Objective To investigate the effects of c?myc promoter binding protein 1(MBP?1)gene on the proliferation of human Saos?2 osteo?sarcoma cells in vitro. Methods Saos?2 cells were divided into three groups:blank control group(untransfected cells),negative group(cells transfected with missense sequence)and experimental group(cells transfected with MBP?1 shRNA). Two MBP?1 shRNA sequences and one neg?ative control shRNA sequence were designed ,synthesized and cloned into pSIREN?retroQ plasma. Then the recombinant plasmids were construct?ed and transfected into human Saos?2 osteosarcoma cells by Lipofectamine 2000. The expressions of MBP?1 mRNA and protein in Saos?2 cells were detected by real?time PCR and Western blot ,respectively. The effects of altered expression of MBP?1 on cell proliferation were measured by CCK?8 cell proliferation assay. The expressions of cyclin D1 and cyclin E in Saos?2 were determined by Western blot. Results PCR and sequenc?ing results indicated that the recombinant plasmids pSIREN?retroQ was constructed. The relative expression level of MBP?1 mRNA in the MBP?1 siRNA transfection group was significantly decreased than that in blank control group(P<0.05). Compared with the blank control group,the ex?pression levels of MBP?1 protein in the experimental group also significantly decreased. The proliferation abilities of Saos?2 cells at 48,72,and 96 hours after MBP?1 siRNA transfection were significantly increased than those in the blank control group(P<0.05). Compared with the blank con?trol group,the expression levels of cyclin D1 and cyclin E protein in the experimental group also significantly increased(P<0.05). Conclusion Knockdown of the expression of MBP?1 gene promotes the proliferation of human Saos?2 osteosarcoma cells. MBP?1 gene may become the new tar?get of gene therapy for osteosarcoma.

A review of cell traction forces (CTFs) measurement based on Biological MiCro Electromechanical Systems (BioMEMS) microposts matrix is presented.CTFs are exerted by cells and ansmitted to the underly-ing substrate through focal adhesions and close contacts.which is essential for cells movement.Cells probe the mechanicaI compliance of the exlracellular mabix (ECM) in part by locally deforming it with nanonewton-scale traction forces.Precision measurement of CTFs is significant for many researches such as call biology and tissue engineering and so on.Enabled by the advancement in BioMEMS technology,surface treated high aspeect ratio Polydimethyisiloxane(PDMS)micropos matrix devices,which serve as BioMEMS sensom for de-tecting cellular nanoforces and studying in vitro cell mechanics,have been developed.Closely spaced vartical microposts matrixes were designed to encourage cells to attach and spread across multiple microposts,and to bend the microposts like vertical cantilevers as the cells locomote on the surface.Using this dense and dis-crete matrix of microposts rather than a convanfional continuous substrate,CTFs can be directly measured and quantified by processing the microscopy images of the deformations of microposts.The resolution of the force was in tens of nN/μm scale.At first,the conventional CTFs measurement methods were concisely summa-rized.Then BioMEMS microposts matrix method was described in detail,including principle and fabfication process,Surface treatment and cell expedment results.Furthermore,high aspect ratio structure collapse prob-lem was investigated.

A review of cell traction forces (CTFs) measurement based on Biological MiCro Electromechanical Systems (BioMEMS) microposts matrix is presented.CTFs are exerted by cells and ansmitted to the underly-ing substrate through focal adhesions and close contacts.which is essential for cells movement.Cells probe the mechanicaI compliance of the exlracellular mabix (ECM) in part by locally deforming it with nanonewton-scale traction forces.Precision measurement of CTFs is significant for many researches such as call biology and tissue engineering and so on.Enabled by the advancement in BioMEMS technology,surface treated high aspeect ratio Polydimethyisiloxane(PDMS)micropos matrix devices,which serve as BioMEMS sensom for de-tecting cellular nanoforces and studying in vitro cell mechanics,have been developed.Closely spaced vartical microposts matrixes were designed to encourage cells to attach and spread across multiple microposts,and to bend the microposts like vertical cantilevers as the cells locomote on the surface.Using this dense and dis-crete matrix of microposts rather than a convanfional continuous substrate,CTFs can be directly measured and quantified by processing the microscopy images of the deformations of microposts.The resolution of the force was in tens of nN/μm scale.At first,the conventional CTFs measurement methods were concisely summa-rized.Then BioMEMS microposts matrix method was described in detail,including principle and fabfication process,Surface treatment and cell expedment results.Furthermore,high aspect ratio structure collapse prob-lem was investigated.

