Purpose　To investigate the role of urokinase plasminogen activator (uPA),uPA receptor (uPAR),tissue type plasminogen activator (tPA) and plasminogen activator inhibitor 1 (PAI-1) in the invasiveness of human breast cancer cells.　Methods　Three human breast cancer cell lines with different invasive ability were taken as research targets.RT-PCR and milk plates methods were used to detect the expression of uPA system members and the PA activities,respectively.Modified Boyden's chamber model was employed to detect the invasive ability of cancer cell.　Results　MDA-MB-231 could express high level of uPA,uPAR,PAI-1 and low level of tPA.MDA-MB-435 could express lower level of uPA and hight level of tPA,but no PAI-1 and uPAR were detected.MCF-7 could express lower level of uPAR and high level of PAI-1,but no uPA and tPA were detected.MDA-MB-231 cells showed the highest total PA and uPA activity.MDA-MB-435 cells also showed high total PA activity,but almost all the activity owed to tPA.MCF-7 showed almost no PA activity.Correlated with their PA activities,MDA-MB-231 was found the most invasive in vitro,followed by MDA-MB-435,and MCF-7 almost had no invasive ability.The antibodies against uPA and uPAR were significantly effective in reducing the matrigel invasiveness of MDA-MB-231 by approximately 83.1% and 43.9% respectively (P<0.05).　Conclusions　Co-expression of uPA,uPAR and PAI-1 in human breast cancer highly correlates with the invasiveness in vitro.

Objective To investigate the significance of optimization of bedside Gram staining of sputum smear in the early diagnosis and antimicrobial treatment for ventilator-associated pneumonia(VAP)patients. Methods The data of patients with VAP undergoing mechanical ventilation over 48 hours in the Department of Critical Care Medicine of Tianjin Fourth Central Hospital from June 2009 to June 2014 were analyzed. The patients were divided into two groups according to whether or not bedside Gram staining of sputum smear was used or not. The sputum samples from lower respiratory tract of all VAP patients were collected daily with tracheal catheter. In empirical examination group(from June 2009 to December 2011,n=43),the patients received antibiotics at the time of onset of VAP, selection of antibiotics depended on the information of bacterial epidemiology of the intensive care unit(ICU),and also existence of high risk factors of multi-drug resistant bacteria. In target treatment group(from January 2012 to June 2014,n=43),the patients received antibiotics according to the results of bedside instant sputum smear examination and empirical antibiotic regime. The correlation between the results of sputum smear examination and culture result was analyzed. The levels of body temperature,white blood cell(WBC)count,procalcitonin(PCT)level,and high sensitivity C-reactive protein(hs-CRP)were measured on the 1st day and 3rd day. The length of antibiotics treatment, duration of mechanical ventilation,and the time of ICU stay were recorded for both groups. Results There were 512 qualified sputum specimens for culture,from which 336 pathogens were found,and 358 strains of pathogenic bacteria were found from microscopic examination of 512 qualified sputum smear. The coincidence rate of results of bedside examination of sputum smear and that of sputum culture was 78.32%(401/512). The diagnostic acumen of the former was 85.42%(287/336),specificity was 64.77%(114/176),positive predictive value was 80.17%(287/358),and negative predictive value was 74.03%(114/154). On the 1st day,no statistical differences in infection index between the two groups could be found,but on the 3rd day,the results were significantly improved in both groups. Compared with the empirical treatment group,the body temperature,WBC,PCT and hs-CRP in the target treatment group were significantly lower〔body temperature(℃):36.83±0.69 vs. 37.64±0.71,WBC(×109/L):7.91±2.75 vs. 9.66±3.39,PCT(μg/L):7.14±3.89 vs. 10.14±4.32,hs-CRP(mg/L):12.24±6.28 vs. 15.54±5.94,P<0.05 or P<0.01〕. Compared with the empirical treatment group,the time of antibiotics use(days:6.00±2.55 vs. 9.20±3.46), the duration of mechanical ventilation(days:5.00±1.73 vs. 7.00±1.94),and the length of ICU stay(days:7.43±1.72 vs. 12.57±4.16)were significantly shortened(P<0.05 or P<0.01). Conclusions The results of bedside sputum examination and sputum culture showed a good correlation,and the former is helpful in early diagnosis and treatment of VAP. The result of high quality sputum smear in significant in guiding the first choice of antibiotics,reduce the time of antibiotic use,shorten the duration of mechanical ventilation and the length of ICU stay,and improve the outcome of the patients.

