Thalassemia is inherited hemolytic anemia caused by globin production disorder,clinically known as beta thalassemia and alpha thalassemia resulted from beta and alpha chain synthesis disorder.Significant anemic clinical manifestations are observed in intermediate -level and severe -level patients with the 2 types of anemia, affecting greatly the growth and life quality of children patients who need blood transfusion to stay alive.The incidence rate of thalassemia is as high as 7% -20% in South China,so the prevention and standaridized treatment of thalasse-mia and the management over target population draw more attention.This paper is a review on the latest development of blood transfusion,chelation therapy,health education,birth intervention for thalassemia.

Objective To observe-the different effects of 2 doses recombinant human granulocyte colony-stimulating factors (rhG-CSF) on Nogo receptor(NgR) expression in the brain tissue of neonatal rats after hypoxic-ischemic brain damage(HIBD) at different times in order to reveal the neuroprotective effects of rhG-CSF.Methods Seven-day neonatal Sprague-Dawley(SD) rats were randomly divided into 4 groups by drawing method:sham operation group,model group,low-dose rhG-CSF group and high-dose rhG-CSF group,24 rats in each group.Then each group was divided into 4 subgroups (6 rats in each subgroup)and all rats were exterminated at different times after HIBD(1 d,3 d,7 d and 14 d).In the low-dose rhG-CSF group and high-dose rhG-CSF group,the rats were given daily doses of rhG-CSF 50 μg/kg,100 μg/kg respectively for 7 days by subcutaneous injection immediately after the molding(total 7 injections).In model group,rats received an injection of same amount of 9 g/L saline.In sham operation group,rats received no special treatment.Brain tissues of rats from each group were collected at different time points.The expressions of NgR protein and NgR mRNA in the left brain tissue were detected by immunohistochemistry and real-time fluorescent quantitative polymerase chain reaction (PCR).Results Immunohistochemistry:NgR proteins were constitutively expressed in the cerebral cortex in sham operation group at each time point;compared with sham operation group,the expressions of NgR in model group were increased markedly at each time point (135.67 ± 16.63,173.98 ± 17.82,234.00 ± 14.70,319.59 ± 25.22),and the differences were statistically significant(all P ＜ 0.01);compared with model group,the expressions of NgR in the cerebral cortex in low-dose rhG-CSF group (134.35 ± 8.89,109.04 ± 12.62,75.99 ± 13.39) and high-dose rhG-CSF group (81.38 ± 12.25,80.14 ± 10.50,72.58 ± 13.66) on the 3rd,7th,14th day were reduced significantly (all P ＜ 0.01).Compared with low-dose rhG-CSF group,the protein expressions of NgR in the high-does rhG-CSF group were decreased faster,and had the marked difference on the 3rd,7th day (P ＜ 0.05).Real-time fluorescent quantitative PCR:compared with the sham operation group,the expressions of NgR mRNA increased gradually in the cerebral cortex in the model group (1.34 ± 0.24,1.88 ± 0.27,2.88 ± 0.84,4.26 ± 0.86),the differences in NgR mRNA expression were statistically significant at different times(all P ＜ 0.05) ; compared with model group,the expressions of NgR mRNA in low-dose rhG-CSF group on the 7th (1.08 ± 0.30),14th day (0.93 ± O.26) and high-dose rhG-CSF group on the 3rd (0.61 ± 0.10),7th (0.56 ± 0.28),14th day (0.47 ± 0.12) were significantly different (all P ＜ 0.05).The expressions of low-dose group and high-dose group were reduced gradually.The NgR mRNA expression reduced more quickly in the high-dose group than in the low-dose rhG-CSF group and had substantial difference between two groups in 3 days (P ＜ 0.05).Conclusions The findings suggest that rhG-CSF intervention can reduce the expressions of NgR in the brain tissues of neonatal rats after HIBD,and low-dose rhG-CSF also has neuroprotective effect,but it could be weaker than high-dose rhG-CSF.

Objective To illustrate that tumor necrosis factor-α (TNF-αt) / nuclear factor of activated T cells pathway is the main function way that mesenchymal stem cells (MSC) inhibit proliferation of pulmonary vascular smooth muscle cells during treating pulmonary hypertension (PH).Methods MSC from human umbilical cord was used to treat PH rat models induced by monocrotaline.Rats were divided into the control group,the PH model group and the MSC group.The general conditions of the rats were observed.Haemodynamics was detected.Pathological sections and immunohistochemistry method were used to detect the lung structure and tissue changes.Changing conditions of TNF-αt/NFAT were detected.Results Compared with rats in the PH model group,the general conditions of the MSC group tended to be normal evidently:the right ventricular systolic pressure (RVSP) dropped [(30.37 ±3.13) mmHg vs.(47.90 ± 3.45) mmHg,1 mmHg =0.133 kPa],the aortic pressure (MAoP) increased [(115.03 ± 16.01) mmHg vs.(92.78 ± 16.28 mmHg)],the thickening condition of arterial intima-media was evidently relieved [(17.22 ±1.21)％ vs.(31.68 ±2.26)％],the plasma TNF-α level decreased obviously [(842 ±76) ng/L vs.(245 ±24)ng/L],and the lung tissue TNF-o level decreased (0.172 ±0.024 vs.0.248 ± 0.051),and all the differences were statistically significant (all P ＜ 0.05).The activation of pulmonary artery NFATc2 in the MSC treatment group was apparently inhibited.Conclusions MSC therapy may perform the treating effect in PH by inhibiting the over-proliferation of inflammation related pulmonary vascular smooth muscle cells via TNF-oα/NFAT pathway.