Objective To evaluate the nutritional status of iodine of pregnant and lactating women in coastal and inland areas of Zhejiang province,and to provide scientific basis for iodine supplementation.Methods In 2009,five coastal cities,five coastal counties and three inland areas from Zhejiang province were selected by the cluster sampling method,and one sub-district(township) was randomly selected in each city (county),then one community (village) was randomly selected in each sub-district (township).Thirty pregnant women,thirty lactating women and fifty children aged 8 - 10,were randomly selected in each community(village) as the investigation objects to collect instant urine samples to detect urine iodine,and to collect thirty resident edible salt and some water samples to detect salt iodine and water iodine,if the numbers of water source were more than 5 then 1 water sample was collected by their location of east,south,west,north and center in each community(village),otherwise,the numbers of water samples collected were equal to the number of community (village).Results The medians of salt iodine of coastal city,coastal county and inland area were 28.42,25.29,and 33.24 mg/kg,respectively,and the medians of water iodine correspondingly were 5.41,11.52,and 2.80 μg/L,respectively.The medians of urinary iodine of pregnant and lactating women in Zhejiang province were 148.72 and 161.34 μg/L,respectively,which were less than that of children aged 8 - 10(231.02 μg/L,Z =8.57,8.24,all P ＜ 0.017).The medians of urinary iodine in pregnant women from different areas were ranked in order as inland areas(211.42 μg/L),coastal counties (150.51 μg/L) and coastal cities (123.33 μg/L,Z =2.80,5.31,2.42,all P ＜ 0.017),and in lactating women,the medians of urinary iodine in inland areas( 197.43 μg/L) was higher than that in coastal cities( 139.64 μg/L,Z =4.03,P ＜ 0.017).Conclusions The overall levels of iodine nutrition in lactating and pregnant women in Zhejiang province are in the appropriate level,but that of the pregnant women in the coastal cities is inadequate.

Objective To investigate the blood supply of the expanded skin flap from medial up-per arm and its application in the reparation of cervical scar contracture due to sear resection. Methods The operation was carried out for three steps: (1) The expander was implanted under the superficial fascia. (2) The skin flap from medial upper arm was created with superior ulnar collateral artery as blood supply and attributive branches of basilica and axillary veins as blood collection. (3) After thes car contracture was released, the defect was covered with medial upper arm flap with maximal area of 25 cm×15 cm. Results Ten patients in all with cervical scar contracture were treated with the skin flap. All the skin flaps survived at last with nearly normal skin color, texture and contour. And the scar in donor sites seemed to be neglectable. Conclusions Reparation of cervical scar contracture with medial upper arm skin flap after expanding could be recommended. But 3 months long time and fixation of upper limb and head might be disadvantages.

OBJECTIVETo explore the activation and proliferation of specific T cells induced by artificial antigen-presenting cells (aAPCs) simulated dendritic cells (DCs) and to observed the effect of these T cells on leukemic cell killing.

METHODSaAPCs were developed by coating a human leukocyte antigen-immunoglobulin fusion protein ( HLA-lg), which was connected each one of the four CML28 antigen epitopes (DLMSSTKGL, DLMSSTKGL, ALFCGVACA, VLTFALDSV), and CD28-specific antibody, to magnet-beads CML cell specific peptides (CML28) served as target peptides. Bone marrow (BM) or peripheral blood (PB) mononuclear cells (MNCs) were isolated from HLA-A2 healthy volunteers, and co-cultured with aAPCs. Specific T lymphocyte were detected by flow cytometry. The fresh acute leukemic cells were used as target cells. The specific T cells incubated with leukemic cells for 4 h at ratios of 5:1, 10:1, 20:1, 40:1, 80: 1, respectively. The effect of leukemic cells killing was detected by lactate dehydrogenase release test.

