To develop a high-performance liquid chromatographic method for the determination of isoguanosine in Semen Crotonis Tiglii.

Objective To determine the effects of genistein and daidzein on the proliferation and survival of the hippocampal neural cells and underlying mechanism. Methods H19-7/IGF-IR neural cell line was cultured in phenol red free DMEM absented of serum for 72h. Genistein, daidzein or 17β-estradiol was added to the culture at various concentrations. Their proliferation and protective effects on the neuronal cells were determined by BrdU and MTT assay respectively. The effect of phytoestrogens on cell cycle regulation was determined using flow cytometry. The effects of the soy isoflavones on brain-derived neurotrophic factor (BDNF) expression were determined by ELISA and RT-PCR respectively. Results It was observed that, with 72h of treatment, 20nM and 200 nM 17B-estradiol significantly promoted the neuronal cell proliferation at 33% and 36% ;20nM and 200nM genistein significantly promoted the neuronal cell proliferation at 15% and 13% ; 200nM daidzein significantly promoted the neuronal cell proliferation at 11% compared to the control (P＜0.05). Genistein and daidzein induced an significantly increase in the S phase arrest at (17.64 ± 0.43) % and (19.48 ± 1.01) % compared to the control (P ＜ 0. 05). Moreover, genistein and daidzein significantly increased the expression of mature BDNF and BDNF mRNA level (P＜0.05). The effect of genistein and daidzein on hippocampal neuronal cell proliferation was blocked by K252a, selective inhibitor of tyrosine kinase receptors. Conclusion Genistein and daidzein improved hippocampal neuronal cell proliferation and viability in vitro. The effects might be mediated by increasing in BDNF expression.

Objective To develop Z-score reference ranges for mitral valve-tricuspid valve distance (MTD) in normal fetuses from the measurements of femur length (FL),biparietal diameter (BPD) or gestational age (GA) using fetal echocardiography.Methods A retrospective cross-sectional study of 1216singleton normal fetuses was performed.The gestation age ranged from 20 to 40 weeks.Non-cardiac fetal biometric parameters included FL and BPD and GA based on menstrual age.MTD was measured in a standard apical four-chamber view.Normal Z-score ranges were developed for MTD using GA,BPD and FL as independent variables.These were accomplished by using first standard regression analysis and then weighted regression of absolute residual values for each parameter in order to adjust for inconstant variance.Results Linear regression model was the best description of the data in each case and correlations between MTD and independent variables (FL,BPD,GA) were excellent.The linear regression equations between MTD and FL,BPD,GA were:Y =0.04629 × FL-0.02868 (r =0.827) ; Y =0.04018 × BPD-0.08293(r =0.767) ; Y =0.0098 × GA-0.07075 (r =0.721),respectively.Heteroscedasticity of standard deviation(SD) with increasing independent variables also could be modeled with a simple linear regression.Linear regression equations between SD of MTD and FL,BPD,GA were:Y =0.0149 × FL-0.0095 (r =0.9947),Y =0.0126 × BPD-0.02793 (r =0.9931),Y =0.00305 × GA-0.021 (r =0.991),respectively.According to these equations,MTD Z-score =(the actual measurement of MTD-predicted MTD)/prediction SD.Conclusions Normal reference ranges and Z-scores for MTD have been provided.These normative data may be useful tools for assessment of fetal MTD,especially has potential applications in Ebstein anomaly or other malformations involved atrioventricular valves.

Objective To establish the orthotopic human pancreatic cancer model in mice and study the method to detect the tumor growth. Methods Human pancreatic cancer cell lines SW1990 in logarithmic phase was made into cell suspension and in situ injected into the envelope of pancreatic tail of BALB/c nude mice. High-frequency endoscopy ultrasound was used to observe the growth of tumor mass and its imaging characteristics were studied. Results 20 nude mice were successfully implanted, and 1 died 25 h after implantation. 14 days after implantation, the tumor of pancreatic cancer on EUS was (8.09 ± 2.61) mm3, the tumor appeared as homogeneous hypoechoic mass with clear boundary, and envelope as well as sound halo was present, and the shape was regular; there was low speed circular vasculature signal in around 30% of tumor.The tumor size increased to (12.40 ± 3.51)mm3, and the shape of 70% tumor was irregular, and some appeared as lobulated, and the tumor appeared as heterogeneous hypoechoic mass, no necrosis or liquefaction was found 28 days after implantation. There was low speed circular vasculature signal in around 70% tumors.Conclusions The orthotopic pancreatic cancer model in nude mouse can be established by in situ injection and this method is relatively ideal because it is simple and effective. The high frequency probe of endoscopic ultrasonograph is a reliable method for monitoring implanted pancreatic cancer.

