Objective To realize the construction and expression of human dsFv genes to rabies virus. Methods The genes of dsFv VH and VL were amplified by PCR-based point mutagenesis strategy. Then the genes were cloned into plasmid pET-22b (+). E. coli BL21(DE3) was transformed with the recombinant expression plasmid and the protein was induced in Luris-Bertani medium by addition of IPTG. The inclusion body proteins of VH and VL were denaturalized by GuHCl, and then re-naturalized in refolding solution to form dsFv fragments. Afterwards, we evaluated the antigen-binding activity of the dsFv and its stability as compared with its originative scFv. Results The fragment genes of dsFv to rabies virus were constructed successfully by PCR-based point mutagenesis strategy. Sequence analysis proved that cysteines were introduced into the position 44 aa of VH and position 100 aa of VL. The dsFv VH and VL genes were expressed in PET22b(+)/BL21(DE3). The VH and VL protein folded into the active dsFv antibody fragments which showed specific binding capability to rabies virus. Conclusion We succeeded in achieving the stabilization of the human scFv to rabies virus and obtaining the active human dsFv antibody fragments. This established a solid basis for further study of the biological activaty and clinical application of dsFv to rabies virus.

OBJECTIVE:To effectuate the pharmacy information communication with in the hospital.METHODS:The local area network concerning medicine information in the hospital was built by adopting FrontPage and Dreamweaver with the assists of Flash,Photoshop and Fireworks so as to provide services for the communication of pharmaceutical sciences informa-tion and the information collecting.RESULTS:The website on medicine information has been operating normally for11months and the visitors to which were more than5000person-time.CONCLUSION:Medicine information website has provided an active,healthy and an upward platform of communication and learning for all medical workers within the local area network of the hospital,which has make the gathering of clinical pharmacy information more promptly and effectively and which has achieve the expected effect.

Objective To analyze the clinical and pathological features of limb-girdle muscular dystrophy2B(LGMD2B).Methods The clinical and pathological features of 7 patients with LGMD2B were analyzed retrospectively.Results Seven patients had a slow onset,and progressive proximal muscle weakness,muscle atrophy,progressive,and incresed serum creatine phosphokinase; muscle biopsy showed different degree of muscle fiber degeneration,necrosis; stromal and inflammatory cell infiltration in muscle fiber; monoclonal antibody immunohistoehemical staining:showed expression of Dysferlin protein was not found in muscle cell membrane,Dystrophin,Sarcoglycans protein expression was normal.Monoclonal antibody immunohistochemical staining the proteins were expressed in normal cell membrane.Conclusions LGMD2B is a slow onset,progressive proximal muscle weakness,muscle atrophy.Histochemical staining on the basis of further immunohistochemical staining,to detect the membrane protein and Dysferlin protein expression,that is a necessary means to diagnose LGMD2B and inflammatory myopathies.

Objective To identify the problems of medical record management standardization in clinical case management for the study of medical record quality and give some corresponding measures.Methods 3,000 medical records of patients in our hospital from January 2015 to December 2016 were collected and divided into control group (with 1,500 records from January 2015 to December 2015) and observation group (with 1,500 clinical medical records from January 2016 to December 2016).The control group adopted regular data management while the observation group employed standardized lean management.Medical record defects, rectification rate and the average scores of inpatient medical records between the two groups were compared and analyzed.Results The average score of inpatient medical records in observation group was significantly higher than that of the control group (t=13.672, P<0.05), while the defect degree and the modification rate of the medical records were significantly lower than those of the control group (P<0.05).Conclusion Standardized medical record management contributes to the improvement of the quality of medical records.

Objective To observe the effect of Ulinastatin (UTI) on intestinal barrier function of elderly patients with sepsis in order to explore an optimized program of early treatment for such patients instituting a guidelines for clinical practice.Methods All selected patients randomly (ramdom number) divided into conventional treatment group (n =34) and UTI treatment group (n =34).Peripheral venous blood samples of elderly patients with sepsis were routinely taken at days 0,2,3,7 (D0,D2,D3,D7) after admission for detecting the changes of ptasma values of Intestinal fatty acid binding protein (IFABP),Diamine oxidase (DAO),and D-lactate (D-Lac).While patients' APACHE Ⅱscores,length of ICU stay and 28 d mortality also were recorded at the same time.Results At admission,there were no significant differences in patients' APACHE Ⅱscores,plasma values of IFABP,DAO and D-Lac between two groups (P ＞ 0.05).During the course of treatment,the patients' APACHE Ⅱscores,plasma values of IFABP,DAO and D-Lac were declining in both groups,while those declinations in UTI group showed significantly more prominent.Compared with conventional therapy group,IFABP level was significandy lower in UTI group (P ＜ 0.05),and the plasma values of DAO and D-Lac were significantly lower at D2 and D3 in UTI group (P ＜ 0.05),but not at D7 (P ＞ 0.05).The plasma levels of IFABP,DAO and D-Lac were positively associated with APACHE Ⅱscores,but not with length of ICU-stay (P ＞0.05).After treatment,28d mortality did not differ significantly between the two groups (P ＞0.05).Conclusions UTI has a protective effect on intestinal barrier function in elderly patients with sepsis.UTI can improve the prognosis of elderly patients with sepsis and reduce ICU stay,while has no significant effect on reducing 28 d mortality.

