Objective To study the clinical efficacy of human blood albumin and small dose heparin in treatment of severe preeclampsia and its effect on the expression of Endoglin,vascular endothelial growth factor (VEGF)and its receptors (Flt)in placental tissue.Methods 60 cases of severe preeclampsia collected in weifang yidu central hospital from October 2012 to January 2014 were randomly divided into experimental group and control group,each had 30 cases.Control group were given oral antihypertensive,expansion and diuretic treatment.Experiment group were received injection with 500 mL 5% glucose,60 mL 25% magnesium sulfate and 25 mg heparin on the basis of control group,one time each day,for successive seven days.Another 30 cases of normal full-term pregnancy for cesarean section delivery collected in our hospital during the same period were as blank group.Mean arterial pressure (Kpa)and 24 h urine protein quantitative and clinical efficacy were observed and compared between experimental group and control group before and after treatment.curative effect. Microscopically observed treatment after Endoglin in blank group, control group and experimental group placenta tissue,The expression of Endoglin,VEGF and Flt 1 in placental tissue among three groups were observed by microscope, and their positive expression rate were compared. Results The mean arterial pressure and urine protein were all improved after treatment in control group,and experimental group,but the latter was more obvious than the former(P<0.05 ).The expression location of Endoglin,VEGF and PLT-1 in three groups were the same with microscopic observation.Compared with blank group,the positive expression rate of Endoglin,VEGF protein and PLT-1 in placenta of other two groups were higher(P<0.05),and those in experimental group were more higher than control group(P<0.05).Conclusion Human blood albu min and small dose heparin can effectively improve the indexes of quantitative Kpa and 24 h urine protein in severe preeclampsia, reduce the expression of Endoglin,VEGF and Flt 1 protein in placental tissue.

Objective To investigate the risk factors of postoperative complications of delayed hemorrhage,perforation and digestive tract stenosis after endoscopic submucosal dissection (ESD).Methods The complete data of 793 patients with digestive tract disease who underwent the endoscopic submucosal dissection in the Department of Digestive Surgery in our hospital from January 2011 to December 2014 were retrospectively analyzed.All of the patients were divided into delayed hemorrhage group (n =67) and nonbleeding group (n =726);perforation group (n =47) and non-perforation group (n =746);and digestive tract stenosis group (n =38) and non-stenosis group (n =755).The clinical basic data,lesion related data,and operation related data were independent risk factor and analyzed by single factor analysis and Logistic multiple factor regression analysis.Results The incidence of delayed bleeding,perforation and stenosis in patients with ESD were 8.45％,5.93％,and 4.79％,respectively.The results of single factor analysis:the risk factors for delayed bleeding were long-term use of anticoagulant drugs,gastric sinus disease,lesion diameter,and lesion excision (P ＜ 0.05).The risk factors for postoperative perforation were the diameter of the lesion and the time of operation (P ＜ 0.05).The risk factors of digestive tract stenosis were the esophageal lesions,the diameter of the lesion,and the depth of the lesion to the intrinsic muscle layer (P ＜ 0.05).The results of multi factor Logistic regression analysis:the risk ranking of risk factors for delayed bleeding was gastric antrum occurrence lesion ＞ lesions graded resection ＞ long-term use of anticoagulants ＞ lesion diameter (≥5 mm).The risk ranking of risk factors for perforation was operation time (≥90 mm) ＞ lesion diameter (≥5 mm).The risk ranking of risk factors for digestive tract stenosis was esophageal lesion ＞ lesion diameter (≥ 5 mm) ＞ lesions depth to the muscularis propria.Conclusions For long-term anticoagulation,gastric antrum and fractional resection lesions of patients should pay attention to delayed bleeding.Patients with long operation time are easy to cause postoperative perforation.For long-term anticoagulation,gastric antrum and fractional resection lesions of patients should pay attention to delayed bleeding.

