Evaluation of attenuated Salmonella choleraesuis-mediated inhibin recombinant DNA vaccine in rats.

DNA vaccination has been studied intensively as a potential vaccine technology. We evaluated the effect of an attenuated Salmonella choleraesuis-mediated inhibin DNA vaccine in rats. First, 15 rats were treated with different doses of an inhibin vaccine to evaluate vaccine safety. Next, 30 rats were divided into 3 groups and injected intramuscularly with the inhibin vaccine two (T1) or three times (T2) or with control bacteria (Con) at 4-week intervals. The inhibin antibody levels increased [positive/negative well (P/N) value: T1 vs Con = 2.39 ± 0.01 vs 1.08 ± 0.1; T2 vs Con = 2.36 ± 0.1 vs 1.08 ± 0.1, P < 0.05] at week 2 and were maintained at a high level in T1 and T2 until week 8, although a small decrease in T2 was observed at week 10. Rats in the T1 group showed more corpora lutea compared with the Con group (10.50 ± 0.87 vs 7.4 ± 0.51, P < 0.05). Estradiol (0.439 ± 0.052 vs 0.719 ± 0.063 ng/mL, P < 0.05) and progesterone (1.315 ± 0.2 vs 0.737 ± 0.11 ng/mL, P < 0.05) levels differed significantly at metestrus after week 10 between rats in the T1 and Con groups. However, there were no significant differences in body, ovary, uterus weights, or pathological signs in the ovaries after immunization, indicating that this vaccine is safe. In conclusion, the attenuated S. choleraesuis-mediated inhibin vaccine may be an alternative to naked inhibin plasmids for stimulating ovarian follicular development to increase the ovulation rate in rats.

Regulation of cell growth by selective COX-2 inhibitors in oral carcinoma cell lines.

Evidence indicates that NSAIDs that inhibit prostaglandin (PG) synthesis can reduce the incidence of colorectal cancers and that inhibition of cyclooxygenase-2 (COX-2) may be the underlying mechanism. The objective of this study was to investigate this putative mechanism by examining the effect of selective COX-2 inhibitors (Celebrex, DFU, NS-398) and COX-1 inhibitors (Aspirin) on the growth of two human oral carcinoma cell lines (OEC-M1 and KB) and one normal fibroblast cell line (NF). We found that the growth of OEC-M1 cells could be significantly inhibited by DFU concentrations above 30 microM (31%) after 4 days, and above 50 microM (35%) after 2 days in culture; by Celebrex at concentrations above 20 microM (52%) after 6 days, above 30 microM (36%) after 5 days, and above 40 microM (33%) after 4 days in culture; and by NS-398 above 1 microM (30%) after 6 days, and above 10 microM (35%) after 5 days in culture. The growth of KB cells could be significantly inhibited by DFU concentrations above 10 microM (33%) after 6 days, above 20 microM (35%) after 4 days in culture; and by Celebrex at concentrations above 10 microM (33%) after 5 days, and above 50 microM (30%) after 4 days in culture; and by NS-398 above 1 microM (45%) after 5 days, above 20 microM (36%) after 4 days in culture. The growth of NF cells could be significantly inhibited by DFU above 30 microM (45%) after 6 days, and above 40 microM (32%) after 3 days in culture, and by Celebrex at concentrations above 10 microM (42%) after 6 days, above 30 microM (31%) after 4 days, above 50 microM (32%) after 3 days in culture, and by NS-398 above 0.1 microM (35%) after 4 days, and above 1 microM (32%) after 3 days in culture. The growth-inhibitory concentration (IC50) values for DFU on OEC-M1, KB, and NF cells were about 39.1, 14.8, and 42.9 microM at 144 h, respectively, and on KB was about 45.2 microM at 120 h. The IC50 values for Celebrex on OEC-M1, KB, and NF cells were about 19.1, 8.6, and 15.8 microM at 144 h, respectively, and on KB and NF were about 27.7 and 35.3 microM, respectively, at 120 h. The IC50 values for NS-398 on OEC-M1, KB, and NF were about 18.9, 0.7 and 1 microM, respectively, at 144 h; on KB and NF values were about 10.8 and 1.4 microM, respectively, at 120 h and on KB and NF were about 26.6 and 4.1 microM, respectively, at 96 h. The results show that the growth of these cell lines is inhibited by three COX-2 selective inhibitors but not by any COX-1 selective inhibitors. These findings suggest that COX-2 may play an important role in the generation of biochemical mediators that stimulate the growth of human oral cancer and normal fibroblast cell lines.

