BACKGROUND:The benefit of cell therapy may be partly due to the secretion of angiogenic and antiapoptotic growth factors.Whether amniotic fluid stem cells (AFS) could secrete some growth factors requires further studies.OBJECTIVE:To isolate and culture AFS cells,and explore the angiogenic or antiapoptotic effect of cytokines secreted by AFS on endothelial cells.DESIGN,TIME AND SETTING:A in vitro cytological experiment was performed at the Institute of Hypertensive Disease,First Affiliated Hospital,Nanchang University from December 2008 to June 2009.MATERIALS:Term amniotic fluid of ten samples,50 mL/case,was obtained following caesarean delivery.The umbilical vein was used to isolate endothelial cells.Written informed content was obtained from all women.METHODS:AFS isolated from human amniotic fluid was cultured and digested by trypsin at confluence of 80%.The third passage of cells at a density of 5×10~8/L were divided into two groups:hypoxia group:the cells were cultured in 2% O_2 + 5% CO_2 +93% N_2;normal group:the cells were cultured in 5% CO_2 + 95% air.Two groups were cultured at 37 ℃ for 24 hours.The supematant of two groups was collected.The second passage of human umbilical vein endothelial cells cultured in vitro was collected and seeded onto 12-well culture plate at a density of 2×10~4 cells/well,and divided into 3 groups:control group was cultured in 2 mL EBM-2 containing 5% fetal bovine serum (FBS);normal group was cultured in 1 mL EBM-2 containing 5% FBS and 1 mL AFS cell culture solution;hypoxia group was cultured in 1 mL EBM-2 containing 5% FBS and 1 mL hypoxia AFS cell culture solution for 3 days,followed by incubation with 10 μg/L tumor necrosis factor (TNF)-α.MAIN OUTCOME MEASURES:AFS surface phenotype was examined by flow cytometry;the secretion level and mRNA expression of vascular endothelial cell growth factor (VEGF) and hepatocyte growth factor (HGF) were examined by ELISA or RT-PCR.The proliferation and apoptotic rates of endothelial cells were examined.RESULTS:AFS cells were long fusiform-shaped and arranged radially after 7 days of culture.The third passage of AFS cells expressed CD29 and CD105 while did not express CD34.AFS cells of normal culture secreted VEGF and HGF;AFS cells of hypoxia condition significantly increased secrete of VEGF (P＜0.01),and VEGF mRNA expression was significantly upregulated (P＜0.05),while HGF and mRNA expression remained unchanged (P＞0.05).Compared with control group,the number of endothelial cells was significantly increased in normal and hypoxia AFS cell groups after 3 days of culture (P＜0.05).After cocultured with TNF-α for 24 hours,the apoptosis rates of endothelial cells in AFS-conditioned medium was significantly decreased (P ＜ 0.05),and the change degree of hypoxia AFS cell group was greater than normal AFS cell group (P ＜ 0.05).CONCLUSION:AFS can secrete cytokines such as VEGF and HGF.Moreover,it significantly promotes endothelial cells proliferation and inhibits apoptosis.Under hypoxia condition,the secretion of VEGF from AFS cells is increased,and the effects on endothelial cells proliferation and apeptosis are enhanced.

Objective To investigate the effect of the Senlm of patients with congestive heart failure(CHF) on the apoptosis of endothelial progentior eelts(EPCs)and protective role of carvedil01.Method Totally 20 patientswithCHF、^mre enrolledfromDepartment ofCardiology ofthe FirstHospital Affdiated ofNanchangUniversity between June and August 2007.Serum WflflL isolatedfrom blood ofthe patients.1lononuelear cells were isolated from blood of20 healthy volunteers by density gradient centrifugation.Attaching cells wel-e harvested after culture for 7 relays and the EPCs were anyzea by flow cytometry and immunofluorescenee method.EPCs were divided into 4 groups:group A,cultured in sE-of volunteers;group B,cultured in serum of CHF;group C,cultured in leNlm of CFIF and Carvedilol;group D,cultured in,erum of CI-IF and metoproM,After euhure for 24 hours,annex.inV/PI wss used to investigate the apoptosis rate ofEPCs and the activity ofcllspafle 3 were exanlilled.ELISA Wills used to detect the level of flerum TNF-a of patients with CHF.Multiple group compalrion Wills conducted with variance analvsis and q test.Results The apoptosis rate and easpase 3 activity of EPCs in group B[(7.5 4-0.9)%v8.(3.0 4-0.25)]were significantly ineased compared with group A[(3.4±0.7)%VS.(1.37±0.2),P

