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1.  A case of a woman with late-pregnancy-onset DKA who had normal glucose tolerance in the first trimester 
Summary
Diabetic ketoacidosis (DKA) during pregnancy is a serious complication in both mother and fetus. Most incidences occur during late pregnancy in women with type 1 diabetes mellitus. We report the rare case of a woman with type 1 diabetes mellitus who had normal glucose tolerance during the first trimester but developed DKA during late pregnancy. Although she had initially tested positive for screening of gestational diabetes mellitus during the first trimester, subsequent diagnostic 75-g oral glucose tolerance tests showed normal glucose tolerance. She developed DKA with severe general fatigue in late pregnancy. The patient's general condition improved after treatment for ketoacidosis, and she vaginally delivered a healthy infant at term. The presence of DKA caused by the onset of diabetes should be considered, even if the patient shows normal glucose tolerance during the first trimester.
Learning points
The presence of DKA caused by the onset of diabetes should be considered, even if the patient shows normal glucose tolerance during the first trimester.Symptoms including severe general fatigue, nausea, and weight loss are important signs to suspect DKA. Findings such as Kussmaul breathing with ketotic odor are also typical.Urinary test, atrial gas analysis, and anion gap are important. If pH shows normal value, calculation of anion gap is important. If the value of anion gap is more than 12, a practitioner should consider the presence of metabolic acidosis.
doi:10.1530/EDM-13-0085
PMCID: PMC3975316
2.  Redistribution of resistance and sensitivity to platinum during the observation period following treatment of epithelial ovarian cancer 
Molecular and Clinical Oncology  2013;2(2):212-218.
The standard postoperative chemotherapy for epithelial ovarian cancer is a combination therapy including platinum and taxanes. The aim this study was to investigate the degree of platinum sensitivity in patients with relapsed epithelial ovarian cancer according to the treatment-free interval (TFI) and the histological tumor type. The medical records of 405 patients diagnosed with stage III/IV ovarian cancer, including 107 patients who relapsed after attaining a clinical complete response with first-line treatment, were retrospectively reviewed. The degree of platinum sensitivity was assessed by comparing the progression-free survival (PFS) following the second-line treatment. In patients with serous/endometrioid adenocarcinoma who were treated with platinum following relapse, there were significant differences in the PFS between the following groups of patients: those who relapsed within 6 months and those who relapsed between 6 and 12 months; those who relapsed between 6 and 12 months and those who relapsed between 12 and 18 months; and those who relapsed between 12 and 18 months and those who relapsed after 18 months. By contrast, in patients with clear cell/mucinous adenocarcinoma who were treated with platinum following a relapse, there were no significant differences in the PFS between patients who relapsed within 6 months and those who relapsed between 6 and 12 months, while there were significant differences in the PFS between those who relapsed between 6 and 12 months and those who relapsed after 12 months. With regard to the patients who relapsed after 12 months, the PFS of those with clear cell/mucinous adenocarcinoma was significantly shorter compared with the PFS of those with serous/endometrioid adenocarcinoma. Therefore, we considered it justified to classify patients with clear cell/mucinous adenocarcinoma who relapsed within 12 months as platinum-resistant and those who relapsed after 12 months as platinum-sensitive.
doi:10.3892/mco.2013.223
PMCID: PMC3917783  PMID: 24649335
relapsed epithelial ovarian cancer; platinum sensitivity; treatment-free interval; progression-free interval; histological type
3.  Heart Rate Variability and Hemodynamic Change in the Superior Mesenteric Artery by Acupuncture Stimulation of Lower Limb Points: A Randomized Crossover Trial 
Objective. We investigated the relationship between superior mesenteric artery blood flow volume (SMA BFV) and autonomic nerve activity in acupuncture stimulation of lower limb points through heart rate variability (HRV) evaluations. Methods. Twenty-six healthy volunteers underwent crossover applications of bilateral manual acupuncture stimulation at ST36 or LR3 or no stimulation. Heart rate, blood pressure, cardiac index, systemic vascular resistance index, SMA BFV, and HRV at rest and 30 min after the intervention were analyzed. Results. SMA BFV showed a significant increase after ST36 stimulation (0% to 14.1% ± 23.4%, P = 0.007); very low frequency (VLF), high frequency (HF), low frequency (LF), and LF/HF were significantly greater than those at rest (0% to 479.4% ± 1185.6%, P = 0.045; 0% to 78.9% ± 197.6%, P = 0.048; 0% to 123.9% ± 217.1%, P = 0.006; 0% to 71.5% ± 171.1%, P = 0.039). Changes in HF and LF also differed significantly from those resulting from LR3 stimulation (HF: 78.9% ± 197.6% versus −18.2% ± 35.8%, P = 0.015; LF: 123.9% ± 217.1% versus 10.6% ± 70.6%, P = 0.013). Conclusion. Increased vagus nerve activity after ST36 stimulation resulted in increased SMA BFV. This partly explains the mechanism of acupuncture-induced BFV changes.
doi:10.1155/2013/315982
PMCID: PMC3872304  PMID: 24381632
4.  Expression of steroid and xenobiotic receptor in uterine carcinosarcoma, leiomyosarcoma and endometrial stromal sarcoma 
Oncology Letters  2012;5(3):835-839.
