A gross anatomical study of the styloid process of the temporal bone in Japanese cadavers.

BACKGROUND: The incidence of an elongated styloid process (SP) and average length and diameter of SP have not been reported using Japanese cadavers. Data on the female-to-male ratio of an elongated SP vary. We calculated the average length and diameter of SP in Japanese cadavers and compared SP lengths between sexes. MATERIALS AND METHODS: Twenty-seven sides (right and left of bodies) in males and 51 sides in females were analysed. Measurements were obtained from the inferior external acoustic meatus to the distal tip of the SP. SP diameters at the proximal base, midpoint, and distal tip were measured. SP > 30 mm was considered elongated. We used Welch's t-test for the statistical analysis. Fisher's exact two-tailed test was also performed to analyse the female-to-male elongation ratio. A p-value < 0.05 was considered statistically significant. RESULTS: Styloid process elongation prevalence was 29.5% in our sample. The average full length was 27.04 ± 7.88 mm overall; the average diameters were 5.41 ± 1.77 mm at the proximal base and 2.21 ± 1.22 mm at the distal tip. The average SP measurement was 26.81 ± 5.92 mm in males and 27.16 ± 8.79 mm in females (p = 0.74). The female-to-male ratio of SP elongation was 1:2 (p = 0.041). Females had longer full lengths of non-elongated SPs than males (p = 0.004). Males had wider diameters at the proximal base of elongated SPs than females (p = 0.017). CONCLUSIONS: The average length of SP was 27.04 mm in the Japanese population and about 30% of the Japanese presented SP ≥ 30 mm. Male had significantly higher rate than female among the SP ≥ 30 mm, and female had significantly longer SPs than male among the SP < 30 mm. Anatomically, the SP gets narrow as distally goes. Our anatomical findings would be beneficial to creating treatment plans, diagnosis, and surgery.

Cigarette smoking and cervical cancer risk: an evaluation based on a systematic review and meta-analysis among Japanese women.

BACKGROUND: There is a body of evidence to suggest that cigarette smoking increases the risk of cervical cancer in women, but no study has examined the magnitude of the association in Japanese women. Here, we evaluated the association between cigarette smoking and the risk of cervical cancer in Japanese women based on a systematic review of epidemiological evidence. METHODS: Original data were obtained from a MEDLINE search using PubMed or from a search of the 'Ichushi' database, as well as by a manual search. Evaluation of associations was based on the strength of evidence and the magnitude of association, together with biological plausibility as evaluated previously by the International Agency for Research on Cancer. Meta-analysis of associations was also conducted to obtain a summarized overview of the data. RESULTS: We identified two cohort studies and three case-control studies. All five studies had indicated strong positive associations between cigarette smoking and the risk of cervical cancer. Our summary estimate indicated that the relative risk (RR) for individuals who had ever-smoked relative to never-smokers was 2.03 (95% confidence interval: 1.49-2.57). Four studies had also demonstrated dose-response relationships between cigarette smoking and the risk of cervical cancer. CONCLUSION: We conclude that there is convincing evidence that cigarette smoking increases the risk of cervical cancer among Japanese women.

Evidence-based cancer prevention recommendations for Japanese.

A comprehensive evidence-based cancer prevention recommendation for Japanese was developed. We evaluated the magnitude of the associations of lifestyle factors and infection with cancer through a systematic review of the literature, meta-analysis of published data, and pooled analysis of cohort studies in Japan. Then, we judged the strength of evidence based on the consistency of the associations between exposure and cancer and biological plausibility. Important factors were extracted and summarized as an evidence-based, current cancer prevention recommendation: 'Cancer Prevention Recommendation for Japanese'. The recommendation addresses six important domains related to exposure and cancer, including smoking, alcohol drinking, diet, physical activity, body weight and infection. The next step should focus on the development of effective behavior modification programs and their implementation and dissemination.

Comprehensive molecular exploration identified promoter DNA methylation of the CRBP1 gene as a determinant of radiation sensitivity in rectal cancer.

