Objective To identify a detailed biological characterization of mesenchymal stem cells (MSCs) isolated from hu‐man umbilical cord(UC) tissue regarding their morphology ,immunophenotype ,purity and proliferative capacity and establish a rea‐sonably cultured and amplified system .Methods After stripping off arteries and veins ,the remaining parts of umbilical cord were cut into 1 mm3 small sections and cultured with DMEM/F12 containing 10% fetal bovine serum .Adhere cells were obtained and the morphology of the cells was observed under inverted phase contrast microscope .The growth curves of them were drawn by CCK‐8 and the cell cycle and surface antigens (CD29 ,CD73 ,CD90 ,CD105 ,CD31 ,CD14 ,CD34 ,CD45 ,CD11b ,HLA‐DR) were detected by flow cytometry .Results Seven to ten days after primary culture ,adhere cells came out of fragments .The MSCs harvested were a high purity and mainly presented as a fibroblast‐like morphology .UC‐MSCs had a strong ability of proliferation through the cell growth curve .The special surface antigens CD29 ,CD73 ,CD90 ,CD105 were positive expression ,while CD31 ,CD14 ,CD34 ,CD45 , CD11b ,HLA‐DR were negative .More than 80% cells of MSCs were found at G0/G1 phase .Conclusion Human UC‐MSCs could be cultured and proliferated in vitro .

Objective To investigate the perioperative nursing of patients with Stanford type B aortic dissections treated with endovascular graft exclusion.Method The clinical data of 22 cases undergoing thoracic endovascular aortic repair from February 2011 to August 2013 were retrospectively analyzed to summarize the nursing experience.Results Twenty-two patients survived successfully through operation.One case had retrograde type A dissection after operation,another 15 had hyperthermia,and all of them were cured and discharged due to symptomatic treatment.The postoperative 3 months follow-up showed no type I endoleak.Conclusion Preoperative psychological nursing,postoperative blood pressure control,nursing of complications,strengthening instruction of diet and physical activity,are critical for the promotion of early rehabilitation of patients after discharge.

ObjectiveTo explore the effect of JNK-c-Jun signal pathwayon connexin 43 (Cx43) expression and its role in renal tubular epithelial-myofibroblast transition (TEMT) induced by TGF-β1.Methods Normal rat kidney tubular epithelial cells(NRK-52E) were cultured in Dulbecco's modified eagle medium(DMEM) with 10％ fetal bovine serum,then were randomly divided into 3 groups: control group,TGF-β1 group (treated with TGF-β1 10 μg/L),and TGF-β1+SP600125 (selective JNK inhibitor,50 μmol/L) group. The protein expressions of JNK,c-Jun,α-SMA,Cx43 and E-cadherin were assayed by immunocytochemistry and Western blotting.The Cx43mRNA was assayed by RT-PCR.Gap junction intercellular communication(CJIC) was measured by fluorescence recovery after photobleaching assay(FRAP).Results TGF-β1increased the expressions of JNK,c-Jun and α-SMA(P＜0.05),reduced the expressions of Cx43 and E-cadherin (P＜0.05),and inhibited GJIC of NRK-52E(P ＜0.05).SP600125 could alleviate the above expressions changes and enhanced GJIC induced by TGF-β1.Conclusion JNK-c-Jun signal pathway induces TEMT ofNRK-52E treated with TGF-β1 viadown-regulation of connexin 43expression and inhibition of GJIC.

Continuous renal replacement therapy has become an important organ support method for children with liver failure.Anticoagulation is a key step in determining the success of this treatment.Traditional systemic heparin anticoagulation has a high risk of inducing thrombocytopenia and bleeding.Protamine antagonistic heparin local anticoagulation performance is unsatisfactory.Anticoagulation without heparin is difficult to develop smoothly in children, especially low-weight infants.In recent years, regional citrate anticoagulation has gradually received attention as a new type of anticoagulation.This article reviewed the clinical application of this anticoagulation technology.

Aim To generate m3AChR-G11 fusion protein in baculovirus-Sf9 cells and test the couping function,the interation and the influence factors be-tween m3AChR and G11 protein,as well as screen the specific ligands for m3AChR. Methods m3AChR-G11 fused DNA was generated through a two-step PCR and then expressed in Sf9 cells to produce fusion protein. The total concentration for membrane protein was de-tected by BCA method,[3H]QNB and [35 S]GTP?S binding experiment as perfomed to study the function of m3AChR-G11 fusion protein. Results The expression level of m3AChR-G11 was 7. 76 ? 10 -9 mol?g -1. The affinity of GDP to G11 partner changed in the presence of different muscarinic ligands. IC50 values of GDP in the presence of ACh,Pilo,CCh,MCN-A-343,Atro,4-Damp and Dafi were 82. 2,93. 70,12. 10,14. 30, 1. 93,1. 37,0. 72 ? 10 -6 mol ? L -1 respectively,and that in the absence of muscarinic was 1. 99 ? 10 -6 mol ?L-1.Concluslons The m3AChR-G11 fusion protein has the pharmacological specificity of m3 receptor and the efficient coupling interaction of the two partners. Affinity of GDP to ligand-bound fusion protein represents the species of muscarinic ligands. This is helpful in screening and detecting the new specific ligands to muscarinic receptors.

