AIM: To explore the effects of PMA(phorbol-12-myristate-13-acetate,a tumor promoter, mimicking the action of diacylglycerol on PKC)and laminin on the adhesion and the proliferation of human hepatocellular carcinoma cells, and provide a new clue to liver cancer treatment. METHODS: Human hepatocellular carcinoma cell line(BEL-7402)was used to identify the endogenous laminin and protein kinase C-?(PKC-?) expression, and the effects of laminin and PMA on the adhesion and the proliferation were also investigated in vitro. RESULTS: By the effect of exogenous laminin, human hepatocellular carcinoma cell (BEL-7402) possessed endogenous laminin expression and increased the adhesion and the proliferation, which was showed the synergistic action by the effect of PMA in combination. By the action of PMA alone,the proliferation and the PKC-? expression increased by exogenous laminin were decreased, and the adhesion and the endogenous laminin expression were increased. CONCLUSIONS: The finding suggested that the adhesion and the proliferation of human hepatocellular carcinoma cell were closely related to the effects of endogenous or exogenous laminin ,which were associated with cPKC-? activity. Therefore,the application of anti-laminin antibody in combination with PKC antagonist might be a new clue to find out the therapy for liver cancer.

Objective To detect quantitatively hepatocyte growth factor(HGF)mRNA expressions of bone marrow mononuclear cells(MNCs)in acute leukemia(AL)and investigate its clinical significance.Methods Total mRNA of quantitated bone marrow MNCs isolated from 67 de novo AL cases was extrated and then cDNA was synthesized.Expression of HGF mRNA was quantified absolutely using real-time fluorescence quantification PCR(FQ-PCR).Results Expressions of HGF mRNA in a group of AL were higher significantiv than these in a control group(6.936 ±1.613,0.407 ±0.170,P<0.001),but there was similafitv between a group of acute myeloid leukemia(AMI,)and group of acute lymphoblastic leukemia (ALL)(7.127±1.911,6.635±0.934,P>0.05).In AL subtypes,the expression of M5(9.998 4±1.454)was higher than that of M2,M3,M4,L1,L2 and L3(P<0.001),but there ware no differences among the latters(P>0.05). Meanwhile,there was no statistical significance on the expressions of HGF mRNA between different age and sex(P>0.05).In addition,expressions of HGF mRNA in the remission group were lower than these in the non.remission group(6.393±1.165,8.041±1.848,P<0.005).Conclusions There are statistical significances of the expressions of bone marrow MNCs HGF mRNA among the AL group and control group.As to AL subtypes,there are no statistically significant differences between AML and ALL as well as between different age and sex.Besides,lower HGF mRNA level is correlated with better curative effect.It is suggested that HGF mRNA is a suitable index for AL diagnosis and treatment.

Objective To construct quantitative standard for quantification of hepatocyte growth factor (HGF) mRNA and establish its real-time fluorescence quantitative(FQ)-PCR assay to estimate its clinical relevance in lymphoma.Methods Recombinant plasmid Was constructed with target cDNA obtained from isolated total RNA by RT-PCR After PCR products were identified and purified,recombined plasmids were quantitated and then acted as quantitative standard.A new real time FQ-PCR analysis system Was established with the second pair of primers and the probe after amplification condition and the concentrations of components were optimized.HGF mRNA expressions in 47 lymohoma cases[11 Hodgkin disease(HD) cases,36 non-Hodgkin lyphoma(NHL)cases.among these patients,36 patients in remission while 11 patients without remission ] were analyzed quantitatively,and its specificity and sensitivity for lymphoma diagnosis were evaluated by receiptor operation character(ROC)curve method.Results HGF mRNA quantitative standard was constructed successfully.and its real time FO.PCR analysis system Was established combined with hot.start PCR and down.touch PCR technique. According to slope of standard curve (-3.513)and correlation cofficient(0.999),amplification efficiency of the system was 92.6%.Coefficient variation of intra-assay,intra-day and inter-day-assay were 2.1%,4.0% and 6.8%,respectively.Sensitivity of FQ-PCR Was 2 eopies/μl.Expressions of HGF mRNA in lymphoma group Was higher than that in control group(6.425±2.172 and 0.317±0.192,respectively,t=15.883,P<0.001),and its expressions in remission group was lower than no remission group(6.157±1.712 and 7.59l ±1.184,respectively,t=2.768,P<0.05).However,there Was not difference of HGF mRNA level between group HD and group NHL(P>0.05).According to ROC analysis,its sensitivity and specificity were 93.6% and 100% when cutoff value for lymphoma clinical diagnosis Was 3.136.Conclusion HGF mRNA'8 quantitative standard and its real time F9-PCR analysis system have been successfully constructed,and it can be used for quantitative detection of its mRNA expression in lymphoma.

