Objective To construct cDNA library of alveolar macrophages after lipopolysaccharide (LPS) and dexamethasone (Dex) treatment for exploring the protein molecule interactive with glucocorticoid receptor (GR) in condition of inflammation. Methods After the cultured AMs were treated with LPS (10mg/ml) and Dex (10 -5 mol/L) for 4h, total RNA was extracted from AMs, then the cDNA was synthesized from total RNA of AMs and amplified using primers SMARTⅢ TM and CDSⅢoligo (dT) as the base of recombination. The purified PCR products as well as the linearized plasmid pGADT7-Rec were co-transformed into the competent yeast AH109. They were recombined by yeast homologous recombinase in the yeast cells and became the active cyclic plasmid. The transformed yeasts grew in the SD/-Leu plates. All the growing clones were harvested and then constituted the cDNA library. Furthermore, the bait pGBKT7-rGR transformed the yeast AH109 of library was constructed, and the protein molecule interactive with GR was screened. Results cDNA library of AMs was constructed with high multiplication and good capacity. 1.07?106 recombinants were obtained from the cDNA library. The amplified PCR fragments were between 0.3-1.5kb in size. One true positive clone, obtained by screening the cDNA library, was confirmed to be BAG-1 by sequencing and BLAST. Conclusion The yeast two-hybrid cDNA library of AMs was successfully constructed by Clontech SMART method; GR can interact with BAG-1 in yeast cells.

90%) and 1 incompletely (

Objective To study the effect of cardiac troponin T(cTnT)on the prognosis of chronic heart failure in the elderly patients.Methods Serum level of cTnT,and serum activities of creatine kinase (CK)and creatine kinase-MB(CK-MB)were measured in 60 elderly patients of chronic heart failure with varied classes of cardiac function.Cardiac events,including malignant cardiac arrhythemia,repeated or aggravated heart failure or cardiac death,in patients with increased cTnT during hospitalization were compared to those without it.Results Serum level averaged(89.1?13.6)mmol/L for CK and(7.9? 1.4)mmol/L for CK-MB in 20 patients with Class Ⅱ cardiac function by New York Heart Association (NYHA)classification and without increased cTnT,(88.2?13.4)mmol/L for CK and(7.5?1.8) mmol/L for CK-MB in 19 patients with Class Ⅲ cardiac function and four of them with increased cTnT,and (93.3?14.2)mmol/L for CK and(8.1?1.6)mmol/L for CK-MB in 21 patients with Class Ⅳ cardiac function and 14 of them with increased cTnT.Malignant cardiac arrhythemia occurred in six,repeated heart failure in eight,and death in three of 18 patients with increased cTnT,and one,two and one in 22 patients without increased cTnT,respectively.Incidence of cardiac events was higher in patients with increased cTnT than those without it(P

AIM:To evaluate the morphological and functional changes of retinas induced by treatment of tunicamycin with optical coherence tomography (OCT) in rats.METHODS:Totally 60 SD rats were randomly divided into 3 groups (20 in each group), 0.5mg/kg (in low dose group), 1.5mg/kg (in high dose group) tunicamycin were injected into vitreous cavity and saline (9g/L NaCl) were injected in the same dose as a control group.Changes of retinas were observed by OCT on the 1,7 and 14d after treatment of tunicamycin.Then the rats were sacrificed, retinas were taken out and embedded by the paraffin, tissue sections and the HE staining were performed.RESULTS:OCT results suggested that tunicamycin played damage effects on retinal morphology and structure which appeared a time-and dose-dependent.Fundus photography results suggested that 2wk after tunicamycin treatments, with the gradually changing of tunicamycin concentration, peripheral retinal and macular region became pale color gradually, edema occurred in optic disk, retinal vessels appeared thinner in the high dose group, optic nerve came out atrophy.Fluorescein angiography confirmed that tunicamycin injection in vitreous cavity 2wk later, retinal vessels injury occurred, resulted in leaking of intravascular contrast agent from peripheral to the central part of the retinas.Electrophysiological data showed that retinal electrogram occurred disorder induced by tunicamycin, such as the amplitude of a wave, b wave decreased gradually, even closed to zero, which was very different from control significantly (P<0.05).HE staining of paraffin sections showed that retina injuries induced by tunicamycin were in dose-time dependent, which was consistent with the results of OCT.CONCLUSION: Clinical retinal diseases could be simulated by retinal damage animal model induced by tunicamycin treatment.OCT detection offered real-time images of the retinal cross-section, which provided a helpful non-invasive method for detecting and evaluating the retinal damages.

