OBJECTIVE: To investigate the effect of compound pattern of ceramic bovine bone (CBB) and hydrogel(HG) on attachment, proliferation and differentiation of bone marrow stromal cell (MSC), and to find out the best way of constructing tissue engineered bone. METHODS: CBB, HG and MSC was compounded in different patterns and sequences to form CBB/HG/MSC (group A), HG/MSC/CBB (group B), CBB/MSC/HA (group C) and CBB/MSC (control group). Attachment and morphology of MSC were observed by scanning electronic microscope; the proliferation of MSC was evaluated by cell count; alkaline phosphatase(ALP) activity was examined by histochemistry and type I collagen synthesis was examined by immunohistochemistry staining 5 and 10 days later. RESULTS: In group A, MSC spread better, and ALP activity of group A was significantly higher than that of group B and control group(P lt; 0.01); but there was no significant difference between group A and group C(P gt; 0.05). There was no significant difference in type I collagen synthesis between four groups on the 5th day; but mean gray scale of type I collagen in group B was significantly higher than that in the other groups on the 10th day(P lt; 0.01). CONCLUSION: Different compound patterns of CBB, HG and MSC affect attachment, proliferation, differentiation of MSC. The compound pattern of CBB/HG/MSC is better than the others.
Objective To observe the heterotopic osteogenes is of the autogenou s marrow stromal cells (MSCs) on the ceramic bovine bone(CBB)/hydrogel scaffold (HG) and t he effects of the recombinant human bone morphogenetic protein2 (rhBMP-2) and the transforming growth factor β (TGF-β) on osteogenesis. Methods The auto genous marrow stromal cells were cultured by the mineralized condition medium (1 0%FBS, dexamethasone 10 nmol, L-vitamin C 50 mg/L, βsodium glycerophosph ate D MEM culture medium 10 mmol). At 5 days, the MSCs differentiation was observed b y TypeⅠcollagen, the Mend calcium-cobalt staining, and the Von-Kossa staining. The cell suspension of 5×106/ml was obtained. There were three groups: Group A: added in rhBMP-2（10 μg）TGF-β（0.05 μg）;Group B: added in TGF-β(0.05 μg); and Group C (the control group): without the growth factor. Then, the MSCs loading on CBB/HG were embedded in the autogenous subcutaneous area at 4 and 8 weeks, and the osteogenesis was observed by the HE staining and the modified Mallory’s trichrome staining, with an image analysis. TypeⅠcollagen and the bone m orphogenetic synthesis were examined by the immunohistochemistry stains. Results Most MSCs induced by the mineralized condition medium at 5 da ys became smalle r and polygon-shaped, and the cytodendrite became shorter. The MSCs were observ e d by the Mend calciumcobalt staining. Some brown and black grains were found in the cytochylema. The MSCs were positive for the TypeⅠcollagen immunohistochemi stry stains. At 20 days, the mineralized nubs were found by the Von Kossas stain s. At 4 weeks, some strips of the new bone were observed by the HE staining an d the modified Mallory’s trichrome staining in all the groups. The bone matrix a rea was significantly larger in Group A than in Group B（P＜0.01). The av erag e gray degrees of TypeⅠcollagen were lower in Groups A and B than in Group C. However, there was no significant difference in the bone morphogenesis among the three groups. At 8 weeks, there- were significantly more snatchy strips and macula mature bone formation in Groups A and B than in Group C. The Type Ⅰcollage n and the bone morphogenesis were not significantly different among the three groups. Conclusion The autogenous marrow stromal cells on the ce ramic bovine bon e /hydrogel scaffold can promote the heterotopic osteogenesis, and the combined use of rhBMP-2 and TGF-β is better than the only use of rhBMP-2 or TGF-β i n promoting osteogenesis.
Objective To investigate the effects of ectomesenchymalstem cells on hematopoiesis after total body irradiation in rats. Methods The primary ectomesenchymal stem cells were isolated from E11.5 SD fetal mandibular processes by 25g/L trypsin and cultured with DMEM/F12. The morphology and growthrate were observed by inverted microscope. Eighty SD male rats randomly dividedinto ectomesenchymal stem cells group (n=20), fibroblast group(n=20), saline group(n=20) and control group(n=20), the first three groups were irradiated with 60Co γ rays at 6.0 Gy. The number of their bone marrow nucleated cells was counted after 4 weeks; the forming ability of colony-forming unit-granulocyte macrophage(CFU-GM) and histopathology of bone marrow were also observed. Results The cultured cells displayed monolayer growth and fibroblast-like with 2-4 processes. The ectomesenchymal stem cells could increase the number of bone marrow nucleated cells and peripheral blood white cell count, and improve the forming ability of CFU-GM. After 4 weeks of transplantation, the number of the peripheral blood white cells in group A was more than that in groups B and C(Plt;0.05), the contents of Hb in groups A and D was significantly higher than those in groups B and C(Plt;0.0). After 4 weeks, the bone morrow nucleated cells in group A were significant more than those in groups B and C(Plt;001); CFU-GM in groups A and D was higher than that in groups B and C(Plt;0.01). Conclusion Ectomesenchymal stem cells have characteristics of stem cells. It may improve hematopoiesis recovery of irradiated rats.