Objective To explore the relationship between the damage of neuromyelitis optica (NMO) astrocytes (AS) and the onset of NMO, and investigate the relevance of AS damage with the severity of the patients with functional defect. Methods The levels of aquaprin4-antibody (AQP4 -Ab), glial fibrillary acidic protein (GFAP), apolipoprotein(ApoE),interleukins-6(IL-6), interleukins-10(IL-10), tumor necrosis factor-α(TNF-α) in cerebrospinal fluid (CSF ) and serum of 30 acute NMO patients were tested by means of ELISA. The results were later compared with control group. And analysis of the relevance of the various index of the levels in CSF with the CSF AQP4-Ab level, the acute phase expanded disability status scale(EDSS) score of the NMO group were made. Results (1)The NMO group in CSF and serum AQP4-Ab, GFAP, IL-6 levels were higher than the control group (P < 0.05), and ApoE, IL-10 levels were lower than the control group (P < 0.05). (2)The CSF GFAP, ApoE, IL-6 in NMO group is higher than the serum (P < 0.05), and CSF AQP4-Ab, IL-10 levels were lower than the serum ( P < 0 . 05 ) . ( 3 ) The CSF GFAP , IL-6 levels and the CSF AQP4-Ab level were positively correlated (r=0.749, r=0.526, P<0.05), and the CSF ApoE, IL-10 levels were negatively correlated with CSF AQP4-Ab level(r = -0.571, r = -0.676, P < 0.05). (4)The CSF AQP4-Ab, GFAP, IL-6 levels and the acute phase EDSS score were positively correlated (P < 0.05), the CSF ApoE, IL-10 levels were negatively correlated with the acute phase EDSS score (P < 0.05). Conclusion The AS damage exists in the NMO and the damage severity may correlate with patient function defect. AQP4-Ab, GFAP, IL-6 may play important roles in the onset of NMO and the disease aggravating. The decrease of the ApoE and IL-10 may exacerbate NMO damage.

Aim To construct human phage single-chain antibody library associated with esophageal cancer and to screen the specific scFv against Eca109 cells from the liberary. Methods Metastatic periesophageal lymph nodes of esophageal cancer patients were used as the B cells source, the total RNA of these B cells was extracted and prepared as the template of RT-PCR. First, we screened graticulely two pairs of primers of the heavy and light regions separately, then the V_H and V_L fragments were first amplified from the cDNA by the polymerase chain reaction (PCR). Second, the V_H-linker and V_L-linker were amplified from the V_H and V_L fragments. Last, the V_H-linker and V_L-linker were assembled into scFv gene fragments by SOE-PCR,and then Sfi I and Not I restriction site were inlet in it. ScFv gene was cloned into the pCANTAB-5E phagemid. Phagemids were introduced into E.coli TG1 by electrotransformation, followed by rescue of antibody-expressing phage using M13K07 helper-phage superinfection. Recombinant scFv phage library was constracted and PCR was used to identify the insert ratio of scFv antibodies library. Results of SfiI/Not I double digestion reaction positive insert clone were identified by 1.5% agarose gel electrophoresis. The phage library was panned with NHEEC and Eca109 cancer cells in suspension for four rounds. Strongly positive recombinant phage clones were used to infect E.coli HB2151. Expression of soluble scFv was induced by IPTG. Soluble scFv from periplasm were purified by affinity chromatography and identified by SDS-PAGE and Western blot. Cell ELISA , immunohistochemical staining and immunocytochemical staining were used to identify the activity of the soluble scFv. Results The result of agarose gel electrophoresis showed that total RNA of these B cells had two bands of 28 S and 18 S. The size of V_H fragment is about 450 bp,V_L fragment is about 350 bp and scFv is about 850 bp. The competence is 108 cfu??g-1 pUC18 DNA. Randomly digestive reac-tion showed that the positive insert ratio was 91.7% (22/24). After four rounds of panning, the fourth phage yield is 141 times as much as that of the first one. SDS-PAGE and Western blot showed that the MW of the soluble scFv was about 30 ku and the brand of 30 ku was stained. Immunohistochemical staining showed strong stainning of the tissue of esophageal cancer, but not the liver and gastric cancer tissue. Immunocytochemical staining showed significant staining of the esophageal cancer line Eca109. The result of cell ELISA assay revealed that soluble scFv had highly specific and could combined with Eca109 cells, but not with BGC-823 and NHEEC. Conclusion A human scFv phage display library associated with esophageal cancer has been constructed successfully and the specific scFv antibody against Eca109 has been identified from the liberary.

