0.05 for all).Conclusions The allelic frequencies of LPL gene at Pvu Ⅱ locus in Hei Yi Zhuang were different from those in Han,but the genotypie frequencies in Hei Yi Zhuang were not different from those in Han.There was no significant correlation between the polymorphism of LPL gene at Pvu Ⅱ site and the serum lipid levels in two ethnic groups.

OBJECTIVETo investigate the possibility to fabricate a blood vessel scaffold with a combined polymer for tissue engineering.

METHODSA blood vessel scaffold was designed with a combined polymer composed of rabbit vascular smooth muscle cells(VSMCs), collagen and a non-spinning fabric mesh of polyglycolic acid (PGA). VSMCs were implanted into collagen gel and their growth was observed. The mixed solution of VSMCs and collagen was dropped into the tubular scaffold, followed by 7-day culturing.

RESULTSVSMCs formed many prominences after culturing in gelatinous collagen for 3-4 hours. With cells extending, some cells became shuttle- or spindle-shaped. After VSMCs-collagen complex was implanted into the PGA mesh, most of VSMCs remained in the pore of PGA mesh with the formation of gelation. VSMCs could adhere to and grow on the PGA fiber.

CONCLUSIONThe non-spinning PGA porous biodegradable material coated with collagen is a good carrier for VSMCs to adhere and grow.

Animals ; Blood Vessels ; growth & development ; Collagen ; Muscle, Smooth, Vascular ; growth & development ; Polyglycolic Acid ; Polymers ; Rabbits ; Time Factors ; Tissue Engineering ; instrumentation ; methods ; Tissue Scaffolds

Saliva and blood were collected from two patients who had not received post exposure prophylaxis in the cities of Wenzhou and Xinning respectively. Both patients were confirmed as positive for rabies by detection of rabies virus specific nucleoprotein antibodies in the sera by Western Blot. However, rabies virus specific RNA was only identified in the saliva collected from the patient in Wenzhou. Furthermore, the isolate Zhejiang Wz0 (H) was obtained by inoculating one-day-old suckling mice. Both nucleoprotein (N) and glycoprotein (G) genes from the isolate were amplified by RT-PCR and sequenced. Phylogenetic analysis indicated that the isolate belonged to classic rabies virus, and shared a higher homology with the street viruses from dogs in the main endemic areas in China and the street virus from dogs in Indonesia than with other known strains. Further comparison of the deduced amino acid sequences between the isolate and the vaccine strains used in China showed that the virus had a higher level of homology with the vaccine strain CTN than with the other vaccine strains (3aG, PV, PM and ERA). In particular, amino acid residues substitutions located in antigenic site Ⅲ in the G protein, which could react with the neutralizing antibodies, were observed. These results suggested that the virus belonged to the classic rabies virus, and both N and G genes diverged from the current vaccine strains used in China at either the nucleotide or the amino acid level.

OBJECTIVE: To assess the safety and efficacy of gastrointestinal anastomosis with nickel titanium shape memory alloy compression anastomosis clip. METHODS: We randomized 51 patients to undergo gastrointestinal anastomosis with stapler (n=25) and nickel titanium compression anastomosis clip (n=26) respectively. The following parameters were recorded to evaluate the safety and efficacy: mean hospitalization time, anastomotic complication, first post-operation flatus and bowel movement, and extrusion of the clip. RESULTS: Anastomotic complications such as leakage, stenosis and obstruction were not observed in both groups. There were no significant differences in the first post-operation flatus time and bowel movement time between the 2 groups (P>0.05). The clip was expelled with stool within 9-15 d. CONCLUSION Compression anastomosis clip is safe and effective.
Anastomosis, Surgical ; instrumentation ; methods ; Anastomotic Leak ; prevention & control ; Digestive System Surgical Procedures ; methods ; Female ; Gastroenterostomy ; instrumentation ; methods ; Gastrointestinal Diseases ; pathology ; surgery ; Humans ; Male ; Nickel ; Surgical Staplers ; Titanium

