Objective The purpose of this study was to investigate the effect of a dietary modification intervention model applied by ward nurse on change of dietary behavior among patients with type 2 diabetes mellitus (T2DM).Methods A total of 80 participants were divided into intervention patients (n=40) and control subjects (n=40) by random number table.Except lecture-based diabetes educational which was applied for control subjects,a dietary modification intervention model was conducted in intervention patients for a period of two weeks.The intervention program consisted of evaluating an individual's stage of change after being provided dietary information regarding kind of food and portions,discussion with a role model,and keeping a food diary record.Body mass index (BMI),waist-hip ratio (WHR),fasting plasma glucose (FPG),postprandial 2-h plasma glucose (2hPG),glycosylated hemoglobin (HbAlc) and score of healthy eating behavior were measured at initial and six months later.Results Compared with control group,BMI,WHR,FPG,2hPG,HbA1c in intervention group were significantly decreased,P ＜ 0.01 or 0.05.After six months intervention,FPG,2hPG and HbA1c in both groups were significantly decreased compared with baseline levels,P＜ 0.01.Compared with control group,the scores of healthy eating behavior in intervention group were significantly decreased,P＜ 0.05.After six months intervention,the scores of healthy eating behavior in both groups were significantly elevated,P ＜ 0.01,compared with baseline levels.Conclusions This study yielded evidence for the benefits of using the dietary modification intervention model as a framework in healthy eating behavior among patients with T2DM.

Objective To study the status of mouse fetal transgenic cells existed in the maternal blood in peri-natal period and provide the information for gene diagnosis using fetal cells circulated in the maternal blood.Methods Mating the female normal mice with male transgenic mice integrated with GFP reporter gene driven by ?-globin promoter and HS2-HS3 elements of LCR. Around the offspring were born, maternal blood was collected and GFP expression level was determined with FACS analysis. Meanwhile, DNA extracted from the tails of the mothers and their offspring as well as the maternal blood were analyzed by PCR.Results GFP positive fetal cells were not found in maternal blood before offspring were born. 1-3 weeks later, GFP positive fetal cells were detected and this population in maternal peripheral blood reached the peak. 4-5 weeks later, they disappeared gradually. PCR results showed no GFP positive band in the mothers. However, a positive fragment of GFP gene was amplified in maternal blood samples after their offspring were born. This result was in accordance with FACS analysis.Conclusion Transgene cells may be useful markers for the study of status of fetal cells existed in the maternal circulating blood. The results would be beneficial for the gene diagnosis using maternal blood as an alternative resource of fetal cells.

We established methods to isolate human amniotic fluid-derived progenitor cells (hAFPCs), and analyze the ability of hAFPCs to secrete human coagulation factor IX (hFIX) after gene modification. The hAFPCs were manually isolated by selection for attachment to gelatin coated culture dish. hFIX cDNA was transfected into hAPFCs by using a lentiviral vector. The hFIX protein concentration and activity produced from hAFPCs were determined by enzyme-linked immunosorbent assay (ELISA) and clotting assay. The isolated spindle-shaped cells showed fibroblastoid morphology after three culture passages. The doubling time in culture was 39.05 hours. Immunocytochemistry staining of the fibroblast-like cells from amniotic fluid detected expression of stem cell markers such as SSEA4 and TRA1-60. Quantitative PCR analysis demonstrated the expression of NANOG, OCT4 and SOX2 mRNAs. Transfected hAFPCs could produce and secrete hFIX into the culture medium. The observed concentration of secreted hFIX was 20.37% +/- 2.77% two days after passage, with clotting activity of 16.42% +/- 1.78%. The amount of hFIX:Ag reached a plateau of 50.35% +/- 5.42%, with clotting activity 45.34% +/- 4.67%. In conclusion, this study established method to isolate and culture amniotic fluid progenitor cells. Transfected hAFPCs can produce hFIX at stable levels in vitro, and clotting activity increases with higher hFIX concentration. Genetically engineered hAFPC are a potential method for prenatal treatment of hemophilia B.
Amniotic Fluid ; cytology ; Blood Coagulation ; Cell Culture Techniques ; Cell Separation ; methods ; DNA, Complementary ; Factor IX ; biosynthesis ; Genetic Engineering ; Genetic Vectors ; Humans ; Stem Cells ; cytology ; metabolism ; Transfection