BACKGROUND:Currently,there is not a standard method for in vitro culture and large scale amplification of umbilical cord blood mesenchymal stem cells(UCB-MSCs).OBJECTIVE:To investigate the isolation,purification and culture of UCB-MSCs in vitro,and to detect its surface marker variation.METHODS:The monocytes were harvested from UCB using 1.077 g/cm3 lymphocytes separating solution and density gradient centrifugation,followed by incubation in an incubator containing 5%CO2 at 37℃.The cell morphological changes were observed at different time points and the expression of surface marker was detected using flow cytometry.RESULTS AND CONCLUSION:The monocytes isolated from the UCB grew initially into numerous hematopoietic cell clones,most of which were granulocyte/macrophage colony-forming units and burst forming unit-erithroid,increasing by(37.1±2.3)and (10.4±1.7),respectively.Switzerland staining showed most of them were granulocyte clones(80,1±85.2)%,next was erythroid clones(14.2±1.8)%.At 7 days after culture,some shuttle fibroblast-like cells and fiat osteogenic-like cell spread the whole plastic well.At 14 days after culture,flow cytometry showed CD38+ cells accounted for 1.64%,and CD34+/CD38+ cells accounted for 1,71%,and CD34+/CD38- were 0.55%.PI+ and Annexin-V+ cells accounted for 0.05% and 0.18% respectively.At 21 days after culture,CD38+,CD34+/CD38+ and CD34+/CD38- cells were 74.32%,1.61%,and 0.24%.The results reveled that UCB-MSCs can be isolated and cultured in vitro.

Objective To investigate the blood pressure circadian rhythm in patients with IgA nephropathy by ambulatory blood pressure monitoring and explore its role in the disease progression. Methods A cross sectional study was carried out.Blood pressure rhythm was studied by ambulatory 24-hour monitoring with a portable oscillometric recorder in selected patients with primary IgA nephropathy.The term dipper was described as blood pressure during night dropped at least 10％ below daytime blood pressure.The term non-dipper referred to those in whom the nocturnal decline in blood pressure was less than 10％.Clinicopathological indices between dipper and non-dipper groups were compared. Results Ninety-three patients completed ambulatory blood pressure monitoring among whom 68 (73％) patients were non-dipper.The frequency of non-dipper was 70％,70％ and 81％ in the patients at chronic kidney disease stage 1,2 and 3 or more.The frequency did not differ among these three group patients (P=-0.587).77％ of patients with hypertension and 69％ of patients with normotension were non-dipper (P=0.373).The disappearance of blood pressure circadian rhythm in IgA nephropathy was not influenced by age,gender,blood pressure,proteinuria,renal function and renal pathology lesions.Among the patients who were followed up regularly for more than 12 months (n=54),patients in the dipper group had a trend of slower eGFR decline rate than those in non-dipper group albeit the difference was not significant (P=0.329).Subgroup analysis revealed that in patients with hypertension and non-dipper (n=29),the eGFR decline rate was much faster than that in dipper group[(-6.79±11.58 )vs (-0.34±1.74) ml ·min-1 ·(1.73 m2)-1·year-1,P=0.019]. Conclusions Most patients with IgA nephropathy present disappearance of blood pressure circadian rhythm,even among those at an early stage or without hypertension.The loss of blood pressure rhythm may be associated with a rapid renal function decline rate in those with hypertension.

