Objective To construct anti-IL-4R murine anti-human single-chain variable fragment (scFvs) antibodies through BL21 (DE3) prokaryotic expression system. Methods The anti-IL-4R scFv sequence was optimizated on the basis of previous findings. The optimized scFv sequence was analyzed. The recombinant plasmid pET-32a-scFv was constructed. The recombinant plasmid was detected through enzyme identification, and was turned into BL21 (DE3) prokaryotic expression bacteria to express the pET-32a-scFv recombinant protein in E.coli BL21 (DE3). The purification and renaturation were researched, and SDS-PAGE analysis was studied. The molecular weight of ScFv against IL-4R was analyzed by SDS-PAGE. The expression of the fusion protein was detected by Western-blot assay. Results The length of fusion gene scFv-MLT sequence was 761 bp. The molecular weight of the recombinant expression of proteins of anti-IL-4R single antibody was approximately 45 ku. The recombinant proteins showed high specificity with anti-6 × His-tag antibody. Conclusion This experiment successfully constructs pET-32a-scFv prokaryotic expression system of recombinant protein with high immune reactivity, which provides the basis for further study of anti-IL-4R single chain antibody as drug target.

Objective To explore the influence factors of poor myocardial perfusion in patients with ST-segment elevation myocardial infarction( STEMI) after primary percutaneous coronary intervention(PCI). Methods One hundred and forty-three patients with first STEMI who were on admission from April 2010 to May 2014 and underwent primary PCI within 12 hours were enrolled as our subjects. According to the sum-ST-segment resolution(sumSTR)and TIMI myocardial perfusion grade(TMP)after primary PCI,all patients were divided into well myocardial perfusion group( sumSTR ≥ 50% or TMP 2 - 3 grade)and poor myocardial perfusion group(sumSTR < 50% and TMP 0 - 1 grade). The influence factors between two groups were collected and analyzed,including sex,age,pain to balloon time,blood pressure on admission,left ventricular ejection fraction,leucocyte count,neutrophil ratio(NR),high-sensitivity C-reactive protein(hs-CRP),blood lipid,and the history of hypertension,diabetes mellitus. Results The leucocytes count,NR,hs-CRP in patients of poor myocardial perfusion group were(11. 60 ± 3. 57)× 109 / L,0. 84 ± 0. 06 and 9. 80 ± 11. 37 mg/ L,higher than those in well myocardial perfusion group((9. 51 ± 2. 59)× 109 / L,0. 77 ± 0. 11 and(3. 83 ± 5. 58)mg/ L),and the differences were significant(t = 3. 497,P = 0. 001;t = 3. 390,P = 0. 001;t = 3. 973,P < 0. 001). Multiple linear regression analysis showed that neutrophil ratio was independent risk factor of sumSTR in STEMI patient after primary PCI(P = 0. 000). Conclusion The increase of leucocyte count,NR and hs-CRP are related to the poor myocardial perfusion after primary PCI. The increase of neutrophil ratio is an independent risk factor of poor myocardial perfusion.

OBJECTIVETo recognize the characteristics of desmoplastic small round cell tumor (DSRCT) and improve the standard of diagnosis.

METHODSWe retrospectively reviewed the clinical data on the treatment of 2 patients with DSRCT in terms of their conditions, tissue sources, pathologic characteristics, immunohistochemical methods, clinical manifestation, diagnosis, treatment and prognosis.

RESULTSClinical manifestations were complicated. The 2 patients were mis diagnosed before operation. Their tumors consisted of irregular nests of small and round cells, with nuclear hyperchromatism and scant cytoplasm embedded in a plenty of fibrous connective tissues. The edge of the nest was clear, with different sizes and shapes. Immunohistochemically, the 2 patients were positive for CK or EMA, NSE, des and vim of the epithelium, nerve, muscle and interstitial. They died 9 months after operation.

CONCLUSIONSThe tumor may occur in the abdomen, pelvic cavity and other sites, with different clinical manifestations. Routine examination should be replaced by immunohistochemical test for correct diagnosis of the tumor. Prognosis of most patients is not good.

Adolescent ; Carcinoma, Small Cell ; diagnosis ; Female ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Neoplasms, Connective Tissue ; diagnosis ; Retrospective Studies

OBJECTIVE：To study the therapeutic effect and safety of domestic tolterodine tartrate in treating patients with urinary bladder overactivity.METHODS：56 cases of bladder overactivity were divided into two groups randomly：tolterodine and control(oxybutynin)group.The course of treatment was 6 weeks.RESULTS：The effect of tolterodine in treatment group was comparable to that of oxybutynin in control group,however,the adverse reactions in oxybutynin group were more common than those in tolterodine group.CONCLUSION：Tolterodine is a suitable drug to treat bladder overactivity.