We analyzed the expression of the steroid and xenobiotic receptor (SXR) in human uterine sarcomas and evaluated its clinical significance. Forty-seven cases with archival specimens were examined for SXR expression using immunohistochemistry. All cases were scored using a semi-quantitative histological scoring (HSCORE) method. Specimens with a HSCORE >40 were regarded as SXR-positive. Various clinicopathological variables, including the expression status of estrogen receptor (ER)-α, progesterone receptor (PR) and Ki67 (MIB-1) were examined. The mean SXR HSCOREs of carcinosarcoma (CS) and leiomyosarcoma (LMS) were 9.13 and 23.6, respectively, and SXR-positive rates were 3 out of 24 (12.5%) and 4 out of 17 (23.5%), respectively. SXR was not detected in endometrial stromal sarcoma (ESS). In CS cases, significant differences were detected between the expression of SXR and age and disease stages. There was no significant correlation between SXR-positive status and either disease-free survival or overall survival. Our results support an association between SXR and malignant behavior. Our results show that overexpression of SXR may represent a useful marker to identify patients with advanced-stage CS. In addition, our results showed that SXR may aid in the diagnosis of uterine sarcomas.
doi:10.3892/ol.2012.1094
PMCID: PMC3576214  PMID: 23443531
steroid and xenobiotic receptor; uterine sarcoma; immunohistochemistry; histological scoring
5.  Individual transcriptional activity of estrogen receptors in primary breast cancer and its clinical significance 
Cancer Medicine  2012;1(3):328-337.
To predict the efficacy of hormonal therapy at the individual-level, immunohistochemical methods are used to analyze expression of classical molecular biomarkers such as estrogen receptor (ER), progesterone receptor (PgR), and HER2. However, the current diagnostic standard is not perfect for the individualization of diverse cases. Therefore, establishment of more accurate diagnostics is required. Previously, we established a novel method that enables analysis of ER transcriptional activation potential in clinical specimens using an adenovirus estrogen response element–green fluorescence protein (ERE-GFP) assay system. Using this assay, we assessed the ERE transcriptional activity of 62 primary breast cancer samples. In 40% of samples, we observed that ER protein expression was not consistent with ERE activity. Comparison of ERE activity with clinicopathological information revealed that ERE activity was significantly correlated with the ER target gene, PgR, rather than ER in terms of both protein and mRNA expression. Moreover, subgrouping of Luminal A-type breast cancer samples according to ERE activity revealed that ERα mRNA expression correlated with ER target gene mRNA expression in the high-, but not the low-, ERE-activity group. On the other hand, the low-ERE-activity group showed significantly higher mRNA expression of the malignancy biomarker Ki67 in association with disease recurrence in 5% of patients. Thus, these data suggest that ER expression does not always correlate with ER transcriptional activity. Therefore, in addition to ER protein expression, determination of ERE activity as an ER functional marker will be helpful for analysis of a variety of diverse breast cancer cases and the subsequent course of treatment.
doi:10.1002/cam4.41
PMCID: PMC3544461  PMID: 23342282
Breast cancer; ERE transcriptional activity; estrogen receptor α; Ki67; Luminal A
6.  Hemodynamic Changes in the Brachial Artery Induced by Acupuncture Stimulation on the Lower Limbs: A Single-Blind Randomized Controlled Trial 
Acupuncture is commonly performed at acupoints. No comparisons of quantitative physiological alterations in the brachial artery (BA) induced by the stimulation of different acupoints in the lower limbs have been performed in humans. Therefore, we investigated changes in blood flow volume (BFV) in the BA as an indicator of the physiological effects induced by stimulation at 3 points. Seventy-five healthy participants aged 33 ± 9 years (mean ± SD) were enrolled and randomly assigned to 3 groups; they received stimulation at 3 different points located on the lower limbs: ST36, LR3, and a non-acupoint. Stimulation was performed bilaterally with manual rotation of the needles. Using ultrasonography, BFV was measured continuously from rest to 180 seconds after stimulation. LR3 stimulation significantly increased BFV compared to that before needle insertion. Meanwhile, stimulation at ST36 and the non-acupoint significantly decreased BFV compared to that before needle insertion. Stimulation at LR3 elicited a significant increase in BFV compared to that at ST36 and the non-acupoint. The results suggest that the stimulation of different points on the lower limbs causes distinct physiological effects on BFV in the BA.