Objective To evaluate the incidence of postoperative hemorrahage in children undergoing adenoidectomy , and to discuss its possible causes. Methods Included in this study were children who underwent adenoid and/or tonsil surgery at Shenzhen Chilidren's Hospital between January 2004 and November 2009. The change of hemoglobin ( Hb) and hematocrit ( Hct) were retrospectively analysed. The blood loss was estimated by the change of Hct. Results There were 2078 cases that accomplished the inclusion criteria in the period of study. Ten children bled 0. 5 - 4. 0 hours after surgery,without superfluous hemorrahage during the operation and post-tonsillectomy. This represented an incidence of 0.48% of immediate postoperative haemorrhage among the 2078 procedures analyzed. Statistical differences were found between boys (0.21%) and girls (1.10%, x2 =5.597, P<0.05). The change of Hb and Hct was positively correlated (r = 0. 95 , P < 0. 01) , the blood loss was positively correlated with the bleeding time ( r = 0. 66, P < 0. 05 ) . The causes of postoperative hemorrhage were coagulation system deficits, chronic nasopharyngitis, deficient hemostasis and immoderate ravage. To control the postoperative hemorrhage, 2 postnasal packing under topical anaesthesia and 8 electrocautery under general anaesthesia were applied. Conclusions Poor operative technique and deficient hemostasis are the major causes of primary hemorrahage. Prompt operation to control the postoperative bleeding should be done 2 hours after bleeding under general anesthesia in order to avoid severe complications.

OBJECTIVETo construct the recombinant adenovirus containing herpes simplex virus-1 virion protein (VP) 22 and human microdystrophin gene, then the adenovirus was transfected into C2C12 myoblast and studied on the property of protein transduction with VP22-mediated microdystrophin in C2C12 myoblast.

METHODSThe full-length VP22 cDNA was obtained from recombinant plasmid pSINrep5-VP22 with PCR, and the product was directionally inserted into pShuttle-CMV to acquire the plasmid pCMV-VP22. Microdystrophin cDNA was obtained from recombinant plasmid pBSK-micro digested with restrictive endonuclease NotI, and the product was directionally inserted into pCMV-VP22 to acquire the plasmid pCMV-VP22-MICDYS. The plasmid of pCMV-VP22-MICDYS was lined with Pme I, and the fragment containing VP22-microdystrophin was reclaimed and transfected into E1 coli BJ5183 with plasmid pAdeasy-1. After having been screened by selected media, the extracted plasmid of positive bacteria was transfected into HEK293 cells with liposome and was identified by observing the cytopathic effect of cells and by PCR method to acquire the recombinant adenovirus Ad-VP22-MICDYS. Finally, the C2C12 myoblast were transfected with the recombinant adenovirus Ad-VP22-MICDYS and Ad-MICDYS, and the expression of microdystrophin was detected by RT-PCR, Western blot and immunocytochemistry.

RESULTSThe recombinant adenovirus including VP22 and microdystrophin gene was successfully constructed. VP22 transferred VP22-microdystrophin fused protein from infected C2C12 myoblast into uninfected cells and enhance the expression of microdystrophin in myoblast.

CONCLUSIONSRecombinant adenovirus containing VP22 and microdystrophin gene was constructed successfully. VP22 can enhance the expression with microdystrophin in myoblast. It lays the foundation for further studying on VP22-mediated recombinant including microdystrophin gene to cure Duchenne muscular dystrophy.

Adenoviridae ; genetics ; physiology ; Animals ; Cell Line ; Dystrophin ; genetics ; metabolism ; Genetic Vectors ; genetics ; metabolism ; Humans ; Mice ; Myoblasts ; metabolism ; virology ; Simplexvirus ; genetics ; metabolism ; Transduction, Genetic ; Viral Structural Proteins ; genetics ; metabolism ; Virion ; genetics ; metabolism

OBJECTIVETo study the relationship between the polymorphism of the variable number of tandem repeats region 3' of the apolipoprotein B gene (3'APOB-VNTR) and serum lipid levels in the Guangxi Heiyi Zhuang population.

