Tamoxifen Alters TGF-ß1/Smad Signaling in Vocal Fold Injury.

OBJECTIVES: Effective treatments for vocal fold fibrosis remain elusive. Tamoxifen (TAM) is a selective estrogen receptor modulator and was recently reported to have antifibrotic actions. We hypothesized that TAM inhibits vocal fold fibrosis via altered transforming growth factor beta 1 (TGF-ß1) signaling. Both in vitro and in vivo approaches were employed to address this hypothesis. METHODS: In vitro, vocal fold fibroblasts were treated with TAM (10-8 or 10-9 M) ± TGF-ß1 (10 ng/ml) to quantify cell proliferation. The effects of TAM on genes related to fibrosis were quantified via quantitative real-time polymerase chain reaction. In vivo, rat vocal folds were unilaterally injured, and TAM was administered by oral gavage from pre-injury day 5 to post-injury day 7. The rats were randomized into two groups: 0 mg/kg/day (sham) and 50 mg/kg/day (TAM). Histological changes were examined on day 56 to assess tissue architecture. RESULTS: TAM (10-8 M) did not affect Smad3, Smad7, Acta2, or genes related to extracellular matrix metabolism. TAM (10-8 or 10-9 M) + TGF-ß1, however, significantly increased Smad7 and Has3 expression and decreased Col1a1 and Acta2 expression compared to TGF-ß1 alone. In vivo, TAM significantly increased lamina propria area, hyaluronic acid concentration, and reduced collagen deposition compared to sham treatment. CONCLUSIONS: TAM has antifibrotic potential via the regulation of TGF-ß1/Smad signaling in vocal fold injury. These findings provide foundational data to develop innovative therapeutic options for vocal fold fibrosis. LEVEL OF EVIDENCE: NA Laryngoscope, 133:2248-2254, 2023.

Effects of Repeated Intracordal Glucocorticoid Injection on the Histology and Gene Expression of Rat Vocal Folds.

OBJECTIVES: Local injection of glucocorticoids (GCs) into the vocal folds has been used for treating the vocal fold lesions. While the positive effects on vocal fold nodules, polyps, or scarring have been clinically reported, some concern remains around the potential adverse effects such as vocal fold atrophy, and the mechanisms remain unclear. The present study examined the histology and gene expression of locally injected GC into the vocal folds in rats. METHODS: Thirteen-week-old male Sprague-Dawley rats were used in the experiments. Triamcinolone acetonide (TAA) or saline were administered repeatedly to the right vocal folds at a weekly interval, and rats were euthanized one week after the last administration for histological examination. Genetic examination was assessed hyaluronic acid (HA) metabolism at 1 or 3 days after a single TAA injection by quantitative real-time polymerase chain reaction (qRT-PCR). RESULTS: The group which underwent four TAA injections showed a significant decrease in HA in the lamina propria (LP), thickness of the LP and total cell numbers of the LP compared with the saline group. In contrast, there was no significant difference in the area of collagen accumulation and the thyroarytenoid muscle, although there was a tendency of atrophy of the muscle. After single injection of TAA, qRT-PCR showed a significant decrease in the expression of HA synthases, Has2 and Has3. CONCLUSIONS: The current animal study first demonstrates that repeated intracordal injection of GCs may lead to atrophy of vocal folds caused by decrease of deposition of HA in the LP and decrease of gene expression of Has.

Thymidylate synthase locus LOH in combination with genotype has prognostic and predictive significance in colorectal cancer.

