ObjectiveTo optimize human Hexastatin gene,to express,purify protein and conduct activity experimental research,and to provide a theoretical basis for further study of Hexastatin.MethodsHuman Hexastatin gene was optimized and synthesized.It was connected to the pET28a expression vector,induced to express by isopropyl β-D-1-thiogalactopyranoside(IPTG),and optimized induction conditions.After the ultrasonication of bacterial cells and inclusion bodies,the recombinant fusion protein was purified with Ni-NTA chromatographic column,analyzed and identified by SDS-PAGE and Western Blot,and conduct activity experimental research in vitro by MTT.ResultsConstructed production was pET28a-Hexastatin expression plasmid.The human Hexastatin protein was expressed in E.coli BL21 the high level and accounted for 45.1％ of the total bacterial protein.The purification of recombinant protein purified with Ni-NTA chromatographic column reached 90％,and the concentration was 80 μg/ml.Human Hexastatin protein can restrain the growth of C6,MCF-7 and human vascular endothelial cell (HMEC) cells,and inhibition ratio reach to 72.9％±3.6％,48.8％±2.9％,52.7％±2.5％,respectively through MTT test.ConclusionThe optimized human Hexastatin protein was expressed successfully,which confirmed the inhibition to tumour cells.It is a new way for anti-angiogenesis therapy of tumour.

OBJECTIVETo study the morphological and anatomical characteristics of Coptis deltoidea based on the research of its breeding system.

METHODThe flowering process of C. deltoidea was observed dynamically and the out-crossing index was estimated, using Motic BA200 microscope to observe the morphologic characteristics of the mixed bud, stolon and pollen was observed microscopically, and compared with those of C. chinensis.

RESULTThe results showed that the breeding system of C. deltoidea was facultative hybridization, it could bloom normally and fruit while cannot form seeds. And there were no statistical differences in the number of stamens, pistils, ovules, pollens in a stamen, pollens in a flower, pollen-ovule ratio, and the pollen grain size between C. deltoidea and C. chinensis. However the pollen of C. deltoidea developed anomaly, most of the pollen grains in it were surface depression, the texture was unsharp or broken, the pollen could not germinate. The lateral bud on the lower side of the mixed bud formed in the bud stage. Pericyclic fibers in the stolon (the vegetative propagation branch) shaped like a cap, and all the shaped-caps nearly formed a ring.

CONCLUSIONThe abnormal development of the pollen could be the mainly cause to the no formation of seeds in C. deltoidea. The lateral bud forming in the bud stage then developing the stolon is the characteristics of the asexual propagation. Pericycle fibers in the stolon nearly forming a ring is a secondary character to accommodate the vegetative propagation of C. deltoidea.

Breeding ; Coptis ; anatomy & histology ; genetics ; growth & development ; Flowers ; anatomy & histology ; genetics ; growth & development ; Pollen ; anatomy & histology ; genetics ; growth & development

The aim of the present study was to observe whether protein tyrosine kinase (PTK) within the nucleus tractus solitarius (NTS) was involved in the regulation of ventilatory responses of peripheral chemoreflex. The experiments were performed on anesthetized, immobilized and artificially ventilated rabbits. Peripheral chemoreflex was elicited by ventilating the animal with 10% O2-balance 90% N2. Changes in the peak amplitude and frequency of integrated phrenic nerve activity were observed. The ventilatory responses of peripheral chemoreflex following 0.1 microl microinjection within the NTS of either PTK inhibitor genistein (10 mol/L), AMPA glutamate receptor inhibitor CNQX (10 mol/L),or inactive PTK inhibitor daidzein (10 mol/L) were recorded. The results are as follows: Both genistein and CNQX attenuated the ventilatory responses of peripheral chemoreflex, while no changes occurred following daidzein. The amplitude of integrated phrenic nerve discharge and the phrenic burst frequency were decreased by (-21.77+/-6.93)% and (-24.70+/-7.61)% respectively after administration of genistein. CNQX resulted in similar decreases in the amplitude of phrenic nerve discharge (-27.13+/-7.63)% and the burst frequency (-21.34+/-4.88)%. In addition, the inhibitory effects of CNQX and genistein were the same whether they were applied alone or one after another, indicating that they had no cooperative effects. The results obtained suggest that PTK within the NTS regulates the peripheral chemoreflex control of respiration and that this regulation of PTK may be mediated through the phosphorylation of AMPA receptors in NTS neurons.
Animals ; Brain Stem ; enzymology ; physiology ; Chemoreceptor Cells ; physiology ; Female ; Male ; Protein-Tyrosine Kinases ; physiology ; Rabbits ; Receptors, AMPA ; physiology ; Respiration ; Solitary Nucleus ; enzymology

OBJECTIVETo screen the representing components in different germplasms of Canarium album and determine the content by HPLC for laying the foundation for the quality specification.

