Objective To prepare a redox-responsive doxorubicin-loaded nanoparticle,and to study its in vitro realease behavior and targeting effect on heptoma cells.Methods Cystamine was grafted on the side chains of hyaluronic acid with 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride/N-hydroxysuccinimide catalyst,and then β-cyclodextrin (β-CD) was conjugated on the amine groups of the cystamine by Schiff's base reaction to prepare β-CD modified hyaluronic acid (HACD).The HACD/DOX nanoparticles were prepared by encapsulating DOX into HACD using dialysis method.The drug loading,encapsulation efficiency,particle size and distribution,zeta potential and other physical and chemical properties,as well as in vitro drug release behavior of the HACD/DOX nanoparticles were characterized.The cytotoxicity of HACD/DOX nanoparticles to HepG2 cells was studied by cell counting kit-8 (CCK-8) method.The targeting effect of HACD/DOX nanoparticles on HepG2 cells was studied using flow cytometry and confocal laser scanning microscopy (CLSM).Results HACD were successfully synthesized,which could carry DOX to form uniform homogeneous nanoparticles.The drug loading of DOX in the nanoparticles was (16.1±0.2)％ and the encapsulation efficiency was (64.2±0.9)％.The transmission electron microscope images indicated that the shape of the HACD/DOX nanoparticles was homogeneous sphere.The results of granularity analysis showed that the average size of the HACD/DOX nanoparticles was (203.1 ±2.5) nm with a narrow size distribution (PDI =0.202).The zeta potential of the HACD/DOX nanoparticles was (-29.1±0.8) mV.The in vitro release behavior of the nanoparticles exhibited obvious redox-sensitivity.The results of in vitro cytotoxicity showed that the blank carrier material HACD had no obvious toxicity to hepatoma cells,and the HACD/DOX nanoparticles could effectively kill hepatoma cells with the 0.38 μg/ml half maximal inhibitory concentration (IC50) value at 48 h.Flow cytometry and CLSM results demonstrated that the HACD/DOX nanoparticles could target hepatoma cells through the mediating effect of hyaluronic acid.Conclusions The prepared HACD/DOX nanoparticles have suitable particle size,high drug loading and encapsulation efficiency,and can release DOX under the stimulation of reducing agent.These nanoparticles have obvious targeting effect on hepatoma cells,which is expected to be applied as the drug delivery system of hepatocellular carcinoma (HCC) therapy.

ObjectiveTo investigate the effect of operation treating spinal multi-level tuberculosis.Methods45 patients with multi-level spine tuberculosis were treated with debridement completely and anterior or lateral-anterior intervertebrae autograft. Of them, 5 patients added to anterior instrumentation.Results45 patients had been followed up for 12 to 40 months.The back pain of 40 cases were relieved within 5 weeks after operation, erythrocyte sedimentation rate decreased 3 weeks after operation. 38 patients who involved in kyphosis decreased their kyphosis mostly and only one patient lost 3 degrees in follow-up.15 patients who involved in neurological deficits improved one or two grades (Frankel). Grafts fused in 44 patients and there were no recurrent in follow-up.Conclusions Operative treatment is efficacious to multi-level body spine tuberculosis.

Objective To explore clinical significence of soluble vascular cell adhesion molecular(SVCAM - 1),immunoglobulin A (IgA) anti- endothelial cells antibodies(AECA) in Henoch-Schonlein purpura(HSP) and their relationships. Methods SVCAM-1 were detected by ELISA between 55 cases with HSP (40 cases in acute stage and 15 cases in recovery) and 20 controls. Results The levels of serum SVCAM - 1 of the HSPN cases were significantly increased compared with the other HSP. In addition, the serum level of SVCAM - 1 in acute stage HSP children was significantly higher than that in recovery stage and normal controls. IgA AECA positive rate in HSPN was higher than that of without nephritis group. The level of serum SVCAM - 1 in IgA AECA positive children was higher than that of IgA AECA negative children. Conclusion SVCAM - 1 and IgA AECA may participate in the pathogegesis of HSP, and the level of molecule is correlated with the progress of HSP.

