Objective To investigate the safety and efficacy of reducing pulmonary arterial pressure in patients with congeni‐tal heart disease(CHD) complicated with pulmonary hypertension(PAH) .Methods From September 2014 to January 2015 in our hospital ,34 patients with CHD complicated with PAH were selected ,all of them were treated with endothelin receptor antagonist , bosentan ,on the basis of conventional therapy .The hemodynamics observation ,cardiac function detection and six‐minute walking distance test(6 MWD)were performed after pulmonary artery pressure reducing therapy .Results After treatment ,the pulmonary vascular resistance (PVR) ,arterial oxygen saturation(SaO2 ) ,Qp/Qs and right cardiac output(CO) in these patients was (218 .30 ± 91 .25)dyn · s-1 · cm -5 ,(95 .16 ± 7 .84)% ,(1 .10 ± 0 .27) and (5 .20 ± 1 .36) L/min ,respectively ,which were better than those be‐fore treatment(P<0 .05) .There was no statistically significant difference in the pulmonary artery systolic pressure(sPAP) ,pulmo‐nary artery diastolic pressure pulse AI(dPAP) ,mean pulmonary arterial pressure(MPAP) ,heart rate(HR) and central venous pres‐sure(CVP) before and after the treatment(P>0 .05) .There was no statistically significant difference in the right ventricular diame‐ter(RVD) ,left ventricular ejection fraction(LVEF) ,three tricuspid regurgitation velocity(VP) ,three tricuspid valve transvalvular pressure difference(PGTV) ,left ventricular diastolic volume (LVEDV) and left ventricular diastolic diameter(LVEDD) before and after the treatment(P>0 .05) .After 3 months and 6 months of treatment ,6 MWD was (378 .09 ± 40 .12) and (423 .07 ± 35 .32) m respectively ,both of which were higher than those before treatment ,the differences were statistically significant (P<0 .05) .Conclu‐sion The application of bosentan therapy is safe and effective for patients with CHD complicated with PAH ,which could improve pulmonary hemodynamics and 6 MWD .

Objective To observe the changes in red cell distribution width (RDW)and plasma N -terminal pro -brain natriuretic peptide(NT -proBNP)in children with chronic heart failure due to congenital heart disease (CHD),and to investigate their correlations with cardiac function.Methods Eighty -four children with chronic heart failure and pneumonia secondary to CHD who were hospitalized in Wuhan Health Care Center for Women and Children from January to December 201 4 were enrolled.According to the modified Ross scoring system,they were divided into 3 subgroups:mild,moderate and severe heart failure subgroups.Forty -five children with pneumonia who were hospitali-zed during the same period were randomly selected as the control group.Then the levels of RDW and plasma NT -proBNP were compared in each group,and the correlation between RDW and NT -proBNP was analyzed.Results The levels of RDW and plasma NT -proBNP in children with heart failure caused by CHD were significantly higher than those of the control group[(1 5.6 ±2.2)% vs (1 3.7 ±1 .5)%,(3 252.9 ±1 061 .5)ng/L vs (67.4 ±1 7.9)ng/L, t =5.51 8,7.838,all P <0.01 ].NT -proBNP levels were gradually increased along with increasing heart failure grade, and there were statistically significant differences[(1 71 8.3 ±456.2)ng/L vs (3 371 .4 ±834.2)ng/L vs (4 681 .2 ± 1 21 4.8)ng/L,F =1 5.961 ,P <0.01 ].RDW levels also increased gradually,while there were no statistically signifi-cant differences[(1 5.2 ±1 .5)% vs (1 5.7 ±2.0)% vs (1 6.0 ±2.8)%,F =2.246,P >0.05].There was no signifi-cant correlation between RDMand NT -proBNP (r =0.1 34,P >0.05).Conclusions For children with CHD,higher RDW and NT -proBNP levels can aid in the diagnosis of chronic heart failure,and NT -proBNP level is associated with the severity of heart failure,yet it is still unclear whether RDW level can reflect the severity of heart failure.

Objective To establish the primary cat intestinal epithelial cells(IECs)culture methods and construct the cD-NA library for the following yeast two-hybrid experiment,so as to screen the virulence interaction factors among the final host. Methods The primary cat IECs were cultured by the tissue cultivation and combined digestion with collagenase XI and dispase I separately. Then the cat IECs cultured was identified with the morphological observation and cyto-keratin detection ,by using goat anti-cyto-keratin monoclonal antibodies. The mRNA of cat IECs was isolated and used as the template to synthesize the first strand cDNA by SMARTTM technology,and then the double-strand cDNAs were acquired by LD-PCR,which were subsequently cloned into the plasmid PGADT7-Rec to construct yeast two-hybrid cDNA library in the yeast strain Y187 by homologous recom-bination. Matchmaker?Insert Check PCR was used to detect the size distribution of cDNA fragments after the capacity calcula-tion of the cDNA library. Results The comparison of the two cultivation methods indicated that the combined digestion of colla-genase XI and dispase I was more effective than the tissue cultivation. The cat IECs system of continuous culture was established and the cat IECs with high purity were harvested for constructing the yeast two-hybrid cDNA library. The library contained 1.1× 106 independent clones. The titer was 2.8 × 109 cfu/ml. The size of inserted fragments was among 0.5-2.0 kb. Conclusion The yeast two-hybrid cDNA library of cat IECs meets the requirements of further screen research,and this study lays the foundation of screening the Toxoplasma gondii virulence interaction factors among the cDNA libraries of its final hosts.