BACKGROUND: Neoadjuvant chemoradiotherapy (NCRT) for advanced rectal cancer (RC) is a well-evidenced therapy; however, some RC patients have no therapeutic response. Patient selection for NCRT so that non-responsive patients are excluded has been subjective. To date, no molecular markers indicating radiation sensitivity have been reported. METHODS: We irradiated six colorectal cancer (CRC) cell lines and identified HCT116 cells as radiation-sensitive and HCT15 and DLD-1 cells as radiation resistant. Using a microarray, we selected candidate radiation sensitivity marker genes by choosing genes whose expression was consistent with a radiation-resistant or sensitive cell phenotype. RESULTS: Among candidate genes, cellular retinol binding protein 1 (CRBP1) was of particular interest because it was not only induced in HCT116 cells by tentative 10 Gy radiation treatments, but also its expression was increased in HCT116-derived radiation-resistant cells vs parental cells. Forced expression of CRBP1 decreased the viability of both HCT15 and DLD-1 cells in response to radiation therapy. We also confirmed that CRBP1 was epigenetically silenced by hypermethylation of its promoter DNA, and that the quantitative methylation value of CRBP1 significantly correlated with histological response in RC patients with NCRT (P=0.031). CONCLUSIONS: Our study identified CRBP1 as a radiation-sensitive predictor in RC.

Do labral tears influence poor outcomes after periacetabular osteotomy for acetabular dysplasia?

AIMS: Acetabular dysplasia is frequently associated with intra-articular pathology such as labral tears, but whether labral tears should be treated at the time of periacetabular osteotomy (PAO) remains controversial. The purpose of this study was to compare the clinical outcomes and radiographic corrections of PAO for acetabular dysplasia between patients with and without labral tears pre-operatively. PATIENTS AND METHODS: We retrospectively reviewed 70 hips in 67 patients with acetabular dysplasia who underwent PAO. Of 47 hips (45 patients) with labral tears pre-operatively, 27 (25 patients) underwent PAO alone, and were classified as the labral tear alone (LT) group, and 20 (20 patients) underwent combined PAO and osteochondroplasty, and were classified as the labral tear osteochondroplasty (LTO) group. The non-labral tear (NLT) group included 23 hips in 22 patients. RESULTS: There were no significant differences between groups for post-operative Harris hip scores, degree of progression of osteoarthritis or rate of reoperation. The pre-operative alpha angle was significantly larger in the LTO group compared with the other groups (p < 0.0001). CONCLUSION: PAO provides equivalent short-term relief of pain and functional outcome in patients with or without labral tears. The rate of progression of osteoarthritis and reoperation was not significantly increased in patients with labral tears. TAKE HOME MESSAGE: PAO provides equivalent short-term pain relief and functional outcomes in patients with acetabular dysplasia with and without labral tears. We did not find significantly increased risks of progression of osteoarthritis or re-operation in those with labral tears. Cite this article: Bone Joint J 2016;98-B:741-6.

Circulating fibrocytes correlate with the asthma control test score.

BACKGROUND: Bronchial asthma is characterised by airway inflammation and remodelling with a decline of lung function. Fibrocytes are bone marrow-derived mesenchymal progenitor cells that play important roles in the pathogenesis of airway remodelling. Several clinical parameters are currently being used in routine clinical practice to assess outcome of therapy in asthma including frequency of rescue with short-acting ß2-agonist and the asthma control test. In this study, we hypothesised that asthma control test is associated with circulating levels of fibrocytes in bronchial asthma. METHODS: There were 20 patients with asthma and seven healthy controls. The number of CD45(+)Collagen I(+) circulating fibrocytes was assessed in the peripheral blood by flow cytometry. RESULTS: The number of circulating fibrocytes was significantly increased in asthma patients with moderate and severe disease compared to controls, and it was inversely correlated with % forced expiratory volume in one second and % forced vital capacity (%FVC). The frequency of inhalation of short-acting ß2 agonist and the asthma control test score was significantly and inversely correlated with the number of circulating fibrocytes. CONCLUSION: The results of this study showed that the number of circulating fibrocytes is inversely correlated with clinical asthma control parameters, further supporting the relevance of measuring circulating fibrocytes as a marker of clinical control in bronchial asthma.

Effects of IL6 C-634G polymorphism on tooth loss and their interaction with smoking habits.