The aim of the current study was to investigate the prognostic and predictive significance of polymorphisms in the thymidylate synthase (TS) gene, alongside the loss of heterozygocity (LOH) at this gene locus in patients with colorectal cancer. Genotyping was carried out for a variable number tandem repeat (VNTR) polymorphism in the TS 5'-untranslated region, a G/C single nucleotide polymorphism (SNP) located within this VNTR, and for TS LOH status in 246 colorectal cancer and paired normal DNA samples. The results were analyzed in relation to clinicopathological features, including the prognostic and predictive significance of TS genotype in patients who underwent curative surgery. Complete VNTR, SNP and LOH information for TS was obtained in 226 cases. No significant associations were observed between normal tissue TS genotype status and clinicopathological features. LOH of TS was observed in 58% of tumor samples and was associated with poor prognosis independently of clinical stage. Cases exhibiting TS LOH were classified into the three groups of 2R/loss, 3G/loss and 3C/loss. Patients with 3C/loss genotype status had poor outcomes when treated by surgery alone, but their survival was similar to patients with other genotypes following Fluorouracil (5-FU)-based adjuvant chemotherapy. The results suggested that LOH of the TS locus may be a significant prognostic factor in colorectal cancer, with the genotype of the residual allele also demonstrating an influence on prognosis. In conclusion, LOH status should be considered when TS genotype is explored as a potential prognostic and predictive marker for 5-FU-based adjuvant chemotherapy in colorectal cancer.

Voice rest after laryngeal surgery: what's the evidence?

PURPOSE OF REVIEW: This article aims to review previous research reports and to summarize current strategies for the optimal duration of voice rest and the effect of phonatory stimulation after phonomicrosurgery. RECENT FINDINGS: Voice rest is commonly recommended after laryngeal surgery to prevent worsening of vocal fold injuries. However, there are no established standard protocol for voice rest, and the type and duration of voice rest vary among clinicians. The most effective duration of voice rest is unknown. Recently, early vocal stimulation was recommended as a means to improve wound healing, on the basis of the basic and clinical researches. SUMMARY: It seems that early vocal stimulation may enhance the wound healing process in the vocal fold. More basic and clinical researches are warranted to investigate appropriate timing of initiation of stimulation, as well as the type and amount of stimulation that are available for human.

Enhancement of nose-to-brain delivery of hydrophilic macromolecules with stearate- or polyethylene glycol-modified arginine-rich peptide.

Recently, nose-to-brain delivery is a highly versatile route, which, in combination with novel drugs being developed for treating intractable CNS diseases, is a promising approach for the treatment of disorders. Furthermore, nano-sized drug carriers may improve nose-to-brain drug delivery by their capability to increase the transmucosal penetration of the drugs across nasal mucosal tissue barrier. However, there is still not enough information regarding mechanism of absorption pathway from nasal cavity to brain using nanocarriers. In this study, to investigate the nose-to-brain transport pathway using nanocarriers, the distribution in whole brain, nasal mucosa, and trigeminal nerve after intranasal administration of two kinds of nanocarriers which have hydrophobic or hydrophilic moiety. We used CHHRRRRHHC peptide (CH2R4H2C) as basic peptide carriers, and modified with stearic acid (STR) as a hydrophobic moiety (STR-CH2R4H2C) or polyethylene glycol (PEG)-based block copolymer (PEG-PCL) as hydrophilic moiety (PEG-PCL-CH2R4H2C). The nose-to-brain drug delivery can be improved by using STR-CH2R4H2C and PEG-PCL-CH2R4H2C as carriers. Specifically, hydrophobic STR-CH2R4H2C is more suitable for the transport of drugs targeting the forebrain, while PEG-PCL-modified CH2R4H2C is more suitable for transporting drugs targeting the hindbrain or whole brain tissue. In conclusion, the results of this study support the possibility that drug delivery pathways can be controlled depending on the properties of different carrier complexes.

Optimal Duration for Voice Rest After Vocal Fold Surgery: Randomized Controlled Clinical Study.