doi:10.1155/2012/958145
PMCID: PMC3525317  PMID: 23304231
7.  Inflammation of the Fetal Ovine Skin Following in utero Exposure to Ureaplasma parvum 
Reproductive Sciences  2011;18(11):1128-1137.
There is increasing evidence linking in utero infection and inflammation to preterm birth. Many commensal urogenital tract microorganisms, including the Mycoplasmas and Ureaplasmas, are commonly detected in association with preterm birth. Using an ovine model of sterile fetal inflammation, we demonstrated previously that the fetal skin generates a robust inflammatory response following in utero exposure to lipopolysaccharides from Escherichia coli. The fetal skin’s response to colonization of the amniotic fluid by viable microorganisms remains unstudied. We hypothesised that in utero infection with Ureaplasma parvum serovar 3 would induce a proinflammatory response in the fetal skin. We found that (1) cultured fetal keratinocytes (the primary cellular constituent of the epidermis) respond to U. parvum exposure in vitro by increasing the expression of the chemotactant monocyte chemoattractant protein 1 (MCP-1) but not interleukin 1β (IL-1β), IL-6, IL-8, or tumor necrosis factor-α (TNF-α); (2) the fetal skin’s response to 7 days of U. parvum exposure is characterized by elevated expression of MCP-1, TNF-α, and IL-10; and (3) the magnitude of inflammatory cytokine/chemokine expression in the fetal skin is dependent on the duration of U parvum exposure. These novel findings provide further support for the role of the fetal skin in the development of fetal inflammation and the preterm birth that may follow.
doi:10.1177/1933719111408114
PMCID: PMC3343146  PMID: 22031190
preterm birth; uterine infection; inflammation
8.  Evaluation of the Effects of Acupuncture on Blood Flow in Humans with Ultrasound Color Doppler Imaging 
Color Doppler imaging (CDI) can be used to noninvasively create images of human blood vessels and quantitatively evaluate blood flow in real-time. The purpose of this study was to assess the effects of acupuncture on the blood flow of the peripheral, mesenteric, and retrobulbar arteries by CDI. Statistical significance was defined as P values less than 0.05. Blood flow in the radial and brachial arteries was significantly lower during needle stimulation on LR3 than before in healthy volunteers, but was significantly higher after needle stimulation than before. LR3 stimulation also resulted in a significant decrease in the vascular resistance of the short posterior ciliary artery and no significant change of blood flow through the superior mesenteric artery (SMA) during acupuncture. In contrast, ST36 stimulation resulted in a significant increase in blood flow through the SMA and no significant change in the vascular resistance of the retrobulbar arteries. Additionally, acupuncture at previously determined acupoints in patients with open-angle glaucoma led to a significant reduction in the vascular resistance of the central retinal artery and short posterior ciliary artery. Our results suggest that acupuncture can affect blood flow of the peripheral, mesenteric, and retrobulbar arteries, and CDI can be useful to evaluate hemodynamic changes by acupuncture.
doi:10.1155/2012/513638
PMCID: PMC3388479  PMID: 22778772
9.  Development and Clinical Application of a Precise Temperature-Control Device as an Alternate for Conventional Moxibustion Therapy 
Moxibustion therapy has been used in East Asian medicine for more than a thousand years. However, there are some problems associated with this therapy in clinical practice. These problems include lack of control over the treatment temperature, emission of smoke, and uneven temperature distribution over the treatment region. In order to resolve these problems, we developed a precise temperature-control device for use as an alternate for conventional moxibustion therapy. In this paper, we describe the treatment of a single patient with paralytic ileus that was treated with moxibustion. We also describe an evaluation of temperature distribution on the skin surface after moxibustion therapy, the development of a heat-transfer control device (HTCD), an evaluation of the HTCD, and the clinical effects of treatment using the HTCD. The HTCD we developed can heat the skin of the treatment region uniformly, and its effect may be equivalent to conventional moxibustion, without the emission of smoke and smell. This device can be used to treat ileus, abdominal pain, and coldness of abdomen in place of conventional moxibustion in modern hospitals.
