Objective:The preventive/therapeutic effect via NF-κB activation epigenetic modulation induced by EGCG and co-effect on ALA-PDT by combined with EGCG in human A431 squamous cancer cells were investiga-ted. Methods:In A431 cancer cells induced by EGCG,the p16INK4ɑgene was detected by MS (methylation specific)-PCR. The modulated epigenetic effects in A431 cancer cells induced by EGCG,including antiproliferation effect de-tected by MTT assay. Expressions of NF-κBp65,DNMT1,HDAC1,CyclinD1,HIF-1α,VEGF,p16INK4ɑwere detec-ted by immunoblotting,and anti-metastatic effect displayed by cadherin-immunocytochemistry and the apoptosis in-duced by EGCG detected by Annexin V-FITC. The therapeutic co-effect of ALA-PDT combined with EGCG on A431 cancer cells apoptosis was detected by Annexin V-FITC. Results:The epigenetic pattern of A431 cancer cells could be modulated via demethylation of p16INK4ɑgene by EGCG. The down - regulated NF - κBp65,DNMT1, HDAC1,CyclinD1,HIF-1α,VEGF and upregulated p16INK4ɑinduced by EGCG were showed in immunoblotting. The anti-metastatic effect of enhanced cadherin -immunocytochemistry induced by EGCG also showed dose/time de-pendant. The apoptosis of A431 cancer cells could be induced by EGCG or ALA-PDT,further enhanced apoptosis of the cancer cells was induced by ALA-PDT combined with EGCG. Conclusion:EGCG with anti-oxidative activity can not only inhibit proliferative,vasculogenesis and metastatic effects via epigenetic-NF-κB signaling pathway to induce apoptosis of A431 cancer cells,but also can further promote the apoptosis induced by ALA-PDT combined with EGCG via pro-oxidant activity as cancer preventive/therapeutic agent.

Esophageal squamous cell carcinoma (ESCC) is the most common cancer in China, and multidrug resistance (MDR) remains one of the biggest problems in ESCC chemotherapy. In this study, we aimed to investigate the mechanism of Caveolin-1, an integral membrane protein, on regulating ESCC MDR. First, immunohistochemistry was used to check the protein expression of Caveolin-1, MDR-related protein of P-glycoprotein (P-gp), and multidrug resistance protein 1 (MRP1) in 84 pathologically characterized ESCC tissues, matched adjacent tumor, and adjacent normal-looking tissues. The results showed that Caveolin-1 expression level was elevated in ESCC tissues than that of matched adjacent tumor and adjacent normal-looking tissues (P < 0.05), and the expression of Caveolin-1 has close correlation with P-gp and MRP1 during tumor genesis of ESCC (P = 0.034, P = 0.009, respectively). Then, Caveolin-1 overexpression and knockdown were used to investigate its effect on expressions of P-gp and MRP1 in ESCC cell line Ec9706. The messenger RNA (mRNA) and protein expression levels of P-gp and MRP1 were checked by real-time quantitative reverse transcription-PCR (qRT-PCR) and Western blot (WB). The results showed that Caveolin-1 overexpression significantly promotes the mRNA and protein expression of MRP1 (P < 0.05), while almost has no effect on the mRNA and protein expression of P-gp (P > 0.05); Cavoelin-1 knockdown inhibits the mRNA and protein expressions of both P-gp and MRP1 (P < 0.05). The similar result was found in another ESCC cell line Eca109. So, it is concluded that Caveolin-1 affects ESCC MDR by regulating the expressions of P-gp and MRP1; therefore, it can be taken as a significant marker and target in tumor therapy.