The application principle and procedure of reverse engineering and rapid prototyping techniques are introduced in this paper. The description is concentrated on the applications of the above techniques in medical field, especially in reconstructive surgery of mandibular bone defect.
Biomedical Engineering ; Bone Substitutes ; Computer Simulation ; Computer-Aided Design ; Humans ; Mandible ; surgery ; Mandibular Prosthesis Implantation ; methods ; Reconstructive Surgical Procedures ; methods

Objective To investigate the pathological changes and mechanism in the urethra by parturition-induced stress urinary incontinence.Methods Sprague Dawley female rats underwent vaginal balloon dilation for 4 hours immediately after delivery.One week later,the rats were anesthetized and both ovaries were excised.Then a rat model of stress urinary incontinence (SUI) was successfully established.One month after ovariectomy,conscious cystometry and Leak-Point Pressure (LPP) were measured by MP150.Histological examination and Western blotting were performed after functional assays.Results (1) 85％ of rats presented astress urinary incontinence in the model group.(2) The urethras in SUI rats had decreased muscles,and striated muscles showed fragmentized and disorganized.(3) Elastic fibers were long,well organized and tightly connected to the muscle bundles in sham group,while elastic fibers showed fragmentation and disorganization in the model group.(4) The protein expression of vascular endothelial growth factor (VEGF) and blood vessels were reduced in SUI rats as compared with the sham rats.Conclusions Muscle and elastic fibers in the urethra are disrupted in SUI rat.VEGF may play an important role in regulation of pathological changes in urethra.

Objective To explore the role of viral infection in the development of drug eruption in patients with HIV infection, and to evaluate the efficacy of antiviral treatment. Methods This study enrolled 87 HIV-positive patients, including 11 with and 76 without drug eruption, all of whom received highly active antiretroviral therapy(HAART). Clinical data on, baseline CD4+ and CD8+ T cell counts and CD4/CD8 ratio in these subjects were retrospectively analyzed. Results The severity of drug eruption was mild in the 11 HIV-positive patients, with a mean latency period of (14.00 ± 8.10)(range, 8 - 34)days. Of the 11 patients with drug eruption, 7 had liver function impairment, which was not in accordance with the severity of skin lesions. Drug eruption was controlled in all the 11 patients after anti-anaphylactic treatment without withdrawal of antiviral drugs. Compared with 75 HIV-positive patients without drug eruption, the 11 HIV-positive patients with drug eruption showed significantly increased baseline CD4 + T cell counts (493.00 ± 245.68 (range, 42 - 810)/μl vs. 347.81 ± 167.00 (range, 11 - 814)/μl, t = 647.50, P < 0.05), but decreased proportion of patients with baseline CD4+ T cell counts below the lower limit of normal(3/11 vs. 48/75(64.00%), X2 = 3.95, P < 0.05). There were no significant differences between 10 patients with drug eruption and 69 patients without drug eruption in the baseline CD8+ T cell count(1472.30 ± 858.55/μl vs. 1356.59 ± 684.06/μl, P > 0.05), CD4/CD8 ratio(0.40 ± 0.27 vs. 0.29 ± 0.16, P > 0.05), or percentage of patients with a CD4/CD8 ratio below the lower limit of normal (9/10 vs. 68/69 (98.55%), P >0.05). Conclusions The latency period of drug eruption seems to be long in HIV-positive patients receiving HAART, and mild drug eruption can be complicated by liver function impairment in the patients. Relatively high CD4 + counts may be a risk factor for the development and aggravation of drug eruption in HIV-positive patients.

