Cai's prescription inhibits granulosa cell apoptosis through ARHGAP4 on poor ovarian responders.

PURPOSE: Poor ovarian response (POR) is a big challenge for in vitro fertilization. The traditional Chinese medicine, Cai's Prescription of Tonifying Kidney and Strengthening Vitals (Cai's Prescription) has yielded satisfactory results for POR treatment clinically, but systematic scientific research of Cai's Prescription is not well reported. This study aimed to investigate the clinical effect of Cai's Prescription on poor ovarian responders and its biological mechanism. METHODS: Serum was collected from poor ovarian responders, and IL-1ß, INFγ, FSH, E2 and AMH levels were analyzed by ELISA. Ovarian antral follicles were identified and counted using transvaginal ultrasound. The embryo quality grading were done on day 3 after retrieval. We used high-throughput sequencing of granulosa cells to investigate the gene transcription patterns of ovarian granulosa cells in poor ovarian responders after Cai's Prescription pretreatment. The expression level of ARHGAP4 was analyzed by quantitative real-time PCR and western blot. The effects of ARHGAP4 for granulosa cells were analyzed by CCK-8 assay, annexin-V and PI staining, ELISA and western blot. The effects of Cai's Prescription on the expression of PI3K-Akt pathway and apoptosis were analyzed by western blot. RESULTS: In this study, we found that Cai's Prescription pretreatment had the tendency to improve the ovarian reserve function and could increase the number of high quality embryos for poor ovarian responders. Through high-throughput sequencing of mRNA in granulosa cells, we discovered ARHGAP4, which is a member of GTPase-activating proteins (GAPs) may be a candidate target for POR treatment. ARHGAP4 was significantly increased in poor ovarian responders and can be recovered after Cai's Prescription pretreatment. Mechanically, combining the cell line model and clinical tissue samples, we found that ARHGAP4 can accelerate cell apoptosis and inflammation response in granulosa cells via PI3K-Akt signaling pathway. In addition, Cai's Prescription pretreatment for three months significantly reduced the high level of ARHGAP4 in poor ovarian responders. CONCLUSION: This study shows that the traditional Chinese medicine, Cai's Prescription yielded satisfactory results for poor ovarian responders clinically and ARHGAP4 may be a candidate target for POR treatment.

Analysis of Nonhomologous End Joining and Homologous Recombination Efficiency in HEK-293T Cells using GFP Based Reporter Systems.

DNA double-strand breaks (DSBs) represent the most perilous DNA lesions, capable of inducing substantial genetic information loss and cellular demise. In response, cells employ two primary mechanisms for DSB repair: nonhomologous end joining (NHEJ) and homologous recombination (HR). Quantifying the efficiency of NHEJ and HR separately is crucial for exploring the relevant mechanisms and factors associated with each. The NHEJ assay and HR assay are established methods used to measure the efficiency of their respective repair pathways. These methods rely on meticulously designed plasmids containing a disrupted green fluorescent protein (GFP) gene with recognition sites for endonuclease I-SceI, which induces DSBs. Here, we describe the extrachromosomal NHEJ assay and HR assay, which involve co-transfecting HEK-293T cells with EJ5-GFP/DR-GFP plasmids, an I-SceI expressing plasmid, and an mCherry expressing plasmid. Quantitative results of NHEJ and HR efficiency are obtained by calculating the ratio of GFP-positive cells to mCherry-positive cells, as counted by flow cytometry. In contrast to chromosomally integrated assays, these extrachromosomal assays are more suitable for conducting comparative investigations involving multiple established stable cell lines.

A study of temperature variability on admissions and deaths for cardiovascular diseases in Northwestern China.

