Clinical analysis of the infection with human papillomavirus in women / 中华实验和临床病毒学杂志

<p><b>BACKGROUND</b>To determine the relationship between human papillomavirus infection, cervical carcinoma, pre-cancerous lesion and condyloma acuminatum.</p><p><b>METHODS</b>From January 2004 to August 2005, 1086 inpatients in department of dermatology and department of gynaecology and obstetrics in Southwest Hospital and No. 302 Hospital with cervical lesions and condyloma were reviewed. All specimens were detected for HPV-DNA using techniques of Gene Array and fluorogenic quantitative polymerase chain reaction (FQ-PCR). All detections of HPV-DNA were performed in the first admission before the patients underwent any examination or treatment.</p><p><b>RESULTS</b>The positive rates of HPV-DNA detection were 100% in cervical intraepithelial neoplasia (CIN) I, and II and cervical carcinoma. Among these, the main subtype was HPV 16. But some of the patients were found to be positive for more than 2 subtypes of HPV. While the commonest HPV subtype was HPV 18 in endometrial cancer. Some of the patients were detected to be positive for more than 2 subtypes of HPV. In 636 female patient with condyloma acuminatum, the infection rates of HPV6, HPV11 accounted for 44.97% and 29.40%, respectively, HPV 16 and/or HPV 18 infection constituted a small percentage. In a few cases, infection with more than 2 subtypes was detected.</p><p><b>CONCLUSION</b>Cervical carcinoma including pre-cancerous lesion differs from condyloma acuminatum in dominate infectious subtype of HPV. The former is mainly associated with HPV 16 and HPV 18 infections, and the latter mainly associated with HPV 6 and HPV 11 infections. But in both of the above lesions, a mixed infection with more than 2 types may occur and make the pathological changes and clinical treatment more complicated. The early diagnosis and supervision of HPV infection may be of great value for improvement of prognosis and quality of life.</p>

Expressions of bFGF, ET-1 and SCF in dermal papilla cells and the relation to their biological properties / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the expression of bFGF, ET-1 and SCF in different passages of cultured dermal papilla cells (DPC), and their possible effect on biological behaviour of DPC.</p><p><b>METHODS</b>The expression of bFGF, ET-1 and SCF in different passages of cultured DPC was detected by immunocytochemistry and in situ hybridization.</p><p><b>RESULT</b>The expression of ET-1 and SCF in early passages of cultured DPC was stronger, but became negative in late passages (>6 passages). The stronger the expression of ET-1 and SCF in DPC, the higher ability of DPC to induce hair follicle regeneration.</p><p><b>CONCLUSION</b>The expression strength of ET-1 and SCF is related to the ability of DPC inducing hair follicle regeneration.</p>

Identification of differentially expressed genes in anagen dermal papilla by suppression subtractive hybridization / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>We constructed a cDNA subtractive library of dermal papilla cells (DPCs) in anagen with suppression subtractive hybridization (SSH) technique and clone differentially expressed genes related to DPCs in anagen.</p><p><b>METHODS</b>Total mRNA was isolated from DPCs of anagen and telogen follicles. Moreover, single-strand (ss) and double-strand (ds) cDNAs were synthesized in turn using SMART PCR cDNA synthesis technology. ds cDNAs then were digested with Rsa I and divided into two groups, and ligated to the specific adaptor 1 and adaptor 2R, respectively. After cDNAs were hybridized with each other twice and underwent two rounds of nested PCR. PCR products were ligated with arms of T/A plasmid vectors to set up the subtractive library. Selected clones were demonstrated by reverse Northern blot and sequenced. The acquired sequence data were aligned against the Genbank nucleotide database.</p><p><b>RESULTS</b>cDNA subtractive library of DPCs in anagen follicles was set up successfully with high subtractive efficiency. Thirty-five genes were identified in this study with 22 known functional genes and 13 unknown functional genes.</p><p><b>CONCLUSIONS</b>All results confirm the effectiveness and sensitivity of SSH in detecting differentially expressed genes from a small amount of clinical samples. Information about such alterations in gene expression could be useful for elucidating the genetic events in hair follicle growth regulation.</p>

Effects of cen-penetrating peptide on the presentation of MHC classⅠ-restricted cytotoxic T lympho-cyte epitope of human papillomavirus 16E7 / 中华皮肤科杂志

