Nutritional index in relation to prognosis of endometrial cancer.

Objective: Evaluate the prognostic value of the prognostic nutritional index (PNI) in patients with endometrial cancer (EC). Method: Laboratory and clinicopathological data from 370 patients who were diagnosed with EC between January 2010 and December 2021 were reviewed. The PNI was analyzed for correlations with recurrence and survival. The receiver operating characteristic curves were generated for the PNI. Optimal cut-off values were determined as the points at which the Youden index (sensitivity + specificity - 1) was maximal. Based on the results of the ROC curve analysis, the patients were grouped into high and low PNI groups. Differences in the clinicopathological characteristics between patients with high and low PNI were compared between the two groups. The effects of the prognostic factors were analyzed using univariate and multivariate Cox proportional hazards model. Results: The optimal cutoff value of the PNI was 52.74 for DFS (area under the curve: 0.817; 95% CI: 0.738-0.858, p <0.001). Significantly more patients in the low PNI group experienced recurrence (30.6% vs. 5.2%, p <0.001) and cancer-related death (17.8% vs. 2.8%, p <0.001). In multivariate analysis, PNI were independent prognostic factors for both DFS and overall survival OS. Conclusion: Low PNI was significantly associated with worse clinical outcomes in patients with EC. Our findings demonstrate that the PNI may be clinically reliable and useful as a prognostic marker for patients with EC. Further large-scale prospective studies are needed to confirm our findings.

Carboxylated Poly-L-lysine Potentially Reduces Human Sperm DNA Fragmentation after Freeze-Thawing, and Its Function Is Enhanced by Low-Dose Resveratrol.

Sperm DNA fragmentation (SDF) that occurs during the freezing-thawing of sperm may negatively impact the treatment outcomes of assisted reproductive technologies (ART). In a previous study, we developed a human sperm cryopreservation reagent containing carboxylated poly-L-lysine (CPLL) that reduced SDF after freeze-thawing compared with clinically popular cryopreservation reagents containing human serum albumin. However, it is unclear whether CPLL reduces SDF, as it differed from the constituents of the commercial cryopreservation reagents used for comparison. Therefore, here, we examined whether CPLL reduces the SDF of human sperm and evaluated reactive oxygen species (ROS) levels and lipid peroxidation (LPO), which are the causes of SDF; mitochondrial injury, ROS production; and impaired sperm motility. Furthermore, optimal antioxidants and their concentrations that could further enhance the reduction in SDF were determined for future clinical application in ART and underwent the same functional evaluations. CPLL can reduce SDF via inhibition of intracytoplasmic ROS and LPO. Furthermore, the addition of 0.1 mM resveratrol avoided the enhancement of SDF, which potentially affects mitochondrial and cytoplasmic ROS and LPO. This novel human sperm cryopreservation reagent containing CPLL and resveratrol has the potential to improve treatment outcomes in ART using frozen sperm.

Improved fertility of frozen-thawed porcine spermatozoa with 3,3-dimethylglutaric anhydride poly-L-lysine as a novel cryoprotectant.

In this study, we determined the efficacy of 3,3-dimethylglutaric anhydride poly-L-lysine (DMGA-PLL) as a cryoprotectant for porcine spermatozoa. Porcine spermatozoa were cryopreserved in a freezing extender containing 3% (v/v) glycerol and various concentrations of DMGA-PLL. At 12 h after thawing, the motility index of spermatozoa cryopreserved with 0.25% (v/v) DMGA-PLL (25.9) was significantly (P < 0.01) higher than that of spermatozoa cryopreserved with 0%, 0.125%, or 0.5% DMGA-PLL (10.0-16.3). In addition, the blastocyst formation rate of embryos derived from spermatozoa cryopreserved with 0.25% DMGA-PLL (22.8%) was significantly (P < 0.01) higher than that of embryos derived from spermatozoa cryopreserved with 0%, 0.125%, or 0.5% DMGA-PLL (7.9%-10.9%). The mean number of total piglets born to sows inseminated with spermatozoa cryopreserved without DMGA-PLL (9.0) was significantly (P < 0.05) lower than that of total piglets born to sows inseminated with spermatozoa stored at 17°C (13.8). However, when spermatozoa cryopreserved with 0.25% DMGA-PLL were used for artificial insemination, the mean number of total piglets (11.7) was not significantly different from that obtained following artificial insemination using spermatozoa stored at 17°C. The results showed the usefulness of DMGA-PLL as a cryoprotectant in the cryopreservation of porcine spermatozoa.