OBJECTIVES: Voice rest is commonly recommended after phonomicrosurgery to prevent worsening of vocal fold injuries. However, the most effective duration of voice rest is unknown. Recently, early vocal stimulation was recommended as a means to improve wound healing. The purpose of this study is to examine the optimal duration of voice rest after phonomicrosurgery. STUDY DESIGN: Randomized controlled clinical study. METHODS: Patients undergoing phonomicrosurgery for leukoplakia, carcinoma in situ, vocal fold polyp, Reinke's edema, and cyst were chosen. Participants were randomly assigned to voice rest for 3 or 7 postoperative days. Voice therapy was administered to both groups after voice rest. Grade, roughness, breathiness, asthenia, and strain (GRBAS) scale, stroboscopic examination, aerodynamic assessment, acoustic analysis, and Voice Handicap Index-10 (VHI-10) were performed pre- and postoperatively at 1, 3, and 6 months. Stroboscopic examination evaluated normalized mucosal wave amplitude (NMWA). Parameters were compared between both groups. RESULTS: Thirty-one patients were analyzed (3-day group, n = 16; 7-day group, n = 15). Jitter, shimmer, and VHI-10 were significantly better in the 3-day group at 1 month post operation. GRBAS was significantly better in the 3-day group at 1 and 3 months post operation, and NMWA was significantly better in the 3-day group at 1, 3, and 6 months post operation compared to the 7-day group. CONCLUSIONS: The data suggest that 3 days of voice rest followed by voice therapy may lead to better wound healing of the vocal fold compared to 7 days of voice rest. Appropriate mechanical stimulation during early stages of vocal fold wound healing may lead to favorable functional recovery.

Prognostic and predictive significance of long interspersed nucleotide element-1 methylation in advanced-stage colorectal cancer.

BACKGROUND: Hypomethylation of Long Interspersed Nucleotide Element-1 (LINE-1) is associated with worse prognosis in colorectal cancer (CRC). However, little is known about the relevance of this marker for the prognosis and response to chemotherapy of metastatic and recurrent (advanced-stage) CRC. Our aim was therefore to investigate whether tumor LINE-1 hypomethylation correlates with patient survival and with response to 5-fluorouracil (5-FU)/ oxaliplatin (FOLFOX) chemotherapy in advanced-stage CRC. METHODS: The study included 40 CRC patients who developed metastasis or local recurrence after surgery and subsequently underwent FOLFOX therapy. Progression-free and overall survival were estimated using the Kaplan-Meier method. LINE-1 methylation levels in formalin-fixed and paraffin-embedded primary tumor tissues were measured by MethyLight assay and correlated with patient survival. In vitro analyses were also conducted with human colon cancer cell lines having different LINE-1 methylation levels to examine the effects of 5-FU and oxaliplatin on LINE-1 activity and DNA double-strand-breaks. RESULTS: Patients with LINE-1 hypomethylation showed significantly worse progression-free (median: 6.6 vs 9.4 months; P = 0.02) and overall (median: 16.6 vs 23.2 months; P = 0.01) survival following chemotherapy compared to patients with high methylation. LINE-1 hypomethylation was an independent factor for poor prognosis (P = 0.018) and was associated with a trend for non-response to FOLFOX chemotherapy. In vitro analysis showed that oxaliplatin increased the LINE-1 score in LINE-1-expressing (hypomethylated) cancer cells, thereby enhancing and prolonging the effect of 5-FU against these cells. This finding supports the observed correlation between tumor LINE-1 methylation and response to chemotherapy in CRC patients. CONCLUSIONS: Tumor LINE-1 hypomethylation is an independent marker of poor prognosis in advanced-stage CRC and may also predict non-response to combination FOLFOX chemotherapy. Prospective studies are needed to optimize the measurement of tumor LINE-1 methylation and to confirm its clinical impact, particularly as a predictive marker.

Construction of a tissue-specific transcription factor-tethered extracellular matrix protein via coiled-coil helix formation.

Tissue-specific transcription factors are key regulators of cellular differentiation. Previously, we succeeded in introducing basic helix-loop-helix tissue-specific transcription factor proteins into cells to induce cellular differentiation. Based on these results, we decided to focus on the use of tissue-specific transcription factor proteins in the construction of biomaterials. In this proof-of-concept study, we demonstrate the construction of a tissue-specific transcription factor-tethered extracellular matrix protein. Here, the tissue-specific transcription factor Olig2 was tethered to a designed artificial extracellular matrix protein via coiled-coil helix formation. Tethered Olig2 was introduced into mouse embryonic carcinoma P19 cells attached to our designed extracellular matrix protein, and was shown to exhibit the ability to induce neural differentiation.

