[Expression and clinical significance of bone morphogenetic protein 6 in human breast cancer].

OBJECTIVE: To investigate the expression pattern and clinical significance of bone morphogenetic protein 6 (BMP6) in breast tissues. METHODS: The tumor and adjacent noncancerous tissues were harvested from 36 cases of breast cancer, the expression level of BMP6 mRNA of each sample was measured by quantitative RT-PCR. Immunohistochemistry study was used to examine BMP6 protein expression in 80 cases of breast cancer, then the relationship between the expression of BMP6 and relevant clinical and pathological parameters was analyzed. RESULTS: BMP6 mRNA expression in breast cancer was significantly reduced when compared with normal breast tissues (P< 0.01), BMP6 mRNA level in estrogen receptor-positive (ER) breast cancer was distinctly higher than that in ER breast cancer. The expression of BMP6 mRNA was correlated to tumor grade (P < 0.01). The expression level of BMP6 protein in breast cancer was associated to ER and PR status, histological grade and Ki-67 status (P < 0.05), but not correlated to age, tumor size, human epidermal factor receptor 2 (Her2) status and molecular subtypes of breast cancer (P > 0.05). CONCLUSION: The ectopic expression of BMP6 may play an important role in the development and progression of breast cancer.

Anti-inflammation treatment for protection of hepatocytes and amelioration of hepatic fibrosis in rats.

Chronic inflammation is considered as an important pathophysiologic mechanism of hepatic cirrhosis, which induces hepatocyte injury and activates hepatic stellate cells (HSCs), thus resulting in hepatic fibrosis. Previous studies have reported that cyclooxygenase-2 (COX-2) inhibitor can effectively treat liver fibrosis, while somatostatin (SST) analogues inhibit the activation of HSCs. The present study aimed to investigate the effects of a COX-2 inhibitor, celecoxib, combined with a SST analogue, octreotide, for protection of hepatocytes and prevention of fibrosis in a rat model of hepatic fibrosis. Therefore, a hepatic fibrosis rat model was established following peritoneal injection of thioacetamide (TAA), and the rats were then treated with a combination of celecoxib and octreotide (TAA + C). Immunohistochemistry and western blotting assays were used to assess the expression levels of proteins associated with inflammation, epithelial-mesenchymal transition (EMT), proliferation, apoptosis and autophagy. H&E staining, transmission electron microscopy and scanning electron microscopy were used to evaluate the destruction of hepatocytes. Masson's Trichrome and Sirius Red were used to measure the degree of liver fibrosis. The results demonstrated that, compared with those of the control group, the degree of liver fibrosis and the expression of the intrahepatic inflammation factors were aggravated in the TAA group. Furthermore, the apoptosis rate, EMT and autophagy of hepatocytes were also increased in the TAA group. However, treatment with TAA + C restored the aforementioned increased levels compared with the TAA group. In conclusion, treatment of rats with the combination of celecoxib and octreotide could attenuate the progress of hepatic fibrosis via protection of hepatocytes by reducing apoptosis, EMT and autophagy in hepatocytes.

[Relationship between photosynthetic characteristics and environmental factors in leaves of Pueraria lobata].

OBJECTIVE: To study the relationship between photosynthetic characteristics and environmental factors in leaves of P. lobata. METHOD: Photosynthetic characteristics and environmental factors were measured by using CIRAS-2 portable photosynthesis system. RESULT: The apparent quantum yield in leaves was 0.0173 micromol CO2 x micromol(-1) photon. The dark respiration rate was 2.9333 micromol x m(-2) x s(-1). The light compensation point of photosynthesis was 180 micromol x m(-2) x s(-1). The light saturation point was 1600 micromol x m(-2) x s(-1). The carboxylation efficiency was 0.0338 micromol x m(-2) x s(-1). The light respiration rate was 2.5 micromol x m(-2) x s(-1). The CO2 compensation point was 100 micromol x mol(-1), The CO2 saturation point was 1 600 micromol x mol(-1). CONCLUSION: Photo flux density and air temperature are major environmental factors influencing diumal changes of net photosynthetic rate.