0.05).At the late stage (8-48 h) of incubation,the presence time of E7_(49-57)/K~b was significantly pro- longed on the surface of Tat-E7_(49-57)-incubated cells than that on the surface of other peptides-incubated cells (all P

The surveillance and molecular epidemiology of Aspergillus in the environment and patients at liver transplant department and intensive care units / 中华皮肤科杂志

Objective To explore the possible relationship between environmental contamination by Aspergillus and invasive aspergillosis.Methods From November 2005 to October 2006,samples were collected from the environment (air in corridors,air in wards,surfaces and tap water) twice a month,and from patients (nose,pharynx and sputum) at a liver transplantation department (LTD),neurologic surgery intensive care unit (NSICU) and central intensive care unit (CICU) in our hospital,and subjected to fungal culture.The Aspergillus density was determined in these environments.The isolates of Aspergillus flavus were genotyped by random amplification of polymorphic DNA (RAPD) assay to investigate the origin of infection.Results The mean aspergillus density was 12,10.75,0 and 20 cfu/m~3 at LTD,NSICU,CICU and corridors respectively.The five most prevalent species of aspergillus in these environments in decreasing order were Aspergillus flavus,Aspergillus fumigatus,Aspergillus niger,Aspergillus versicolor and Aspergillus clavatus.RAPD demonstrated that the genotypes ofA.flavus isolated from two patients were identical to those of the environmental strains in NSICU.The A.flavus genotypes from 3 patients in CICU were all different from those of the environment strains in CICU,but the genotypes were identical from two of the three patients.Conclusions Aspergillus contamination of different degree does exist at LTD,NSICU and CICU. The genotypes of A.flavus are identical from patients and environment in NSICU,suggesting that the clinical infection may originate from hospital environment.

A case of eumycetoma on the submaxilla due to Madurella / 中华皮肤科杂志

Objective To investigate the clinicopathological features,diagnosis,treatment,prognosis and causative agent of a case of eumycetoma on the submaxilla.Methods A case of eumycetoma diagnosed in our department was assessed for its clinical and pathological features as well as mycologic and molecular identification.Related literature was reviewed.Results The patient was primarily characterized by swelling of the submaxilla,with multiple sinuses draining many black granules.Pathologic examination revealed a pyogenic granulomatous inflammation,and a number of lotus rhizome node-like hypha were observed in tissue samples through PAS staining.Sequence analysis of multiple loci of the isolates,including ITS 1,ITS2 and D1/D2,showed that it was mostly similar to Madurella mycetomatis with a homology of 97%.Conclusion This is a case ofeumycetoma on the submaxilla induced by a novel species of Madurella.

Identification and the mRNA expression of HS1-associated protein X-1 in the peripheral blood mononuclear cells of patients with systemic lupus erythematosus / 中华风湿病学杂志

Objective To study the mRNA expression of HS1-associated protein X-1(Hax-1),an an- ti-apoptosis genc,in the peripheral blood mononuclear cells(PBMC)of patients with systemic lupus erythe- matosus(SLE),and further investigate the roles and significance of Hax-1 in the pathogenesis of SLE.Meth- ods Generation of longer cDNA fragments from serial analysis of gene expression(SAGE)tags for gene identi- fication(GLGI)was applied to identify the gene Hax-1 according to the Long SAGE tag.Then reverse tran- scription-polymerase chain reaction(RT-PCR)technique was used to semiquantitatively analyze mRNA ex- pressions of Hax-1 in PBMC from 34 active SLE patients and 25 healthy subjects.Results Compared with healthy controls,there was significant difference between SLE patients in the active stage and the normal controls(Z=-4.556,P＜0.01).The average level of mRNA expression in active SLE group was higher than that in healthy controls.Significant difference was found between the group with mild SLE and either the moderate or the severe one(P＜0.01).Conclusion The mRNA expression level of Hax-1 in active SLE group increase markedly,and to some extent,it is related to the activity of SLE.This provides a valuable basis for the further study on the role of apoptosis in SLE.