Differential expression of the suppressor PML and Ki-67 identifies three subtypes of human nasopharyngeal carcinoma.

The promyelocytic leukaemia (PML) gene, which encodes a transformation and growth suppressor, was found to regulate transcription and apoptosis. PML was first identified at the chromosomal translocation break-points t(15;17) of acute promyelocytic leukaemia and the gene product may mediate cell-cycle control and apoptosis. PML was found to interact with the co-transactivator CREB binding protein (CBP) and the apoptotic-modulator Bax. To determine if PML, CBP and Bax may be involved in solid tumours, such as the nasopharyngeal carcinoma (NPC), a rare neoplasia that is prevalent in Southern China, the expression of these proteins and the proliferation marker Ki-67 was analysed by immunohistochemical staining. Expression of PML in the PML-oncogenic domain (POD) or nuclear bodies in most NPC was inversely correlated with the expression of Ki-67. In addition, based on PML expression patterns in NPC three subtypes could be identified, namely, Subtype-1, with strong PML expression in POD structures and with low Ki-67 staining; Subtype-2, where PML was expressed in a homogeneously diffused pattern, but with a low intensity in the tumour cells; while Ki-67 was expressed in a moderate number of cells and Subtype-3, where the majority of tumour cells were PML-negative, while a considerable number of tumour cells were strongly labelled with Ki-67. Furthermore, CBP was present in most of the NPC cells with moderate-strong nuclear staining, while the expression in non-tumour cells were relatively weak. However, there was no direct correlation between PML and CBP expression in the NPC examined. In addition, there was low or no expression of Bax in the NP and NPC. This is, to our knowledge, the first report describing PML and CBP expression in NPC and our data strongly suggests that PML and CBP, but not Bax, may play a role in the transformed phenotypes of NPC.

The effect of betel nut extract on cell growth and prostaglandin endoperoxide synthase in human epidermoid carcinoma cells.

The objective of this study was to find out whether prostaglandin endoperoxide synthase (PHS) involves the action of betel nut extract (BNE) on the growth of oral cancers. Therefore, growth and PHS activity were examined in two human oral carcinoma cell lines (OEC-M1 and KB) and one normal fibroblast cell line (NF) in the presence of increasing BNE concentration. BNE at concentrations above 50 microg/ml significantly inhibited the cell growth of OEC-M1 after 72 h in culture, of KB and NF after 48 h in culture. The IC50 of BNE in OEC-M1, KB and NF at 24 h in culture was about 406, 37.5 and 140 microg/ml respectively. PHS activity in OEC-M1 was significantly increased by low BNE concentrations (50 microg/ml, 114%; 100 microg/ml, 33%; 150 microg/ml, 30%) but significantly reduced at higher BNE concentrations (300 microg/ml, 33%; 500 microg/ml, 61%). The PHS activity in KB was significantly inhibited by BNE and this effect was intensified as concentrations increased (50 microg/ml, 31%; 100 microg/ml, 24%; 150 microg/ml, 43%; 300 microg/ml, 60%; 500 microg/ml, 92%). Similar to that in OEC-M1, the PHS activity in NF was significantly increased at low BNE concentrations (50 microg/ml, 139%; 100 microg/ml, 87%;150 microg/ml, 77%) but reduced at higher concentrations (300 microg/ml, 55%; 500 microg/ml, 72%). The PHS activity in all cell lines was almost completely blocked by indomethacin (5 x 10(-6) M). We conclude that these findings suggest that PHS may be an important biochemical mediator of the effect of BNE on the growth of two human oral carcinoma cell lines.

Transcriptional activation of NF-kappa B activity by Epstein-Barr virus (EBV) LMP1 as a selective therapeutic strategy for EBV-associated diseases.