Objective To establish the methodology of flow cytometry for detecting human cells in human/goat chimerisra.Methods Human hemopoietic stem/progenitor cells (CD+34 cells) or MIG-tranadueed-GFP CD+34 cells were transplanted into the peritoneal cavity of fetal goats in utero to obtain human/goat chimera modeL The peripheral blood cells from the chimeras were labeled with multiple mouse anti-human antibodies and the monoelonal antibodies that were specific for human but had not or only minimal cross-reaction with goat were screened as the primary antibodies for routine analysis in flow cytometry.Human cord blood was proportionally (25% ,50% ,75%,100%) added into the blood of the untransplanted goats and the cells were labeled with CD+34 monoclonal antibody.The region and size of the "gate" were chosen based on to the distribution of CD+34 cells or human cord blood.One human/goat chimera marked with GFP (MIG goat) was sacrificed and the substantial liver cells from its perfused liver were analyzed for the GFP+cells percentage and DNA contents by flow cytometry.Results CD7,CD15,CD38,CD45CD20CD34CD14and GPA monoclonal antibodies were chosen as the primary antibodies in rou tine detection by flow cytometry.The size and area of the "gate" were also defined.29.1% (29100/100 000 ) of the substantial liver cells from the MIG goat expressed GFP.DNA content analysis showed that the GFP+ cells obtained from the liver of MIG goat mainly manifested two peaks that were correspond to those of human.Conclusions Flow cytometry is rapid,simple and effective for the investigation of differentiation,homing and biological characteristics of stem cells in vivo.The selections of suitable surface antibodies and the "gate" are very important for detecting human cells accurately in the human/goat chimerism.

A reporter system for φC31 integrase was developed in NIH3T3 cells. The reporter plasmid coding green fluorescent protein (GFP) coupled with red fluorescent protein (RFP) was eo-transfected with the plasmid coding φC31 integrase, to show the activity of integrase in the cells. Fluorescence activated cell sorter (FACS) was used to measure the proportion of the cells containing red and green fluorescence. The increment of green cells was positively related to the increase in the transfection with plasmid coding φC31 integrase. Approximately 90% of green cells were observed under a ratio of [plasmid-φC31-integrase]/[reporter plasmid] at 10 : 1. This suggests that the φC31 integrase reporter system provides a probe for the function of φC31 integrase in cells.

A reporter system for ?C31 integrase was developed in NIH3T3 cells.The reporter plasmid coding green fluorescent protein(GFP) coupled with red fluorescent protein(RFP) was co-transfected with the plasmid coding ?C31 integrase, to show the activity of integrase in the cells.Fluorescence activated cell sorter(FACS) was used to measure the proportion of the cells containing red and green fluorescence.The increment of green cells was positively related to the increase in the transfection with plasmid coding ?C31 integrase.Approximately 90% of green cells were observed under a ratio of plasmid-?C31-integrase/reporter plasmid at 10∶1.This suggests that the ?C31 integrase reporter system provides a probe for the function of ?C31 integrase in cells.