Comparison of vocal outcomes after angiolytic laser surgery and microflap surgery for vocal polyps.

OBJECTIVE: The microflap technique is a standard procedure for the treatment of vocal fold polyps. Angiolytic laser surgery carried out under topical anesthesia is an alternative method for vocal polyp removal. However, it is not clear whether angiolytic laser surgery has the same effects on vocal outcomes as the microflap technique because of a lack of studies comparing both procedures. In the current study, vocal outcomes after both procedures were compared to clarify the effects of angiolytic laser surgery for vocal polyp removal. METHODS: Vocal outcomes were reviewed for patients who underwent angiolytic laser surgery (n=20, laser group) or microflap surgery (n=34, microflap group) for vocal polyp removal. The data analyzed included patient and lesion characteristics, number of surgeries required for complete resolution, and aerodynamic and acoustic examinations before and after surgery. RESULTS: In the laser surgery group, complete resolution of the lesion was achieved with a single procedure in 17 cases (85%) and with two procedures in 3 cases (15%). Postoperative aerodynamic and acoustic parameters demonstrated significant improvement compared to preoperative parameters in both the laser surgery group and the microflap surgery group. There were no significant differences in any postoperative aerodynamic and acoustic parameters between the two groups. CONCLUSION: The current retrospective study demonstrated that angiolytic laser surgery achieved complete resolution of vocal polyps within two procedures. Postoperative effects on aerodynamic and acoustic functions were similar to those after microflap surgery.

Multidimensional Analysis on the Effect of Vocal Function Exercises on Aged Vocal Fold Atrophy.

OBJECTIVES: Age-related voice change is characterized as weak, harsh, and breathy. These changes are caused by histologic alteration of the lamina propria of the vocal fold mucosa as well as atrophy of the thyroarytenoid muscle. Several therapeutic strategies involving laryngeal framework surgery and injection laryngoplasty have been tried, but effects have been limited. Vocal function exercises (VFE) have been used to treat age-related vocal fold atrophy although the effectiveness has been shown with limited analysis. The present study aims to determine the effectiveness of VFE for the treatment of aged atrophy using multidimensional analysis. STUDY DESIGN: This is a retrospective study. METHODS: Sixteen patients with vocal fold atrophy aged 65-81 years underwent voice therapy using VFE. Six patients with vocal fold atrophy aged 65-85 years were involved as a historical control group. The grade, roughness, breathiness, asthenia, strain (GRBAS) scale, stroboscopic examinations, aerodynamic assessment, acoustic analysis, and Voice Handicap Index-10 (VHI-10) were performed before and after VFE. Normalized mucosal wave amplitude (NMWA), normalized glottal gap (NGG), and bowing index (BI) were measured by image analysis during stroboscopic examinations. RESULTS: After VFE, significant improvements were shown in GRBAS, maximum phonation time, jitter, NMWA, NGG, and VHI-10 although BI has not changed significantly. There were no significant improvements in the historical control. CONCLUSIONS: The data suggest that VFE produces significant improvement in subjective, objective, and patient self-evaluation and deserves further attention as a treatment for aged atrophy of the vocal fold. It was also suggested that VFE does not improve the vocal fold bowing but may improve muscular function during voicing.

Chitosan tubes can restore the function of resected phrenic nerves.

