Objective To detect the IgM antibodies and epidemiology of pathogens of respiratory tract infection in children ,and to provide the basis for clinical diagnosis and treatment .Methods The serum of 14 379 outpatient and inpatient cases in Tianjin Children′s Hospital from March 2015 to February 2016 were detected by indirect immunofluorescence ,and the respiratory tract in‐fections of different gender ,season ,age and pathogens were analyzed .Results Totally 3 392 specimens (23 .59% ) were positive for IgM antibody detection and the positive rate of MP with 18 .77% was highest ;361 cases were mixed infection ,mainly including two kinds of infection pathogens .The positive rate of respiratory pathogens in male with 21 .46% was significantly lower than in female patients with 33 .92% (χ2 = 274 .73 ,P < 0 .05) .The positive rate in different ages groups (0 - 30 d ,- 6 months ,- 1 years ,- 3 years ,- 9 years ,- 18 years) were 0 .2% ,1 .8% ,15 .36% ,34 .46% ,39 .73% and 30 .73% respectively ,the highest infection rate was found between 3 to 9 years old children ,and the differences among groups were statistically significant (χ2 = 1 407 .87 ,P<0 .05) .The highest rate of MP were found in autumn (24 .42% ) and winter (23 .01% ) ,the highest rate of Influenza B virus (IFu B) was found in spring (15 .13% ) ,the positive rate of Legionella pneumophila (LP) was high in summer (0 .78% ) and autumn (0 .80% ) .Comparison with the 15 departments ,the children of otolaryngology with the positive rate of 43 .9% was the highest . Conclusion The infection ratios of respiratory pathogens were related to gender ,season ,age and pathogen .These findings provided an important reference for clinical diagnosis and treatment in different seasons and population .

Objective To compare the feasibility and efficacy between dexmedetomidine and midazolam in herniorrhaphy surgery for older patients. Methods Sixty American Society of Anesthesiology (ASA) gradeⅠ~Ⅱpatients, treated by herniorrhaphy surgery under local anesthesia,were randomly divided into dexmedetomidine group (n=30) and midazolam group (n=30) .Patients in dexmedetomidine group were given dexmedetomidine at a loading dose of 1μg/kg for 10 min,then they were injected continuously by 0.4μg/(kg·h),whereas midazolam group were given midazolam at a loading dose of 0.06 mg/kg, then 0.04 mg/(kg·h) injected continuously.The mean blood pressure (MAP) and heart rate (HR) were recorded before infusion (T0),incision of skin (T1),15min (T2) and 30 min (T3) after administration and when sutured skin (T4), adverse reaction were also assessed. Results The difference of sedation level was not significant between the two groups (P>0.05) .Compared with T0 , the decrease of HR was significantly more in dexmedetomidine group from T1 to T4 (P<0.05) . Compared with midazolam group , the decrease of HR was significantly more in dexmedetomidine group from T1 to T4 (P<0.05) . Compared with dexmedetomidine group, the rate of respiratory depression and restlessness were more in midazolam group, but bradycardia was lower (P<0.05) .Conclusions Dexmedetomidine is a comparable alternative to midazolam for sedation in herniorrhaphy surgery under local anesthesia. It is associated with better respiration and lower restlessness but with a high incidence of bradycardia.

OBJECTIVETo explore the association between C825T polymorphism of G protein beta3 subunit (GNB3) gene and different Hilit types of essential hypertension (EH) in the Uygur nationality of Xinjiang.

METHODSAccording to Uygur medical theories, EH patients (as the EH group) and non-EH patients (as the control group) were assigned to four Hilit groups. The C825T polymorphism of GNB3 was detected in 161 EH patients and 379 non-EH subjects of different Hilit types by using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) to explore the difference of the genotypes and allelic frequencies and hypertension.

RESULTS(1) In Xinjiang Uygur population, the distribution frequencies of GNB3 C825T polymorphism were in accordance with Hardy-Weinberg (chi2 = 0.871, P = 0.647). (2) There was no statistical difference in the distribution frequencies of three genotypes and two alleles of GNB3 between the EH group and the control group (P > 0.05). (3) There was statistical difference in distribution frequencies of three genotypes between the abnormal Sapra and non-abnormal Sapra group (the sum of abnormal Sewda, abnormal Kan, and abnormal Balhem) (chi2 = 6.905, P = 0.032), especially between the abnormal Sapra and abnormal Balhem groups (chi2 = 10.404, P = 0.006), but there was no statistical difference in distribution frequencies of alleles between the two groups (P > 0.05). (4) In 161 EH patients, there was statistical difference in the distribution frequencies of three genotypes and two alleles between the abnormal Sapra and non-abnormal Sapra group (chi2 = 9.034, P = 0.011; chi2 = 4.701, P = 0.03).