Objective To discuss the effectiveness of knowledge-attitude-practice(KAP)-based health management on type 2 diabetes in community residents.Methods From June 2010 to December 2011,930 type 2 diabetes patients living in Mishi Lane Community were enrolled and assigned to the intervention group(n=494) and the control group(n=436).The KAP mode was used for the intervention group,and the routine management mode was applied to the control group.Fasting blood glucose(FBG),PBG2 h,HbAlc,blood pressure(BP),triglyceride(TG) and body mass index(BMI) were tested and recorded.Student's t test or Chi-square test was used for data analysis.Results Age(t=0.124,P＞0.05),gender(x2=2.0,P＞0.05) and other demographics showed no statistically significant difference between the two groups(all P＞0.05).After one-year KAP-based health management,HbA 1 c of the intervention group reduced to(6.1 ± 1.2)％,FBG and PBG2 h reduced by 1.7 and 3.23 mmol/L,respectively(t=4.926,P＜0.05; t=4.306,P＜0.05; t=4.523,P＜ 0.05).Diet,physical exercises,medication,regular inspection,unhealthy habits and questionare test awaveness of the intervention group were improved(86.03％,82.19％,85.63％,70.45％ 76.11％ and 88.46％,respectively.Drinking,dietary structure,overweight and physical exercises of the intervention group were also improved(14.17％,15.79％,70.65％ and 68.83％,respectively; all P＜0.001).Conclusion The KAPbased health management is proved to be effective in the control of community diabetic patients.

Objective To discuss the clinical value and nursing of the three-endoscope in the treatment of eholedoeholithiasis. Methods 45 eases of choledocholithiasis patients who were treated with LCDE (three-endoscope) were named as the research group.56 patients who received traditional open ab-dominal surgery were set as the control group. The average hospitalization time and satisfaction degree with nursing were compared, t test and χ2 test were adopted. Results The average hospitalization time was shorter and satisfaction degree with nursing was higher in the research group than those in the control group. Conclusions The treatment of choledochohthiasis with three-endoscope is safe and feasible, es-pecially when combined with antibiotics lavage and stone dissolution through naso-biliary duct.The opera-tion can widen the surgical indication,reduce the risk of surgery with little damage,clear stones completely, reduce postoperative complicatioas,make patients recover faster, shorten the hospital stay and achieve the same or better treatment results when compared to the traditional open abdominal surgery.

To investigate the clinical improvement of limping gait in patients with ankylotic hips and limb length discrepancy.Methods:From 1996 to 2005,12 patients with ankylotic hips and limb length discrepancy were treated by distraction osteogenesis with a mono-lateral external fixator and an intramedullary nail.The limb length discrepancy was 6.20-12.50 (median 8.45) cm.Limping gait was classified according to the recommendations of the American Academy of Orthopedic Surgeons/Hip Society and scored according to Harris:no limping scored 11 points,mild limping scored 8 points,moderate limping scored 5 points,while severe limping scored 0 points.Limping gait was severe in all patients pre-operatively and the total score was 0.Results:All patients were followed up for 30.00-46.00 (median 38.55) months,and all reported improvement in limping gait.The gain in length was 6.00-12.50 (median 8.20) cm,and the mean residual limb length discrepancy was 0-0.50 (median 0.20) cm.The total treatment time was 41.00-82.00 (median 61.50) weeks,the lengthening time was 14.00-38.00 (median 29.55) weeks.At the last follow-up,10patients had mild limping gait and 2 had moderate limping gait; the total score was 90.00.The median score was 7.50 (P25 was 8.00,P75 was 8.00).According to Wilcoxon signed rank test,the post-operative limping gait scores were significantly higher than pre-operative (P=0.001 ).Conclusion:Femoral lengthening can improve the limping gait significantly in ankylotic hips and limb length discrepancy.