Moxibustion alleviates decreased ovarian reserve in rats by restoring the PI3K/AKT signaling pathway / 中西医结合学报

OBJECTIVE@#Moxibustion, a common therapy in traditional Chinese medicine, has potential benefits for treating decreased ovarian reserve (DOR). The present study investigates the protective effect of moxibustion in a rat model of DOR and explores the possible mechanisms.@*METHODS@#Sixty-four female Sprague-Dawley rats were randomly divided into four groups: control, DOR, moxibustion (MOX), and hormone replacement therapy (HRT). The DOR rat model was established by intragastric administration of 50 mg/kg Tripterygium glycoside suspension (TGS), once daily for 14 days. MOX and HRT treatments were given from the day TGS administration was initiated. The ovarian reserve function was evaluated by monitoring the estrus cycle, morphological changes in ovaries, levels of serum estradiol (E2), follicle-stimulating hormone (FSH), luteinizing hormone (LH), and anti-Mullerian hormone (AMH), pregnancy rate and embryo numbers. Terminal-deoxynucleotidyl transferase-mediated nick-end-labeling staining was used to identify ovarian granulosa cell apoptosis, while the protein and mRNA expressions of Bax, B-cell lymphoma-2 (Bcl-2), phosphatidylinositol 3-kinase (PI3K) and protein kinase B (AKT) in ovarian tissues were examined by immunohistochemistry, Western blot and quantitative reverse transcription-polymerase chain reaction.@*RESULTS@#Compared with the DOR group, MOX improved the disordered estrous cycle, promoted follicular growth, reduced the number of atresia follicles, increased the concentrations of serum E2 and AMH, and decreased serum FSH and LH concentrations. More importantly, the pregnancy rate and embryo numbers in DOR rats were both upregulated in the MOX treatment group, compared to the untreated DOR model. Further, we found that the MOX group had reduced apoptosis of ovarian granulosa cells, increased Bcl-2 expression and reduced expression of Bax. Furthermore, the PI3K/AKT signaling pathway was triggered by the moxibustion treatment.@*CONCLUSION@#Moxibustion improved ovarian function and suppressed apoptosis of ovarian granulosa cells in a rat model of DOR induced by TGS, and the mechanism may involve the PI3K/AKT signaling pathway.

Effect of acupuncture on oxidative stress and apoptosis in ovarian hypofunction induced by Tripterygium Glycosides / 针刺研究

OBJECTIVE: To explore whether manual acupuncture stimulation of "Shenshu " (BL23), "Guanyuan" (CV4), "Zhongwan" (CV12) can improve the ovarian function by resisting oxidant stress and reducing apoptosis of granulosa cells in rats with ovarian hypofunction. METHODS: Forty female SD rats with normal estrous cycles were randomly divided into blank control, model, hormone therapy and acupuncture groups (n=10 rats in each group). The ovarian hypofunction model was established by gavage of Tripterygium Glycosides suspension (50 mg·kg-1·d-1) for 14 successive days. Rats of the hormone therapy group were treated by gavage of estrogen and progesterone, and those of the acupuncture group treated by manual acupuncture stimulation of bilateral BL23 or CV4 and CV12 alternatively by using uniform reinforcing-reducing method for 10 s every 5 min (3 times in 10 min). The treatment was performed once daily for 14 days. The blank group was given equal volume of normal saline daily. On the 9th day, the estrous cycle of each rat was observed by means of vaginal smear test. The ovarian index, serum estradiol (E2), follicle stimulating hormone (FSH), luteinizing hormone (LH), malondialdehyde (MDA) and superoxide dismutase (SOD) contents were detected by ELISA. Histopathological changes of the ovary tissue were observed by H.E. staining. The expressions of apoptosis-related proteins Bax and Bcl-2 and their mRNAs of the ovaries were determined by immunohistochemical staining and quantitative real time-PCR separately. RESULTS: The menstrual disorder rate was 100% in the model group, and was significantly higher than those in the hormone therapy (30%) and acupuncture (40%) groups (P<0.01). Following modeling, the ovarian wet weight and index, E2 and SOD contents, Bcl-2 protein and mRNA expressions were significantly decreased (P<0.01, P<0.05), and serum FSH, LH and MDA levels, Bax protein and mRNA expressions were significantly increased (P<0.05, P<0.01) in the model group in comparison with the blank control group. H.E. staining showed a large amount of connective tissue in the ovary, with fewer mature follicles, increase of atresia follicles, significant reduction of luteal tissue, and appearance of scarring tissue in the model group. Compared with the model group, there were more mature follicles, fewer atresia follicles, lower abnormal granulosa cell morphology and lower estrous cycle disorder ratio in both hormone therapy and acupuncture groups. After the interventions, the decreased ovarian wet weight and index, serum E2 and SOD contents, expressions of Bcl-2 protein and mRNA were significantly increased (P<0.05, P<0.01), and the increased levels of FSH and LH, MDA, expressions of Bax protein and mRNA were significantly decreased in both hormone therapy and acupuncture groups (P<0.05, P<0.01). The effect of acupuncture was significantly superior to that of hormone in up-regulating SOD, Bax and Bcl-2 protein expressions (P<0.01). CONCLUSION: Acupuncture can improve the menstrual disorder in rats with ovarian hypofunction, which is closely related to its effects in improving antioxidant stress ability and regulating the expressions of apoptosis-related protein and mRNA of ovarian gra-nulosa cells.

