Objective To investigate the correlation of epidermal distribution of lamellar bodies and expression of ceramidase with skin barrier dysfunction in polymorphous light eruption.Methods Forty-seven patients with polymorphous light eruption and 40 healthy volunteers were recruited into this study.Noninvasive instruments were used to measure skin sebum content,transepidermal water loss(TEWL)and water content in stratum corneum in all of the subjects.Then,tissue specimens were obtained from the lesions at sunexposed sites in the patients and normal skin of the healthy volunteers.The ultrastructure and distribution of lamellar bodies were observed with transmission electron microscopy in five lesion and control specimens.Immunohistochemistry was performed to detect the expression of ceramidase in the tissue specimens.Results Compared with the normal skin from healthy volunteers,the lesions from patients showed decreased number of lamellar bodies in the granular layer and prick cell layer with a disorganized arrangement.Ceramidase was positively expressed in 20 lesion specimens and 36 normal control specimens,weakly expressed in 21 lesion specimens and 4 normal control specimens,and negative in 6 lesion specimens; there was a significant difference in the expression of ceramidase between the lesion specimens and normal control specimens(P ＜ 0.01).The lesions also showed high TEWL(34.2191 ± 12.70 vs.16.8350 ± 6.50,P ＜ 0.01),lower water content in stratum corneum(22.7319 ± 8.71 vs.29.4250 ± 5.08,P ＜ 0.01)and similar skin sebum content compared with the normal skin.Conclusions There is a disturbance in the synthesis of ceramide in patients with polymorphous light eruption,which may contribute to the impairment of skin barrier.

ObjectiveTo explore the effects of epidermal proteins and lamellar bodies on skin barrier in glucocorticoid-dependent dermatitis.MethodsTotally,60 patients with glucocorticoid-dependent dermatitis and 40 normal human controls were eligible for this study.A noninvasive method using TewameterTM was applied to determine transepidermal water loss (TEWL) value in these subjects.Tissue specimens were obtained from the lesions of 13 patients with glucocorticoid-dependent dermatitis and normal skin of 10 human controls.Subsequently,haematoxylin and eosin(HE) staining was performed to observe the histopathological changes,immunohistochemistry to detect the protein expressions of K6,K10,K14,K15,loricrin,filaggrin,involucrin in epidermis,and electron microscopy(EM) to estimate the density of lamellar bodies in tissue specimens.ResultsCompared with the normal controls,the patients displayed an elevated TEWL value (P ＜ 0.05),which suggested an impaired epidermal barrier.Histopathology of lesions revealed nonspecific inflammatorychanges withmarkeddifferencesbetweendifferentclinicaltypesofglucocorticoid-dependentdermatitis.Immunohistochemistry revealed an attenuated expression of K10,K14,loricrin,filaggrin,involucrin and abnormal expression of K15 in lesional epidermis compared with the normal epidermis (all P ＜ 0.05),hinting a suppression of epidermal differentiation and proliferation as well as an impairment of cornified envelope structure.The number and density of lamellar bodies were also reduced in lesional epidermis compared with the control epidermis.ConclusionsCompared with normal skin,the structure of skin barrier is impaired in lesions of glucocorticoid-dependent dermatitis,to restore skin barrier is essential for the treatment of this entity.

Objective To observe the effect of ultraviolet irradiation comparising 95％ ultraviolet A (UVA)and 5％ ultraviolet B(UVB)on the proliferation of human epidermal keratinocytes(HEKs),in hope to offer a basis for the construction of a photodamaged skin model induced by sunlight.Methods HEKs were isolated from foreskin tissue and cultured in vitro.After several passages,the HEKs were irradiated with different doses(0,2.5,5,10,20,30,40,60 J/cm2)of UV comprising 95％ UVA and 5％ UVB.Methyl thiazolyl tetrazolium(MTT)assay was used to evaluate cell viability after 24 hours of additional culture.SPSS 17.0 software was used to calculate the median lethal dose(LD50)of ultraviolet radiation in HEKs.Results The proliferation of HEKs was inhibited by 0,1.03％,6.60％,17.28％,31.28％,49.59％,59.67％ and 70.99％ respectively after irradiation with UV of 0,2.5,5,10,20,30,40 and 60 J/cm2.A significant inhibition of cell proliferation was observed in HEKs irradiated with UV at a dose of no lower than 10 J/cm2 compared with unirradiated HEKs(F =62.11,P ＜ 0.05).The LD50 of UV in HEKs was 31.31 J/cm2.Conclusions Aas the dose of UV irradiation increases,the proliferative activity of HEKs decreases,with the LD50 of UV being 31.31 J/cm2.