BACKGROUND: Stem cells are relatively primitive cells possessing the capabilities of self-renewal, high proliferation and multi-potential differentiation in vivo under certain conditions. Pancreatic stem cells and umbilical cord blood mesenchymal stem cells (MSCs) may serve therapeutic purpose clinically, but they are still difficult to culture in vitro at present.OBJECTIVE: To explore the method for isolation, purification and culture of pancreatic stem cells and umbilical cord blood MSCs in vitro and observe their morphological changes during culture in vitro.DESIGN: Completely randomized experiment with repeated measurement.SETTING: Stem Cell Research Center, Teaching and Research Division of Physiology, Medical School of Zhengzhou University.MATERIALS: This experiment was conducted in the Stem Cell Research Center, Teaching and Research Division of Physiology, Medical College of Zhengzhou University, between April 2004 and January 2005. Ten to fifteen newborn SD rats (1-3 days) were selected for culture in vitro of pancreatic stem cells, and fresh umbilical cord blood was collected from healthy woman (24-35 years old, with informed consent) at full-term delivery for culture in vitro of umbilical blood SMCs.METHODS: The abdomen of the newborn SD rat was opened under aseptic condition to obtain the pancreas, which was cut into small tissue blocks and digested with type-V collagenase for islet isolation. The isolated islets were purified in continuous roller-bottle culture. Umbilical cord blood was freshly collected for isolating the monocytes by means of density gradient centrifugation in lymphocyte separation medium (with density of 1.077 g/cm3). The islet cells and umbilical cord blood monocytes were cultured in the incubator at 37 ℃ with 5% CO2. The morphological changes of the cells were observed at designed time points and flow cytometry was used to determine the expression of cell surface molecules.MAIN OUTCOME MEASURES: The isolation and culture of pancreatic stem cells and umbilical cord blood MSCs, and their morphological changes during culture in vitro.RESULTS: During culture in vitro, the fusiform islet progenitor cells showed adherent polar growth and continuous proliferation, which covered the whole bottom of the flask after 12-14 days and could be subcultured for passages. However round cells appeared after removal of the growth factor and serum in the culture medium. The monocytes isolated from the umbilical cord blood grew initially into numerous hematopoietic cell clones, most of which proved to be granulocyte clones by Switzerland staining. Seven days later, flat flask wall-adhering epithelial cells and long fusiform fibroblasts were observed mixed with a number of osteoclasts. As the cell culture was prolonged, the cell number increased steadily.CONCLUSION: Pancreatic stem cells and umbilical cord blood SMCs can be cultured in vitro for further experiments.

ObjectiveTo investigate the clinical significance and histological origin of glomerular epithelial proliferative lesion in patients with focal segmental glomerulosclerosis (FSGS). MethodsSeventy-four patients with idiopathic FSGS hospitalized in Peking University First Hospital from Jan. 2000 to Dec.2005 were enrolled in this study. Patients were classified into two groups according to with or without glomerular epithelial proliferative lesion. Estimation of active and chronic pathological scores was carried out using a semi-quantitative grade system by two pathologists. Clinical and pathological characteristics were compared between two groups. Immunohistochemical studies were performed to analyze the histological origin of glomerular epithelial proliferative lesion. ResultsThirty-one patients with glomerular epithelial proliferative lesion showed shorter interval from presentation to biopsy (P<0.05), higher percentage of nephrotic syndrome (NS) (P<0.05), higher frequency of segmental glomerulosclerosis(P<0.05), higher pathological active scores (P<0.05) and lower pathological chronic scores (P<0.05)as compared to 43 patients without glomerular epithelial proliferative lesion. Twenty-nine patients were followed up and renal survival rate in patients with glomerular epithelial proliferative lesion (39.7%) was significantly lower than that in patients without glomerular epithelial proliferative lesion (83.3%) (P=0.049). The frequency of glomerular epithelial proliferative lesion and the serum creatinine (Scr) level at biopsy were independent predictors of ESRD (OR value was 1.204, 1.008 respectively ). Glomerular epithelial proliferative lesion did not express mature podocyte markers including WT-1 and pedocalyxin, but stained positive for PCNA, PAX-2 and CK-8. ConclusionsGlomerular epithelial proliferative lesion represents the pathological change of acute stage and active lesion of FSGS, and also may be the pathological marker of severe clinical presentation and worse renal survival. Glomerular epithelial proliferative lesion may be derived from proliferation of parietal epithelial proliferation or de-differentiated podocytes.