Modification of the method to establish a hepatic VX2 carcinoma model in rabbits.

The hepatic VX2 carcinoma model in rabbits is widely used for the preclinical study of hepatocellular carcinoma. In the present study, a modification was made to the conventional method to establish the animal model, as the conventional method gives rise to frequent tumor seeding due to the drop-out of tumor fragments. In order to evaluate each distinct method of establishing the model, the rabbits were divided into two groups: Group A (the conventional method; n=20) and group B (the modified method; n=20). All surgical details were recorded for reference. At 14 days post-surgery, contrast-enhanced computed tomography (CECT) and autopsy were conducted. Microscopic morphology of tumor cells was observed using hematoxylin and eosin (H&E) and transmission electron microscopy (TEM). Vascular endothelial growth factor (VEGF) and cluster of differentiation (CD)31 were detected via immunochemistry and reverse transcription-polymerase chain reaction. In total, 19 rabbits in each group succeeded in model establishment. Throughout the surgery, group A experienced a longer surgery time compared with group B (group A vs. group B, 22.57±1.34 vs. 20.17±1.50 min; P<0.001), an increased tumor fragment drop-out frequency (group A vs. group B, 1.84±0.96 vs. 1.16±0.38; P=0.008) and an increased peritoneal nodule incidence (group A vs. group B, 35 vs. 5%, P=0.042). As for CECT, H&E and TEM, hepatic VX2 allografts in the two groups demonstrated similar imaging presentations and tumor cell morphology. In addition, VEGF and CD31 levels did not differ between the two groups. In conclusion, the modified method for the establishment of hepatic VX2 carcinoma model in rabbits may decrease tumor fragment drop-out frequency during surgery and incidence of tumor seeding without affecting the properties of VX2 carcinoma.

Morphological and molecular variations induce mitochondrial dysfunction as a possible underlying mechanism of athletic amenorrhea.

Female athletes may experience difficulties in achieving pregnancy due to athletic amenorrhea (AA); however, the underlying mechanisms of AA remain unknown. The present study focuses on the mitochondrial alteration and its function in detecting the possible mechanism of AA. An AA rat model was established by excessive swimming. Hematoxylin and eosin staining, and transmission electron microscopic methods were performed to evaluate the morphological changes of the ovary, immunohistochemical examinations and radioimmunoassays were used to detect the reproductive hormones and corresponding receptors. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was used to test the mtDNA copy number. PCR and western blot analysis were used to test the expression of ND2. The change of morphological features of the rat ovaries revealed evident abnormalities. Particularly, the features of the mitochondria were markedly altered. In addition, reproductive hormones in the serum and tissues of AA rats were also detected to evaluate the function of the ovaries, and the levels of these hormones were significantly decreased. Furthermore, the mitochondrial DNA copy number (mtDNA) and expression of NADH dehydrogenase subunit 2 (ND2) were quantitated by qPCR or western blot analysis. Accordingly, the mtDNA copy number and expression of ND2 expression were markedly reduced in the AA rats. In conclusion, mitochondrial dysfunction in AA may affect the cellular energy supply and, therefore, result in dysfunction of the ovary. Thus, mitochondrial dysfunction may be considered as a possible underlying mechanism for the occurrence of AA.

[Separation and determination of puerarin, daidzin and daidzein in stems and leaves of Pueraria thomsonii by RP-HPLC].