OBJECTIVE: To explore the effect of temperature variability (TV) on admissions and deaths for cardiovascular diseases (CVDs). METHOD: The admissions data of CVDs were collected in 4 general hospitals in Jinchang City, Gansu Province from 2013 to 2016. The monitoring data of death for CVDs from 2013 to 2017 were collected through the Jinchang City Center for Disease Control and Prevention. Distributed lag nonlinear model (DLNM) was combined to analyze the effects of TV (daily temperature variability (DTV) and hourly temperature variability (HTV)) on the admissions and deaths for CVDs after adjusting confounding effects. Stratified analysis was conducted by age and gender. Then the attribution risk of TV was evaluated. RESULTS: There was a broadly linear correlation between TV and the admissions and deaths for CVDs, but only the association between TV and outpatient and emergency room (O&ER) visits for CVDs have statistically significant. DTV and HTV have similar lag effect. Every 1 ℃ increase in DTV and HTV was associated with a 3.61% (95% CI: 1.19% ~ 6.08%), 3.03% (95% CI: 0.27% ~ 5.86%) increase in O&ER visits for CVDs, respectively. There were 22.75% and 14.15% O&ER visits for CVDs can attribute to DTV and HTV exposure during 2013-2016. Males and the elderly may be more sensitive to the changes of TV. Greater effect of TV was observed in non-heating season than in heating season. CONCLUSION: TV was an independent risk factor for the increase of O&ER visits for CVDs, suggesting effective guidance such as strengthening the timely prevention for vulnerable groups before or after exposure, which has important implications for risk management of CVDs.

Ambient temperature variability and blood pressure in a prospective cohort of 50,000 Chinese adults.

Blood pressure has been shown to change by outdoor temperature, but whether intra- and inter-day temperature variability (TV) will bring higher effect on BP is not clear. Based on a prospective cohort study, the mixed effect model was selected to estimate the relationship between TV (daily temperature variability (DTV) and hourly temperature variability (HTV)) and BP (systolic blood pressure (SBP), diastolic blood pressure (DBP), pulse pressure (PP), and mean arterial pressure (MAP)) after adjusting for confounding variables. We found that there was a positive linear correlation between TV and BP. The results of DTV and HTV were basically consistent, but the effect estimates of HTV seemed to be larger. Gender, age, BMI, education level and BP status may modify the relationship between TV and BP. The effect of TV on BP was greater in non-heating season than in heating season. Our work contributes to a further macro mechanism evidence for the TV-CVDs association.

The effect and prediction of diurnal temperature range in high altitude area on outpatient and emergency room admissions for cardiovascular diseases.

PURPOSE: Diurnal temperature range (DTR) is a meteorological indicator closely associated with global climate change. Thus, we aim to explore the effects of DTR on the outpatient and emergency room (O&ER) admissions for cardiovascular diseases (CVDs), and related predictive research. METHODS: The O&ER admissions data for CVDs from three general hospitals in Jinchang of Gansu Province were collected from 2013 to 2016. A generalized additive model (GAM) with Poisson regression was employed to analyze the effect of DTR on the O&ER admissions for all cardiovascular diseases, hypertension, ischemic heart disease (IHD) and stoke. GAM was also used to preform predictive research of the effect of DTR on the O&ER admissions for CVDs. RESULTS: There were similar positive linear relationships between DTR and the O&ER visits with the four cardiovascular diseases. And the cumulative lag effects were higher than the single lag effects. A 1 °C increase in DTR corresponded to a 1.30% (0.99-1.62%) increase in O&ER admissions for all cardiovascular diseases. Males and elderly were more sensitivity to DTR. The estimates in non-heating season were higher than in heating season. The trial prediction accuracy rate of CVDs based on DTR was between 59.32 and 74.40%. CONCLUSIONS: DTR has significantly positive association with O&ER admissions for CVDs, which can be used as a prediction index of the admissions of O&ER with CVDs.

An effect of 24-hour temperature change on outpatient and emergency and inpatient visits for cardiovascular diseases in northwest China.

Some studies suggested that 24-h temperature change (TC24) was one of the potential risk factors for human health. However, evidence of the short-term effect of TC24 on outpatient and emergency department (O&ED) visits and hospitalizations for cause-specific cardiovascular diseases (CVDs) is still limited. The aim of this study is to explore the short-term effects of TC24 on O&ED visits and hospitalizations for CVDs in northwest China which is an area with large temperature variation. The O&ED visits records for CVDs of 3 general hospitals and the inpatient records for CVDs of 4 general hospitals were collected from January 1, 2013, to December 31, 2016, in Jinchang City, northwest China. Meteorological and air pollution data were also obtained during the same study period from local meteorological monitoring station and environmental monitoring station, respectively. A generalized additive model (GAM) with Poisson regression was employed to analyze the effects of TC24 on O&ED visits and hospitalizations for CVDs. V-shaped relationship were found between TC24 and O&ED visits and hospitalizations for CVDs, including total CVD, hypertension, coronary heart disease (CHD) and stroke. Stratified analysis showed that men and patients over 65 years old were more susceptible to temperature changes. The estimates in non-heating months were higher than in full year. TC24 can affect the O&ED visits and hospitalizations for CVDs in this study. This study provides useful data for policy makers to better prepare local responses to the impact of changes in temperature on population health.