Aim: To compare the expression levels of Nestin, NF-κBp65 and STAT3 in psoriatic lesions and normal samples. Methods: Immunohistochemical technique was used to determine the expressions of Nestin, NF-κBp65 and STAT3 in 14 psoriatic lesions and 14 normal samples. Results: The Nestin was located in the cytoplasm of respective stem cells in the basic cell layer, while that of NF-κBp65 or STAT3 was located in nuclei and cytoplasm in some basic layer cells and prickle layer cells. The expression levels of Nestin, NF-κB and STAT3 in psoriatic lesions were significantly higher than that in the normal samples [ Nestin(167. 51 ± 12. 93) vs (114. 38 ± 10. 85),NF-κBp65 (127. 48 ± 13. 87) vs (101. 27 ± 10. 15) and STAT3(196. 34 ± 10. 70) vs (124. 63 ± 19. 93), t = 12. 977,6. 288,and 13. 072,all P < 0. 001]. There was positive correlation between NF-κBp65 and STAT3 in psoriatic lesions and normal samples ( r = 0. 960 and 0. 829,P < 0. 001). Conclusion: The expression levels of Nestin,NF-κB and STAT3 are significantly higher in psoriatic lesion, and inhibiting their expressions may be valid manipulation for treatment of psoriasis vulgaris.

The cancer stem cell, CSC, also referred as cancer initiating cell, CIC, plays pivotal role in carcinogenesis, progression and metastasis. CSC/CIC possesses higher resistance to chemotherapy/radiotherapy and apoptosis than cancer cells, which can result in ineffective treatment and cancer relapse after treatment (Cetin and Topcul, 2012). Studies of CSC key markers principally aimed at selective elimination or establishment of therapeutic/ preventive target have become international hot spot in cancer research. CD133 is a universal marker used to identify and isolate normal stem cells (SC) or CSC from various organs or tumors (Tabu et al., 2013; Puglisi et al., 2009). A small subpopulation with stem cell properties, responsible for initiation of new tumor formation was selected by CD44+/CD24(-/low) marker in breast cancer (Carcia Bueno, 2008). A subpopulation of colon cells with CD44(hi+) has been identified to be highly tumorigenic as stem-like cells (Chu et al., 2009). CD44 is cell surface

Since the epigenetic alteration in tumor cells can be reversed by the dietary polyphenol quercetin (Q) or butyrate (B) with chemopreventive activity, suggesting that Q or B can be used for chemopreventive as well as therapeutic agent against tumors. In this study the polyphenol flavonoid quercetin (Q) or sodium butyrate (B) suppressed human esophageal 9706 cancer cell growth in dose dependent manner, and Q combined with B (Q+B) could further inhibit Eca9706 cell proliferation than that induced by Q or B alone, compared with untreated control group (C) in MTT assay. The reverse expressions of global DNMT1, NF-κBp65, HDAC1 and Cyclin D1 were down-regulated, while expressions of caspase-3 and p16INK4α were up-regulated, compared with the C group in immunoblotting; the down-regulated HDAC1-IR (-immunoreactivity) with nuclear translocation, and up-regulated E-cadherin-IR demonstrated in immunocytochemistry treated by Q or B, and Q+B also displayed further negatively and positively modulated effects compared with C group. The order of methylation specific (MS) PCR of p16INK4α: C>B/Q>Q+B group, while the order of E-cadherin expression level was contrary, Q+B>Q/B>C group. Thus, Q/B, especially Q+B display reverse effect targeting both altered DNA methylation and histone acetylation, acting as histone deacetylase inhibitor mediated via epigenetic-NF-κB cascade signaling.

CD133 was recently reported to be a cancer stem cell and prognostic marker. Quercetin is considered as a potential chemopreventive agent due to its involvement in suppression of oxidative stress, proliferation and metastasis. In this study, the expression of CD133/CD44 in esophageal carcinomas and Eca109/9706 cells was explored. In immunoflurorescence the locations of CD133+ and multidrug resistance 1 (MDR 1)+ in the same E-cancer cells were coincident, mainly in cytomembranes. In esophageal squamous cell carcinomas detected by double/single immunocytochemistry, small CD133+ cells were located in the basal layer of stratified squamous epithelium, determined as CSLC (cancer stem like cells); CD44+ surrounding the cells appeared in diffuse pattern, and the larger CD44+ (hi) cells were mainly located in the prickle cell layer of the epithelium, as progenitor cells. In E-cancer cells exposed to nanoliposomal quercetin (nLQ with cytomembrane permeability), down-regulation of NF-κBp65, histone deacetylase 1 (HDAC1) and cyclin D1 and up-regulation of caspase-3 were shown by immunoblotting, and attenuated HDAC1 with nuclear translocation and promoted E-cadherin expression were demonstrated by immunocytochemistry. In particular, enhanced E-cadherin expression reflected the reversed epithelial mesenchymal transition (EMT) capacity of nLQ, acting as cancer attenuator/preventive agent. nLQ acting as an HDAC inhibitor induced apoptotic cells detected by TUNEL assay mediated via HDAC-NF-κB signaling. Apoptotic effects of liposomal quercetin (LQ, with cytomembrane-philia) combined with CD133 antiserum were also detected by CD133 immunocytochemistry combined with TUNEL assay. The combination could induce greater apoptotic effects than nLQ induced alone, suggesting a novel anti-CSC treatment strategy.