Objective To introduce the development of dextran-based hydrogel and its drug delivery system in drug sustained and/or controlled release, and to investigate their application in tissue engineering.Methods Related literature was extensively reviewed and comprehensively analyzed. Results In recent years, great progress was made in the studies of dextran-based hydrogels and study on dextran-based intelligent materials became an investigative hotspot especially in tissue engineering. Conclusion Dextran based hydrogel is considered to be a good potential material in field of drug delivery and tissue engineering. Endowed with new characteristics, a series of intelligent biomaterials can be derived from dextran-based hydrogels, which can be widely used in biomedicine. Further study should be done on the industrialization of its interrelated production.
OBJECTIVE: To investigate the selection and identification of human keratinocyte stem cells(KSC) in vitro. METHODS: According to the characteristics of KSC which can adhere to extracellular matrix very fast, we selected 3 groups of different time(5 minutes, 20 minutes and 60 minutes) and unselected as control group. And the cells were identified by monoclone antibody of beta 1-integrin and cytokeratin 19 (Ck19), then the image analysis was done. Furthermore we analyzed the cultured cells with flow cytometer(FCM) and observed the ultrastructure of the cell by transmission electron microscope(TEM). RESULTS: The cell clones formed in all groups after 10 to 14 days, while the cells of 5 minute group grew more slowly than those of the other groups, however, the clones of this group were bigger. The expression of beta 1-integrin and Ck19 were found in all groups. The positive rate of beta 1-integrin was significant difference between 5 minute group and the other groups (P lt; 0.05). And the expression of Ck19 was no significant difference between 5 minute group and 20 minute group(P gt; 0.05), and between 60 minute group and control group. But significant difference was observed between the former and the later groups(P lt; 0.05). The result of FCM showed that most cells of the 5 minute group lied in G1 period of cell cycle, which was different from those of the other groups. At the same time, the cells of 5 minute group were smaller and contained fewer organelles than those of the other groups. CONCLUSION: The above results demonstrate that the cells of 5 minute group have a slow cell cycle, characteristics of immaturity, and behaving like clonogenic cells in vitro. The cells have the general anticipated properties for KSC. So the KSC can be selected by rapid attachment to extracellular matrix and identified by monoclone antibody of beta 1-integrin and Ck19.
ObjectiveTo assess the method and the results of tricuspid annuloplasty performed(TVP) with the Edwards MC3 ring. MethodsWe retrospectively analyzed the clinical data of 312 patients with functional tricuspid regurgitation(FTR) secondary to left-sided valve disease in our hospital from June 2012 through May 2014. There were 147 males and 165 females at mean age of 55.7±7.3 years. ResultsThere was no death in the patients because of the planting of MC3 ring. The mean follow-up rate was 99.4%(310/312) for 2 patients immigration abroad. The follow-up time was 0-24(14.2±4.7) months. The ultrasoundcardiogram showed that all the ejection fraction(EF) of right ventricle improved compared with preoperation(P<0.05). The pulmonary artery systolic pressure(SPAP), both internal diameter and regurgitation volume of right ventricle were decreased(P<0.05). In the 310 patients, 302 patients(97.4%) were with the TR class 0-Ⅰ, 5 patients(1.6%) with class Ⅱ, 3 patients(1.0%) with class Ⅲ. There was no severe TR or no patient with reoperation. ConclusionThe MC3 ring is easy for planting and has good repeatability, which provides stable and satisfactory results for plasty of the tricuspid annulus with seldom residue or recurrent TR.