Objective Type A lamins are encoded by LMNA and a major component of the nuclear lamina, which have been suggested to play important roles in chromatin organization, transcription, DNA replication, and cell apoptosis. The aim of this study was to analyze the bioinformation of zebrafish lamins. Methods A phylogeny analysis was figured out with protein sequences of different species by Clustal X and MEGA 4?0 software. Then we compared the lamin protein sequences of different species with that of zebrafish by BLAST tool from NCBI. A figure of synteny analysis results was done with lamin sequence information of humans, murine and zebrafish cited from UCSC, Vega and Ensemble. Results The a?nalysis results showed that lmna, lmnb1, and lmnb2 genes of zebrafish are highly conservative and they may be homology of human LMNA, LMNB1 and LMNB2 genes. Conclusions Zebrafish lamins and human lamins have homologous sequence similarity, indicating that these two genes are orthologous genes.

Objective To investigate whether human umbilical vein endothelial cells as a feeder layer was capable of supporting the growth of embryonic stem cells in vitro.Methods Human umbilical vein endothelial cells were isolated and cultured and then prepared as feeder cells after 3 passages.Alkaline posphatase activity(AKP) staining,stem cell surface marker test and karyotypes were conducted in different periods of cell culture.The suspension of stem cells cultured after 20 passages on endothelial cells were inoculated to the legs of severe combined immunodeficiency(SCID) mice subcutabeously to test the teratoma formation.Results E14.1 embryonic stem cells retained good colonies when they were cultured on endothelial cells for 3 passages and 8 passages.In addition,they expressed SSEA-1,Oct-4,and a membrane alkaline phosphatase to a high extent at passages 3 and 8.Embryonic stem cells cultured for 15 passages stably retaind a normal karyotype.Embryonic stem cells cultured on endothelial cells for 20 passages were inoculated into the hind leg of SCID mouse,teratomas containing three embryonic layers were recovered six weeks later.Conclusion Human umbilical vein endothelial cells would support effectively embryonic stem cells expansion,and provide a clinically safe method to expand ES cells for futureclinical application.

AIM:To investigate the potential of hematopoietic stem cells(HSCs)derived from mouse embryonic stem cells(ESCs)to reconstruct hematopoiesis in vivo.METHODS:Using a three-step method,a mice embryonic stem cell line,E14.1 was induced into hematopoietic stem cells.The cell markers with CD34+/Sca-1+ were identified by flow cytometry analysis,then HSCs(1?109 cells/L)from third-step were injected into SCID mice for observing teratoma formation.To validate function of HSCs,colonogenic cell assay was conducted and the hematopoiesis in lethally irradiated mice was reconstituted.RESULTS:The method of three-step differentiation,combined to use more hematopoietic stimulating factor promoted the E14.1 cell differentiation into HSCs with highest percent of CD34+/Sca-1+ cells(as high as 58.64%?4.20%)with more CFU-E,CFU-GM and CFU-GEMM populations.The cells showed the character of hematopoietic progenitors by Wright-Giemsa staining.Positive selected CD34+/Sca-1+ cells by magnetic sorting from third-step differentiation were transplanted into 7 lethally irradiated female mice while predominant hematopoietic reconstitution were observed in 10 d after transplantation,with 71.4%(5/7)successful engraftment rate.Three recipients showed that the cell population of the peripheral blood leukocytes,red blood cells and hemoglobin approached to normal index at 40 d after transplantation,but followed relative slow renew in platelet count.Survival rate in transplant group was 43%,compared to 100% mortality in control mice.Karyotyping assays confirmed the female mice with XY.CONCLUSION:The three-step differentiation and the culture conditions described here support the differentiation of mouse ESCs into HSCs.HSCs derived from mouse ESCs can reconstruct hematopoiesis.

AIM: To explore inductive methods of hematopoietic stem cell (HSC) from embryonic stem cells (ESC) in vitro. METHODS: Using mice E14 line, the first step was the primary differentiation in which the ESC form embryoid bodies (EB) in methylcellulose-based cultures with SCF and VEGF. The second step involved the plating of cells originating from the EB into three different system of cultures containing SCF, VEGF,IL-3, IL-6 and EPO for HSC. And identifying HSC by flow cytometry analysis, colonogenic cells assay and Wright-Giemsa stain were also used. RESULTS: By two-step differentiating, it showed that HSC differentiated slowly in methylcellulose medium, percent age of CD34+/Sca-1+ cells slight increased about(31.5?4.7)% after day 14 induction. However, EBs were induced after 10 days to fast differentiate for HSC with more cells population by coculture on bone marrow stromal cells feeder. Flow cytometry analysis showed that percentages of CD34+/Sca-1+ cells might reached to (47.8?6.3)%. The more optimistic system of differentiation was bone marrow stromal cells feeder (BMSCF) in combination with supernatants of stromal cells from mice fetal liver(SSCFL), it significant supported differentiation of ESC into HSC with higher percent (53.6?7.2)%. Colonogenic cell assay and Wright-Giemsa stain confirmed that it possessed character of hematopoietic progenitors. CONCLUSION: Using methods of two-step differentiation, mice ESC were induced to differentiate into HSC by coculturing with BMSCF and SSCFL in combination of SCF,VEGF,IL-3,IL-6 and EPO.