OBJECTIVETo explore the relationship of CD44 expression or glycosylation and hepatocellular carcinoma (HCC) metastasis.

METHODSIHC, Quantum dots detection, RT-PCR, Western blot, Cellular immune fluorescence and MS-PCR were used to identify CD44 expression in HCC samples and a series of human HCC cell lines with different metastatic potentials. Lectin array was used to reveal the relationship of CD44v6 glycosylation and human HCC metastasis.

RESULTSImmunohistochemistry analysis showed that CD44v6 was mainly distributed on the cell membrane, while CD44S immunoreactivity was prominently in the cytoplasm, CD44v3 and CD44v4/5 were in cytoplasm on membrane. Among CD44S and those CD44 variants, only the expression of CD44v6 was higher in metastasis HCC samples as compared to that in non-metastasis group (x²=8.828, P less than 0.05). This result was also re-confirmed by the result of Quantum dots (t = 2.392, P < 0.05) and serum detection (t = 2.56, P < 0.05). We found completely methylation of CD44v6 gene in Hep3B and incomplete methylation in MHCC97H and MHCC97L cell lines with metastatic potentials. The lectin affinity assay indicated that lectin MAL, SNA and WGA showed more affinity to MHCC97H and MHCC97Lcell lines than that of the non-metastatic Hep3B cell lines.

CONCLUSIONSCD44v6 over-expression presents a positive correlation with HCC metastatic potential, which may be associated with DNA methylation level in promoter sequence. The increasing sialic acid modified glycan of CD44v6 might be related to HCC metastatic ability.

Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; DNA Methylation ; Glycosylation ; Humans ; Hyaluronan Receptors ; metabolism ; Liver Neoplasms ; metabolism ; pathology ; Neoplasm Metastasis ; pathology ; Promoter Regions, Genetic

Objective To evaluate the role of contrast-enhanced ultrasound (CEUS) on the the detection of hepatic metastases in comparison with conventional ultrasonography (US) and contrastenhanced computed tomography (CECT).MethodsNinety-seven patients with hepatic metastases underwent US,CEUS and CECT images.Their detection of hepatic metastases were compared.Results Hepatic metastases showed five enhancement patterns with CEUS,including bolus hyper-enhancement,peripheralrimenhancement,inhomogeneousenhancementwithnecrosis,hypo-enhancement,isoenhancement with liver parenchyma,all hepatic metastases showed dark defects in portal and delayed phase.The mean number of metastases at CEUS was greater than that of US (2.6±1.9 vs 1.6 ±1.2,P＜0.05).The detection of hepatic metastases was 53.4％ and 87.1％ respectively(P＜0.05).CEUS and CECT have no statistically significant difference in the detection of hepatic metastases (P＞0.05).ConclusionsCEUS can improve detection of hepatic metastases.

Objective To investigate the effect of functional training on knee pain,functional movement screen (FMS) score and balance in Chinese elite fencing athletes with patellar tendinopathy.Methods Twenty-four fencing athletes with a diagnosed patellar tendinopathy were randomized into a treatment group (TG) and a control group (CG),each of 12.Both groups were given routine physical therapy,while TG received motor function training in addition for eight weeks.Both groups completed the numerical rating scale (NRS),FMS and balance test before and after the intervention.Results After the intervention,the average PRS and FMS of TG were 2.08± 1.24 and 16.25±0.97 respectively,which significantly outperformed those of TG before the intervention and those of CG after the intervention (P＜0.05).Moreover,TG indicated superior results in parameters of static postural balance including center of pressure,total length of swinging pathway,maximal length of swinging pathway,and area of swinging pathway when compared to TG before the intervention and CG after the intervention(P＜0.05).Conclusion The motor functiontraining is effective in improving functional movement and balance in elite fencing athletes with patellar tendinopathy.