OBJECTIVE: To examine the association between an IL6 (Interleukin-6) polymorphism (C-634G or rs1800796) and tooth loss, and an interaction between the polymorphism and smoking habits for the loss. MATERIAL AND METHODS: Our subjects were 4917 check-up examinees ages 35-69. They reported tooth loss and lifestyle in a questionnaire. We regressed the number of teeth on the IL6 genotype, gender, age, smoking, drinking, diabetes, hypertension, physical activity, energy intake, education, and brushing. We further estimated multivariate-adjusted odds ratios (ORs) for having <20 teeth. RESULTS: Participants with a GG genotype tended to have less teeth than those with CC; ß = -0.798 (95% confidence interval [CI] = -1.501--0.096). Subjects with a GG genotype were more likely to have <20 teeth than those with CC; OR was 1.56 (95% CI = 1.08-2.25). Association between current smoking and tooth loss was stronger among those with GG than among those with CC. In a multiple regression analysis, a significant interaction was found between GG genotype and current smoking in the prediction of tooth loss (P = 0.018). CONCLUSION: The IL6 C-634G polymorphism was significantly associated with tooth loss. Our results suggest greater effects of smoking on tooth loss in GG genotype individuals.

Rapid progressive clinical deterioration of cervical spondylotic myelopathy.

STUDY DESIGN: Retrospective clinical study. OBJECTIVE: To elucidate the pathophysiology of rapid progressive clinical deterioration following the onset of cervical myelopathy. SETTING: Spinal Injuries Center, Fukuoka, Japan. METHODS: A total of 43 cervical spondylotic myelopathy (CSM) patients were treated surgically by a senior surgeon. All patients showed intramedullary intensity changes on magnetic resonance (MR) imaging. Overall, eight patients suffered rapid progressive clinical deterioration; four of them had obvious anamnesis of minor trauma. We assessed the responsible injured segment by MR T2-weighted images. Clinical instabilities at the focal segment were evaluated using functional sagittal plain radiographs. Neurological evaluations were performed preoperatively and at 12 months postoperatively using American Spinal Injury Association (ASIA) motor scores and Japanese Orthopaedic Association (JOA) scores for cervical myelopathy. Intraoperatively, we evaluated the presence of adhesive scar tissue on the dura mater at the focal segment. RESULTS: The responsible injured segment was C3-4 in 75% of the rapid progressive (rp)-CSM and in 28.57% of the conventional CSM subjects. One with rp-CSM showed sagittal translational segmental instability. Preoperative ASIA motor scores and JOA scores in the rp-CSM were significantly lower than those in the conventional CSM subjects. Postoperative ASIA motor scores between the subjects showed no significant differences; however, postoperative JOA scores in the rp-CSM subjects were significantly lower. Moreover, an epidural membrane was observed in 62.5% of rp-CSM and 11.4% of conventional CSM subjects. CONCLUSIONS: We hypothesized that the pathophysiology of rp-CSM might be additional cervical cord disorder following the onset of cervical myelopathy. Early decompression surgery is recommended in such patients.

Cancer cell death induced by novel small molecules degrading the TACC3 protein via the ubiquitin-proteasome pathway.

The selective degradation of target proteins with small molecules is a novel approach to the treatment of various diseases, including cancer. We have developed a protein knockdown system with a series of hybrid small compounds that induce the selective degradation of target proteins via the ubiquitin-proteasome pathway. In this study, we designed and synthesized novel small molecules called SNIPER(TACC3)s, which target the spindle regulatory protein transforming acidic coiled-coil-3 (TACC3). SNIPER(TACC3)s induce poly-ubiquitylation and proteasomal degradation of TACC3 and reduce the TACC3 protein level in cells. Mechanistic analysis indicated that the ubiquitin ligase APC/C(CDH1) mediates the SNIPER(TACC3)-induced degradation of TACC3. Intriguingly, SNIPER(TACC3) selectively induced cell death in cancer cells expressing a larger amount of TACC3 protein than normal cells. These results suggest that protein knockdown of TACC3 by SNIPER(TACC3) is a potential strategy for treating cancers overexpressing the TACC3 protein.

Membrane-bound form of monocyte chemoattractant protein-1 enhances antitumor effects of suicide gene therapy in a model of hepatocellular carcinoma.

