Amniotic fluid-derived mesenchymal stem cells(AF-MSCs)are multipotential,nonhemato-poietic progenitor cells that have an inherent ability to self -renew and to differentiate into many cell types ,inclu-ding osteoblasts,chondrocytes and adipocytes.Despite their high proliferation rate,no in vivo tumorigenicity has been reported and their karyotype was normal even after extensive expansion in culture .Several studies have dem-onstrated the inherent tumortropic properties of MSCs and their successful use as cellular vehicles of therapeutic agents for the treatment of several types of cancer .This article reviews recent articles on studying the role of amni-otic fluid-derived mesenchymal stem cells in cancer therapy .

Objective To study clinical value of blood glucose, AFP and cholinesterase levels on the diagnosis and prognosis of severe hepatitis. Methods Retrospective analysis of chronic hepatitis B, cirrhosis, chronic severe hepatitis, subacute severe hepatitis was carried out. In severe hepatitis patients, between the improved group and death group, blood glucose, AFP, cholinesterase differences, evaluate blood glucose, alpha-fetoprotein, cholinesterase levels on the diagnosis and prognosis of severe hepatitis value were compared. Results Subacute severe hepatitis, chronic severe hepatitis patients and blood cholinesterase levels were significantly lower than that in patients with chronic hepatitis B and liver cirrhosis(all P ＜ 0.05) ;and alpha-fetoprotein level was significantly higher than the level of chronic hepatitis B patients(P ＜ 0. 01) ;improved levels of blood glucose in patients with severe hepatitis, AFP, cholinesterase levels were significantly higher than the level of death group (all P ＜ 0.05). Conclusion Blood glucose, AFP, cholinesterase levels of liver disease progression and prognosis were important to determine the value.

AIM:Neural stem cells can be induced to differentiate into various types of neural cells such as neurons and neuroglia cells,but the technique of depuration and cultivation does not consummate.This article determines the optimal culture technique of neural stem cells by different culture concentrations and passage methods. METHODS:Experiments were conducted from May to December 2006 at Laboratory of Transplantation Immunity of Sichuan University.①Clean pregnant female rats(embryonic age range from 12-16 days)and the disposition of animal met ethical standard.②The cerebral cortex of rat embryos were collected,and digested with trypsin and ethylenediamine tetraacetic acid mixture to obtain signal cell suspension.They were cultured in serum-free medium (DMEM/F12 medium containing B27,basic fibroblast growth factor and epidermal growth factor).The 3~(rd)passage cells were collected,and incubated at 1?10~7 L~(-1),1?10~8 L~(-1),1?10~9 L~(-1),1?10~(10)L~(-1),respectively.In addition,neural stem cells were collected 7-10 days after primary culture to harvest formative cell masses.Mechanical blow refers to soft blowing with haustorial tube from thick to thin after centrifugation,or sterile syringe with No.5 pinhead blowing cells when the blow was 5 times.Bubble production was avoided during the operation.Trypsin aspiration combined with mechanical blow refers to trypsin was added after centrifugation,at 37℃for 10 minutes,the neural stem cells were lightly blown with haustorial tube polished with flame or blown with sterile syringe with No.5 pinhead,and then fetal bovine serum was added to stop digestion.③The growth characteristic of the 3~(rd)passage cells at different culture concentration was observed and proliferation was measured at days 1,3,5 and 7 by MTT assay.The neural clone spheres of subcultured was counted to determine the optimal passage way.Immunofluorescence was carried out to detect nestin(special marker to neural stem cells),BrdU,neurone specific enolase,glial fibrillary acidic protein. RESULTS:①Growth characteristics and identification of rat embryonic neural stem cells in vitro:The dissociated neural stem cells from the cerebral cortex of rat embryos were continuously harvested and purified by suspension cultures to get the daughter cell clone.Nestin positive cells could be found in the neurospheres and after attachment they could differentiate into neurone specific enolase and glial fibrillary acidic protein positive cells,and immunofluorescence showed a great of BrdU-positive cells.②The effect of different incubated number of neural stem cells on proliferation:When the neural stem cells planted at the concentration of 1?10~9 L~(-1),the growth rate of the cells was the highest of all concentrations.The number of clone spheres exceeded others at concentration 1?10~7 L~(-1),1?10~8 L~(-1)and 1?10~10 L~(-1)(P

Objective To study the application of multidector spiral CT(MSCT) in the diagnosis of pulmonary embolism(PE).Methods The clinical and MSCTPA data of 15 cases with suspected PE were analyzed retrospectively.Results Of 15 cases,14 cases(93.3%)were diagnosed as PE.849 branches(82%) of pulmonary arteries were showed in 1035 branches.PE was detected in 130 branches(15.3%) of 849 branches of pulmonary arteries.438 branches(73%) were showed in 600 branches of subsegmental pulmonary arteries,PE was showed in 43 branches(9.8%)of 438 branches.Direct signs of PE included central filling defect in 3 cases;eccentric filling defect(n=9);embolism attached to the wall of host artery(n=2) and total occlusion of the pulmonary arteries(n=3).Indirect signs includedmosaic sign in 1 case;subpleural infarction(n=3);pulmonary hypertension(n=5);Westermark sign(n=8) and pleural effusion(n=6).Conclusion MSCTPA is a fast,effective,security and non-invasive diagnostic method for PE,it may replace the pulmonaryangiography and will be the first choice for diagnosis of PE.

