In order to construct a single chain fragment variable (ScFv) phage display library against ovarian tumor, by using RT-PCR, the human heavy chain variable region genes (VH) and light chain variable region genes (VL) were amplified from lymphocytes of ovarian tumor patients and subsequently assembled into ScFv genes by SOE. The resulting ScFv genes were electrotransformed into E. coli TG1 and amplified with the co-infection of helper phage M13KO7 to obtain phage display library. The capacity and titer of the resulting library were detected. The phage antibody library with a capacity of approximately 3 x 10(9) cfu/microg was obtained. After amplification with helper phage, the titer of antibody library reached 5 x 10(12) cfu/mL. Human ScFv library against ovarian tumor was constructed successfully, which laid a foundation for the screening of ovarian tumor specific ScFv for the radioimmunoimaging diagnosis of ovarian tumor.

Objective To analyze the genetic diversity of eight species of Caragana Fabr. in Ordos Plateau. Methods To study the genetic diversity of the eight species with ISSR method. Results The total percentage of polymorphic bands (PPB) of the eight species was up to 100%, which was extremely high and showed an abundant genetic diversity. However, the genetic diversity of different species was not the same. The genetic diversity of C. tibetica, C. korshinskii, C. stenophylla, C. roborovskyi, and C. intermedia was higher, while that of C. opulens and C. purdomii was lower, and that of C. brachypoda was the lowest. The analytic results of the Nei′s gene diversities and the Shannon′s information index were similar. Different species could be differentiated through UPGMA method and characteristic banding pattern of ISSR. Conclusion ISSR Molecular marker could be used in molecular authentication and hereditary background study of the species of Caragana Fabr. These results provide the scientific basis of developing effective protection measures for these species.

Objective To study the clinical and genetic features of familial paroxysmal kinesigenic dyskinesia (PKD). Methods The clinical information of 14 family members in one pedigree, including 2 patients (one treated in hos?pital, the other not treated) were analyzed and the response to treatment and prediction were followed up. DNA was ex?tracted from peripheral blood samples, and then screened for PRRT2 mutations. Results There were two male patients in the pure PKD pedigree, Prevalence rate was 14.3%,One of the PKD patients showed good response to carbamazepine as well as lamotrigine whereas other patients recovered without treatment. We detected a nonsense mutation c.797G>A (p.266R>Q) in PRRT2 gene in three family members. One affected member harboring PRRT2 mutation resulted from the incomplete penetrance of the disease,PKD and polycystic kidney disease coexist in the pedigree which showed autoso?mal dominant inheritance with incomplete penetrance and anticipation. Conclusions The curative effect of antiepileptic drugs to purely familial PKD is related to mutations and clinical features;Treatments should be decided based upon clini?cal features and mutations.

The polyclonal antibodies against VLDL receptor were prepared and identified. Rabbits were immunized with polypeptide fragment of VLDL receptor as antigen. The collected blood serum of the immunized rabbits was analyzed and identified by using ELISA and Western Blot. The results showed that the rabbit against mouse and human VLDL receptor antibodies were obtained with high titer and could recognize the natural VLDL receptors through Western blot. The prepared polyclonal antibodies against VLDL receptor provide a new tool to study the protein of VLDL receptor.
Antibodies/chemistry ; Antibodies/*immunology ; Enzyme-Linked Immunosorbent Assay ; Peptides/*immunology ; Receptors, LDL/*immunology

To evaluate the clinical therapeutic effect of the autologous bone marrow mesenchymal stem cells grafting integrating blood plasma transplantation in treatment of problematic nonunion. In March 2000, a 29-year-old man presented complaints of painful walking of left thigh after a velocity injury, was selected. Radiography revealed a left femoral shaft fracture, and the bone defect distance was 5 mm through X-ray examination before operation. Following a series of reposition, fixation, intramedullary nail fixation and twice autogenous lilac bone graft treatment, external fixator and autogenous lilac bone graft treatment, totally four times, the fracture was not healed. He come to Orthopaedic Surgery Department of Siping Central Hospital to accept the autologous bone marrow mesenchymal stem cells grafting integrating blood plasma transplantation in March 2008. Under the small C-arm X-ray perspective, 4 mL bone marrow-derived mesenchymal stem cells suspension integrating 10% autologous blood plasma was injected vertically into fracture site from the skin in front of the thigh with epidural needle, and the stem cells density was 1.8 × 107 cells/L. X-ray examination was performed though out-patient recheck to observe fracture healing. Two months after graft, radiography results showed fracture interspace reduced, left femoral shaft callus was continual and fracture lines blurred partly of the left femur; furthermore, left femoral shaft fracture lines blurred, and continuous bone callus formation of the left femur at 4 months after graft; The left femur achieved bone union within 7 months. The patient was returned to full weight bearing walking and good function with a fully healed femoral bone, without any fever or infection. Percutaneous transplantation of autologous bone marrow stem cells for treatment of problematic nonunion has the satisfactory result.