OBJECTIVES: We previously reported that the phrenic nerve could be morphologically repaired by implantation of a chitosan nanofibre tube (C-tube). In the current study, we investigated whether implantation of C-tubes could improve the function of an injured phrenic nerve using a beagle dog model. METHODS: Seven beagle dogs underwent right thoracotomy under general anaesthesia. An approximately 5 mm length of the right phrenic nerve was resected. Five dogs had a C-tube implantation (C-tube group) and other two dogs did not have the C-tube implantation (control group). Diaphragm movements were longitudinally measured by X-ray fluoroscopy before surgery, immediately after the surgery, and 3, 6 and 12 months after the surgery. The diaphragm movement was determined by diaphragm levels at inspiration and expiration phases, and the excursion difference between them was calculated. At 12 months after the surgery, rethoracotomy was performed to examine electrical phrenic nerve conduction. The C-tube and phrenic nerve were then excised for histological assessment of nerve regeneration. RESULTS: Three of the five animals of the C-tube group showed improvement of diaphragm movement with time. In these three animals, slow phrenic nerve conduction was observed. Histological assessment showed that the injured nerve was connected by newly regenerating nerve fibres surrounded by granulation tissue within the C-tube. On the other hand, the animals in the control group and two animals of the C-tube group showed neither improved diaphragm movement, nor electrical conduction to the diaphragm. No nerve fibre regeneration was found by histology. CONCLUSIONS: Our results suggest that, in addition to morphological improvement, C-tube implantation can functionally improve the injured phrenic nerve by promoting phrenic nerve regeneration.

Regenerative phonosurgical treatments for vocal fold scar and sulcus with basic fibroblast growth factor.

OBJECTIVES/HYPOTHESIS: Vocal fold scar and sulcus are still challenges. Basic fibroblast growth factor (bFGF) has proven to be effective to resolve scar tissue in animal models. This study reports the efficacy of regenerative treatments using bFGF on vocal fold scar and sulcus in human cases. STUDY DESIGN: Retrospective chart review. METHODS: Fifteen cases (7 scar; 8 sulcus) were treated by either local injection of bFGF (n = 6) or regenerative surgery using bFGF (n = 9). Injection regimen was to locally apply 10 micrograms of bFGF in 0.5 mL saline into each vocal fold under topical anesthesia repeatedly (4 times with intervals of 1 week between each injection). The regenerative surgical procedure consisted of the dissection of scar tissue and the implant of gelatin sponge with bFGF. Follow-up periods ranged from 6 months to 24 months. RESULTS: Maximum Phonation Time (MPT); Voice Handicap Index (VHI)-10; and Grade, Roughness, Breathiness, Asthenia, Strain (GRBAS) scale were assessed in both groups. The injection group showed significant improvement on VHI-10 and GRBAS. The regenerative surgery group showed significant improvement in all parameters. Jitter and shimmer were evaluated in the surgery group, and the results indicated improvement in six and five cases of nine cases, respectively. No major adverse effects were observed in both treatment groups. CONCLUSIONS: Regenerative treatments using bFGF has shown to be effective for improvement of vocal function in scar and sulcus.

[A case of recurrent breast cancer with multiple bone metastasis successfully treated with S-1 and zoledronic acid therapy].

For a case of recurrent breast cancer with multiple bone metastasis, an oral pyrimidine fluoride-based anti-cancer drug S -1, and zoledronic acid(a third-generation bisphosphonate formulation), were prescribed in experiments to test their efficacy. S-1 was prescribed orally at doses of 100 mg/day(twice)over a 4-week period with a cessation period of 2 weeks. Zoledronic acid was commenced with a strict administration of 4 mg every 4 weeks, taken intravenously. At the conclusion of the 1st S-1 course, the tumor markers began to improve to a certain extent with an improvement in the symptoms. Following the 16 courses, there was no abnormal accumulation detected by PET. Now, at the end of the 21st course, the treatment is being continued, and there has been no recurrence of inflammation or reoccurring growth detected. No adverse effects of more than Grade 1 occurred during the elapsed process. As a form of combined therapy, we can expect zoledronic acid to be a good anti-cancer drug because of its effectiveness and tolerability in treating recurring breast cancer.

LINE-1 methylation shows little intra-patient heterogeneity in primary and synchronous metastatic colorectal cancer.