Objective To evaluate the clinical value of transesophageal echocardiography (TEE) on minimally invasive surgical closure of heart septal defects. Methods Thirty-four patients with atrial septal defect (ASD) and 38 patients with ventricular septal defect(VSD) were selected by transthoracic echocardiography (TTE) prepared for minimally invasive surgical closure. TEE was performed to choose appropriate occluder and guide occluder release during the procedure. The immediate closure effect also evaluated by using TEE. A week follow-up was done by using TTE. Results Successful occlusion was in 32 patients with ASD and 29 patients with VSD. On 1 week follow-up,neither displacement for the occluders nor residual shunt except minimal residual shunt in 3 patients. The ventrieular remodeling was improved, the valvular regurgitation and pulmonary arterial pressure decreased. Conclusions TEE is important in minimally invasive surgical closure of atrial, ventricular septal defects, especially in choosing the candidate for the procedure and occluder, guiding occluder released and evaluating the procedure.

Objective Using M2-Gi1α fusion protein expressed by baculovirus-Sf9 cell system to find the specific ligand for M2 receptor and detect the interaction of the two parts of the fusion protein.Methods The fused M2-Gi1α cDNAs were generated in a two-step PCR and then expressed in Sf9 cells.[3H]QNB and[35S]GTPγS binding experiments were employed to study the function of M2-Gi1α fusion protein.Results The expression level of M2-Gi1α fusion protein was 8.44±0.39 nmol·g-1 protein.The affinity of GDP to the Gi1α part changed under the affection of different ligands.The IC50 value in the appearance of acetylcholine,oxotremorine,arecoline,atropine,fangchinoline,levitimide were 21.35 μmol·L-1,23.86 μmol·L-1,11.91 μmol·L-1,0.13 μmol·L-1,1.05 μmol·L-1,1.75 μmol·L-1,and 2.5 μmol·L-1 when there was no ligand.Conclusion The M2-Gi1α fusion protein expressed in baculovirus-Sf9 cell system has pharmacological specificity for M2 receptor and the efficient coupling function between the two parts.The M2-Gi1αfusion protein is a helpful tool for detecting the new specific ligands of the M2 receptor.

AIM: To selecte the best process for Kangfuxin Drop Pills. METHODS: With appearance,weight coefficient of variation and dissolution time for index sign,orthogonal design was applied. RESULTS: The results indicated that the excellent factors and levels were as follows: the scale between PEG6000 and drug extraction was four to one,the temperature of drug fluid was at 85 ?C,drip rate was 40 drops/min. CONCLUSION: The method adopted is steady and feasible.

Objective To investigate the clinical significance of CD55,CD59 and Aeromonas hydrophila toxin variant (FLAER) in the diagnosis of anemia and paroxysmal nocturnal hemoglobinuria (PNH).Methods Collected 30 healthy controls,22 cases of PNH,33 cases of aplastic anemia (AA),37 cases of iron deficiency anemia (IDA),45 cases of megaloblastic anemia (MA),30 cases of hemolytic anemia (HA) and 31 cases of myelodysplastic syndrome (MDS) from January 2009 to March 2017,CD55,CD59 and FLAER negative cell ratio of peripheral blood neutrophil of them were detected by multipa rameter flow cytometry.Results The detection rates of FLAER in PNH,AA and MDS groups were higher than those of CD55 and CD59,but there was significant difference in AA (x2 =7.759,5.518,P=0.005,0.019＜0.05).The average CD55,CD59 and FLAER deletion rate in PNH and AA group were significantly higher than those in normal control group and other groups (t=2.163～17.890,P=0.000～0.038＜0.05).The number of FLAER in PNH group was higher than CD59 and CD59 was higher than CD55 with the statistically significant difference (t=2.503 ～ 6.308,P=0.000 ～0.016＜ 0.05).Conclusion CD55,CD59 and FLAER have important value in the diagnosis of PNH and differential diagnosis with other anemia diseases,and can also be used to detect the presence of MDS and AA in patients with PNH.FLAER outperforms CD59,CD59 outperforms CD55.

Aim To study the protective effect of the compound flavones on chronic alcohol induced testicu-lar damage in mice. Methods One hundred SPF C57 BL/6 mice were randomly divided into 5 groups:normal control group, chronic alcohol group, alcohol+low-dose drug group, alcohol+high dose drug group, and drug control group. Chronic alcohol testicular inju-ry model in mice was established by intragastric admin-istration of increasing dose of alcohol every four weeks for 6 months, meanwhile compound flavone interven-tion was in process. The activity of super oxide dis-mutase( SOD) , the levels of malondialdehyde( MDA) and testosterone in testicular tissue were measured. HE staining was used to observe testicular histomorpholo-gy, and ultrastructure changes were detected by elec-tron microscope. Results In chronic alcohol group, MDA content was obviously increased, while SOD and testosterone levels were decreased compared with nor-mal control group ( P <0. 05 ) . In addition, germinal epithelium, support cells and sperm production at all levels showed degraded degeneration in chronic alcohol group. However, compound flavonoids could success-fully reverse alcohol-induced testicular damage in a dose-dependent way. Conclusion The compound of flavones may have therapeutic potential for alcohol-in-duced testicular injury through inhibiting lipid peroxi-dation and increasing the level of testosterone.