Adult ; Diagnosis, Differential ; Endometriosis ; pathology ; surgery ; Female ; Follow-Up Studies ; Humans ; Intervertebral Disc Displacement ; pathology ; Lumbar Vertebrae ; pathology ; Magnetic Resonance Imaging ; Spinal Canal ; pathology ; surgery ; Spinal Diseases ; pathology ; surgery

Forty five patients with acute cerebral infarction were randomized to two groups: in treatment group patients received local subhypothermia and conventional therapy, in control group patients received conventional therapy only. Clinical outcome was assessed by the National Institutes of Health Stroke Scale (NIHSS) on admission and at 7, 14 and 30 d after treatment. Serum neuron specific enolase (NSE), nitrogen monoxide ( NO ) , superoxide dismutase (SOD), interleukin-6 (IL-6 ) and intercellular adhesion molecule-1 (ICAM-1) were detected on admission and at 7,14 d after treatment The study showed that NIHSS scores of treatment group on 14, 30 d were lower than those of control group ( P ＜ 0. 05 ). Serum NSE, NO, IL-6 and ICAM-1 levels significantly decrease; while serum SOD levels increased (P ＜ 0. 05). In conclusion, local subhypothermia therapy can inhibit inflammatory reaction, reduce oxygen free radical formation and improve neurological function in patients with acute cerebral infarction.

Objective To study inflammatory reaction induced by N-protein of severe acute respiratory syndrome(SARS)-coronavirus(CoV)in human alveolar typeⅡepithelial cell(A549). Methods Effects on growth of A549 cell by N-protein of SARS-CoV:activity of A549 cells was determined by thiazylyl blue colorimetry assay at 24,48,72 and 96 h,respectively.Effects on cyto- kine production by A549 cells exposed to N-protein of SARS-CoV:interleukin(IL)-6,IL-10 and transforming growth factor-?1(TGF-?1)concentration in culture supernatant were determined by enzyme-linked immunosorbent assay(ELISA).Effects on mRNA expression of cytokine of A549 cells and matrix metalloproteinases-9(MMP-9)exposed to N-protein of SARS-CoV:total RNA of A549 cells was extracted using Rneasy mini kit;RT-PCR was employed to measure the mRNA expression of IL-6,IL-10,TGF-?1 and MMP-9 semiquantitatively.Results Different concentrations of N-protein could all inhibit the growth of A549 cells(after 48 h)and the inhibition by 20?g/mL pro- tein was the strongest.Compared with the control group(0.737?0.024,0.968?0.007),the A val- ues of experimental groups at 72 h and 96 h(0.672?0.027,0.799?0.092)decreased obviously (P

Aged ; Breast Neoplasms ; metabolism ; pathology ; surgery ; Cadherins ; metabolism ; Carcinoma, Merkel Cell ; metabolism ; pathology ; Carcinoma, Small Cell ; metabolism ; pathology ; surgery ; Diagnosis, Differential ; Female ; Humans ; Lymphoma ; metabolism ; pathology ; Melanoma ; metabolism ; pathology ; Phosphopyruvate Hydratase ; metabolism ; Synaptophysin ; metabolism

In order to construct the expression recombinant of human receptor associated protein (RAP), optimize its expression condition and obtain the recombinant protein after expression with high efficiency, two prokaryotic expression vectors-pT7-PL and pET-28a(+) were used to construct the expression recombinant containing RAP cDNA, and the expression efficiency of two kinds of expression E. coli of BL21 strains was compared. The effect of different induction conditions on the expression of recombinant RAP was observed. After recombinant protein was purified with Ni(+) -nitrilotriacetic acid (Ni(+) -NTA) affinity chromatogram, its binding ability with microphage was observed. The results showed that two recombinant plasmids both obtained high expression of RAP. The expression levels of RAP in plasmid pT7-PL-RAP in BL21 (DE3, plysS) strain were significantly higher than in BL21 (DE3) strain. The expression of pT7-PL-RAP in the presence of chloramphenicol was higher than in the absence of chloramphenicol, and most of the inducible expressed RAP was soluble. The RAP which was purified by Ni(+) -NTA resin could strongly bind with the RAW264.7 cells rich in low density lipoprotein receptor (LDLR) family receptors. It was concluded that the expression condition of recombinant RAP was optimized and functional RAP was obtained, which offered a good foundation for the further production of RAP as research tool.