RESULTSThe average ratio of CML-28 specific T lymphocyte in control group was (2.2 +/- 0.4)% and in experimental groups (DLMSSTKGL, DLMSSTKGL, ALFCGVACA, VLTFALDSV) were (13.5 +/- 1.6)%, (15.2 +/- 1.5)%, (14.7 +/- 1.8)% and (34.3 +/- 3.5)%, respectively, being significantly higher than that in control group (P < 0.01). Induction efficiencies of acute leukemic cells killing were significantly enhanced by increase of effector cells. The cytotoxic activity of specific T lymphocyte in one experimental group (VLTFALDSV) was much higher than that in other three experimental group (P < 0.05).

CONCLUSIONThis "prime and expand" regimen should be an alternative method for large scale amplification of rare tumor-specific CTLs and aAPCs might be a useful tool for leukemia immunotherapy.

Adolescent ; Adult ; Antigen-Presenting Cells ; immunology ; Cell Proliferation ; Cells, Cultured ; Coculture Techniques ; Cytotoxicity, Immunologic ; Female ; Humans ; Leukemia ; immunology ; pathology ; Lymphocyte Activation ; Male ; Middle Aged ; T-Lymphocytes ; cytology ; immunology ; Young Adult

OBJECTIVETo compare differences between Crowe IV developmental dysplasia of the hip (DDH) with secondary acetabulum and Crowe IV DDH without secondary acetabulum,and determine whether it is necessary to divide Crowe IV DDH into two subtypes.

METHODSFrom June 2007 to May 2015,145 hips of 112 Crowe N patients who underwent total hip arthroplasty (THA) using S-ROM stem were divided into two groups: secondary acetabulum formaton group (group A) and no secondary acetabulum formaton group (group B). In group A,there were 12 females, 96 males,with an average age of (39.38 ± 11.19) years old. In group B, there were 2 females, 35 males, with an average age of (38.19 ± 10.92) years old. All the patients were evaluated by using Harris Hip Score. Radiographic evaluations were made preoperatively and during follow up. The differences between two groups were compared on dislocation height, canal flare index (CFI), subtrochanteric shortening osteotomy (SSTO) usage, pre- and post-operation Harris scores, complications.

RESULTSThe dislocation height for group A was (4.74 ± 1.57) cm, while the dislocation height for group B was (3.12 ± 1.15) cm. Significantly difference was detected between two groups. The CFI for group A was 2.69 ± 0.68, while the CFI for group B was 3.42 ± 0.79, and the significantly difference was detected between two groups. Harris scores were totally improved from 58.18 ± 15.67 preoperatively to 91.20 ± 3.79 post-operatively and the difference was significant. Pre-operative Harris scores was 58.1 ± 15.3 in group A, 58.3 ± 16.9 in group B. Post-operative Harris scores was 91.0 ± 4.1 in group A, 91.0 ± 5.1 in group B. No significant difference was found on Harris scores between A and B preoperatively and post-operatively. Complications of 4 cases peri-prosthesis fracture, 4 cases dislocation and 4 cases nerve injury occur in group A; While only one case dislocation and one case nerve injury occur in group B. No statistical significance was detected.

CONCLUSIONCrowe IV DDH with secondary acetabulum is significantly different from Crowe IV DDH without secondary acetabulum on dislocation height and femoral morphology, which causes the different selections of surgical techniques (SSTO usage or not). These important differences in fundamental parameters indicate the necessity to further divide Crowe IV DDH into IVA and IVB two subtypes.

Adolescent ; Adult ; Aged ; Female ; Hip Dislocation, Congenital ; classification ; surgery ; Humans ; Male ; Middle Aged ; Postoperative Complications ; therapy

