Objective To investigate the clinical significance of dynamic changes of CD4~+ CD25~+ regulatory T cells(Treg)in patients subject to allogeneic hematopoietic stem cell transplanta- tion(alIo-HSCT).Methods Forty-five patients received allo-HSCT.The graft-vesus-host disease (GVHD)was prevented by cyclosporine A and short-term MTX regimen in 31 patients.Fourteen of all the patients received Zenapox(CD25MAb)at the day of transplantation and day 4 after transplan- tation.The levels of Treg in peripheral blood were detected by flow cytometry from 45 patients at 2nd,4th,8th and 12th week after allo-HSCT and the time of aGVHD development,respectively.Re- suits Anti-CD25 could suppress the peripheral blood levels of Treg significantly.The Treg levels were significantly higher in patients with grade 0-1 aGVHD than those with 2-4 aGVHD at 8th and 12th week after transplantation.Among patients with 2-4 aGVHD,Treg levels were significantly low- er after development of aGVHD than before.Conclusions Treg are important for the aGVHD preven- tion and can be a useful clinical surveillant index for the development of aGVHD.It can significantly decrease the levels of Treg in the peripheral blood with anti-CD25.

Objectiive:The present study was designed to explore whether overexpression of human wild ?-synuclein in rat brain caused selective dopaminergic neuron loss in substantia nigra and aimed to find out a new method to make a rat model of Parkinson's disease(PD).Methods:The human wild ?-synuclein gene was induced into the rat brain by Adeno-Associated Virus(AAV) vector.The overexpression of ?-synuclein was detected by realtime PCR.The behavior of rats were recorded every 4 weeks after the viral particle injection.TH immunohistochemistry were performed at 4,8,12 and 16 weeks post-injection as well as the dopamine(DA),3,4-dihydroxypheny-lacetic acid(DOPAC) of striatum were determined by high performance liquid chromatography coupled with electrochemical detection.Results:Realtime PCR results revealed a significant overexpression of ?-synuclein in the injected hemisphere.By 8 weeks post injection,a significant loss of the dopaminergic neurons was observed.34% of the dopaminergic neurons were lost after 12 weeks,and about 60% cells loss after 16 weeks.The DA and DOPAC levels in the striatum decreased about 15% 12 weeks after injecting viral particle carried ?-synuclein gene and 30% decreased after 16 weeks.The AAV-?-synuclein-treated rats developed a type of motor impairment,i.e.,head position bias,compatible with this magnitude of nigrostriatal damage.Conclusion:All the results showed that overexpression of human wild ?-synuclein caused selective dopaminergic neuron loss and mimic a symptom of human PD in rats.This may be a new methed to make rat PD model which can offer new opportunities for the study of pathogenetic mechanismsand exploration of new therapeutic targets of particular relevance to human PD.

Objective To screen proteins binding with interferon?(IFN?)from human hepatic cDNA libraty by yeast-two hybrid technique.Methods The IFN?gene was amplified by polymerase chain reaction(PCR)and constructed into pGBKT7 vector as the bait plasmid in yeast-two hybrid system3,pGBKT7-IFN?was then transfected into yeast AH109.The transfected yeast were mated with yeast Y187 containing liver cDNA library plasmid in 2?YPDA medium.Diploid yeast was plated on synthetic dropout nutrient medium(SD/-Trp Leu-His-Ade)and synthetic dropout nutrient medium(SD/-Trp-Leu-His-Ade)containing X-?-gal for selecting.After plasmid extracting and en- zyme cutting analysis,the blue colonies were performed sequence analysis,the results were analyzed by bioinformatics.Results IFN?gene was successfully cloned and expressed in yeast cells.Thirty- four positive colonies were obtained using yeast-two hybrid technique.After sequence analysis,eight clones were found may have a binding effect with IFN protein.Conclusions IFN?genes was success- ful cloned and eight proteins that could bind with IFN?protein were also screened.