METHODGallic acid and scopoletin and scoparone were chosen as the main chemical components for clearing heat and relieving sore throat by reviewing literature and qualitative experiment. Gallic acid was determined according the method of Galla chinensis in Chinese pharmacopoeia. Scopoletin and scoparone were determined by HPLC on C18 column (4.6 mm x 250 mm, 5 microm) and the mobile phase was acetonitrile-water-acetic acid (16:84:0.5) with the detection wavelength at 345 nm and column temperature at 35 degrees C , the flow rate was 1.0 mL x min(-1).

RESULTThe resolution is good for gallic acid and scopoletin and scoparone. The standard curve in detection range showed good linear relation, the average recoveries were 100.22%, 101.22% and 100.42%, respectively, RSD were all less than 2.0%. The content of gallic acid in the large fruit C. album from Guangdong was higher than that in the small fruit C. album, while the result was contrary in samples from Sichuan. The content of total coumarins in the small fruit C. album was generally higher than that in the large fruit C. album, which from Sichuan was higher than those from Guangdong.

CONCLUSIONThe efficacies of clearing heat and relieving sore throat in C. album were formed by synergistic effect of gallic acid and scopoletin and scoparone. The small fruit C. album was better than the large fruit C. album as medicinal materials, the results provide reliable basis for quality control in medicinal materials and its preparation.

Burseraceae ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Coumarins ; therapeutic use ; Drug Synergism ; Drugs, Chinese Herbal ; chemistry ; Fever ; drug therapy ; Gallic Acid ; therapeutic use ; Hot Temperature ; Humans ; Pharyngitis ; drug therapy ; Scopoletin ; therapeutic use

To study the anti-coagulant effect and influence of danggui Sini decoction (DSD) on rat's plasma endogenous metabolites by animal experiment and ¹H-NMR based metabolomics method. After intragastric administration of Danggui Sini Decoction for 7 days, Plasma thrombin time (TT) was measured. Rat plasma metabolic fingerprint in two groups was analyzed using ¹H-NMR, based on which the principal component analysis( PCA) and orthogonal partial least-squares discriminant analysis(OPLS-DA) models for metabonomic analysis. Potential biomarkers were screened by using variable importance in the projection (VIP) and T test. DSD could prolong TT of the rat significantly (P < 0.05). Compared with control group, six kinds of endogenous metabolites in DSD group change significantly (P < 0.05), among which isobutyrate, carnitine and phenylalanine content had an upward trend (P < 0.01) and lysine, Histidine and cholesterol content had a downward trend (P < 0.05). It is likely that carnitine, phenylalanine, Histidine and cholesterol are the potential metabolic markers in the anti-coagulant process and DSD affects the platelet aggregation and the expression of tissue factor and fiber protease by regulating the energy, amino acid and lipid metabolism.
Animals ; Anticoagulants ; chemistry ; Blood Coagulation ; drug effects ; Drugs, Chinese Herbal ; chemistry ; Female ; Male ; Metabolomics ; methods ; Proton Magnetic Resonance Spectroscopy ; methods ; Rats

Objective To study the effects of dexamethasone (DEX) and/or hyperbaric oxygen (HBO) on the subdermal vascular network (SVN).Methods SVNs were selected on dorsal skin flaps of 40 Wistar rats.The animals were divided randomly into a control group,a DEX group,a HBO group and a HBO+DEX group.Cranially based,2.5 cm?10 cm dorsal SVN skin flaps were sharply incised and elevated between the dartos and SVN,then sutured to their beds.On the 7th postoperative day,the surviving flap area was measured along with the number of new capillary,the thickness of meat tissue and the number of infiltrated neutrophilie granulocytes in the SVN skin flap.Results The mean surviving flap area for the control group was 7.90 cm~2,for the DEX group it was 10.48 cm~2,for the HBO group 15.53 cm~2,and for the DEX+HBO group it was 15.58 cm~2.The improvement in surviving flap area was highly statistically signifieant compared with the control.The improvement was also statistical- ly significant when the HBO group or HBO+DEX group was compared with the DEX group.However,no statistically significant difference was found between the HBO group and HBO+DEX group.Conclusion In a rat dorsal skin flap model,DEX or HBO improved skin flap viability,but DEX alone is not as efficacious as HBO or as DEX+ HBO.DEX plus HBO showed no additive beneficial effect over HBO alone.

VEGF nanoparticle (VEGF-NP) was prepared by a multi-emulsification technique using a biodegradable poly-dl-lactic-co-glycolic (PLGA) as matrix material. The nanoparticles were characterized for size, VEGF loading capacity, and in vitro release. VEGF-NP and naked VEGF plasmid were intramuscularly injected into the ischemia site of the rabbit chronic hindlimb ischemia model and the efficiency of VEGF-NP as gene delivery carrier for gene therapy in animal model was evaluated. Gene therapuetic effect was assessed evaluated by RT-PCR, immunohistochemistry and angiography assay. The average size of VEGF-NP was around 300 nm. The encapsulation efficiency of VEGF was above 96%. Loading amount of VEGF in the nanoparticles was about 4%. In vitro, nanoparticles maintained sustained-release of VEGF for two weeks. Two weeks post gene injection the capillary density in VEGF-NP group (81.22 per mm2) was significantly higher than that in control group (29.54 mm2). RT-PCR results showed greatly higher VEGF expression in VEGF-NP group (31.79au * mm) than that in naked VEGF group (9.15 au * mm). As a carrier system for gene therapy in animal model, VEGF-NP is much better than naked DNA plasmid. The results demonstrate great possibility of using NP carrier in human gene therapy.
Animals ; Disease Models, Animal ; Gene Transfer Techniques ; Genetic Therapy ; Genetic Vectors ; chemistry ; Lactic Acid ; chemistry ; Nanoparticles ; chemistry ; Plasmids ; Polyglycolic Acid ; chemistry ; Rabbits ; Vascular Endothelial Growth Factor A ; genetics