Objective To explore the microanatomic features of the transcallosal-interforniceal approach, and discuss the value of its clinical application. Methods Fifteen adult cadaveric head specimens were dissected by microsurgical anatomic skills to simulate the procedures of the transcallosal-interforniceal approach. Observation and measurement were performed on related anatomic structures. In clinical, 21 patients with the third ventricle tumors underwent tumor resection via the transcallosal-interforniceal approach. Results Using two points on the cortical surface as references that were located 5 and 7 cm anterior to the central sulcus respectively, mean values of related measurements on P5-foramen of Monro (FM) and P7-FM were obtained as follows: (1) the depth of the interhemispheric fissure was 38.46 and 37.62 ram; (2) the height of the corpus callosum was 7.18 and 7.78 nun; (3) the height of the septum pellucidum was 7.53 and 9.88 mm; (4) the thickness of the fornix was 4.72 and 5.16 mm. Under the operative microscope, the tumors were totally removed in 11 cases,subtotally in 8 cases, and partially in 2 cases. Conclusions The corridor of the transcallosal-interforniceal approach should be limited between the lines of P5-FM and P7-FM. The quantitative information obtained in this study permits the preservation of important anatomic structures in operation, such as the motor strip, genu of the corpus callosum, fornical commissure and anterior commissure. This approach is deserved to be applied generally for providing a quite large operative field,making total tumor removal easier, and decreasing the incidence of postoperative complications.

OBJECTIVETo study the prevalence of hepatitis C virus (HCV) infection and characteristics on molecular biology related to HCV among patients who were enrolled in a Methadone maintenance clinic in Wuhan.

METHODSSerum samples from 332 injection drug users (IDUs) were obtained and anti-HCV IgG was detected by enzyme linked immunosorbrent assay(ELISA), together with 86 anti-HCV positive specimens genotyped. A reverse transcriptase-polymerase chain reaction (RT-nPCR) assay using conserved primers deduced from the core-envelopel (C-E1) region of the HCV genome was employed to amplify a 474 bp fragment. Phylogenetic analysis of the C-E1 sequences was conducted by direct sequencing of the RT-nPCR products and alignment with determined by nucleotide sequencing followed by composition of a phylogenetic tree.

RESULTSThere were 313 cases (94.3%) appeared positive anti-HCV IgG in the 332 patients from a Methadone maintenance clinic in Wuhan. It was demonstrated that there were four different subtypes of HCV in that clinic in Wuhan, including 6a--71 cases (82.5%), 3b--7 cases (8.2%), 1a--5 cases (5.8%) and 1b--3 cases (3.5%).

CONCLUSIONInfection of 6a genotype HCV was predominant in patients from the Methadone maintenance clinic in Wuhan, followed by HCV 3b, 1a and 1b.

Adult ; Antibodies, Viral ; analysis ; China ; Enzyme-Linked Immunosorbent Assay ; Female ; Genotype ; Hepacivirus ; classification ; genetics ; Humans ; Male ; Methadone ; therapeutic use ; Middle Aged ; Phylogeny ; Reverse Transcriptase Polymerase Chain Reaction ; Substance Abuse Treatment Centers ; Substance-Related Disorders ; drug therapy ; rehabilitation ; Young Adult

Objective To achieve detailed genomic characterization and investigate the antibiotic-resistant mechanisms of plasmid pA1137 carrying the aminoglycoside resistance gene aacC2.Methods Antibiotic-resistant genes were deter-mined by PCR.Conjugation experiments were performed to verify the transferability of plasmid pA 1137.The minimum in-hibitory concentration(MIC)values of bacterial strains were tested with microdilution method.The genetic background, mobile elements and antibiotic resistance mechanisms of pA 1137 were determined using a whole genome sequencing meth-od.Results Both carbapenem-resistant gene blaIMP-8and aminoglycoside-resistant genes aacC2 and aacA4 were carried by A1137 isolated from Enterobacter cloacae(ECL).aacC2 was located in plasmid pA1137 while the other two resistant genes were observed in chromosomes.Plasmid pA1137 was an IncFⅡplasmid,whose total length was 68.97 kb,and GenBank accession number was MF190369.Plasmid pA1137 contained multiple replicons and intact conjugative transfer regions,so it could be transferred into ECL through conjugation experiments and confer corresponding antibiotic resistance to the transconjugant A1137-EC600.Conclusion IncFⅡ plasmid pA1137 has a single accessory region, the first reported Tn5403-based aacC2-tmrB-related region,which can cause stable inheritance and mediate the resistance to aminoglycoside antibiotics in ECL A1137.