BACKGROUND: Long interspersed nucleotide element 1 (LINE-1) hypomethylation is suggested to play a role in the progression of colorectal cancer (CRC). To assess intra-patient heterogeneity of LINE-1 methylation in CRC and to understand its biological relevance in invasion and metastasis, we evaluated the LINE-1 methylation at multiple tumor sites. In addition, the influence of stromal cell content on the measurement of LINE-1 methylation in tumor tissue was analyzed. METHODS: Formalin-fixed paraffin-embedded primary tumor tissue was obtained from 48 CRC patients. Matched adjacent normal colon tissue, lymph node metastases and distant metastases were obtained from 12, 18 and 7 of these patients, respectively. Three different areas were microdissected from each primary tumor and included the tumor center and invasive front. Normal mucosal and stromal cells were also microdissected for comparison with the tumor cells. The microdissected samples were compared in LINE-1 methylation level measured by multicolor MethyLight assay. The assay results were also compared between microdissected and macrodissected tissue samples. RESULTS: LINE-1 methylation within primary tumors showed no significant intra-tumoral heterogeneity, with the tumor center and invasive front showing identical methylation levels. Moreover, no difference in LINE-1 methylation was observed between the primary tumor and lymph node and distant metastases from the same patient. Tumor cells showed significantly less LINE-1 methylation compared to adjacent stromal and normal mucosal epithelial cells. Consequently, LINE-1 methylation was significantly lower in microdissected samples compared to macrodissected samples. A trend for less LINE-1 methylation was also observed in more advanced stages of CRC. CONCLUSIONS: LINE-1 methylation shows little intra-patient tumor heterogeneity, indicating the suitability of its use for molecular diagnosis in CRC. The methylation is relatively stable during CRC progression, leading us to propose a new concept for the association between LINE-1 methylation and disease stage.

Induction of motor neuron differentiation by transduction of Olig2 protein.

Olig2 protein, a member of the basic helix-loop-helix transcription factor family, was introduced into the mouse embryonic carcinoma cell line P19 for induction of motor neuron differentiation. We show that Olig2 protein has the ability to permeate the cell membrane without the addition of a protein transduction domain (PTD), similar to other basic helix-loop-helix transcription factors such as MyoD and NeuroD2. Motor neuron differentiation was evaluated for the elongation of neurites and the expression of choline acetyltransferase (ChAT) mRNA, a differentiation marker of motor neurons. By addition of Olig2 protein, motor neuron differentiation was induced in P19 cells.

[A case of liver metastases of sigmoid colon cancer treated effectively by oral UFT/LV].

The patient was a 77-year-old woman with multiple liver metastases of sigmoid colon cancer. She underwent low anterior resection for sigmoid colon cancer. After surgery, she selected oral administration of UFT and LV for liver metastases and multiple lymph node metastases. After two courses, the liver metastases had markedly diminished. Thirty-two months later, liver metastases had disappeared on computer tomography. This therapy was continued for five years, and recurrences are no longer shown. Severe adverse effects were not observed. Oral anti-cancer drugs can serve as effective therapy for advanced colorectal cancer of old patients.

Long interspersed nuclear element-1 hypomethylation is a potential biomarker for the prediction of response to oral fluoropyrimidines in microsatellite stable and CpG island methylator phenotype-negative colorectal cancer.

We investigated the clinical value of methylation of long interspersed nuclear element-1 (LINE-1) for the prognosis of colorectal cancer (CRC) and for the survival benefit from adjuvant chemotherapy with oral fluoropyrimidines. LINE-1 methylation in tumor DNA was measured by quantitative methylation-specific PCR in 155 samples of stage II and stage III CRC. The presence of microsatellite instability and CpG island methylator phenotype (CIMP) were assessed and 131 microsatellite stable/CIMP- cases were selected for survival analysis, of which 77 patients had received postoperative adjuvant chemotherapy with oral fluoropyrimidines. The CRC cell lines were used to investigate possible mechanistic links between LINE-1 methylation and effects of 5-fluorouracil (5-FU). High LINE-1 methylation was a marker for better prognosis in patients treated by surgery alone. Patients with low LINE-1 methylation who were treated with adjuvant chemotherapy survived longer than those treated by surgery alone, suggestive of a survival benefit from the use of oral fluoropyrimidines. In contrast, a survival benefit from chemotherapy was not observed for patients with high LINE-1 methylation. The CRC cell lines treated with 5-FU showed increased expression of LINE-1 mRNA. This was associated with upregulation of the phospho-histone H2A.X in cells with low LINE-1 methylation, but not in cells with high LINE-1 methylation. The 5-FU-mediated induction of phospho-histone H2A.X, a marker of DNA damage, was inhibited by knockdown of LINE-1. These results suggest that LINE-1 methylation is a novel predictive marker for survival benefit from adjuvant chemotherapy with oral fluoropyrimidines in CRC patients. This finding could be important for achieving personalized chemotherapy.