Objective To investigate the ultrasound characteristics of prenatal fetus otocephaly . Methods Three prenatal fetus with otocephaly were examined with two -and three-dimensional ultrasoundand examined results were compared with the those of induced labor or autopsy ,and ultrasound characteristicsof prenatal fetus were analyzed and summarized .Results The ultrasound performance of three prenatal fetuswith otocephaly and the examination of the appearance after induced labor showed :(1) The most intuitiveinitial sonographic performance of otocephaly was manifested by the absence of stomach bubble andoverabundance of amniotic fluid.Among three fetus,one fetus had overabundance of amniotic fluid at the midstageof pregnancy,one fetus had normal amniotic fluid at the mid-stage of pregnancy and one fetus hadextremely high amount of amniotic fluid and absence of stomach bubble at late stage of pregnancy .(2) Allthree fetus showed agnathy and synotia (shifts of both ears to the midline) and microstomia deformity.(3) All three fetus had associated complications with deformity in other systems including two cases of patients withcleft lip and palat,both were the fracture unilateral cleft lip derived from small mouth .One fetus withdysmelia and one fetus with complicated cardiovascular deformity and situs inversus and .(4) The results ofexamination after induced labor or autopsy were consistent with those of the prenatal ultrasound examination . Conclusions Prenatal ultrasound examination is an effective and feasible means for the diagnoses ofotocephaly.When the symptoms of “absence of stomach bubble and extremely high amount of amniotic fluid ”occurred,the fetal ear and submaxilla should be examined to confirm stand -alone otocephaly prenatally.

Objective To investigate the aetiology of acute exacerbations of COPD (AECOPD),and the effects of moxifloxacin in the treatment of AECOPD.Methods Patients with stable COPD based on GOLD criteria were included in the study.Sputum collected at first exacerbation was analyzed for bacteria count and culture.IL-6,IL-8 and TNF-?were measured by enzyme- linked immunosorbent assay (ELISA).Eligible patients were randomized to receive moxifloxacin (400 mg qd for 5 days) or ce- faclor (250 mg q8h for 7 days).Efficacy parameters were evaluated at 7 and 14 days after treatment initiation and 1 year later. Results Of the 46 patients with moderate or severe COPD (male 38,moderate 24),21 (45.65%) were microbiologically evalu- able at baseline.The main pathogen was Haemophilus influenzae (10/21).Clinical efficacy rate was 87.0% in moxifloxacin group and 82.6% in cefaclor group.Bacterial eradication rate was 80.0% and 72.7% respectively.The difference between groups was not statistically significant in terms of clinical or microbiological efficacy.In moxifloxacin arm,the frequency of ex- acerbation was 2.6?1.0,significantly lower than control arm (3.5?1.4,P

Objective A distintegrin and metalloproteinase 33 (ADAM33) is one of the asthma susceptibility gene, which is closely related to the pathogenesis of asthma and airway hyperreactivity.The aim of this study was to evaluate the expression of serum ADAM33 in patients with different severity of asthma and the relation to airway chronic inflammation through clinical trials.MethodsPatients diagnosed as mild-to-moderate asthma (n=54), severe asthma (n=35) and healthy controls (n=30) were recruited from May 2015 to May 2016.The serum IgE, ADAM33, IL-4 and IL-13 1evels and Eosinophil (EOS) in peripheral blood were detected, and the correlation analysis was performed Results The serum levels of ADAM33 in mild-to-moderate asthma group, severe asthma group and healthy control group were 23.8±6.21pg/mL,64.8±12.8pg/mL and 18.3±4.49pg/mL, respectively.The ADAM33 levels in mild-to-moderate asthma group and severe asthma group were higher than those in control group (P<0.05).The ADAM33 levels in severe asthma group were higher than those in mild to moderate asthma group.(P<0.05).The correlation analysis showed that ADAM33 had positive correlation with IL-4 and IL-13(r=0.79 and r=0.81), but no correlation with EOS and IgE(r=0.54 and r=0.46).Conclusion The expression of serum ADAM33 was up-regulated in asthmatic patients along with the severity of asthma.ADAM33 was positively correlated with serum IL-4 and IL-13, implying that the expression of ADAM33 may be regulated by Th2 type cytokines.