目的 研究磷脂酰肌醇3-激酶（PI3K）和磷酸化蛋白激酶B（p-Akt）在人膀胱尿路上皮癌组织中的表达特征及临床意义。 方法 2005年6月－2010年7月，采用免疫组织化学法检测40例膀胱尿路上皮癌组织及10例正常膀胱组织PI3K与p-Akt的表达，并对结果进行统计学分析。 结果 PI3K和p-Akt在正常膀胱黏膜组织阳性表达率均低于膀胱尿路上皮癌组织中，差异均有统计学意义（P＜0.05）。同一标本中PI3K和p-Akt的表达不具有相关性（r=0.051，P=0.747）。 结论 PI3K、p-Akt在膀胱尿路上皮癌中高表达，两者在膀胱尿路上皮癌中共同促其发展，但其在膀胱尿路上皮癌的预后和进展中的作用尚不明确。
ObjectiveTo explore the predictive value of systolic pulmonary artery pressure (SPAP) on autonomic nerve excitation in patients with valvular disease, so as to provide reference for the formulation of clinical intervention plans. Methods The clinical data of patients with valvular disease who received surgical treatment in the General Hospital of Northern Theater Command from August 28, 2020 to February 3, 2021 were prospectively collected. According to the standard deviation of normal-to-normal R-R intervals (SDNN) of the heart rate variability (HRV) of the long-range dynamic electrocardiogram (ECG) 7 days before the operation, the patients were divided into three groups: a sympathetic dominant (SE) group (SDNN≤50 ms), a balance group (50 ms<SDNN<100 ms) and a parasympathetic dominant (PSE) group (SDNN≥100 ms). The correlation between the changes of echocardiographic indexes and autonomic nerve excitation among the groups and the predictive values were analyzed. Results A total of 186 patients were enrolled, including 108 males and 78 females aged 55.92±11.99 years. There were 26 patients in the SE group, 104 patients in the balance group, and 56 patients in the PSE group. The left anteroposterior diameter (LAD), left ventricular end diastolic inner diameter, ratio of peak E to peak A of mitral valve (Em/Am), left ventricular end diastolic volume, left ventricular end systolic volume and SPAP in the SE group were higher than those in the balance group (P<0.05), while peak A of tricuspid valve (At) and left ventricular ejection fraction (LVEF) were lower than those in the balance group (P<0.05). The LAD and Em/Am in the balance group were significantly higher than those in the PSE group (P<0.05). Multivariate analysis showed that patients in the SE group had lower At (right atrial systolic function declines), lower LVEF and higher SPAP than those in the balance group (P=0.04, 0.04 and 0.00). When HRV increased and parasympathetic nerve was excited in patients with valvular disease, Em/Am decreased (left atrial function and/or left ventricular diastolic function declined) with a normal LAD. Pearson analysis showed that there was a linear negative correlation between SPAP and SDNN, with a coefficient of −0.348, indicating that the higher SPAP, the lower HRV and the more excited sympathetic nerve. Receiver operating characteristic curve showed that when SPAP≥45.50 mm Hg (1 mm Hg=0.133 kPa), the sensitivity and specificity of sympathetic excitation in patients with valvular disease were 84.60% and 63.70%, respectively. ConclusionParasympathetic excitation is an early manifestation of the disease, often accompanied by decreased left atrial function and/or left ventricular diastolic function. Sympathetic nerve excitation can be accompanied by the increase of SPAP and the decrease of left ventricular and right atrial systolic function. SPAP has a unique predictive value for the prediction of autonomic nerve excitation in patients with valvular disease.
Abstract:Objective To investigate clinical characteristics of patients with aortic valve disease and coronary artery disease （CAD）， and improve the pertinence of clinical management. Methods Clinical data of 78 patients who were older than 50 years and underwent isolated aortic valve replacement （AVR） in General Hospital of Shenyang Command from January to October 2010. All the patients were divided into two groups according to whether they had CAD：20 patients with CAD in the CAD group， including 19 males and 1 female with their average age of 64.6±8.3 years， and 58 patients without CAD in the control group， including 28 males and 30 females with their average age of 58.7±6.2 years. Clinical characteristics of AVR patients with CAD were analyzed by one-way analysis of variance and multiple factor analysis. Results One-way analysis showed that the incidence of old age （P=0.000）， male patients （P=0.004）， diabetes （P=0.004）， aortic regurgitation （P=0.034）， valvular lesion （P=0.028）， and postoperative mechanical ventilation time （P=0.008）of CAD group patients were significantly higher or longer than those of control group patients. Multiple factor analysis showed that independent clinical characteristics of AVR patients with CAD included male patients， old age， prolonged postoperative mechanical ventilation time， significantly decreased pulmonary arterial systolic pressure （PASP） at 6th postoperative month， and significant preoperative right ventricular diastolic dysfunction. Conclusion The screening age for possible CAD should be reasonably lowered if male patients with aortic valve disease have concomitant preoperative right ventricular diastolic dysfunction and/or tricuspid aortic valve degeneration. Patients undergoing isolated AVR need to take medications to reverse left ventricular remodeling for a long time to avoid severe arrhythmia.