Epstein-Barr virus (EBV) has been known to be associated with many malignant tumors, including nasopharyngeal carcinoma (NPC). Previous studies have indicated that an EBV-encoded oncoprotein, latent membrane protein 1 (LMP1), is expressed in many NPC tissues. LMP1 has been shown to stimulate HIV LTR through the two NF-kappa B binding sites within this promoter. In this study, we examined the feasibility of using this property of LMP1 as a therapeutic strategy for the treatment of NPC. This therapy consists of the preferential killing of the LMP1-expressing cells by gene transfer using the NF-kappa B-mediated herpes simplex virus thymidine kinase (HSVtk)/ganciclovir (GCV) system. The 800-bp HIV-LTR, which contains two NF-kappa B binding sites, was used to drive the HSVtk gene. Stable C33A cell clones expressing the LMP1 and the HSVtk genes were subjected to the GCV sensitivity test. Results showed that cells expressing both the LMP1 and the HSVtk genes were highly sensitive to GCV treatment. These cells were introduced into nude mice subcutaneously and tumors became palpable within 2 weeks. GCV was then introduced intraperitoneally to these mice and the sizes of the tumors were measured daily. Results showed that the tumors regressed in the group of mice carrying cells that stably expressed both the LMP1 and the HSVtk genes, but not in mice carrying cells containing LMP1 or HSVtk alone. Our data indicate that the HSVtk gene expressed from a NF-kappa B-binding motif-containing promoter that is regulated by LMP1 may be used as an in vivo gene therapy strategy of EBV LMP1-expressing cancers such as NPC.

Bond strength with APF applied after acid etching.

One tenet of successful orthodontic therapy is to complete treatment without decalcification, hypocalcification, or discoloration of the natural dentition. Fluoride application has been shown to reduce demineralization of enamel. The purpose of this study was to see if fluoride could be incorporated into enamel before orthodontic bracketing without adversely affecting bond strength. Forty extract adolescent human premolars were randomly divided into two equal groups with 20 teeth each. Group 1 served as control group, and group 2 (experimental) was immersed in 1.23% acidulated phosphate fluoride for 4 minutes after acid etching. The buccal surfaces of all 40 teeth were then bonded with the same type of metal bracket and debonded with an Instron machine. The debonding interface was observed with scanning electron microscopy (SEM). The mapping was calculated with energy dispersive x-ray spectrometry. The results showed that the bond strength of group 1 was significantly greater than that of group 2. The enamel detachment (enamel fracture) was found in the experimental group only. Although the application of acidulated phosphate fluoride to a tooth can prevent dental decay or decalcification, the bond strength decreases and enamel detachment is found after debonding. The result shows that the application of acidulated phosphate fluoride after acid etching enamel has an adverse effect on orthodontic bond strength of human enamel.

Epstein-Barr virus LMP1 modulates the malignant potential of gastric carcinoma cells involving apoptosis.

About 10% of gastric carcinomas including lymphoepithelioma-like carcinoma and adenocarcinoma are associated with Epstein-Barr virus (EBV) infection. In EBV-associated gastric carcinomas, the tumor cells express Epstein-Barr nuclear antigen 1 (EBNA-1) but not EBNA-2, -3A, -3B, or -3C, leader protein, or latent membrane proteins (LMPs) because of gene methylation. Only a few exceptional cases have LMP1 expression in tumor cells as demonstrated by immunohistochemical studies. To elucidate the biological effects of LMP1 and the significance of its restricted expression in EBV-associated gastric carcinomas, the LMP1 gene was transferred into EBV-negative gastric carcinoma cell lines (SCM1 and TMC1) and into EBV-negative nasopharyngeal carcinoma (NPC) cells (HONE-1) as a control. The biological effects of LMP1 in gastric carcinoma cells were monitored in vitro and in vivo. These results showed that the consequence of LMP1 expression is a growth enhancement in NPC cells, but it is a growth suppression in gastric carcinoma cells. The LMP1-expressing gastric carcinoma cells had a reduced growth rate, colony-forming efficiency, mean colony size, and tumorigenicity and a lower malignant cytological grade. The reduced growth rate, colony-forming efficiency, and mean colony size were partially reversible in vitro with treatment with LMP1 antisense oligonucleotide. In addition, enhanced apoptosis was found in the LMP1-expressing gastric carcinoma cells. This suggests that LMP1 may negatively modulate the malignant potential of gastric carcinoma cells via an enhancement of apoptosis. We concluded that the restriction of LMP1 expression in EBV-associated gastric carcinomas may lead to a growth advantage for tumor cells by avoiding LMP1 apoptotic effects and immunologically mediated elimination.

