Objective:To observe influence of simvastatin on differentiation ,proliferation ,migration and adhesion of marrow-derived smooth muscle progenitor cells (SPCs) and screen coated eluting stent drugs of new generation . Methods :The mononuclear cells (MNCs) were isolated from rat marrow by density gradient centrifugation method , and then plated on fibronectin-coated culture dishes ,after culture 8d ,marrow-derived SPCs were identified by α-smooth muscle actin (α-SMA) immunofluorescent staining and counted under inverted fluorescence microscope .The MNCs and adhesion cells were treated with simvastatin (0.01~10 μmol/L) respectively for 8 d and 24h .SPCs pro-liferation ,migration and adhesion were observed by Tritium thymidine (3 H-TdR) intake method ,modified Boyden chamber assay and adhesion assay .Results:Compared with control group (no simvastatin intervention ) ,0.01μmol/L simvastatin significantly inhibited the MNCs differentiation towards SPCs [ (85 ± 4) vs .(79 ± 5)] ,proliferation [ (4070 ± 184) vs .(3833 ± 126)] ,migration [ (44 ± 3) vs .(39 ± 3)] and adhesion of SPCs [ (59 ± 5) vs .(52 ± 4)] , P<0.05 all ,and number of SPCs significantly reduced along with simvastatin concentration increased (P<0.01) . Conclusion:Simvastatin could inhibit the differentiation ,proliferation ,migration and adhesion of marrow-derived smooth muscle progenitor cells .

Objective to investigate the relationship between the size of ostium secondary atrial septal defect (ASD) and the pulmonary arterial pressure (PAP) in children less than 5 years of age. Methods During the period from April 2000 to January 2011, a total of 189 child patients less than 5 years of age with ostium secondary ASD were admitted to General Hospital of Shenyang Military Command. Under general anaesthesia with ketamine cardiac catheterization was performed, PAP was measured, and percutaneous occlusion of ASD was carried out in all patients. The clinical indexes, including sex, age, body height, body weight, body surface area, diameter of ASD defect, heart- to- thorax ratio, the systolic, diastolic and mean pressure of the pulmonary artery, etc. were determined. The patients were followed up for one year and postoperative cardiac ultrasonography was performed to check the results. The patients were divided into groups according to the defect size. Results The 189 patients consisted of 77 males and 112 females with a male-to-female ratio of 1 ∶ 1.5. The mean age was (4.1 ± 0.9) years old, ranging from 2 to 5 years old. The mean weight was (17.2 ± 3.6) kg, ranging from 10.0 to 30.0 kg. The mean height was (104.9 ± 9.2) cm, ranging from 77 to 135 cm. The mean body surface area (BSA) was (0.71 ± 0.10) m2, ranging from 0.46 to 1.02 m2. The mean size of ASD was (12.6 ± 4.8) mm, ranging from 5 to 29 mm. The mean size of ASD, which was modified by BSA, was (18.0 ± 7.0) mm/m2, ranging from 5.3 to 38.9 mm/m2. The mean systolic PAP was (41.1 ± 8.9) mmHg with a range of 15 - 67 mmHg. The mean diastolic PAP was (16.8 ± 6.5) mmHg with a range of 3 - 45 mmHg. The mean PAP was (24.9 ± 6.7) mmHg with a range of 12 - 48 mmHg. One hundred and fifty- nine patients (89.4%) had pulmonary arterial hypertension (PAH) which was determined by right heart catheterization, but no patient showed PAH when the pulmonary arterial pressure was measured by echocardiography before the procedure as well as 1, 3, 6, 12 months after the procedure. No definite correlation existed between the size of ASD and the pulmonary artery pressure (P > 0.05). Conclusion Pulmonary artery pressure measured by right heart catheterization has no definite correlation with the size of ASD in children less than 5 years of age. Pulmonary artery pressure obtained from right heart catheterization is higher than that determined by cardiac ultrasonography, which may be caused by the effect of ketamine when general anaesthesia is used in performing right heart catheterization.

