Objective To construct a phage antibody Fab library of human antinuclear antibody. Methods Total RNA was extracted from peripheral blood lymphocytes of 4 patients suffering from autoallergic disease, with antinuclear IgG antibody titers in the serum higher than 1∶10 000 as estimated by indirect immunofluorescence technique. Taking oligo-dT as the primers, human immunoglobulin light chains ?/? genes as well as heavy chains Fd genes were reversely transcribed to cDNA by PCR. The ?/? light chains were initially cloned into pComb3Hss vector to construct a human recombinant light chain library, and the heavy chain genes were subsequently inserted into the corresponding sites of ?/?-pComb3Hss plasmid to generate a recombinant ?/?-Fd-pComb3Hss plasmid. The plasmid was then transformed into E. coli XL1-Blue, which was subsequently infected by the helper phage M13KO7. A random recombinant antibody library was expressed on the surface of filamentous phage. Results Human immunoglobulin ?/? light chain and heavy chain Fd genes (660bp) were amplified successfully. The light chain library with the capacity size of 2?104, and the human recombinant Fab antibody library with the capacity size of 4?104 were also successfully obtained. Finally, the phage antibody library of the Fab fragment of human antinuclear antibody, containing 2?109 CFU/ml phage, was constructed. Conclusion A phage antibody library of Fab fragment of human antinuclear antibody is constructed successfully. This research lays a foundation for the preparation for phage library of Fab fragment of human antinuclear antibody, and also paves the way for the advanced assessment for its effect on the immuno-targeting therapy of tumors.

Objective To study on the effectiveness and reliability of endoscopic varicose vein ligation (EVL) combined with hand-assisted laparoscopic splenectomy (HLS) in portal hypertensive patients. Methods Retrospectively 15 cases of EVL combined with HLS were reviewed from June 2001 to October 2004. These patients with severe esophageal varicose and hypersplenism were adapted to perform endoscopic varicose vein ligation, then 1 -2 weeks after the varicose became milder or disappeared, hand-assisted laparoscopic splenectomy was conducted. Results The number of preoperative ligation was 2. 2 times in average. Esophageal varicose was completely disappeared in 11 and mild in 4. There were no postoperative complications, conversion and death in Hand-assisted laparoscopic splenectomy. The average postoperative count of platelets were (38-67) ? 106/L preoperatively raised to (89-310) ? 106/L postoperatively. In the follow up period (average 17. 6 months) , no varicose vein bleeding happened. Conclusions EVL combined with HLS is not only minimal invasive, but also a secure and effective measure for portal hypertension.

0.05),hints that the tenocyte's function was not disturbed. The DNA index of cells of GDBM group was 0.96 and 2.1% higher than the control group,indicating that the tenocytes grow and proliferate faster when being combine cultured on GDBM. Conclusion GDBM show good biocompatibility combined with tenocytes and they are promising extracellular matrix scaffold for cell transplantation in tendon tissue engineering.

Objective To investigate the clinical effects of gastric bypass( GBP) on type 2 diabetes mel-litus(type 2 diabetes mellitus, T2DM)patients with different body mass index(BMI).Methods T2DM patients undergoing GBP from Sep.2012 to Jul.2013 were divided into 2 groups:obese group( BMI≥28 kg/m2 , 16 ca-ses)and overweight group(BMI<28 kg/m2,21 cases).Changes of fasting plasma glucose(FPG), insulin resist-ance index(HOMA-IR)and insulin stimulation release(INS), C peptide release, BMI, glycosylated hemoglobin ( HbA1c) at 3 and 6 months after surgery were observed.Results INS and C peptide improved 3 months after surgery(P<0.05),and they were close to normal at 6 months after surgery.HbA1c significantly decreased 3 months after surgery compared with that before surgery( P<0.05) and it was close to normal at 6 months after sur-gery.Patients in obese group had significantly declined weight 3 months after surgery compared with that before surgery, and reached the steady state 6 months after surgery.15 cases(93.7%) in the obese group had BMI<25 kg/m2 , and the difference had statistical significance ( P<0.05 ) .Patients in the overweight group had their weight declined but no statistical difference was found(P>0.05).Blood sugar in the obese group decreased with different degrees at 3 and 6 months after surgery.The effective rate was 92%for the obese group and 78%for the overweight group.The overall effective rate was 90%.Conclusion GBP has significant therapeutic value for T2DM patients with different BMI, especially for patients in obese group, which is worth of clinical promotion.