doi:10.1155/2012/426829
PMCID: PMC3382393  PMID: 22754583
10.  Haemodynamic Changes in the Superior Mesenteric Artery Induced by Acupuncture Stimulation on the Lower Limbs 
Acupuncture is commonly performed on acupoints. A comparison of quantitative physiological alterations induced by stimulation on different acupoints has never been performed in the superior mesenteric artery (SMA) in humans. Therefore, we investigated changes in blood flow volume (BFV) in the SMA as an indicator of physiological effects induced by stimulation on 3 points. Thirty healthy participants aged 29 ± 10 years (mean ± SD) were enrolled. All participants underwent stimulations on 3 points located in the lower legs: ST36, LR3, and a non-acupoint. Control pertains to a condition with no-stimulation. Stimulation was performed bilaterally with manual rotation of the needles. BFV was measured by ultrasonography before insertion and 10, 20, 30, and 60 minutes after stimulation. Following acupuncture on ST36, BFV increased significantly 20 and 30 minutes after stimulation, compared to BFV before insertion (P < 0.05). Following stimulation on LR3 and the non-acupoint, no significant differences in BFV could be found. Relative to the no-stimulation group, stimulation on LR3, and the non-acupoint, stimulation on ST36 elicited a significant increase in BFV (P < 0.05). The results suggest that stimulation on the different points causes distinct physiological effects in BFV in the SMA.
doi:10.1155/2012/908546
PMCID: PMC3366235  PMID: 22675391
11.  Report on Disaster Medical Operations with Acupuncture/Massage Therapy After the Great East Japan Earthquake 
The Great East Japan Earthquake inflicted immense damage over a wide area of eastern Japan with the consequent tsunami. Department of Traditional Asian Medicine, Tohoku University, started providing medical assistance to the disaster-stricken regions mainly employing traditional Asian therapies.
We visited seven evacuation centers in Miyagi and Fukushima Prefecture and provided acupuncture/massage therapy. While massage therapy was performed manually, filiform needles and press tack needles were used to administer acupuncture. In total, 553 people were treated (mean age, 54.0 years; 206 men, 347 women). Assessment by interview showed that the most common complaint was shoulder/back stiffness. The rate of therapy satisfaction was 92.3%. Many people answered that they experienced not only physical but also psychological relief.
At the time of the disaster, acupuncture/massage therapy, which has both mental and physical soothing effects, may be a therapeutic approach that can be effectively used in combination with Western medical practices.
doi:10.4137/IMI.S9541
PMCID: PMC3342075  PMID: 22563235
Great East Japan Earthquake; Traditional Asian Medicine; acupuncture; massage; disaster
12.  High-throughput detection of aberrant imprint methylation in the ovarian cancer by the bisulphite PCR-Luminex method 
Background
Aberrant DNA methylation leads to loss of heterozygosity (LOH) or loss of imprinting (LOI) as the first hit during human carcinogenesis. Recently we developed a new high-throughput, high-resolution DNA methylation analysis method, bisulphite PCR-Luminex (BPL), using sperm DNA and demonstrated the effectiveness of this novel approach in rapidly identifying methylation errors.
Results
In the current study, we applied the BPL method to the analysis of DNA methylation for identification of prognostic panels of DNA methylation cancer biomarkers of imprinted genes. We found that the BPL method precisely quantified the methylation status of specific DNA regions in somatic cells. We found a higher frequency of LOI than LOH. LOI at IGF2, PEG1 and H19 were frequent alterations, with a tendency to show a more hypermethylated state. We detected changes in DNA methylation as an early event in ovarian cancer. The degree of LOI (LOH) was associated with altered DNA methylation at IGF2/H19 and PEG1.
Conclusions
The relative ease of BPL method provides a practical method for use within a clinical setting. We suggest that DNA methylation of H19 and PEG1 differentially methylated regions (DMRs) may provide novel biomarkers useful for screening, diagnosis and, potentially, for improving the clinical management of women with human ovarian cancer.
doi:10.1186/1755-8794-5-8
PMCID: PMC3342152  PMID: 22443985
Genomic imprinting; Ovarian cancer; DNA methylation; Bisulphite PCR-Luminex(BPL)method; LOI (loss of imprinting)
13.  Estrogen formulations and beauty care practices in Japanese women 
Purpose
Traditionally, oral estrogens have been used for hormone replacement therapy. However, in Japan, additional estrogen formulations have been used, including transdermal patches and transdermal gels. The latter have a unique commonality with cosmetics because both of them are applied to the skin. Beauty care is one of the most important lifestyle factors for women, and it has been reported that the amount of attention paid to beauty care has an effect in determining whether or not women will choose to undergo HRT during menopause. Therefore, our study focused on estrogen formulations and beauty care practices.
