BACKGROUND: Both density gradient centrifugation and adherence method arc frequently used to isolate bone marrow mesenchymal stem cells (BMSCs).OBJECTIVE: To investigate the approaches to isolate, culture and identify the rabbit BMSCs in vitro by the combination of den,ity gradient centrifugation and adherence method. DESIGN, TIME AND SETTING: Contrast cytological study, which was performed in Central Laboratory of Shanghai 6th People's Hospital between October 2007 and March 2008.MATERIALS: Six 2-week-old rabbits were selected for BMSCs preparation and primary culture; Percoll separating medium (1.073 kg/L) was also used for this study.METHODS: BMSCs were separated and purified with Percoll separating medium by density gradient centrifugation and adherence method. The three-, five-, seven-, and nine-passage BMSCs were counted for growth curve. MAIN OUTCOME MEASURES: Morphological features and growth states of primary and passage cells were observed under inverted microscope. Indirect immunofluorescence of CD44 and CD34 antibodies were used to examine the stem cells. CD44 staining was positive, and CD34 staining was negative, suggesting the extracting and purifying cells were BMSCs. RESULTS: The passage BMSCs were uniformly distributed like fusiform shape, which were more uniform than primary cultured cells. The BMSCs grew productively and proliferated rapidly; meanwhile, the nucleolus was clear, caryopla.sm was in a large proportion, morphological features were uniform, ceils like bostrychoid or whirlpool were arranged parallelly, and the five-pa.ssage cells were not changed remarkably. Proliferation was decreased gradually with the passage increasing; especially, the proliferation of three-five-passage cells was the strongest. The separated cells expressed CD44 but not CD34. CONCLUSION: High-purified rabbit BMSCs are obtained by both density gradient centrifugation and adherence method.

Objective · To explore the correlation between variants located in 3' untranslated regions (3'UTR) of NOTCH1 and JAG1 genes and conotruncal heart defects (CTD). Methods · Six hundred CTD children without 22q11 deletion and three hundred healthy children were enrolled in this hospital-based case-control study. Variants located in the 3'UTR regions of NOTCH1 and JAG1 genes were detected by high-throughput sequencing. The accuracy of the variants were verified by PCR and Sanger sequencing. Online software PicTar, TargetScan and microRNA.org were used to make functional predictions. Results · One mutation and three SNPs were found in the 3'UTR of NOTCH1. Three mutations and six SNPs were found in the 3'UTR of JAG1. The genotypic distributions of two SNPs (rs3840074 and rs8708) located in JAG13'UTR between CTD group and the controls were statistically significant (both P<0.05). Results of prediction showed that all the four mutations and two meaningful SNPs could bind to microRNA. Conclusion · The variants located in 3'UTR regions of NOTCH1 and JAG1 genes may be related to the occurrence of CTD.

Objective To compare the short and long term effect of knee osteoarthritis treatment with two therapeutic methods which are simple high cut of fibula and arthroscopic lavage debridement combined with fibular osteotomy.Methods Data of 45 consecutive patients with knee osteoarthritis were retrospectively observed and selected,and then they were divided into two groups according to the operation method.23 patients with simple high cut of fibula were selected as the control group,and 22 patients with arthroscopic lavage debridement combined with fibular osteotomy were selected as the observation group.Results The VAS and AKS scores of the control group at the time of 3m,6m,1 2m were (4.1 3 ±0.76)points,(1 07.04 ±21 .53)points;(2.70 ±0.64)points,(1 41 .1 7 ±1 2.57)points;(2.43 ±0.79)points,(1 48.26 ±5.81 )points;and the scores of the observation group were (3.45 ± 0.60)points,(1 26.64 ±1 4.24)points,(2.70 ±0.73)points,(1 46.45 ±7.26)points,(2.41 ±0.85)points and (1 48.26 ±5.81 )points.The differences between the two groups were statistically significant at the time of 3 months after surgery(t =3.32,-3.62,all P <0.05),the effect of observation group was better than the control group;but the other indices had no statistically significant differences (t =0.51 ,-1 .72,0.1 1 ,-0.20,all P >0.05 ). Conclusion The simple high cut of fibula with small trauma,less cost,less surgical difficulty,and long -term clini-cal curative effect is not less than the arthroscopic lavage and debridement combined with fibular osteotomy.There-fore,it should be carried out in the primary hospitals widely.

