Objective To investigate the role of a cis-acting element,cell cycle genes homology region (CHR),in the transcriptional regulation of human NFBD1 gene (a nuclear factor with BRCT domain 1) by conducting a site-directed mutagenesis analysis on the element in human NFBD1 promoter region.Methods Wild type of NFBD1 promoter reporter,NFBD1-PS1-325,was used as template to make CHR mutant by using PCR based site-directed mutagenesis.Dual luciferase reporter assay was used to determine promoter reporter activity.Adiamycin treatment was employed to investigate the role of CHR in the transcriptional downregulation of NFBD1 after DNA damage.Results Site-directed mutagenesis of CHR caused a significant decrease in NFBD1 promoter activity,and also attenuated the transcriptional down-regulation of NFBD1 after DNA damage.Conclusion CHR element might be involved in both basic transcriptional regulation and transcriptional downregulation of NFBD1 after DNA damage.

Objective To evaluate the value of diffusion-weighted imaging (DWI)in diagnosing the acute centrum ovale infarction, and also to investigate the pathogenesis of the infarction. Methods All 58 patients underwent conventional MRI and DWI scanning after symptoms’ onset. DWI findings were compared to the findings of T_1WI and T_2WI. Results The sensitivity and specificity in diagnosing the ischemia stroke were 96.4% and 98.8% within 7 days after onset. Of all the cases, 62.1% were associated with the cerebral large-vessel disease and emboligenic heart disease. Only 36.2% had a classic lacunar syndrome but 69.0% had more frequently an abrupt onset of symptoms. Conclusion DWI is of high accuracy for diagnosing centrum ovale infarction and detecting early infarction lesions which are difficult to be displayed in conventional MRI, and very helpful in differentiating the acute from non-acute lesions; symptomatic centrum ovale infarction is suggested to be associated with large-vessel and heart disease which should be distinguished from the lacunar infarcts.

Objective　To explore the effect of hypoxia on the apoptosis of cultured human umbilical vein endothelial cells（HUVECs） and the role of vascular endothelial growth factor（VEGF） in inhibition of apoptosis． Methods　①Culture and identification of HUVECs．②Establishment of hypoxic model（0，12，24，48 h）in HUVECs．③Incubation of HUVECs with VEGF(0 ng, 100 ng) under hypoxic condition for 24 h. ④Detection of apoptosis of HUVECs with TUNEL method． Results　The percentages of apoptosis were different under different hypoxic conditions． The longer hypoxic time was，the higher apoptosis percentage was．VEGF reduced the apoptosis of HUVECs induced by hepoxia． Conclusion　Over-apoptosis EVCs in one of the important factors for the impairment of endothelial function. HEGF inhibits the apoptosis of HVCs and having a pretive function on them.

AIM To explore the effect of hypoxia on apoptosis in human umbilical vein endothelial cells(HUVECs) and the role of ecdysterone(EDS)in inhibition of apoptosis. METHODS ① Culture and identification of HUVECs;② Hypoxic model(0,12,24,48 h) in HUVECs was established. While HUVECs was incubated 24 h with EDS(100 mg?L -1 ) under hypoxic condition. Apoptosis in HUVECs was detected by TUNEL;③ HUVECs received EDS 100 mg?L -1 in normal and hypoxic condition. After 24 h, vascular endothelial growth factor (VEGF) was detected with immunohistochemical technique. RESULTS The apoptosis percentages are different under different hypoxic condition. The longer hypotic the time was, the higher the apoptosis percentage was. EDS reduced apoptosis of HUVECs. EDS could enhance expression of VEGF protein in cardiac myocytes of rat in normal and hypoxic condition. CONCLUSION The role of hypoxia in HUVECs apoptosis is more significant along with prolonging of hypoxic time. VEGF play an important role in protective effect of EDS on endothelial cell apoptosis.

OBJECTIVETo explore the effect of curcumin on TGF-β2 regulated peroxisome proliferater activated receptor y (PPAR-γ)/platelet derived growth factor β (PDGF-β) signaling pathway in lung fibroblasts of mice.

METHODSC57BL/6 mouse lung fibroblasts were in vitro cultured with TGF-β2, curcumin, or TGF-β2 plus curcumin. The cell proliferation was detected by cell growth counting in the blank control group, low, middle, and high dose curcumin groups (5, 25, 50 μmol/L), the TGF-β2 (10 ng/mL) group, TGF-β2 (10 ng/mL) plus curcumin (5, 25, 50 μmol/L) groups. mRNA expressions of PPAR-γ, platelet-derived growth factor receptor β (PDGFR-β), fibroblast growth factor R1 (FGFR1) were detected using reverse transcription PCR. Protein levels of PPAR-γ and collagen-1 were detected using Western blot and ELISA in the blank control group, the TGF-β2 group, the TGF-β2 (10 ng/mL) plus curcumin 50 μmol/L group.