Inhibition of the synthesis of eicosanoid-like substances in a human oral cancer cell line by interferon-gamma and eicosapentaenoic acid.

The objectives were to examine the production of eicosanoids in a Chinese human oral cancer cell line (OEC-M1) and to test the effects of interferon-gamma (IFN-gamma), eicosapentaenoic acid (EPA) and enzyme inhibitors on this biosynthesis. The eicosanoids were identified by reverse phase-high performance liquid chromatography. Two predominant peaks appeared in the chromatograms. One compound (P-1) was identified by ultraviolet absorption at a lambda(max) of 278nm with shoulders at 272 and 284nm. The other compound (P-2) was identified by ultraviolet absorption at a lambda(max) of 284 nm with shoulders at 278 and 290 nm. The production of P- was significantly inhibited by the addition of IFN-gamma (200 and 400 U/ml), and EPA (10 to 40 microM). It was only partially inhibited (p < 0.05) by indomethacin (INDO) (0.5 and 1 microM), nordihydroguaiaretic acid (NDGA) (30 and 60 microM/ml), and eicosa-5,8,11,14-tetraynoic acid (ETYA) (20-60 microM). It was almost completely inhibited by indomethacin (2 and 3 microM), and dexamethasone (0.6 and 6 microM). The production of P-2 was almost completely inhibited by IFN-gamma (200 and 400 U/ml), and partially inhibited (p < 0.05) by EPA (10 and 20 microM), NDGA (30 and 60 microM), ETYA (20 and 40 microM), dexamethasone (0.6 and 6 microM). The production of both peaks was significantly reduced by excluding arachidonic acid (AA), and almost completely inhibited by heating at 100 degrees C for 10 min during incubation. These results demonstrate that two eicosanoid-like compounds are synthesized by the OEC-M cell line and that their production can be modulated by IFN-gamma, EPA, indomethacin, NDGA, ETYA, and dexamethasone.

Sequence variations between two Epstein-Barr virus LMP 1 variants have no effect on the activation of NF-kappaB activity.

Previously, we reported that the LMP 1 gene of Epstein-Barr virus (EBV) derived from nasopharyngeal carcinoma (NPC) tissues (i.e., NLMP 1 gene) was able to transform BALB/c3T3 cells. On the other hand, LMP 1 gene of B95-8 strain (i.e., BLMP 1 gene) was not able to transform these cells (Chen et aL, 1992). Further studies indicated that a 10-amino-acid deletion in the carboxyl terminus of NLMP 1 played an important role in transformation (Li et al., 1996). In this study, we tested if this 10-amino-acid deletion affected the induction of NF-kappaB activity by LMP 1. The long terminal repeat of the human immunodeficiency virus type 1 (HIV-1 LTR) contained two copies of NF-kappaB sites and was used to construct the Luc gene-based reporter plasmid, p kappaB-Luc. Plasmid p kappaB-Luc was co-transfected with plasmids containing the NLMP 1 gene, BLMP 1 gene, and their chimeric or deletion constructs, respectively, into C-33A and BALB/c3T3 cells. The activation was then measured by the luciferase activity. Results showed that the full-length proteins induced a similar level of NF-kappaB activity, the two 3' mutants (R15delta and D4delta) still induced a relatively high level of activity, and the two 5' deletion mutants (delta3058 and delta3243) of NLMP 1 gene did not show any significant activation in C-33A cells. However, none of these LMP 1 proteins induced NF-kappaB activity in BALB/c3T3 cells. Using subcellular fractionation analysis and an immunocytostaining method, the truncated proteins of delta3058 and delta3243 were detected in the cytoplasm of the cells whereas the full-length NLMP 1 protein was located at the cytoplasmic membrane. Stable BALB/c3T3 cell clones that expressed both truncated proteins were established and then their ability to induce tumors in nude mice was examined. Data showed that both truncated NLMP 1 proteins still maintained partial transformation activity. Our results suggested that there was no direct correlation between NF-kappaB activation and transformation activity of LMP 1 in BALB/c3T3 cell transformation and that the amino-terminal membrane-spanning domain was important for maintaining both functions of LMP 1.