AIM:To observe the effect of fresh amniotic membrane transplantation ( FAMT) in acute ocular chemical burns.
●METHODS:A prospective study of 25 consecutive cases (36 eyes) with acute ocular chemical burns were treated with FAMT. The clinical efficacy was observed such as the time of amniotic membrane absorbed, corneal epithelialization & transparency, visual acuities and complications.
●RESULTS: With follow-up ranged from 3 to 6mo, 31 eyes′ amniotic membrane were dissolved in 2wk (86%). A total of 33 eyes showed corneal epithelialization in 4wk ( 92%) , 3 eyes showed persistent corneal epithelial defects and need secondary limbal stem cell transplantation or corneal transplantation ( 8%) . A total of 10 eyes showed superficial corneal vascularization (28%), 6 eyes′ cornea were opacity in part (17%), and one eye was symblepharon (3%).
●CONCLUSION:Early FAMT is an effective treatment in the management of acute ocular chemical burns to support epithelial healing, restore ocular surface integrity with potential to improve vision and reduce the incidence of complications. Furthermore, FAMT has advantages of easily obtain and convenient usage, which is suitable in local hospital of our country.

Objective: Cell cycle has recently become more appealing as a new target of anti-carcinogen-ic agent. Diallyl disulfide (DADS) inhibits growth and induces call cycle G_2/M arrest in human gastric cancer BGC823 cells. Cell division cycle protein 25C (Cdc25C) and CyclinB1 expression are involved in G_2/M arrest.However, mechanisms of G_2/M arrest are not yet fully understood. The aim of this study was to elucidate the mechanism of cell cycle G_2/M arrest in human gastric cancer BGC823 cells induced by DADS. Methods: The expression of chk1 and Chk2 mRNA associated with cell cycle arrest of BGC823 cells after the induction with DADS for 1 or 2 days was detected by RT-PCR. The protein expression of cycle-related proteins ATM-RAD3-related gene (ATR), checkpoint kinase1 (Chk1), checkpoint kinase 2 (Chk2), P-ATR, P-Chk1 and P-Chk2 was measured by Western blot. Interaction between Chk1/2 and Cdc25C was analyzed by immuno-precipitation. Results: After the cells were treated with 15 mg/L DADS for 1 or 2 days, the expression of Chk1 and Chk2 mRNA was not significantly different from that in untreated cells (P>0.05). Western blot analysis showed that the expression of total Chk1 and Chk2 treated with 15 mg/L DADS was not significantly different from that in untreated cells. But phospho-chk1 showed a significant increase after stimulation with 15 mg/L DADS for 2h to 12h and continued to increase gradually as time went on (P<0.05). Phospho-Chk2 showed a eak expression and a weaker expression after stimulation with DADS, but the changes were not statistically significant (P>0.05). Addition of 15 mg/L DADS to BGC823 cells for 15 rain to 120 min resulted in an increase in phospho-ATR expression, whereas no changes were found in ATR expression (P<0.05). The Chk1 Ab in-creasingly precipitated Cdc25C in BGC823 cells treated with DADS (P<0.05). In contrast, Chk2 Ab failed to change precipitation with Cdc25C by DADS (P>0.05). Conclusion: Activation of chk1 was involved in cell cy-cle G_2/M arrest in BGC823 cells treated with DADS. Cell cycle G_2/M arrest by DADS is associated with phos-phorylation of several cell cycle regulatory proteins including ATR and Chk1 which regulate expression of Cdc25C.

