Nomogram incorporating potent inflammatory indicators for overall survival estimation of patients with primary oral squamous cell carcinoma.

Background: Inflammation has been recognized to be a factor that substantially influences tumorigenesis and tumor prognosis. Hence, this study was aimed to investigate an inflammatory marker with the most potent prognostic ability and to evaluate the survival estimation capability of dynamic change in this marker for patients suffered from oral squamous cell carcinoma (OSCC). Methods: 469 patients' inflammatory indicators including lymphocyte-to-monocyte ratio (LMR), neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR) and systemic inflammatory response index (SIRI), were calculated. Their predictive abilities for overall survival (OS) were evaluated by Kaplan-Meier curves to screen for the one with the most potent prognostic value. The predictive ability of dynamic changes in this marker was verified and a predictive nomogram incorporating inflammatory indicators was developed. Results: A high LMR was identified to be an indicator of a satisfactory survival rate. Compared with that of other inflammatory markers, area under the receiver operating characteristics (ROC) curve (AUC) of LMR for 1-year and 3-year OS was significantly larger (P<0.001). Dynamic LMR change remained an significant parameter for predicting OS (OR: 2.492, 95% CI: 1.246-4.981, p = 0.010). The nomogram incorporating LMR exhibited a superior prognostic significance than the TNM system, as suggested by the C-index (0.776 vs 0.651 in primary cohort; 0.800 vs 0.707 in validation cohort, P<0.001) and AUC. Conclusions: LMR was demonstrated to possess a more potent survival estimation capability than the other three inflammatory parameters. Dynamic changes in LMR serves as a significant parameter for overall survival estimation of primary OSCC patients. The established nomogram incorporating inflammatory markers showed more accuracy and sensitivity for survival estimation of primary OSCC patients.

Activation of antitumor immune responses by Ganoderma formosanum polysaccharides in tumor-bearing mice.

Fungi of the genus Ganoderma are basidiomycetes that have been used as traditional medicine in Asia and have been shown to exhibit various pharmacological activities. We recently found that PS-F2, a polysaccharide fraction purified from the submerged culture broth of Ganoderma formosanum, stimulates the maturation of dendritic cells and primes a T helper 1 (Th1)-polarized adaptive immune response in vivo. In this study, we investigated whether the immune adjuvant function of PS-F2 can stimulate antitumor immune responses in tumor-bearing mice. Continuous intraperitoneal or oral administration of PS-F2 effectively suppressed the growth of colon 26 (C26) adenocarcinoma, B16 melanoma, and sarcoma 180 (S180) tumor cells in mice without adverse effects on the animals' health. PS-F2 did not cause direct cytotoxicity on tumor cells, and it lost the antitumor effect in mice with severe combined immunodeficiency (SCID). CD4(+) T cells, CD8(+) T cells, and serum from PS-F2-treated tumor-bearing mice all exhibited antitumor activities when adoptively transferred to naïve animals, indicating that PS-F2 treatment stimulates tumor-specific cellular and humoral immune responses. These data demonstrate that continuous administration of G. formosanum polysaccharide PS-F2 can activate host immune responses against ongoing tumor growth, suggesting that PS-F2 can potentially be developed into a preventive/therapeutic agent for cancer immunotherapy.

Polysaccharides from Ganoderma formosanum function as a Th1 adjuvant and stimulate cytotoxic T cell response in vivo.

The fungus of Ganoderma is a basidiomycete that possesses a variety of pharmacological effects and has been used in traditional Asian medicine for centuries. Ganoderma formosanum is a native Ganoderma species isolated in Taiwan, and we have previously demonstrated that PS-F2, a polysaccharide fraction purified from the submerged culture broth of G. formosanum, exhibits immunostimulatory properties in macrophages. In this study, we further characterized the adjuvant functions of PS-F2. In vitro, PS-F2 stimulated dendritic cells (DCs) to produce proinflammatory cytokines, including TNF-α, interleukin (IL)-6, and IL-12/IL-23 p40. PS-F2 also stimulated DCs to express the maturation markers CD40, CD80, CD86, and MHC class II. In a murine splenocyte culture, PS-F2 treatment resulted in elevated expression of T-bet and interferon (IFN)-γ in T lymphocytes. When used as an adjuvant in vivo with the ovalbumin (OVA) antigen, PS-F2 stimulated OVA-specific antibody production and primed IFN-γ production in OVA-specific T lymphocytes. PS-F2-adjuvated immunization also induced OVA-specific CTLs, which protected mice from a challenge with tumor cells expressing OVA. Collectively, our data show that PS-F2 functions as an adjuvant capable of inducing a Th1-polarized adaptive immune response, which would be useful in vaccines against viruses and tumors.