AlM: To study an approach to visual acuity correction after intracapsular cataract extraction by phase - ll intraocular lens implantation through the individualized arcuate keratotomy.
METHODS: For demonstration, 48 postoperative patients ( 50 eyes ) receiving the intracapsular cataract extraction were gathered up. Each patient received a scleral tunnel major incision along the radial line of the maximum corneal refractive power determined by a cornea curvimeter, and a arcuate keratotomy was made opposite to the major one; through the major incision an iris-claw intraocular lens is implanted. Each patient was measured for their corneal astigmatism and uncorrected visual acuity before and after the surgery.
RESULTS: The results suggested the average corneal astigmatism before the surgery and that 3d, 1, 3, 6 and 12mo after the surgery as+3. 18±0. 68,-1. 56±0. 73,+0. 87± 0. 51, + 1. 21 ± 0. 70, + 1. 33 ± 0. 68 and + 1. 48 ± 0. 48 respectively. The uncorrected visual acuities 3d, 1, 3, 6 and 12mo after the surgery are 0. 5±0. 38, 0. 56±0. 23, 0. 55± 0. 24, 0. 52±0. 28 and 0. 51±0. 25 respectively.
CONCLUSlON: Phase-ll intraocular lens implantation witharcuate keratotomy is helpful to improve the postoperative visual acuity and reduce preoperative corneal astigmatism after the intracapsular cataract extraction aphakic eyes, lt is also a low-cost surgery, and easy to perform, with minor surgical injuries, particularly available for surgical visual acuity correction of the aphakic eye receiving intracapsular cataract extraction.

Objective To determine the effects of tempol(a nitroxide), in the exposure of ultraviolet-B (UVB), on cell proliferation, superoxide enzyme (SOD) activity, lipid peroxidation, and expression of matrix metalloproteinases (MMP)-1,MMP-3 in human foreskin fibroblasts in vitro. Methods Fibroblasts were irradiated by a single exposure of 36 seconds to 40 mJ/cm 2 UVB and at the same time incubated with, or without, tempol and detected twenty-four hours later. SOD activity and lipid peroxidation,as shown by accumulation malondialdehyde (MDA) were detected by biochemical assay. Expression of MMP-1 and MMP-3 (mRNA level) were examined by Semi-quantitative RT-PCR. Results 40 mJ/cm 2 UVB significantly inhibited cell proliferation rate to (84?8)% (P

Objective To investigate the optimal regimen of narrow-band ultraviolet B (NB-UVB) phototherapy in the treatment of chronic actinic dermatitis (CAD),and to analyze factors influencing treatment compliance.Methods Demographic data,results of photobiological tests,treatment parameters and clinical responses were collected from CAD patients who received NB-UVB phototherapy in Huashan Hospital affiliated to Fudan University from January 2008 to June 2015,and were reviewed retrospectively.Statistical analysis was done by using two independent samples t-test and chi-square test with SAS9.3 software to compare the clinical data between patients who completed and did not complete the NB-UVB phototherapy.Results A total of 79 CAD patients with Fitzpatrick skin type Ⅳ received NB-UVB phototherapy.Of these patients,61 (77％) completed the whole treatment,while 18 (23％) dropped out because of intolerance to the NB-UVB radiation.Among the 61 patients who completed the treatment,the average initial,final and cumulative radiation doses of NB-UVB were (0.08 ± 0.01) J/cm2,(0.32 ± 0.08) J/cm2and (5.9 ± 2.5) J respectively,and patients received (28 ± 8) times of treatment in average.When the radiation dose went up to 0.30 J/cm2,most skin lesions were cleared in 52 (85％) patients.A total of 19patients received phototesting again after the end of phototherapy.Among 16 patients sensitive to ultraviolet A (UVA) before the treatment,6 had normal minimal erythema dose to UVA (UVA-MED),and another 6 had improved UVA-MED after the treatment.Among 16 patients sensitive to UVB before the treatment,11 got normal UVB-MED and another 3 had improved UVB-MED after the treatment.Univariate analysis showed no significant differences in gender,age,duration of the disease,sensitivity to UVA and UVB radiation,results of photopatch test and patch test between the patients who completed and did not complete the treatment (all P ＞ 0.05).Conclusions The appropriate NB-UVB phototherapy for CAD patients should start at an initial radiation dose of 0.08 J/cm2 in spring and end at a final radiation dose of 0.30 J/cm2 for about 28 sessions,which can effectively reduce the photosensitivity to both UVA and UVB in CAD patients.Additionally,NB-UVB phototherapy can be applied in CAD patients of different gender,age,disease duration and photosensitive condition.

Objective To detect the selective inhibitory effects of irradiation plus adenovirusmediated horseradish peroxidase ( HRP)/indole-3-acetic acid (IAA) suicide gene system using tumorspecific and radio-inducible chimeric promoter on human hepatocellular carcinoma subcutaneously xenografted in nude mouse.MethodsRecombinant replicated-deficient adenovirus vector containing HRP gene and chimeric human telomerase reverse transcriptase (hTERT) promoter carrying 6 radioinducible CArG elements was constructed.A human subcutaneous transplanting hepatocellular carcinoma (MHCC97 cell line) model was treated with -γ-ray irradiation plus intra-tumor injections of adenoviral vector and intra-peritoneal injections of prodrug IAA.The change of tumor volume and tumor growth inhibiting rate,the survival time of nude mice,as well as histopathology of xenograft tumor and normal tissues were evaluated.Results Thirty one days after the treatment,the relative tumor volumes in the negative,adenovirus therapy,irradiation,and combination groups were 49.23 ± 4.55,27.71 :± 7.74,28.53 + 10.48 and 11.58 ± 3.23,respectively.There was a significantly statistical difference among them (F = 16.288,P ＜0.01 ).The inhibition effect in the combination group was strongest as compared with that in other groups,and its inhibition ratio was 76.5%.The survival period extended to 43 d in the combination group,which showed a significantly difference with that in the control group(x2 = 18.307 ,P ＜0.01 ).The area of tumors necrosis in the combination group was larger than that in the other groups,and the normal tissues showed no treatment-related toxic effect in all groups.However,multiple hepatocellular carcinoma metastases were observed in the liver in the control group,there were a few metastases in the monotherapy groups and no metastasis in the combination group.Conclusions Adenovirus-mediated suicide gene therapy plus radiotherapy dramatically could inhibit tumor growth and prolong median survival time.It might provide a promising therapeutic modality for hepatocellular carcinoma therapy.