CONCLUSIONSBoth TT genotype and T allele of GNB3 C825T polymorphism might not be associated with EH patients in Xinjiang Uygur populations. However, they were correlated with hypertension patients of non-abnormal Sapra, indicating the pathogeneses of EH with different Hilit types might be different.

Adult ; Aged ; Alleles ; Case-Control Studies ; Essential Hypertension ; Female ; Gene Frequency ; Genotype ; Heterotrimeric GTP-Binding Proteins ; genetics ; Humans ; Hypertension ; classification ; diagnosis ; genetics ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Minority Groups ; Polymorphism, Genetic

Objective:To express rationally engineered antibodies against EGFR and assess their affinity to EGFR and anti-tumor cell migration effect. Methods:L and V_H genes of humanized antibodies against EGFR were designed and synthesized. Genes encoding V_H and C_H were connected and then cloned into a pIRES based bicistronic expression vector. Gene encoding the corresponding L gene was also cloned into the same vector. 293T cells were transfected with the recombinant plasmid and the antibody expression was confirmed by Western blotting. The antibodies were purified by protein A based affinity chromatography. Binding of the humanized antibody to the EGFR was assessed by Surface Plasmon Renainance with Biacore3000, and the biological activity of the humanized antibody was determined by tumor cell invasion test.Results:Three expression vectors were constructed and the humanized anti-EGFR antibodies were expressed and purified successfully. In reducing SDS-PAGE, the antibodies exhibited two bands of approximately 25×10~3and 50×10~3, respectively. Western blot assay showed that the humanized antibodies had recognition specificity to goat-human IgG antiserum. Biacore assay revealed that the humanized antibody C3 binds to EGFR with high affinity(6.13×10~(-10)M). Cell migration test showed that C2,C3 and C5 could suppress growth and migration of tumor cells.Conclusion:Three anti-EGFR humanized antibodies (C2,C3 and C5) have been constructed and expressed successfully, and the C3 antibody retained high affinity for EGFR and showed improved inhibitory effect on tumor cell growth and migration.

OBJECTIVETo observe and compare the effects and adverse events of the three phosphodiesterase type 5 (PDE-5) inhibitors sildenafil, vardenafil and tadalafil used in the same period for the treatment of erectile dysfunction (ED).

METHODSA total of 311 ED patients were treated with the three PDE-5 inhibitors in the same period, 134 with sildenafil, 109 with vardenafil and 88 with tadalafil, and followed up for 2-11 months. Their effects were compared.

RESULTSSildenafil produced satisfactory effect in 72 patients (79.12%), vardenafil in 63 (81.81%) and tadalafil in 52 (78.78%), respectively. And the efficacy rates of sildenafil, vardenafil and tadalafil in the treatment of ED with premature ejaculation were 55.56%, 44.44% and 42.3%, respectively. Our study also showed that the young and the newly married preferred vardenafil, the middle-aged chose tadalafil and the older and long-term users tend to take sildenafil. The main reasons for discontinuing the use of the three inhibitors were their high price, incomplete effect and the patients' worry about their adverse action.

CONCLUSIONThe three PDE-5 inhibitors can produce basically similar effects on ED, all with safety and few mild adverse events.

Adult ; Erectile Dysfunction ; drug therapy ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Phosphodiesterase 5 Inhibitors ; Phosphodiesterase Inhibitors ; administration & dosage

The recombinaut porcine insulin precursor(PIP)produced by Pichia pastoris in shake-flask and 501.fermenter was investigated respectively.The results indicated that 60h induction time length and 2.0%～2.5% methanol addition every day was optimum in shake- flask.The process in 50L fermenter was consisted of batch,feed-batch and induction phases.The relationship between dry cell weight(y) and culture time (t) in growth phase(batch and feed-batch phase)could be described by model y=0.6525e~(0.1907t).Glycerol and ammonia were almost used for cell growth and maintain,and no by-product was observed in batch and fed-batch phase Only 80% ammonia and 70% methanol were used by cell in induction phase.By comparison the results of shake-flask and 50L fermenter,it was concluded that the limit- ing factor in the fermentation of shake-flask and 50L fermenter was dissolved oxygen(DO)and.carbon source,respectively.When scaling the result of shake-flask to 501.fermenter,the control strategy was adapted for 50L fermenter by increasing the feed rate of methanol and the maximum PIP concentration reached 1.72 g/L.