Objective To explore pathological characterization of the enlarged contrast enhanced area of hepatocellular carcinoma ( HCC) on contrast enhanced ultrasonography ( CEUS). Methods Thirty-five HCC patients underwent conventional ultrasonography and CEUS before hepatectomy. The tumor sizes were measured on conventional ultrasonography,CEUS and postoperative resected specimen at the largest section. The infiltration depth of the HCC was observed under microscope in the peripheral area of the tumors. HCC with the size of contrast enhancement area on CEUS 0. 3 cm larger than that on conventional ultrasonography was defined as group A, and the others as group B. The pathological character of the tumor and tissues surrounding the tumor were analyzed and compared between the two groups. Results Among 35 HCC, 40% (14/35) HCC increased in size on CEUS compared with ultrasonography. Of them,9 tumors with increased area on CEUS were confirmed to be invasive HCC on pathology. Using the enlarged contrast enhanced area on CEUS as the diagnostic criteria of invasive HCC, the sensitivity, specificity, accuracy, positive predictive value and negative predictive value were 90%, 80%, 82.9%, 64.3%, 95. 2%, respectively. Conclusions The contrast enhanced area on CEUS can reflect the infiltrative tumor size more accurately compared with conventional ultrasonography.

Objective To explore the dynamic changes and clinical significance of relative pituitary hormones in children after craniocerebral injury.Methods The quantitative analysis and dynamic observation were performed in 125 children after craniocerebral injury and 20 voluntary healthy children of relative pituitary hormones including serum prolactin(PRL),cortisol(PTC),three free triiodothyronine (FT3),free thyroxine (FT4),thyroid stimulating hormone (TSH),growth hormone (GH) by applying electrochemical luminescence method.Tbe hormone variational characteristics were analyzed according to posttraumatic time,Glasgow Coma Scale(GCS) scores on admission and Glasgow Outcome Scale (GOS) scores on discharge,and the relationship between hormone variational characteristics of 58 cases was followed up over 2 years and the activities of daily living (ADL) were also investigated.Results The serum PRL was significantly increased on the first,third and fifth day compared with the healthy control group (P =0.000 0,0.000 0,0.006 7),respectively.There was significant difference between mild,moderate and severe groups within 30 days after suffering from craniocerebral injury (P ＜ 0.05).PTC was heavily increased within 3 days,and significant difference existed among mild and moderate groups mild and severe groups (all P ＜ 0.05) ; TSH,FT3,FT4 decreased slightly after injury and gradually rose in later;GH change wasn't significant;and the larger variation of relative pituitary hormones was responsible for lower GCS scores;FT3,FT4,TSH,and GH decreased in different degrees,which were found in parts of children with craniocerebral injury,and the significant difference of serum PRL existed between GOS scores 4-5 and GOS scores 1-3 groups (P =0.000 1).Conclusions The changes of relative pituitary hormones were associated with the posttraumatic time and the severity of craniocerebral injury.The PRL in serum can aid in prediction of outcome for the children with craniocerebral injury.

Objective To identify genes differentially expressed in the window of implantation and explore the molecular basis of the development of endometrial receptivity.Methods A subtracted cDNA library of the window of implantation was constructed by suppression subtractive hybridization(SSH) method.The screened clones of the subtracted library were sequenced and GenBank homology search was performed.The differential expression of ribosomal protein(RP)L7,RPL7 pseudogene(RPL7p),RPL19 and tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein zeta polypeptide(YWHAZ) were further confirmed by RT-PCR.Results After sequencing and GenBank homology search of 50 clones, 35 differentially expressed genes were detected in the window of implantation,of which 23 were known genes,and 12 were unknown genes.Some of the known genes have been proved to be associated with implantation,while others were firstly screened out by us.The results of RT-PCR confirmed that RPL7, RPL7p,RPL19 and YWHAZ were highly expressed in the window of implantation,0.75?0.21,1.72? 0.30,1.23?0.31,and 1.28?0.08,respectively.Conclusions SSH is a useful technique to detect differential expression genes and an effective method to clone novel genes.It provides a new method to investigate the molecular basis of the development of endometrial receptivity.

AIM: To study cellular and molecular mechanisms of cardiac development associated genes ex- pression and its function during early stage cardiomyogenesis. METHODS: (1 ) Mouse embryonic stem cells (ESC) line D3 culture. (2) Inductive culals of ESC differentiated into cardiac myocytes in vitro.(3) Identification of ESC -derived cardiac myocytes: RNA isolation; synthesis of specific primer and RT - PCR; Label of RT - PCR products with [? - 32P] dATP as probes, purifyed by sephadex G - 50 columns, determined the yield of DNA. RNA dot hy- bridization. RESULTS: 80% of ESC differentiated into cardiomyocytes by improved conditional medium. Cardiomy- ocytes contraCted in a synchronous manner. The results of RT - PCR and RNA blot showed that cardiac genes were expressed abundantly and specifically during the early cardiomyogenesis. CONCLUSIONS: ESC were able to be dif- ferentiate into cardiomyocytes. Different concentrations and components of RA, DMSO and FCS affected ESC car- diomyogenesis in de. The optimal result obtained was from the conditional medium, a mixturce of 2 nmol/L retinoic acid (RA), 0.6% dimethyl sulfoxide (DMSO) and 20% fend calf serum (FCS).