Objective To study the application value of chest low-dose spiral CT for the screening of lung cancer in physical examinees . Methods The data of physical examinees that underwent chest low-dose CT for screening of lung cancer from January 2013 to January 2016 was analyzed, the index included the correlation between age and sex with the detection rate of lung nodules and the detection rate of lung cancer, pathological types and clinical stages of lung cancer and so on. Results 9762 physical examinees underwent chest low-dose spiral CT, the detection rates of pulmonary nodules and lung cancer were 58.16%(5678/9762) and 0.34 % (33/9762) respectively. The detection rate of lung cancer increased with the age, the highest detection rate [1.13 % (8/709)] of lung cancer was found in 80 years old and above, the rate in 60-69 years old group was 0.36%(6/1641). In 33 cases of lung cancer, there were 22 cases in stage Ⅰ, 5 cases in stageⅢ, and 6 cases in stage Ⅳ. Early diagnosis rate was 66.67 % (22/33). Compared with the conventional thoracic spiral CT, the low-dose chest CT had loner radiation dose (P<0.05), and the image can meet the diagnostic requirement. Conclusions Chest low-dose spiral CT has great advantages in the detection of pulmonary nodules and early stage lung cancer. The application of low-dose CT in physical examinees, especially in high-risk groups of lung cancer, can improve the positive rate of lung cancer screening.

Objective To investigate the clinical efficacy of percutaneous vertebroplasty (PVP)for vertebral metastases.Methods 40 cases with 46 vertebral body metastases underwent PVP under the guidance of fluoroscopy or CT.The analgesic effect,vertebral body stability and complications after operation were observed.Results The successful rate of puncture was 100%.The pain relief was demonstrated in all cases(complete relief in 30 cases,partial relief in 8 cases,mld relief in 2 cases) after operation 1 to 3 days.All involved vertebral bodies had good stability.Imaging analysis demonstrated that the metastatic area was completely well-distributed filling and reinforce by polymethymethacrylate(PMMA) in 26 vertebra of 24 cases.The other 20 vertebral of 16 cases were great part filled by PMMA.There were no severe complications both during operation and post-operation.The treating effect was satisfying in the following 1 to 3 months.Conclusion PVP is one of the efficacious way to alleviate pain of vertebral metastases,and it can reinforce the stAbility of involved vertebral body.

AIM:To observe the change of nitric oxide(NO)and hydrogen sulfide(H_2S)in blood and lung homogenate of hypoxic pulmonary hypertension(HPH)rat model,and to discuss the meaning of inhalation sodium nitrite and these factors in the treatment of HPH. METHODS:Fifty healthy male Wistar rats were assigned randomly into 5 groups(10 rats each):normoxia control group(NC),normoxia sodium nitrite group(NNI),hypoxic control group(HC),hypoxic normal saline group(HNS)and hypoxic sodium nitrite group(HNI). The mean pulmonary arterial pressure(mPAP),weight of right ventricle,weight of left ventricle plus septum,and the ratio of the weight of right ventricle to that of left ventricle plus septum(right ventricle hypertrophy index,RVHI)were also determined. The serum level of NO and plasma level of H_2S were measured,and at the same time the levels of NO in the lung homogenate were detected. The structures in pulmonary arteries were examined using optical microscope. RESULTS:After model established,compared to that in the normoxia groups,the body weight decreased significantly in hypoxia groups(P<0.05),although no difference of body weight in five groups before producing model was observed. Compared to that in normoxia groups,the levels of mPAP and RVHI increased significantly in hypoxia groups,and compared to that in hypoxia control groups and hypoxia normal saline group,mPAP and RVHI levels decreased significantly in hypoxia sodium nitrite group(P<0.05). Compared to that in normoxia groups,the serum level of NO decreased significantly in hypoxia groups(P<0.05). NO level in lung homogenate decreased significantly in hypoxia control group and hypoxia normal saline group as compared to that in normoxia groups(P<0.05),and no obvious difference between hypoxic sodium nitrite group and normoxia groups was found. The plasma level of H_2S was decreased significantly in hypoxia groups(P<0.05)as compared to that in normoxia groups. H_2S level increased significantly in hypoxia sodium nitrite group as compared to that in hypoxia control groups and hypoxia normal saline group(P<0.05). Observation under optical microscope,the lumen structure of lung in normoxia control group was normal. No significant change in normoxia sodium nitrite group was found. The proliferation of smooth muscle cells(SMCs),the collagen fiber deposition in the vessel wall and every caliber thickening was observed in hypoxic control group. The same changes were also observed in hypoxic normal saline group. The thickened caliber was relieved significantly in hypoxic nitrite group. CONCLUSION:Pulmonary hypertension and right ventricle reconstitution can be relieved by inhalation of sodium nitrite,and can be regulated by the level of NO and H_2S in rats. Above all,inhalation of sodium nitrite may degrade HPH directly or by affecting the externalization and synthesizing of gas signaling molecule indirectly.