OBJECTIVE: To separation and determine the contents of puerarin, daidzin and daidzein in the stems and the leaves of Pueraria thomsonii, and to provide scientific basis for developing and using of the stems and the leaves. METHOD: A RP-HPLC method was applied with a Diamonsil C18 column (4.6 mm x 150 mm, 5 microm) by gradient elution using methanol-1% glacial acetic acid solution as the mobil phase. The flow rate was 1 mL x min(-1) and the detective wavelength was 250 nm, the column temperature was 25 degrees C. RESULT: All of the three compounds showed good linearities (r >0.9995) and the recoveries were in the range of 99.0% - 101.6%. The contents of puerarin, daidzin and daidzein in the stems are higher than those in the leaves. CONCLUSION: The method was accurate and could be used to contral the quality of the stems and leaves of P. thomsonii.

Wnt/beta-catenin signaling pathway is active in pancreatic development of rat embryo.

AIM: To elucidate the role of Wnt/beta-catenin signaling pathway in pancreatic development of rat embryo. METHODS: The mRNAs of beta-catenin, APC, cyclin D1 genes were amplified by means of semiquantitative reverse transcription polymerase chain reaction (RT-PCR) from embryonic pancreas in different periods and normal pancreas of rat, respectively. Protein expression of these genes in embryonic pancreas of E14.5-E18.5 was examined by immunohistochemical method. RESULTS: In embryonic pancreas of E14.5, the transcript amplification of beta-catenin and cyclinD1 genes was detected. In embryonic pancreas of E18.5, the transcription levels of beta-catenin and cyclinD1 genes became much higher than in other periods. But in adult rat pancreas the transcription of cyclinD1 gene could not be observed. Only until E18.5, the transcript amplification of mRNA of APC gene could be detected. Surprisingly, the transcription level of APC gene became much higher in adult rat pancreas than in embryonic pancreas. By means of immunohistochemical staining, identical results were obtained to the above by RP-PCR, except for beta-catenin protein in adult rat pancreas. CONCLUSION: Active Wnt/beta-catenin signaling occurs in rat embryonic pancreas and is probably important for pancreatic development and organ formation.

Comparative proteome analysis of breast cancer and adjacent normal breast tissues in human.

Two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and matrix-assisted laser desorption/ionization tandem time-of-flight mass spectrometry (MALDI-TOF/TOF-MS), incorporated with online database searching, were performed to investigate differential proteins of breast cancer and adjacent normal breast tissues. Considering that serum albumin is abundantly presented in normal control samples, 15 differential spots detected in 11 out of 12 (91.7%) breast cancer samples were identified by online SIENA-2DPAGE database searching and MALDI-TOF/TOF-MS analysis. The results indicate that pathological changes of breast cancer are concerned with augmentation of substance metabolism, promotion of proteolytic activity, decline of activity of some inhibitors of enzymes, and so on. Some important proteins involved in the pathological process of breast cancer with changed expression may be useful biomarkers, such as alpha-1-antitrypsin, EF-1-beta, cathepsin D, TCTP, SMT3A, RPS12, and PSMA1, among which SMT3A, RPS12, and PSMA1 were first reported for breast cancer in this study.

Role of beta-catenin in hepatocarcinogenesis of rats.

BACKGROUND: Beta-catenin has two distinct roles in E-cadherin mediated cell adhesion and carcinogenesis by activating the wnt/beta-catenin signaling pathway. One occurs at the cell-adhesion site, where cadherins are linked to the actin-based cytoskeleton. The other takes place in the cytoplasm and nuclei and is thought to regulate cell transformation. We studied the role of beta-catenin in hepatocarcinogenesis of rats. METHODS: Fresh liver specimens were obtained from normal rats, and atypical hyperplasia livers and hepatocarcinoma tissues from model rats. The changes of beta-catenin in gene expression levels were detected by reverse transcriptase polymerase chain reaction (RT-PCR) in the different specimens separately. At the same time, their localization was observed immunohistochemically. RESULTS: In the normal liver specimens, beta-catenin staining was seen in the cell membrane. In liver specimens of atypical hyperplasia, beta-catenin staining occurred in the cell cytoplasm of some cells as well as in the cell membrane of others. Immunohistochemically cancerous tissues showed the presence of beta-catenin in the cytoplasm and nuclei. RT-PCR revealed that the gene expression levels of beta-catenin were same in all samples. CONCLUSIONS: The accumulation of beta-catenin in the cytoplasm and/or nuclei frequently occurs in hepatocarcinogenesis of rats. It may be an early event in the development of hepatocarcinoma of rats.