Objective To study the clinical features,diagnosis and treatment of eosinophilic granuloma of bone(EGB).Methods Twelve cases(8 male and 4 female)of EGB and their medical record,imaging examinations,follow-up data were reviewed,and the lesion sites,symptom and clinical features were analyzed with reference to relative literature.Results Nine unifocal cases were found in the cervical spine(3 cases),the thoracic spine(1 case),the lumber spine(1 case),the femur(1 case),the metacarpal bones(1 case),and the ilium(2 cases).Three multifocal cases were found in the thoracic and lumber spine(1 case),in the femur,the multiple ribs,the skull,the scapula(1 case)and in the skull,the femur,the ilium(1 case).The most common presenting symptom was pain at the lesion site and restricted motion was obviously in the cervical lesion.The case of the thoracic lesions was found neurologic deficit.Osteolytic destruction was found in the radiologic examination in EGB and the periosteal reaction was found in the long bone lesions,and vertebra plana was considered as typical cha-racteristics in the spine lesion.The lesions not in the spine were diagnosed by biopsy,exclusive methods and close following-up were performed in the spine lesions.Two unifocal were healed spontaneously and three were performed with curettage with one bone grafting.Multifocal and the lesions in the spine underwent chemotherapy.All cases were followed up and no recurrence was found.Conclusions EGB is commonly solitary and not seldom in the spine.Topical pain may be chief complaint.Osteolytic image was found in the radiologic examination.Biopsy and exclusive methods combining closely follow-up were used for diagnosis.EGB is self-limited observation,so curettage or chemotherapy can be used in treatment,and prognosis is good.

Dorsal root ganglion (DRG) neurons are primary sensory neurons that conduct neuronal impulses related to pain, touch and temperature senses. To comprehensively identify proteins of plasma membrane (PM) from small amount of dorsal root ganglion (DRG) neurons, a proteomics strategy that utilizes aqueous polymer two-phase partition in combination with differential velocity centrifugation was adopted to enrich the PM, followed by SDS-PAGE, CapLC-MS/MS and bioinformatics analysis. Western blot analysis showed that the concentration of PM in purified plasma membrane(PPM) was 2.3 times higher than that in crude plasma membrane(CPM), 15 times higher than that in whole tissue lysate (WTL). By searching against the rat IPI protein sequence database, a total of 729 non-redundant proteins were identified from the PM preparation, of which 547 had a gene ontology (GO) annotation indicating a cellular component, and 159 (21.8%) were unambiguously identified as PM proteins. A data set of plasma membrane proteins of DRG as well as a tool to study PM proteins were provided in a small amounts of sample.

Objective To understand the pathogen distribution situation among the children inpatients with hand ,foot and mouth diseases(HFMD) in Guiyang area during 2012 to provide the basis for the diagnosis ,treatment and prevention .Methods The data in 3 179 cases of HFMD were collected .The fluorescence quantitative RT‐PCR was adopted to perform the genotyping on universal enterovirus ,enterovirus 71(EV71)and Coxsackie virus A16(CA16) .Results A total 3 179 samples of HFMD were col‐lected ,among them ,151 cases (4 .75% ) were CA16 positive ,331 cases (10 .41% ) were EV71 positive ,7 cases (0 .22% ) were CA16 and EV71 co‐infection ,and 897 cases(28 .22% )were the other enterovirus .The whole year had 2 peaks of onset ,which were April to July and October to November .The onset age focused on the children aged under 5 years old (96 .16% ) ,among them ,0-3 years old had the highest onset ,moreover male children were more than female .Conclusion The etiology distribution of children HFMD in Guiyang area during 201 was dominated by the other genotypes of enterovirus and EV71 .