Aim Toinvestigatingtheinductionof CYPs in hepatocytes or HepG2 cells by triptolide(TP) andthepossiblemechanism.Methods AfterTPtreat-ment,the expression of CYPs in rat primary hepato-cytes or human HepG2 cells was detected by real-time PCR and Western blot assays.Specific inhibitors or gene knockdown method were employed to analyze the possiblemechanism.Results Theexpressionof CYP1A2,2C7,2C11,2C12,2D2,2E1 and 3A1 in rat primary hepatocytes was induced by TP.The fold was 19,2,31,3,21,88 and 34 at 50 nmol·L-1, respectively while at 100 nmol·L-1 it was 20,5,30,23,61,83 and 38,respectively.In HepG2 cells,the expression of human CYP1A1,2B6,2C9,2C19, 2D6,2E1 and 3A4 was also induced by TP.The ac-tivities of nuclear receptor PXR and CAR were inhibi-ted.TP upregulated p53 expression,and the induction of several CYPs caused by TP was blocked when p53 wasinhibited.Conclusions TPinducesCYPsexpres-sion in rat hepatocytes and HepG2 cells.Nuclear re-ceptors may not be involved in TP induced CYPs, while the mechanism may partly attribute to p53.

The structure and properties about encoding protein of lactate dehydrogenase A from Taenia solium(Ts LDH-A)were analyzed and predicted by bioinformatics in this study.The immunological characteristics of this novel gene were also analyzed by cloning and expressing.The full-length cDNA encoding Ts LDH-A was identified from the cDNA plasmid library by blastx and rpsblast programs provided by NCBI.The physico-chemical properties and structures of Ts LDH-A were analyzed by tools provided by ExPASy.And the B cell epitopes of Ts LDH-A were predicted by the B Cell Epitope Prediction Tools provided by IEDB Analysis Resource.The PCR amplified coding region of Ts LDH-A was cloned into the prokaryotic expression vector pET-28a (+) and expressed in E.coli BL21 with IPTG induction.The immunogenicity of the purified recombinant protein was analyzed by Western Blotting.It was demonstrated that the amino acid sequence of Ts LDH-A had identity with that of LDH-A from other specie and there was a conserved LDH domain in the deduced amino acid sequence.The full-length cDNA sequence encoding Ts LDH-A included a complete open reading frame(ORF)of 1332 bp and coded to a putative protein with 331 amino acids.The molecular weight of Ts LDH-A was predicted to be 35461.1 Da and the coding protein was demonstrated to contain 3 trans-membrane regions and 4 main B cell epitopes.The active site of L-lactate dehydrogenase located at the epitope aa190-199.The 3 key residues in the catalytic site of enzyme were conserved in different species and located near to each other in spatial position.PCR,double enzyme restriction and DNA sequencing were used to identify pET28a (+)-Ts LDH-A.The recombinant protein could react with the rat's sera as well as the sera from the patients and the swine infected Taenia solium.It is clear that the full-length cDNA sequence encoding Ts LDH-A can be screened from the cDNA library of adult Taenia solium by bioinformatics analysis and can be used to investigate the structure and properties about gene and encoding protein of Ts LDH-A as well as the immunological activities of gene expression in the prokaryotic system.

Objective To explore the protective effect of folic acid supplementation in different ways on lipopolysaccharide-induced external malformations and skeletal malformations in mice.Methods The pregnant mice were divided into six groups randomly,including saline control group,FA control group,LPS group,LPS+FAig group,LPS+FAip group,and LPS+FAdw group.All dams were medicated on gestational day 8～12,and then sacrificed on gestational day 18,for each litter,the number of live fetuses,dead fetuses,resorption fetuses and external malformations of live fetuses were all counted.All fetuses were subsequently evaluated the skeletal malformations.Results A low dose of LPS injection during the second trimester resulted in external and skeletal malformations.Exencephaly and encephalomeningocele were two of the most common external malformations,skeletal malformations consisted mainly of the incompletion of supraoccipital ossification,sternal malformation and rib malformation.FA supplementation by three ways all attenuated the external and skeletal malformations.And the best protective effect was by oral administration.The incidence of external,sternal and rib malformation was all decreased.Conclusions Three ways can prevent the mice during the second trimester from the external and skeletal malformations caused by LPS injection,and the best protective effect was by intragastric administration.

ED are correlated with the severity of CAD in a degree.