Cryopreservation of pig spermatozoa using carboxylated poly-L-lysine as cryoprotectant.

In this study, we cryopreserved pig spermatozoa using carboxylated poly-L-lysine (CPLL) as the cryoprotectant to determine its efficacy. Pig spermatozoa were placed in a freezing extender containing 3% (v/v) glycerol and different CPLL concentrations. The motility indices of the spermatozoa cryopreserved with 0.25% (v/v) CPLL at 6 (59.3), 9 (53.7), and 12 (26.2) h after thawing were significantly higher (P < 0.01 or P < 0.05) than those of the spermatozoa cryopreserved without CPLL (53.7, 40.1, and 17.5 at 6, 9, and 12 h after thawing, respectively). The concentration of CPLL in the freezing extender did not affect the ability of frozen-thawed spermatozoa to fertilize oocytes in vitro. However, the blastocyst formation rate of embryos derived from spermatozoa cryopreserved with 0.25% CPLL (24.6%) was significantly higher (P < 0.01) than that of embryos derived from spermatozoa cryopreserved without CPLL (11.2%). The conception rate of the sows inseminated with spermatozoa cryopreserved with 0.25% CPLL (72.2%) was not significantly different from that of the sows inseminated with spermatozoa stored at 17°C (81.3%). However, the mean number of total piglets born to the former (10.0) was significantly lower (P < 0.05) than that of total piglets born to the latter (13.4). The results showed that CPLL in the freezing extender maintained the motility of frozen-thawed pig spermatozoa and improved the in vitro development of embryos produced by in vitro fertilization. In addition, we have demonstrated that piglets could be obtained with artificial insemination using spermatozoa cryopreserved with CPLL.

Carboxylated Poly-l-Lysine as a Macromolecular Cryoprotective Agent Enables the Development of Defined and Xeno-Free Human Sperm Cryopreservation Reagents.

In human sperm cryopreservation, test yolk buffer and human serum albumin have been used as permeating macromolecular-weight cryoprotectants. In clinical reproductive medicine, human serum albumin is frequently used because of low risks of zoonoses and allergic reactions. However, the risk of allogeneic infectious diseases exists, and the supply may be unstable because human serum albumin is derived from human blood. Therefore, the development of xeno-free human sperm cryopreservative reagents that could overcome the aforementioned problems is warranted. We succeeded in developing a new xeno-free and defined sperm cryopreservation reagent containing glycerol, carboxylated poly-l-lysine, and raffinose. The cryopreservation reagent was not significantly different in terms of sperm motility, viability, and DNA fragmentation and was comparable in performance to a commercial cryopreservation reagent containing human serum albumin. Moreover, the addition of saccharides was essential for its long-term storage. These results may help elucidate the unknown function of macromolecular-weight permeating cryoprotective agents.

A liquid culture cancer spheroid model reveals low PI3K/Akt pathway activity and low adhesiveness to the extracellular matrix.

