Objective To comprehensively grasp the grassroots medical institutions blood transfusion(blood) management present situation,put forward improvement strategy for the universality of existence weak link,the stand-ardized management of clinical blood transfusion to strengthen grass -roots medical institutions to provide necessary theoretical support.Methods 62 grassroots medical institutions blood transfusion(blood)in Yantai district were selected as the research subjects,to set the planning,layout,basic construction standards,related technology and equipment application aspects of quantitative evaluation,preliminary conclusions.Results By examination of 62 grassroots medical institutions transfusion quality management level both in terms of hardware and software facilities, emergency plan formulation,system file support informatization construction,staff education training,etc,found many problems,there was a certain gap between the existing department of blood transfusion standards.Conclusion The current grassroots medical institutions transfusion management is conditioned by the objective conditions,there are many problems,such as not improved,will cause a certain hidden trouble on the safety of clinical blood transfusion. Therefore,should combine their own reality,strengthen the quality of blood transfusion management system construc-tion,and give full play to the supervision and management of the hospital transfusion quality management committee. At the same time,the local administrative department of health also should formulate corresponding policies,ensure transfusion management level significantly increased.

Objective To construct recombinant adenovirus vector carrying vascular endothelial growth factor 165(Ad-VEGF165),amplify the adenovirus vector in 293T cells,transfect Ad-VEGF165 into the bone marrow stromal cells(bMSCs) of Wistar rats,and then to assay the expression of VEGF165.Methods VEGF165 obtained by PCR was digested and inserted into adenovirus shuttle plasmid pAdTrack-CMV to generate recombinant plasmid pAdTrack-VEGF165,and then the Pme I-linearized plasmid pAdTrack-VEGF165 was electroporated into E.coli BJ5183 cells that had been electroporated adenovirus backbone plasmid pAdEasy-1.The identified recombinant plasmid pAdEasy-VEGF165 DNA was digested with Pac I and transfected into 293T cells to package adenovirus,followed by identification of the recombinant adenovirus by means of observation of the enhanced green fluorescence protein(EGFP) expression under fluorescent microscope.After amplified in 293T cells,the obtained adenovirus were transfected into 293T cells again,and EGFP expression was detected.Ad-VEGF165 transfected bMSCs were cultured in vitro.The expression of VEGF165 in bMSCs after transfection was determined by observing the expression of EGFP and detected by RT-PCR.ELISA method was applied to assay the secretion of VEGF165.Results Recombinant adenoviral VEGF165 was constructed successfully,which was confirmed by restriction enzyme digestion,gene sequencing and EGFP expression.EGFP expression could be observed under fluorescent microscope,and the expression of VEGF165 was confirmed by RT-PCR.ELISA analysis showed the quantity of expression of VEGF165 in transfection group was higher than that in control group(P

Objective To detect the gene mutation of thyroid hormone receptor β ( TRβ ) in a family with thyroid hormone resistance syndrome.Methods The genomic DNA was extracted from peripheral blood leukocytes of the patient and his 5 family members.The exons 1-10 ofTRβ gene were amplified by PCR.The products of PCR were sequenced directly to detect the gene mutation.Results Two members of this family were confirmed to have the C y A transition mutation at nucleotide 1642 site within exon 10 of TRβ gene,which was a missense mutation causing the substitution of Proline to Threonine (P453T).The mutation was Heterozygous.Conclusions It was confirmed that the patient has TRβ gene mutation P453T in exon 10.The mutation may lead to the occurrence of thyroid hormone resistance syndrome.

Objective To investigate the association between polymorphisms of thyroid-stimulating hormone receptor(TSHR)gene intron 1(rs179247, rs12101261)and Graves′ disease(GD)in the China Han population from Xuzhou city, Jiangsu Province. Methods Total 1 066 GD patients and 1 107 control subjects were recruited for genotyping by Taqman probe technique on Fluidigm EP1 platform. Meanwhile, serum concentrations of thyroid hormone and TSH receptor antibodies(TRAb)were determined. Results The rs179247_A, rs12101261_T were significantly associated with GD risk(OR=1.35, 95%CI 1.19-1.54, P=5.92×10-6; OR=1.32, 95%CI 1.16-1.50, P=2.22×10-5). Logistic regression identified that rs179247 was an independent susceptibility locus of GD. Serum TRAb concentration showed a significant difference(P=0.015)among rs179247_AA, AG, and GG genotypes. Conclusion rs179247 and rs12101261 in TSHR intron 1 are both associated with GD, and rs179247 may contribute risk to GD independently. The polymorphism is associated with TRAb, but not with serum concentration of thyroid hormones, age of onset, diffused thyroid goiter, ophthalmic signs, and relapse.

