Objctive To explore the relationship between the expression of Fas/FasL and the apoptosis occurs in retinal ischemia/reperfusion injury of rats, as well as the therapeutic effects of bFGF on the ischemic retina. Methods The models of retinal ischemia/reperfusion injury was made by transient elevating introcular pressure. A total of 28 rats were divided into normal and operation group.The latter were subdivided into 1 hour, 6, 12, 24, 48 and 72 hours after reperfusion group, in which the left eyes of the rats were in the ischemia/reperfusion groups and the right ones were in the treatment groups (bFGF intracameral injection). Apoptosis was assessed by the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labelling (TUNEL) method, and the expression of Fas and Fas ligand was studied by strept avidin-biotin complex (SABC)immunohistochemistry. Results No positive cells were observed in the normal rats′ retinae, but there was a significant number of TUNEL positive cells in 6-24 hours after transient ischemia followed by a decrease at the 48th hour. The number of TUNEL positive cells reached a maximum at the 24th hour after ischemia. The expression of Fas gradually increased as early as when it was at the 6th hour, reached a peak at the 24th hour, and then decreased at the 48th hour. Similarly, the expression of Fas ligand was at peak in 24-48 hours in GCL and INL of retina. Conclusions Retinal ischemia-reperfusion after transient elevated IOP induced apoptosis of cells in the retina. Fas/FasL may play an important role in the early events of the apoptotic pathways. bFGF can rescue RGCs from retinal ischemia/reperfusion injury through downregulation of the expression of Fas/FasL and may represent an important mechanism for therapeutic neuroprotection.

Objective To explore the immunologic composition and T cell function of rhG-CSF mobilized peripheral blood grafts(G-PB) and rhG-CSF primed bone marrow grafts(G-BM) after being mixed in different proportions.Methods T cell subgroups,dendritic cells(DC) and its subsets in G-PB and G-BM were analyzed by multicolor flow cytometry.Immunologic composition of G-PB and G-BM after mixture in different proportions were calculated,lymphocyte proliferation ability and quantities of interleukin-4(IL-4),interferon-?(IFN-?) secretion by T cells in mixture grafts were determined using MTT and sandwich ELISA assays.Results After co-incubation of G-PB with G-BM in different proportions,T cell proliferation in mixture grafts was lower than that of NG-BM and G-PB(P

Aim To observe the changes on the expression of ?-SMA in rats with adriamycin-induced nephrothy and intervention of Valsartan. Methods The model group were made by unilateral nephrectomy with twice-adriamycin injections by caudal vein. Immunohistochemical method, flow cytometry and pathologic image analysis were used to observe the expression of ?-SMA, FN and LN in rats. Results The expression of ?-SMA was positive in the model group. Rates of positive cell and protein semi-quantitative analysis were more higher than the sham group(P

Objective To explore the difference of immunological characteristics between recombination human granulocyte colony-stimulating factor(rhG-CSF)mobilized peripheral blood grafts(G-PB)and steady-state bone marrow grafts(SS-BM).Methods From April to October 2003,G-PB and SS-BM of 15 related donors were collected.T cell subgroups,dendritic cells(DC),monocytes and the expression of CD28 costimulatory molecules on T cells were determined by multicolor flow cytometry.The lymphocyte proliferation ability and the quantities of interleukin-4(IL-4)and interferon-?(IFN-?)secreted by T cells were determined using MTT assays and sandwich ELISA.Results The absolute numbers of monocytes,CD3+,CD4+ and CD8+ T cells,and the ratios of CD4/CD8 in G-PB were significantly higher than those in SS-BM,respectively(P0.05).The quantities of IFN-? and IL-4 secreted by T cells per micromilter in G-PB was significantly higher than those in SS-BM(P0.05).The absolute numbers of DC1 and DC2 in G-PB were significantly higher than those in SS-BM(P0.05).Conclusion It is concluded that the difference of immunological characteristics between G-PB and SS-BM may explain the lower incidence of GVHD and lower relapse rate after SS-BM and G-PB transplantation.