Extracellular polysaccharides produced by Ganoderma formosanum stimulate macrophage activation via multiple pattern-recognition receptors.

BACKGROUND: The fungus of Ganoderma is a traditional medicine in Asia with a variety of pharmacological functions including anti-cancer activities. We have purified an extracellular heteropolysaccharide fraction, PS-F2, from the submerged mycelia culture of G. formosanum and shown that PS-F2 exhibits immunostimulatory activities. In this study, we investigated the molecular mechanisms of immunostimulation by PS-F2. RESULTS: PS-F2-stimulated TNF-α production in macrophages was significantly reduced in the presence of blocking antibodies for Dectin-1 and complement receptor 3 (CR3), laminarin, or piceatannol (a spleen tyrosine kinase inhibitor), suggesting that PS-F2 recognition by macrophages is mediated by Dectin-1 and CR3 receptors. In addition, the stimulatory effect of PS-F2 was attenuated in the bone marrow-derived macrophages from C3H/HeJ mice which lack functional Toll-like receptor 4 (TLR4). PS-F2 stimulation triggered the phosphorylation of mitogen-activated protein kinases JNK, p38, and ERK, as well as the nuclear translocation of NF-κB, which all played essential roles in activating TNF-α expression. CONCLUSIONS: Our results indicate that the extracellular polysaccharides produced by G. formosanum stimulate macrophages via the engagement of multiple pattern-recognition receptors including Dectin-1, CR3 and TLR4, resulting in the activation of Syk, JNK, p38, ERK, and NK-κB and the production of TNF-α.

Polysaccharides purified from the submerged culture of Ganoderma formosanum stimulate macrophage activation and protect mice against Listeria monocytogenes infection.

The bioactive components of Ganoderma formosanum have not yet been characterized. We investigated the immunomodulatory activities of the extracellular polysaccharides produced from a submerged mycelial culture of G. formosanum. The polysaccharides were mainly composed of D-mannose, D-galactose and D-glucose. After gel filtration chromatography, three polysaccharide fractions (PS-F1, PS-F2 and PS-F3) were purified. PS-F2 stimulated mouse RAW264.7 macrophages to produce TNF-α and nitric oxide, and enhanced the phagocytic activity of macrophages. PS-F2 challenge in mice triggered an acute inflammatory response characterized by the recruitment of neutrophils and monocytes, which protected mice from subsequent infection of Listeria monocytogenes. The results indicate that the heteropolysaccharides produced by G. formosanum can activate the innate immune response on macrophages.

Formation and morphology of Zn(2)Ti(3)O(8) powders using hydrothermal process without dispersant agent or mineralizer.

Synthesis of Zn(2)Ti(3)O(8) powders for attenuating UVA using TiCl(4), Zn(NO(3))(2)·6H(2)O and NH(4)OH as precursor materials by hydrothermal process has been investigated. The X-ray diffractometry (XRD) results show the phases of ZnO, anatase TiO(2) and Zn(2)Ti(3)O(8) coexisted when the zinc titanate powders were calcined at 600 °C for 1 h. When calcined at 900 °C for 1 h, the XRD results reveal the existence of ZnO, Zn(2)TiO(4), rutile TiO(2) and ZnTiO(3). Scanning electron microscope (SEM) observations show extensive large agglomeration in the samples. Transmission electron microscope (TEM) and electron diffraction (ED) examination results indicate that ZnTiO(3) crystallites formed with a size of about 5 nm on the matrix of plate-like ZnO when calcined at 700 °C for 1 h. The calcination samples have acceptable absorbance at a wavelength of 400 nm, indicating that the zinc titanate precursor powders calcined at 700 °C for 1 h can be used as an UVA-attenuating agent.

Association of XRCC4 codon 247 polymorphism with oral cancer susceptibility in Taiwan.

