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 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 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 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 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.

Objective: To set a quantified diagnostic standard for large intestinal cancer of spleen qi deficiency syndrome. Methods: The spleen qi deficiency syndrome was identified by experts on the basis of clinical epidemiological investigation of 311 patients suffering from large intestinal cancer. Corresponding points were assigned to the correlative factors (traditional Chinese medicine symptoms) on the basis of symptom differences between spleen qi deficiency syndrome and non-spleen-qi-deficiency syndrome. The best threshold was determined by receiver operating characteristic curve (ROC) according to syndrome differentiation from expert team, and the quantified diagnostic standard was established. The syndrome identification from the expert team which was regarded as golden standard was tested retrospectively. Results: All the traditional Chinese medicine symptoms possibly related to spleen qi deficiency syndrome were analyzed based on the opinions of experts, and 28 symptoms were confirmed as candidate correlative factors. The occurrence of 11 symptoms between spleen qi deficiency syndrome and non-spleen-qi-deficiency syndrome showed statistical differences by means of crosstabs analysis (P<0.05). The 11 symptoms were filtered by logistic regression analysis, and tiredness, fatigue, loose stool, and poor appetite were finally determined as the symptoms relative to large intestinal cancer. These four symptoms were analyzed with conditional probability conversion and endowed with 16, 11, 4 and 8 points respectively. The diagnostic standard of spleen qi deficiency syndrome of large intestinal cancer was over 13 points. The sensitivity, specificity and accuracy of retrospective examination were all above 80%, and its positive likelihood ratio was 9.89. Conclusion: The quantified diagnostic standard for spleen qi deficiency syndrome of large intestinal cancer is in accordance with clinical characteristics of large intestine cancer and the characteristics of TCM syndrome diagnosis.

Aim To investigate the effect of resveratrol on proliferation and differentiation in K562 cells. Methods K562 cells were treated with different con-centrations of resveratrol for 6d. The colony number of K562 cells was detected with semi-solid culture assay. Expression of GATA-1 and PU. 1 in K562 cells was re-spectively measured with immunocytochemistry and Western blot. Expression of differentiation related anti-gen, CD11b, CD14 and CD42b, was measured with flowcytometry on K562 cells. Results Resveratrol
could significantly decrease the colony number of K562 cells in a dose-dependent manner, and enhance the ex-pression of GATA-1,PU. 1,CD11b, CD14 and CD42b in K562 cells. Conclusion Resveratrol could inhibit the proliferation and induce differentiation of K562 cells via up-regulating the expression of GATA-1 and PU. 1 protein.

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 explore the possible role of vitamin D in the pathogenesis of IgA nephropathy (IgAN). Me-thods After the IgAN model was successfully induced at 12 weeks, the BALB/C mice were randomly divided into IgAN group (n=15) and IgAN+VitD group (n=15). The nephrosis mice were administrated with 100 μl/d propylene glycol or propyl-ene glycol+1,25(OH)2D, 3 ng/(100g?d), for 6 weeks. The control group was setted (n=15). The level of 24 hour urine protein was determined at week 0, 12 and 18. At week 18, the levels of serum 25(OH)D, ifbroblast growth factor 23 (FGF23) and galactose-deifcient IgA1 (Gd-IgA1) were detected. The mRNA and protein expressions of interleukin-21 (IL-21) in Peyer’s patches (PPs) were detected by lfuorescent quantitative reverse transcription-polymerase chain reaction and western blot respectively. The protein expression of Bcl-6 was detected by western blot. The percentages of Tfh cells/T lymphocytes, B220+IgM+/B lympho-cytes, B220+IgA+/B lymphocytes, B220-IgA+/B lymphocytes in PPs were determined by lfow cytometry. Results Compared with control group, the levels of 24 hour urine protein, FGF23 and Gd-IgA1 were increased, serum 25(OH)D was decreased, the mRNA and protein expressions of IL-21 and the protein level of Bcl-6 were increased, the percentages of Tfh cells/T lym-phocytes, B220+IgM+/B lymphocytes, B220+IgA+/B lymphocytes, B220-IgA+/B lymphocytes were elevated in IgAN group (P<0.05). These indicators were improved in IgAN+VitD group. Compared with the IgAN group, the differences were statisti-cally signiifcant (P<0.05), however compared with control group, some indicators showed no signiifcant differences (P>0.05). Conclusions 1,25(OH)2D may protect the microenvironment of PPs in IgAN through inhibiting the differentiation of Tfh cells and B cells and the generation of Gd-IgA1.

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.