ObjectiveTo study the variant differentiated phase of the human bone marrow stromal cell (MSC) during induced adipogenesis course by morphological observation.MethodsAfter proliferated in vitro, MSCs isolated from bone marrow of the female adults volunteers were cultured with the adipogenetic inducers for 6—30 days. Morphological changes of the cells were observed everyday under Olympus contrast phase microscopy, and some specimens were stained with Oil-red-O.ResultsMSC showed long spindle shape before inducing, and then changed gradually to oval or round shape and the figure enlarged simultaneously. There were specifically morphological changes with lipid droplet emergence under plasmalemma and confluence gradually to lipid drop during differentiated into the phase of immature and mature adipocyte.ConclusionIt is easy to detect the MSC differentiated into the phase of immature and mature adipocyte with the specific morphological change of lipid droplet, while in the phase of preadipocyte and adipoblast, there is no special morphological change, and it may need some specific markers to detect.

Objective To explore the role of sialic acid-binding immunoglobulin-like lectin-one (Siglec-1) in the process of atherosclerotic inflammation induced by oxidized low-density hpepmtein (ox-LDL). Methods Ox-LDL was synthesized by oxidization of native LDL and different concentration of ox-LDL was added to the culture medium of RAW264.7. Forty-eight hours later, cells and supernatants were collected separately. The expression of Siglec-1 protein and mRNA were measured by flow cytometry(FCM) and real-time quantitative RT-PCR, respectively. The levels of monocyte chemotactic protein-1 (MCP-1), macrophage inflammatory protein-1α(MIP-1α) and IL-8 in supernatants were determined by ELISA. Re-sults By the stimulation of ox-LDL, Siglec-1 protein and mRNA on RAW264.7 cells were significantly in-creased, meanwhile, the cytokines levels in culture supematants were significantly higher than that in the control group. And both Siglec-1 expression and cytokine secretion were ox-LDL dose-dependent. Conclu-sion Ox-LDL can increase Siglec-1 protein and mRNA expression and some inflammatory cytokines secre-tion on RAW264.7 cells in a dose-dependent manner. Manifested by enhanced Siglec-1 expression, the acti-vated macmphages may take a part in the development and progression of atherosclerosis.

Objective To investigate the effect of combined double-stranded RNA (dsRNA) and imiquimod stimulation on uterine immune cells. Methods In BALB/c × C57BL/6 mice and non-obese dia-betic (NOD) × C57BI/6 mice, embryo resorption rate was detected in the presence or absence of Toll-like receptor 3 (TLR3) agonist dsRNA [poly( 1: C)], TLR7 agonist imiquimod ( R837), or their combination, respectively. In in vivo system, the status of intracellular cytokine production in uterine CD45 + cells was de-tected by flow cytometry. To identify the CD45 + cells, uterine CD3+ T cells and CD49b + NK cells derived from placenta and decidua basalis were stimulated with dsRNA and imiquimod in in vitro systems, and the status of intracellular cytokine production was detected. Mitogen-activated protein kinase (MAPK) antago- nists SP600125 and PD98059 were used to block the increase of cytokine production. Results A synergistic increase of embryo resorption was observed after the induction of dsRNA and imiquimod combination. Mean-while, a synergistic increase of TNF-α and IFN-γ production was detected after the induction in CD45 + cells. Further study found that although synergistic effect can be detected in both CD3 + cells and CD49b + cells in BALB/c mice, the status was different in NOD mice. The cytokine increase should mainly be attrib-uted to CD3 + T cells, since no such increase was detected among the CD49b + NK cells in the NOD mice. The synergistic effect of combined agonists was partially inhibited by Jun N-terminal kinase (JNK) MAPK inhibitor SP600125 and almost completely abrogated by extracellular signal-regulated kinase (ERK) MAPK inhibitor PD98059. Conclusion Boosted TLR3 and TLR7 signal may be transmitted via Thl-type T cells, rather than NK cells in NOD mice. ERK MAPK pathway may be critical in TLR3 and TLR7 involved signa- ling.