Suicide gene therapy using the herpes simplex virus thymidine kinase/ganciclovir (HSV-tk/GCV) system combined with monocyte chemoattractant protein-1 (MCP-1) provides significant antitumor efficacy. The current study was designed to evaluate the antitumor immunity of a newly developed membrane-bound form of MCP-1 (mMCP-1) in an immunocompetent mouse model of hepatocellular carcinoma (HCC). A recombinant adenovirus vector (rAd) harboring the human MCP-1 gene and the membrane-spanning domain of the CX3CL1 gene was used. Large amounts of MCP-1 protein were expressed and accumulated on the tumor cell surface. The growth of subcutaneous tumors was markedly suppressed when tumors were treated with mMCP-1, as compared with soluble MCP-1, in combination with the HSV-tk/GCV system (P<0.01). The numbers of Mac-1-, CD4- and CD8a-positive cells were significantly higher in tumor tissues (P<0.05), and tumor necrosis factor (TNF) mRNA expression levels with mMCP-1 were almost five-fold higher than those with soluble MCP-1. These results indicate that the delivery of the mMCP-1 gene greatly enhanced antitumor effects following the apoptotic stimuli by promoting the recruitment and activation of macrophages and T lymphocytes, suggesting a novel strategy of immune-based gene therapy in the treatment of patients with HCC.

Induced pluripotent stem cells generated from diabetic patients with mitochondrial DNA A3243G mutation.

AIMS/HYPOTHESIS: The aim of this study was to generate induced pluripotent stem (iPS) cells from patients with mitochondrial DNA (mtDNA) mutation. METHODS: Skin biopsies were obtained from two diabetic patients with mtDNA A3243G mutation. The fibroblasts thus obtained were infected with retroviruses encoding OCT4 (also known as POU5F1), SOX2, c-MYC (also known as MYC) and KLF4. The stem cell characteristics were investigated and the mtDNA mutation frequencies evaluated by Invader assay. RESULTS: From the two diabetic patients we isolated four and ten putative mitochondrial disease-specific iPS (Mt-iPS) clones, respectively. Mt-iPS cells were cytogenetically normal and positive for alkaline phosphatase activity, with the pluripotent stem cell markers being detectable by immunocytochemistry. The cytosine guanine dinucleotide islands in the promoter regions of OCT4 and NANOG were highly unmethylated, indicating epigenetic reprogramming to pluripotency. Mt-iPS clones were able to differentiate into derivatives of all three germ layers in vitro and in vivo. The Mt-iPS cells exhibited a bimodal degree of mutation heteroplasmy. The mutation frequencies decreased to an undetectable level in six of 14 clones, while the others showed several-fold increases in mutation frequencies (51-87%) compared with those in the original fibroblasts (18-24%). During serial cell culture passage and after differentiation, no recurrence of the mutation or no significant changes in the levels of heteroplasmy were seen. CONCLUSIONS/INTERPRETATION: iPS cells were successfully generated from patients with the mtDNA A3243G mutation. Mutation-rich, stable Mt-iPS cells may be a suitable source of cells for human mitochondrial disease modelling in vitro. Mutation-free iPS cells could provide an unlimited, disease-free supply of cells for autologous transplantation therapy.

Effects of (-)-epigallocatechin gallate on the motility and penetrability of frozen-thawed boar spermatozoa incubated in the fertilization medium.

Epigallocatechin gallate (EGCG) is the major polyphenol in green tea (Camellia sinensis) and is known for its antioxidant effects. The objective of the present study was to examine the effects of EGCG during in vitro fertilization (IVF) on the sperm quality and penetrability into oocytes. In the first experiment, the effects of concentration and incubation period of EGCG on the motility and penetrability of spermatozoa were examined. When frozen-thawed spermatozoa were incubated in IVF medium supplemented with 0 (control), 1, 50 and 100 µm EGCG for 1, 3 and 5 h, supplementation with 50 and 100 µm EGCG improved motility of the spermatozoa (p < 0.05), but not viability, as compared with the control group. When frozen-thawed spermatozoa were co-incubated with in vitro-matured (IVM) oocytes in IVF medium supplemented with 50 and 100 µm EGCG for 5 h, supplementation of EGCG had positive effects on sperm penetration rates. In the second experiment, the effects of supplementation of EGCG in IVF medium on penetrability of sperm from different boars and development of fertilized oocytes were evaluated. When frozen-thawed spermatozoa from six boars were co-incubated with IVM oocytes in IVF medium supplemented with 50 µm EGCG, the effect of EGCG on sperm penetration and development of oocytes after fertilization was found to vary with individual boar. Our results indicate that motility and penetrability of boar spermatozoa are improved by co-incubation with 50 µm EGCG, but the effects vary with individual boars.

