The interaction between the antibody and the corresponding target molecule determines the characteristics of immunoassay. In this study, a single chain variable fragment antibody (scFv4C7) derived from the hybridoma strain 4C7 were prepared via genetic engineering technique. The recognition properties of scFv4 C7 was determined and compared to those of the parent monoclonal antibody by indirect competitive enzyme-linked immunosorbent assay(ic-ELISA). Three dimensional structure of the scFv4C7 was presented by Swiss-Model, and sulfathiazole ( STZ) was docked to the scFv4C7 model to obtain the structure of the binding complex. The results from the ic-ELISA showed that the binding properties of scFv4C7 were comparable with the parent monoclonal antibody and STZ was almost completely buried in a deep binding pocket formed by the heavy chain and light chain of the antibody. The interaction between STZ and scFv4C7 was more closely related to the heavy chain and the complementarity-determining region ( CDR ) H3 loop played more important role than other CDR loops. The study preliminary provided the necessary structural information for the preparation of antibody with broader specificity and higher affinity.

Objective To observe the effects of Jinghua Weikang Capsule on NF-κB p65 expression in gastric mucosa of Helicobacter pylori (Hp) infected mice;To discuss its mechanism of action. Methods Totally 44 SPF KM mice were randomly divided into blank group, model group, triple therapy group and Jinghua Weikang group. Except for the blank group, Hp infected model was set up by short term immunosuppression combined with peroral infection. The treatment strategies were triple therapy group with lansoprazole, metronidazole and clarithromycin for 1 week, Jinghua Weikang group with Jinghua Weikang volatile oil for 4 weeks, model group with sterile water for 4 weeks, and blank group free of any treatments. All the subjects were sacrificed in the 4th week after the treatment. NF-κB p65 was detected through real-time PCR and immunohistochemical method. Results The relative quantitative expressions of NF-κB p65 mRNA were 3.27±0.53 in blank group, 2.18±0.50 in control group, 4.19±1.21 in triple therapy group, and 7.70±1.41 in Jinghua Weikang group (χ2=86.303, P<0.001). Compared with model group, the expressions of NF-κB p65 mRNA of other three groups increased statistically differently (P=0.004), and there was also statistical difference between triple therapy and Jinghua Weikang group. Through immunohistochemical tests, the values of NF-κB p65 mean optical density of nucleus were 25.18± 7.42 in blank group, 31.17±2.98 in model group, 30.06±4.69 in triple therapy group, and 26.94±5.19 in Jinghua Weikang group (χ2=10.960, P=0.012). The values of NF-κB p65 mean optical density of cytoplasm had no statistical difference (F=1.922, P=0.134), and the nucleus/cytoplasm values were 0.23±0.06 in blank group, 0.28±0.03 in model group, 0.26±0.04 in triple therapy group, and 0.24±0.04 in Jinghua Weikang group (χ2=14.527, P=0.002). The statistical analysis of the nucleus/cytoplasm values showed significance between blank and model group (P=0.002), while triple therapy group was still equal to model group (P=0.420). Jinghua Weikang group was lower than model group (P=0.022) and equal to blank group (P=0.750). Conclusion Jinghua Weikang Capsule can effectively reduce the expression of NF-κB p65 in protein level in Hp infected mice.

Objective To study the treatment of differentiated thyroid carcinoma. Method The clinical data and, in partricular, the treatment modalities of 173 cases of differentiated thyroid carcinoma were retrospectively analyzed.Results All of the 173 patients were treated by resection and subsequent endocrine therapy.Different types of operations were selected in accordance with the clinical stages.No perioperative deaths occurred.There were altogether 30 operative complications including nerve injury.Ninety-one patients were followed up for over 5 years and no deaths occurred.Jugular lymphatic metastases occurred in 4 patients, sternum metastases in 2 and lung metastasis in 1.Conclusion Surgical resection and endocrine therapy are highly effective for the treatment of differentiated thyroid carcinoma.

