Objective To study the effect of cold-processing technology on the composition of water-soluble proteins in ground dragon and their activities of thrombolysis and anti-tumor activity.Methods Ground dragon was respectively processed by traditional processing technology and cold-processing technology.The latter was done by the liquid nitrogen rapid freezing technology and lyophilizing technology.By means of qualitative and quantitative tests,SDS-PAGE and the software of ImageTool were applied to analyzing the difference of protein components in ground dragon from the two processing techniques.Fibrin plate method and thrombolysis test in vitro were carried out to determine the difference of the two processing techniques for the fibrinolytic activity.Micro-observation and MTT methods were used to assess the differences of antihepatic cancer activity between the traditional and the cold-processing groups.Results Compared with the traditional group,the components of water-soluble proteins in the ground dragon processed by cold-processing technique were more abundant than traditional one,and the number of protein bands and concentration in SDS-PAGE gel were more and much higher.Although the two kinds of protein solution from the different processing techniques had the visible fibrinolytic activity and antihepatic cancer activity,both activities of cold-processing technique were markedly higher than those in the traditional group.Conclusion Effects of different processing techniques of ground dragon on fibrinolytic activity and anti-tumor activity are significant.On protecting the bioactivity of proteins in ground dragon,the cold-processing techniques is advantage to the traditional processing technology.

Objective To purify a Rhizoma Pinelliae protein,determine its inhibiting activity and mechanism to trypsin,and analyze its N-terminal amino acid sequence. Methods Active Rhizoma Pinelliae protein was purified from the 40% (NH_4)_2SO_4 sediment of the crude extracted protein from Rhizoma Pinelliae by affinity chromatography of trypsin-Sepharose CL 4B and gel filtration of Sephadex G-50;12% of SDS-PAGE was used for determining the purity of the purified Rhizoma Pinelliae protein and estimating its molecular weight;N-terminal amino acid sequence of active Rhizoma Pinelliae protein was analyzed by Edman degradation. Results The purified active Rhizoma Pinelliae protein showed a sin-gle band on SDS-PAGE gel,which estimated molecular weight was about 1. 4?10～4,its specific activity was 1 059. 012 U/mg,and the yield was about 24.40%;The N-terminal sequence of the preceding six amino acid residues of active Rhizoma Pinelliae protein was DPVVDG;The quality inhibiting ratio of active Rhi- zoma Pinelliae protein to trypsin was 1:4. 72,and the Ki value ,the inhibition constant was about 7. 17 ?10～(-6) mol/L. Conclusion Active Rhizoma Pinelliae protein is a kind of serine protease inhibitor,a com-petive inhibitor to trypsin.

Objective To determine the lumbrukinase contents and its hydrolysates in processed earthworm,set up a new evaluating method of dot blotting to evaluate the qualities of those processed Chinese medicines as earthworm. Methods Contents of lumbrukinase and its hydrolysates in processed earthworm were determined by dot blotting with rabbit antibody of anti-lumbrukinase as the first antibody. Blotting signal was analyzed by Quantity One software. Results Contents of lumbrukinase and its hydrolysates in processed earthworm of Pheretima aspergilum,P. pingi,Drawida japonica,and D. gisti were 125.23,118.34,81.05,and 72.13 ?g/g,respectively. Conclusion There was no significant difference between the processed earthworms of P. aspergilum and P. pingi in the content of lumbrukinase and its hydrolysates,both of which were higher than those of the other two species in Shandong. Theoretically,the processed products of P. pingi could be used as the succedaneum of P. aspergilum in the clinical of traditional Chinese medicine. Dot blotting,as a convenient and accurate method,could be broadly used for the evaluation of the processed products of Chinese crude drugs similar to earthworm.

A novel teaching pattern of the experimental course of biotechnology was discussed in the paper.The teaching model of fusing the experimental items with concrete research subjects was proposed and put to practice.The result showed that the new teaching pattern could convert precious intelligence and material resources into valuable scientific results,and efficiently avoid the consumption of experiment materials.Under the experimental teaching pattern,students were activated to initiatively enjoy the interest of scientific research and exploration.The new teaching pattern provided students with good chance for the cultivation of their independent development and creative thinking and comprehensive quality.