Objective To explore the clinical and genetic characteristics of 17 cases with glucose-6-phosphate dehydrogenase(G6PD) deficiency in Guizhou,China.Methods The clinical features of 17 patients with G6PD deficiency were analyzed,DNA samples were obtained from the patients and some mothers,and the exons and flanking intronic sequences of the G6PD gene were analyzed by the polymerase chain reaction and sequencing.Results The cases had diverse phenotypes,these patients had acute haemolytic anaemia triggered by eating broad beans,infections,ingestion of specific drugs or the neonatal period and chronic nonspherocytic haemolytic anaemia.Three cases of the patients had concomitant diseases for α Mediterranean anemia,acute myeloid leukemia M2 type and neonatal anal membrane stenosis,respectively.G1376T,G1388A and A95G were the commonest G6PD variants in patients in Guizhou,China.G1376T,G1388A and A95G mutations were observed in 82.4％ cases.Two patients had only compound variants(c.1311 C ＞ T,IVS11 nt 93 T ＞ C).One case in the Rongjiang County,Guizhou Province had novel compound variants (c.G1388A,IVS10-10 T ＞ G) in the world.A patient's mother in the Guiyang City,Guizhou Province,China had compound variants (c.1376 G ＞ T,1311 C ＞ T,IVS11 nt 93 T ＞ C) as a carrier.Conclusions G6PD deficiency has a wide range of clinical heterogeneity.A novel G6PD compound variant haplotype c.G1388A,IVS10-10 T ＞ G was first found in the world,and the SNP spectrum of G6PD was enlarged.There may be a G6PD compound variant haplotype c.1376 G ＞ T,1311 C ＞ T,IVS11 nt 93 T ＞ C in Guizhou.

Objective To study the effects of marrow mesenchymal stem cells on heart functions and ventricular remodeling after myocardial infarction in rats. Methods Myocardial infarction model was established by ligation of the left anterior descending coronary artery (LAD) in adult SD rats. 4 and 8 weeks after MMSCs implantation, he-modynamic evaluations, left ventricular weight/body weight ratio and heart weight/body weight ratio were determined. HE staining was performed for counting microvasculars and Van Gieson staining was used for measurements and calculation of the myocardial fibrillar collagen. Then we investigated the migration and evolution of MSCs in vivo by fluorescent microscope. Results HR was significantly decreased in transplantation MMSCs group. Eight weeks after transplantation, body weight in transplantation MMSCs group reached that of control group. At the same time, SBP, DBP and MBP were significantly increased in transplantation MMSCs group. HR was significantly decreased in transplantation MMSCs group. Secondly, left ventricular weight/body weight ratio and heart weight/body weight ratio were significantly decreased 4 weeks after transplantation MMSCs. Then the ratio was significantly decreased 8 weeks after transplantation MMSCs. Thirdly density of microvasculars was increased at the boundary of infarction site in the animals transplanted MMSCs. Finally, total volume of the myocardial fibrillar collagen was reduced in the MMSCs treated groups after MI. Conclusion Transplanting MMSCs can improve the ventricular remodeling and heart functions in acute MI rat models.

Objective To study the effects of marrow mesenchymal stem cells on heart functions and ventricular remodeling after myocardial infarction in rats. Methods Myocardial infarction model was established by ligation of the left anterior descending coronary artery (LAD) in adult SD rats. 4 and 8 weeks after MMSCs implantation,hemodynamic evaluations,left ventricular weight/body weight ratio and heart weight/body weight ratio were determined. HE staining was performed for counting microvasculars and Van Gieson staining was used for measurements and calculation of the myocardial fibrillar collagen. Then we investigated the migration and evolution of MSCs in vivo by fluorescent microscope. Results HR was significantly decreased in transplantation MMSCs group. Eight weeks after transplantation,body weight in transplantation MMSCs group reached that of control group. At the same time,SBP,DBP and MBP were significantly increased in transplantation MMSCs group. HR was significantly decreased in transplantation MMSCs group. Secondly,left ventricular weight/body weight ratio and heart weight/bodyweight ratio were significantly decreased 4 weeks after transplantation MMSCs. Then the ratio was significantly decreased 8 weeks after transplantation MMSCs. Thirdly density of microvasculars was increased at the boundary of infarction site in the animals transplanted MMSCs. Finally,total volume of the myocardial fibrillar collagen was reduced in the MMSCs treated groups after MI. Conclusion Transplanting MMSCs can improve the ventricular remodeling and heart functions in acute MI rat models.