Objective To investigate the relationship between high sensitive C reactive protein(hsCRP) and monocyte chemotactic protein-1 (MCP-1) in patients with metabolic syndrome(MS).Methods Sixty-two patients with MS and 52 normal healthy individuals (NS) were recruited in the study.And the levels of hs-CRP and MCP-1 were detected by ELISA.The severity of Insulin resistance was calculated by homeostasismodel assessment of insulin resistance(HOMA-IR).The monocytes in blood were isolated,cultured and stimulated by different concentrations of CRP.MCP-lexpression of monocytes was measured by real-time PCR.Results Patients with MS displayed significantly higher serum levels of hs-CRP and MCP-1 compared with healthy control group [(80.26 ±35.52) nmol/L vs (3.12 ±2.55) nmol/L for hs-CRP,P ＜0.01 ; (15.35 ± 10.12) nmol/L vs (9.76 ± 6.15) nmol/L for MCP-1,P ＜ 0.05].There was significantly positive correlation between hs-CRP and MCP-1.Serum hs-CRP,MCP-1and insulin resistance index were positively correlated,respectively.In vitro experiments indicated that CRP could up-regulate MCP-1 expression of monocytes in a concentration-dependent way.When the concentration of CRP was 176nmol/L,the expression of MCP-1 could reach 105％ compared with non-stimulated group.Conclusion With the severity of the inflammatory response and enhanced insulin resistance,elevated serum CRP level may result in MCP-1 expression in patients with metabolic syndrome.

Objective To observe the expression of PECAM-1 in colorectal cancer in rat and to explore the mechanism of lymphatic metastasis of tumor. Methods Wistar rats were used to make the animal model of primary colorectal cancer,by using HE staining,the cancer type and progression of cancer were devided.By immunohistochemistry staining,the expression of PECAM-1 in tumor tissue,blood vessels and lymphatic vessels were observed. Results PECAM-1 expression in blood vessels and lymphatic vessels were observed in normal tissue,while its expression on tumor tissue and lymphatic vessels decreased according to the cancer progression.Conclusion PECAM-1 may be involved in the early process of tumor cells adhering to the endothelium;decreased expresson of PECAM-1 on lymphatics may be related with lymphangiogenesis and the opening of endothelial junction of lymphatic vessels.

Objective To study the ultrastructural features of initial lymphatics of gastric carcinoma,to discuss the metastasis mechanism of malignant cells by lpmphatics. Methods The postoperative samples of patients with gastric cancer were obtained in center,peripheral and normal region of cancer.Common Resin-embedded specimens were made.Lymphatics were located by hemithin sections under light microscope and observed by ultrathin sections under electron microscope. Results There were no lymphatics in carcinomatous centre region.Compared with normal region,there were more initial lymphatics in peripheral region of gastric carcinoma.The lumina were dilated and irregular and the endothelium were destroyed;The organelle of endothelial cells changed obviously.The distribution and morphology of the lymphatics did not changed evidently in the normal region.Conclusion These results suggest that amount,ultrastructure and morphology of lymphatics changed in peripheral region of carcinoma;Gastric cancer cells may invade lymphatics via destroyed parts of endothelium or interendothelial openings.

