Immune rejection is the leading cause of graft failure,and the main way for preventing corneal graft rejection is the application of immunosuppressive drugs.However,in the recent years,cellular therapy has been a new research hotspot for its targeted effect and fewer side effect.A lot of researches showed that Treg cells which areimportant in inducing and maintaining immunological tolerance could directly induce immune tolerance in cornealtransplantation.In recent years,dendritic cells also are found to have a dual role in the immune system,except as antigen presenting cells to induce immune response.Immature or immunosuppressive cytokine-expressing dendritic cells can induce immune tolerance.Mesenchymal stem cells which have multiple differentiation potential can exert anti-inflammatory effects on immune cells and effectively inhibit organ transplant rejection in vitro and in vivo.As another hotspot besides Tregs,myeloid-derived suppressor cells can inhibit the proliferation of a broad range of immune cells (T and B cells,NK cells,and macrophages),induce T cells apoptosis,and even induce Tregs.This review provides an update of these four kinds of cells on their effects and developments in cellular therapy for experimental corneal graft rejection.

China ; Humans ; Severe Acute Respiratory Syndrome ; prevention & control

China ; Humans ; Respiration, Artificial ; Severe Acute Respiratory Syndrome ; complications ; diagnosis ; therapy

China ; Communicable Disease Control ; Humans ; Public Health

Acquired Immunodeficiency Syndrome ; prevention & control ; therapy ; China ; Health Policy ; Humans

Objective To estimate the effect of ginsenoside Rb1 on the production and clearance of cyclobutane pyrimidine dimer (CPD) as well as on the expression of two nucleotide excision repair-associated proteins,xeroderma pigmentosum group C (XPC) and excision repair cross-complementing group 1 (ERCC1),by ultraviolet B (UVB)-irradiated murine epidermal cells and human HaCaT keratinocytes.Methods Totally,42 BALB/c mice were shaved on the back and divided into four groups: untreated group (n =6),UVB group irradiated with UVB only (n =12),low-dose and high-dose Rb1 group (both n =12) treated with Rb1 of 0.5 g/L and 2g/L (100 μl/cm2) respectively two hours before UVB irradiation.The dose of UVB in the animal experiment was 180 mJ/cm2.Half of the mice in each group were killed at 0.5 and 16 hours respectively after the irradiation,then,the back skin was resected and subjected to the determination of CPD levels in the epidermis by immunohistochemical SP method.Some cultured HaCaT cells were divided into several groups to be treated with different concentrations (5,20,50 mg/L) of Rb1 before or after different doses (15 and 30 mnJ/cm2) of UVB irradiation,and cells were collected at 0.5 and 12 hours after the irradiation.Subsequently,genomic DNA was extracted and CPD was detected by dot blot hybridization.Some HaCaT cells were cultured with or without the presence of Rb1 (50 mg/L) and irradiated with UVB (30 mJ/cm2),then,the cells were collected immediately or at 0.5,2,4 and 12 hours after the irradiation,and total protein was extracted and subjected to immunoblot analysis for the quantification of XPC and ERCC1 proteins.Results There was a high level of CPD in the epidermis of mice at 0.5 hour after the irradiation,with no significant differences between these groups (P ＞ 0.05).The number of CPD-positive cells per high power field (× 400) in the murine epidermis at 16 hours was statistically lower in the low-and high-dose Rb1 group than in the UVB group (32.1 ± 8.5 and 14.6 ± 4.1 vs.67.3 ± 11.2,both P ＜0.01).The CPD level in HaCaT cells was similar between these groups at 0.5 hour after UVB irradiation,but was markedly decreased at 12 hours in Rb1-treated groups.After UVB irradiation,the protein expressions of XPC and ERCC1 decreased with time in untreated HaCaT cells but increased with time in Rb1 (50 mg/L)-treated HaCaT cells.In detail,the XPC/glyceraldehyde-3-phosphate dehydrogenase (GAPDH) protein ratio in untreated HaCaT cells was 0.68 ± 0.11 immediately after the irradiation,significantly higher than that at 0.5 hour (0.47 ± 0.09,P＜0.05),2 hours (0.45 ± 0.08,P＜0.05),4 hours (0.37 ± 0.06,P＜0.01),and 12 hours (0.18 ± 0.03,P ＜0.01),and that in Rb1-treated HaCaT cells was 0.56 ± 0.07 immediately after the irradiation,compared to 0.48 ± 0.14 at 0.5 hour (P＞ 0.05),0.68 ± 0.15 at 2 hours (P＞ 0.05),0.97 ± 0.20 at 4 hours (P＜0.01),and 0.79 ± 0.12 at 12 hours (P ＜0.05).The ERCC1/GAPDH protein ratio in untreated HaCaT cells was 0.28 ± 0.03 immediately after the irradiation,higher than that at 0.5 hour (0.25 ± 0.03,P ＞ 0.05),2 hours (0.21 ± 0.02,P＜0.05),4 hours (0.14 ± 0.02,P＜0.01) and 12 hours (0.11 ± 0.01,P＜0.01),and that in Rb1-treated HaCaT cells was 0.27 ± 0.04 immediately after the irradiation,compared to 0.24 ± 0.04 at 0.5 hour (P＞ 0.05),0.29 ± 0.05 at 2 hours (P＞ 0.05),0.35 ± 0.05 at 4 hours (P＜0.05),0.39 ± 0.05 at 12 hours (P ＜0.01).Conclusions Ginsenoside Rb1 shows no obvious effect on the UVB-induced production of CPD,but markedly accelerates the clearance of CPD,which may be partly associated with the upregulation of XPC and ERCC1 protein expression.