Repair of intra-thoracic autonomic nerves using chitosan tubes.

We investigated the safety and usefulness of chitosan nano/microfiber mesh tubes (C-tubes) in regenerating damaged thoracic autonomic nerves. Animal experiments were performed in six dogs. The right thoracic sympathetic nerve (Experiment 1, n=3) and phrenic nerve (Experiment 2, n=3) were resected and sutured with C-tubes. After surgery, in Experiment 1, Horner's syndrome was observed. In Experiment 2, mobility of diaphragm was assessed by chest X-ray imaging. Nerve regeneration was assessed pathologically in both experiments. All six dogs survived without complication throughout the observational period. In Experiment 1, sympathetic nerve began to regenerate inside the C-tube at three months. At 7 and 12 months after surgery, the sympathetic nerves were connected. Though all three dogs had right Horner's syndrome after surgery, it improved at 12 months. In Experiment 2, at 12 months, the phrenic nerves were connected in two of the three dogs. In X-ray imaging, though all three dogs had eventration of the diaphragm, the right diaphragm moved in response to breathing in the dogs in which phrenic nerve regenerated. C-tubes can be safely used to facilitate the regeneration of damaged sympathetic and phrenic nerves and the restoration of their lost functions.

A rare case of diabetic mastopathy in a Japanese man with type 2 diabetes mellitus.

We report a case of diabetic mastopathy in a man with type 2 diabetes. The patient was a 62-year-old man who had been diagnosed with type 2 diabetes at the age of 46 years. He had been treated with oral hypoglycemic agents. He noticed a mass in his left breast in February 2007, when HbA(1)c was 7.6% with the treatment using oral hypoglycemic agents, including acarbose, glimepiride, buformine, and pioglitazone. Mammography of the breast showed increased density, and ultrasonography showed a regular-shaped hypoechoic mass. Core needle biopsy was performed, and diabetic mastopathy was confirmed pathologically. Diabetic mastopathy usually occurs in women with type 1 diabetes. This case, a man with type 2 diabetes, is very rare.

[The association of expression and gene polymorphism of the thymidylate synthase gene with 5-fluoro-5'-deoxyuridine sensitivity in gastric cancer cell lines].

Thymidylate synthase (TS) is a target enzyme of 5-fluorouracil (5-FU), and the TS expression level of cancer tissues is a potential predictor of the response to 5-FU-based chemotherapy. The TS gene has a polymorphic tandem-repeat sequence, which is associated with its protein expression. Therefore, the TS polymorphism may also be a predictor of the response to 5-FU-based chemotherapy. In this study, we analyzed the TS genotype, TS protein level, and sensitivity to 5-fluoro-5'-deoxyuridine (5'-dFUrd) in 10 human gastric cancer cell lines. TS genotype was classified into 2R-homozygote (2R/2R, n=3), 3R-homozygote (3R/3R, n=5), and 2R/3R-heterozygote (2R/3R, n=2). The cell lines with 3R/3R showed a significantly higher IC50 value compared to those with 2R/2R or 2R/3R genotype. There was no relationship between TS protein level and 5'-dFUrd sensitivity. However, a statistically significant relationship was revealed between them when the subgroup with the genotypes of 2R/2R or 2R/3R was considered (r=0.815, p<0.05). In this subgroup, the cell lines with higher TS protein showed higher IC50 value for 5'-dFUrd, indicating less sensitivity to 5'-dFUrd. An identical relationship between the TS protein level and IC50 was also observed in the subgroup with 3R/3R genotype, although it did not reach statistical significance (r=0.745, p=0.09). These results suggest that the TS gene polymorphism and TS protein level may be independent predictors for 5-FU-based chemotherapy.