Objective To develop a rapid,sensitive,specific and accurate quantitative duplex real-time PCR assay for detection of Listeria monocytogenes and Shigella.Methods Two sets of specific primers and probes were selected according to Listeria monocytogencs hly gene and Shigella ipaH gene.The target hly and iPaH fragments were amplified by PCR,and used to construct recombinant pGEM-T-hly and pGEM-T-ipaH respectively.The two recombinant circular plasmid DNAs were linearized with EcoR I that did not cut within the target DNA fragment.The ten-fold dilutions of plasmid were subjected to the standard quantitation curve in duplex real-time PCR assay.Various genomic DNAs of Listeria innocua,Listeria weshimeri,Salmonella,Staphylococcus aureus,Bacillus subtilis,Escherichia coli and Proteus were used as negative controls to confirm the specificity of duplex real-time PCR assay.The assay was also used to detect Listeria monocytogenes and Shigella in artificially contaminated sterilized skim milk.Results The recombinant plasmids were constructed successfully,hly probe(rAM and TAMRA double labelled)and ipaH probe (HEX and TAMRA double labelled)were used to develop an optimized PCR successfuliv.Conclusion The selected primers and probes showed high specificity for these two target bacteria,the linear range of the assay was good(105-101 copies/μl,R2≥0.998)and sensitivity Was 10 copies/PCR.Following a DNA extraction method which combined EZ Spin Colum Genomic DNA Isolation Kit(BBI)/Phenol-chloroform,the sensitivity of assay Was 102CFU/ml for both Listeria monocytogenes and Shigella in artificially contaminated sterilized skim milk,which equivalents to 10 CFU/PCR.

Objective To explore the inhibitory effects of lobaplatin and cisplatin and their regulation of apoptosis-related genes in ovarian cancer cells in nude mice.Methods SKOV3 cells were implanted into nude mice.In monotherapy treatment study,the nude mice bearing human SKOV3 cells were randomly divided into control,lobaplatin,and cisplatin groups,with 7 mice in each group.The mice in each group were received corresponding treatment.The volume of tumor and the weight of nude mice were measured three times per week,respectively.Tumor inhibitory rate was calculated.The protein expressions of bax and bcl-2 were detected by flow cytometry.Results The growth inhibitory rate was 47.2％ in lobaplatin group and 42.8％ in cisplatin group,without significant difference between two groups (P ＞ 0.05).The expression of bcl-2 was decreased but the bax was increased in lobaplatin and ciaplatin groups compared to the control group.Conclusions Lobaplatin can significantly inhibit the growth of ovarian cancer cells,induce apoptosis by down-regulation of bcl-2 and up-regulation of bax.

Objective To realize antimicrobial resistance and carrying status of OXA carbapenemase among imi-penem-resistant Acinetobacter baumannii (IRAB)isolated from patients of Hohhot,so as to provide guidance for the prevention and control of healthcare-associated infection(HAI)caused by multidrug-resistant Acinetobacter bauman-nii .Methods 49 IRAB isolates from 3 tertiary first-class hospitals in Hohhot between January and December 2012 were collected,antimicrobial susceptibility testing was performed by Kirby-Bauer disk diffusion method,four geno-types(blaOXA-51-like ,blaOXA-23-like ,blaOXA-24-like ,blaOXA-58-like )of OXA carbapenemase were detected by polymerase chain reaction (PCR).Results All 49 isolated IRAB strains were found to be highly resistant to antimicrobial agents (81 .63%-100.00%)except to minocycline (8.16%);blaOXA-51-like was identified in 49 strains (100.00%),42 (85.71 %)of which also carried blaOXA-23-like gene ,blaOXA-23-lik and blaOXA-51-like were both found in three hospital, blaOXA-24-like and blaOXA-58-like weren’t found.Conclusion IRAB strains present multidrug resistance,resistant to mi-nocycline is the lowest;blaOXA-23-like is the main drug-resistance mechanism of IRAB in Hohhot.