Objective To study the abnormal proliferation and differentiation of keratinocytes in psoriasis, the distribution of type I transglutaminase was detected and analysed in situ using immunohistochemistry assay. Methods Frozen sections of skin biopsy specimens taken from the skin lesions of unexposed area of psoriasis patients and from the skin of the same area of normal controls were incubated with mouse anti human transglutaminase monoclonal antibody, and detected by peroxidase labeled goat anti mouse antibody with AEC as substrate. Results In normal control, positive staining was only seen between keratinocytes of granular layer, but in psoriatic lesions, it was observed throughout the whole epidermis except the basal layer, the intensity was strongest in granular layer, and spread to spinous cell layer with declining intensity. There was a significant difference in the intensity between normal control and psoriasis (P

Objective To study the relationship between promyelocytic leukemia(PML) protein and apoptosis and their roles in the pathogenesis of psoriasis. Methods Immunohistochemical techniques were employed to study the expression of PML protein in the uninvolved skin, progressive plaque lesions and regressive lesions. Apoptosis was detected with terminal deoxynucleotidyl transferase-mediated dUTP-DIG nick end labeling(TUNEL). Results There was a significantly upregulated expression of PML protein in progressive plaque lesions, compared with that in uninvolved skin adjacent to the lesions and regressive lesions, and there was nearly no expression in normal skin; there was obvious apoptosis of keratinocytes in the epidermis with intensive PML protein staining. Conclusion The increased apoptosis of keratinocytes in the lesions of psoriasis induced by PML protein might be a homeostatic mechanism to the hyperplasia of keratinocytes.

Objective To construct eukaryotic cell expression vector of human frangible histone triad (FHIT) gene. Methods A 456 bp cDNA fragment was amplified from the total RNA of normal human thyroid tissue by RT PCR method and cloned into plasmid pcDNA3. The cloned insert was identified by double digestion of the recombinant plasmid with restriction enzymes Kpn Ⅰ and Bst XⅠ and sequenced by Sanger dideoxy mediated chain termination. The expression of FHIT gene was detected by immunocytochemical methods. Results The results showed that the cDNA fragment included 456 bp entire coding region. The recombinant eukaryotic cell expression vector of pcDNA3 FHIT was constructed, and the sequence of the insert was identical to the published sequence. MM96L cells transfected with the pcDNA3 FHIT plasmid expressed high level of Fhit protein in cytoplasm. Conclusion The recombinant plasmid pcDNA3 FHIT can provide a strong molecular tool for the studies of neoplasm pathogenesis.

Objective To detect the FHIT gene promoter methylation and protein expression in mycosis fungoides(MF).Methods Tissue specimens were collected from 48 patients with MF and 18 normal human controls.FHIT protein expression was determined by immunohistochemistry,and methylation status of FHIT gene by methylation-specific PCR.Results Abnormal methylation of FHIT gene was found in 26(54.2%)out of the 48 specimens.Thirty(63.5%)specimens of MF were negative for FHIT protein,which was observed in all the control specimens.The promoter methylation of FHIT was closely correlated with the protein expression of FHIT,but unrelated to the sex of,tumor staging or lymph node metastasis in patients with MF.Conclusion The FHIT gene promoter methylation may contribute to the inactivation and abnormal expression of FHIT protein in MF.

In order to explore the clinical effect of the glass fiber-reinforced composite resin post core for severe defect restoration of front teeth,sixty-two teeth were restored for forty-one patients with severe defect restoration of front teeth by glass fiber-reinforced composite resin post core plus alumina all-ceramic crown.Through 0.6-2.0 years follow up,all the prosthesis worked very well without breaking or falling off,no gum coloring except two posts fell off after the restoration at sixth month.The patients satisfied with the restoration.

Objective To investigate the mechanism of tazarotene in active psoriasis vulgaris. Methods HB-EGF mRNA in active psoriatic lesions before and 10 days after the treatment with tazarotene was detected by hybridization in situ. Results There was nearly no expression of HB-EGF mRNA in psoriatic lesions (9.1%); after the treatment with tazarotene, there was expression of HB-EGF not only in basal layer (95.5%), but also focal expression in suprabasal layers of epidermis (77.3%). Conclusion Tazarotene can inhibit proliferation and induce apoptosis of keratinocytes though upregulating expression of HB-EGF in psoriatic epidermis.

