BACKGROUND:Previous in vitro studies mainly focused on morphological and nerve marker aspects in the study of bone marrow mesenchymal stem cells (BMSCs)-derived neuron-like cells,but less focused on the olectrophysiological properties of neuron-like cells following differentiation.OBJECTIVE:To study the electrophysiological changes of differentiation from BMSCs into neuron-like cells after induction by brain-derived neurotrophic factor (BDNF)/basic fibroblast growth factor (bFGF)/alltrans-retinolc acid (AT-RA).DESIGN,TIME AND SETTING:The cytological comparative in vitro study was performed at the Department of Neural Cell Laboratory,Tianjin Huanhu Hospital and College of life Science of Nankai University from June 2005 to October 2007.MATERIALS:Totally 3 male Wistar rats (6-week old,weighing about 160 g) were used in this study.METHODS:BMSCs were cultured and purified by their characteristic of plastic adhesion in vitro,and then induced by BDNF,bFGF and AT-RA,and differentiate into neuron-like cells.Whole-cell patch damp technique was used to detect cell membrane current prior to and 3 days following induction.MAIN OUTCOME MEASURES:MSC phenotype was determined by flow cytometry.Cell morphology was observed under the inverted microscope before and after differentiation.Neuron specific enolase expression was assessed by immunocytochemistry.Whole-cell current results were measured.RESULTS:Flow cytometry results demonstrated that CD90 positive rate (99±3)%,CD31 (3.4±0.8)%,and CD34 (0.3±0.1)%.This indicated that these cells were undifferentiated stem cells,with purity of 95%.Undifferentiated MSCs under the optical microscope were mostly fiat cells with processes,similar to fibroblast-like cells.Neuron-like cells appeared 3 days following induction.Immunocytochemistry results showed that MSCs before induction were weakly positive for neuron specific enolase,but strongly positive for neuron specific enolase.At 72 hours,the differentiated rate was (24.01±3.76)%.The peak currents of outward currents in neuron-like cells were significantly higher in the induction group compared with the control group (P ＜ 0.05),but no inward Na~+ current was detected.CONCLUSION:(bFGF & BDNF)/AT-RA could induce the differentiation of MSCs into neuron-like cells.These cells showed the tendency to differentiate into mature neurons,though having no electrophysiological properties of mature neurons.

Objective To discuss the feature of transient intrahepatic cholestasis in early stage of orthotopic liver transplantation. Methods Based on the review of early (within 1 month) postoperative hyperbilirubinemia in consecutive 200 patients undergoing liver transplantation, we summarized the occurrence, development and outcome of early postoperative intrahepatic cholestasis. Results Early transient intrahepatic cholestasis was identified in 112 patients. The characteristic of early intrahepatic cholestasis is that DBIL and?-GT increasingly elevated from the second or third day postoperatively, with a peak on the 7 - 14th d, then descended to normal level on approximately 21 -28th day. The average peak level of DBIL and?-GT were( 157. 32?82. 08)?mol/L and (172?80) IU/L respectively. During the period of DBIL and?-GT ascending, AST and ALT kept descending, and within 1 week it could fall to normal level. Acute rejection, drug toxicosis and bile duct obstruction were excluded. Conclusions Postoperative early transient intrahepatic cholestasis associated with ischemia-reperfusion injury has its special clinical process and most patients recover themselves without the need for special therapy.

Objective To study the effects of restructuring tissue inhibitor of matrix metatloproteinase-3 (rhTIMP-3) in combination with cisplatin (DDP) on the growth and apoptosis of A549 lung cancer cell line.Methods We made individual and combined use of different concentrations of rhTIMP-3 and DDP on A549 cells.Methyl thiazoyl terazolium (MTT) colorimetry was used to analyze cell growth inhibition,and flow cytometry technique was used to determine the cell cycle distribution and apoptosis rate.Results rhTIMP-3 and DDP both could inhibit the proliferation of A549 cells.rhTIMP-3 exerted its effect in the time-and concentration-dependent manners while DDP did so in the concentration-dependent manner;both induced the apoptosis of A549 cells.rhTIMP-3 could make the cells stay in S and G2/M phases,and DDP made the cells stay in S phase.The combination of them obviously strengthened the inhibition of A549 cell growth,and had obvious synergy in inducing apoptosis.Conclusion Both rhTIMP-3 and DDP can inhibit the proliferation of A549 cells and induce their apoptosis.The combined use of them not only can increase the inhibition of cell growth but also has synergy in inducing cell apoptosis.

