In the field of research and development of biomedicine and drugs, the technologies about the extraction, isolation and purification of proteins play an important role. In recent years, the research on proteins has achieved significant progress, and new understanding of various properties of proteins has been achieved. At the same time, the development of physical, chemical and engineering technologies has also promoted the continuous improvement and promotion of extraction and isolation methodology. This paper reviews the technologies of extraction, isolation and purification according to the sources and classification of protein products.

ObjectiveTo investigate the effects of astragalin on pulmonary edema after acute cervical spinal cord injury in rats.MethodsA total of 200 adult Wistar rats weighing 240-250 g were randomly divided into five groups:astragalin group,low concentration astragalin group,physiological saline group,blank group and sham group,with 40 rats in each group.The rats with cervical spinal cord injury were induced at C7 by modified Allen' s method,with the dropping weight of 10 × 2.5 g · cm.In the sham group,the laminas were removed only,leaving spinal cord at the C7 intact.Each group was further divided into four time points:24 hours,3 days,1 week and 2 weeks after the modeling,with 10 rats in each time point,according to the specimen collection time.Rats were sacrificed at different time points to observe the pathological change of the lung tissue using optical microscope,measure the lung wet/dry weight ratio (W/D) and protein concentrations of the serum and bronchoalveolar lavage fluid and calculate the lung W/D and lung permeability index (LPI).ResultsAt the same instant,the W/D and LPI in the astragalin group and low concentration astragalin group were lower than those in physiological saline group and blank group,with the lowest value in the astragalin group at day 3 after injury ( P ＜0.05 ).ConclusionsRats with acute cervical spinal cord injury may cause pulmonary edema,which can be efficiently alleviated through early use of astragalin.

OBJECTIVETo explore the effect of Qingyi Granule (QYG) on high mobility group box-1 (HMGB1) expressions in liver and renal tissues of severe acute pancreatitis (SAP) rats.

METHODSFifty-four Sprague-Dawley (SD) rats were divided into the sham-operation (SO) group, the SAP group, and the QYG group according to random digits table. Rats in the SAP group were induced by injecting 5% sodium taurocholate (STC). Liver and renal pathological changes were observed by HE staining. Serum contents of amylase (AMS), MDA, IL-1, and HMGB1 were detected by ELISA. HMGB1 protein expressions in liver and renal tissues were tested by immunohistochemistry. HMGB1 mRNA expressions in liver and renal tissues were detected by reversed transcription PCR.

RESULTSThe pathological scores, serum levels of AMS, MDA, IL-1 and HMGB1, and protein and mRNA HMGB1 expressions in liver and renal tissues were increased more obviously in the SAP group than in the SO group (P < 0.05, P < 0.01). All of them could be down-regulated by QYG intervention, with the most significant effect seen at 72 h (P < 0.05, P < 0.01) in a time-effect relationship.

CONCLUSIONSHMGB1 participated in SAP complicated liver and renal injuries. QYG could effectively inhibit HMGB1 expressions, thereby attenuating SAP complicated liver and renal injuries.

Amylases ; Animals ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; HMGB1 Protein ; metabolism ; Interleukin-1 ; Kidney ; metabolism ; Liver ; metabolism ; Pancreatitis ; drug therapy ; metabolism ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Taurocholic Acid

Animals ; Animals, Newborn ; Hippocampus ; cytology ; drug effects ; physiology ; Interleukin-6 ; pharmacology ; Ion Channels ; drug effects ; physiology ; N-Methylaspartate ; physiology ; Neurons ; drug effects ; physiology ; Patch-Clamp Techniques ; Rats ; Rats, Wistar

Objective To observe the distribution of blood vessels in tibial metaphysis in ovariectomized and control mice by micro-CT(?CT),and investigate the relationship between bone regional blood supply and osteoporosis. MethodsForty mice were randomly divided into ovariectomy group(n=20) and control group(n=20).Four weeks after operation,?CT analysis was conducted to observe the bone blood vessel distribution after silicone rubber perfusion,and bone mineral density measurement,?CT bone microarchitecture analysis and biomechanical test were performed. Results Bone mineral density,bone microarchitecture in ?CT analysis,biomechanical properties and bone blood vessel distribution in ?CT analysis of ovariectomy group were significantly lower than those of control group(P