Three-dimensional (3D) cultures of cancer cells in liquid without extracellular matrix (ECM) offer in vitro models for metastasising conditions such as those in vessels and effusion. However, liquid culturing is often hindered by cell adhesiveness, which causes large cell clumps. We previously described a liquid culture material, LA717, which prevents nonclonal cell adhesion and subsequent clumping, thus allowing clonal growth of spheroids in an anchorage-independent manner. Here, we examined such liquid culture cancer spheroids for the acquisition of apical-basal polarity, sensitivity to an Akt inhibitor (anticancer drug MK-2206) and interaction with ECM. The spheroids present apical plasma membrane on the surface, which originated from the failure of polarisation at the single-cell stage and subsequent defects in phosphorylated ezrin accumulation at the cell boundary during the first cleavage, failing internal lumen formation. At the multicellular stage, liquid culture spheroids presented bleb-like protrusion on the surface, which was enhanced by the activation of the PI3K/Akt pathway and reduced by PI3K/Akt inhibitors. Liquid culture spheroids exhibited slow proliferation speed and low endogenous pAkt levels compared with gel-cultured spheroids and 2D-cultured cells, explaining the susceptibility to the Akt-inhibiting anticancer drug. Subcutaneous xenografting and in vitro analysis demonstrated low viability and adhesive property of liquid culture spheroids to ECM, while migratory and invasive capacities were comparable with gel-cultured spheroids. These features agree with the low efficacy of circulating tumour spheroids in the settling step of metastasis. This study demonstrates the feature of anchorage-independent spheroids and validates liquid cultures as a useful method in cancer spheroid research.

Utility of human papillomavirus L1 capsid protein and HPV test as prognostic markers for cervical intraepithelial neoplasia 2+ in women with persistent ASCUS /LSIL cervical cytology.

Objective: Efficient and highly predictive biomarkers reflecting the prognosis of persistent atypical squamous cells of unknown significance(ASCUS) and low grade squamous intraepithelial lesion(LSIL)s are unavailable and need to be developed urgently. We aimed to develop a predictive model for diagnosis of cervical intraepithelial neoplasia(CIN)2+ by analyzing the immunocytochemical expression of the HPV L1 capsid protein in patients with persistent ASCUS and LSIL with a high risk of HPV infection. Methods: Cervical cytology samples comprising (70 ASCUS and 215 LSIL Pap smears) were analyzed. Immunocytochemical identification of the HPV L1 capsid protein in cervical cytology samples was performed. Expression levels of HPV L1 capsid protein in cervical cytology samples were measured, and the correlation between HPV L1 expression and cervical pathologic diagnosis was evaluated. The risk for CIN2+ was calculated using the results of immunocytochemistry and the HPV DNA test. Results: Negative results for HPV L1 immunochemistry test were more frequently observed in CIN2+, and expression of the HPV L1 capsid protein was higher in CIN1 or cervicitis (Fisher's exact test, p<0.05). Diagnosis rates for CIN2+ were highest for the combination of HPV L1 capsid protein immunocytochemistry, cytology and HPV test when compared with other combinations (Akaike information criterion (AIC): 191.7, Schwarz criterion(SC): 206.3, p<0.001). Conclusion: Absence of HPV L1 capsid expression and presence of HPV type 16 or 18 infection are reliable predictors of progression to CIN2+ in patients showing persistent ASCUS and LSIL.

Differences in autophagy-associated mRNAs in peritoneal fluid of patients with endometriosis and gynecologic cancers.

Endometriosis and gynecologic cancer show similar patterns of invasion. Little is known about the roles of autophagy in endometriosis and, to date, the expression of autophagy-associated mRNAs has not been compared in patients with endometriosis and gynecologic cancers. This study therefore compared the levels of expression of autophagy-associated mRNAs in patients with endometriosis and gynecologic cancers. The levels of autophagy mRNAs, including those encoding mTOR, P13KC3, Beclin-1, Bcl-2, LC3 II, FLIP, Rubicon, BIRC2 and BIRC5, were measured by real time polymerase chain reaction in peritoneal fluid of 27 patients with benign masses (control group), 42 patients with endometriosis, and 43 patients with gynecologic (ovarian, uterine, and cervical) cancers. Findings in the three groups were compared. Autophagy mRNAs were present in all samples from patients with endometriosis and gynecologic cancers. The levels of PI3K, FLIP, and Rubicon mRNAs were significantly higher in the endometriosis than in the control group (p < 0.05 each). Compared with the gynecologic cancer group, the levels of LC3II and FLIP mRNAs were significantly lower, and the levels of Beclin-1 and Rubicon mRNAs significantly higher, in the endometriosis group (p < 0.05 each). Levels of PI3K and FLIP mRNA were significantly higher in the endometriosis and gynecologic cancer groups than in the control group (p < 0.05 each). PI3K, FLIP, and Rubicon mRNAs are closely associated with the pathogenesis of endometriosis. The similar increases in PI3K and FLIP mRNA expression observed in patients with endometriosis and gynecologic cancer suggest that these conditions have similar autophagic characteristics. The lower levels of Beclin-1 mRNA in the gynecologic cancer than in other two groups suggest that lower Beclin-1 mRNA levels increase the likelihood of developing gynecologic cancer.