Objective To investigate the risk factors and early diagnosis of the severe thrombocytopenia complicating transcatheter ccclusion of patent ductus arteriosus ( PDA ) . Methods Between February, 2011 and May, 2015, 80 patients with patent ductus arteriosus underwent percutaneous intervention occlusion were studied. Results Average age were ( 17. 5 ± 17. 1 ) years, 63 were females (78. 8%), mean weight were (35. 6 ± 20. 2)kg (from 6. 0 to 75. 0 kg), mean body surface area (BSA) were (1. 09 ±0. 44) m2(from 0. 32 to 1. 91 m2). A bolus of heparin calcium (80 U/kg) was administered by intravenous injection. The mean diameters of patent ductus arteriosus were 4 mm (from 2 to 18 mm), and the mean diameters of occluders were 12 mm (from 6 to 30 mm). 14 patients were found to have severe thrombocytopenia (PLT count﹤100 × 109/L). The reduction rate of platelet in 12 of 14 patients was more than 19%. The diameters of all occluders were equal to or more than 14 mm, the mean diameters of patent ductus arteriosus were 10 mm ( from 6 to 18 mm) and the mean diameters of occluders were 18 mm ( from 14 to 30 mm). All the 14 patients started to present progressive decrease in PLT count since the second day post procedure. Taking the selected occluder diameter greater than 14 mm as cut-off points in diagnosis of severe thrombocytopenia, the sensitivity was 100%, specificity was 68%, the positive predictive value was 40%, and the negative predictive value was 100%. Combined with the postprocedural second day complete blood count analysis and the platelet count decreased by 10% as cut-off points in diagnosis of severe thrombocytopenia patients, the sensitivity was 93%, specificity was 67%, the positive predictive value was 65%, the negative predictive value was 93% . If taking the platelet count decreased by 7% on second day as cut-off points in diagnosis of severe thrombocytopenia patients, the sensitivity was 100%, specificity was 57%, the positive predictive value was 61%, the negative predictive value was 100% . Logistic regression analysis discovered that risk factors of severe thrombocytopenia after PDA are procedural platelet count and occluder diameter. Conclusions The risk factors of severe thrombocytopenia complicating transcatheter ccclusion of patent ductus arteriosus were the procedural reduction of platelet count and big occluder diameter. Patients with PDA who were inplanted with occluders equal to or bigger than 14 mm should retest the numbers of platelet on the second day after procedure and retest on third day if the numbers reduce on the second day, which may help in the prediction of severe thrombocytopenia.

Objective: To study the expressions of endothelin-1 in primary cultured atrial myocyte model at early stages of atrial fibrillation by rapid pacing. Methods: The primary rat atrial myocytes were cultured, in which a rapid paced cell model was established. The polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA) were applied to detect the messenger ribonucleic acid (mRNA) expression and secretion of endothelin-1 on 3 h, 6 h, 12 h and 24 h after rapid pacing. Results: Compared with before pacing, there were continuous and significant increase in expression of ET-1mRNA [ (0.31±0.02) vs. (0.52±0.07), (0.62±0.09), (0.75±0.05) ] and secretion of endothelin-1 [ (3.4±0.8) ng/L vs. (6.0±1.4) ng/L, (8.3±1.5) ng/L, (11.2±2.1) ng/L] on 6 h, 12 h and 24 h after rapid pacing, P<0.01 all. Conclusion 　 Expressions of ET-1mRNA and secretion of endothelin-1 significantly increase depend on time in early rapid pacing atrial myocytes, indicating that endothelin-1 participates in atrial pathological remodeling during atrial fibrillation.

This study was carried out to obtain the basic data for artificial culture of veiled lady mushroom (Dictyophora spp). The optimal conditions for the mycelial growth were 25degrees C and pH 5.0 for all isolates except the optimal temperature of 30degrees C for D. echinovolvata ASI 32002 and Phallus rugulosus . The optimal medium for Dictyophora spp. was PBA (potato bamboo sawdust extract agar) medium. The strain ASI 32002, D. echinovolvata , grew faster than. D. indusiata ASI 32003 and Phallus rugulosus ASI 25007 on the medium. Carbon sources such as glucose, maltose and inuline were favorable for stimulating a mycelial growth of the two strains of ASI 32002 and ASI 32003. Asparagine and glutamine appeared to be favorable to the strain ASI 32002 and ASI 32003, where as alanine, one of nitrogen source also favorable to the strain ASI 32002. The optimum C/N ratio of the two isolates of ASI 32002 and ASI 32003 was about 25 : 1 when 2% glucose as carbon source was mixed with the basal medium. While, in the case of 4% as carbon source, the optimum C/N ratio was about 30 : 1.
Agaricales* ; Alanine ; Asparagine ; Carbon ; Cultural Characteristics* ; Glucose ; Glutamine ; Hydrogen-Ion Concentration ; Inulin ; Maltose ; Nitrogen

Heat shock protein gp96 is a glycoprotein which was found several years ago. Besides its function as a molecular chaperone, it is also reported to play important roles both in innate immunity and adaptive immunity. Gp96 can stimulate the maturation of antigen presenting cells (especially dendritic cells) and the secretion of cytokines. Gp96 and its associated peptides could stimulate peptide specific cytotoxic T lymphocyte reaction (CTL), which was very promising in the designing of anti-virus and anti-tumor vaccines. However the expression level of whole length gp96 was relatively low in E. coli and the purity of gp96 are not very suitable for further study. We successfully cloned the carboxy terminal fragment (560aa-751aa) of murine gp96 into the pGEX-6p-1 vector and expressed in BL21 strain. This fragment contains the peptide binding domain and the dimerization domain. After purification, the recombinant fusion protein was cleaved with the PreScission Protease and analyzed by Gelfiltration. The results show that this fragment may be related to the dimerization of gp96 and make an foundation for further investigations of the protein.
Animals ; Antigens, Neoplasm ; genetics ; Blotting, Western ; Chromatography, Gel ; Cloning, Molecular ; Mice ; Peptide Fragments ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; isolation & purification