Objective To study the diagnosis and treatment of hyperthyroidism companied with thyroid carcinoma . Methods The clinical data of 11 cases of hyperthyroidism with thyroid carcinoma were retrospectively analysed. Results 9 of 11 cases were diagnosed preoperatively,and comfirmed by frozen section intra-operatively and underwent suitable operation. In the other two cases the final diagnosis was made by pathological examination postoperatively, and re-operation was performed on 1 case . All the patients were followed up for 1～16 years and neither hyperthyroidism nor thyroid carcinoma recurred. Conclusion It is difficult to make diagnosis of hyperthyroidism with thyroid carcinoma preoperatively. B mode ultrasonography may find some nodes in enlarged thyroid; but fine needle aspisation biopsg(FNA) has high false negative diagnostic rate. Intra-operative frozen section examination is important in the diagnosis of hyperthyroidism with thyroid carcinoma .

Objective To evaluate the effect of combined therapy for inflammatory cancer of breast (ICB). Methods A retrospective analysis was made on the clinical manifestation, pathological type and treatment modalities of 25 patients with ICB from 1985 to 2000.Results All of the 25 patients were treated with mastectomy, including typical Halsted mastectomy in 12 cases, typical Halsted mastectomy with reconstruction of latissimus dorsi flap in 4, and palliative mastectomy in 9. Of the 21 patients 20 received preoperative chemotherapy; and all the 21 patients received postoperative chemotherpyl; while hormonal therapy was used in 11 cases. A mean survival was 23 months (0.5～5years) in 14 followed up patients; and now 5 patieats remain alive (2months～5 years). Conclusions A combined therapy consisting of chemotherapy, surgery and hormonal therapy may improve the prognosis of ICB.

This article summarized the technique research and application status of near-infrared spectroscopy (NIRS), in order to explore hypoxia,qi-deficiency and their connections with typical symptoms. It also discussed NIRS acquired oxygen saturation (SaO2) and its relevant means and methods for information quantification ofqi-deficiency. The accurate obtained SaO2 value was applied in the association with the strength of chest distress in the subjective sensation due toqi-deficiency. According to current data, the normal value,qi-deficiency degree and uncomfortable degree of subjective sensation among patients of different altitudes and ages were summarized. It can be applied to assist TCM physicians in the quick quantification of qi-deficiency.

Objective To observe the effects of deep electroacupuncture on carlilage tissue in knee osteoarthritis (KOA) rabbits. Meth-ods 40 New Zealand rabbits were randomly divided into normal group (A, n=10) and model group (n=30). The model group was modeled KOA with Hulth-Telhag way, and identified with X-ray. Then they were divided into no-treated group (B, n=10), deep electroacupuncture group (C, n=10) and routine electroacupuncture group (D, n=10) randomly. The groups C and D accepted electroacupuncture since 6 weeks after modeling, for 4 weeks. They were measured with pH of joint fluid, observed structure and pathology of cartilage under transmission electron microscope, detected apoptosis index, and determined the expression of acid-sensing ion channel 1 (ASIC1), p38 mitogen-activated protein kinases (p38MAPK) and p53 with Western blotting, and distribution of ASIC1 with immunohistochemistry in cartilage tissue. Re-sults The pHs of joint fluid from high to low were ranged as the groups A=C>D>B (P<0.01). The cartilage structure was more complete in the groups A, C and D than in the group B. The apoptosis rates from less to more were ranged as the groups A=C

Through introducing mutations into ribosomes by obtaining spontaneous drug resistance of microorganisms, ribosome engineering technology is an effective approach to develop mutant strains that overproduce secondary metabolites. In this study, ribosome engineering was used to improve the yield of butenyl-spinosyns produced by Saccharopolyspora pogona by screening streptomycin resistant mutants. The yields of butenyl-spinosyns were then analyzed and compared with the parent strain. Among the mutants, S13 displayed the greatest increase in the yield of butenyl-spinosyns, which was 1.79 fold higher than that in the parent strain. Further analysis of the metabolite profile of S13 by mass spectrometry lead to the discovery of Spinosyn α1, which was absent from the parent strain. DNA sequencing showed that there existed two point mutations in the conserved regions of rpsL gene which encodes ribosomal protein S12 in S13. The mutations occurred a C to A and a C to T transversion mutations occurred at nucleotide pair 314 and 320 respectively, which resulted in the mutations of Proline (105) to Gultamine and Alanine (107) to Valine. It also demonstrated that S13 exhibited genetic stability even after five passages.
Genetic Engineering ; Macrolides ; metabolism ; Point Mutation ; Ribosomal Proteins ; genetics ; Ribosomes ; metabolism ; Saccharopolyspora ; metabolism