肿瘤是目前危害人类健康的主要疾病之一。治疗肿瘤的主要手段包括手术、放疗、化疗和生物治疗等，但耐药现象的产生，导致药物治疗效果不甚理想。研究表明脑源性神经营养因子(BDNF)及其酪氨酸激酶爱体B(TrkB)在肿瘤的发生发展中起着非常重要的作用，激活BDNF/TrkB信号途径除了刺激肿瘤细胞存活，血管发生及失巢凋亡以外，还与肿瘤细胞的耐药性密切相关。本文主要综述BDNF/TrkB信号途径对肿瘤耐药的影响及相应的抗肿瘤治疗应用，以期为克服肿瘤多药耐药性提供新的研究思路，从而进一步提高肿瘤患者生存率和改善预后。

There is close relationship between neoplasia and inflammation. Many cancers arise from sites of chronic inflammation or irritation, and an oncogenic change always induces an inflammatory microenvironment to promote development of tumors. Inflammatory conditions in selected organs increase the risk of cancer, and in the tumor microenvironment smoldering inflammation contributes to proliferation and cancer progression. Natural bioactive compounds exhibit anti-inflammatory modulation (Pan et al., 2009; Gupta et al., 2011). The dietary flavonoid quercetin and resveratrol have nonsteroidal anti-inflammatory activity that may have applications for anti-inflammation treatment (Donnelly et al., 2004; Tuñón et al., 2009). Our other experiment indicates that the pro-inflammatory cytokine IP-10 in co-cultured human esophageal cancer 9706 cells can be inhibited by the polyphenol quercetin. Prevention of cancer has to be associated with prevention of inflammation, especially chronic inflammation. Flavonoid quercetin is one kind of yellowish powdered crystalline compound, (C15H10O7 in structure with five OH groups), widespread in vegetables, fruits and some

目的探讨纳米脂质体槲皮素（nanoliposomal quercetin，Q）下调角质形成细胞内核转录因子（NF—KB）和白介素一6（IL一6）表达的表观遗传学修饰效应，并与地塞米松（dexamethasone，D）的效应进行比较。方法将培养的HaCaT角质形成细胞分为三组：①IP-10＋Q组：20ng／mLIP-10＋40μmol／LQ；②IP-10＋D组：20ng／mLIP-10＋10^-7mol／I．D；⑧对照组（C组）：20ng／mLIP-10，三组细胞均培养48h。对各组细胞进行p16^INK4a的甲基化特异PCR（MSP），将各组细胞内提取的蛋白质进行DNA甲基化转移酶1（DN—MTI），组蛋白脱乙酰化酶1（HDACI），NF—κBp65，CyclinD1，IL-6及P16的免疫印迹并对各组细胞进行HDACl，IL-6免疫细胞化学染色，每组实验重复3次。对获得的数据进行相应的统计学处理。结果与C组相比，p16^INK4a的MSP显示：①组比②组p16^INK4a的DNA甲基化减低显著；免疫印迹显示：①组比②组的DNMT1，HDAC1，NF—KBp65，CyclinD1及IL-6的表达下调显著，p16^INK4a的表达上调显著；免疫细胞化学染色显示：①组比②组的IL-6及HDACI的表达下调显著。以上差异均有统计学意义（P〈0．05）。结论Q和D通过下调p16^INK4a甲基化和HDAC1的表观遗传-NF—κB信号途径而下调角质形成细胞增殖的NF—κB和IL-6炎症因子的表达，且Q比D的下调作用更显著。