Objective To investigate the value of echocardiography application in the diagnosis of large amount of pericardial effusion (PE) in children by retrospective etiology and prognostic analysis.Methods A total of 68 children with large amount of PE admitted in Beijing Children's Hospital,Capital Medical University from March 2010 to March 2016 were reviewed retrospectively,and the echocardiographic presentations,etiology diagnosis,treatment,and prognosis were analyzed.Results Analysis indicated that the etiology diagnosis ratio was 97.1％ (66/68 cases),the effective ratio of the medical treatment was 71.8％ (46/64 cases),and the mortality rate was 7.8％ (5/64 cases).The morbidity was ranked from high to low as follow:large amount of PE included infectious pericarditis 27 cases,tumor associated PE 14 cases,postpericardiotomy syndrome and cardiotrauma 14 cases,systemic diseases 6 cases,vasa lymphatic related diseases 5 cases and idiopathic PE 2 cases.PE resulted from tumor associated PE and vasa lymphatic diseases had lower cure rate than the overall cure rate,and there were statistical significance (28.6％ vs.71.8％,x2 =9.092,P =0.003;20.0％ vs.71.8％,x2 =5.059,P =0.018).Thirty-five children received pericardiocentesis or fenestration drainage.Chylopericardium was restricted to vasa lymphatic diseases.All tumor associated PE had the hemorrhagic PE.The strong echo point in the pericardial cavity might indicate implication for hemorrhagic PE (x2 =5.166,P =0.023).The abnormal mass detected by echocardiography might indicate the tumor related or infectious causes.Conclusions The etiology of large amount of PE could be determined in most of children.As the preferred diagnosis method,echocardiography could help etiological diagnosis combined with clinical manifestation.

Objective To investigate the characteristics of transperineal pelvic ultrasounds among middlelate gestational women and study the the correlation between middle-late gestation and stress urinary incontinence.Methods Seventy-four pregnant women at a gestation of 14-40 weeks were divided into the incontinence group (n =34) and the non-incontinence group (n =40).The bladder neck angle,the bladder neck rotation angle,the bladder neck movement degrees,the posterior urethra-vesical angle,and the urethral rotation angle at both inactivity and maximum Valslva activity were evaluated by pelvic ultrasound.The acquired data were treated and analyzed with t-test and correlation analysis with with P ＜ 0.05 considered statistically significant.Results Compared with the non-incontinence group,the bladder neck rotation angle,BND,bladder neck angle and posterior urethra-vesical angle during the maximum Valsalva activity in the incontinence group were all significantly larger (P ＜ 0.05).A moderate correlation between urethral rotation angle and gestation week in the incontinence was observed (P ＜ 0.05).Conclusion The changes of the supportive structures around bladder urethreal tissues in middle-late gestation women are obvious.The urethral rotation angle becomes larger with the gestation going on.