The four properties theory of traditional Chinese medicine (TCM) is an important basis for the guidance of TCM clinical medication.However,TCM modernization has produced a large number of active substances with unknown medicine properties.Therefore,it is urgent to build a new drug development system guided with TCM four properties theory.From the view of that four properties of TCM must have their material basis,and based on the known knowledge on TCM four properties,this paper proposed strategies of new drug research by cloning TCM partial components based on theTCM model of standard level property. A certain number of representative standard level property TCM drugs were selected to conduct system separation for the establishment of aTCM model of standard level property with the proportion and content of each separated component as a reference.The standard model was used as a boundary.Contents and proportions of the experimental TCM separation components deviated from thisstandard model were classified as hot or cold properties.From cloning of the partial components of TCM with hot or cold property medicine,the quality standards of partial components of experimental TCM were also studied,in order to investigate their corrective actions and mechanisms.The TCM partial components were expected to form a new TCM drug with the four properties theory of TCM as guidance.

Objective To study the effects of treadmill training on the recovery of neurological function and the expression of IGF-1/IGF-1Rin the brain poat focal cerebral ischemia.Metheods Forty-two male,adult Sprague-Dawley rats were randomly divided into a sham group(n=6),a model group(n=18)and a treadmill exercises group(n=18).Focal cerebral ischemia was induced in the latter two groups by right middle cerebral artery occlusion(MCAO)using a modified Longa's metheod.The rats in the treadmill exercises group were given treadmill training 6 days a week for 4weeks beginning 24 h after the MCAO. Rats in the sham group and model groups were reared in standard cages and could move frddly. Before being sacrificed at the 7th 14th and 28th day after the MCAO,the rats' neurological function were tested giving modified neurological weverity scores (mNSS). The expression of IGF-1/IGF-1R was detected using the reverse transcriptsae polymersae chain reaction and immunohistochemical methods.Results Neurological runction in the exercises group at the 14th and 28th day was better than in the control group,and IGF-1 and IGF-1Rexpression were significcantly upregulated on the 7th,14th and 28th day after the MCAO.Conclusion Treadmill training can improve neurological function by upregulating the expression of IGF-1 and IGF-1R in the ischemic brain, at least in rats.

AIM　To study the activation of choline on M3-R in heart and observe the hemodynamic changes of rat and rabbit. METHODS　A cardiac catheter was inserted into the left ventricular cavity via the right carotid artery, then the HR, LVSP, LVEDP, and ±dp/dt were measured using a polygraph system. RESULTS　Choline was shown to decrease the hemodynamic assessments, such as HR, +dp/dt, LVSP and LVEDP. while the M3-R antagonist 4-DAMP (4-diphenylacetoxy-N-methylpiperidine-methiodide) showed little effect on these assessments. It was found to reverse the hemodynamic effects of choline. CONCLUSION M3 receptor agonist can produce negative inotropic and chronotropic effects on the heart of rat and rabbit.

AIM: To explore the effects of adipose-derived stem cell (ADSC) transplantation on the expression of IL-10 amd TNF-α after cerebral ischaemia in Middle cerebral artery occlusion (MCAO) rats. METHODS: 72 male adult Sprague-Dawley rats were randomly divided into 4 groups: sham group, MCAO group, Vehicle group and ADSC group (n=18). Rat's cerebral ischemia model was established by MCAO with Longa' s method. ADSC were labeled by DAPI before transplantation. One day after MCAO, 30 μL of cell suspension containing 1×10~6 ADSCs were injected into the lateral ventricle of ADSC group and the same dose of PBS was given to the Vehicle group. At day 4, day 7 and day 14 after MCAO, the rats were decapitated to detect the expression of IL-10 and TNF-α in ischaemic rat' s brain by ELISA, immunohistochemistry and RT-PCR. RESULTS: Compared with sham group, the expression of IL-10 and TNF-α significantly up-regulated at 4 d, 7 d of MCAO group(P<0.05). There was no statistical difference of IL-10 and TNF-α expression between MCAO and vehicle group ant any time point(P>0.05). Compared with Vehicle group, the expression of IL-10 significantly up-regulated while TNF-α expression significantly decreased of ADSC-treated group at any timepoint post MCAO(P<0.05). CONCLUSION: The transplantation of ADSC could up-regulate the expression of IL-10 and down-regulate the expression of TNF-α in MCAO rat' s brain, which might contribute to its protective role upon cerebral ischaemia.