Objective To explore the location of the centre of resistance for the maxillary complex in cleft lip and palate by the use of finite element analysis. Methods Combining spiral CT scanning technology with the three-dimensional finite element method, a three-dimensional FEM model of LeFort Ⅰ , Ⅱ , and Ⅲ complex and soft tissue in cleft lip and palate was developed for analysis. Anteriorly and inferiorly directed forces of 9.8N were applied at five different levels parallel to the functional occlusal plane and four different levels perpendicular to the functional occlusal plane, respectively.For each loading condition, horizontal and vertical displacements of different anatomic points in the complex and on the maxillary dentition were analysed. Location of the centre of resistance in different osteotomy complex were studied. Results The resistant center of the LeFort Ⅱ complex in cleft lip and palate was located on intersection between basis nasi and medium of apertura piriforms vertically,apex of the canine and posterior point of the first bicuspid horizontally. The resistant center of the LeFort Ⅲ complex in cleft lip and palate was located on intersection between anterior of the nasion and medium of apertura piriforms vertically, posterior point of the first molar and first bicuspid horizontally. Conclusion Knowledge of the resistant center of different osteotomy complex could establish a basis for biomechanical studies of craniofacial complex distraction osteogenesis in cleft lip and palate.

Objective To explore a method of large cranial bone defect reconstructed by titanium implant with computer aided design(CAD)/computer aided manufacture(CAM)technique.Methods From April 2006 to June 2008,7 cases of cranial bone defect due to tumor and trauma were admitted.The data of skull bone defects were obtained by CT.The resin model was designed and manufactured with rapid prototyping technique.Results The CT data could be used by image software directly.The resin model was manufactured accurately by RP technique.The titanium implant design could be completed by CAD/CAM.7 patients achieved one stage healing.After a follow-up of 6 months to 1 year,cranial bone defect was reconstructed satisfactorily.Conclusion Individual design and repair of large cranial bone defect with CAD/CAM technique is worth extending application clinically.It is a quite ideal and very simple method for the surgical treatment of the cranial bone defect.

The therapeutic effectiveness of nutritional support in the treatment of severe chronic hepatitis and posthepatitic cirrhosis was evaluated. 143 patients with severe chronic hepatitis and 83 with posthepatitic cirrhosis were evaluated with SGA for assessing the nutritional status before the treatment. Patients with severe chronic hepatitis were divided into three groups: group A subject to enteral nutrition (EN) and parenteral nutrition (PN), group B subject to comprehensive treatment (CT)+PN; group C subject to CT+EN. The patients with posthepatitic cirrhosis were divided into two groups: group D receiving CT and group E receiving CT+PN+EN. The function of liver and kidney and nutritional status were monitored to assess the therapy in 6 weeks. The results showed before treatment, over 90 % patients had moderate to severe malnutrition. After nutritional support, the liver function (ALT, T-bil) and nutritional status (TP, TC) in group A was improved significantly as compared with that in groups B and C (P<0.05). Compared with group D, the values of TP and Alb were increased significantly in group E (P<0.05), but the levels of ALT, AST and T-bil had no obvious change. It was suggested that most patients with severe chronic hepatitis or posthepatitic cirrhosis had malnutrition to varying degrees. The nutritional support treatment could obviously improve the nutritional status of these patients, and was helpful to ameliorate the liver function of the patients with severe chronic hepatitis. Among the methods of nutritional support treatment, PN combined with EN had the best effectiveness.