Islet formation and regeneration.

OBJECTIVE: To explore the mechanisms of differentiation and development of pancreatic endocrine cells as well as pancreatic regeneration. METHODS: Human embryonic pancreatic tissue at 7-14 weeks of gestation was collected. Diabetes mellitus rat model was induced with 65 mg/kg of streptozotocin. Insulin, glucagon, somatostatin, nestin, and cytokeratin 19 (CK19) of pancreatic tissues were observed by immunohistochemistry. RESULTS: At 9 weeks of gestation, pancreatic epithelial cells began to co-express insulin, glucagon, somatostatin, and CK19 before migration. Islet cells gradually congregated along with the increase of aging, and at 14 weeks of gestation histological examination showed islet formation. At 12 weeks of gestation, nestin-positive cells could be seen in the pancreatic mesenchyme. During early embryogenesis, islet cells of pancreatic ducts co-expressed insulin, glucagon, and somatostatin. During pancreatic regeneration after damage, nestin expression of islet cells increased. CONCLUSION: In the early stage of embryogenesis, islet cells of primary pancreatic ducts can be differentiated to multipotential endocrine cells before migration. During tissue regeneration, pancreatic stem cells may differentiate and proliferate to form pancreatic islet.

[Analysis of differential expression genes related to different metastasis potential of adenoid cystic carcinoma using restriction fragments differential display PCR].

OBJECTIVE: To construct differential expression profiles of adenoid cystic carcinoma cell lines for screening candidate genes related to metastasis and to verify some candidate genes in adenoid cystic carcinoma. METHODS: Restriction fragments differential display PCR (RFDD-PCR) was used to set up gene expression profiles of adenoid cystic carcinoma cell lines-ACC-M and ACC-2, with high and low metastasis potential respectively. Candidate genes were screened through bioinformatics analysis. Then, a gene family of these candidate genes was checked using semi-quantitative reverse transcription-PCR(RT-PCR). RESULTS: Two gene expression profiles including 5420 gene fragments were constructed, 12 genes of a family called matrix metalloproteinase genes (MMPs) were observed obvious differentially expressed between two cell lines. Results of semi-quantitative RT-PCR also identified this different expression of MMP2,MMP7,MMP9,MMP14,MMP15 and MMP24. CONCLUSION: The construction of gene expression profiles of ACC-M and ACC-2 cell lines makes the foundation for seeking the target genes of adenoid cystic carcinoma. MMP2,MMP7,MMP9 and MMP15 may be relevant with carcinogenesis, development and metastasis of adenoid cystic carcinoma, and different metastasis potential may result from different subtype of MMPs gene family.

[The role of beta-catenin in rat embryonic development and tumorigenesis].

OBJECTIVE: To investigate the role of beta-catenin as a key element of Wnt/beta-catenin signaling pathway in the development of embryonic liver and liver tumorigenesis. METHODS: Immunohistochemical method was used to examined beta-catenin proteins in 12-day (E12), 16-day (E16) embryonic rat liver, neonate rat liver, and in adult rat liver as well as in rat hepatoma. Beta-catenin mRNA was amplified in the above-mentioned samples by means of semiquantitative RT-PCR. RESULTS: Beta-catenin proteins were found in cytoplasm of E12 and E16 embryonic liver as well as in hepatoma. The expression level of beta-catenin protein in E12 embryonic liver was higher than that in E16. There were much more positive cells in E12 embryonic liver than in E16. However, no positive cell was observed expressing beta-catenin proteins in neonate rat liver and adult rat liver. The quantity of beta-catenin mRNA was the same in all samples. CONCLUSION: Wnt/beta-catenin signaling pathway was open during the development of embryonic liver and hepatic tumorigenesis. The accumulation of beta-catenin proteins in cytoplasm of embryonic liver and rat hepatic carcinoma cells might be caused not by the elevation of transcription of beta-catenin gene but by the avoidance of degeneration of beta-catenin protein.