Objective To investigate the role of immunophenotyping in distinguishing mycosis fun- goides (MF) from lichen planus and psoriasis.Methods The expression of CD1a,CD4,CD8,ICAM-1, LFA-1,HLA-DR,CD30 and CD7 was measured by ABC immunohistochemical technique in specimens ob- tained from lesional skin of 15 cases of MF,17 cases of lichen planus and 17 cases of psoriasis,and in the skin of 6 healthy controls.Results In the lesional epidermis of MF,the density of cells positive for CD1a, CD30 or ICAM-1,was significantly higher (mononuclear cells,P＜0.001;dendritic cells,P＜0.01) than that in the lesional epidermis of lichen planus,psoriasis and in the skin of healthy controls.The density of cells positive for CD4 or CD8 and of dendritic cells positive for HLA-DR was higher in lesional epidermis of MF than in that of lichen planus.The linear density of CD1a-positive cells (P＜0.01),the percentages of cells positive for ICAM-1 (P＜0.05) or LFA-1 (P＜0.05) were all higher in the lesional dermis of MF than in that of lichen planus.As far as the CD7-positive cell density was concerned,it was higher in the lesional dermis of lichen planus and psoriasis than in that of MF and skin of healthy controls (P＜0.01), while no difference was found between the epidermis of MF and that of lichen planus or psoriasis.Conclu- sion There are differences in the expression of CD1a,CD4,CD8,ICAM-1,LFA-1,HLA-DR,CD30 and CD7 in the lesional skin of MF,lichen planus and psoriasis,which may provide a clue to the pathogenesis of these diseases.

ObjectiveTo analyze the distribution and density of Langerhans cells and dermal CD68 positive histiocytes in lesions of secondary keloid.MethodsTissue specimens were resected from the lesions of 30 patients with secondary keloid and normal skin of 14 human controls.Immunohistochemistry was performed to observe the expressions of CD68 and CD1a in these specimens.A micrometer was used to count the number of positively stained cells per unit area.The Student's t test was conducted for data analysis by using the SPSS software.ResultsThe density of CD1a+ Langerhans cells was (61 ± 49) cells/mm2 in the epidermis of secondary keloid lesions, (258 ± 61 ) cells/mm2 in the control epidermis,and(40 ± 65) cells/mm2 in the dermis of keloid lesions.CD68+ cells were absent in the epidermis of keloid lesions.Significant differences were observed in the density of CD1a+ Langerhans cells between the lesional and normal control epidermis(t =9.88,P ＜ 0.001 ) and in the percentage of CD68+ cells in nucleated cells between the superficial dermis of lesions and control skin(62％ ± 12％ vs.70％ ± 14％,t =2.66,P ＜ 0.05).The density of dermal CD68+ histiocytes was similar between the lesions and control skin ((287 ± 73) cells/mm2 vs.(290 ± 22) cell/mm2,t =0.02,P ＞ 0.05).Conclusions In keloid lesions,Langerhans cells decrease in the epidermis but increase in the dermis,CD68+ histiocytes are absent in the epidermis,and reduced in the dermis with a declined percentage in nucleated cells.

Objecfive To study the effects of some cytokines such as TNF-α,IL-6 and IFN-γ as well as lipopolysaccharide on CD68 expression in HaCaT cells.Methods Human HaCaT keratinocytes were randomly divided into natural proliferation group (without stimulation),IFN-γ-stimulated group,TNF-α-stimulated group,LPS-stimulated group and IL-6 stimulated group.The work concentration of TNF-α,IL-6,IFN-γ and LPS was 50 mg/L.HaCaT cells were collected after 24-hour treatment with the cytokines followed by the examination of CD68 expression with flow cytometry,immunohistochemistry and reverse transcription(RT)-PCR,respectively.Results Compared with untreated HaCaT cells,the count of CD68-positive cells was elevated in cells stimulated by TNF-α(t=3.60,P<0.01),IL-6(t=3.93,P<0.01),IFN-γ(t=2.38,P<0.05)and LPS(t=2.52,P<0.05),and the effect of TNF-α and IL-6 was stronger than that of IFN-γ and LPS.Among the four cytokines,only IL-6 enhanced the mean fluorescence intensity of CD68-positive cells (t=8.34,P<0.01).After 24-hour treatment with TNF-α,IFN-γ and IL-6,CD68 expression was observed in the cytoplasm and on the membrane of HaCaT cells and was stronger in cells treated with TNF-α and IL-6 than in those with the other cytokines.A significant increase was observed in the CD68 mRNA expression after 24-hour treatment with TNF-α (t=4.34,P<0.01),IL-6 (t=7.52,P<0.01)and IFN-γ (t=2.81,P<0.05);TNF-α and IL-6showed a stronger promotive effect than IFN-γ.Conclusion IL-6,TNF-α,IFN-γ and LPS can upregulate the CD68 expression in HaCaT cells.