Increased cancer incidence of Changhua residents living in Taisi Village north to the No. 6 Naphtha Cracking Complex.

BACKGROUND/PURPOSE: Cancer risks of residents living north to the No. 6 Naphtha Cracking Complex has not been studied before. METHODS: Our study subjects were recruited in 2014-16 from three zones north to the No. 6 Naphtha Cracking Complex, which included 229 participants from the Taisi Village (average 5.5 km from the complex), 1333 participants from the other 14 villages in Dacheng Township (9.2 km), and 372 participants from the Zhutang Township (19.9 km). Their occurrence of cancer in years-post-complex-operation (YPO) was defined by having a new cancer recorded (ICD-9: 140-208) in National Health Insurance Research Database since 1999. Poisson regression was conducted to compare incidence rate ratio among three zones in 10-16 YPO. RESULTS: We found that all-cause cancer incidence of 10-16 YPO (per 1,000 person-years) in Taisi Village (8.44) was higher than that in Dacheng (3.42) and Zhutang (2.72). Taisi residents had significantly higher concentrations of V, Cr, Mn, Ni, Cu, As, Cd, and Tl than Dacheng and Zhutang residents. The all-cause cancer incidence rate ratio between 10-16 and 0-9 YPO was 8.44 for Taisi residents. All-cause cancer incidence rate of Taisi residents was 2.55 times higher than Dacheng residents (95% CI: 1.89-3.45) and 2.43 times higher than Zhutang residents (95% CI: 1.54-3.84) in 10-16 YPO. CONCLUSION: We conclude that all-cause cancer risk was significantly increased for Taisi residents living near the No. 6 Naphtha Cracking Complex for 10-16 years after the complex began operating.

Increased cancers among residents living in the neighborhood of a petrochemical complex: A 12-year retrospective cohort study.