BACKGROUND: Hyperpolarization-activated cyclic nucleotide-gated (HCN) current plays an important role in regulating heart spontaneous pulsation.OBJECTIVE: To observe target gene expression and electrophysiological characteristics of pig bone marrow mesenchymal stem cells (BMSCs) transfected with hyperpolarization-activated cyclic nucleotide-gated channel 2 (HCN2) gene recombinant adenovirus.DESIGN, TIME AND SETTING: Cell-gene in vitro study was performed at the Laboratory of Thoracic and Cardiovascular Surgery,Second Military Medical University of Chinese PLA from July 2007 to March 2008.MATERIALS: Yorkshire pig was supplied by Animal Institute, Second Military Medical University of Chinese PLA. HCN2 plasmid was presented by Professor Dario DiFrancesco from Italy. Recombinant adenovirus Ad.HCN2 was constructed and stored using Ad5 in this laboratory.METHODS: Pig BMSCs were isolated with combination of gradient centrifugation of Percoll and adherent treatment in vitro.Ad.HCN2 was transfected at multiplicity of infection=50. We also set non-transfection and transfected Ad.Null groups.MAIN OUTCOME MEASURES: Expression of HCN2 mRNA was detected with RT-PCR, and expression of HCN2 channel protein was examined with immunofluorescent staining. Electrophysiology of HCN2 channel protein was measured with whole-cell patch clamp.RESULTS: No amplified fragments were found in the non-transfection and transfected Ad.Null groups, but amplified fragments were determined at 250-500 bp following Ad.HCN2 amplification, which was the same as plasmid carrying HCN2 gene. Staining strength of cell nuclei following transfection was significantly weaken compared with cell membrane and plasma, which showed identical distribution as HCN2 protein. No HCN2 protein was detected in the non-transfection and transfected Ad.Null groups.Pacemaker current could be recorded with a whole-cell patch clamp. It was fully activated around -140 mV with an activation threshold of -60 mV, presenting voltage dependence. CsCI (4 mmol/L) reversibly blocked the inward currents. No pacemaker current was detected in the non-transfection and transfected Ad.Null groups.CONCLUSION: The HCN2 recombinant adenovirus carrier was transferred into serial subcultivation porcine bone marrow mesenchymal stem cells. HCN2 channel protein has been expressing. Pacemaker current could be recorded with a whole-cell patch clamp.

The study is designed to investigate the progress in cardiac auscultation proficiency of residents after training program.Thirty eight residents were selected to test their ability of cardiac auscultation.There were 13 cardiac events(or murmurs)in 15 patients,and the auscultation proficiency was expressed as the percentage of recognizing cardiac events accurately.The results showed that the highest auscultation proficiency was whole systolic and diastolic murmurs(up to 79 percent and 61 percent respectively)before training.The percentage of recognizing continuous murmurs,click,mitral stenosis and regurgitation,aortic stenosis increased significantly after training program(P＜0.05).

Aim To evaluate the pharmacological effects of sodi um ferulate on asthmatic guinea pigs. Methods Guinea pigs were sensit ized by ovalbumin. Two weeks later,ment of Sodium ferulate were injected every d ay five minutes before inhale, the guinea pigs were killed 7 days later. Granul ocyte-macrophage colony-stimulating factor (GM-CSF), interleukin-4 (IL-4), interleukin-5(IL-5), Eosinophil cationic protein (ECP) in serum were measured by Enzyme-linked immunosorbent assay(ELISA) and fluoroimmunoassay method, the e osinophils (EOSs) were counted and the phosphodiesterase (PDE) in the bronchoalv eolar lavage fluid(BALF) were determined by inverse high-performance liquid chr omatography (HPLC) method. Result After treatment with sodium f erulate for one week at the dose of 25,50,100 mg?kg -1 respectively, th e GM-CSF, IL-4, IL-5 contents were inhibited as compared with the untreated g roup in a dose dependent manner.The count of eosinophil was decreased.The activi ty of PDE of cell in BALF and the level of ECP were decreased.Conclusion Sodium ferulate attenuates the GM-CSF, IL-4, IL-5 in serum and eosi nophils and the activity of PDE and ECP and has protective effect in asthmatic g uinea pig.