Factors related to collagen gel contraction in hair follicle organotypic culture / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the effects of rat tail collagen, hair follicle dermal papilla cells and hair follicle epithelium cells on collagen gel contraction in organotypic culture.</p><p><b>METHODS</b>The hair follicle organotypic culture was prepared with different concentrations of rat tail collagen, different number of dermal papilla cells and hair follicle epithelium cells in DMEM medium, after cultured for 10 days the diameter of collagen gel was measured.</p><p><b>RESULT</b>The concentration of rat tail collagen, hair follicle dermal papilla cells and hair follicle epithelium cells significantly influenced on collagen gel contraction in organotypic culture (P<0.01). The contraction of collagen gel was negatively related to the concentration of rat tail collagen, while the concentration of dermal papilla cells and hair follicle epithelium cells was positively related to the contraction of collagen gel.</p><p><b>CONCLUSION</b>The key factor influencing collagen gel contraction in organotypic culture is the concentration of rat tail collagen, hair follicle dermal papilla cells and hair follicle epithelium cells.</p>

Clinicopathological analysis on angiolymphoid hyperplasia with eosinophilia / 第三军医大学学报

Objective　To analyze the clinicopathological features of angiolymphoid hyperplasia with eosinophilia (ALHE). Methods　The pathological specimens of 7 cases of ALHE collected in our department from 1950 to 1999 were sectioned, stained and observed. Results　There were 3 pathological characteristics in ALHE: ①massive hyperplasia of capillaries in the dermis; ②the endothelial cells proliferated and swelled, projecting into vascular cavity like tombstones; ③mixed infiltration of lymphocytes and eosinocytes in the vessels. Conclusion　ALHE is a disease with local benign proliferated vessels, whose etiology and pathogenesis is still unknown. It is necessary to grasp the pathological changes of ALHE to distinguish it from other diseases.

Influence on the adhesion and growth of dermal papilla cells by chondroitin sulfate and heparin sulfate / 第三军医大学学报

Objective　To investigate the actions of extra cellular medium in growth and differentiation of hair follicle and to look for growth adjusting factors for dermal papilla cells (DPC). Methods　Dermal papilla cells were isolated and cultivated with two steps method and the cells were identified by immunohistochemical staining for actin. Influence was examined on the adhesion and growth of dermal papilla cells by chondroitin sulfate A, chondroitin sulfate C and heparin sulfate. Results　Two steps method of enzyme digestion for isolating and cultivating dermal papilla cells was an efficient method and large amount of dermal papilla of high purity were harvested with this method. The method is very simple and easy to manege with. Increased adhesion and growth of dermal papilla cells were observed in specimen treated with chondroitin A and heparin sulfate. No significant effects was observed in the cells treated with chondroit in sulfate C. Conclusion　Some extra cellular medium can regulate the adhesion and growth of dermal papilla cells and therefore influence the growth and development of hair follicle.

Experimental study of cytotoxic T lymphocyte associated molecule-4Ig for prevention of mice autoimmune hepatitis / 第三军医大学学报

Objective　To investigate the role of cytotoxic T lymphocyte associated molecule-4Ig(CTLA-4Ig) in the prevention of C57BL/6 mice autoimmune hepatitis. Methods　The C57BL/6 mice were intraperitoneally immunized with C57BL/6 mice liver-specific protein in complete Freund's adjuvant. At the same time CTLA-4Ig were given to observe the pathologic alteration of C57BL/6 mice liver. Results　With the increase of time of immunization, the results in the treatment group were similar to those of the control group; but inflammatory cell infiltration, hepatic cell swelling, focal necrosis and severe hepatocyte damage were found in the pathologic model group. There was a significant difference between the pathologic model group and control one. Conclusion　Autoimmune hepatitis of C57BL/6 mice can be effectively prevented by CTLA-4Ig.

Clinicopathological analysis on angiolymphoid hyperplasia with eosinophilia / 第三军医大学学报

Objective　To analyze the clinicopathological features of angiolymphoid hyperplasia with eosinophilia (ALHE). Methods　The pathological specimens of 7 cases of ALHE collected in our department from 1950 to 1999 were sectioned, stained and observed. Results　There were 3 pathological characteristics in ALHE: ①massive hyperplasia of capillaries in the dermis; ②the endothelial cells proliferated and swelled, projecting into vascular cavity like tombstones; ③mixed infiltration of lymphocytes and eosinocytes in the vessels. Conclusion　ALHE is a disease with local benign proliferated vessels, whose etiology and pathogenesis is still unknown. It is necessary to grasp the pathological changes of ALHE to distinguish it from other diseases.