Patients and methods
Fifty women who use hormone replacement therapy were recruited from the outpatient clinic of Tohoku University Hospital. They were treated with oral conjugated estrogen (n = 11), transdermal 17β-estradiol patch (n = 11), and transdermal 17β-estradiol gel (n = 28). They completed a questionnaire to assess their lifestyle (beauty care practices and exercise habits) and their compliance. The transdermal gel users were further interviewed about their subjective impressions regarding “smell”, “sticky feeling”, “spreadability”, and “irritation” on the skin using a five-grade scale.
Results
There were no differences in the usability of medicines and patient compliance among the estrogen formulations. We observed a positive tendency between the level of beauty care and transdermal gel use (P = 0.0645, ordinary logistic regression analysis). The gel users placed top priority on a lack of “sticky feeling” but the subjective impression regarding “sticky feeling” was worst among the four factors (P < 0.01, Steel–Dwass test). Correspondence analysis showed that the subjective impressions of transdermal gel corresponding to usability in the range of “moderate” to “very good” and “sticky feeling” greatly affected the usability of the formulation.
Conclusion
These results suggest that the level of attention to beauty care plays some role in the choice of estrogen formulations.
doi:10.2147/IJWH.S28368
PMCID: PMC3271811  PMID: 22312196
HRT; estrogen; transdermal gel; cosmetics; subjective impression
14.  Potential role of LMP2 as tumor-suppressor defines new targets for uterine leiomyosarcoma therapy 
Scientific Reports  2011;1:180.
Although the majority of smooth muscle neoplasms found in the uterus are benign, uterine leiomyosarcoma (LMS) is extremely malignant, with high rates of recurrence and metastasis. We earlier reported that mice with a homozygous deficiency for LMP2, an interferon (IFN)-γ-inducible factor, spontaneously develop uterine LMS. The IFN-γ pathway is important for control of tumor growth and invasion and has been implicated in several cancers. In this study, experiments with human and mouse uterine tissues revealed a defective LMP2 expression in human uterine LMS that was traced to the IFN-γ pathway and the specific effect of JAK-1 somatic mutations on the LMP2 transcriptional activation. Furthermore, analysis of a human uterine LMS cell line clarified the biological significance of LMP2 in malignant myometrium transformation and cell cycle, thus implicating LMP2 as an anti-tumorigenic candidate. This role of LMP2 as a tumor suppressor may lead to new therapeutic targets in human uterine LMS.
doi:10.1038/srep00180
PMCID: PMC3240965  PMID: 22355695
15.  A novel culture system for mouse spermatid maturation which produces elongating spermatids capable of inducing calcium oscillation during fertilization and embryonic development 
Purpose
To establish an in vitro culture system for mouse round spermatids that models spermiogenesis and enables the assessment of oocyte activation ability.
Methods
Round spermatids and Sertoli cells were isolated from testicular tissues of B6D2F1 male mice and co-cultured in the presence of testosterone and recombinant FSH. Cultured spermatids were examined for morphology and condensation of nuclei, fertilization and development rate, and Ca2+ oscillation pattern after ICSI.
Results
The cultured spermatids elongated and resembled normal elongating spermatids in terms of both morphology and nuclear condensation. No significant differences in fertilization and development rates were observed between fresh and cultured elongating spermatids. Moreover, cultured spermatids showed similar Ca2+ oscillation patterns to fresh elongating spermatids during an initial stage in oocyte activation.
Conclusions
These data suggest that a co-culture system of spermatids and Sertoli cells, supplemented with testosterone and recombinant FSH, supports normal differentiation of round spermatids into elongating spermatids, as assessed by their morphology, nuclear condensation, and oocyte activation ability.
doi:10.1007/s10815-010-9442-3
PMCID: PMC2965342  PMID: 20526662
Spermatid; In vitro maturation; Co-culture; Calcium oscillation
16.  Involvement of proteasome β1i subunit, LMP2, on development of uterin leiomyosarcma 
Background:
Although the majority of smooth muscle neoplasms found in the uterus are benign, uterine leiomyosarcoma is extremely malignant, with high rates of recurrence and metastasis. The development of gynecologic tumors is often correlated with secretion of female hormone; however, the development of human uterine leiomyosarcoma is not substantially correlated with hormonal conditions, and the risk factors are unclearly understood. Importantly, a diagnostic-biomarker, which distinguishes malignant human uterine leiomyosarcoma from benign tumor leiomyoma is yet to be established.
Aims:
It is necessary to analyze risk factors associated with human uterine leiomyosarcoma, in order to establish a diagnostic-biomarker and a clinical treatment method.