Fluoridated etching on orthodontic bonding.

Enamel demineralization that occurs adjacent to directly bonded orthodontic attachments is of great concern to orthodontists. One procedure suggested to overcome this problem is to use fluoride treatment in an acid etch. The purpose of this study was to evaluate the bond strength and the debonding interface distribution of adhesive, with and without the use of fluoridated etch on enamel, before bonding. Ten teeth were etched with 37% phosphoric acid (H3PO4), incorporated with 1.23% sodium fluoride (NaF) for 15 seconds. The control group of 10 teeth was etched with the 37% H3PO4 solution for 15 seconds without fluoride. Fluoride on enamel was first detected with scanning auger microscribe/photoelectron spectroscopy in the fluoridated etching group. The brackets were then bonded on the labial surfaces of the crowns of both groups of teeth. The bracketed teeth were tested, with an Instron machine, to determine the tensile bond strength, as well as with a scanning electron microscope, and by mapping with an energy dispersive x-ray spectrometer to detect the debonding interfaces. The results showed that the fluoride was found on the enamel after fluoridated etching for 15 seconds. The bond strength and debonding interface distribution between the two groups were not statistically significantly different. Enamel detachment was not present in either group. Hence, the fluoridated etching with 1.23% NaF may have a clinical application in the prevention of demineralization or caries surrounding and under orthodontic brackets bonded to enamel.

Analysis of bcl-2 expression in normal, inflamed, dysplastic nasopharyngeal epithelia, and nasopharyngeal carcinoma: association with p53 expression.

To further characterize bcl-2 expression in nasopharyngeal carcinoma (NPC), the authors analyzed bcl-2 expression immunohistochemically in biopsy specimens from 101 cases of NPC, of which 65 had the component of normal nasopharyngeal epithelium (NPE), 24 with dysplastic lesions adjacent to carcinoma, and 14 with both primary and metastatic lesions. An additional 25 nasopharyngeal biopsies of NPE from patients with chronic inflammation of nasopharynx were also included. The percentage of detectable bcl-2 expression shown in NPC (80%) and adjacent dysplastic lesions (71%) was significantly higher than in adjacent NPE (37%) and NPE from patients with chronic inflammation of the nasopharynx (30%) (P < .05). In both normal and inflamed NPE, the detectable bcl-2 expression was restricted to the basal cells; however, in dysplastic lesions, the bcl-2 staining distribution was increased with the dysplastic cell layers, and in entire layers of epithelium in severe dysplasia or carcinoma in situ. In addition, the staining intensity of bcl-2 in carcinomas and adjacent dysplastic lesions was generally stronger than that of adjacent NPE. These observations suggest that the expression of bcl-2 in dysplasia and carcinoma is enhanced relative to that of adjacent NPE. Enhanced bcl-2 expression to prevent apoptosis seems to occur from the early stages and may play an important role in the carcinogenesis of NPC. Furthermore, up to 77% of NPC with the coexpression of bcl-2 and p53 was observed and suggested that the association of bcl-2 and p53 expression seems to occur from the early stages of the development of NPC. The overexpression of p53 protein in NPC suggests that the mutation of p53 gene or altered function of wild-type p53 protein may contribute to the pathogenesis. It is conceivable that the presence of both enhanced bcl-2 expression and altered p53 functions may play a crucial synergistic effect in the carcinogenesis of NPC.

Bond strength: a comparison between chemical coated and mechanical interlock bases of ceramic and metal brackets.

Two types of chemically coated bases, two types of mechanical interlock base polycrystalline ceramic brackets, as well as one type of mechanical interlock base metal bracket were selected for bonding with Concise orthodontic resin on 60 extracted premolars. Bond strength was measured with an Instron testing machine and the debonded interface and enamel detachment were examined with scanning electron microscope and energy dispersive x-ray spectrometer. The results showed the greater bond strength with a chemically coated base of ceramic brackets had a greater debonded interface between enamel and resin, and the weaker bond strength of mechanical interlock base of ceramic and metal brackets had a greater debonded interfaces between bracket and resin. There was no significant statistical difference in bond strengths with mechanically interlock bases between ceramic and metal brackets. The enamel detachment was found on only the stronger bond strength in which there was a chemically coated base on the ceramic bracket. Ceramic bracket fractures were not found during debonding in this specially designed specimen with 1 mm/min speed of crosshead. The mechanical interlock base of the ceramic bracket combines the strength, durability and retention of a metal bracket along with an aesthetic advantage and no enamel detachment after debonding.