Objective To investigate the distribution characteristics and risk factors of infectious pathogens after open limb fracture surgery.Methods From January 2016 to December 2018,180 patients with open limb fracture admitted to Hangzhou Hospital of Zhejiang Medical and Health Group were selected to observe the infection after operation.Pathogens were isolated and identified by automatic biological analyzer and bacterial identification system,and drug resistance test was carried out by K-B method.Multivariate logistic regression analysis was used to analyze the risk factors of infection after open limb fracture surgery.Results Among 180 cases of open limb fracture,29 cases had postoperative infection,the infection rate was 16.11％.Among 29 cases of post-operative infection,34 strains of pathogenic bacteria were isolated,including 19 strains of Gram-positive bacteria,13 strains of Gram-negative bacteria and 2 strains of fungi.The resistance rate of Staphylococcus aureus to penicillin G was high,and the resistance rate was 80.00％.Univariate analysis showed that there were no statistically significant differences in gender,BMI,injury site,smoking history,hypertension and intraoperative bleeding between the two groups (x2 =0.252,0.416,0.734,0.856,0.572,all P ＞ 0.05).There were statistically significant differences in age,diabetes mellitus,operation time,hospitalization time,wound contamination and drainage tube placement time between the two groups (x2 =21.537,9.664,17.244,15.459,24.327,19.804,all P ＜ 0.05).The single factor analysis showed that age ＞ 60 years old,diabetes mellitus,operation time ＞ 3h,hospitalization time ＞ 14d,wound contamination and drainage tube placement time ＞ 4d were independent risk factors for infection after open limb fracture surgery.Conclusion Postoperative infection rate of open limb fracture is high.Gram-positive bacteria are the main pathogenic bacteria.Postoperative infection is affected by many factors.In order to prevent postoperative infection,specific measures should be taken and antibiotics should be used reasonably in strict accordance with drug sensitivity test.

We examined the biocompatibility and the safety of a-calcium sulfate hemihydrate (CSH)/multi-walled carbon nanotube (MWCNT) composites for bone reconstruction application. The biocompatibility of the CSH/MWCNT composites was evaluated by the measures which taking L929 fibroblast cells cultured in the extracted liquid of the composite soaking solution and putting bone marrow stromal cells planted on the composite pellets in vitro, respectively. The cell proliferation was evaluated by MTT test and further observed using an inverted optical microscope and a scanning electric microscope. The toxicity of the composites was evaluated by acute and subacute systemic toxicity test. Long-term muscle and bone implantation in vivo tests were also conducted. L929 fibroblast cells grew well in the extracted liquid, as well as bone marrow stromal cells that could adhere on the surface of sample pellets and proliferated rapidly. MTT test showed that there were no significant differences between the experimental and control groups (P > 0.05). In vivo test manifested that the composites were no toxicity, no irritation to skin and good for bone defect reconstruction. It was proved that a-calcium sulfate hemihydrate (CSH)/multi-walled carbon nanotube (MWCNT) composites exhibited excellent biocompatibility for the potential application in bone tissue engineering.
Animals ; Biocompatible Materials ; chemistry ; Bone Marrow Cells ; cytology ; Bone Substitutes ; chemical synthesis ; chemistry ; Calcium Sulfate ; chemistry ; Cell Line ; Cell Proliferation ; Fibroblasts ; cytology ; Materials Testing ; Nanotubes, Carbon ; chemistry ; Rabbits ; Stromal Cells ; cytology ; Tissue Engineering ; methods ; Toxicity Tests

OBJECTIVETo study the prevalence of cerebral palsy (here in after referred to CP) in children aged 1 - 6 in Guangxi, China, and its epidemiologic characteristics and relevant risk factors.

METHODSInvestigations on the prevalence and etiology of CP in children at ages of 1 - 6 were conducted in Nanning, Hengxian and Qinzhou of Guangxi from June to December in 1998, with a cluster sampling.

RESULTSTotally, 150 806 children aged 1 - 6 were investigated in Nanning, Hengxian and Qinzhou of Guangxi, with 89 418 boys and 61 388 girls. Among them, 193 children were diagnosed as CP, with a prevalence rate of 1.28 per thousand, higher in boys (136 cases, 1.52 per thousand ) than in girls (57 cases, 0.93 per thousand ), with statistical significance (chi(2) = 9.536 7, P = 0.02). Logistic regression analysis showed that risk factors for CP were children who could not cry after birth (neonatal asphyxia), Apgar score less than eight, delivery at lower level of maternity hospital, number of mother's gravidity, no prenatal checks for mothers, and taking antipyretics and preserved food with salt during pregnancy.

CONCLUSIONSPrevalence of CP in children of Guangxi was at a lower level in comparison with that in other areas at home and abroad, which, maybe, was related with the lower neonatal survival and higher infant mortality.

Cerebral Palsy ; classification ; epidemiology ; etiology ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Infant ; Logistic Models ; Male ; Pregnancy ; Pregnancy Trimesters ; Prevalence ; Risk Factors