Objective To summarize the clinical characteristics and outcome of renal cyst infection in patients with autosomal dominant polycystic kidney disease (ADPKD). Methods Clinical data of 40 ADPKD patients with 43 episodes of renal cyst infection admitted in Shanghai Changzheng Hospital from 1st January 1991 to 31st December 2010 were retrospectively analyzed.Differences of microbiological data and treatments between 1st January 1991 to 31st December 2000 and 1st January 2001 to 31st December 2010 were compared. Results Among 473 identified patients with ADPKD and 662 episodes of hospitalization,40 patients had 43 episodes of renal cyst infection,including 8 definite and 35 likely cases.Microbiological documentation was available for 34 episodes (79.0％),Escherichia coli accounting for 82.4％ of all retrieved bacterial strains.Resistant Escherichia coli to quinolone and certain β-lactamine increased in recent decade.Clinical efficacy of initial antibiotic treatment was noted in 69.8％ of episodes. Antibiotic treatment modification was more frequently required for patients receiving initial monotherapy compared with those receiving combination therapy.In the first ten-year group,initial combination therapy and clinical efficacy were noted in 30.0％ and 60.0％ of episodes respectively,and hospital stay was (20.2±6.7) d.In the second ten-year group,initial combination therapy and clinical efficacy were noted in 61.9％ and 78.2％ of episodes respectively,and hospital stay was (16.3±3.2) d.Large infected cysts (diameter ＞5 cm) frequently required drainage. Conclusions In renal cyst infection,the source of the organisms is often a gram negative enteric organism.Empiric therapy is often initiated with two antibiotics.The drainage of large infected cysts remains the main treatment for cyst infection.

Objective To analyze the causes of 652 hospitalizations in the patients with autosomal dominant polycystic kidney disease (ADPKD).Methods The medical records of all ADPKD inpatients in our hospital from January 1,1990 to December 31,2010 were collected.The differences of hospitalization causes in different age,gender and period were analyzed.Results (1)In 652 hospitalizations,the most common cause was lumbar pain (15.2％),followed by cystic bleeding (14.6％),aggravating renal failure (10.1％),dialysis-related problems (9.4％),renal transplant related issues (8.3％),renal replacement therapy for ESRD (8.0％),urinary tract infection (6.4％),end stage renal failure (5.8％),hypertension (4.1％),renal cyst volume enlargement (3.7％),finding polycystic kidney disease (2.1％),urinary lithiasis (1.8％) and others (10.4％).(2)Younger patients were admitted into hospital because of polycystic kidney bleeding and finding PKD.With the increase of patients age,hospitalization due to dialysis-related problems increased,while many middle-aged patients were hospitalized because of back pain.(3)Male patients were admitted into hospital for aggravating renal failure,ESRD,kidney transplantation-related problems and urinary lithiasis,while female patients mainly for lumbar pain,dialysis-related problems and urinary tract infection.(4)The proportion was significantly reduced with time of finding PKD,renal failure and polycystic kidney bleeding,the proportion of renal cysts increasing and aggravating renal failure increased,there was a significant increase in the proportion of patients with hypertension,while a significant decrease in the proportion of patients with uncontrolled hypertension,and the average SBP was also significantly reduced.Conclusions The highest rate of hospitalization of ADPKD patients is in 40 to 60 age group.Cause of admission varies with age and gender,and changes with the change of time.Over the past decade,the proportion of hospitalization due to renal cysts enlargement and renal failure aggravation increased significantly.The incidence of hypertension is higher than that in the first 10 years,but hypertension control rate increases compared with the previous.Prevention should focus on finding the suppression measures of renal cysts enlargement.