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.

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.

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 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.

Adult ; Antibodies, Monoclonal, Murine-Derived ; therapeutic use ; Antigens, CD20 ; metabolism ; Antineoplastic Agents ; therapeutic use ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Cervical Intraepithelial Neoplasia ; drug therapy ; metabolism ; pathology ; surgery ; virology ; Cyclin-Dependent Kinase Inhibitor p16 ; metabolism ; Cyclophosphamide ; therapeutic use ; Doxorubicin ; therapeutic use ; Female ; Follow-Up Studies ; Humans ; Lymphoma, Follicular ; drug therapy ; metabolism ; pathology ; surgery ; virology ; Neoplasm Staging ; Neprilysin ; metabolism ; Papillomavirus Infections ; Prednisone ; therapeutic use ; Receptors, Complement 3d ; metabolism ; Rituximab ; Uterine Cervical Neoplasms ; drug therapy ; metabolism ; pathology ; surgery ; virology ; Vincristine ; therapeutic use

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

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.

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.

AIM　To study the apoptosis and characterizati on of cultured vascular smooth muscle cells (VSMCs) induced by cholestan-3β,5 α,6β-triol(Triol) and compared with 25-hydryoxycholestrol(25-OH). M ETHODS　The culture of vascular muscle smooth cells (VSMCs), light and e lectron transmission microscopy and TdT-mediated dUPT nick-end labeling (TUNEL ) technique. RESULTS　After being treated with oxyterols, VSMCs showed apoptosis of ultrastracture change including shrinkage, condensation of nuclear chromatin, fragmentation nuclei and formation of apoptotic body. TUNEL r evealed that Triol-induced apoptosis in VSMCs was in a concentration-dependent manner. The effect of Triol and 25-OH at a 30 μmol*L-1 concentration i n culture medium induced apoptosis of VSMCs, the former was not but the latter was inhibited by cholesterol at a 50 μmol*L-1 concentration. CO NCLUSION Triol can induce VSMCs apoptosis in vitro and oxysterol-i nduced apoptosis in VSMCs may be mediated through various pathway and different mechanism. Oxysterol-induced apoptosis in VSMCs may play an important role in t riggering atherosclerosis plaque rupture and result to the onset of the acute co ronary syndromes.