[Isolation and purification of BMScs of GFP transgenic mouse using the method of adhering to cuture plastic in different time].

OBJECTIVE: To adopt the method of adhering to culture plastic in different time for cultivating and purifying BMSCs of green fluorescent protein (GFP) transgenic mice. METHODS: Bone marrow cells isolated from GFP transgenic mice are directly planted in culture flask and an exchange of the total volume medium is made at different time. Then the cells adhering to culture plastic are differently counted according to the cell types and are examined by immunohistochemistry using the antibodies of CD44, CD45 and CD54 in three days. Moreover, the cells after the exchange of the total volume medium in 4 hours, 8 hours and 24 hours are selected and successively subcultured down to the fifth passage. Then the result of amplification is calculated and the cells are examined by immunohistochemistry using the antibodies of CD44, CD45 and CD54. RESULTS: With the extending of the time for the first exchange of medium, the density of cells adhering to culture plastic increased accordingly, but the BMSCs proportion decreased. The cells after first exchange of medium in 4 hours had high BMSCs proportion but low BMSCs density, and the cells in 24 hours had high BMSCs density and low BMSCs proportion. However, the cells in 8-10 hours had high BMSCs density and also high BMSCs proportion. The subcultured BMSCs could stably express GFP. CONCLUSION: The method of adhering to culture plastic in different time for cultivating and purifying BMSCs of GFP transgenic mice is effective. It is suitable to make the first exchange of total volume medium in 8-10 hours. The subcultured cell has the capacity for amplification and will probably be a seed cell for the research of tissue engineering and gene therapy.

RNA sequencing reveals retinal transcriptome changes in STZ-induced diabetic rats.

The present study aimed to investigate changes in retinal gene expression in streptozotocin (STZ)­induced diabetic rats using next­generation sequencing, utilize transcriptome signatures to investigate the molecular mechanisms of diabetic retinopathy (DR), and identify novel strategies for the treatment of DR. Diabetes was chemically induced in 10­week­old male Sprague­Dawley rats using STZ. Flash­electroretinography (F­ERG) was performed to evaluate the visual function of the rats. The retinas of the rats were removed to perform high throughput RNA sequence (RNA­seq) analysis. The a­wave, b­wave, oscillatory potential 1 (OP1), OP2 and ∑OP amplitudes were significantly reduced in the diabetic group, compared with those of the control group (P<0.05). Furthermore, the implicit b­wave duration 16 weeks post­STZ induction were significantly longer in the diabetic rats, compared with the control rats (P<0.001). A total of 868 genes were identified, of which 565 were upregulated and 303 were downregulated. Among the differentially expressed genes (DEGs), 94 apoptotic genes and apoptosis regulatory genes, and 19 inflammatory genes were detected. The results of the KEGG pathway significant enrichment analysis revealed enrichment in cell adhesion molecules, complement and coagulation cascades, and antigen processing and presentation. Diabetes alters several transcripts in the retina, and RNA­seq provides novel insights into the molecular mechanisms underlying DR.

Anti-cancer effects of curcumin on lung cancer through the inhibition of EZH2 and NOTCH1.