AIM: To investigate the effect and the mechanism of apolipoprotein (a) [apo (a) ] on proliferation of vascular smooth muscle cells ( VSMCs). METHODS: All VSMCs used in experiments were serial subcultured from primary cells and were identified by immunohistochemistry staining of a - actin. Cell growth assay was observed as cell counting and MTT assay. Western blotting was also employed to detect the related mechanism. RESULTS: All cells used in experiments were confirmed as VSMCs. Although apo (a) enhanced VSMCs proliferation, this effect was attenuated by anti -integrin α_vβ_3, LM609.Use these reagents alone had no effect on VSMCs growth. The results of Western blotting demonstrated that focal adhesion kinase (FAK) was activated by apo (a) and the expression of total or phosphorylated transforming growth factor β_1 (TGF -β_1) was also decreased. However, these effects described above were all blocked by LM609.CONCLUSION: Apolipoprotein (a) enhances VSMCs proliferation and this effect is mediated by integrin α_vβ_3, which activates FAK and attenuates TGF - β_1 and phospho -TGF - β_1 expression.

Objective PSGL-1 is specifically expressed in leucocytes.The aim of this study was to explore the changes of myeloid-derived suppressor cells (MDSCs) in the spleen and bone marrow in PSGL-1-deficient mice.Methods PSGL-1 -/-mice were used in the experiment.After identification of the offsprings, flow cytometry was used to test the expression of CD11b and Gr-1 in C57 and PSGL-1 -/-mice.Results Compared with the C57 mice, the expression of MDSCs was up-regulated in the PSGL-1-deficient mice ( P <0.001).Conclusion The expression of MDSCs is upregulated in PSGl-1-deficient mice.

Objective Shkbp is also called Shkbp1,can competitively inhibit binding CIN85 and c-Cbl,thereby blocking the epidermal growth factor receptor (EGFR) endocytosis and degradation,to play a role in tumor promotion.This study aims to explore the changes in blood cell classification and T cell subsets in blood,bone marrow,and spleen in Shkbp1-deletion (Shkbp-1-/-) mice.Methods Shkbp-1-/-transgenic mice were identified by PCR genotyping.Blood cell classification was performed using an automatic classification system.Flow cytometry was used to detect the T lymphocyte subsets in the blood,bone marrow,and spleen of Shkbp-1-/-and control mice.Results Routine blood examination showed that neutrophils and eosinophils tended to increase and showing significant differences,and there was no significant difference in lymphocytes.The flow cytometry results showed that there was a decrease of CD4+CD8+ double positive cells and increase of bone marrow CD3+ and CD4+ cells in the control group.However,there was a decreasing trend of CD3+,CD4+,CD8+,and CD4+CD8+ cells in the spleen tissues.Conclusions Shkbp1 is involved in the maturation and differentiation of blood cells,and affects the number of immune cells.This study lays a foundation for the study of how Shkbp1 is involved in the differentiation of blood cells.

Objective To investigate the association between plasma lipoprotein (a) [LP (a)] concentra-tion and in-stent restenosis after coronary stent implantation. Methods 152 patients with successful elective coro-nary stont implantation and percutancous transluminal coronary angioplasty (PICA) undergoing foUow-up angiogra-phy were retrospectively analyzed. These patients were divided into restenosis group( n = 29) and no-restenosis group (n = 123 ). The serum LP (a) levels of all patients were also investigated. The general clinical data were analyzed. Multivariate logistic regression was used for statistical analysis. Results We compared the serum Lap (a) levels, smoking and diabet in the two groups, and there was a statisticaLly significant difference between the restenosis group and no-restenosis group(P<0.05). Multivariate logistic regression showed that the elevation of Lap (a) level re-mained as an independent predictor of restenosis (RR =2. 648,95% CI 1. 066-6. 575,P <0. 05). Other risk fac-tors,such as smoking(P =0.023) ,diabet(P =0. 036) and the type of stent(P = 0.011 ) were also correlated with restenosis. Conclusions High plasma LP (a) concentration is an independent predictor of stent restenosis after stent implantation.