Objective To investigate the scope of application of precise hepatectomy and its value.Methods The clinical data of 112 consecutive patients with liver neoplasm who received hepatectomy at Renji Hospital from November 2006 to March 2009 were retrospectively analyzed.Precise hepatectomy technique was applied to 88 patients(precise group),whereas pringle maneuver was applied to the rest 24 patients(prince group).Patients in precise group had undergone lobectomy,segmentectomy or local resection,while patients in pringle group received segmenteetomy or local resection.The perioperative conditions of patients in the 2 groups were compared via Fisher exact probability and l test.Results No perioperative mortality,hepatic failure,reoperation due to massive hemorrhage or bile leakage was observed.The blood transfusion rate,blood loss,postoperative total bilirubin(Tbil),prothrombin time(PT)were 7%(2/29),220 ml,20 μmol/L,13 seconds in patients who received segmentectomy and local resection in precise group,and were 4%(1/24),210 ml,19μmol/L and 13 seconds in patients who received segmentectomy and local resection in pringle group,with no significant difference(t=0.248,0.263,0.246,P>0.05).The operation time,postoperative white blood cell (WBC)count.alanine aminotransferase(ALT)value and fever incidence were 60 minutes,7.5×10~9/L,66 U/L,10%(3/29)in patients who received segmentectomy and local resection in precise group,and were 15 minutes,14.0×10~9/L,335 U/L and 42%(10/24)in patients who received segmentectomy and local resection in pringle group,with significant difference(t=4.962,4.961,4.959,P<0.05).In precise group,the blood transfusion rate,postoperative WBC count,ALT value.PT and fever incidence were 9%(4/45),8.3×10~9/L,153 U/L,17 seconds and 13%(6/45)in patients who received Iobectomy,and were 12%(5/43),8.2×10~9/L,133 U/L,14 seconds and 14%(6/43)in patients who received segmentectomy or lncal resection,with no significant difference (t=1.652,1.225,1.236,P>0.05);the blood loss,operation time and postoperative Tbil level were 350 ml,250 minutes and 32μmol/L in patients who received lobectomy.and were 240 ml,150 minutes and 21 μmol/L in patients who received segmentectomy or local resection(t=4.915,4.967,4.829,P＜0.05).Conclusions Precise hepatectomy can decrease damage to patients,but it should be applied in selected patients according to the tumor location and the excision range.Precise hepatectomy is recommended to be applied in lobeetomy.

OBJECTIVETo search for the optimal strategies for sperm collection from the patient with temporary penile erectile dysfunction (ED) on the day of oocyte pick-up ( OPU) in in vitro fertilization embryo transfer (IVF-ET).

METHODSWe retrospectively analyzed 93 cases of temporary ED on the OPU day of IVF-ET from January 2011 to May 2014, with fresh semen for 45 cases (group A), cryopreserved sperm before oocyte retrieval for 30 cases (group B), and frozen oocytes for 18 cases (group C). Group A was again subdivided into A1 (n = 18) and A2 n = 27) , the former intervened with oral sildenafil while the latter left untreated. We compared the rates of fertilization, high-quality embryo, and pregnancy among different groups.

RESULTSNo statistically significant differences were found among groups A, B and C in the age of the males and females, duration of infertility, numbers of obtained and mature oocytes, and rates of cleavage, or in the percentages of normal fertilization (80.78% vs 80.43% vs 84.77%), high-quality embryo (53.27% vs 52.97% vs 47.69%) and pregnancy (60.00% vs 56.77% vs 44.44%) (all P > 0.05). The rate of 3PN was markedly lower in group C (0.63%) than in A (9. 61%) and B (4.34%) (P < 0.05). There were no significant differences between groups A1 and A2 in the age of the males and females, duration of infertility, numbers of obtained and mature oocytes, and the rates of fertilization, cleavage, high-quality embryo, and pregnancy (all P > 0.05).

CONCLUSIONOn the OPU day of IVF-ET, oral sildenafil can help temporary ED men to achieve penile erection and ejaculation without affecting the outcomes of assisted reproduction. Cryopreserved sperm can be used in case of predicted temporary ED and frozen oocytes can also be employed if sperm retrieval fails. However, to avoid puncture injury to the epididymis or testis, fresh semen should be the first choice.

Cryopreservation ; Embryo Transfer ; Erectile Dysfunction ; drug therapy ; physiopathology ; Female ; Fertilization ; Fertilization in Vitro ; Humans ; Male ; Oocyte Retrieval ; Oocytes ; Penis ; Pregnancy ; Retrospective Studies ; Sildenafil Citrate ; therapeutic use ; Sperm Retrieval ; Spermatozoa

Objective:To explore the influence of tumor cells on CD4+CD25+Treg cells via TLRs.Methods:The numbers changes of CD4+CD25+Treg cells in the co-culture system of Lewis lung cancer cells and splenic lymphocytes were detected by flow cytometry;The expression of Foxp3 and TLR1-9 mRNA after co-culture were detected by RT-PCR;TLR9 expression on Lewis lung cancer cells was blocked by TLR9 receptor antagonist chloroquine.Results:Compared with control group,the number of CD4+CD25+Treg cells and Foxp3 mRNA expression were significantly increased in the co-culture group(P