METHODSA total of 548 people of Heiyi Zhuang nationatity were surveyed by a cluster sampling. Epidemiological data were collected and serum lipid and apolipoprotein levels were measured. The genotypes and alleles of the 3' APOB-VNTR were determined by polymerase chain reaction combined with gel electrophoresis, and then analyzed by direct sequencing in the most common alleles. The results were compared with those in 496 people of Han nationality also live in that district.

RESULTSThere were 19 alleles of the 3'APOB-VNTR in both ethnic groups. They were hypervariable elements (HVEs) 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62 and 64, but HVEs 56 and 58 in Heiyi Zhuang nationality and HVEs 48 and 62 in Han nationality were not be detected. The most common allele is HVE32 in Heiyi and Zhuang nationality (25.9%), and HVE34 in Han nationality (27.2%). The frequencies of HVEs 26, 30, 46, heterozygote, and short alleles (< 38 repeats, S) were higher in Heiyi Zhuang nationality than in Han nationality, whereas the frequencies of HVEs 34, 38, 40, homozygote, and long alleles (>or= 38 repeats, L) were lower in Heiyi Zhuang nationality than in Han nationality. The levels of total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C) and apolipoprotein (apo) B in Heiyi Zhuang nationality were higher in VNTR-LS (carrier of one long and one short alleles) than in VNTR-LL genotypes (the individual carrying two long alleles) genotypes. The levels of TC, triglycerides, HDL-C and apo B in Heiyi Zhuang nationality were also higher in homozygotes than in heterozygotes. There were no significant differences in the detected lipid parameters between the VNTR-SS (carrier of two short alleles) and VNTR-LS or VNTR-LL genotypes in both ethnic groups.

CONCLUSIONThe 3'APOB-VNTR polymorphism is found to be significant difference between Heiyi Zhuang nationality and Han populations, and is associated with the serum lipid levels in Heiyi Zhuang nalionality but not in Han nationality.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Apolipoproteins B ; genetics ; China ; ethnology ; DNA ; analysis ; Ethnic Groups ; genetics ; Female ; Gene Frequency ; Humans ; Lipids ; blood ; Male ; Middle Aged ; Minisatellite Repeats ; genetics ; Polymorphism, Genetic ; Tandem Repeat Sequences ; Young Adult

AIMAnnonaceous acetogenin 89-2 was obtained from atemoya plant. To investigate the effect of 89-2 on experimental chemotherapy against xenografts derived from multidrug resistant KBv200 cells and parental drug-sensitive KB cells.

METHODSCytotoxicity was determined by tetrazolium (MTT) assay. The models of KB and KBv200 xenografts in nude mice were established to investigate the effect of 89-2 on experimental chemotherapy against cancer in vivo. Mechanistic experiments were conducted to examine the function of P-gp by Fura 2-AM assay.

RESULTSThe compound 89-2 showed potent cytotoxicity in KBv200 and KB cells, and the mean IC50 of 89-2 to KBv200 and KB cells was 48.7 and 64.6 nmol.L-1, respectively. The IC50 of 89-2 to multidrug resistant (MDR) cells was similar to that to the parental drug-sensitive cells (P < 0.05). In the models of KBv200 and KB cell xenografts in nude mice, 89-2 (0.90 mg.kg-1, q2d x 6) exhibited 52.3% and 56.5% in inhibiting the growth of xenografts, respectively. The toxicity was endurable. The intracellular accumulation of Fura-2 in KBv200 cells increased to 1.66, 2.03, and 2.74-fold, respectively, by addition of 12.8, 64 and 320 nmol.L-1 of 89-2.