Objective To study the protective effect and its mechanism of anti-tumor necrosis factor-?antibody (TNF-? Ab) on lung injury after cardiopulmanary bypass(CPB).Methods Twenty-eight healthy rabbits were selected and randomly divided into four groups:group Ⅰ only received open chest operation;groups Ⅱ-Ⅳ underwent CPB.In the group Ⅳ,rabbit TNF-? Ab (2 400 pg/kg) was dropped into the intracheal tube before operation and just after releasing the aortic clamp.Saline was given to the group Ⅲ in- stead.Blood neutrophils count,TNF-?,MDA from the right and left atrium in the four groups were determined perioperatively.Water volume,TNF-? mRNA,TNF-? protein,apoptosis and pathomorphological changes were measured in the lung tissues.Results TNF- ? Ab can restrain leukocyte accumulation,reduce releasing of TNF-? and MDA in the lung.It can also reduce the occurrence of apop- tosis and attenuate pathomorphological changes in the lung tissue.However,it cannot reduce the secretion of TNF-? at the transcrip- tion level and protein level.Conclusion Intratracheal TNF-? Ab administration has markedly protective effect on lung injury after CPB.

Breast Neoplasms ; metabolism ; pathology ; surgery ; Carcinoma ; metabolism ; pathology ; surgery ; Diagnosis, Differential ; Female ; Fibroma ; metabolism ; pathology ; surgery ; Humans ; Keratins ; metabolism ; Middle Aged ; Vimentin ; metabolism

Objective To observe the clinical efficacy and safety of Chinese medicinal recipe Psoriasis No.1 Formula in treating blood-heat type of psoriasis. Methods Eighty patients with blood-heat syndrome were randomly divided into trial group and control group,40 cases in each group. The trial group was given oral use of the decoction of Psoriasis No . 1 Formula (mainly composed of Radix Rehmanniae, Rhizoma Imperatae, Cortex Moutan,Rhizoma Smilacis Glabrae,Radix Glycyrrhizae,Flos Sophorae,Radix Arnebiae seu Lithospermi, Radix Paeoniae Rubra, Folium Isatidis, Periostracum Cicadae, Radix Scutellariae, and Radix Angelicae Sinensis). The control group was given Compound Qingdai Capsules orally. The treatment for the two groups covered 8 weeks. Psoriasis Area Severity Index(PASI)scores,and contents of interleukin-17(IL-17)and tumor necrosis alpha (TNF-α)were observed before and after treatment. And clinical efficacy and adverse reactions were compared between the two groups after treatment. Results (1) PASI scores of the two groups were significantly lowered after treatment(P < 0.01 compared with those before treatment),and the trial group had stronger effect on decreasing PASI scores than the control group(P<0.05).(2)The total effective rate of the trial group was 87.5％,higher than the control group(67.5％),but the difference was insignificant(P > 0.05). (3)After treatment , contents of IL-17 and TNF-α of the two groups were markedly decreased(P < 0.01 compared with those before treatment),but the difference was insignificant between the two groups(P > 0.05). (4)No severe adverse effect was found in the two groups during the treatment. Conclusion Chinese medicinal recipe Psoriasis No.1 Formula is effective and safe in treating blood-heat type of psoriasis, and its effect is superior to that of Compound Qingdai Capsules.

Objective To investigate the characteristics of activation-induced cytidine deaminase (AID) expression level in de novo acute leukemia (AL) patients, chronic myeloid leukemia chronic phase (CML-CP), chronic myeloid leukemia blastic crisis (CML-BC) patients and leukemia cell lines. Methods The expression level of AID mRNA was measured in 89 cases of newly-diagnosed acute lymphoblastic leukemia (ALL) patients, 79 cases of de novo acute myeloid leukemia (AML) patients, 5 cases of CML-BC patients, 5 cases of CML-CP patients and leukemia cell lines NB4, THP-1, KG-1, Raji, K562 by real-time quantitative reverse transcriptase polymerase chain reaction (RT-PCR), bone marrow mononuclear cells of 16 normal healthy donors were used as the control group. Results The expression levels of AID mRNA in 89 cases of ALL and 79 cases of AML were 0.006-7 463.175 and 0.005-69.107, the median expression levels were 3.785 and 1.812, the expression level of AID mRNA in the normal control group was 0.146-4.707, and the median expression level was 1.483, respectively. The AID expression levels of ALL, B-ALL, Burkitt leukemia, M4 patients and Raji cells were significantly higher than those of the normal control group (all P <0.05). Nevertheless, the AID mRNA expression levels of M3 patients and NB4, KG-1 cells were lower than those of the normal control group (all P <0.05). Furthermore, the AID mRNA expression levels of K562 cell were strikingly higher than that of the CML-CP patients (P<0.001), so were those of CML-BC, chronic myeloid leukemia myeloid blast crisis (CML-MBC), chronic myeloid leukemia lymphoblastic blast crisis (CML-LBC) patients. Conclusion AID gene shows high expression level in B-ALL, Burkitt leukemia and M4, low expression level in M3 and KG-1 cells, and obvious high expression level in CML-BC.

OBJECTIVETo study the anti-cataract effect of gigantol combined with syringic acid and their action mechanism.

METHODH202-induced lens oxidative injury in vitro rat model was establish to observe the impact of gigantol combined with syringic acid on lens transparency under a dissecting microscope. D-galactose-induced cataract rat model was established to observe the impact of gigantol combined with syringic acid on lens transparency under a slit-lamp. UV spectrophotometry was adopted to detect the inhibitory activity of gigantol combined with syringic acid against AR. Molecular docking method was used to detect binding sites, binding types and pharmacophores of gigantol combined with syringic acid in prohibiting aldose reductase.