BACKGROUND: The DNA repair gene XRCC4, an important caretaker of overall genome stability, is thought to play a major role in the development of human carcinogenesis. However, the association of the polymorphic variants of XRCC4 with oral cancer susceptibility has never been reported. MATERIALS AND METHODS: In this hospital-based case-control study, the association of XRCC4 codon 247 (rs3734091), G-1394T (rs6869366), intron 7 (rs28360317) and intron 7 (rs1805377) polymorphisms with oral cancer risk in a Central Taiwanese population was investigated. In total, 318 patients with oral cancer and 318 age- and gender-matched healthy controls recruited from the China Medical Hospital in Central Taiwan were genotyped. RESULTS: A significantly different distribution was found in the frequency of the XRCC4 codon 247 genotype, but not the XRCC4 G-1394T or intron 7 genotypes, between the oral cancer and control groups. A/C heterozygosity at XRCC4 codon 247 conferred a significant (2.04-fold) increased risk of oral cancer. As for XRCC4 G-1394T and intron 7 polymorphisms, there was no difference in distribution between the oral cancer and control groups. Gene-environment interactions with smoking, but not with betel quid chewing or alcohol consumption, were significant for XRCC4 codon 247 polymorphism. The XRCC4 codon 247 A/C genotype in association with smoking conferred an increased risk of 3.44 (95% confidence interval = 1.24-9.60) for oral cancer. CONCLUSION: Our results provide the first evidence that the heterozygous A allele of the XRCC4 codon 247 may be associated with the development of oral cancer and may be a novel useful marker for primary prevention and anticancer intervention.

Oral cancer and genetic polymorphism of DNA double strand break gene Ku70 in Taiwan.

The DNA repair gene Ku70, an important caretaker of the overall genome stability, is thought to play a major role in the DNA double strand break repair system. It is known that defects in double strand break repair capacity can lead to irreversible genomic instability. However, the polymorphic variants of Ku70 and their association with oral cancer susceptibility has never been reported on. In this hospital-based case-control study, the association of Ku70 promoter T-991C (rs5751129), promoter G-57C (rs2267437), promoter A-31G (rs132770), and intron3 (rs132774) polymorphisms with oral cancer risk in a Taiwanese population was investigated. In total, 318 patients with oral cancer and 318 age- and gender-matched healthy controls recruited from the China Medical Hospital in Taiwan were genotyped. The results showed that there were significant differences between the oral cancer and control groups in the distribution of their genotypes (P=0.0031) and allelic frequency (P=0.0009) in the Ku70 promoter T-991C polymorphism. Individuals who carried at least one C allele (T/C or C/C) had a 2.15-fold increased risk of developing oral cancer compared to those who carried the T/T wild-type genotype (95% CI: 1.37-3.36). In the other three polymorphisms, there was no difference between both groups in the distribution of either genotype or allelic frequency. In conclusion, the Ku70 promoter T-991C, but not the Ku70 promoter C-57G, promoter A-31G or intron3, is connected to oral cancer susceptibility. This polymorphism may be a novel useful marker for primary prevention and anticancer intervention.

A new single nucleotide polymorphism in XRCC4 gene is associated with breast cancer susceptibility in Taiwanese patients.

BACKGROUND: The DNA repair gene XRCC4, an important caretaker of the overall genome stability, is thought to play a major role in the human carcinogenesis. Some new and important polymorphic variants of XRCC4, at codon 247 (rs 3734091), G-1394T (rs 6869366), and Intron 7 (rs 28360317), and their association with breast cancer susceptibility was investigated in a Taiwanese population. MATERIALS AND METHODS: In a hospital-based case-control study, 432 female patients with breast cancer and 432 age-matched healthy controls recruited from the China Medical Hospital in Central Taiwan were genotyped. RESULTS: A significant difference in the frequency of the XRCC4 G-1394T genotype, but not the XRCC4 codon 247, or intron 7 genotypes was found between the breast cancer and control groups. Individuals with G/T or T/T at the XRCC4 G-1394T locus showed a 2.33-fold (95% confidence interval=1.37-3.98) increased risk of breast cancer compared to those with G/G. For XRCC4 codon 247 or intron 7, there was no difference in distribution between the breast cancer and control groups. CONCLUSION: Our findings suggest that the heterozygous and homozygous T allele of the XRCC4 G-1394T may be associated with the development of breast cancer and may be a useful biomarker for anticancer prevention and intervention.

A novel single nucleotide polymorphism in XRCC4 gene is associated with oral cancer susceptibility in Taiwanese patients.