Objective To determine the incidence rate of HPV infection or multi-infections at different stages of cervical lesions in the development of cervical cancer , and the impact of specific types of HPV multi-infections on the risk of cervical cancer. Methods 103 samples of cervical tissues were detected and then divid-ed into ICC/HSIL group and LSIL/NILMF group according to the degree of pathological changes. HPV type was determined by PCR product sequencing. E6 nested multiplex PCR was performed to detect HPV multi-infections. Odds ratios were calculated to determinate the association between the sample category (LSIL/NILM or ICC/HSIL) and the specific types of HPV multi-infections. Results In HPV-positive samples, the rate of multi-in-fections had no significant differences between the two groups. Coinfection of HPV68 with HPV16 increased the risk of ICC/HSIL, as compared with HPV16 or HPV68 infection alone. Conclusions High-risk HPV coinfec-tions has a higher risk to induce ICC/HSIL than does HPV infection alone.

Objective To evaluate the effect of dexmedetomidine on the expression of brain-derived neurotrophic factor (BDNF) during lidocaine-induced spinal neurotoxicity in rats.Methods Fifty-six male Sprague-Dawley rats,weighing 250-280 g,aged 2-3 months,were divided into 7 groups (n =8 each) using a random number table:sham operation group (group S),lidocaine group (group L),normal saline group (group NS),3 different doses of dexmedetomidine groups (D1,D2 and D3 groups),and oα2-adrenoceptor antagonist yohimbine group (group Y).An epidural catheter was placed at L5.6 interspace.Ten percent lidocaine 20 μl was injected intrathecally in all groups except group S.Dexmedetomidine 5,15 and 25 μg/kg were injected intraperitoneally at 30 min before lidocaine injection in D1,D2 and D3 groups,respectively.In group Y,yohimbine 1.0 mg/kg was injected intraperitoneally at 15 min before dexmedetomidine 25 μg/kg was injected.Before intrathecal administration and at 24,48 and 72 h after intrathecal administration (T1-4),the Basso,Beattie,Bresnahan (BBB) Locomotor Rating Scale was used,and the tail flick latency to a thermal nociceptive stimulus (TFL) was measured to assess the locomotor function.The rats were sacrificed after the last behavioral test,and the lumbar segments (L3-5) of the spinal cord were removed for pathological examination and for determination of BDNF expression and cell apoptosis.The apoptosis index was calculated.Results Compared with group S,the BBB score was significantly decreased at T2-4,the TFL was prolonged,the BDNF expression was up-regulated,and apoptosis index was increased in L,NS,D1,D2 and D3 groups (P＜0.05).Compared with group L,the BBB score was significantly increased at T2-4,the TFL was shortened,the BDNF expression was up-regulated,and apoptosis index was decreased in group D3 (P＜0.05),and no significant change was found in the parameters mentioned above in NS,D1,D2 and Y groups (P＞0.05).Compared with group D3,the BBB score was significantly decreased at T2 4,the TFL was prolonged,the BDNF expression was down-regulated,and apoptosis index was increased in group Y (P＜0.05).The pathological changes of the spinal cord were significantly attenuated in group D3 as compared with group L,and there was no significant difference in pathological changes of the spinal cord between group Y and group L.Conclusion The mechanism by which dexmedetomidine reduces lidocaine-induced spinal neurotoxicity may be related to up-regulation of BDNF expression,and the mechanism by which dexmedetomidine up-regulates BDNF expression is completely related to activation of α2-adrenoceptors in rats.

Objective To compare the intensity and isotype of anticardiolipin(ACL)antibodies in the sera from patients with syphilis and SLE.Methods IgG and IgM type of ACL antibodies were examined by ELISA in99syphilis patients and75SLE patients.Results Although similar positive percentage of IgG ACL antibodies was shown in syphilis and SLE patients,stronger absorbance(A value)was seen in syphilis patients(P