Objective To investigate the change of connexin (Cx) in mesenteric artery (MA) and aorta of spontaneously hyper‐tensive rats (SHR) and normotensive rats .Methods Quantitative RT‐PCR and Werstern blot technique were used to compare the difference in the expression of Cx37 and Cx40 mRNA and protein in MA and aorta of SHR and normotensive rats .Results The level of Cx37 mRNA expression in MA from SHR was decreased compared with that of the normotensive rats (P<0 .05) ,and more significantly in aorta (P<0 .01) .Cx40 mRNA expression in MA from SHR was significantly decreased compared with that of the normotensive rats (P<0 .01) ,but no change in aorta (P>0 .05) .The expression of Cx37 protein in MA from SHR was decreased compared with that of the normotensive rats (P<0 .05) ,and more significantly in aorta (P<0 .01) .Cx40 protein expression in MA from SHR was significantly decreased compared with that of the normotensive rats (P<0 .05) ,but no change in aorta (P>0 .05) . Conclusion Hypertension may could decrease gap junctional communication in cells of MA and aorta from SHR by the downregu‐lation of the expression of Cx37 and/or Cx40 .

Objective To compare the mesothelin expressions in 3 human pancreatic cancer cell lines between in vitro and in vivo and the developing speed among the subcutaneous tumors implanted with the 3 human pancreatic cancer cell lines in nude mice.Methods The human pancreatic cancer cell lines ( SW1990,BxPC3 and PANC1 ) were cultured and then were implanted subcutaneously into left axillas of nude mice.The volumes of these subcutaneous tumors were recorded every week to estimate their developing speed.The mice implanted with SW1990 and BxPC3 cells were observed for three weeks,while the mice implanted with PANC1 cell were observed for five weeks.The Western blot method was used to measure the expressions of mesothelin in the 3 kinds of cells and subcutaneous tumors,while immunohistochemical staining was applied to determine the expressions of mesothelin in 3 kinds of subcutaneous tumors.Results The sequence of quantities of expressions of mesothelin in these cell lines in vitro were BxPC3 ＞ PANC1 ＞ SW1990,and the sequence of quantities of expressions in vivo were SW1990 ＞ BxPC3 ＞ PANC1.One handrued percent of the tumors grew out successfully,and the sequence of speeds of their growth was SW1990 ＞ BxPC3 ＞ PANC1.Conclusions The mesothelin expressions among 3 kinds of pancreatic cancer cell line are different.The developing speeds of tumors originated from different subcutaneous tumors in nude mice are also different,and there is no association between them.

Objective To measure mitral annual displacement (MAD) by M-mode echocardiography in normal second and third trimester fetuses,and assess relationships between MAD and gestational age and routine echocardiographic parameters for evaluation of left ventricular function.Methods One hundred fifty-five normal fetuses from 19 to 38 weeks of gestation were recruited in the study.MVD and tricuspid annual displacement (TAD) were measured.Early diastolic inflow velocities (E) of the atrioventricular valves were assessed by pulsed-wave Doppler(PW),and early diastolic velocities (Em) of the mitral annular were estimated by pulsed-wave tissue Doppler imaging (TDI).The ratio of E/Em was calculated.Results In normal fetuses,the MAD was (7.05 ± 1.17) mm.There was significant positive correlation between the fetal MAD and advancing gestational age ( r =0.82,P ＜0.01 ),and between MAD and E,A,Em,Am or Sm ( r =0.25,0.24,0.32,0.29 and 0.40 respectively,P ＜0.01).There was no correlation between fetal MAD and E/A,E/Em,LVEF or LVFS.The left MAD was significantly lower than the right TAD ( P ＜0.01 ).Conclusions MAD of fetuses from 19 to 38 weeks of gestation can be quantitatively measured by Mmode echocardiogram.The measurement of MAD provides a reliable quantitative standard to estimate long axis ventricular function of fetuses,which may be clinically useful in prenatal detecting early cardiac failure.

