. tropicalis ZD-3 was most successful followed by S. cerevisae ZD-5 and A. niger ZD8. They could reduce FG levels of CSM to 29.8, 63.07 and 81.50 mg/kg based on DM (dry matter), respectively, and their detoxification rate were 94.57%, 88.51% and 85.16%, respectively. If crude protein, amino acids content and their in vitro digestibility were also taken into account, A. niger ZD-8 may be the best choice. The CP content of CSM substrate fermented by C. tropicalis ZD-3 and A. niger ZD-8 were improved by 10.76% and 22.24%; the TAA (total amino acids) contents were increased by 7.06% and 11.46%, and the EAA (essential amino acids) were raised by 7.77% and 12.64%, respectively. Especially, the levels of methionine, lysine and threonine were improved greatly (P<0.05). The in vitro CP digestibility of CSM fermented by C. tropicalis ZD-3 and A. niger ZD-8 was improved by 13.42% and 18.22%, the TAA were increased by 17.75% and 22.88%, and the EAA by 16.61% and 21.01%, respectively. In addition, the in vitro digestibility of methionine, lysine and threonine was also improved greatly (P<0.05).
Amino Acids ; analysis ; Cottonseed Oil ; chemistry ; metabolism ; Digestion ; Fermentation ; Fungi ; metabolism ; Gossypol ; analysis ; Nutritive Value ; Plant Proteins ; analysis

OBJECTIVEThe aim of the present study was to prepare uniform-sized silybin loaded poly (lactic-co-glycolic acid) (PLGA) microspheres in study of silybin with stainless steel membrane.

METHODSilybin PLGA microspheres were prepared by stainless steel membrane emulsification. The preparation conditions were optimized by single-factor test and orthogonal experiment, and evaluating the mean diameters, the particle size distribution, drug loading, entrapment efficiency and morphology of microsphere.

RESULTPrepared microspheres were round and surface was smooth. The mean diameter was (4.961 +/- 0.56) microm. The span was (1.75 +/- 0.18). The entrapment efficiency was (54.997 +/- 4.05)% and the average drug loading was (23.6 +/- 1.70)%.

CONCLUSIONThe stainless steel membrane emulsification can be used to prepare the silybin PLGA microspheres. The mean diameters of the silybin PLGA microspheres can be controlled in certain level. Stainless steel membrane emulsification has great potentiality exploitation and utilization.

Drug Compounding ; methods ; Emulsions ; chemistry ; Lactic Acid ; chemistry ; Microspheres ; Particle Size ; Polyglycolic Acid ; chemistry ; Silymarin ; chemistry ; Stainless Steel ; chemistry

OBJECTIVETo explore the operative skills and effect of unilateral pedicle screw combined with contralateral percutaneous transfacet screws fixation in treating degenerative low lumbar disease.

METHODSFrom January 2009 to December 2011,22 patients with degenerative low lumbar disease were treated with transforaminal lumbar interbody fusion, during the operations, unilateral pedicle screw and contralateral percutaneous transfacet screw fixation were performed. There were 16 males and 6 females, aged from 32 to 71 years old with an average of (51.1 ± 10.6) years, including single segment in 20 cases and two segments in 2 cases. Clinical effects were evaluated according to visual analogue score (VAS) and Oswestry Disability Index (ODI).

RESULTSAll patients were followed up from 1 to 2.5 years with an average of 18 months. One case complicated with leakage of cerebrospinal fluid after operation and 1 case with lower limb pain of decompression-side on the 3rd day after operation. Twenty-two patients got bony fusion. There were no instability and evidence of instrument failure during follow-up. The VAS and ODI score decreased from preoperative 8.24 ± 0.72, 36.72 ± 6.84 respectively to 3.18 ± 0.66, 4.36 ± 1.12 at the final follow-up (P < 0.05).

CONCLUSIONUnilateral pedicle screw combined with contralateral percutaneous transfacet screw fixation is safe and feasible surgical technique in treating low lumbar degenerative disease. It has advantages of little trauma, rigid fixation, high fusion rate, and less complication. etc.