Objective To explore clinical significance of immunoglobulin G(IgG) subclass and the relationship with urine enzyme series and four microalbumen in Henoch - Schonlein purpura nephritis(HSPN). Methods IgG subclass, urine enzyme series, microal-bumen were detected between 28 cases with HSPN and 20 controls. Results The levels of IgG1, IgG2 in HSPN significantly decreased compared with the normal controls. In addition, the levels of IgG, were negative related to microalbumin(MA) and alkaline phosphalase (ALP) in HSPN, P

AIM:To study the feasibility of recombinant adeno-associated virus(rAAV)as a vector to transfer the green fluorescent protein(GFP)gene as a target gene into rabbit retina.METHODS:Intravitreal injection of rAAV-gfp was performed in either eye for each rabbit with the other eye taken as control.At the 3rd,7th,and 14th day after injection,the eyeballs were removed,and the retinas were flat-mounted on glass slides to inspect the retinal fluorescence,respectively.RESULTS:After intravitreal injection of rAAV-gfp,the presence of fluorescent spots in the cytoplasm of retinal cells indicated that GFP gene was efficiently transferred and expressed in the rabbit retina.CONCLUSION:Recombinant adeno-associated virus is a reliable and simple vector for transferring target gene,e.g.,GFP gene,to the retina.

Objective To study the cytotoxicity against brain microvessel endothelial cells and blood compatibility in rats of OX26 conjugated endomorphin (EM) loaded hyperbranched polyglycerols-poly(lactic-co-glycolic acid)(HBPG-PLGA) nanoparticles.Methods Prepared nanoparticles were divided into group B (HBPG-PLGA nanoparticles),group EP (EM-HBPG-PLGA nanoparticles) and group OEP (OX26-EM-HBPG-PLGA nanoparticles).The cytotoxicity against brain microvessel endothelial cells (BMECs) of nanoparticles of different groups were measured by MTT test,haemolysis test,normal haemotological parameter and several primary items of coagulation system were tested after nanoparticles of different groups and different dosages injection on rats.Results ①All the three groups of nanoparticles induced decreased cell viability in a dose dependent manner in MTT test,whereas all groups of nanoparticles showed low cytotoxicity against the BMECs during 30 to 600 μg/ml.②There was no significant difference in haemolysis ratio (P＞0.05) and normal haemotological parameter (P＞0.05).③There was no significant difference between the low dosage of all the three groups of nanoparticles and the control group on the function of coagulation system in rats (P＞0.05).④Compared with C group,high dose groups demonstrated longer prothrombin time (PT),activeated partial thromboplasting time (APTT) and lower fibrinogen (Fbg) (P＜0.05).At the same time,compared with the low dose subgroups,PT and APTT were prolonged,Fbg significantly decreased in the high dose subgroups (P＜0.05 or P＜0.01).Conclusion OX26 coupled with EM-HBPG-PLGA nanoparticles showed low cytotoxicity against BMECs and had no significant effect on the coagulation system in rats with low concentration and low dosage.

Objective To study the distribution of genotypes of hepatitis C virus in Guizhou and its relationship between infec-tious route of genotype and age ,gender was analyzed .Methods Serum specimens in this study were obtained from 198 patients , whose anti-HCV and HCV RNA were positive .A reverse transcriptase PCR(RT nested-PCR)assay using conserved primers de-duced from the core-envelope 1(C-E1)region of the hepatitis C virus(HCV)genome was employed to amplify a 474-nucleotide-long fragment .Phylogenetic analysis of the C-E1 sequences was conducted by direct sequencing of the RT-PCR products and alignment with published HCV subtypes in GenBank .Subtypes of the samples were determined by nucleotide sequencing followed by composi-tion of a phylogenetic tree .Results Among the 198 patients surveyed ,genotype 1a was detected in 4 cases(2 .0% ) ,genotype 1b in 71 cases(35 .9% ) ,genotype 2a in 9 cases(4 .6% ) ,genotype 3a in 29 cases(14 .7% ) ,genotype 3b in 47 cases(23 .7% ) ,genotype 6 a in 37 cases(18 .7% )and genotype 6d in 1 cases(0 .5% ) .Genotype distribution on gender had no statistical significance(P>0 .05) , and its distribution on people with different ages had statistical significance(P<0 .05) ,and its distribution on patients with different infectious routes was significantly different(P<0 .05) .Conclusion The major genotypes of HCV are 1b ,3b ,6a and 3a in Guizhou , and genotype 1a is predominant .Genotypes 1a ,2a and 6d exist too .Genotypes of patients infected with HCV are related to their in-fectious routes ,and the HCV genotypes are in a great variety .