Objective To study the effect of feto-matemal microchimerism in the treatment of activated human leukocyte antigen (HLA) haploidentical mobilized peripheral blood cells against solid tumors. Methods Genomic DNA samples of 25 pairs of HLA haploidentical donors and recipients were extracted. The donor-derived HLA-DRB loci were detected with nested PCR-sequence specific primer(SSP) typing. The mixed lymphocyte proliferation action between the patients and respective donors, the engraftment of donor's cells and the serum levels of Th1/Th2 type of cytokines were measured with MTT,FISH and EIJSA method respectively. The survival time of patients with or without feto-matemal microchimerism were compared as well. Results Using nested PCR-SSP typing, the positive rates of feto-maternal microchimerism in the 25 pairs of HLA haploidentical donors and recipients were 40% in the maternal/children pairs and 0 in the paternal/children pairs. The chimerism positive patients showed less proliferation activity when cocultured with respective donors as compared with unrelated ones (P=0.03).Only one chimerism positive patient experienced the engraft of donor's cell 3 months after treatment as the donor derived XX chromosome was identified with FISH. When the data of chimerism positive patients were deleted, the serum levels of IFNγ 1 month after treatment dropped dramatically from 171.4 (26. 3～258.4) ng/L to 29. 4(1.2～39.9)ng/L. The survival time in chimerism positive patients of the maternal/children pairs was significantly longer than that in chimerism negative patients, which was (31.2±4. 3) months and (11.1±3.3) months, respectively (P=0.036). Conclusion Feto-maternal microchimerism might induce anergy in the HLA haploidentical donors, favor the engraftment of donor's progenitors and maintenance of positive microenvironment and prolong the survival time.

Objective:To investigate the efficacy of double balloon enteroscopy (DBE) in the treatment of bleeding from small intestinal vascular lesion and risk factors of bleeding recurrence .Methods:From April 2013 to May 2020, at Air Force Medical Center, the clinical data of 65 patients with confirmed or suspected bleeding from small intestinal vascular lesion were retrospectively analyzed. The patients were divided into DBE treatment group (patients of Yano classification 1a and 1b received argon plasma coagulation, and patients of Yano classification 2 and 3 accepted combination of titanium clip and submucosal injection of lauromacrogol sclerosing agent) and non-DBE treatment group (traditional treatments such as stopping anticoagulant or antiplatelet drugs, blood transfusion, and iron supplementation). The bleeding recurrence of patients with single small intestinal vascular lesion between DBE treatment group and non-DBE treatment group, and patients with single or mulitiple vascular lesion of DBE treatment group were compared. Univariate analysis was used to analyze the clinical data of patients with or without recurrent bleeding. Multivariate logistic regression model was used to analyze the independent risk factors and protective factors of recurrent bleeding in small intestinal vascular lesion. Independent sample t test, chi-square test and Fisher exact probability method were used for statistical analysis. Results:Forty-four (25 of single vascular lesion and 19 of multiple vascular lesion) patients were diagnosed with small intestinal vascular lesions and received DBE treatment (DBE treatment group). Twenty-one patients with single vascular lesion accepted traditional treatment (non-DBE treatment group). The recurrent rate of bleeding in patients with single vascular lesion of DBE treatment group was lower than that in patients with single vascular lesion of non-DBE treatment group and patients with multiple vascular lesion of DBE treatment group (24.0%, 6/25 vs. 71.4%, 15/21 and 12/19), and the differences were statistically significant ( χ2=10.348 and 6.848, P=0.001 and 0.009). The results of univariate analysis showed that the proportion of blood transfusion, hypertension, complicated with valvular heart disease and DBE treatment in patients with rebleeding or not rebleeding from small intestinal vascular lesion was different with statistically significant (69.7%(23/33) vs. 37.5%(12/32), 51.5%(17/33) vs. 18.8%(6/32), 42.4%(14/33) vs. 12.5%(4/32) and 54.5%(18/33) vs. 81.2%(26/32), χ2=6.777, 7.628, 7.265, and 5.298, all P<0.05). The results of multivariate logistic regression analysis indicated that blood transfusion during the course of disease (odds ratien ( OR)=3.736, 95% confidence interval ( CI) 1.082 to 12.898, P=0.037) and complication with valvular heart disease ( OR=4.916, 95% CI 1.107 to 21.829, P=0.036) were independent risk factors of bleeding recurrence in patients with small intestinal vascular lesions. DBE treatment was the protective factor of bleeding recurrence in patients with small intestinal vascular lesion ( OR=0.214, 95% CI 0.057 to 0.808, P=0.023). Conclusions:DBE is effective in the treatment of small intestinal vascular lesion bleeding, especially for single vascular lesion. Blood transfusion during disease course and complication with valvular heart disease are independent risk factors for bleeding recurrence in patients with small intestinal vascular lesion.