The DNA double strand break repair gene XRCC4, an important caretaker of genome stability, is suggested to play a role in the development of human carcinogenesis. However, no evidence has been provided showing that XRCC4 was associated with oral oncology. In this hospital-based case-control study, the association of XRCC4 G-1394T (rs6869366), intron 3 (rs28360071), intron 7 (rs28360317), and intron 7 (rs1805377) polymorphisms with oral cancer risk in a Taiwanese population was first investigated. In total, 318 patients with oral cancer and 318 age- and gender-matched healthy controls were genotyped. We found a significant different distribution in the frequency of the XRCC4 intron 3 genotype, but not the XRCC4 G-1394T or intron 7 genotypes, between the oral cancer and control groups. Those who had heterozygous del/ins at XRCC4 intron 3 showed a 1.57-fold (95% confidence interval=1.12-2.21) increased risk of oral cancer compared to those with ins/ins. As for XRCC4 G-1394T or intron 7 polymorphisms, there was no difference in the distribution between the oral cancer and control groups. There were significant gene-environment interactions between XRCC4 intron 3 genotype with smoking and with betel quid chewing, but not with alcoholism. In smoker and betel quid chewer groups, the XRCC4 intron 3 del variants exhibited 2.57- and 3.03-fold higher risks than the ins genotype, respectively. Our results firstly suggest that the XRCC4 intron 3 del genotype may be associated with oral oncology and may be a novel useful marker for primary prevention and anticancer intervention.

A novel single nucleotide polymorphism in ERCC6 gene is associated with oral cancer susceptibility in Taiwanese patients.

The DNA repair gene ERCC6, an important caretaker of the overall genome stability, is thought to play a role in the development of human malignancy. However, the polymorphic variants of ERCC6, has never been reported about their association with oral cancer susceptibility. In this hospital-based case-control study, the association of ERCC6 codon 399, 1097 and 1413 polymorphisms with oral cancer risk in a Central Taiwanese population was first investigated. In total, 292 patients with oral cancer and 290 age- and gender-matched healthy controls recruited from the China Medical Hospital in Central Taiwan were genotyped. We found a significant different distribution in the frequency of the ERCC6 codon 399 genotypes, but not the ERCC6 codon 1097 or 1413 genotypes, between the oral cancer and control groups. Those who had homozygous A/A or heterozygous A/G at ERCC6 codon 399 showed a 1.82- and 1.22-fold (95% confidence interval=1.19-2.79 and 0.83-1.78, respectively) increased risk of oral cancer compared to those with G/G. As for ERCC6 codon 1097 or 1413, there was no difference in distribution between the oral cancer and control groups. Gene-environment interactions with smoking and betel quid chewing, but not alcohol drinking, were significant for ERCC6 polymorphisms. ERCC6 codon 399, G/A+A/A ever user groups exhibited an increased risk of 2.36 (95% CI=1.36-4.10) for smoking and 2.72 (95% CI=1.31-5.63) for betel quid chewing. Our results firstly suggest that the heterozygous and homozygous A allele of the ERCC6 codon 399 may be associated with the development of oral cancer and may be a novel useful marker for primary prevention and anticancer intervention.

Significant association of XRCC4 single nucleotide polymorphisms with prostate cancer susceptibility in Taiwanese males.

The DNA repair gene X-ray cross-complementing group 4 (XRCC4), a member of the non-homologous end-joining (NHEJ) repair system, plays a major role in the repair of the double-strand breaks of the DNA sequence. This gene is critical to the maintenance of overall genome stability, and is also thought to play a key role in human carcinogenesis. In this case-control study, several novel polymorphic variants of XRCC4, including C-1622T (rs7727691), G-1394T (rs6869366), C-571T (rs2075686) and intron3 DIP (rs28360071), were investigated, and the correlation of these variants to prostate cancer susceptibility in a Taiwanese population was observed. A total of 134 prostate cancer patients were recruited along with 134 age-matched healthy controls, and the association of their selected genotypes with susceptibility to prostate cancer was determined. The G-1394T variant of XRCC4 proved, after analysis of the frequencies of each variant in the prostate cancer and control groups, to be a significant single nucleotide polymorphism (SNP) in prostate carcinogenesis. Our data clearly indicate that the heterogeneous G of G-1394T increases the risk of suceptibility to prostate cancer (P=0.0106), while no difference in distribution of XRCC4 C-1622T (rs7727691), C-571T (rs2075686) or intron3 DIP (rs28360071) between the prostate cancer and control groups was found. In conclusion, our findings suggest that the G allele of XRCC4 G-1394T may be responsible for prostate carcinogenesis, and could be useful in the early detection and prevention of the disease.

A novel single nucleotide polymorphism in XRCC4 gene is associated with gastric cancer susceptibility in Taiwan.