Cryoprotective effect of antifreeze polyamino-acid (Carboxylated Poly-l-Lysine) on bovine sperm: A technical note.

In this study, investigating Carboxylated Poly-l-Lysine (CPLL) for its effectiveness as a new cryoprotectant for bovine sperm is aimed. CPLL is an ampholytic polymer compound, has cryoprotective properties similar to those of anti-freeze protein. The cryopreservation medium used for control group consisted of 6.5% (v/v) glycerol, the cryopreservation medium used for experimental group consisted of 3.25% (v/v) glycerol + 0.5% (w/v) CPLL. There was no consequential difference in sperm motility parameter after thawing whereas there was huge distinction for sperm membrane integrity rate (control vs experimental; 49.6 vs 60.7%, P < 0.01). Conception rate of artificial insemination of experimental group was significantly higher than that of control group (79.0% vs 53.1%, P < 0.01). These results suggest CPLL has protected sperm membrane and leads to improve fertility. This is the first report using CPLL for bovine sperm cryopreservation, it is also expected CPLL can be applied to other animal species.

Cryopreservation of bovine somatic cells using antifreeze polyamino-acid (carboxylated poly-L-lysine).

Carboxylated poly-L-lysine (CPLL) is an ampholytic polymer compound, obtained by converting 65 mol% of amino groups to carboxyl groups after synthesizing ε-poly-l-lysine aqueous solution and succinic anhydride. CPLL has cryoprotective properties similar to those of anti-freeze protein. The addition of CPLL to freezing medium has been reported to improve the post-thawing survival rate of murine cells, human induced pluripotent stem (iPS) cells, embryonic stem (ES) cells and embryos. In this study, investigating CPLL for its effectiveness as a new cryoprotective material is aimed. In experiments with bovine somatic cells, CPLL was suggested to have an equal or superior cryoprotective effect to dimethyl sulfoxide (DMSO), the conventional material for cellular frozen storage, based on the results for post-thawing cell survival and proliferation rates. CPLL was demonstrated to have another advantage; thawed cells can be cultured without removing the cryopreservation medium when CPLL is used, but not when DMSO is used. These results suggest that CPLL could be used as cryoprotective material for bovine cells. It is also expected that CPLL can be applied to embryo and oocytes storage for cattle, and similar functions for cells and embryos of other animal species.

Expression of Cytokeratin 13, 14, 17, and 19 in 4-nitroquinoline-1-oxide-induced Oral Carcinogenesis in Rat.

The management of epithelial dysplastic spread around an oral squamous cell carci-noma is very important, particularly intraoperatively. Both cytokeratin (CK) 14 and CK19 are believed to be involved in the development of precancerous lesions, and their expression profiles are quite specific in these and early cancer lesions. Here, expression of CK13, 14, 17, and 19 was investigated in a rat model of 4-nitroquinoline-1-oxide-induced tongue cancer during a series of carcinogenetic processes to determine their value in assessing the features of epithelial dysplastic spread around a cancer. Based on tissue conditions, the results showed that expression levels of CK13 and 14 decreased in the order of no change, dysplasia, and cancer, whereas those of CK17 and 19 increased in the same order. Expression of CK13 showed a significant difference among no change, dysplasia, and cancer. This indicates that comparing the immunohistochemical staining profiles of CKs, especially CK13, could help in assessing the characteristics of epithelial dysplastic spread around a cancer.

Expression of Cytokeratin 14 and 19 in Process of Oral Carcinogenesis.