Curcumin is potentially therapeutic for malignant diseases. The mechanisms of this effect might involve a combination of antioxidant, immunomodulatory, proapoptotic, and antiangiogenic activities. However, the exact mechanisms are not fully understood. In the present study, we provided evidences that curcumin suppressed the expression of enhancer of zeste homolog 2 (EZH2) in lung cancer cells both transcriptionally and post-transcriptionally. Curcumin inhibited the expression of EZH2 through microRNA (miR)-let 7c and miR-101. Curcumin decreased the expression of NOTCH1 through the inhibition of EZH2. There was a reciprocal regulation between EZH2 and NOTCH1 in lung cancer cells. These observations suggest that curcumin inhibits lung cancer growth and metastasis at least partly through the inhibition of EZH2 and NOTCH1.

[Preliminary analysis of retinal gene expression profile of diabetic rat].

OBJECTIVE: Establishing the retinal gene expression profiles of non-diabetic rat and diabetic rat and comparing the profiles in order to analyze the possible genes related with diabetic retinopathy. METHODS: The whole retinal transcriptional fragments of non-diabetic rat and 8-week diabetic rat were obtained by restriction fragments differential display-PCR (RFDD-PCR). Bioinformatic analysis of retinal gene expression was performed using soft wares, including Fragment Analysis. After comparison of the expression profiles, the related gene fragments of diabetic retinopathy were initially selected as the target gene of further approach. RESULTS: A total of 3639 significant fragments were obtained. By means of more than 3-fold contrast of fluorescent intensity as the differential expression standard, the authors got 840 differential fragments, accounting for 23.08% of the expressed numbers and including 5 visual related genes, 13 excitatory neruotransmitter genes and 3 inhibitory neurotransmitter genes. At the 8th week, the expression of Rhodopsin kinase, beta-arrestin, Phosducinìrod photoreceptor cGMP-gated channel and Rpe65 as well as iGlu R1-4 were down-regulated. mGluRs and GABA-Rs were all up-regulated, whereas the expression of GlyR was unchanged. CONCLUSION: These results prompt again that the changes in retinal nervous layer of rat have occurred at an early stage of diabetes. The genes expression pattern of visual related genes and excitatory and inhibitory neurotransmitters in rat diabetic retina have been involved in neuro-dysfunctions of diabetic retina.

Application of restriction fragment differential display-polymerase chain reaction in study on differential expression profiles of human diseases.

OBJECTIVE: To establish the restriction fragment differential display-polymerase chain reaction (RFDD-PCR) as an efficient technique for constructing and studying the gene expression profile of human tissues. METHODS: The tissues of mamma adenocarcinoma (T), cancerometastasis lymph node (L) and normal mammary (N) from one mammary infiltrating ductal carcinoma case were collected, and the gene expression profile of each kind of tissue was constructed using RFDD-PCR technique at equal pace according to the operating manual of Qbio-gene Company. Then all fragments of the three gene expression profiles were separated and displayed by electrophoresis. With the use of gene database at the website http://www.Qbio-gene.com/display, the authors identified the names of the probable fragments by bioinformatics analysis. Through comparison of the three profiles, the numbers and types of most differentially expressed gene fragments were displayed. RESULTS: The expression profiles of the three kinds of tissue have been constructed covering 1716 fragments of mammary adenocarcinoma, 1769 of cancerometastasis lymph nodes and 1922 of normal mammary tissue. Among these 5407 fragments, 39.39% were exactly the same. While 33.9% sequences of T and L showed differences in abundance or presence, 40.9% of T and N and 39.6% fragments of L and N were observed differentially expressed. These differentially expressed gene fragments were found to relate with metastasis, differentiation, inflammation and so on. CONCLUSION: RFDD-PCR is an efficient technique for research in human diseases genomics as a mass screening for complete gene expression profile with high-flux. Through comparison among three or more profiles, the screening for candidate genes of a certain disease can be accomplished, and there is probably a chance to identify novel gene or expressed sequence tag.

[Changes in the expression of vital cytokines for pancreatic development in early embryo].