Objective To investigate the molecular genetic characteristics and clinical characteristics of severe combined immunodeficiency(SCID)in children caused by a novel mutation of interleukin 2 receptor gamma IL-2RG gene.Methods The clinical data,laboratory results and genetic testing data of a child with SCID admitted to the Department of Children's Hematology of Northwest Women and Children's Hospital were analyzed.Results A two-month-old male infant was admitted to the hospital for treatment due to recurrent infections after birth.The child's blood routine results showed that the total number of white blood cells was normal,but lymphocytes were decreased.The lymphocyte subpopulation results showed a significant decrease in the proportion of total T(CD3+),helper T(CD3+CD4+),killer T(CD3+CD8+),and NK(CD3-CD16+CD56+)lymphocytes,while the proportion of B(CD3-CD19+)lymphocytes were increased.The immunoglobulin levels showed a significant decrease in IgG,and IgM and IgA were below the lower detection limit.The patient's cytokine levels did not significantly increase during infection.In the last three generations of the mother's family,9 males died of infection within one year after birth.The whole exome sequencing results of the core family revealed a semi zygous new missense mutation[c.675 C＞A,p.S225R(p.Ser225Arg)]in the IL-2RG gene on the X chromosome(chrX:70329160)of the patient,and the mother was a carrier.Based on the above evidence,the child was diagnosed with X-SCID.Subsequently,intravenous immunoglobulin was injected monthly,and routine antibiotics and antiviral drugs were taken to prevent infection,preparing for hematopoietic stem cell transplantation.Because the child was vaccinated with BCG after birth,the child developed disseminated BCG disease at the age of 6 months.After treatment,hematopoietic stem cell transplantation was performed.Conclusion The immune function of the X-SCID patient was severely compromised,which endangered the patient's life,and vaccination with live vaccines may lead to severe infections.This study found that the c.675 C＞A mutation of the IL-2RG gene was a novel pathogenic variation of the genetic cause of X-SCID,expanding the mutation spectrum of the X-SCID pathogenic gene IL-2RG.

Objective: To dynamically observe the progression of chronic gastritis to gastric cancer (GC) in disease-traditional Chinese medicine (TCM) pattern rats to provide data for understanding the disease pro-gression and effective approaches for drug screening and mechanism exploration.Methods: Wistar rats were randomly divided into control (n=96, half female and half male) and model (n = 336, half female and half male) groups. Model rats received free access to N-methyl-N′-nitro-N-nitrosoguanidine (120μg/mL), sodium deoxycholate (20 mmol/L), and alcohol (45％), and were subjected to intermittent fasting. Mortality rate, body weight, water consumption, food intake, gastric pathology, blood content analysis, and liver and kidney function of model rats were dynamically monitored over 30 weeks. In the 30th week, pattern characteristics were assessed. Gastric pathology and pattern charac-teristics were observed for an additional 8 weeks to evaluate stability. Results: The overall mortality of the model group was 34.82％(33.10％for females and 36.55％for males) at 30 weeks post-intervention. Inflammatory cell infiltration, glandular atrophy, atypical hyperplasia, and GC manifested successively in the gastric mucosa of rats. In model rats, N-methyl-N′-nitro-N-nitro-soguanidine intake was lower in males than in females, whereas pathological changes in the gastric mucosa occurred earlier in females than in males. Notably, gastric mucosal lesions were more severe in males than in females. Our modeling methods maintained stable gastric mucosal lesions for at least 8 weeks after final intervention. The pattern characteristics observed in model rats at the 30th and 38th week were consistent with those of spleen deficiency, blood stasis, and yin deficiency pattern. Blood content and indexes of liver and kidney function in the model group were normal. Conclusion: Our findings provide evidence for the pathological stages underscoring the progression of chronic gastritis to GC in disease-TCM pattern rats, which may facilitate development of relevant pharmacotherapies.