Cytokeratins (CK) are abundant in keratinized cells, particularly CK14 and CK19, which are expressed in stratified squamous epithelial cells. In this study, expression of CK14 and 19 was examined in human epithelial and dysplastic tissues. Surgical specimens from patients with clinically diagnosed oral leukoplakia or early cancer were stained with hematoxylin and eosin and classified into normal, low grade dysplasia (LGD), high grade dysplasia (HGD), or squamous cell carcinoma (SCC). The sections were examined by immunostaining and reverse transcription-polymerase chain reaction (RT-PCR) for CK14 and CK19. Expression and the results of RT-PCR for CK14 showed a decrease in the order of LGD, HGD, and SCC, whereas those of CK19 showed an increase in that order. These results suggest that decreased expression of CK14 and increased expression of CK19 serve as indicators of potential for malignant transformation.

Evaluation of postextraction bleeding incidence to compare patients receiving and not receiving warfarin therapy: a cross-sectional, multicentre, observational study.

OBJECTIVES: We investigated incidence and risk factors for postextraction bleeding in patients receiving warfarin and those not receiving anticoagulation therapy. DESIGN: Cross-sectional, multicentre, observational study. SETTING: 26 hospitals where an oral surgeon is available. PARTICIPANTS: Data on 2817 teeth (from 496 patients receiving warfarin, 2321 patients not receiving warfarin; mean age (SD): 62.2 (17.6)) extracted between 1 November 2008 and 31 March 2010, were collected. Warfarin-receiving patients were eligible when prothrombin time-international normalised ratio (PT-INR) measured within 7 days prior to the extraction was less than 3.0. INTERVENTIONS: Simple dental extraction was performed, and incidence of postextraction bleeding and comorbidities were recorded. PRIMARY AND SECONDARY OUTCOME MEASURES: Postextraction bleeding not controlled by basic haemostasis procedure was clinically significant. RESULTS: Bleeding events were reported for 35 (7.1%) and 49 (2.1%) teeth, of which 18 (3.6%) and 9 (0.4%) teeth were considered clinically significant, in warfarin and non-warfarin groups, respectively, the difference between which was 3.24% (CI 1.58% to 4.90%). The incidence rates by patients were 2.77% and 0.39%, in warfarin and non-warfarin groups, respectively (incidence difference 2.38%, CI 0.65% to 4/10%). Univariate analyses showed that age (OR 0.197, p=0.001), PT-INR (OR 3.635, p=0.003), mandibular foramen conduction anaesthesia (OR 4.854, p=0.050) and formation of abnormal granulation tissue in extraction socket (OR 2.900, p=0.031) significantly correlate with bleeding incidence. Multivariate analysis revealed that age (OR 0.126, p=0.001), antiplatelet drugs (OR 0.100, p=0.049), PT-INR (OR 7.797, p=0.001) and history of acute inflammation at extraction site (OR 3.722, p=0.037) were significant risk factors for postextraction bleeding. CONCLUSIONS: Our results suggest that there is slight but significant increase in the incidences of postextraction bleeding in patients receiving warfarin. Although absolute incidence was low in both groups, the bleeding risk is not negligible.

Ventilator-associated pneumonia risk decreased by use of oral moisture gel in oral health care.

Although oral health care has a preventive effect against ventilator-associated pneumonia (VAP), the most effective method of oral health care in this respect remains to be established. The objective of this single-center, randomized, controlled trial was to investigate the relationship between VAP and various methods of oral health care. All patients included in the study (n=142) were on mechanical ventilation with oral intubation at the intensive care unit of the Tokyo Dental College Ichikawa General Hospital. They were divided into two groups, one receiving standard oral health care (Standard group), and the other receiving oral health care using an oral moisture gel instead of water (Gel group). After removal of the intubation tube, biofilm on cuff of the tube was stained with a disclosing agent to determine the contamination level. Factors investigated included sex, age, number of remaining teeth, intubation time, fever ≥38.5°C, VAP, cuff contamination level, and time required for one oral health care session. No VAP occurred in either group during the study period. The level of cuff contamination was significantly lower in the Gel group than the Standard group, and the time required for one session of oral health care was shorter (p<0.001). Multivariate analysis revealed use of the oral moisture gel as a factor affecting cuff contamination level. Use of an oral moisture gel decreased invasion of the pharynx by bacteria and contaminants together with biofilm formation on the intubation tube cuff. These results suggest that oral health care using an oral moisture gel is effective in preventing cuff contamination.