Patients and Methods:
Histology and Immunofluorescence Staining: Uteri obtained from LMP2–/– mice or its parental mice (C57BL/6 mice) were fixed in 10% buffered formalin, incubated in 4% paraformaldehyde for 8 hours, and embedded in paraffin. Tissue sections (5 μm) were prepared and stained with H&E for routine histological examination or were processed further for immunofluorescence staining with appropriate antidodies. Furthermore, a total of 101 patients between 32 and 83 years of age and diagnosed as having smooth muscle tumors of the uterus were selected from pathological files. Immunohistochemistry staining for LMP2 was performed on serial human uterine leiomyosarcoma, leiomyoma and myometrium sections.
Results:
Homozygous deficient mice for a proteasome β1i subunit, LMP2 spontaneously develop uterine leiomyosarcoma, with a disease prevalence of ~40% by 14 months of age. Defective LMP2 expression in human uterine leiomyosarcoma was demonstrated, but present in human leiomyoma and myometrium.
Conclusions:
Loss in LMP2 expression may be one of the risk factors for human uterine leiomyosarcoma. LMP2 may be a potential diagnostic-biomarker and targeted-molecule for a new therapeutic approach.
doi:10.4297/najms.2011.3394
PMCID: PMC3271393  PMID: 22362447
LMP2; uterine leiomyosarcoma; uterine leiomyoma; diagnostic-biomarker
17.  Short-Term Effects of Acupuncture on Open-Angle Glaucoma in Retrobulbar Circulation: Additional Therapy to Standard Medication 
Background. The relation between glaucoma and retrobulbar circulation in the prognosis has been indicated. Purpose. To investigate the effects of acupuncture on retrobulbar circulation in open-angle glaucoma (OAG) patients. Methods. Eleven OAG patients (20 eyes with OAG) who were treated by topical antiglaucoma medications for at least 3 months were enrolled. Acupuncture was performed once at acupoints BL2, M-HN9, ST2, ST36, SP6, KI3, LR3, GB20, BL18, and BL23 bilaterally. Retrobulbar circulation was measured with color Doppler imaging, and intraocular pressure (IOP) was also measured at rest and one hour after rest or before and after acupuncture. Results. The Δ value of the resistive index in the short posterior ciliary artery (P < .01) and the Δ value of IOP (P < .01) were decreased significantly by acupuncture compared with no acupuncture treatment. Conclusions. Acupuncture can improve the retrobulbar circulation and IOP, which may indicate the efficacy of acupuncture for OAG.
doi:10.1155/2011/157090
PMCID: PMC3062145  PMID: 21437193
18.  Changes of Blood Flow Volume in the Superior Mesenteric Artery and Brachial Artery with Abdominal Thermal Stimulation 
In traditional Chinese medicine, moxibustion is a local thermal therapy that is used for several conditions. Quantifying the effects of moxibustion therapy has been difficult because the treatment temperature depends on the physician's experience, and the temperature distribution in the target area is not uniform. This prospective observational study aims to quantify the effect of local thermal stimulation to the abdomen. We developed a heat transfer control device (HTCD) for local thermal stimulation. Twenty-four healthy subjects were enrolled and they underwent abdominal thermal stimulation to the para-umbilical region with the device for 20 min. Blood flow volume in the superior mesenteric artery (SMA) and brachial artery (BA), the heart rate and the blood pressure were measured at rest, 15 min after starting thermal stimulation and 10, 20, 30 and 40 min after completing thermal stimulation. Blood flow parameters were measured by high-resolution ultrasound. In the SMA, blood flow volume was significantly increased during thermal stimulation (P < .01), as well as at 10 min (P < .01) and 20 min (P < .05) after stimulation. In the BA, blood flow volume decreased at 40 min after stimulation (P < .01). In conclusion we could quantify the effect of local thermal stimulation with an HTCD and high-resolution ultrasound. Thermal stimulation of the para-umbilical region increased blood flow in the SMA 20 min after stimulation in healthy subjects.
doi:10.1093/ecam/nep110
PMCID: PMC3095448  PMID: 19687193
19.  The shape of the sperm midpiece in intracytoplasmic morphologically selected sperm injection relates sperm centrosomal function 
Purpose
To evaluate whether the morphology of the sperm midpiece observed by high magnification microscopy relates to sperm centrosomal function.
Methods
Sperm selected by conventional microscopy were defined as controls. By high magnification microscopy, sperm with straight midpieces were defined as Group 1, while those with tapering midpieces were defined as Group 2. Heterologous ICSI of human sperm into bovine oocytes was used to assess human sperm centrosomal function and analysis of sperm aster formation.