Objective To evaluate the prognostic value of blood lactate (Lac) level in sepsis patients with or without diabetes.Methods 106 patients admitted to intensive care unit (ICU) of Zhongshan Hospital Affiliated to Fudan University from April 2015 to November 2016 were enrolled. The patients with age > 18 years and the length of hospital stay > 24 hours were included. Records including blood Lac, serum creatinine (SCr), white blood cell count (WBC), platelet count (PLT), sequential organ failure assessment (SOFA) on the first day of admission; minimum oxygen index (PaO2/FiO2) in 3 days after admission; mechanical ventilation, whether there was a history of diabetes, usage of biguanides, etiology control treatment, usage of continuous renal replacement therapy (CRRT) were collected. According to the level of blood Lac patients were divided into high Lac group (Lac > 2 mmol/L) and low Lac group (Lac ≤ 2 mmol/L);based on their diabetic history, sepsis patients were divided into the diabetes group and non-diabetes group. The survival curve of each group was analyzed by Kaplan-Meier regression analysis, and the factors influencing the prognosis were analyzed by multivariate Cox regression analysis.Results There were 76 males and 30 females sepsis patients, with an average age of (68.1±14.7) years old. In the 51 patients of low Lac group, there were 7 patients who suffered from diabetes. While in the 55 patients of high Lac group, there were 12 patients who suffered from diabetes. Compared with low Lac group, high Lac group had a higher age, higher SOFA score, and a lower proportion of patients who had the treatment of etiology control (allP < 0.05). There was no significant difference of blood Lac in sepsis patients with diabetes and those without diabetes (mmol/L: 3.03±2.73 vs. 2.81±2.40,P > 0.05). Kaplan-Meier survival curve analysis showed that the 90-day survival rate in the high Lac group was significantly lower than that in the low Lac group (56.36% vs. 90.20%,χ2 = 0.697,P = 0.008). The high Lac group without diabetes had lower survival rate, and the 90-day survival rate was significantly lower than that of the low Lac group without diabetes (58.14% vs. 90.90%,χ2 = 7.152,P = 0.007); there was no significant difference in 90-day survival rate between the high Lac group and the low Lac group with diabetes (50.00% vs. 85.71%,χ2 = 0.012,P = 0.914). Multivariate Cox regression analysis showed that blood Lac was an independent risk factor for the prognosis of sepsis patients [odds ratio (OR) = 3.863, 95% confidence interval (95%CI) = 1.237-12.060,P = 0.020]. After stratification according to their diabetic history, the blood Lac was an independent risk factor for the prognosis of sepsis patients without diabetes (OR = 4.816, 95%CI = 1.407-15.824, P = 0.010), but the blood Lac had no effect on the prognosis of sepsis patients with diabetes (OR = 0.000, 95%CI =0.000-1.103,P = 0.270).Conclusions The predictive value of blood Lac on sepsis patients with or without diabetes was different. The blood Lac was related with the prognosis of sepsis patients without diabetes, while further study should be conducted for the prognostic value of blood Lac in sepsis patients with diabetes, and it's possible to increase the cut-off-point of Lac level in these patients.

Objective To explore the difference in Z-score reference range for aorta measurements, and pulmonary artery(PA)/aortic(AO)valve annular diameter ratio(PA/AO ratio)between normal and aortic stenosis(AS)fetuses and to evaluate the diagnostic value of Z-score of aorta in fetuses with AS. Methods A retrospective cross-sectional study of fetal color Doppler echocardiograms were undertook from 300 singleton normal fetuses with gestational age ranged from 16 to 40 weeks.Standardized fetal echocardiographic measurements included AO valve annular diameter and PA valve annular diameter at end-systole,and PA/AO ratio were obtained.With statistical analysis,best model was established for predicting AO and PA using biparietal diameter (BPD)and femoral length (FL)as independent variables.The correlations between standard deviation (SD)and two independent variables (BPD,FL)were analyzed,and the AO Z-score and PA Z-score of normal fetuses were calculated.One hundred and six singleton fetuses with AS were studied whose gestational age ranged from 1 5 to 40 weeks.The AO Z-score and PA Z-score of AS fetuses obtained by using above regression equation and PA/AO ratio were subsequently calculated.The AO and PA Z-scores and PA/AO ratio between normal fetuses and AS fetuses were compared.And the detection rate of AS according to AO Z-score,PA/AO ratio and AO percentile range were assessed.Results Compared with normal fetuses,the Z-score of AO and the PA/AO ratio in AS fetuses were statistically different (P <0.01).While there was no statistical difference of Z-score of PA between two groups (P >0.05).AO Z-score in 96.2% of AS fetuses were found < -3,PA/AO ratio in 95.3% of AS fetuses were found outside 95% CI.The method using AO Z-score had the highest detection rate of AS,the positive predictive value was up to 89.5%,the negative predictive value was up to 98.6%,the sensitivity,specificity and Youden index were 96.2%,96%,and 0.9223,respectively.PA/AO ratio had good but slightly lower diagnostic accuracy than AO Z-score,while AO percentile range performed poor.Conclusions Fetuses whose AO Z-score are outside ±3 and the PA/AO ratio is greater than 1 .4 at the same time are more likely to suffured from AS.Quantification of AO Z-score has important clinical value in the diagnosis of AS,and is more accurate than the percentile range in quantitative assessment of congenital heart growth and development of the fetal pathology of AS.

The preventative treatment theory is a very important part of traditional Chinese medicine. When this theory is used in the treatment of common cold disease, it focuses on the prevention of common cold disease. The preventative treatment theory is very important and can be applied in the whole process of the treatment common cold disease.