BACKGROUND: The DNA repair gene XRCC4, an important caretaker of the overall genome stability, is thought to play a major role in the human carcinogenesis. We investigate some novel and important polymorphic variants of XRCC4, at codon 247 (rs3734091), G-1394T (rs6869366), intron 3 (rs28360071), and intron 7 (rs28360317), of their associated with gastric cancer susceptibility. MATERIALS AND METHODS: In this hospital-based case-control study, the association of XRCC4 polymorphisms with gastric cancer risk in a Taiwanese population was investigated. In total, 121 patients with gastric cancer and 121 age-matched healthy controls recruited were genotyped investigating these polymorphisms' association with gastric cancer susceptibility. RESULTS: We found a significant difference in the frequency of the XRCC4 G-1394T genotype, but not others, between the gastric cancer and control groups. Those who had G/T or G/G at XRCC4 G-1394T showed a 3.79-fold (95% confidence interval = 1.47-9.82) increased risk of gastric cancer compared to those with T/T. As for XRCC4 codon 247, intron 3, or intron 7, there was no difference in distribution between the gastric cancer and control groups. CONCLUSIONS: Our findings suggest that the G allele of the XRCC4 G-1394T may contribute to gastric carcinogenesis and may be useful for gastric cancer early detection and prevention.

Association of XPD polymorphisms with prostate cancer in Taiwanese patients.

BACKGROUND: The DNA repair gene XPD, an important caretaker of the overall genome stability, is thought to play a major role in the development of human malignancy. Polymorphic variants of XPD, at codon 312, 751, and other sites, have been associated with cancer susceptibility, but few studies have investigated their effect on prostate cancer risk. PATIENTS AND METHODS: In this hospital-based case-control study, the association of XPD codon 312, 751 and promoter-114 polymorphisms with prostate cancer risk in a Taiwanese population were investigated. In total, 123 patients with prostate cancer and 479 healthy controls recruited from the China Medical Hospital in Central Taiwan were genotyped. RESULTS: We found a significant difference in the frequency of the XPD codon 312 genotype, but not the XPD codon 751 or promoter-114 genotypes, between the prostate cancer and control groups. Those who had GIA or A/A at XPD codon 312 showed a 1.81-fold (95% confidence interval=1.21-2.69) increased risk of prostate cancer compared to those with GIG. As for XPD codon 312 or promoter-114, there was no difference in distribution between the prostate cancer and control groups. CONCLUSION: Our findings suggest that the heterozygous and homozygous A allele of the XPD codon 312 may be associated with the development of prostate cancer and may be a useful marker for primary prevention and anticancer intervention.

[Establishment of liver metastasis model of human primary colonic lymphoma by orthotopic transplantation].

OBJECTIVE: To provide an ideal animal model for exploring pathogenesis and experimental treatment of primary colonic lymphoma. METHODS: Primary colonic and liver metastatic lymphoma tissues were obtained from the surgical specimens,and transplanted into colonic mucosa of nude mice respectively. The tumorigenesis, invasion, metastasis and morphology of the transplanted tumor were observed. Karyotype was analyzed and DNA content was measured. RESULTS: According to the new WHO classification of malignant lymphoma, two high metastatic models (HCBL-0303 from primary lymphoma and HCBL-0304 from live metastatic lesion) of human primary colonic non-Hodgkin's B cell lymphoma in nude mice were established successfully by orthotopic transplantation. Pathological examination showed poorly differentiated non-Hodgkin's B cell lymphoma of the transplanted tumors, and immunohistochemical staining showed positive expressions of CD19, CD20 and CD22, and negative expressions of CD3 and CD7. The number of chromosome ranged from 55 to 59, and DNA index (DI) was 1.59 - 1.71 (i.e. heteroploid). In HCBL-0303,liver metastasis rate was 63.7% and lymph node metastasis rate was 56.4%. However, in HCBL-0304, both metastasis rates of liver and lymph node were 100%. The transplanted tumors grew autonomously and invasively in nude mice, and further developed hematogenous, lymphatic metastasis and intraperitoneal seeding. CONCLUSIONS: HCBL-0303 and HCBL-0304 are the first established high metastatic models of primary colonic lymphoma, and can be applied to the research on pathogenesis, invasion,metastasis and experimental therapy of human primary colonic lymphoma.

[Analysing genetic distance of national wistar rat with DNA fingerprinting].

DNA fingerprinting of Wistar rat were studied with JL-02 Mulilocus probe and Southern hybridization, and comparing with different individuals of nine groups Wistar rat from six national classical area and different groups. It was indicated that DNA fingerprinting could reflect genetic material of outbred strain rat,and were more polymorphic. The genetic distances of Wistar rat were distributed for 0.2-0.6 among different individuals within same groups, and the genetic distances of Wistar rat were distributed for 0.2-0.7 among different groups.