Influence on the adhesion and growth of dermal papilla cells by chondroitin sulfate and heparin sulfate / 第三军医大学学报

Objective　To investigate the actions of extra cellular medium in growth and differentiation of hair follicle and to look for growth adjusting factors for dermal papilla cells (DPC). Methods　Dermal papilla cells were isolated and cultivated with two steps method and the cells were identified by immunohistochemical staining for actin. Influence was examined on the adhesion and growth of dermal papilla cells by chondroitin sulfate A, chondroitin sulfate C and heparin sulfate. Results　Two steps method of enzyme digestion for isolating and cultivating dermal papilla cells was an efficient method and large amount of dermal papilla of high purity were harvested with this method. The method is very simple and easy to manege with. Increased adhesion and growth of dermal papilla cells were observed in specimen treated with chondroitin A and heparin sulfate. No significant effects was observed in the cells treated with chondroit in sulfate C. Conclusion　Some extra cellular medium can regulate the adhesion and growth of dermal papilla cells and therefore influence the growth and development of hair follicle.

Experimental study of cytotoxic T lymphocyte associated molecule-4Ig for prevention of mice autoimmune hepatitis / 第三军医大学学报

Objective　To investigate the role of cytotoxic T lymphocyte associated molecule-4Ig(CTLA-4Ig) in the prevention of C57BL/6 mice autoimmune hepatitis. Methods　The C57BL/6 mice were intraperitoneally immunized with C57BL/6 mice liver-specific protein in complete Freund's adjuvant. At the same time CTLA-4Ig were given to observe the pathologic alteration of C57BL/6 mice liver. Results　With the increase of time of immunization, the results in the treatment group were similar to those of the control group; but inflammatory cell infiltration, hepatic cell swelling, focal necrosis and severe hepatocyte damage were found in the pathologic model group. There was a significant difference between the pathologic model group and control one. Conclusion　Autoimmune hepatitis of C57BL/6 mice can be effectively prevented by CTLA-4Ig.

Study on serum-free culture of dermal papillae cells of human hair / 中华皮肤科杂志

Objective To explore the feasibility of culturing dermal papillae cells (DPC) of hu- man hair in a serum-flee medium,and to observe the growth characteristics of these cells.Methods Cell culture flasks (plates) were pretreated with fibronectin,and DPC (2nd passage) were incubated with Williams E serum-flee medium supplemented with insulin-transferrin-selenite (ITS).Cells were observed by an inverted phase-contrast microscope.Proliferation of DPC was evaluated with 3-(4,5-dimethylthia- zol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and by their growth curve.Results In a serum-free medium,2nd passage DPC adhered to the flask surface within two to four hours of incubation; two to three days later,confluence,of the cells was observed,without noticeable proliferation.Four days later,cell connection was interrupted,isolated cells or cell clusters were seen,and detachment of some cells from the flask surface was observed.One to two weeks later,most cells had died.After incubation with 4% bovine serum for ten hours,cell proliferation was observed surrounding the remaining viable cell colonies. DPC growth curve showed stagnant phase and slow growth phase;however,log growth phase was not ob- served.Conclusion DPC could be successfully cultured in serum-free medium.However,the culture con- dition needs to be further optimized.

Cell-penetrating effects of HPV16E7_(49-57) epitope modified by peptide HIV-Tat_(49-57) / 中华皮肤科杂志

Objective To investigate the effects of the intracellular delivery of HPV16E7_(49-57) epi- tope modified by cell-penetrating peptide HIV-Tat_(49-57) and its influential factors.Methods The unique HLA-A2+/H-2kb+ limited CTL epitope of HPV16E7 fused with a cell-penetrating sequence (HIV-Tat_(49-57)) was designed and a 18-mer peptide was synthesized with aid of polypeptide solid phase synthesis technique. The intracellular transport capabilities of these peptides were tested with indirect immunofluorescence assay and laser confocal microscopy.In addition,the CTL epitope and 18-met peptide were used to stimulate PBMC of healthy C57BL/6 mice,and the E7_(49-57) specific CTL responses were measured by LDH cytotoxicity detection kit in PBMC of those mice.Results The HIV-Tat_(49-57) peptide could efficiently assist HPV 16E7_(49-57) peptide in penetrating into BHK cells in a time and dose-dependent manner (P＜0.05).Further- more,the specific CTL activity induced by the 18-mer peptide was much stronger than that induced by the single epitope.Conclusion The results show that HIV-Tat_(49-57) can efficiently deliver the exogenous anti- genic peptide into the cytoplasm of live cells,and induce specific CTL response.This is a new way to de- sign peptide-based vaccine of HPV.