RESULTSCompared with the blank control group, curcumin 50 μmol/L showed the most significant inhibition on cell proliferation at 48 h and 72 h. Compared with the TGF-β2 group, TGF-β2 (10 ng/mL) plus curcumin 50 mol/L also showed the most significant inhibition on cell proliferation at 48 h and 72 h. Compared with the blank control group, mRNA expressions of PPAR-γ and PDGF-β, as well as protein expression of PPAR-γ increased, the collagen-1 expression also increased in the TGF-β2 group (P < 0.05). Compared with the TGF-β2 group, mRNA expressions of PPAR-γ obviously increased in the TGF-β2 (10 ng/mL) plus curcumin 25 μmol/L group and the TGF-β2 (10 ng/mL) plus curcumin 50 μmol/L group, higher than that in the TGF-β2 (10 ng/mL) plus curcumin 5 [μmol/L group (P < 0.05). mRNA expressions of PPAR-γ was higher in the TGF-β2 (10 ng/mL) plus curcumin 50 μmol/L group than in the TGF-β2 (10 ng/mL) plus curcumin 25 μmol/L group (P < 0.05). mRNA expressions of PDGF-β was lower in TGF-β2 (10 ng/mL) plus curcumin groups than in the TGF-β2 group (P < 0.05). Besides, PDGF-β mRNA expressions were lower in the TGF-β2 (10 ng/mL) plus curcumin 50 μmol/L group than in the TGF-β2 (10 ng/mL) plus curcumin 5 μmol/L group and the TGF-β2 (10 ng/mL) plus curcumin 25 μmol/L group (P < 0.05). There was no statistical difference in FGFR1 mRNA expressions between the TGF-β2 group and 3 TGF-β2 plus curcumin groups (P > 0.05). Compared with the TGF-β2 group, PPAR-γ protein expressions increased and collagen-1 protein expressions decreased in the TGF-β2 (10 ng/mL) plus curcumin 50 μLmol/L group (P < 0.05, P < 0.01).

CONCLUSIONSCurcumin not only could inhibit TGF-β2 induced proliferation of lung fibroblasts, but also could inhibit the synthesis of collagens. These might be associated with up-regulating PPAR-γ expressions and down-regulating PDGF-β expressions. Therefore, curcumin might inhibit the occurrence and developing of lung fibrosis through blocking PPAR-γ/PDGF-β signaling pathway.

Animals ; Cell Proliferation ; Collagen ; Curcumin ; pharmacology ; Fibroblasts ; metabolism ; Lung ; drug effects ; metabolism ; Mice ; Mice, Inbred C57BL ; PPAR gamma ; metabolism ; RNA, Messenger ; Receptor, Platelet-Derived Growth Factor beta ; metabolism ; Signal Transduction ; Transforming Growth Factor beta ; Transforming Growth Factor beta2 ; metabolism

In recent years, there have been lots of progresses in the studies on red cell diseases in China, especially bone marrow failure diseases including immuno-related pancytopenia, aplastic anemia, myelodysplastic syndrome, and paroxymal nocturnal hemoglobinuria. Numerous laboratory experiments as well as clinical researches have been carried out by Chinese hematologists, which brought about much clearer pathogenesis, more rational diagnosis methods and more effective therapies for red cell diseases.
Anemia, Aplastic ; diagnosis ; epidemiology ; etiology ; metabolism ; China ; Hematologic Diseases ; diagnosis ; epidemiology ; etiology ; metabolism ; Hemoglobinuria, Paroxysmal ; diagnosis ; epidemiology ; etiology ; metabolism ; Humans ; Myelodysplastic Syndromes ; diagnosis ; epidemiology ; etiology ; metabolism ; Pancytopenia ; diagnosis ; epidemiology ; etiology ; metabolism

To investigate the bioactivity of Nocardia rubra Cell (NC), the mice were used to assay the toxicity, the effects on immune organs, phagocytes of peritoneal macrophage and the antitumor activity by perfusion of NC to the stomach of mice. Results indicated that NC could obviously stimulate in vitro the phagocytosis of peritoneal macrophage from mice, and remarkably inhibit the growth of S180 in mice, and its LD50 was more than 10 g/kg. In conclusion, NC had low toxicity, it could significantly enhance the organism immunologic function and had obvious antitumor effect and the anti-infection effect against a pathogenic microorganism.