This study investigates whether cancers are increased for residents living in the vicinity of a petrochemical complex with coal power plants and refineries. We recruited a residential cohort of 2388 long-term residents aged above 35 years in 2009-2012 who lived within a 40 km radius of the complex. We measured their internal exposure biomarkers of urinary carcinogenic metals and retrospectively compared cancer incidences between those who lived fewer than 10 km from the complex (high exposure, HE) and those who lived more than 10 km from the complex (low exposure, LE). Residents had lived in their respective areas for 12 years, since the complex began operating in mid-1999. This included two periods of operation: 0-9 years and 10-12 years. Crude cumulative incident rates (CIRs) of all cancers were calculated for new cancer cases (ICD-9: 140-165, 170-176, 179-208) recorded in the Taiwan Health Insurance Database over total person-years at risk in each study period. Poisson regression was applied to estimate relative risks for the CIRs of all cancers between HE and LE areas during the 10-12 years since the beginning of the complex's operation, adjusting for age, gender, body mass index, smoking, hepatitis C, and occupational exposure. We found that our study subjects in HE areas had higher urinary carcinogenic metal levels, including As, Cd, Hg, Pb, and V, and higher prevalence rates of hepatitis C than those in LE areas. After the complex had been operating for 10-12 years, SIRs per 1000 person-years for all cancers in HE and LE areas were 4.44 vs. 2.48 for all subjects, 15.2 vs. 4.86 for elder subjects aged above 60 years, and 2.94 vs. 2.71 for female subjects. Correspondingly, the adjusted relative risks of CIRs for all cancers between HE and LE areas were 1.29 (95% CI: 0.99-1.68) for all subjects, 1.52 (1.04-2.22) for elder subjects, 1.41 (1.00-1.97) for female subjects, and 1.91 (1.15-3.19) for female elderly subjects. We conclude that elder and female residents living within 10 km of a petrochemical complex had higher carcinogenic exposure and cancers than those living farther away from the complex after the complex had been operating for 10 years.

Increased incidence of allergic rhinitis, bronchitis and asthma, in children living near a petrochemical complex with SO2 pollution.

This study aims to investigate incidence of allergic rhinitis, bronchitis and asthma, in children living near a petrochemical complex with SO2 pollution obtained by air monitoring stations. A total of 587 children aged 11 to 14 were recruited and classified into high and low exposure groups based on a radius of 10km from the complex. To study the influence of health on children since the operation of complex in 1999 and observe the difference of these diseases' short-term and long-term impact, we obtained the incidence rates of allergic rhinitis (ICD-9: 477), bronchitis (490-491) and asthma (493) from the Taiwan Health Insurance Database for three periods: 1999-2002, 1999-2006, and 1999-2010. Since 2001, the mean and 99th percentile of SO2 concentrations in the high exposure area have been significantly higher than those in low exposure area. There were significant differences between the high and low exposure groups in the percentage of smoking, alcohol consumption, passive smoking exposure and incense burning habits. The incidence rates of three intervals were 26.9%, 35.7%, 41.7%; 8.3%, 8.8%, 10.2%; 18.5%, 25.0%, 26.9% for allergic rhinitis, bronchitis and asthma in high exposure group. Significant differences were found between groups for allergic rhinitis in all periods, bronchitis in the first two periods, and asthma in the first period using Student's t-test. After we adjusted age, gender, group, living near roads, incense burning and passive smoking exposure, the hazard ratios between exposure groups were 3.05, 2.74, and 1.93 for allergic rhinitis with significant difference in three periods, and 2.53, 1.92 and 1.72 for bronchitis with significant difference in first period and 1.60, 1.28 and 1.29 for asthma with significant difference in first period by Cox regression. The higher incidence of allergic rhinitis was related to boys and living near roads and the higher incidence of asthma was also related to younger children, boys, and passive smoking exposure.

Statistical inference on censored data for targeted clinical trials under enrichment design.

For the traditional clinical trials, inclusion and exclusion criteria are usually based on some clinical endpoints; the genetic or genomic variability of the trial participants are not totally utilized in the criteria. After completion of the human genome project, the disease targets at the molecular level can be identified and can be utilized for the treatment of diseases. However, the accuracy of diagnostic devices for identification of such molecular targets is usually not perfect. Some of the patients enrolled in targeted clinical trials with a positive result for the molecular target might not have the specific molecular targets. As a result, the treatment effect may be underestimated in the patient population truly with the molecular target. To resolve this issue, under the exponential distribution, we develop inferential procedures for the treatment effects of the targeted drug based on the censored endpoints in the patients truly with the molecular targets. Under an enrichment design, we propose using the expectation-maximization algorithm in conjunction with the bootstrap technique to incorporate the inaccuracy of the diagnostic device for detection of the molecular targets on the inference of the treatment effects. A simulation study was conducted to empirically investigate the performance of the proposed methods. Simulation results demonstrate that under the exponential distribution, the proposed estimator is nearly unbiased with adequate precision, and the confidence interval can provide adequate coverage probability. In addition, the proposed testing procedure can adequately control the size with sufficient power. On the other hand, when the proportional hazard assumption is violated, additional simulation studies show that the type I error rate is not controlled at the nominal level and is an increasing function of the positive predictive value. A numerical example illustrates the proposed procedures.

