IGF-1 is involved in the growth and development of mammals,but its role in sebaceous gland excretion,B16 cell proliferation,and migration in the skin has not been reported yet.This study aims to reveal the function of IGF-1 by detecting its expression in animal skin.Using nano-body screening and purification methods,IGF-1 nanobodies(IGF-1-VHH)were obtained.Using IGF-1-VHH as the primary antibody,Western blot and immunohistochemistry were used to detect the expression of IGF-1 in bovine skin and alpaca acne.IGF-1-VHH was added to melanoma B16 cell culture medium,and CCK-8 and scratch healing methods were used to detect the effects of IGF-1 nanobodies on B16 cell proliferation and migration,as well as their possible molecular mech-anisms.The results showed that the obtained IGF-1-VHH could be applied in immunohistochemis-try and Western blot detection methods,with strong IGF-1 positive expression signals in bovine sebaceous glands and alpaca acne.IGF-1-VHH binds to IGF-1 in cells and has a certain inhibitory effect on the proliferation and migration of B16 cells by regulating the expression of RAS,ERK,and RAF.In summary,IGF-1 maybe involved in the secretion and excretion of sebum in animal skin.IGF-1-VHH inhibits the proliferation and migration of B16 cells by binding to IGF-1,provi-ding a new theoretical basis for ensuring normal physiological function of the skin and clinical di-agnosis and treatment of melanoma.

IGF-1 is involved in the growth and development of mammals,but its role in sebaceous gland excretion,B16 cell proliferation,and migration in the skin has not been reported yet.This study aims to reveal the function of IGF-1 by detecting its expression in animal skin.Using nano-body screening and purification methods,IGF-1 nanobodies(IGF-1-VHH)were obtained.Using IGF-1-VHH as the primary antibody,Western blot and immunohistochemistry were used to detect the expression of IGF-1 in bovine skin and alpaca acne.IGF-1-VHH was added to melanoma B16 cell culture medium,and CCK-8 and scratch healing methods were used to detect the effects of IGF-1 nanobodies on B16 cell proliferation and migration,as well as their possible molecular mech-anisms.The results showed that the obtained IGF-1-VHH could be applied in immunohistochemis-try and Western blot detection methods,with strong IGF-1 positive expression signals in bovine sebaceous glands and alpaca acne.IGF-1-VHH binds to IGF-1 in cells and has a certain inhibitory effect on the proliferation and migration of B16 cells by regulating the expression of RAS,ERK,and RAF.In summary,IGF-1 maybe involved in the secretion and excretion of sebum in animal skin.IGF-1-VHH inhibits the proliferation and migration of B16 cells by binding to IGF-1,provi-ding a new theoretical basis for ensuring normal physiological function of the skin and clinical di-agnosis and treatment of melanoma.

Objective To establish the HPLC fingerprint of Rosae Rugosae Flos;To provide references for the quality evaluation of Rosae Rugosae Flos.Methods The HPLC analysis was carried on a COSMOSIL 5C18-MS-Ⅱ column(4.6 mm×250 mm,5 μm);the mobile phase was 2.5 % acetonitrile + 0.1 % formic acid aqueous solution(A)-acetonitrile + 0.1 % formic acid(B)with gradient elution at the flow rate of 0.5 Ml/min;the column temperature was 40℃;the detection wavelength was 350 nm.The similarity of 13 batches of samples was evaluated by Similarity Evaluation System for Chromatographic Fingerprint of TCM(2012 edition).Qualitative analysis was carried out by LC-MS technology.The overall quality evaluation of Rosae Rugosae Flos was carried out by combining clustering analysis,principal component analysis and orthogonal partial least square discrimination.Results The common mode of HPLC fingerprints of Rosae Rugosae Flos was established,and the similarity of 13 samples was good.9 compounds were identified preliminary.13 batches of samples were aggregated into 3 categories by chemical pattern recognition.Conclusion The fingerprints of Rosae Rugosae Flos established in this study combines with chemical pattern recognition method,which has high sensitivity and strong specificity,can provide a basis for the quality evaluation of Rosae Rugosae Flos.

OBJECTIVETo investigate the effects of the histone deacetylase inhibitor, MS-275, on the immune molecule content and categories in hepatocarcinoma exosomes.

METHODSExosomes were isolated from the human hepatocarcinoma cell lines, HepG2 and Hep3b, and purified by a combination technique of ultrafiltration centrifugation and sucrose gradient ultracentrifugation. The expressions of heat shock protein (HSP)70, human leukocyte antigen (HLA)-I, HLA-DR, cluster of differentiation (CD) 80 and NY-ESO-1 on exosomes were analyzed with immunoelectron microscopy and Western blotting before and after MS-275 treatment. Intergroup differences were statistically analyzed by the Student's paired t-test.

RESULTSMS-275 treatment of both HepG2 and Hep3b cell types significantly increased the numbers of exosomes, their total protein content, and expression of HSP70, HLA-I and CD80 (per 100 exosomes), as compared to non-treated cells (all, P less than 0.01). MS-275 was also found to induce de novo expression of HLA-DR, but had no significant effect on NY-ESO-1 expression (P more than 0.05). The findings from immunoelectron microscopy confirmed those from Western blotting.

CONCLUSIONThe histone deacetylase inhibitor, MS-275, can significantly alter the immune molecule content and categories in exosomes of hepatocarcinoma cells. The differential expression profile may reflect an anti-cancer immune response and represent molecular targets for novel anti-hepatoma therapeutic or preventative strategies.

Antigens, Neoplasm ; immunology ; metabolism ; Benzamides ; pharmacology ; Carcinoma, Hepatocellular ; immunology ; metabolism ; Exosomes ; immunology ; metabolism ; Hep G2 Cells ; Histocompatibility Antigens Class I ; immunology ; metabolism ; Histone Deacetylase Inhibitors ; pharmacology ; Humans ; Pyridines ; pharmacology