Intensive smoking-cessation intervention in the dental setting.

Smoking exerts detrimental effects on dental treatment and oral health. Our goal was to evaluate effectiveness in terms of the abstinence rate in smoking-cessation intervention delivered by dental professionals. Individuals who were willing to quit smoking were randomly assigned to either an intervention or a non-intervention group. Intensive intervention was provided, consisting of 5 counseling sessions, including an additional nicotine replacement regimen. Reported abstinence was verified by the salivary cotinine level. Thirty-three persons in the intervention and 23 in the non-intervention group started the trial. On an intent-to-treat basis, 3-, 6- and 12-month continuous abstinence rates in the intervention group were 51.5%, 39.4%, and 36.4%, respectively, while the rates in the non-intervention group were consistent at 13.0%. Adjusted odds ratios (95% confidence interval) by logistic stepwise regression analyses were 7.1 (1.8, 28.5), 8.9 (1.7, 47.2), and 6.4 (1.3, 30.7), respectively. Intensive smoking-cessation intervention in the dental setting was therefore effective.

Periacetabular osteotomy reduces the dynamic instability of dysplastic hips.

We compared the dynamic instability of 25 dysplastic hips in 25 patients using triaxial accelerometry before and one year after periacetabular osteotomy. We also evaluated the hips clinically using the Harris hip score and assessed acetabular orientation by radiography before surgery and after one year. The mean overall magnitude of acceleration was significantly reduced from 2.30 m/s(2) (sd 0.57) before operation to 1.55 m/s(2) (sd 0.31) afterwards. The mean Harris hip score improved from 78.08 (47 to 96) to 95.36 points (88 to 100). The radiographic parameters all showed significant improvements. This study suggests that periacetabular osteotomy provides pain relief, improves acetabular cover and reduces the dynamic instability in patients with dysplastic hips.

Direct effects of protein S in ameliorating acute lung injury.

OBJECTIVE: Protein S may exert an anticoagulant activity by enhancing the anticoagulant activity of activated protein C and/or by directly inhibiting the prothrombinase complex. Protein S itself may also directly regulate inflammatory responses and apoptosis. The role of protein S in acute lung injury (ALI) was unknown. This study evaluated the effect of protein S on ALI in the mouse. METHODS: Animal ALI was induced in C57/BL6 mice by intratracheal instillation of lipopolysaccharide (LPS). Mice were treated with protein S or saline by intraperitoneal injection 1 h before LPS instillation. RESULTS: Activated protein or protein S alone and combined activated protein C + protein S therapy decreased inflammatory markers and cytokines in mice with acute lung injury. In LPS-treated mice compared with controls ALI was induced as shown by significantly increased levels of total protein, tumor necrosis factor-alpha, interleukin-6 and monocyte chemoattractant protein-1 in the bronchoalveolar lavage fluid. Mice with ALI treated with protein S had significantly decreased concentrations of tumor necrosis factor-alpha and interleukin-6 in the lung compared with untreated animals. Thrombin-antithrombin III, a marker of the activity of the coagulation cascade, was unchanged. Protein S inhibited the expression of cytokines in vitro and increased activation of the Axl tyrosine kinase pathway in A549 epithelial cells. CONCLUSION: Protein S protects against LPS-induced ALI, possibly by directly inhibiting the local expression of inflammatory cytokines without affecting coagulation.

Which is more invasive-mini versus standard incisions in total hip arthroplasty?

In this study, interleukin-6 (IL-6), C-reactive protein (CRP), and haemoglobin levels were evaluated to compare the degree of surgical invasion between mini and standard incisions in total hip arthroplasty (THA). Sixty-two patients admitted for primary cementless THA were enrolled in this randomised study. The patients were allocated to have surgery through either a mini incision of <10 cm or a standard incision of 15 cm. In each group, inflammatory responses were evaluated by IL-6, CRP, and haemoglobin levels before operation and one day after operation. Significant differences were not found in IL-6, CRP, and haemoglobin levels between both groups. At six months after surgery, there were no significant differences in postoperative Harris hip scores or radiographic evaluations between both groups. In conclusion, a 5.0 cm difference in the skin incision to the hip joint seemed to have no influence on the degree of surgical invasion during THA.