OBJECTIVE: To clarify the changes in the expression of vital cytokines for pancreatic development in early embryo and hence provide preclinical data regarding embryonic pancreas transplanted for treatment, of diabetes mellitus. METHODS: Sample collection was conducted in accordance to the principle of informed consent. Histochemical S-ABC method was adopted in studying pancreas at 7-12 weeks of gestation, and analysis was made on vital cytokines expression as well as the differentiation and forming of pancreatic islets. RESULTS: It was found that the expression of Insulin, Glucagon, Somatostatin and Cytokeratin begins at 7 weeks, and the change is consistent with the differentiation and formation of pancreatic islets. IGF-I and F-VIII appear at 12 weeks. CONCLUSION: Insulin, Glucagon, Somatostatin, Cytokeratin, IGF-I and F-VIII play a regulatory role in the development of embryonic pancreas and are related to the differentiation and formation of endocrine and exocrine glands of pancreas. The pancreas after 12 weeks of gestation can be used as a donated graft for pancreas transplantation.

[Expression of HNF4alpha and HNF6 mRNA during the process of mouse liver development].

OBJECTIVE: To investigate the function of HNF4a and HNF6 during liver development. METHODS: The expression levels of HNF4alpha and HNF6 at E8, 9, 13, 15, 17, P1 and in adult mouse liver were detected by RT-PCR and in situ hybridization. RESULTS: RT-PCR results showed that HNF4alpha first expressed at E9, the time of liver bud formation, and lasted through all gestation and existed in adult liver. In situ hybridization showed that the expression of HNF4alpha was detected at the cells of liver cords during various stages of mouse liver development, and there were still a few HNF4alpha positive hepatocytes in adult liver. The cells of bile duct plate and biliary epithelial cells, endothelial cells, hematopoietic cells of liver were negative for HNF4alpha. The expression of HNF6 mRNA was detected in the liver at E9, the time of liver formation onset. Then, HNF6 mRNA disappeared transiently at E13, but it appeared again at E15. Its expression lasted until adult. In situ hybridization studies showed that most liver cord cells were positive for HNF6 at E9 and E15. At E17 and P1, the expression levels of liver cord cells declined, and HNF6 strongly expressed in the cells of bile duct plate and biliary epithelial cells. CONCLUSION: HNF4alpha could modulate the formation of liver bud, trigger the differentiation of hepatic stem cell towards hepatocytes, and keep the shape of hepatocytes. HNF6 might play a role at the onset of liver development, in the differentiation of hepatic stem cell towards biliary epithelial cells, and in maintaining the morphological characteristic of biliary epithelial cells.

The therapeutic impact of HNP-1 in condyloma acuminatum.

BACKGROUND: Condyloma acuminatum is one of the most commonly occurring sexually transmitted diseases. HNP1 is a small antimicrobial peptide that has been reported to have antiviral activities. AIM: Using the condyloma acuminatum tissue culture to resemble the situation more closely in vivo, we investigate the therapeutic effect of a recombinant plasmid encoding HNP1 gene in condyloma acuminatum tissue. METHODS: Recombinant plasmid DNA carrying HNP1 cDNA was constructed and identified. Then the recombinant plasmid was transfected into a condyloma acuminatum tissue fragment, and the HNP1 expression was determined on these tissue fragments by immunohistochemistry. TUNEL staining and flow cytometry techniques were used to examine cell apoptosis of condyloma acuminatum tissue. Relative real-time polymerase chain reaction was used to validate antihuman papillomavirus therapeutics of the treatment groups. RESULTS: Transfected HNP1 gene was expressed mainly in the cytoplasmic granules of the condyloma acuminatum tissues. Positive apoptotic cells were observed in condyloma acuminatum tissues transfected with the HNP1 gene. In addition, the HPV expression was lower in the HNP1 treatment tissues as compared to their corresponding control tissues. CONCLUSION: The results indicate that HNP1 can directly promote condyloma acuminatum cell apoptosis and play an antivirus role in the condyloma acuminatum tissue by limiting viral replication. These observations suggest a possible application for human HNP1 on condyloma acuminatum therapy.