CONCLUSIONBoth MDR KBv200 cells and parental drug-sensitive KB cells were sensitive to the treatment of 89-2 in vitro and in vivo. The mechanism of overcoming MDR was associated with the decrease of P-gp function MDR cells.

4-Butyrolactone ; analogs & derivatives ; isolation & purification ; pharmacology ; ATP-Binding Cassette, Sub-Family B, Member 1 ; metabolism ; Animals ; Annona ; chemistry ; Antineoplastic Agents, Phytogenic ; isolation & purification ; therapeutic use ; Cell Division ; drug effects ; Disease Models, Animal ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; Drugs, Chinese Herbal ; isolation & purification ; therapeutic use ; Fatty Alcohols ; isolation & purification ; pharmacology ; Humans ; KB Cells ; Male ; Mice ; Mice, Nude ; Neoplasm Transplantation ; Neoplasms, Experimental ; drug therapy ; Plants, Medicinal ; chemistry ; Xenograft Model Antitumor Assays

OBJECTIVETo develop a rapid genetic diagnosis technique for the patients with hereditary hearing loss by screening hot spots of mutations, namely 235delC of the GJB2 gene, IVS7-2A>G of the SLC26A4 gene, and 1555A>G of mitochondrial 12S rRNA.

METHODSMultiple PCR amplification of the three fragments covering the expected mutations in GJB2, SLC26A4 and 12S were carried out and the amplified products were analyzed by restriction fragment length polymorphism (RFLP).

RESULTSEighteen homozygous and 18 heterozygous 235delC, 2 homozygous and 13 heterozygous IVS7-2A>G, and 8 homogeneous 1555A>G were detected in the 200 patients with hearing loss. All the results were confirmed by sequencing. The detection rate of the three mutant alleles was 21.7% (71/400 + 8/200 = 0.217) and the genetic diagnosis rate was 14% [(18+2+8)/200 = 0.14].

CONCLUSIONIt is a convenient, efficient and economical method to screen the hot spots of mutation in the patient with hereditary hearing loss by using PCR-RFLP.

Adolescent ; Adult ; Asian Continental Ancestry Group ; genetics ; Child ; Child, Preschool ; Connexin 26 ; Connexins ; genetics ; Female ; Hearing Loss ; genetics ; Humans ; Male ; Membrane Transport Proteins ; genetics ; Mutation ; Polymerase Chain Reaction ; methods ; Polymorphism, Restriction Fragment Length ; Young Adult

Objective To construct a recombinant plasmid expressing human microdystrophin gene fused with VP22 ofhmnan herpes simplex virus 1(HSV-1),and investigate the expression of microdystrophin in vitro and the characteristics of VP22-mediated protein transduction.Methods Full length HSV-1 VP22 gene was amplified by PCR from the plasmid pSINrep5-VP22 and cloned into the eukatyotic expression vector pAVXl to conslruct recombinant plasmid pAVP22.Microdystrophin cDNA was obtained from the recombinant plasmid pBSK-MICRO digested with Not Ⅰ,and the product Was inserted into plasmid pAVP22 to construct the plasmid pAVP22-MICDYS. 3T3 cells were transfected with pAVP22-MICDYS,and the expression of microdystrophin was detected by RT-PCR,Western blotting and immtmocytochemislry.The supematant of 3T3 cells transfected with pAVP22-MICDYS were collected to infect human mesenchymal cells(MSCs),and the expression of the fusion protein VP22-microdystrophin in the cells was detected using immunohistochemistry to assess VP22-mediated protein transduction. Results The recombinant plasmid expressing microdystrophin-VP22 fusion gene capable of in vitro expression of the fusion protein was constructed successfully.VP22 was shown to enhance the expression efficiency of microdystrophin in 3T3 cells and transduce VP22-microdystrophin fusion protein from 3T3 cells to human MSCs. Conclusion The recombinant plasmid expressing microdystrophin-VP22 fusion gene with protein transduction capacity provides an important basis for further study of the fusion protein in treatment of Duchenne muscular dystrophy.