OBJECTIVETo investigate the changes in rat cardiac function and myocardium ultrastructure in response to ouabain treatment.

METHODSTwenty-four male SD rats were randomized into two equal groups to receive daily intraperitoneal injection of ouabain or saline for 4 consecutive weeks, and their systolic blood pressure (SBP) was recorded weekly. After 4 weeks of injection, echocardiography was performed and the hemodynamic parameters were measured by invasive cardiac catheterization, and the changes in myocardium ultrastructure observed using transmission electron microscopy.

RESULTSAfter 4 weeks of ouabain injection, no significant changes in the mean SBP occurred in comparison with the saline group, but echocardiographic examination showed significant increases in the left ventricular end-diastolic and end-systolic diameters, septum thickness, posterior wall thickness, left ventricular mass and isovolumetric relaxation time but significantly lowered E/A ratio, ejection fraction and fractional shortening after ouabain treatment (P<0.05). Invasive monitoring revealed significant attenuation of the left ventricular developed pressure, rate of pressure development (+dp/dt) and rate of pressure decay (-dp/dt), and increment of the left ventricular end diastolic pressure. Myofibrillar fragmentation, swelling of the cardiac myocytes, absence of the Z line, increases of the mitochondria and collagen fibers were found in ouabain group by transmission electron microscopy.

CONCLUSIONOuabain can induce left ventricular enlargement, cardiac wall thickening, myocardial ultrastructural alterations, systolic and diastolic dysfunction in rats before blood pressure elevation is detected, indicating that ouabain can directly cause cardiac damage in rats.

Animals ; Echocardiography ; Heart ; drug effects ; physiopathology ; Male ; Microscopy, Electron, Transmission ; Myocardium ; pathology ; ultrastructure ; Ouabain ; pharmacology ; Random Allocation ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo study the identification method of Pterocephalus hookeri.

METHODThe microscopical, Physicochemical and TLC methods were used.

RESULT AND CONCLUSIONThe convenient and effective identification methods for P. hookeri were established, which provide basis for its quality standard and development.

Chromatography, Thin Layer ; Drugs, Chinese Herbal ; analysis ; Magnoliopsida ; anatomy & histology ; chemistry ; Pharmacognosy ; Plant Leaves ; anatomy & histology ; chemistry ; Plant Roots ; anatomy & histology ; chemistry ; Plants, Medicinal ; anatomy & histology ; chemistry ; Quality Control

OBJECTIVETo investigate the effect of endothelin and its receptors on ouabain-induced hypertension in rats.

METHODSMale Sprague-Dawley (SD) rats were treated with ouabain or saline for 6 weeks and their systolic blood pressure (SBP) were recorded weekly. At the end of 2, 4 and 6 weeks, respectively, the plasma and left ventricle endothelin contents were measured by radio-immunoassay, and real-time quantitative RT-PCR was employed to determine the mRNA level of endothelin type A receptor (ETAR) and type B receptor (ETBR) in the left ventricle, and the protein expressions of ETAR and ETBR were examined by immuno-histochemistry.