Objective To investigate IL-1?mRNA expression and protein secretion after ultraviolet irradiation in human keratinocyte(KC)originated SCC12F cells.Methods Expression of IL-1?mRNA was detected by Northern blot and the secretion level of IL-1?protein was analyzed by ELISA.Results There was a spontaneous expression of IL-1?mRNA in SCC12F cells in culture system of normal KC,which was increased along with the culture time,and reached the highest expression level in120hours.The secretion level of IL-1?protein was increased in a time-and dose-dependent manner in SCC12F cells after ultraviolet B(UVB)irradiation.Conclusions SCC12F cells may express IL-1?mRNA spontaneously under regular KC media.The secretion level of IL-1?protein shows both time-and dose-effect after UVB irradiation.

?AlM:To investigate the relationship of susceptibility of primary angle- closure glaucoma with glutathione S-transferase T1 ( GSTT1 ) and M1 ( GSTM1 ) polymorphisms.
? METHODS: Totally, 300 cases were collected from primary angle-closure glaucoma patients and 300 health volunteers were served as control group. The observation group were divided into chronic and acute primary angle-closure glaucoma groups, then multiplex PCR technology was used to detect the genetic polymorphisms of GSTM1 and GSTT1.
?RESULTS:The distribution frequencies of GSTT1-null genotype were 54. 3%, while it was 54. 0% in the control group, statistically no significance between control group and observation group (χ2 = 0. 053, P > 0. 05 ) ; The frequency GSTT1 - null genotype in chronic group of primary angle-closure glaucoma was 54. 9%, while it was 48. 6% in the acute group of primary angle - closure glaucoma, statistically no significance between control group and acute group(χ2=0. 064, P>0. 05), and chronic group (χ2=0. 037, P>0. 05); The distribution frequencies of GSTM1-null genotype was 59. 0%, while it was 55. 7%in the control group, statistically no significance between control group and observation group (χ2=0. 013, P>0. 05);The distribution frequencies of GSTM1-null genotype was 62. 3% and 58. 1% in acute and chronic group of primary angle - closure glaucoma patients respectively. Acute group of primary angle - closure glaucoma has no significant compared with control group (χ2 = 0. 005, P>0. 05), Chronic group of primary angle-closure glaucoma had no significant compared with control group (χ2 = 0. 047, P>0. 05).
?CONCLUSlON:No statistic significance relationship is found between primary angle-closure glaucom patients and the null genotypes of GSTM1 and GSTT1 susceptibility.

Objective: To establish an HPLC method for determining the content of glucosamine hydrochloride and its prepara-tions. Methods:A Phenomenex Luna NH2 column was used, and the mobile phase was ammonium phosphate buffer (dissolving 3. 5 g dipotassium phosphate and 0. 25 ml ammonium hydroxide into 1 000 ml water, adjusting pH to 7. 5 with phosphoric acid) – acetoni-trile (25 ∶75). The flow rate was 1. 5 ml·min-1 and detection wavelength was 195 nm. The column temperature was 35℃ and the injection volume was 20 μl. Results:The linear range of glucosamine hydrochloride was 1. 6-16. 0 mg·ml-1(r=0. 999 9). The av-erage recovery of glucosamine hydrochloride tablets and capsules was 99. 3% and 99. 5%, respectively, and RSDs were 0. 3% ( n=9). Conclusion:The method is simple, accurate and reliable, and suitable for the quality control of glucosamine hydrochloride and its preparations.

Objective To investigate Expressions of CD1a and CD83 molecules in cervical lesions. Methods Immunohistochemical method was used to examine the expressions of CD1a and CD83 molecules in tissue samples from 30 patients with squamous cell carcinoma (SCC) of the cervix, 30 patients with cervical condyloma accuminatum (CA) and 30 patients with cervicitis. Results With the increase in the invasiveness of cervical lesions, there was a decline in the density of immature CD1a+ dendritic cells. The average number of immature CD1a+ dendritic cells per high power field (HPF) was 3.45 in cervicitis tissue, 2.89 in CA tissue, 2.41 in SCC tissue. On the contrast, a significant increase was observed in the density of mature CD83+ dendritic cells in CA tissue and SCC tissue compared with the cervicitis tissue (0.057 celIs/HPF and 0.039 celIs/HPF vs 0.019 celIs/HPF, both P < 0.05). The positivity rates of HPV 16/18 and HPV 6/11 were 56.67% and 3.30%, respectively, in cervical carcinoma tissue, 73.30% and 6.67%, respectively, in CA tissue, 3.30% and 0, respectively, in cervicitis tissue. Conclusions Compared with CA tissue, less mature dendritic cells were observed in cervical carcinoma tissue, demonstrating that the antigen presenting cells in carcinoma tissue are insufficient to mount an adequate immune response to prevent lesional invasion.