Enhanced malignant progression of nasopharyngeal carcinoma cells mediated by the expression of Epstein-Barr nuclear antigen 1 in vivo.

Nasopharyngeal carcinoma (NPC) is consistently associated with Epstein-Barr virus (EBV) and mostly classified as poorly differentiated squamous cell carcinoma or undifferentiated carcinoma with early metastasis and a rapidly progressive clinical course. The EBV-encoded latent proteins, Epstein-Barr nuclear antigen 1 (EBNA 1) and latent membrane proteins (LMPs), may be expressed in NPC, but their biological effects are poorly understood. EBNA 1 may predispose B lymphocytes to lymphomagenesis in transgenic mice, but its biological effects in NPC are still unknown. This study investigated the biological effects of EBNA 1 by expressing it in an EBV-negative NPC cell line (HONE-1), which was then inoculated into both nude and severe combined immunodeficiency mice. The EBNA 1 caused HONE-1 cells to grow in a less differentiated pattern and to progress more rapidly, as well as increasing their tumourigenicity and metastatic capability. These data suggest that EBNA 1 may play a critical role in the progressive evolution of NPC.

Expression of p53 protein in colorectal carcinoids.

OBJECTIVE: To determine the frequency and prognostic significance of p53 protein expression in colorectal carcinoid tumors. DESIGN: Thirty-one paraffin-embedded specimens of colorectal carcinoid tumor were studied by immunohistochemical staining to detect p53 protein expression. The association of p53 expression with tumor site, tumor size, invasion level, tumor stage, DNA pattern, and patient survival were analyzed. RESULTS: p53 protein was detected in five (16%) of 31 colorectal carcinoid tumors. There was a correlation between p53 overexpression and tumor site, tumor size, tumor stage, and DNA ploidy (P < .05) but not for the depth of tumor invasion (P = .06). In addition to tumor size, invasion, stage, and DNA aneuploidy, p53 protein overexpression was also indicative of a poor prognosis (P < .001). CONCLUSIONS: The overexpression of p53 protein is uncommon in colorectal carcinoid tumors. However, the expression of p53 protein has a correlation with clinicopathologic-predicting criteria in colorectal carcinoid tumors and may be used as an associated prognostic parameter to assess patient survival.

Optimal dose of garlic to inhibit dimethylhydrazine-induced colon cancer.

This study was designed to investigate the optimal dose of garlic during long-term feeding and its preventive and therapeutic effects on colon cancer in rats induced by 1,2-dimethylhydrazine (DMH). A total of 240 male Sprague-Dawley rats were grouped and fed with either a basal or a garlic diet of different concentration, and some groups were subcutaneously injected with DMH 20 mg/kg once a week for 20 weeks. The incidence of colon tumor was significantly decreased in the groups fed with 2.5%, 5%, and 10% garlic diets (p < 0.001). There was no distinct difference among these concentrations (p > 0.05). Therefore the minimal optimal dose of garlic to inhibit colon cancer was 2.5%. The equivalent dose of this concentration in humans is 4.76 g/m2 body surface/day. In a therapeutic study, the tumor-inducing interval in nude mice subcutaneously injected with colon cancer cells (CC-M2) was prolonged by a 2.5% garlic diet (p < 0.01). Thus smaller tumor volume and longer survival time were found in the garlic group than in the controls (p < 0.01). However, the growth rate of tumors was not markedly inhibited by garlic. All rats finally died within 18 weeks. This study suggested that a 2.5% garlic dose may be used mainly as an inhibitor to prevent colon cancers and improve survival time.

Detection of human papillomavirus DNA in colorectal carcinomas by polymerase chain reaction.