Results
The total rate of sperm aster formation was 80.5% in Group 1, which was significantly higher (P < 0.05) than the rate of 33.3% seen for Group 2. Furthermore, sperm aster formation rates tended to be higher for Group 1 than for the controls.
Conclusions
This study demonstrates improvement of sperm aster formation rates by selecting sperm on the basis of midpiece morphology. The injection of selected sperm bearing morphologically straight midpieces may contribute to improved expression of sperm centrosomal function, providing a positive effect on fertilization after ICSI.
doi:10.1007/s10815-009-9371-1
PMCID: PMC2842867  PMID: 20012684
Intracytoplasmic morphologically selected sperm injection (IMSI); Sperm midpiece; Centrosomal function; Sperm aster formation
20.  Total Laparoscopic Conservative Surgery for an Intramural Ectopic Pregnancy 
A 38-year-old woman, gravida 3, para 1 with a history of a left salpingectomy for an ectopic pregnancy was admitted for treatment of a presumed ectopic pregnancy. Transvaginal sonography revealed an ill-defined gestational sac and fetal heart beat within the fundal myometrium adjacent to the left cornua. Laparoscopy was performed for a suspected left cornual pregnancy or intramural pregnancy. A cystic mass 3 cm in diameter was visible within the fundal myometrium. Total laparoscopic removal of the gestational sac was performed, and the uterus was preserved. Pathologic evaluation of the excised mass demonstrated chorionic villi involving the myometrium. In the literature, only one other case describing the laparoscopic removal of an intramural pregnancy has been reported. However, in the prior report, the patient still required hysterectomy after conservative surgery. Therefore, this is the first report of the successful treatment of an intramural pregnancy exclusively with laparoscopy.
doi:10.1155/2010/504062
PMCID: PMC2963121  PMID: 20981282
21.  Dietary patterns associated with fall-related fracture in elderly Japanese: a population based prospective study 
BMC Geriatrics  2010;10:31.
Background
Diet is considered an important factor for bone health, but is composed of a wide variety of foods containing complex combinations of nutrients. Therefore we investigated the relationship between dietary patterns and fall-related fractures in the elderly.
Methods
We designed a population-based prospective survey of 1178 elderly people in Japan in 2002. Dietary intake was assessed with a 75-item food frequency questionnaire (FFQ), from which dietary patterns were created by factor analysis from 27 food groups. The frequency of fall-related fracture was investigated based on insurance claim records from 2002 until 2006. The relationship between the incidence of fall-related fracture and modifiable factors, including dietary patterns, were examined. The Cox proportional hazards regression model was used to examine the relationships between dietary patterns and incidence of fall-related fracture with adjustment for age, gender, Body Mass Index (BMI) and energy intake.
Results
Among 877 participants who agreed to a 4 year follow-up, 28 suffered from a fall-related fracture. Three dietary patterns were identified: mainly vegetable, mainly meat and mainly traditional Japanese. The moderately confirmed (see statistical methods) groups with a Meat pattern showed a reduced risk of fall-related fracture (Hazard ratio = 0.36, 95% CI = 0.13 - 0.94) after adjustment for age, gender, BMI and energy intake. The Vegetable pattern showed a significant risk increase (Hazard ratio = 2.67, 95% CI = 1.03 - 6.90) after adjustment for age, gender and BMI. The Traditional Japanese pattern had no relationship to the risk of fall-related fracture.
Conclusions
The results of this study have the potential to reduce fall-related fracture risk in elderly Japanese. The results should be interpreted in light of the overall low meat intake of the Japanese population.
doi:10.1186/1471-2318-10-31
PMCID: PMC2895588  PMID: 20513246
22.  A tripartite paternally methylated region within the Gpr1-Zdbf2 imprinted domain on mouse chromosome 1 identified by meDIP-on-chip 
Nucleic Acids Research  2010;38(15):4929-4945.