目的探讨寻常性银屑病患者皮损中NF-κB,IL-6,p16的表达水平及各与DNMT1或HDAC1的共表达的关系。方法采用免疫组化法检测30例寻常性银屑病和30例包皮环切术皮损中NF-κB,IL-6,p16及各与DNMT1或HDAC1的表达和共表达,并利用计算机图像采集与分析系统对免疫组化结果进行平均光密度测定,分析各自表达水平及共表达定位的关系。结果寻常性银屑病皮损中NF-κB（96.25±1.92）和IL-6（129.11±10.47）的表达显著高于正常对照（NF-κB为67.53±2.03,IL-6为67.61±1.92）;p16的表达（64.92±1.97）显著低于正常对照（84.01±12.01）;此外,DNMT1和HDAC1各与NF-κB,IL-6,p16的共表达信号比较,正常对照组和病变组有差异,病变组在棘细胞浅层的表达均高于深层。以上差异均有统计学意义（P均＜0.05）。结论 NF-κB,IL-6,p16参与银屑病发病机制;棘细胞浅层可能参与银屑病的表观遗传调节。

目的探讨纳米脂质体槲皮素（nanoliposomal quercetin,nLQ）联合8-Br-cAMP对培养的人Rb44细胞组蛋白脱乙酰化酶的抑制效应。方法将培养的Rb44细胞分为4组:nLQ组,以40μmol·L-1nLQ培养;8-Br-cAMP组:以20μmol·L-18-Br-cAMP培养;nLQ+8-Br-cAMP组;以40μmol·L-1nLQ和20μmol·L-18-Br-cAMP共培养;对照组:不加nLQ或8-Br-cAMP。应用MTT实验检测细胞生长抑制率（GSR）。应用免疫斑点印迹检氉测脱乙酰化酶抑制物、NF-κBp65、磷酸化IκBα及c-myc的表达。对表达的靶信号以图像分析仪扫描灰度值。结果与对照组相比,nLQ组和8-Br-cAMP组GSR达44%±1%和40%±1%,均为P＜0.05;nLQ+8-Br-cAMP组GSR达60%±1%,P＜0.01。nLQ组、8-Br-cAMP组、nLQ+8-Br-cAMP组与对照组相比,脱乙酰化酶抑制物、NF-κBp65及c-myc的表达均减低,均为P＜0.05;而磷酸化IκBα的表达均增强,均为P＜0.05。结论 nLQ联合8-Br-cAMP可下调脱乙酰化酶抑制物表达,上调磷酸化IκBα的表达同时可抑制NF-κBp65的表达。

Objective: To study the effects of 8-Br-cAMP and quercetin alone or combined use on changes of mutator phenotypes in Eca-109 cells.Methods: The cultured Eca-109 cells randomized into 4 groups:8-Br-cAMP(Br) group,quercetin(Q) group,(Br+Q) group,and Control(C) group treated with no any drugs.After cultured 48 h,two kinds of specimens including cell slide and NCM specimens were prepared and Bcl-2-IR,Bax-IR,XRCC1-IR,MRP and POLB were detected by immunohistochemistry and the immunodot blot technique.Besides,the cell apoptosis was detected with TUNEL method in each group.Results: The cell apoptotic percentage was 70% in(Br+Q) group,42 % in Br group,35 % in Q group,and 4 % in C group.(Br+Q)Br or QC(P0.001).When compared with C group,Bcl-2-IR and XRCC1-IR were decreased and Bax-IR increased in the other 3 groups(P0.001),Br vs Q or Br vs(Q+Br)(P0.05).There waspositive correlation between Bcl-2-IR and XRCC1-IR,but negative correlated with Bax-IR(P0.01);The immunoreactivity(IR) showed:MRP-IR,CBr,Q or (Br+Q) group(P0.01);POLB-IR,CBr,Q or(Br+Q) group(P0.01).There was a signify cant positive correlation between MRP and POLB.Conclusion: The Br combined with Q or single Br or Q lone could down regulate the expression of Bcl-2,XRCC1,MRP and POLB and up-regulate that of Bax,(Br+Q) with highest cell apoptosis,which exhibites reduction of the mutator phenotype.