Human papillomaviruses (HPVs) are associated with a number of benign and malignant neoplasms. To substantiate the relationship between HPV DNA and colorectal carcinomas, 70 carcinomas and 37 adenomas were analysed in this study. Specific types of HPV DNA in colorectal tumours were detected by polymerase chain reaction (PCR) and Southern blot hybridisation. HPV DNA was detected in 11 of 37 (29.7%) adenomas and in 52.9% 37 of 70 (52.9%) of carcinomas. The expression of HPV DNA in adenomas and carcinomas, especially that of HPV 16 in HPV positive cases (4 of 11 v 26 of 37), was significantly different (p < 0.05). There was no correlation, however, between HPV and the location, differentiation, stage, or survival of malignant neoplasms. These data suggest that HPV DNA, especially type 16, is associated with colorectal carcinogenesis.

Assessment of p53 expression in nasopharyngeal carcinoma.

We analyzed the expression of the p53 protein by immunohistochemical methods from 101 patients with nasopharyngeal carcinoma (NPC): 24 with NPC and dysplastic lesions adjacent to carcinoma and 14 with primary and metastatic specimens. Ninety-six of 101 lesions (95%) had detectable p53 protein in the nuclei of tumor cells, indicating that overexpression of the p53 protein might be closely associated with NPC. Among 24 patients who had NPC and dysplastic lesions adjacent to carcinoma, 19 of the dysplastic lesions (79.2%) and 22 of the carcinomas (91.7%) showed positive staining for the p53 protein. In dysplastic epithelia p53 antigenicity was generally in a basal location. The significant association of p53 expression in NPC and dysplastic lesions adjacent to carcinoma (P < .0001, Fisher's exact probability test) suggests that p53 overexpression seems to occur at an early stage in the development of NPC. p53 expression in NPC does not correlate with histological grading, degree of lymphocytic infiltration between tumor cells, clinical stage, sex, or age (P > .05, chi-squared test). A comparison of p53 expression between primary and metastatic NPC was performed in 14 lesions. Although the p53 protein was consistently expressed in primary and metastatic tumor cells, there was no significant difference in p53 expression in both distinct but related lesions (P > .05, paired t-test). Our results suggest that the association of overexpression of the p53 protein in NPC may not be indicative of a mutant type p53 protein.

Detection of human papillomavirus DNA in colorectal adenomas.

OBJECTIVE: To assess the presence of different types of human papillomavirus (HPV) DNA in colorectal adenomas. DESIGN: The extracted DNA of 109 formalin-fixed, paraffin-embedded tissue sections of colorectal adenomas were analyzed by polymerase chain reaction and Southern blot hybridization. The correlations of HPV types 6, 11, 16, 18, and 33 DNA with the histological patterns of adenomas were also analyzed. RESULTS: Human papillomavirus DNA was detected in 28% of the adenomas. There were eight (21%) of 38 in tubular adenomas, 13 (33%) of 40 in tubulovillous adenomas, and 10 (32%) of 31 in villous adenomas. All HPV-6/11-positive cases were tubular or tubulovillous adenomas. However, most HPV-16 infections (8/12) were seen in villous adenomas. Human papillomavirus-positive adenomas included three (8%) of 38 that showed mild dysplasia, 10 (25%) of 40 that showed moderate dysplasia, and 18 (58%) of 31 that showed severe dysplasia. CONCLUSION: The association of the histological type with HPV-16 and the association of the grade of epithelial dysplasia with HPV DNA were highly significant. These associations support the adenoma-carcinoma hypothesis. In addition, the results suggest that HPV infection may be an important factor for the development of colorectal neoplasia.

Orthodontic resin under water immersion.

The absorption and desorption of water by a polymer matrix of composite orthodontic resin could cause debonding of the filler-matrix or hydrolytic degradation of fillers and loss of bond strength. In this study, the bond strength of brackets directly bonded with orthodontic composite to the enamel surface of premolars was measured with an Instron machine; the debonding interface distribution was analyzed by scanning electron microscope and energy dispersive x-ray spectrometry following water immersion for 1, 2, and 3 days, and 1, 2, 4, 8, 16, 24, and 32 weeks, respectively. The results show that, under water immersion, bond strength may gradually weaken over time. The greatest loss occurs initially, followed by a period of relative stabilization, and then a weaker reduction after 24 weeks. The greater the time in water immersion, the less the bond strength and the greater the destruction of the composite resin. The debonding interface occurs between bracket and resin.