Objective To investigate the effects of tazarotene on the proliferation of cultured human keratinocytes and IFN-?-induced expression of HLA-DR in those cells.Methods Keratinocytes were cul-tured from normal human skin in vitro,and were treated with various concentrations of tazarotene(10 -5 ,10 -6 ,10 -7 mol/L).At24h,48h after treatment,the effects on cell proliferation were assessed by MTT method.The expression of HLA-DR was determined using immunocytochemistry techniques in cultured human ker-atinocytes incubated with tazarotene,IFN-?or both for24h.Results①The proliferation of keratinocytes was decreased when exposed to10 -7 -10 -5 of tazarotene as compared to non-exposed keratinocytes after24h and48h.Moreover,the effects on cell proliferation by tazarotene were dose-dependent;②There was rare expression of HLA-DR in normal human keratinocytes.③HLA-DR expression was inducible significantly with500u/mL of IFN-?,but failed to be induced with10 -6 mol/L of tazarotene,in keratinocytes at24h af-ter treatment.④After24h combined treatment of10 -7 -10 -5 mol/L of tazarotene and IFN-?,the induction of HLA-DR expression was significantly stronger,in a dose-dependent manner,than IFN-?alone(P

Objective To investigate the FECH gene mutation in a Chinese family with erythropoietic protoporphyria, to explore the relationship between gene mutation and clinical manifestations so as to estab-lish a basis for the genetic diagnosis and treatment of erythropoietic protoporphyria. Methods Clinical data on a Chinese family with typical EPP was collected. Peripheral blood was obtained from patients, unaffected individuals in the family and 50 unrelated human controls. Genomic DNA was extracted and PCR was per-formed to amplify the whole coding regions (exons 1 to 11) of FECH gene and their flanking intron sequences followed by direct sequencing to detect possible mutations. Results Based on clinical symptom and por-phyrin levels, a diagnosis of erythropoietic protoporphyria was made in 3 family members. DNA fragments of expected size were amplified by PCR. Gene sequencing revealed a heterozygons mutation (IVS1 + 1G >C) in intron 1 of FECH gene in the proband, his sister and father, but not in unaffected family members or unrelated human controls. Also, an IVS1-23C/T polymorphism associated with low expression alleles was observed in intron 1 of FECH gene of the proband, his sister and mother. Conclusions A novel mutation in the donor splice site of intron 1 of FECH gene is first reported in a Chinese family with EPP; this muta-tion may lead to a deficiency of FECH gene and serve as a molecular basis of development of erythropoietic protoporphyria.

Objective To investigate the alteration of cell proliferation and heparin-binding epidermal-growth-factor-like growth factor(HB-EGF)mRNA expression in cultured normal human keratinocytes after acitretin treatment.Methods After a 12-hour incubation with 0.1 and 1 ?mol/L acitretin in normal human keratinocytes,the proliferation potency of the cells was detected by MTT colorimetric assay and the expression of HB-EGF mRNA was examined by real-time quantitative reverse transcription polymerase chain reaction(RT-PCR).Results Both keratinocytes growth inhibition and HB-EGF mRNA expression was dose-dependently induced by acitretin.Treatment with 0.1 and 1 ?mol/L acitretin inhibited keratinocyte proliferation by 10.2% and 14.4%,and elevated HB-EGF mRNA by 3.2-and 7.1-fold,respectively.Conclusion Upregulated HB-EGF mRNA expression induced by acitretin in normal human keratinocytes may be involved in regulating keratinocyte growth inhibition after acitretin treatment.

Objective To investigate the role of heparin binding epidermal growth factor-like growth factor(HB-EGF)in active psoriasis vulgaris.Methods HB-EGF mRNA and protein were detected by hy-bridization in situ and immunohistochemistry in normal skin tissues,lesional and non-lesional psoriatic skin of progressive stage.Results In normal skin tissues,the stain of HB-EGF mRNA and protein was located in the basal layer of epidermis(100.00%)and there was only a little expression in the suprabasal layers(16.67%).Focal overexpression was found in the suprabasal layers of non-lesional and peri-lesional psoriat-ic skin(88.00%,80.00%respectively);however,there was no HB-EGF mRNA protein expression in the superabasal layers of the central part of psoriatic lesions(0),and nearly no expression in the basal layer(4.00%).Conclusion HB-EGF may play an important role in the pathogenesis of early psoriasis.