Aim:To explore cancer stem cell(CSC) markers in human esophageal squamous cell carcinoma(ESCC).Methods:Expressions of CD133,multi-drug resistance 1(MDR1)and CD44 were examined in 20 surgically resected ESCC tissues and Eca-109 and EC9706 cells by immuno-enzyme or immuno-fluorescence technique.Results:The percentage of CD133 and MDR1 positive cells wars 30%～43%,and the positive cells with CD133 and CD44 accounted for about 40% in Eca-109 and EC9706 cells.The small cells(CD133+) were located in the epithelial basal layer,cancer focus and at cancer pearl periphery.In ESCC roughly 40% CD44+ cells included small cell subpopulation located in the epithelial basal layer,cancer focus and at cancer pearl periphery and larger cell subpopulation mainly located in the epithelial prickle layer and scattered in the cancer focus.Conclusion:In ESCC cells,CD133 and CD44 may be the marker of CSC.

Objective To observe the TGase 1 and involucrin expression in nail of lamellar ichthyosis(LI) patients,and analysis the TGM sequencing.Methods The total protein was extracted from nails of 8 LI cases(LI group) and 8 healthy volunteers(C group),and performed by SDS-PAGE.The involucrin and TGase 1 were measured by Western blot and dot blot.TGM 1 gene sequencing was analyzed in blood samples from 2 LI cases.Results There were 5 major bands: 38 kDa,48 kDa,54 kDa 63 kDa and 69 kDa in LI group;4 former bands were demonstrated in C group.The involucrin blotting was mainly located at 63 kDa and 69 kDa bands in LI group,only located at 63 kDa major band in C group.The GSM of involucrin and TGase 1 in LI group compared with C group,P0.05.The missense point mutation was observed in the 8,12,13 exon in TGM 1 gene sequence of 2 cases of LI.Conclusions The TGM 1 gene sequences is abnormal,the involucrin is overexpression,the TGase 1 is lowexpression in nailof patients with LI.

Aim：To explore the mechanism for reverse transformation and apoptosis of Eca-9706 cells induced by nanoliposomal quercetin（nLQ）.Methods：A kind of liposomal quercetin, with chloroform＋DMSO being used as solvents,was prepared through rotary evaporation, and the nLQ was obtained after ultrasonic disruption and filtration.The cultured Eca-9706 cells were treated with nLQ at different concentrations,and cell growth inhibition rate for different exposure time （24,48,and 72 h） was detected by MTT.Eca-9706 cells were allocated into nLQ group （given 40 μmol/L nLQ） and control group.Apoptotic rate in the 2 groups was detected by TUNEL.After 48-hour-exposure,the expression of Cyclin D1,PTEN,c-Met,VEGF,HDAC1 and NF-κB in the 2 groups were detected by immunohistochemistry and immunoblotting.Results：Compared with the control group, the growth-inhibition rate （F24 h=128.041,F48 h=142.683 and F72 h=231.054,P0.001）and apoptotic rate（F=94.770,P0.001） of the groups of Eca-9706 cells had difference. The immunohistochemistry and immunoblotting showed that the expression of PTEN was up-regulated（t=5.352 and 4.308,P0.05）, while the immunohischemistry results of Cyclin D1, c-Met, VEGF,HDAC1 and NF-κB were down-regulated（t=4.006,9.184,13.853,4.698 and 3.575,P0.05）, and their immunoblotting results were also down-regulated（t=3.827,6.399,7.868,4.695 and 3.406,P0.05）.There was positive correlation in data above between immunocytochemistry and immunoblotting（rs=0.952,0.915,0.927,0.842,0.879,and 0.855,P0.05）.Conclusion：The Eca-9706 cell growth and proliferation could be inhibited by nLQ, and the overexpression of c-Met and VEGF and the low expression of PTEN in Eca-9706 cells could be reversed by nLQ. The Eca-9706 cells apoptosis could be induced through inhibiting HDAC1 and NF-κB and activating PTEN expressions.