Objective Comparing the difference in postoperative recurrence,complications,quality of life and cosmetic results between patients receiving radical mnastectomy and breast conserving surgery,provides an evidence of breast conserving surgery superior to radical mastectomy.Methods A retrospective analysis of 477 breast cancer patients cases in Department of General Surgery,Third People's Hospital of Baoji City from January 2009 to January 2012.These patients were divided into two groups:the control group 229 cases (48％) underwent conservative surgery treatment and the observation group 248 patients (52％) underwent radical surgery.Using SPSS15.0 statistical software analysis and compare with recurrence,postoperative complications,breast cosmetic effect and quality of life for these two groups of patients.Results In breast-conserving group compared with radical mastectomy group,the one and two year recurrence or metastasis rate were not statistically different between the two groups (P ＞ 0.05),the incidence of postoperative complications was significantly decreased (P ＜ 0.05).The scores for quality of life between the breast-conserving group and radical mastectomy group were all significantly different (P ＜ 0.05).Excellent cosmetic results in breast-conserving group was 78.52％,which was significantly higher than that in the radical mastectomy group (61.34％),the difference was statistically significant (x2 =5.86,P ＜ 0.05),The two groups are not significant in overall survival time (x2 =3.154,P ＞ 0.05) and progression free survival (x2 =4.243,P ＞ 0.05) as two indicator of long-term efficacy.Conclusions Conservative surgery compared with radical mastectomyhave less clinical complications,more breast cosmetic effect,better survival quality,and both of them share the same recurrence or metastasis and survival rate,so conservative surgery should be preferable in the clinical application.

This study was conducted to investigate the biomechannical properties of tissue-engineering cartilage. The compressive modulus of the collagen-chondrocyte constructs that had been cultured in vivo for 16 weeks and the centrifuge-tube-cultured cartilage in vitro were measured with 5% strain. The results showed that the compressive moduli of the centrifuge-tube-cultured cartilage at the 4th, 8th, 12th and 16th weeks were 0.352MPa, 0.653MPa,0.233MPa and 0.262MPa respectively. The top modulus among them was at the 8th week. The compressive modulus of the neocartilage engineered by the collagen-chondrocyte constructs in vivo for 16 weeks was 10.668 MPa and reached the same order of magnitude as human fetal articular cartilage, but it is still under that of the human fetal articular cartilage actually. The culture methods should be developed further though the dynamic alteration of the compressive modulus of the centrifuge-tube-cultured neocartilage is in accord with that of the GAG/DNA content ratio of the neocartilage.

To detect the properties of natural xenogeneic bone derived materials which were processed with different physical and chemical treatments, we made fully deproteinized bone(FDB), partially deproteinized bone (PDPB), partially decalcified bone(PDCB) from pig ribs. Their morphological features, constitute components and mechanical properties were examined by scanning electron microscopy, x-rays diffraction analysis, mechanical assay and so on. The results showed that FDB, PDPB and PDCB maintained natural network pore system. The ratios of calcium to phosphorus were 1.81, 1.74 and 1.50, and the protein contents were 0.01% +/- 0.02%, 22.41% +/- 0.83% and 35.75% +/- 2.12% respectively. The sequence of their mechanic strength was PDCB > PDPB > FDB. These data indicate that FDB, PDPB and PDCB possess natural network pore system. Their organic and inorganic component ratios and contents are different, so their mechanic properties are not alike. Additionally, more investigations will be necessary to detect the biocompatibility of the three different scaffold materials of natural derived bone.
Animals ; Biocompatible Materials ; chemistry ; Biomechanical Phenomena ; Bone and Bones ; chemistry ; Materials Testing ; Swine ; Tissue Engineering