Objective To investigate the value of quantitative analysis of the left ventricular longitudinal strain in patients with hypertrophic cardiomyopathy (HCM) and with normal left ventricular ejection fraction (LVEF) by using two-dimensional speckle tracking imaging.Methods Twenty-eight HCM patients with normal LVEF (all of the cases were non obstructive HCM),who were diagnosed by clinical and ultrasound echocardiography between January 1,2015 and January 1,2016 in the First Affiliated Hospital of Dalian Medical University,served as the experimental group.And twenty healthy volunteers served as the healthy control group.The peak longitudinal strain (LPS) of the left ventricle and the systolic peak of the left ventricle were calculated by the STE technique.The indexes such as the transmural gradient (△ LS=LPSEndo-LPSEpi)and the transmural gradient percentage (△ LS％=△ LS/LPSEndo) were calculated.The Peak systolic longitudinal strain of endocardium (LPSEndo),the peak systolic longitudinal strain of mid-cardium (LPSMid),the peak systolic longitudinal strain of epicardium (LPSEpi),the peak systolic longitudinal strain of basal segment (LPSb),the peak systolic longitudinal strain of middle segment (LPSm),the peak systolic longitudinal strain of apical segment (LPSa),the global peak systolic longitudinal strain (GLPS) and other left ventricular myocardial strain,such as △ LS,△ LS％,in both the HCM group and the healthy control group,were analyzed by using independent samples t test comparison.For each layer of the left ventricle and the overall myocardial longitudinal strain,two independent sample t test was used for comparison between groups,and LSD-t test was used for intra-group comparison.Results (1) There was a gradient of LPS among the three layers and the three segments in both of the two groups:LPSEndo and LPSMid [(18.36±4.97)％ vs (13.80±4.23)％,(26.41±2.93)％ vs (22.19±2.49)％],the difference was statistically significant (t=5.550,8.529,P ＜ 0.05);LPSEndo and LPSEpi [(18.36±4.97) ％ vs (11.91 ±3.63)％,(26.41±2.93)％ vs (19.43±2.20)％],the difference was statistically significant (t=5.550,8.529,P ＜ 0.05);There was significant difference between LPSMid and LPSEpi in the healthy control group [(22.19 ± 2.49)％ vs (19.43 ± 2.20)％,t=3.709,P ＜ 0.05)],that was,LPSEndo ＞ LPSMid ＞ LPSEpi.LPSa and LPSm,the difference was statistically significant (t=4.029,6.839,P ＜ 0.05);LPSa and LPSb,the difference was statistically significant (t=5.304,9.887,P ＜ 0.05);There was significant difference between LPSm and LPSb in the healthy control group (t=4.170,P ＜ 0.05);that was,LPSa ＞ LPSm ＞ LPSb.In the HCM group,LPS in the 3 layers,3 segments,and the whole left ventricular wall were lower than that of the the healthy control group,the differences were statistically significant [GLPS:(14.63± 3.75)％ vs (22.68±2.51)％,t=-8.347;LPSEndo to LPSEpi:t=-6.477,-7.909,-8.242;LPSa to LPSb:t=-6.647,-8.790,-7.267;all P ＜ 0.05).(2) Compared with the healthy control group,both the segmental gradient and global transmural gradient in the HCM group were found reduced,but the difference had no statistical significance (all P ＞ 0.05).(3) The transmural gradient percentage both in the healthy control group and the HCM group were reduced from the apical segment to the basal segment,the difference were statistically significant (HCM group:t=9.985,5.969;healthy control group:t=17.513,7.043;all P ＜ 0.05).Compared with the healthy control group,the △ LS％a and the △ LS％m of HCM group were significantly higher [(58.86± 11.32)％ vs (43.70±4.73)％,(28.43± 11.48)％ vs (20.30± 3.66)％],and the difference was statistically significant (t=5.634,3.049,all P ＜ 0.05).Conclusions (1) Using 2D-STI could accurately determine the regional or the global left ventricular systolic function in patients with HCM.(2) The transmural gradient percentage can be more sensitive to reflect the change of the transmural gradient,and more research needed to explore its value for clinical application.

AIM:To investigate whether crocin alleviates right ventricular injury induced by monocrotaline(MCT)in rats with pulmonary arterial hypertension(PAH),and to explore the underlying mechanisms.METHODS:Forty male SD rats were randomly divided into 4 groups:normal group,PAH group,crocin group and sildenafil group,with 10 rats in each group.The rats in PAH,crocin and sildenafil groups received subcutaneous injection of MCT(50 mg/kg)to establish the PAH model.Starting from the day of MCT injection,the rats in crocin group received crocin(200 mg/kg),the rats in sildenafil group received sildenafil(30 mg/kg),and those in PAH and normal groups were orally gavaged with an equal volume of saline once daily.After 4 weeks,measurements of right ventricular systolic pressure(RVSP),mean pulmonary artery pressure(mPAP),right ventricular hypertrophy index(RVHI)and right ventricular mass index(RVMI)were taken for the rats in each group.Tissue staining was conducted to observe pathological changes in the right ventricle,and the expression levels of inflammatory factors(IL-1β,IL-6 and TNF-α),the p38 MAPK/NF-κB inflammato-ry pathway,CCL2,CCR2,and the macrophage marker CD68 were assessed.RESULTS:Compared with PAH group,the rats in crocin and sildenafil groups exhibited significant reductions in RVSP,mPAP,RVHI and RVMI(P＜0.05).Right ventricular tissue displayed no evident infiltration of inflammatory cells or proliferation of collagen fibers.The down-regulation of the p38 MAPK/NF-κB pathway and inflammatory factors(IL-1β,IL-6 and TNF-α)was significant(P＜0.05).Additionally,the CCL2/CCR2 pathway and the infiltration of CD68+ macrophages were markedly decreased(P＜0.05).CONCLUSION:Crocin effectively mitigates right ventricular damage in MCT-induced PAH rats,with its effica-cy comparable to that of sildenafil at the dosage utilized in this experiment.Some protective mechanisms of crocin may be attributed to its regulatory effects on inflammation.