Objective To make the cost-effectiveness analysis of pembrolizumab and platinum chemotherapy as the first-line treatment for advanced non-small cell lung cancer(NSCLC)in the population with tumor proportion score(TPS)≥1%of PD-L1,and provide some reference for the clinical use and future price negotiation of pembrolizumab.Methods Based on Pubmed database,the published RCT literatures of pembrolizumab were analyzed,and the survival data were extracted,combined with the treatment plan of a tertiary hospital,the Markov model were established to simulate the cost and health effectiveness of patients for twenty years,and the stability of the model was tested by one-way sensitivity analysis and probability sensitivity analysis.Results Twenty years later,the cost-effectiveness ratio of pembrolizumab group and chemotherapy group was ￥58 517.60/quality adjusted life month(QALM)and ￥41 213.08/QALM.Compared with the chemotherapy group,the incremental cost effective ratio(ICER)was ￥104 485.36/QALM.Conclusion When the willingness to pay(WTP)value was ￥30 902/QALM,the pembrolizumab therapy was not more cost-effective advantages than platinum chemotherapy,and the sensitivity analysis showed that the results of the model were relatively stable.

OBJECTIVETo analyse the related epidemic factors of rabies and to provide basic evidence for developing rabies control and prevention programs in China.

METHODSPossible factors that causing rabies epidemics in the southern part of China were obtained through collecting both epidemic and surveillance data of rabies in 2005, and to analyse every factor by software of Excel 2002.

RESULTSThe number of rabies cases was 2548 in 2005. Among 885 cases being investigated, 60.56%, 49.04%, 96.16% had not had any chance to receive treatment, vaccine or immunoglobulin. Among the exposed persons, 89.95% received treatment on wounds and 94.93% were immunized, and 8.56% received immunoglobulin. 94.86% of them accomplished the full rabies post-exposure prophylaxis course, but 8.56% of them did not accomplish it. 88.50% of the cases and 92% of the persons under exposure were bitten by dogs. The density of dogs was 3.20-13.37 per 100 persons. The vaccination coverage rates among dogs were 5.31% -75.11% with a positive rate of 2.93 % -6.40%.

CONCLUSIONFactors as: low rate of post-exposure prophylaxis and failed to have finished the necessary post-exposure treatment (PET) of the cases, nonstandard PET in hospital or local clinic, low rabies vaccination coverage and high infectivity of rabies virus in animal host (mainly for dog) might be responsible for rabies epidemics in China, in 2005.

Animals ; Bites and Stings ; China ; epidemiology ; Disease Outbreaks ; Dogs ; Humans ; Population Surveillance ; Post-Exposure Prophylaxis ; Rabies ; epidemiology ; Rabies Vaccines ; administration & dosage

BACKGROUNDSmall noncoding microRNAs regulate gene expression in cardiac development and disease and have been implicated in the aging process and in the regulation of extracellular matrix proteins. However, their role in age-related cardiac remodeling and atrial fibrillation (AF) was not well understood. The present study was designed to decipher molecular mechanisms underlying age-related atrial structural remodeling and AF.

METHODSThree groups of dogs were studied: adult and aged dogs in sinus rhythm and with persistent AF induced by rapid atrial pacing. The expressions of microRNAs were measured by quantitative real-time polymerase chain reaction. Pathohistological and ultrastructural changes were tested by light and electron microscopy. Apoptosis index of myocytes was detected by TUNEL.

RESULTSSamples of atrial tissue showed the abnormal pathohistological and ultrastructural changes, the accelerated fibrosis, and apoptosis with aging and/or in AF dogs. Compared to the adult group, the expressions of microRNAs-21 and -29 were significantly increased, whereas the expressions of microRNAs-1 and -133 showed obvious downregulation tendency in the aged group. Compared to the aged group, the expressions of microRNAs-1, -21, and -29 was significantly increased in the old group in AF; contrastingly, the expressions of microRNA-133 showed obvious downregulation tendency.

CONCLUSIONThese multiple aberrantly expressed microRNAs may be responsible for modulating the transition from adaptation to pathological atrial remodeling with aging and/or in AF.

Age Factors ; Animals ; Apoptosis ; Atrial Fibrillation ; etiology ; Atrial Remodeling ; Connective Tissue Growth Factor ; physiology ; Dogs ; Electrocardiography ; Fibrosis ; In Situ Nick-End Labeling ; MicroRNAs ; analysis ; physiology ; Myocardium ; pathology ; ultrastructure