Objective To compaire the physical characteristics of semen morphology before and after semen coagulation-liquefaction under microscope and understand the significance of these characteristics for the determination of semen liquefaction.Methods Semens were placed in sperm counting chamber immediately after collection.The morphological changes of the seminal gelatinous substance during liquefaction were observed under microscope and analyzed.Results Morphological changes during semen liquefaction can be clearly observed.Among 32 samples of semen,forward sperm progression and motility were (30.3?12.4)% and (45.0?14.9)%,(44.9?12.7)% and (59.0?15.3)%,before and after the liquefaction,respectively.The differences were statistically significant (t=-1.130,5.023,P0.05).Conclusions Microscopic observation of the seminal gelatinous substance is a simple and feasible method for objective measurement of sperm liquefaction.These findings suggest it may provide pathological significance on semen liquefaction abnormality in infertility clinic.

Immobilized penicillin acylase was used for bioconversion of penicillin G into 6-APA in aqueous two-phase systems consisted of a light-sensitive polymer PNBC and a pH-sensitive polymer PADB.Partition coefficients of 6-APA was found to be:about 5.78,in the presence of 1% NaCl.Enzyme kinetic showed that reaction reached equilibrium at 7h or so.The 6-APA mole yields were 85.3%(pH 7.8,and 20 ℃) and this value was about 20%higher than control in reaction of single aqueous phase buffer.Partition coefficient of penicillin G(Na) washardly changeable,while partition coefficient of product,6-APA and phenylacetate acid was significantly changeable.Reason is due to Donnan effect of phase systems andhydrophobicity of products.The change of partition coefficients of products also affects bioconversion yield of products.In the aqueous two-phase systems,substrate,penicillin G,products 6-APA and phenylacetate acid are biased in top phase,while immobilized penicillin acylase is completely partitioned in bottom.Substrate,penicillin G enters into bottom phase,and it is catalyzed into 6-APA and phenylacetate acid,then the products enter into top phase.Finally,inhibition of substrate and products is removed to result in improvement of products yield.Moreover,immobilized enzymehashigher efficiency than immobilized cells and occupy smaller volume.Comparing with free enzyme,immobilized enzymehashigher stability,longer use life,completely partitioned in bottom phase and recycle.Bioconversion in two-phase systems using immobilized penicillin acylase showed outstanding advantage.The light-sensitive copolymer forming aqueous two-phase systems could be recovered by laser radiation at 488 nm or filtrated 450 nm light,while pH-sensitive polymer PADB could be recovered by isoelectric point(pH 4.1).The recovery of the two copolymers was 95%~99%.

In this assay, the reaction kinetics, optimum temperature, pH and various influential factors of ATP microbial rapid detection reagent by bioluminescence were studied. The results showed that it's enough for detection system to have 40 ~ 50?g/mL D-Luciferin. The light production decreased fastest in the first minute of reaction, then began to decay slowly. The optimal reaction temperature was 24℃~25℃and the optimal pH was pH 7.2 -7.4 in the reaction system. In addition, when stored at 4℃for 45h, the dissolved reagent solution could keep its 86% activity. When preserved at 25℃, the enzyme activity decreased less for 1h, and degraded gradually as time went by and only left 53. 5% of its activity after 6. 5h. While stored at 33℃, the enzyme activity decreased quickly with the time and only left 59. 1% after 1. 5h. The result indicated that storage temperature was a very important influential factor to the activity of reagent Meanwhile, different chemical substance such as acid, alkali, salt and surfactants inhibited the ATP bioluminescent reaction. When the concentration of NaCl reached 1. 5g/L, it could inhibit 52. 5% light production. Triton X-100, acid, and alkali also had some effects on the reaction, while CTAB, SDS and TCA would inhibit the bioluminescent reaction seriously.

Background Chronic administration with inhibitor of nitric oxide synthase (N_?-nitro-L-arginine methyl ester,L-NAME) induces persistent hypertension,cardiovascular remodeling,surrounding vascular fibrosis, necrosis and hypertrophy of myocardium,and inflammation in cardiovascular system.Local RAS involves in hyper- tension and remodeling of cardiovascular system,via increasing production of oxygen free radicals (OFR).Objec- tive To elucidate the preventive and therapeutic effect of antioxidant Ebselen and/or VitE on hypertensive heart damage in NO~- deficient rats induced by L-NAME.Methods Thirty-two Wistar rats were administered with L- NAME 50 mg/(kg?d) by gavage for 8 weeks,and randomized to received a placebo(L),or Ebs(S,30 mg/kg?d), or VitE(V,40 mg/kg?d) or Ebs 30 mg/(kg?d)+VitE 40 mg/(kg?d),with 8 normal Wistar as control. Body mass and SBP were measured fortnightly.Plasma and homogenate of heart were collected for NO,Ang Ⅱ, GSH-PX,MDA and O_2~- determination.Results Eight weeks after L-NAME administration,SBP in experimental groups was obviously higher than that of control (P