Hearing Loss, Sensorineural ; genetics ; Humans

Animals ; Brain Edema ; prevention & control ; Brain Ischemia ; physiopathology ; Erythropoietin ; pharmacology ; Female ; Hippocampus ; metabolism ; pathology ; Male ; Nitric Oxide ; metabolism ; Protective Agents ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; prevention & control ; Superoxide Dismutase ; metabolism

Aim To investigate the effects of diallyl di-sulfide( DADS) on G2/M arrest in Chk1/MGC803 and Chk2/MGC803 cells so as to establish stable human gastric cancer MGC803 cells with overexpression of Chk1/2 gene. Methods The colony formation, flow cytometry, RT-PCR and Western blot were used to de-tect the proliferation, cell cycle, and expression of Chk1/2 mRNA and protein, p-Chk1/2, CDC25C and cyclinB1, respectively. Results The colony formation showed that the colony forming efficiency in Chk1/MGC803 and Chk2/MGC803 cells treated by 30 mg· L-1 DADS was lower than in control group and vector group ( P <0. 05 ) . Flow cytometry demonstrated that 41. 3%, 57. 4%, 68. 9% and 42. 9% of G2/M cells in Chk1/MGC803 were increased than in MGC803 and Chk2/MGC803 , respectively after treated by DADS in 12,24, 36 and 48 h(P <0. 05). At the same time, RT-PCR disclosed that expression of Chk1 and Chk2 mRNA had no marked change. Western blot showed that total proteins of Chk1 and Chk2 and p-Chk2 had invisible change, but expression of p-Chk1 was up-reg-ulated, and CDC25C and cyclinB1 were down-regula-ted time-dependently in Chk1/MGC803 cells ( P <0. 05 ) . Conclusion DADS arrests MGC803 cells at G2/M by increasing p-Chk1 expression to cause down-regulation of CDC25C and cyclinB1 simultaneously.

Objective To investigate the efficacy of intratympanic methylprednisolone perfusion (IMP) for the treatment of sudden sensorineural hearing loss (SSNHL ) which failed to be fully responsive to conventional treatment .Methods The hearing outcomes of 87 patients with unilateral SSNHL were retrospectively analyzed .All of the patients received IMP after not fully responsive to conventional treatment of varying periods of time .They were hospitalized in our department between January 2008 and December 2012 and were followed up for at least one year to exclude recurrent hearing loss .Results The effective rate was 66 .67% and the mean PTA improvement was (18 .53 ± 13 .54)dB of the patients with the time interval between onset of symptoms and IMP within 15 days , 21 .21% and (5 .92 ± 15 .18)dB of the patients with the interval between 16 and 30 days ,4 .76% and (3 .69 ± 7 .00) dB of the patients with the interval more than 30 days respectively .The significant difference in the effective rates were compared among the three groups(χ2 =25 .91 ,P<0 .01) .Regarding to the PTA improvement ,the group with interval within 15 days was better than the other two groups(F=11 .182 ,P<0 .01) .A total of 30 cases acquired more than 15 dB hearing gain after IMP .One of them was hearing loss in low frequencies and the other 29 cases were hearing loss at all frequencies .The mean hearing gains of the 29 cases in 0 .25 kHz ,0 .5 kHz ,1 kHz ,2 kHz , 4 kHz and 8 kHz were 35 .17 ± 18 .15 dB ,35 .38 ± 15 .90 dB ,31 .28 ± 19 .74 dB ,21 .31 ± 17 .34 dB ,14 .97 ± 16 .00 dB and 13 .80 ± 16 .35 dB ,respectively .The mean hearing gains at lower three frequencies (0 .25~1 kHz)were better than those at higher three frequencies (2 k~8 kHz)(F=9 .494 ,P<0 .01) .Conclusion Receiving IMP earlier might help to acquire better hearing gain for the patients with SSNHL after not fully responsive to conventional treat‐ments .The hearing gains at the lower frequencies were better than those at the higher frequencies after IMP .

3T3-LI preadipocytes were induced to differentiate and 3T3-L1 adipocyte or preadipocytes were incubated with oleate or palmitate overnight.RT-PCR was used to measure adiponectin receptor(AdipoR)1 and AdipoR2 mRNA levels.The results showed that the AdipoRl and AdipoR2 expressions were differentiation- dependent.Oleate only suppressed AdipoR mRNA expression in preadipocyte but not in adipocyte.However,high concentration of palmitate reduced AdipoR mRNA expression in both 3T3-LI preadipocyte and adipocyte.