Myasthenia gravis (MG) is a disease of neuromuscular junction and mainly autoimmune in aetiology. The state of thymus is a critical determinant for the prognosis. In this retrospective review study, we aimed at clarifying the relationship between the mode of clinical presentation of MG and the radiopathological classification of the thymus. We identified patients with MG from the database of our medical center from 1988 – 2017. The patients were classified into two groups according to their clinical presentation: those with a typical presentation with diurnal variation, and those with an atypical presentation of persistent weakness or respiratory failure from the beginning. The underlying thymic state was categorized into six groups: normal, abnormal by imaging (if no operation was performed), hyperplasia, benign thymoma, cortical type thymoma, and malignant thymoma. In total, 227 patients (133 females and 94 males) were included in the analysis, of whom 68% were classified into the typical presentation group. The atypical presentation correlated significantly with thymic categories (p = 0.014) and sex (p = 0.026) but not age at onset (p = 0.232). The atypical presentation was more common in the male patients and in those with thymic carcinoma.

Objective To investigate the distribution of hepatitis delta virus (HDV) marker among hepatitis B virus (HBV) infected patients and to reveal its clinical significance.Method To collect the clinical data and sera samples of HBV infected patients and to detect HDAg,Anti-HDV as well as HBV infection markers by means of enzyme-linked immunosorbnent assay.These data combined with clinical diagnostic results and biochemical index were then analyzed.Result 462 samples of HBV infected patients were collected including 210 HBV carriers without symptom,175 chronic HBV infections,35 acute HBV infections and 42 liver fibrosis.The HDV infection rate was 4.8％ overall.The highest infection rate of 9.5％ was found in the group of liver fibrosis whereas the lower rate of 6.9％ was found in HBV chronic carriers.HDV infection rate was 7.8％ among the population of 40-60 years old,obviously higher than any other age groups.Conclusion HDV infection was significantly higher in the chronic HBV patients and liver fibrosis patients.Because HDV infection was highly associated with the progress of liver disease,we suggest the screen of HDV markers among hepatitis patients and discriminate whether the patient was co-infected with HDV.

This study investigated the regulatory role of nerve growth factor (NGF) in sirtuin 1 (SIRT1) expression in cholestatic livers. We evaluated the expression of NGF and its cognate receptors in human livers with hepatolithiasis and the effects of NGF therapy on liver injury and hepatic SIRT1 expression in a bile duct ligation (BDL) mouse model. Histopathological and molecular analyses showed that the hepatocytes of human diseased livers expressed NGF, proNGF (a precursor of NGF), TrkA and p75NTR, whereas only p75NTR was upregulated in hepatolithiasis, compared with non-hepatolithiasis livers. In the BDL model without NGF therapy, p75NTR, but not TrkA antagonism, significantly deteriorated BDL-induced liver injury. By contrast, the hepatoprotective effect of NGF was abrogated only by TrkA and not by p75NTR antagonism in animals receiving NGF therapy. Intriguingly, a positive correlation between hepatic SIRT1 and NGF expression was found in human livers. In vitro studies demonstrated that NGF upregulated SIRT1 expression in mouse livers and human Huh-7 and rodent hepatocytes. Both NGF and proNGF induced protective effects against hydrogen peroxide-induced cytotoxicity in Huh-7 cells, whereas inhibition of TrkA and p75NTR activity prevented oxidative cell death. Mechanistically, NGF, but not proNGF, upregulated SIRT1 expression in human Huh-7 and rodent hepatocytes via nuclear factor (NF)-κB activity, whereas NGF-induced phosphoinositide-3 kinase/Akt, extracellular signal–regulated kinase and NF-κB signaling and SIRT1 activity were involved in its hepatoprotective effects against oxidative injury. These findings suggest that pharmacological manipulation of the NGF/SIRT1 axis might serve as a novel approach for the treatment of cholestatic disease.
Animals ; Bile Ducts ; Cell Death ; Cholestasis ; Hepatocytes ; Humans ; Hydrogen ; In Vitro Techniques ; Ligation ; Liver ; Mice ; Nerve Growth Factor ; Phosphotransferases ; Rodentia ; Sirtuin 1

Atrioventricular Block ; etiology ; Cardiac Catheterization ; adverse effects ; Heart Septal Defects, Ventricular ; therapy ; Hematologic Diseases ; etiology ; Hemolysis ; Humans