An inferential procedure for the probability of passing the USP dissolution test.

Dissolution is one of the tests that is required and specified by the United States Pharmacopeia and National Formulary (USP/NF) to ensure that the drug products meet the standards of the identity, strength, quality, purity, and stability. The sponsors also establish the in-house specifications for the mean and standard deviation of the dissolution rates to guarantee a high probability of passing the USP/NF dissolution test. However, the USP/NF dissolution test is a complicated three-stage sampling plan that involves both the sample mean dissolution rate of all units and the dissolution rate of individual units. It turns out that the true probability of passing the USP/NF dissolution is formidable to compute analytically even when the population mean and variance of dissolution rates are known. It is not clear that previously proposed methods are the estimators of the true probability for passing the USP dissolution test. Therefore, we propose to employ a parametric bootstrap method in conjunction with the Monte Carlo simulation to obtain the sampling distribution of the estimated probabilities of passing the USP/NF dissolution test and hence the confidence interval for the passing probability. In addition, a procedure is proposed to test whether the true probability of passing the USP/NF dissolution test is greater than some specified value. A numerical example illustrates the proposed method. Copyright © 2011 John Wiley & Sons, Ltd.

Elevated blood pressure, decreased heart rate variability and incomplete blood pressure recovery after a 12-hour night shift work.

Shift work has been associated with increased risk of cardiovascular disease. This study was designed to determine the hemodynamic effects of 12-hour (12-h) shifts, and changes in blood pressure (BP) and heart rate variability (HRV) during 36 h rest time following 12-h shifts. Fifteen male shift workers with a mean age of 32.9 yr were recruited from a semiconductor factory. Ambulatory BP (AmBP) monitoring was performed for a total of 48 h for each participant. Six workers were monitored for 48 h by Holter electrocardiogram on both the day and night shifts. Paired self-comparison was used to estimate the difference between two hourly measurements of 12-h BP, HR, and HRV using the same timetable intra-individually. We also applied mixed models to estimate the effects of 12-h shifts on the delayed recovery of BP and heart rate (HR) in six workers who completed 96-h AmBP monitoring, including a 48-h night shift-rest period and another day shift period. Results showed that 12-h night shift work gave a persistently elevated systolic and diastolic BP (SBP and DBP) and HR, and decreased HRV compared to 12-h day shift work with the corresponding resting time. In addition, there was delayed SBP and DBP recovery on the first 12-h rest time in night shift workers, which was further demonstrated on the second 12-h rest time after adjustment for possible confounders through mixed models. In conclusion, 12-h night shift work may elevate BP and HR and decrease HRV. It is also associated with delayed BP recovery.

Health-related quality of life after laparoscopically assisted vaginal hysterectomy: is uterine weight a major factor?