Adult ; Aged ; Biomechanical Phenomena ; Female ; Humans ; Intervertebral Disc Degeneration ; physiopathology ; surgery ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Pedicle Screws ; Spinal Fusion ; methods

OBJECTIVETo observe the clinical value of protoparaxotril saporlirs (PTS) combined with aspirin in the secondary prevention of cerebral infarction.

METHODSThe 140 patients with cerebral infarction were collected, among them the 120 patients during recovery stage were equally assigned to three groups by randomized, single blinded and open controlled principle, and they were treated respectively by PTS (A), aspirin (B), and PTS plus aspirin (C) for 6 months. The other 20, who couldn't or were unwilling to use aspirin, were arranged in group D for control. The platelet aggregation rate, incidence of stroke recurrence, gastrointestinal adverse reaction and the NIHSS scores of patients were observed during the six-month period of treatment.

RESULTSAs compared with group D, the lowering amplitude of platelet aggregation rate after treatment in the three treatment groups were significantly higher (P < 0.01). Comparison of platelet aggregation rate between group A and B showed significant difference after 3-month treatment (P < 0.05), but the difference became insignificant after 6-month treatment (P > 0.05). The incidence of stroke recurrence in the group A, B and C was 18.9%, 13.2% and 10.8% respectively, which showed no significant difference among them, but all were significantly lower than that in the group D (44.4%, P < 0.05). NIHSS scores in group A and C were significantly lower than in group B (P < 0.01); and the occurrence of gastrointestinal reaction was significantly lower in group A (P < 0.01).

CONCLUSIONLong-term application of PTS has the effects for preventing stroke recurrence, lowering gastrointestinal adverse reaction and improving patients' neural function in patients with stroke. As used in combination with aspirin, it shows potential practical importance in the clinical secondary prevention of stroke.

Aged ; Aged, 80 and over ; Aspirin ; administration & dosage ; adverse effects ; Cerebral Infarction ; drug therapy ; physiopathology ; prevention & control ; Drug Therapy, Combination ; Female ; Humans ; Male ; Middle Aged ; Platelet Aggregation ; drug effects ; Sapogenins ; administration & dosage ; adverse effects ; Secondary Prevention

Prawn white spot syndrome is caused by the pathogen prawn white spot syndrome virus (WSSV). VP19 is a vesicle membrane protein of WSSV. HyNPV (Hybrid of AcNPV and BmNPV) constructed by the recombination of BmNPV and AcNPV is a new hybrid virus having both of their advantages. The recombinant transfer vector pBlueBicHisC-vp19 and recombinant baculovirus HyNPV-VP19 were constructed on the basis of the successful cloning of VP19. Newly-molted silkworms Bombyx mori of fifth instar were inoculated by the recombinant virus. SDS-PAGE and Western blotting analysis showed a specific band, about 21kD, which was consistent with the expectation suggesting that the WSSV-VP19 gene was successfully expressed in silkworm bodies.
Animals ; Baculoviridae ; genetics ; metabolism ; Bombyx ; genetics ; metabolism ; virology ; Genetic Vectors ; Penaeidae ; virology ; Recombinant Fusion Proteins ; genetics ; Viral Envelope Proteins ; genetics ; metabolism ; Virus Replication ; White spot syndrome virus 1 ; genetics

OBJECTIVETo study the incidence of injuries among residents living in the communities in Zhejiang.

METHODSA household questionnaire survey was conducted to 16 899 residents in four communities.

RESULTSThe incidence of injuries was 16.11% in total, higher in urban area than in rural area and higher in men than in women. The leading causes of injuries were falls (4.80%), knife-cutting or by sharp articles (3.86%), traffic accident (2.07%), collision (1.99%), scalds (1.13%) and animal bites (1.06%). Peak incidence of age group was different in different injuries, such as traffic accident was high in 45 - 60 age group and falls was high in 5 and 60 age group. Direct economic loss for each case with injury would cost 324.21 RMB yuan.