Quantitative genotyping by amplifying the polymorphic sequences of Pre-Melanosomal Protein (PMEL17) gene using real-time polymerase chain reaction in chickens.

1. This study was conducted to develop a quantitative genotyping system of chimaeric chickens by real-time PCR. 2. The polymorphisms in exons 7 and 11 of PMEL17 gene, which is one of the major genes affecting plumage colour, were identified from White Leghorn, Barred Plymouth Rock and Rhode Island Red chickens. 3. Quantitative genotyping was successfully performed by real-time PCR using polymorphic sequence-specific TaqMan Probes. 4. This methodology can support future research of germline chimaeric chickens as well as the application of germ cell transfer technique.

Structure of thymic cysts in congenital lymph nodes-lacking mice.

The ALY (aly/aly) mouse, a mutant of the C57BL/6j strain, has a severe immunodeficiency because of immature development of the immune organs. Both lymph nodes and Peyer's patches are lacking and both the thymus and spleen are small. Previous microscopical observation of their thymus glands revealed the presence of an indistinct border between the cortex and medulla, the absence of Hassal's corpuscles and the reduction of the medullary epithelial cell population. However, other microscopical findings for these glands have not yet been reported. In the present study, we performed light and electron microscopical observation of the thymus and found the consistent presence of extremely irregular shaped cystic cavities lined by microvilli-bearing epithelium in the medulla. The cysts comprised ceca and did not open into adjacent capillaries, although they contained some lymphocytes and macrophages in their lumens. In the thymus glands of normal C57BL/6j mice, only some small cysts oval in shape could be inconspicuously found in the medulla. Therefore, the thymic cysts may normally regress during thymic development, however, in ALY mice, the cysts may remain because of the organ immaturity.

Expression of androgen receptor through androgen-converting enzymes is associated with biological aggressiveness in prostate cancer.

AIMS: The association between the expression of androgen receptor (AR) or androgen-converting enzymes and malignant potential in prostate cancer (PCa) was examined. METHODS: PCa specimens from 44 cases of stage II, 10 cases of stage III, four cases of stage IV and two recurrent cases were semi-quantitatively studied with immunohistochemistry for AR and androgen-converting enzymes. RESULTS: The expression scores for AR, 5alpha-reductase type 1 (SRD5A1), 5alpha-reductase type 2 (SRD5A2), and aldo-keto reductase family 1 member C3 (AKR1C3) in the metastatic lesion of stage IV or recurrent cancer (n = 6) were 284.2 (30.1), 300 (0.0), 279.2 (51) and 254.2 (74.9), respectively; these scores were significantly higher than the respective scores of 121.8 (82.1), 135.1 (59.7), 167.0 (66.4) and 150.5 (62.8) for stage II and III cancer (n = 54) (p<0.001, p<0.001, p = 0.002 and p = 0.018, respectively). The expression scores for AR and SRD5A1 in stage II and III cancer with Gleason score 7 (n = 19) were 128.7 (72.3) and 150.5 (52.9); these were significantly higher than the scores of 78.8 (67.2) and 100.0 (39.6), respectively, for cancers with a Gleason score of < or =6 (n = 20) (p = 0.032 and p = 0.002, respectively). The expression scores for AR, SRD5A1 and AKR1C3 in stage II and III cancer with primary Gleason pattern > or =4 (n = 21) were 158.1 (84.3), 158.3 (61.1) and 173.8 (64.8); these were significantly higher than the scores of 98.6 (72.8), 120.3 (54.7) and 135.6 (57.6), respectively, for cancers with primary Gleason pattern < or =3 (n = 33) (p = 0.011, p = 0.026 and p = 0.034, respectively). Within Gleason score 9 cancer, the expression scores for AR and SRD5A1 in the primary lesion of stage IV (n = 3) were 276.7 (5.8) and 283.3 (28.9); these scores were significantly higher than the scores of 182.1 (86.0) and 140.0 (56.6), respectively, for stage II and III cancer (n = 7) (p = 0.027 and p = 0.001, respectively). CONCLUSIONS: Both AR and androgen-converting enzymes were upregulated in high-grade or advanced PCa.