Objective· To investigate the prospective value of early postoperative PI-APD in children with hydronephrosis.Methods· Data of children with hydronephrosis who underwent pyeloplasty in Xinhua Hospital,Shanghai Jiao Tong University School of Medicine between Jan 2012 to Nov 2015 was collected.PI-APD was divided into 3 categories (≤ 19％,19％＜PI-APD＜40％ and ≥ 40％).The relationship between PI-APD value and the degree of renal function (DRF) and dilation recovery after surgery was analyzed.Results· There were 360 children with hydronephrosis.The median follow-up was 20 months.The PI-APD value (3 months after pyeloplasty) was positively correlated with the degree of DRF recovery (r=0.631,P=0.000).Five patients received redo-pyeloplasty.PI-APD of all these patients was ＜19％.Conclusion· PI-APD is a new feasible ultrasound parameter in pyeloplasty followup.PI-APD ≥ 40％ at the first post-operative visit predicts pyeloplasty success.PI-APD ≤ 19％ indicates close follow-up after operation.PI-APD can also help select children at high risk for repeat intervention after pyeloplasty.

OBJECTIVETo research on the substantial foundation of the medical speciality of Chinese traditional medicines from immunogenicity.

METHODControl antigen with hot nature was prepared from the mixture of the aqueous extracts of three Chinese traditional medicines with three typical hot nature of Alpinia officinarum, Cinnamomum cassia and Curculigo orchioides, while that with cold nature prepared with Rheum palmatum, Anemarrhena asphodeloides, Coptis chinensis, and polyclonal antibody was prepared by immunizing rabbit with control antigen. Dot blotting was performed between the polyclonal antibody of control antigen and the aqueous extracts of nine Chinese traditional medicines on a piece of PVDF membrane, and the blotting signals were analyzed by the software of Quantity One.

RESULTBlotting signals with hot control antigen of nine Chinese traditional medicines in descending were Zingiber officinale, Aconitum carmichaeli, Eucommia ulmoides, Fraxinus rhynchophylla, Lonicera japonica, Anemarrhena asphodeloides, Coptis chinensis, Rheum palmatum and Phellodendron chinense, which degree of similarity to control antigen in peak value were 57.33%, 43.56 %, 34.16%, 30.2%, 28.81%, 26.53%, 21.68%, 17.62% and 14.85%, respectively. Blotting signals with cold control antigen were Rheum palmatum, Anemarrhena asphodeloides, Coptis chinensis, Phellodendron chinense, Zingiber officinale, Lonicera japonica, Fraxinus rhynchophylla, Eucommia ulmoides and Aconitum carmichaeli in descending, of which degree of similarity to cold control antigen in peak value were 55.22%, 54.23%, 46.72%, 34.08%, 30.3%, 24.48%, 24.33%, 20.35% and 15.17%, respectively. Results of cluster analysis with Wistar's method showed that nine medicines were classified into two groups, one group included Phellodendron chinense, Anemarrhena asphodeloides, Coptis chinensis, Rheum palmatum, another was Zingiber officinale, Aconitum carmichaeli, Eucommia ulmoides, Fraxinus rhynchophylla, Lonicera japonica.

CONCLUSIONBlotting signals of nine medicines with control antigen regularly varied with the alteration of medicine nature. The more similarity degree of the tested medicine to control antigen was smaller, the more distance of the tested medicine to control antigen was further. Dot immunoblotting was a practical and effective new method in researching the substantial foundation of the medical speciality of Chinese traditional medicines.

Animals ; Antibodies, Monoclonal ; immunology ; Antibody Formation ; Antigens ; immunology ; Blotting, Western ; Cold Temperature ; Drugs, Chinese Herbal ; analysis ; chemistry ; classification ; Hot Temperature ; Immunoblotting ; methods ; Male ; Medicine, Chinese Traditional ; methods ; Rabbits