RESULTBoth in vitro and in vivo experiments showed a good anti-sugar cataract activity in the combination of gigantol and syringic acid and a better collaborative effect than single component-gigantol and syringic acid and positive control drug Catalin. Molecular docking and dynamic simulation showed their collaborative AR-inhibiting amino acid residue was Asn160 and the major acting force was Van der Waals' force, which formed common pharmacophores.

CONCLUSIONGigantol combined with syringic acid shows good anti-cataract, their action mechanism is reflected in their good collaborative inhibitory effect on AR.

Aldehyde Reductase ; antagonists & inhibitors ; Animals ; Bibenzyls ; Cataract ; drug therapy ; enzymology ; Drug Synergism ; Female ; Gallic Acid ; analogs & derivatives ; pharmacology ; Guaiacol ; analogs & derivatives ; pharmacology ; Humans ; In Vitro Techniques ; Lens, Crystalline ; drug effects ; enzymology ; Male ; Rats ; Rats, Wistar

Objective To study biological effect of recombinant human erythropoietin ( RhEPO) on the expression of oligodendrocyte in the neuron glia antigen 2 ( NG2 ) , Nogo receptor-interacting protein 1 (LINGO-1), myelin basic protein (MBP) and myelin associated glycoprotein (MAG), and to explore the protective mechanism of RhEPO for oligodendrocyte after cerebral infarction.Methods Experimental rats were randomly divided into the treatment group ( RhEPO at a dose of 3 000 U/kg) or saline control group.Both groups received intraperitoneal injection of RhEPO after cerebral ischemia in 30 min, 3 h, 6 h, 12 h and 24 h, which was administered daily for 7 days.The modified neurological severity score ( mNSS) and histology were analyzed, and immunohistochemistry was used to detect the protein expression of NG2, MAG, MBP and LINGO-1.Results The overall mNSS of RhEPO treatment group significantly decreased compared with the saline control group on the seventh day after cerebral infarction ( P<0.05 ).Such treatment effect was more obvious in the treatment group at 30 min and 3 h ( P<0.01).Compared with the saline control group, the numbers of NG2 positive cells increased in RhEPO treatment group.In contrast, the expression of LINGO-1 protein significantly decreased (P<0.05), with a dramatic decrease observed at 30 min and 3 h ( P<0.01).However, the expression of MBP protein decreased more significantly in saline control group, while the level of the MAG protein expression increased.The differences were statistically significant ( P<0.05), especially at 30 min (P<0.01).Conclusions After cerebral ischemia, RhEPO promotes the proliferation of NG2 positive cells, and inhibits the expression of LINGO-1 and MAG proteins.RhEPO improves the proliferation and differentiation of oligodendrocyte precursor cells, which in turn protects neuronal function, particularly at the early phase of ischemia.

OBJECTIVETo investigate the effects of caveolin-1 on the biologic behavior of laryngeal squamous cell carcinoma HEp2 cell line in vitro.

METHODSEukaryotic expression vector of human caveolin-1 gene was constructed and transfected into HEp2 cells by Lipofectamine. The clones stably overexpressing caveolin-1 were identified by real-time PCR and Western blotting. Cell proliferation viability was tested by MTT assay. Anchorage-independent growth was determined by assaying colony formation in soft agar. Flow cytometry was used to assess the cell cycle and apoptosis. The relative phosphorylation level of EGFR and ERK1/2 were detected by Western blotting. Localization of caveolin-1 and EGFR were studied by laser confocal laser scanning microscopy.

RESULTSThe expression vector of caveolin-1 was constructed and three clones stably overexpressing caveolin-1 were obtained. Comparing with the parental HEp2 cells, the transfected cells exhibited a slower growth rate and formed fewer colonies in soft agar. The results of FACS analysis revealed that overexpression of caveolin-1 resulted in the cell cycle arrest at G0/G1 phase and increased the apoptotic cell fraction. EGFR was found to colocalize with caveolin-1 in transfected cells by confocal laser scanning microscopy and Western blotting results showed that overexpression of caveolin-1 reduced the phosphorylation of EGFR and Erkl/2.

CONCLUSIONOverexpression of caveolin-1 suppresses the growth of HEp2 cells and induces apoptosis and inhibition of EGFR-MAPK signaling pathway may be involved in its mechanism.

Apoptosis ; Blotting, Western ; Carcinoma, Squamous Cell ; genetics ; metabolism ; pathology ; Caveolin 1 ; genetics ; metabolism ; physiology ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Cell Survival ; Flow Cytometry ; Genetic Vectors ; chemistry ; genetics ; Humans ; Laryngeal Neoplasms ; genetics ; metabolism ; pathology ; Lipids ; chemistry ; Microscopy, Confocal ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Phosphorylation ; Polymerase Chain Reaction ; methods ; Receptor, Epidermal Growth Factor ; metabolism ; Signal Transduction ; physiology ; Transfection ; methods