Evaluation of superficial oral squamous cell malignancy based on morphometry and immunoexpression of cytokeratin 13 and cytokeratin 17.

OBJECTIVE: Evaluation of combined morphometry and immunoexpression of cytokeratin 13 (CK13) and cytokeratin 17 (CK17) for cytological identification of superficial oral squamous cells. STUDY DESIGN: Smears from 11 tongue squamous cell carcinoma patients were processed by liquid-based cytology, stained via the Papanicolaou method and divided into multiple specimens by cell transfer. Morphometric indices, including nuclear area, nuclear perimeter, nuclear circular rate, largest-to-smallest dimension ratio of the nucleus and nucleocytoplasmic ratio, were measured using a computerized analysis system. CK13 and CK17 were detected by immunostaining. Morphometric values were compared between cell populations with distinct staining and immunoexpression patterns. RESULTS: Most orange G-stained superficial cells were negative for CK13 (99.4%) and CK17 (98.6%). For light green-stained superficial cells, loss of CK13 was associated with greater cellular atypia in the nuclear area, nuclear perimeter and nucleocytoplasmic ratio (p < 0.01), while expression of CK17 was related to higher-grade cellular atypia in the same parameters (p < 0.01) as well as the nuclear circular rate (p < 0.05). CONCLUSION: Immunoexpression of CK13 and CK17 in light green-stained superficial cells was associated with more severe morphological atypia. Combined morphometry and immunoexpression of CK13 and CK17 might be useful for cytological diagnosis of this cell population.

Re-epithelialization of the Buccal Mucosa after Alkaline Chemical Injury.

Alkaline conditions in the oral cavity may be caused by a variety of stimuli, including tobacco products, antacids, alkaline drinking water and bicarbonate toothpaste. However, the effects of an alkaline pH on the oral mucosa had not been elucidated. The purpose of this study was to investigate how basal keratinocytes are actively involved in re-epithelialization after alkaline chemical injury. We generated epithelial defects in the oral mucosa of mice by applying an alkaline chemical, and the localization of cytokeratin 13, cytokeratin 14, PCNA and p63 was investigated during the re-epithelialization process. PCNA- and p63-positive staining was seen in basal cells covering the wound surface at 1 day after the chemical injury. Cytokeratin 14-positive and PCNA-negative basal keratinocytes were localized in a few layers of the wound epithelium during epithelial outgrowth. Cytokeratin 14-positive and PCNA-positive basal keratinocytes, indicating proliferation, were localized over the entire layer of the epithelium at the wound margin. These results imply that basal keratinocytes at the wound margin migrate to the wound surface, provoke differentiation and keratinization during epithelial outgrowth and that epithelial cells are supplied from the wound margin to the epithelial outgrowth after alkaline chemical injury.

Head and neck contrast-enhanced CT for identification of internal carotid artery stenosis progression on the affected side after treatment for oral squamous cell carcinoma.

OBJECTIVES: To determine the incidence of progressive internal carotid artery (ICA) stenosis by head and neck contrast-enhanced computed tomography (CT) in 82 patients who underwent surgery, chemotherapy, or combination therapy for oral squamous cell carcinoma (OSCC). METHODS: The study included 82 patients who underwent head and neck contrast-enhanced CT after surgery alone or combined surgery and chemotherapy for OSCC at the Department of Oral and Maxillofacial Surgery of Ichikawa General Hospital, Tokyo Dental College, or Tokyo Dental College Oral Cancer Center between December 2002 and March 2010. RESULTS: Comparison with previously obtained head and neck contrast-enhanced CT images revealed progressive arterial stenosis of the ICA in five patients with a mean age of 62.0 years. All five patients were male, and their OSCC sites were the tongue in two, the floor of the mouth in two, and the mandibular gingiva in one. Tumor resection and neck dissection were performed for four patients and tumor resection alone for one patient. Four patients underwent chemotherapy. ICA stenosis occurred on the same side as the tumor in all five patients. CONCLUSIONS: The results of this study suggest that, given the possibility of post-treatment vascular events, attention must be paid to subsequent changes in the ICA over time. The results also indicate the usefulness of head and neck contrast-enhanced CT in identifying such problems.