The parent-of-origin specific expression of imprinted genes relies on DNA methylation of CpG-dinucleotides at differentially methylated regions (DMRs) during gametogenesis. To date, four paternally methylated DMRs have been identified in screens based on conventional approaches. These DMRs are linked to the imprinted genes H19, Gtl2 (IG-DMR), Rasgrf1 and, most recently, Zdbf2 which encodes zinc finger, DBF-type containing 2. In this study, we applied a novel methylated-DNA immunoprecipitation-on-chip (meDIP-on-chip) method to genomic DNA from mouse parthenogenetic- and androgenetic-derived stem cells and sperm and identified 458 putative DMRs. This included the majority of known DMRs. We further characterized the paternally methylated Zdbf2/ZDBF2 DMR. In mice, this extensive germ line DMR spanned 16 kb and possessed an unusual tripartite structure. Methylation was dependent on DNA methyltransferase 3a (Dnmt3a), similar to H19 DMR and IG-DMR. In both humans and mice, the adjacent gene, Gpr1/GPR1, which encodes a G-protein-coupled receptor 1 protein with transmembrane domain, was also imprinted and paternally expressed. The Gpr1-Zdbf2 domain was most similar to the Rasgrf1 domain as both DNA methylation and the actively expressed allele were in cis on the paternal chromosome. This work demonstrates the effectiveness of meDIP-on-chip as a technique for identifying DMRs.
doi:10.1093/nar/gkq200
PMCID: PMC2926594  PMID: 20385583
23.  DNA methylation errors at imprinted loci after assisted conception originate in the parental sperm 
European Journal of Human Genetics   2009;17(12):1582-1591.
There is an increased prevalence of imprinting disorders, such as Beckwith–Wiedemann syndrome, associated with human assisted reproductive technologies (ART). Work on animal models suggests that in vitro culture may be the source of these imprinting errors. However, in this study we report that, in some cases, the errors are inherited from the father. We analyzed DNA methylation at seven autosomal imprinted loci and the XIST locus in 78 paired DNA samples. In seven out of seventeen cases where there was abnormal DNA methylation in the ART sample (41%), the identical alterations were present in the parental sperm. Furthermore, we also identified DNA sequence variations in the gene encoding DNMT3L, which were associated with the abnormal paternal DNA methylation. Both the imprinting errors and the DNA sequence variants were more prevalent in patients with oligospermia. Our data suggest that the increase in the incidence of imprinting disorders in individuals born by ART may be due, in some cases, to the use of sperm with intrinsic imprinting mutations.
doi:10.1038/ejhg.2009.68
PMCID: PMC2845511  PMID: 19471309
genomic imprinting; assisted reproductive technologies; DNA methylation; oligospermic patients; DNA methyltransferase 3-like protein
24.  DNA methylation errors at imprinted loci after assisted conception originate in the parental sperm 
European Journal of Human Genetics  2009;17(12):1582-1591.
There is an increased prevalence of imprinting disorders, such as Beckwith–Wiedemann syndrome, associated with human assisted reproductive technologies (ART). Work on animal models suggests that in vitro culture may be the source of these imprinting errors. However, in this study we report that, in some cases, the errors are inherited from the father. We analyzed DNA methylation at seven autosomal imprinted loci and the XIST locus in 78 paired DNA samples. In seven out of seventeen cases where there was abnormal DNA methylation in the ART sample (41%), the identical alterations were present in the parental sperm. Furthermore, we also identified DNA sequence variations in the gene encoding DNMT3L, which were associated with the abnormal paternal DNA methylation. Both the imprinting errors and the DNA sequence variants were more prevalent in patients with oligospermia. Our data suggest that the increase in the incidence of imprinting disorders in individuals born by ART may be due, in some cases, to the use of sperm with intrinsic imprinting mutations.
doi:10.1038/ejhg.2009.68
PMCID: PMC2845511  PMID: 19471309
genomic imprinting; assisted reproductive technologies; DNA methylation; oligospermic patients; DNA methyltransferase 3-like protein
25.  Maternal Feeding Controls Fetal Biological Clock 
PLoS ONE  2008;3(7):e2601.
Background
It is widely accepted that circadian physiological rhythms of the fetus are affected by oscillators in the maternal brain that are coupled to the environmental light-dark (LD) cycle.
Methodology/Principal Findings
To study the link between fetal and maternal biological clocks, we investigated the effects of cycles of maternal food availability on the rhythms of Per1 gene expression in the fetal suprachiasmatic nucleus (SCN) and liver using a transgenic rat model whose tissues express luciferase in vitro. Although the maternal SCN remained phase-locked to the LD cycle, maternal restricted feeding phase-advanced the fetal SCN and liver by 5 and 7 hours respectively within the 22-day pregnancy.
Conclusions/Significance
Our results demonstrate that maternal feeding entrains the fetal SCN and liver independently of both the maternal SCN and the LD cycle. This indicates that maternal-feeding signals can be more influential for the fetal SCN and particular organ oscillators than hormonal signals controlled by the maternal SCN, suggesting the importance of a regular maternal feeding schedule for appropriate fetal molecular clockwork during pregnancy.
doi:10.1371/journal.pone.0002601
PMCID: PMC2432029  PMID: 18596966

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