Objective To confirm the diagnosis and to localize the pathogenic gene of ectodermal dysplasia in a family SUffering from only hair and nail abnormalities.MethodsBlood samples were collected from 7 affected patients and 15 unafiected individuals in the family.Genomic DNA was extracted from blood samples by routine phenol-chloroform methods.The whole coding regions of candidate genes K16,K17,K6a,K6b and GJB6 were amplified by PCR followed by direct sequencing.Then,the gene mutation was further confirmed at mRNA level by RT-PCR.ResultsA heterozygous missense mutation 3 1G→A in the GJB6 gene.which leads to the substitution of glycine by arginine at codon 11(G11R)on the N-terminal of the protein,was detected in all the patients.but in none of the 15 normal individuals in this family.The mutation was also confirmed in the CDNA originating from the proband's skin biopsy.Conelusionn A missense mutation G31A.which has been shown previously to cause hidrotic ectodermal dysplasia(HED),is localized in the GJB6 gene of patients in this family.

Objective To investigate the effects of the genetic polymorphisms in osteoporosis-related genes and the gene-gene interaction on bone mineral density (BMD) and osteoporotic fractures.Methods Thirty-nine single nucleotide polymorphism (SNP) sites in 23 genes that related to bone mineral density ( BMD ) and osteoporotic fractures were scanned in 683 Shanghai Han postmenopausal women.TaqMan SNP Genotyping Assay or Sequenom Mass ARRAY System were applied for genotyping analysis.The relation of these SNP sites with BMD and osteoporotic fractures were analyzed.Results Altogether,12 SNPs in 9 candidate genes ( rs7524102 and rs6696981 in ZBTB40 gene,rs9479055 in ESR1 gene,rs6993813,rs6469804,and rs11995824 in OPG gene,rs3736228 in LRP5 gene,rs1107748 in SOST gene,rs87938 in CTNNB1 gene,rs1366594 in MEF2C gene,rs7117858 in SOX6 gene,and rs10048146 in FOXL1 gene) were associated with BMD at lumbar spine(L1-L4) or total hip.In addition,rs11898505 in SPTBN1 gene was related to osteoporotic fractures ( OR 0.522,95％ CI 0.326-0.838,P =0.007 ).Gene-gene interaction involving rs1038304 in ESR1 gene,rs1366594 in MEF2C gene,and rs10048146 in FOXL1 gene was associated with osteoporotic fractures ( P =0.010 7 ).Conclusions ( 1 ) SNPs in gene ZBTB40,ESR1,OPG,LRP5,SOST,CTNNB1,MEF2C,SOX6,FOXL1,and SPTBN1 are associated with BMD of lumbar spine or total hip,as well as osteoporotic fractures.(2) Gene-gene interaction involving rs1038304,rs1366594,and rs10048146may contribute to the risk of osteoporotic fractures.

Objective To investigate the mutations of pedocyte molecules in patients with late onset familial focal segmental glomerular sclerosis (FSGS). Methods Thirty-one pedigrees of late onset familial FSGS in Department of Nephrology, Shanghai Ruijin Hospital from Sep 1997 to Oct 2007 were enrolled in this study. The diagnosis standard of familial FSGS was as follows:(1) the age of presentation was more than 12 years old. (2) in one pedigree, two or more individuals were proven as FSGS by renal biopsy, or at least one was proven to be FSGS by renal biopsy, the others presented renal insufficiency or pmteinuria without precise causes. One hundred unrelated healthy people were screened as control group. Genomic DNA extracted from peripheral blood cells were amplified by PCR and then sequenced for mutations of NPHS2, ACTN4 and TRPC6. Results A novel missense heterozygotic mutation L316P of ACTN4 was identified inone pedigree. The mean onset age of the affected members of this pedigree was (38.7±7.4) years old and their kidney injury progress was slow. Proteinuria of the proband's brother was not improved by immunosuppressor. All 3 affected members of this family had such heterozygotic mutation. A novel missense heterozygotic mutation Q889K of TRPC6 was found in another pedigree. The mean onset age of the affected members in this pedigree was (38.0±4.2) years old. Three members presenting renal disease in this family all had such heterozygotic mutation but with different clinical manifestations. A quiescent mutation G467G of TRPC6 was also identified. Above variants were not found in healthy controls. No NPHS2 mutation was found to cause familial FSGS in these pedigrees. Conclusions A novel mutation L316P of ACTN4 and a new mutation Q889K of TRPC6 are identified in Chinese patients of late onset familial FSGS. No NPHS2 mutation is found to induce FSGS in these pedigrees.