Objective:To establish cell lines stably expressing Rab5a and its the inactive mutant Rab5aN133I,analyze the effect of Rab5a on the expression of cytokines in LPS-stimulated RAW264.7 cells .Methods:RAW264.7 cells were transfected with Rab5a and its inactive mutant vector Rab5a N133I separately,and then screened by G418.Rab5a stable expressing cell lines were identified by Real time-PCR.The growth of the stable cell lines was analyzed by MTT assay.After the stable cell lines were stimulated by LPS for different time periods,the expression of iNOS,TNF-αand IL-6 was detected.Results:Rab5a and Rab5aN133I transfection resulted in elevated Rab5a mRNA expression compared with the control cells ( P<0.05 ).Rab5a overexpression enhanced the proliferation of RAW264.7 cells.However,the proliferation of Rab5aN133I cells was significantly slower than the control cells ( P<0.05).Overexpression of Rab5a promoted LPS-induced production of iNOS,TNF-αand IL-6 in RAW264.7 cells (P<0.01). Conversely,overexpression of Rab5aN133I abolished the stimulating effects of Rab5a.Conclusion: Rab5a promoted LPS-induced expression of iNOS,TNF-αand IL-6 in RAW264.7 macrophages in a GTP-binding ability-dependent manner.

To investigate the effects of 2-(4-methoxycarbonyl-2-tetradecyloxyphenyl)carbamoylbenzoic acid (CX09040) on protecting pancreatic β cells, the β cell dysfunction model mice were induced by injection of alloxan into the caudal vein of ICR mice, and were treated with compound CX09040. Liraglutide was used as the positive control drug. The amount and the size of islets observed in pathological sections were calculated to evaluate the β cell mass; the glucose stimulated insulin secretion (GSIS) test was applied to estimate the β cell secretary function; the oral glucose tolerance test (OGTT) was taken to observe the glucose metabolism in mice; the expressions of protein in pancreas were detected by Western blotting. The effects on the target protein tyrosine phosphatase 1B (PTP1B) were assessed by the PTP1B activities of both recombinant protein and the intracellular enzyme, and by the PTP1B expression in the pancreas of mice, separately. As the results, with the treatment of CX09040 in alloxan-induced β cell dysfunction mice, the islet amount (P<0.05) and size (P<0.05) increased significantly, the changes of serum insulin in GSIS (P<0.01) and the values of acute insulin response (AIR, P<0.01) were enhanced, compared to those in model group; the impaired glucose tolerance was also ameliorated by CX09040 with the decrease of the values of area under curve (AUC, P<0.01). The activation of the signaling pathways related to β cell proliferation was enhanced by increasing the levels of p-Akt/Akt (P<0.01), p-FoxO1/FoxOl (P<0.001) and PDX-1 (P<0.01). The effects of CX09040 on PTP1B were observed by inhibiting the recombinant hPTP1B activity with IC50 value of 2.78x 10(-7) mol.L-1, reducing the intracellular PTP1B activity of 72.8% (P<0.001), suppressing the PTP1B expression (P<0.001) and up-regulating p-IRβ/IRβ (P<0.01) in pancreas of the β cell dysfunction mice, separately. In conclusion, compound CX09040 showed significant protection effects against the dysfunction of β cell of mice by enlarging the pancreatic β cell mass and increasing the glucose-induced insulin secretion; its major mechanism may be the inhibition on target PTP1B and the succedent up-regulation of β cell proliferation.

Objective:To observe any effect of intermittent theta burst stimulation (iTBS) on the spatially-delayed responses of working memory using cynomolgus macaques.Methods:The working memory of six male cynomolgus macaques (8-9 years old) was trained using a spatially-delayed response task. They were then randomly divided into an iTBS group and a control group, each of 3. The iTBS group was given iTBS at an intensity of 35% of the maximum output, with 2 seconds of stimulation followed by 8 seconds of rest with trains of 50Hz bursts repeated at a frequency of 5Hz over a period of 192 seconds once daily for 5 days, while the control group was given sham iTBS. Before and after the 5 days, the body weight and working memory of each animal were evaluated. The total number of effective feeding episodes, and of effective feeding episodes with short and long delay periods were recorded.Results:There was no significant change in the average body weight of either group before and after the modeling and iTBS intervention. After the intervention the number of total effective feeding cases and those with a short delay period were both significantly higher in the iTBS group than in the control group. However, no significant inter-group differences in the effective feeding cases with a long delay period were observed.Conclusions:iTBS is effective in improving the spatially-delayed responses of working memory, at least in cynomolgus macaques.