0.99). DC: reproducibility was good for neutrophils, lymphocytes monocytes, eosinophils and basophils. Comparison of the results by instrument with manual for normal samples in morphology, the correlation was better for neutrophils, lymphocytes and eosinophils (r:0.968～0.983) ,good for monocytes(r=0.917), not good for basophils(r=0.659);The WBC scattergram would change and alarm flags would display when there are neutrophilic stab granulocytes, abnormal or atypical lymphocytes and immature cells in higher proportion.Conclusions The analyzer can be used to test blood cell parameters accurately and reliably. Its main performance indices accorded with the experimental requirements; The results were credible. It is necessary to check with microscopy for DC before reported when it were doubtted.

Objective To investigate the serological characteristics and molecular basis of the B (A)phenotype in ABO blood group and provide the data for clinical transfusion of individuals with B(A) phenotype.Methods The ABO group antigens on red cells of the proband,family members and donors were identified by monoclonal antibodies and the ABO antibodies in sera were detected by the standard A,B,O cells.The compatibility testing for the proband and donors was detected by salted test,polybrene test and antiglobulin test.The coding region of exon 6 to exon 7 in ABO gene was amplified by polymerase chain reaction(PCR) and the PCR products were sequenced.The haplotypes of proband were analyzed by cloning and sequencing.Results It was showed that both A and B antigens were detected on red cells of the proband and her two family members,and there was anti-A_1 antibody in their sera.The serological phenotype of the samples are identified as the A_2B.DNA sequencing showed 261 G/del,297A/G,526C/G,657C/T,700C/G,703G/A,796C/A,803G/C,930G/A heterozygotes in exon 6 to exon 7.It can be deduced that genotype in the proband is B(A)_(02)/O_(01).The genotypes of her mother and grandmother-in-law were B(A)_(02)/B_(101) and B(A)_(02)/O_(01),respectively.After cloning and sequencing,two alleles B(A)_(02) and O_(01) in proband was showed.B(A)_(02) has snigle nucleotide change(700 C>G),which resets replacement of proline with alanine at position 234.Two donors with phenotype A_2B were identified as genotype B(A)_(02)/O_(01) and A_(208)/B_(101),respectively.The results of crossmatch testing is in accordane between the proband and two donors and there was no clinical adverse reaction after transfusion.Conclusions 700C>G in α-1,3galactosyltransferase allele(B allde)can result in B(A)phenotype in individuals with the phenotype of A_2B.The donors in the transfusion for the individuals with B(A) phenotype should include individuals with A_2B phenotype.

Objective To establish a PCR sequencing-based typing (PCR-SBT) method for simultaneous genotyping of human platelet antigen HPA-1 to HPA-16w.Methods All DNA polymorphism sites of HPA-1 to HPA-16w were obtained from the immuno polymorphism database.The specific primers were designed using Primer Premier 5.0 software to amplify nucleotide acid fragments encompassing each HPA polymorphism site.The primer sequence and PCR condition were optimized to obtain specific and single amplification product.The PCR product was purified and then sequenced to determine the HPA genotypes.Two standard DNA samples were detected using the HPA PCR-SBT method to examine the accuracy d this method.Sixteen reference samples (including 6 interference samples with HPA gene mutations) provided by 14th platelet immunology workshop of international society of blood transfusion (ISBT) in 2008 were also tested by this home-brew HPA PCR-SBT method.Results Total eleven pairs of primers were designed to amplify and sequence the sixteen HPA systems.The HPA genotypes of two standard samples were 1aa/2aa/3ab/4aa/5ab/6aa/7aa/8aa/9aa/10aa/11aa/12aa/13aa/14aa/15aa/16aa and 1aa/ 2aa/3aa/4aa/5aa/6aa/7aa/8aa/9aa/10aa/11aa/12aa/13aa/14aa/15aa/16aa,respectively.The 256 HPA genotypes of 16 reference samples were clear.128 genotypes among them were completely accordance with the results provided by ISBT report.Conclusions The PCR-SBT assay combining high-throughput DNA sequencer established in the study provides a simple,rapid and accurate method for HPA-1 to HPA-16w systems genotyping.The assay is suitable for routine clinical HPA genotyping and shows a broad prospect in further applications.