CONCLUSIONInjury was common and frequently occurred among residents at the community level which seriously on danger their health and families.

Accidental Falls ; statistics & numerical data ; Accidents, Traffic ; statistics & numerical data ; Adolescent ; Adult ; Aged ; Animals ; Bites and Stings ; epidemiology ; Burns ; epidemiology ; Child ; China ; epidemiology ; Community Health Services ; Dogs ; Female ; Humans ; Male ; Middle Aged ; Prevalence ; Wounds and Injuries ; epidemiology ; mortality

OBJECTIVETo investigate the biomechanical differences between the surgery and adjacent segments of intervertebral discs in the lower lumbar spine, which were implanted with Coflex into the segments of L4, and L5S1, respectively.

METHODSThree finite-element models (the model of the intact lower lumbar sacrum,the L4.5 and L5S1 segments implanted by Coflex) were developed, respectively. According to the spinal three-column loading theory, three models were forced by the physiological loads of upright standing, flexion and extension. The stress of the different areas of the disc annulus, the changes of intervertebral dorsal height and the degree of nucleus pulposus pressure were compared and analyzed.

RESULTSCoflex implanted into the L4.5 and L5S1 segments in compression and extension could both decrease the stress of the posterior area of intervertebral disc in the surgery segment, resist the changes of the intervertebral disc dorsal height and reduce the perssure of nucleus pulposus. Furthermore, the stress of the L5S1 segment decreased when Coflex fixed the L4.5 segment in extension. However, when Coflex fixed the L5S1 segment, the stress of L4.5 segment had no significant changes.

CONCLUSIONCoflex fixing the L4,5 and L5S1 segments can effectively decrease the stress of the surgery segmental discs, respectively. Furthermore, Coflex fixing L4,5 segment may play a biomechanical role in reducing the stress of L5S1 segment.

Adult ; Biomechanical Phenomena ; Finite Element Analysis ; Humans ; Internal Fixators ; Intervertebral Disc ; physiopathology ; surgery ; Lumbar Vertebrae ; surgery ; Male ; Stress, Mechanical

This 6-week study was conducted to evaluate the effects of seven different levels of dietary chromium (Cr) (0, 75, 150, 300, 450, 600, and 1 200 ppb Cr) in the form of Cr nanoparticle (CrNano) on growth, body composition, serum hormones and tissue Cr in Sprague-Dawley (SD) rats. Seventy male SD rats (average initial body weight of (83.2+/-4.4) g) were randomly assigned to seven dietary treatments (n=10). At the end of the trial, body composition was assessed via dual energy X-ray absorptiometry (DEXA). All rats were then sacrificed to collect samples of blood, organs and tissues for determination of serum hormones and tissue Cr contents. The results indicated that lean body mass was significantly increased (P<0.05) due to the addition of 300 and 450 ppb Cr from CrNano. Supplementation of 150, 300, 450, and 600 ppb Cr decreased (P<0.05) percent body fat significantly. Average daily gain was increased (P<0.05) by addition of 75, 150, and 300 ppb Cr and feed efficiency was increased (P<0.05) by supplementation of 75, 300, and 450 ppb Cr. Addition of 300 and 450 ppb Cr decreased (P<0.05) the insulin level in serum greatly. Cr contents in liver and kidney were greatly increased (P<0.05) by the addition of Cr as CrNano in the dosage of from 150 ppb to 1 200 ppb. In addition, Supplementation of 300, 450, and 600 ppb Cr significantly increased (P<0.05) Cr content in the hind leg muscle. These results suggest that supplemental CrNano has beneficial effects on growth performance and body composition, and increases tissue Cr concentration in selected muscles.
Animals ; Body Composition ; drug effects ; Body Weight ; drug effects ; Chromium ; administration & dosage ; chemistry ; pharmacokinetics ; Dose-Response Relationship, Drug ; Hormones ; blood ; Male ; Nanoparticles ; administration & dosage ; ultrastructure ; Organ Specificity ; Particle Size ; Rats ; Rats, Sprague-Dawley ; Tissue Distribution