Aim:To explore the expressions of the genes associated with growth and differentiation in peripheral blood leukocytes of lamellar ichthyosis (LI) patients.Methods:The peripheral blood leucocytes isolated from 8 LI patients and 8 healthy volunteers were subjected to detect the expressions of MAPK,CyclinD1,Filaggerin,Involucrin and K10 by immunocytochemistry and protein dot blotting,and those of c-fos,ras,c-myc,EGFR and iNOS by in situ hybridization and RNA dot blotting.Results:The gene expressions of c-fos,ras,c-myc,EGFR and iNOS,and expressions of MAPK,Filaggerin,Involucrin and K10 were attenuated in LI group than those in the control group(P0.01),while there was no difference in CyclinD1 expression between them(P0.05). Conclusion:The expressions of most genes associated with growth and differentiation in peripheral blood leukocytes of LI patients were down-regulated.

Objective To compare the difference in quercetin against oxidative stress response in mouse and in NIH-3T3 cells before and after H2O2 treatment,to explore the underlying mechanism for the quercetin antioxidant.Methods The cultured NIH-3T3 cells were randomly divided into 4 groups: quercetin(Q) pre-protective group(Qb) firstly treated with quercetin for 24 h followed by incubation with H2O2 for 30 min;post-protective group(Qa) treated with H2O2 for 30 min followed by incubation with quercetin for 24 h;H2O2 group(H2O2) after exposure to H2O2 for 30 min,incubated with DMEM medium and the control group(C) only cultured with DMEM medium.The survival rate and apoptotic rate were detected respectively with MTT and TUNEL in NIH-3T3 cell sus-pension samples.The expression of cyclin D1,PTEN,NF-κB,HSP-70,BCl-2,BAX and caspase-3 were examined with immunocytochemistry and immunoblotting.Besides,20 Wistar rats were divided into control group and experimental group,the latter was given with quercetin in the doze of 0.13 mmol/kg.The levels of T-AOC,SOD,GSH-Px,GSH,MDA,NOS and NO2-/NO3-were detected both in the cleaved NIH-3T3 cells and in the plasma from both experimental and control animals prior to and post-1 h,2 h and after 24 h.Results When the Qb group was compared with H2O2 or Qa group,the survival rate was higher and the apoptotic rate was lower.When the H2O2 group was compared with C group,the expression of cyclin D1、PTEN or BCl-2 was down-regulated;while that of BAX、HSP-70、NF-κB or caspase-3 was up-regulated;the level of T-AOC,SOD,GSH-Px or GSH was decreased;that of NOS、NO2-/NO3-or MDA enhanced in the cleft NIH-3T3 cells.When the plasma level of the anti-oxidative enzyme system prior to-compared with post-1h and 2h-treatment with Q,the level of T-AOC,SOD,GSH-Px and GSH,especially the former two,were higher;MDA,lower;NOS or NO2-/NO3-promoted.However,the above parameters basically became normal 24 h after treatment with Q.Conclusion Quercetin down-regulates the promoted expression of HSP70,NOS,NO2-/NO3-and NF-κB etc.in H2O2-treatment NIH-3T3 cells.Qb could reverse the H2O2 damage effects more markedly.Moreover,the quercetin exerts anti-oxidant protective effect through modulating the anti-oxidative enzyme system both in vivo and in vitro.However,based on the cell heterogeneity in none-or pre/post-H2O2-treatment state,a difference in quercetin antioxidant response is noted.