OBJECTIVE: To assess uterine size, symptom severity, and hemoglobin level as determinants of health-related quality of life (HRQOL) in women subsequently undergoing laparoscopically assisted vaginal hysterectomy (LAVH). METHODS: Sixty-one consecutive women with uterine leiomyoma or adenomyosis undergoing LAVH were studied using a prospective cohort design. The Chinese version of the Uterine Fibroid Symptom and Quality of Life (UFS-QOL) questionnaire was assessed preoperatively. The Taiwan brief version of the World Health Organization Quality of Life (WHOQOL-BREF) questionnaire and a self-assessment of the perceived health status were assessed preoperatively and 1 day, 1 week, 12 weeks, and 18 months postoperatively. RESULTS: Women with a greater uterine weight did not report a greater severity of symptoms than those with lower uterine weight. Women with more severe symptoms had lower preoperative hemoglobin levels and were usually younger. Their perceived health status and their scores in physical domain of WHOQOL-BREF were also significantly lower, indicating a poorer HRQOL. The mixed-effects model found that a normal (higher) baseline hemoglobin level and a greater symptom severity were associated with a significant improvement of HRQOL in the physical domain of WHOQOL-BREF following LAVH. CONCLUSIONS: Preoperative symptom severity, but not uterine weight, was associated with long-term improvement in HRQOL after LAVH. Women with severe symptoms could be considered for LAVH before development of anemia.

Twenty-four-hour ambulatory blood pressure and duration of hypertension as major determinants for intima-media thickness and atherosclerosis of carotid arteries.

The relationship between time factors of elevated blood pressure (BP) and carotid atherosclerosis (CA) is still unclear. The associations between time-weighted average 24 h ambulatory systolic BP (TWA-SBP), duration of hypertension in years (hypertension-year), and CA were investigated in a petrochemical company sample of 95 executives and 91 gender- and age-matched non-executives employees. Intima-media thickness (IMT) and plaque scores of extracranial carotid artery (ECCA) were determined bilaterally by high-resolution B-mode ultrasound. The determinants of segment-specific carotid IMT and odds ratios for CA, in terms of thicker IMT (IMT > or = 75th percentile) and ECCA score > or = 3, were evaluated by multivariate regression analysis. Results revealed TWA-SBP and hypertension-year were two major determinants of IMT at common carotid artery (CCA) and carotid bulb by using mixed regression models. However, TWA-DBP was a negative determinant of IMT at CCA and carotid bulb. Meanwhile, the executives were found to be a negative association with IMT at carotid bulb. Measurements at both internal carotid artery and bulb identified duration of diabetes mellitus as significant determinant of IMT. After controlling covariates, multivariate logistic regression analysis identified TWA-SBP and hypertension-year as the important determinants for thicker IMT and ECCA > or = 3. And, TWA-DBP was found as a negative determinant for CA. In conclusion, both TWA-SBP and hypertension-year were two major determinants for carotid IMT and CA, which seem to imply that both short-term and long-term durations of elevated BP are probably crucial in the pathogenesis of CA.

Homocysteine, circulating vascular cell adhesion molecule and carotid atherosclerosis in postmenopausal vegetarian women and omnivores.

Since the adoption of vegetarian diets as a healthy lifestyle has become popular, the cardiovascular effects of long-term vegetarianism need to be explored. The present study aimed to compare the presence and severity of carotid atherosclerosis (CA), and the blood levels of Vitamin B12, homocysteine (Hcy) and soluble vascular cell adhesion molecule-1 (sVCAM-1) between 57 healthy postmenopausal vegetarians and 61 age-matched omnivores. Carotid atherosclerosis, as measured by ultrasound, was found to be of no significant difference between the two groups. Yet, fasting blood glucose, low-density lipoprotein cholesterol, and Vitamin B12 were significantly lower, while Hcy and sVCAM-1 were higher in the vegetarians as comparing with the omnivores. Multivariate regression analysis showed that the level of Vitamin B12 was negatively associated with the level of Hcy. Vegetarianism itself and Hcy level were significantly associated with sVCAM-1 level in univariate analysis; however, after adjustment for covariates, we identified age but not vegetarianism as the determinant of sVCAM-1 level. Multiple linear regression analysis identified age and systolic blood pressure, but not vegetarianism, as determinants of common carotid artery IMT. In conclusion, there was no significant difference in CA between apparently healthy postmenopausal vegetarians and omnivores. The findings of elevated Hcy in vegetarians indicate the importance of prevention of Vitamin B12 deficiency.