Expression of myelin and lymphocyte protein (MAL) in oral carcinogenesis.

In the pathological diagnosis of oral squamous cell carcinoma, we often confront the difficulty of determining whether it is invasive carcinoma or epithelial dysplasia. Recently, myelin and lymphocyte protein (MAL; T-cell differentiation-related gene) has been reported to be a candidate gene suppressed in esophageal carcinoma. When we performed cDNA microarray analysis, we found that gene expression of MAL was significantly downregulated in oral squamous cell carcinoma (OSCC). We evaluated the expression of the MAL gene by laser microdissection and real-time PCR methods and protein localization by immunohistochemistry. The gene expression of MAL was significantly decreased in OSCC compared with normal epithelium (P < 0.05). Furthermore, protein expression of MAL disappeared gradually in proportion to malignancy. The results suggest that MAL plays an important role during oral carcinogenesis and that the gene may have potential as a biomarker target for OSCC.

Osteogenic differences in cultured rat periosteal cells under hypoxic and normal conditions.

The aim of the present study was to investigate the osteogenic capability of rat calvarial periosteal cells in hypoxic conditions in vitro. Periosteum was obtained from the calvarial bone of Sprague-Dawley rats. Following primary tissue culture, subcultured cells were used in hypoxic or normal conditions. On days 1, 2, 3 and 4 following the cell culture, cell proliferation and mRNA and protein expression levels were evaluated. No significant difference in the cell proliferation rate was found between the normal and hypoxic condition groups. The hypoxic condition group exhibited a stronger expression of hypoxia-inducible factor (HIF)1α, vascular endothelial growth factor (VEGF), Runx2, alkaline phosphatase (ALP), bone sialoprotein (BSP), osteocalcin (OCN) and periostin at the mRNA level compared to that of the normal condition group. The hypoxic condition group also exhibited a stronger expression of HIF1α, VEGF, bone morphogenetic protein (BMP)2, Runx2, ALP and BSP at the protein level compared to that of the normal condition group. In conclusion, periosteal cells cultured in hypoxic conditions demonstrated activated osteogenic capability in vitro.

Risk factors for postoperative dysphagia in oral cancer.

With the founding of its Oral Cancer Center at the Ichikawa General Hospital, Tokyo Dental College established a support system for patients and family members that not only provides surgery and other conventional cancer-oriented treatments, but also palliative care, nutritional support, rehabilitation, and discharge support. With this in mind, the present study sought to examine the nature of support for oral cancer patients with postoperative eating and swallowing disorders by investigating these disorders and identifying their risk factors. The study population comprised 75 surviving oral cancer patients (46 men and 29 women) discharged from the Tokyo Dental College Oral Cancer Center following treatment over a 2-year period from April 2009 to March 2011. Risk factors affecting eating and swallowing function were identified by statistical analysis. Mean age of the patients was 67.3±13.7 years. Fifteen patients had stage I cancer, while 25 had stage II, 13 had stage III, and 22 had stage IV. The feeding route at the time of discharge was oral feeding in 74 patients and a combination of oral and gastrostomy tube feeding in 1 patient. The Tokyo Dental College Ichikawa General Hospital has standardized the expert evaluation and rehabilitation of oral cancer patients with eating and swallowing disorders by establishing a multidisciplinary support system from the preoperative stage onwards. In this context, the results of our analysis of factors influencing the ability of oral cancer patients to orally ingest food after treatment suggest that preoperative cancer stage classification, neck dissection, and tracheotomy are all influential factors. Patients affected by these factors require further multidisciplinary treatment, which in turn necessitates more extensive coordination with other medical professionals and community health care providers.