RESULTSAfter 4 weeks of intraperitoneal ouabain injection, the mean SBP in ouabain group increased till reaching a level significantly higher than that in the control group after 6 weeks (P<0.001). The plasma and left ventricle endothelin contents were significantly increased after 2 weeks of ouabain injection (P<0.01), and similarly, increased ETAR mRNA was observed. After 4 weeks of treatment, ETAR mRNA was increased continuously and the protein expression of ETAR upregulated in ouabain group as compared with the control group. The transcription and protein expression of ETBR were not altered by ouabain treatment.

CONCLUSIONBefore detectable blood pressure elevation occurs, endothelin concentration and ETAR can be already upregulated in ouabain-induced hypertensive rats, suggesting that endothelin might be involved in the cardiovascular effects of ouabain via an action on ETAR.

Animals ; Endothelins ; biosynthesis ; blood ; genetics ; Hypertension ; chemically induced ; genetics ; metabolism ; Immunohistochemistry ; Male ; Myocardium ; metabolism ; Ouabain ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Receptor, Endothelin A ; genetics ; metabolism ; Receptor, Endothelin B ; genetics ; metabolism ; Receptors, Endothelin ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

Objective To investigate the effects of different irradiation techniques on dose distribution in target volume and normal tissues after the radical surgery for gastroesophageal junction adenocarcinoma,and to provide the optimal regimen for clinical treatment.Methods A total of 9 patients with gastroesophageal junction adenocarcinoma who underwent radical esophagus-proximal gastrectomy or total gastrectomy were enrolled.The therapeutic regimens of five-field static intensity-modulated radiotherapy (IMRT),volumetric-modulated arc therapy (VMAT),and helical tomotherapy (HT) were designed for each patient,and the dose-volume histogram was used to evaluate the effects of different irradiation techniques on the conformity index (CI) and homogeneity index (HI) of target volume and the surrounding normal tissues. The prescribed dose was 45 Gy at 1.8 Gy/fraction.The patients received oral S-1 as concurrent chemotherapy at a dose of 80 mg/(m 2? d) twice a day during radiotherapy.Results Compared with IMRT and VMAT,HT had better CI and HI of the target volume,as well as a better protective effect on the intestinal tract and bone marrow.Compared with IMRT and HT,VMAT had a lower V20 and V30 for the left kidney and a lower V30 for the heart,while IMRT had lower V5 and V10 for both lungs;V20 and mean dose showed no significant differences between the three techniques.HT had the highest mean sub-field hop count,followed by IMRT and VMAT.Conclusions IMRT, VMAT, and HT can meet the clinical requirements,but besides ensuring the best CI and HI of the target volume,HT has a good protective effect on the intestine and spinal cord and can help to reduce the incidence of adverse events in patients.

Objective To reduce the radiation dose to the hematopoietic bone marrow (hBM) and acute hematologic toxicity (HT) in patients with rectal cancer undergoing intensity-modulated radiotherapy (IMRT).Methods The previously untreated patients with rectal cancer were enrolled in a prospective study.Pelvic magnetic resonance imaging ( MRI) was used to determine and delineate the distribution of hBM,and dose limitations were set (V5<95%,V10<90%,V20<80%,V30<65%).The neoadjuvant therapeutic regimen included concurrent IMRT (95% PTV 50 Gy/25 fractions,2 Gy/fractions),oxaliplatin 50 mg/m2 , qw,and capecitabine 1650 mg/m2 ,1 fractions/d (twice a day during radiotherapy).Results A total of 35 patients were enrolled and completed the therapeutic regimen.The incidence of grade 2-4 HT was 31.4%;among these patients, 9 ( 26%) experienced leucopenia, 6 ( 17%) experienced neutropenia, 1 ( 3%) experienced erythropenia,and 1(3%) experienced thrombocytopenia.No patients experienced grade ≥3 anemia.The multivariate logistic linear regression analysis showed that hBM-V5 was significantly correlated with the lowest counts of leukocytes ( P=0.005),neutrophils ( P=0.002),and platelets ( P=0.017).Conclusions The radiation dose to the hBM in the pelvis on MRI is significantly correlated with the incidence and severity of acute HT in patients with rectal cancer undergoing neoadjuvant concurrent chemoradiotherapy.Clinical Trial Registry ClinicalTrials.gov,registration number:NCT01863420.