Objective:To investigate the influencing factors of the survival in infants with congenital diaphragmatic hernia (CDH) and to develop a prediction model for CDH.Methods:Clinical data of 252 infants with a prenatal diagnosis of CDH in Xinhua Hospital Affiliated to Shanghai Jiaotong University School of Medicine from January 2010 to December 2018 were retrospectively analyzed. Adverse outcomes were defined as neonatal death on discharge. Chi-square and t test were used for univariate analysis of 16 perinatal and five postnatal risk factors. Regression analysis was used to determine the independent predictors of survival. The receiver operating characteristics (ROC) curves of the risk factors for predicting the survival of CDH were drawn. A prediction model based on the combination of risk factors for predicting adverse outcomes of CDH was established using the cut-off value. ROC curves of the model were drawn and the area under curve (AUC), sensitivity, and specificity were calculated. Results:Out of the 252 patients, 173(68.7%) survived on discharge. Univariate analysis showed that lung-to-head ratio (LHR), polyhydramnios, right diaphragmatic hernia, liver herniation, intrathoracic stomach, premature birth, low birth weight, 5 min Apgar score <7, and amniotic fluid index (AFI) were significantly associated with the survival of CDH. Regression analysis showed that polyhydramnios ( OR=11.19,95% CI:2.83-45.33), liver herniation ( OR=2.81,95% CI:1.32-11.92), intrathoracic stomach ( OR=5.02, 95% CI:1.29-17.13), low birth weight ( OR=8.58,95% CI:1.59-45.01) and AFI ( OR=3.68, 95% CI:1.37-14.72) were independent risk factors for survival at discharge in children with CDH, while LHR ( OR=0.36, 95% CI:0.01-0.69) were protective factors. The predictive cut-off values of LHR, polyhydramnios, liver herniation, intrathoracic stomach, low birth weight, and AFI were 1.6, 1.0, 1.0, 1.0, 1.0, and 232.5 mm, respectively. The model based on the combination of the above indicators for predicting CDH adverse outcomes was shown with an AUC value of 0.904, predictive sensitivity of 0.747, and specificity of 0.896. Conclusions:In this study, LHR, liver herniation, intrathoracic stomach, polyhydramnios, low birth weight, and AFI are independent risk factors for CDH survival. The combination of prenatal and postnatal indicators is noted for a higher accuracy for predicting CDH survival.

Objective To investigate the diagnosis and prognosis of patients with coronary artery aneurysm in Kawasaki disease.Methods The data of ultrasonic diagnosis and follow-up of 338 children with Kawasaki disease complicated with coronary artery aneurysms were analyzed.Results The incidence of coronary artery aneurysm in the acute stage of Kawasaki disease was 21.2％ (338/1 594).Of all the 338 cases,small aneurysms was 66.6％ (225 cases),medium aneurysms was 25.1％ (85 cases),and giant aneurysms was 8.3％ (28 cases).There were 719 branches involved in 338 cases,32.8％ of them in left main coronary artery and 31.1％ in right coronary artery;25.3％ in left anterior descending branch,and 10.8％ in left circumflex branch.A total of 382 branches were followed up,including 218 branches of small coronary aneurysm group and 82.1％ of them were completely recovered to the normal diameter.The medium aneurysm group was 124 brunches,the proportion of no significant change,retraction,and normal were 23.4％,68.5％ and 8.1％,respectively.There were 40 brunches in the giant anuerysm group,in which the proportion of no significant change,retraction,and normal were 70.0％,27.5％ and 2.5％,respectively.A total of 30 thrombosiswere detected by echocardiography in the acute stage,20 thrombosis were regularly followed up,12 thrombosis gradually subsided,and the other remained persist.Five thrombosis were detected in the sequelae stage,and all located in the giant coronary aneurysm.Seven children were clinically diagnosed with ischemic heart disease,of them,acute myocardial infarction in 1 case,1 died of heart failure.Conclusions Kawasaki disease coronary artery disease are common in small coronary aneurysms,of which the left main artery and right coronary artery lesions are the most common,and the prognosis is better;medium and giant aneurysm need more time to recovery,and are easy to complicated with thrombosis.