AIM:To study the effect of different surgeries for vision and postoperative complications in leprosy patients with lagophthalmos.METHODS:A retrospective analysis.Totally 68 leprosy patients (97 eyes) with lagophthalmos were selected who treated with different surgeries during May 2007 to September 2015 in our sanatorium.The surgeries included nylon thread correction surgery, tarsorrhaphy, temporalis transposition surgery.We observed and compared the best corrected visual acuity(BCVA) and the rate of postoperative complications, such as epiphora, lacrimal duct injury, hard to open eyes, infection at the inner and lateral canthus and scar formation of lacrimal apparatus.RESULTS:The effect on BCVA of patients treated with nylon thread correction surgery and tarsorrhaphy:the preoperative and postoperative BCVA was significantly different of the two groups (Z=-4.193,-4.213;P<0.05);the preoperative and postoperative BCVA was not significantly different of the patients underwent temporalis transposition surgery (Z=-1.532, P >0.05).The postoperative complications:(1) epiphora:there was 91% in patients underwent nylon thread correction surgery, 89% in patients underwent tarsorrhaphy, 59% in patients underwent temporalis transposition surgery, the difference was statistically significant (χ2=12.198, P<0.05);(2) lacrimal duct injury:there was 66% in patients underwent nylon thread correction surgery, 95% in patients underwent tarsorrhaphy, 41% in patients underwent temporalis transposition surgery, the difference was statistically significant (χ2=22.415, P<0.05);(3) hard to open eyes:there was 56% in patients underwent nylon thread correction surgery, 55% in patients underwent tarsorrhaphy, 22% in patients underwent temporalis transposition surgery, the difference was statistically significant (χ2=8.795, P<0.05);(4) infections:there was 84% in patients underwent nylon thread correction surgery, 3% in patients underwent tarsorrhaphy, 11% in patients underwent temporalis transposition surgery, the difference was statistically significant (χ2=60.858, P<0.05).CONCLUSION:There are certain clinical efficacy for lagophthalmos caused by leprosy after surgery, especially patients after temporalis transposition surgery can close eyes by oneself.And their vision does not change significantly, which suggests that this surgery may prevent exposure injury for cornea.But the iatrogenic injury, the vision decrease in patients underwent other surgeries and the ocular tissue damage, such as epiphora, lacrimal duct injury, hard to open eyes, infection at the inner and lateral canthus and scar formation of lacrimal apparatus should be pay more attention to.

10.3969/j.issn.1007-3969.2013.04.00X

The L-cysteine desulfhydrase gene (cd) of Pseudomonas sp.TS1138 was amplified by PCR,and the amplified gene was recombined in the cloning vector pBluescript SKII.The 1.2kb DNA fragment containing cd was sequenced,and its homology with other desulfhydrases was blast; then the cd was cloned into the expression vector pET-21a(+), and afterward expressed by IPTG inducement.The expression protein was purified by Ni-NTA His-Bind Resin.Then the expression protein was identified by the method of activity staining of desulfhydrase, and the characterization of L-cysteine desulfhydrase and the critical role it played in the L-cysteine biosynthetic pathway were discussed.

The L-ATC hydrolase and L-SCC amidohydrolase which convert L-ATC to L-cysteine in Pseudomonas sp.TS-1138 are purified about 83.9 and 90.3 fold by salting-out method, Sephadex G-75 gel chromatography, DEAE-cellulose 52 ion exchange and Sephadex G-100 gel chromatography, etc. The purified enzyemes are both demonstrated by SDS-PAGE to be a homogeneous protein. Their molecular weight are about 37.5kD and42.8kDa respectively. The optimum reaction temperature are both 35℃, and the optimum pH are 7.0 and 8.0 respectively. The Km of the two enzymes are 0.67 mmol/L and 0.15 mmol/L, and the Vmax are 0.39?10 -3mmol/L?min and 0.42?10 -3mmol/L?min respectively.

Objective To establish a three- dimensional hindlimb gait data toolkit (THGT) for healthy and spinal cord injured (SCI) non-human primate (rhesus monkey) based on Matlab to realize upload of original data, automatic gait division, calculation and drawing of multiple gait parameters, etc. Methods Vicon system was used to collect three-dimensional hindlimb gait data of healthy and SCI (after 6 weeks) rhesus monkey to obtain the kinematics data of both hindlimbs in continuous strides. It was analyzed with THGT to process the gait division, calculation and drawing of multiple gait parameters. Results THGT read the data, distinguished cycles of gait, calculated 140 kinds of gait parameters and drew graphs of the results. Conclusion THGT extends the universality of the Vicon data, realizes automatically gait division and friendly interactive interface, and puts out the visible results.