Objective To explore the relationship between the expression level and localization of CD80 and carcinoembryonic antigen(CEA) in the human fetal thymus and its development. Methods Twelve aborted dead fetus samples(via donation from the dead fetus's parents and permission of the hospital agency) were collected from Department of Genecology and Obstetrics in the affiliated hospital,including 4 cases aged less than 4 months in earlier stage and 8 cases aged 5 months in later stage. The two groups of earlier and later aged samples were randomly divided into two aliquots. One aliquot was prepared for frozen sections stained with HE,ANAE histochemistry,CD80 and carcinoembryonic antigen (CEA) immunohistochemistry; and detected with CEA ELISA. The protein extracted from another aliquot with Trizol reagent was detected with CD80 immunoblotting and CEA ELISA. Results The thymus in earlier aged group,especially the medulla was less developed. The reticulo-epithelial cells (RECs) with CD80-IR in the later aged group could be classified into six patterns. The type I RE in flatten shape were located at the inner surface of the capscule and trabecula. The type II RECs (DCs) in small satellite shaped were localized in the medulla. Both type III and type IV RECs were located at the dzemarcation between the cortex and medulla. The type V RECs (DCs) in large satellite shaped with delicate processes to connect into reticular meshes were distributed in the medulla and the type VI RECs were found in the Hassall's corpuscles. The ANAE-dot pattern T cells (CD4+) and ANAE-diffuse granular pattern T cells (CD8+) in separate clusters could be found in the reticular pores. In the immunoblotting and ELISA the expression level of both CD80-IR and CEA-IR in the later aged group was higher than that in the earlier aged group.Conclusion The results suggest that the expression and cocalization of CD80 and CEA closely related to the development and function of fetal thymus.

Background. The congenital lamellar ichthyosis (LI) patients display defective keratinocyte ( KC) envelops associated with TGM1 gene mutation, while analysis of them has been scarcely found. Objective. To explore the defective phenotype of KC involucrin and lipid envelopes detected in LI nail/scale samples. Methods. The trimmed nails and scraped scales were collected from 10 autosomal congenital ichthyosis (ACI) cases: five of them were recessive LI patients, the rest five patients were dominant ichthyosis vulgaris (IV) as patient control ( PC), and 10 healthy volunteers as the normal control (NC) group. The TGM 1 gene was sequenced in two individual LI patients from separate families. The extracted nail protein of the LI and IV cases was analyzed by SDS-PAGE and involucrin-immunoblotting. The involucrin immunostaining was performed on nail sections of extracted cross-linking protein and the scale spread slides; the histochemical lipid reactivity was examined on the scale spread slides. Results. In SDS-PAGE there were four major bands found in the NC group, while there were five major bands, including an additional major band of 69 kDa in the LI/IV patient group, especially distinct in LI cases. The involucrin expression was attenuated and the lipid reactivities, including phospholipid, cholesterol and unsaturated fatty acid in histochemistry, were decreased on the LI scale spread slides. The TGM1 gene sequencing showed that a frameshift mutation in Exon 3 and some point mutations in Exon 8, 12 and 13 were found in LI cases. Conclusion. The overexpression of 69 kDa and impaired envelopes may be potential biomarker associated with TGM1 problem for LI patients. [Life Science Journal. 2009; 6(1): 5 - 12] (ISSN: 1097 - 8135).

Objective To explore the effects of vitamin C and niacinamide on the growth and differentiation of human primary cultured keratinocytes.Methods Normal human foreskin was used in this study.The epidermis was separated enzymatically from the dermis by thermolysin,and keratinocytes were isolated from the epidermis by digestion with trypsin plus EDTA.The single keratinocytes were cultured with undedying NIH-3T3 cells as feeder cells in a complete medium supplied with 50 mg/L (vitamin C group),niacinamide of 400 μmol/L(niacinamide group)or vehicle(control group).Immunocytochemistry and immunodot blot were performed using monoclonal antibodies directed against C-myc,cyclin D1,filaggrin and involucrin.Results The colony number was highest in vitamin C group,followed by the control group and niacinamide group,and the colony morphology in vitamin C group was similar to that in the control group,but distinct from that in the niacinamide group.A significant increase was noticed in the expression of C-myc,cyclin D1,filaggrin and involucrin in vitamin C-treated keratinocytes compared with the control keratinocytes(all P＜0.05);however,in niacinamide-treated keratinocytes,the expression of filaggrin was significantly enhanced(P＜0.01),that of involucrin remained unchanged(P＞0.05),while that of C-myc was depressed(P＜0.05).Conclusions These results demonstrate that vitamin C has a favorable effect on both the growth and differentiation of human keratinocytes,while niacinamide seems to only promote the differentiation but attenuate the growth of human keratinocytes.