Objective To explore the evolvement rule of TCM syndrome types of the transformation of chronic gastritis (CG):from chronic non-atrophic gastritis (CNAG),chronic atrophic gastritis (CAG) to precancerous lesions of gastric carcinoma (PLGC).Methods Based on gastroscopy and gastrobiop-sy,altogether 592 patients included from three hospitals affiliated to Beijing University of Chinese Medi-cine Dongzhimen Hospital,(Dongfang Hospital and Third affiliated Hospital)and Wangjing hospital of China Academy of Chinese Medical Sciences from September 2013 to February 2014,were diagnosed with CNAG (338 cases),CAG (134 cases)and PLGC (120 cases).By using Epidata 3.1 the database was established,then principal component analysis,factor analysis and Chi-square test were conducted by using software SPSS 17.0.Results There were nine common syndrome types in CNAG,and the three leading ones were syndrome of incoordination between spleen and stomach,syndrome of heat-stag-nation in liver and stomach,syndrome of incoordination between liver and stomach.There were ten syn-drome types in CAG,and the highest frequency was syndrome of incoordination between liver and stom-ach,then syndrome of static blood in stomach collaterals and syndrome of deficient cold of spleen and stomach.The highest frequency of three syndrome of ten in PLGC were syndrome of yin deficiency in stomach,syndrome of yang deficiency in stomach and syndrome of static blood in stomach collaterals. The frequencies of location factors of CNAG from high to low were stomach,liver and spleen,the same as those of CAG;the frequencies of nature factors from high to low were qi stagnation,heat,qi deficiency, dampness and yang deficiency,which were some different from those of CAG being qi stagnation,heat, yang deficiency,dampness,blood stasis,yin deficiency and qi deficiency.With the disease progression, the frequencies of location factors of PLGC from high to low were stomach,spleen and liver,while those of nature factors were heat,yang deficiency,yin deficiency,qi stagnation,blood stasis,dampness,and qi deficiency.The distributions of location factors as well as nature factors were significantly different dur-ing the whole procedure of CG progression,CNAG,CAG and finally PLGC.Conclusion During the procedure of:CNAG turned into CAG and into PLGC in the end,the evolvement characteristic of “sthe-nia transforming into asthenia and appearing yin deficiency and blood stasis gradually”was found.

Objective To summarize the basic characteristics of fecal class Chinese medicines in modern clinical practice, and to find out the application of them in ancient prescriptions. Methods Taking“fe-ces”(Chinese:粪便;pinyin:fènbiàn) and “Chinese medicines”(Chinese: 中药; pinyin: zhōngyào) as the keywords, we searched CNKI, Wangfang, CQVIP and CBM databases of relevant articles pub-lished since the time of establishment. Then the fecal class Chinese medicines were screened. What we obtained were where the official names came from, what their alternative names were at different times, which creatures they were excreted from, what property and flavor, meridian tropism, efficacy and indi-cations they had. Furthermore, we searched the ancient prescriptions involving fecal class Chinese medi-cines using the database platform of Dictionary of Chinese Medicine Prescriptions, institute of Chinese Materia Medical of China Academy of Chinese Medical Sciences to get the common compatibility and po-tential rules of clinical application. Results Total eleven fecal class Chinese were common used in mod-ern clinical practice, which were recorded in various monographs of Chinese medicine by numerous names, and were excreted from birds, mammals and worm. The nature of the medicines were hot or cold;the flavor coved sour, bitter, sweet, bitter, salty; the meridian tropism lied in meridian of liver ( gall bladder) ,heart ( cardiac ) , spleen ( stomach ) , lung ( intestine ) and kidney ( bladder ); the various effects were listed as the following: resolving superficies, clearing heat, dispelling pathogenic wind and removing dampness, resolving stagnation, expelling blood stasisfor removing obstruction in collaterals, re-moving phlegm, relieving cough, calming endogenous wind, inducing resuscitation, removing toxicity, destroying parasites, relieve itching, eliminating necrotic tissues and promoting granulation. In addition, we extracted relevant 1 550 relevant prescriptions containing the eleven medicines which had the internal pharmaceutical compatibility and association rules. Conclusion The application of fecal class Chinese medicines has a long history. The further study and innovating application will be much of value of re-search and application in the intestinal microecology and the diagnosis and treatment of various diseases.