5.Recent advances of cellular therapy for corneal graft rejection
Chinese Journal of Experimental Ophthalmology 2017;35(9):848-851
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.
6.Expressions of CD1a and CD83 molecules in cervical lesions
Chinese Journal of Dermatology 2009;42(5):321-323
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.
7.Expression of hypoxia-inducible factor 1 alpha, vascular endothelial growth factor and angiopoie-tin-2 in tissues of condyloma acuminatum in pregnant women
Chinese Journal of Dermatology 2009;42(5):324-326
Objective To investigate the expression and significance of hypoxia-inducible factor 1 alpha (HIF- 1α), vascular endothelial growth factor (VEGF) and angiopoietin-2 (Ang-2) in tissues of condy- loma acuminatum (CA) in pregnant women. Methods Tissue specimens were obtained from the lesions of 30 pregnant women with CA and 30 non-pregnant women with CA, and from the vulva of 15 normal preg- nant women. By immunohistochemical staining, the expressions of HIF -1α, VEGF and Ang-2 were detected in these specimens. Results The expression rates of HIF -1α,VEGF and Ang-2 were 86.67%, 93.33% and 83.33% in pregnant women with CA, respectively, 63.33%, 66.67% and 53.33% in non-pregnant women with CA, respectively, and 0, 6.67% and 0 in normal women, respectively. Enhanced expressions of HIF -1α,VEGF and Ang-2 were observed in pregnant women with CA compared with the latter two groups (P < 0.05). In pregnant women with CA, a significant correlation was noted between the expression of HIF -1α and VEGF (r = 0.412, P < 0.01) and between the expression of Ang-2 and VEGF (r = 0.460, P < 0.01). Further more, the expression of HIF -1α, VEGF and Ang-2 positively correlated with each other in non-preg- nant women with CA. Conclusion In tissues of CA in pregnant women, HIF-1α,Ang-2 and VEGF are over expressed, which may be related to angiogenesis.
8.Interference by baicalin in the formation of photoproduct in epidermal cells from BALB/c mice irradiated with ultroviolet B
Chinese Journal of Dermatology 2009;42(2):132-134
Objective To investigate the formation and elimination of photoproduct in epidermal cells from BALB/c mice irradiated with ultroviolet B, and to observe the interference by baicalin in it. Methods BALB/c mice were randomized into 6 groups, I.e., blank control group receiving no exposure or protection, baicalin group receiving protection with baicalin, acetone group receiving acetone pretreatment, UVB group receiving UVB irradiation but no protection, UVB + baicalin group receiving UVB irradiation and protection with baicalin, UVB + acetone group receiving acetone pretreatment and UVB irradiation. Baicalin was applied at 1 mg/cm2 on the back of mice for 3 days in baicalin group and UVB + baicalin group. Twenty hours after the last application, UVB irradiation of 180 mJ/cm2 was given to mice in UVB group and UVB + baicalin group. Skin specimens were obtained from the tested sites at 1, 24, and 48 hours, respectively, after the irradiation. Cyclobutane pyrimidine dimers (CPD) was detected in the specimens with immunohistochemical staining and Southwestern dot blotting. Results CPD was observed only in irradiated mice. The relative content of CPD in epidermal cells 1, 24 and 48 hours after the irradiation was (100±5.22)%, (75.34±8.22)% and (42.11±3.24)%, respectively, in UVB group, (81.45±5.22)%, (32.14±6.33)% and ( 5.21±3.15 )% respectively, in UVB+baicalin group, ( 106±8.21 )%, (70.23±4.13 )% and (41.22±4.21)%, respectively, in UVB + acetone group. A significant difference was observed in the relative content of CPD between UVB group and UVB + baicalin group at 1, 24 and 48 hours after the irradiation (P<0.05, 0.01, 0.01, respectively). Conclusions Taken together, these results suggest that topical baicalin application mitigates DNA photo-damage. Baicalin is therefore a promising protective substance against UVB radiation.
9.Content Determination of Glucosamine Hydrochloride and its Preparations by HPLC
China Pharmacist 2016;19(2):387-389
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.
10.Protective effect of ginsenoside Rb1 on ultraviolet B-induced damage and its possible mechanisms
Chinese Journal of Dermatology 2013;46(7):496-500
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.