Objective To explore the role of splenectomy in the prevention and treatment of small-for-size liver in rat models, as well as its pathophysiologic mechanism in the development of a small-for-size syndrome (SFSS). Methods The models of sham-operation and 80 % partial hepatectomy (PH) were used in rats. In the experiment group splenectomy was performed following 80% PH. The concentrations of serum tumor necrosis factor (TNF-α), the content of NF-cB p65 in liver nuclear extracts, the expression of TNF-α, intercellular adhesion molecular (ICAM-1), and proliferating cell nuclear antigen (PCNA) transcripts, the activities of serum aspartate transaminase (AST), alanine transaminase (ALT), lactate dehydrogenase (LDH), total bilirubin (TB), albumin (Alb) cholinesterase (CHE), and liver myeloperoxidase (MPO) were analyzed. Portal venous pressures (PVP),incidence of SFSS,and one-wk survival rate were measured. Results In the control rats,The PVP was obviously elevated immediately after PH. The level of NF-κB p65 was obviously increased at the first h and peaked at about 3rd h postoperatively. The transcription of TNF-α and ICAM-1 and the release of serum TNF-α were significantly increased 3 h after PH. Capillary endothelial cells of the livers strongly expressed ICAM-1 24 h after PH. Splenectomy significantly reduced the PVP and the content of NF-κB p65 in the livers in concurrence with the expression of TNF-α and ICAM-1 gene as well as the activity of MPO at the corresponding time points after PH (P＜0. 05), while increased the expression of PCNA gene (P＜0. 05). Administration of splenectomy resulted in a statistically significant decrease in AST, ALT, LDH, TB, the incidence of SFSS and increase in one-wk survival rate (P ＜ 0.05 ). Conclusion Splenectomy alleviates liver injury and promotes liver regeneration in small-for-size liver rats by reducing portal vein perfusion and pressure,and suppressing NFκB activation and subsequent expression of proinflammatory mediators.

This paper presents a way to carry out the RAC technology in hospital, researches the key technologies of RAC, the result shows that performance of database is greatly improved by RAC technology.
Databases, Factual ; Hospital Information Systems ; Integrated Advanced Information Management Systems

Objective To explore the pathophysiologic mechanism of the development of a small-for-size syndrome(SPSS) and the role of splenectomy in the prevention and treatment of SFSS.Methods The rat models of sham-operation and 80% partial hepatectomy were established.Totally 144 rats were randomly divided into 3 groups:1)splenectomy group:splenectomy was performed following 80% partial hepatectomy;2)control group:80% partial hepatectomy was performed;3)sham group:no hepatectomy was performed.After the operation,we examined the portal venous pressures(PVP),tumor necrosis factor(TNF-α) and proliferating cell nuclear antigen(PCNA) expression,the activity of myeloperoxidase(MPO),liver function and explored the prevalence of SFSS.Results Compared with the sham group,the PVP of the rats in the control group obviously elevated after hepatectomy,and the expression level of TNF-a and the activity of MPO in the liver significantly increased(P<0.05).Compared with the control group,the PVP,the expression of TNF-a in the livers and the activity of MPO at the corresponding time points after hepatectomy in the splenectomy group significantly decreased,while the expression of PCNA in-creased(P<0.05).Administration of splenectomy resulted in a statistically significant decrease in aspartate transaminase(AST),alanine transaminase(ALT),lactate dehydrogenase(LDH),total bilirubin.and the incidence of SFSS(P<0.05).Conclusion Splenectomy could alleviate liver injury,promote liver regeneration in small-for-size liver rats by reducing portal vein perfusion and pressure and the subsequent expression of proinflammatory mediators.

Objective To explore the predictive value of serum N-terminal pro-brain natriuretic peptide (NT-proBNP) level in anthracycline-based chemotherapy-related cardiotoxicity of breast cancers.Methods A total of 135 breast cancer patients was analyzed if NT-proBNP was associated with chemotherapy-related cardiotoxicity.The level of NT-proBNP in the diagnosis of cardiotoxicity was assessed.Results A total of 22 patients (16.29％) had subsequent claims for cardiotoxicity events.NT-proBNP in cardiotoxicity group was significantly higher than that non-cardiotoxicity group (P ＜ 0.05).According to receiver operating characteristic (ROC) curve, the cut-off value of NT-proBNP was set at 350 pg/ml, specificity and sensitivity were 70.32％ and 82.58％ , respectively.Positive and negative predictive values were 78.12％ and 65.45％, respectively.Conclusions The present study is to confirm excellent clinical value of NTproBNP on cardiotoxicity.The level of NT-proBNP for early detection of cardiotoxicity has good prospects for high risk patients.