Effects of cold and hot temperature on metabolic indicators in adults from a prospective cohort study.

BACKGROUND: Previous studies have found that exposed to low and high outdoor temperature was associated with cardiovascular diseases morbidity and mortality. The risk factors for cardiovascular disease include high blood lipid, high uric acid (UA) and high fasting plasma glucose (FPG). However, few studies have explored the effects of low and high temperature on these metabolic indicators. OBJECTIVE: To explore the effect of low and high temperature on metabolic indicators in adults from northwest of China. METHODS: Based on a prospective cohort study, a total of 30,759 individuals who participated in both baseline and first follow-up from 2011 to 2015 were selected in this study. The meteorological observation data and environmental monitoring data were collected in the same period. Associations between cold and hot temperature and blood lipid (total cholesterol (TC), triglycerides (TG), low density lipoprotein-cholesterol (LDL-C), and high density lipoprotein-cholesterol (HDL-C)), UA and FPG were conducted with mixed effect models after adjusting for confounding factors. RESULTS: A nonlinear relationship between outdoor temperature and metabolic indicators was found. For the cold effects, each 5 °C decrease of mean temperature was associated with an increase of 5.07% (95% CI: 3.52%, 6.63%) in TG and 2.85% (95% CI: 2.18%, 3.53%) in UA, While a decrease of 3.38% (95% CI: 2.67%, 4.09%) in HDL-C and 1.26% (95% CI: 0.48%, 2.04%) in LDL-C. For the heat effects, each 5 °C increase in mean temperature was associated with 1.82% (95% CI: 0.89%, 2.76%), 0.56% (95% CI: 0.11%, 1.00%), 5.82% (95% CI: 4.58%, 7.06%), 9.02% (95% CI: 7.17%, 10.87%), 0.20% (95% CI: 0.01%, 0.40%), and 1.22% (95% CI: 0.19%, 2.24%) decrease in TC, TG, HDL-C, LDL-C, UA and FPG. Age, smoking, drinking, high-oil diet and hyperlipidemia might modify the association between mean temperature and metabolic indicators. CONCLUSION: There was a significant effect of cold and hot temperature on metabolic indicators in a high altitude area of northwestern China. These results provide a basis for understanding the underlying mechanism of the influence of temperature on metabolic diseases.

Engineering three-dimensional labyrinthine fractal acoustic metamaterials with low-frequency multi-band sound suppression.

Acoustic metamaterials are a class of artificially periodic structures with extraordinary elastic properties that cannot be easily found in naturally occurring materials and can be applied to regulate the sound propagation behavior. The fractal configuration can be widely found in the acoustic system, like characterizing the broadband or multi-band sound propagation. This work will engineer three-dimensional (3D) labyrinthine fractal acoustic metamaterials (LFAMs) to regulate the sound propagation on subwavelength scales. The dispersion relations of LFAMs are systematically analyzed by the Bloch theory and the finite element method (FEM). The multi-bands, acoustic modes, and isotropic properties characterize their acoustic wave properties in the low-frequency regime. The effective bulk modulus and mass density of the LFAMs are numerically calculated to explain the low-frequency bandgap behaviors in specific frequencies. The transmissions and pressure field distributions of 3D LFAMs have been used to measure the ability for sound suppression. Furthermore, when considering the thermo-viscous loss on the transmission properties, the high absorptions occur within the multi-band range for low-frequency sound. Hence, this research contributes to potential applications on 3D LFAMs for multi-bands blocking and/or absorption on deep-subwavelength scales.

Association between ambient particulate matter (PM10) and incidence of diabetes in northwest of China: A prospective cohort study.

OBJECTIVES: We aimed to assess the association between long-term exposure to ambient PM10 and risk of diabetes incidence, based on the "Jinchang Cohort" platform in the Northwest of China. METHODS: We selected 19884 subjects who had not yet developed diabetes in the baseline and had completed survey information from "Jinchang Cohort". The residential address was used to match the nearest pollution monitoring station for each subject, and the average concentration of PM10 from baseline to follow-up were used as an estimate of individual exposure level. Cox regression model and restricted cubic splines functions were used to evaluate the effects of PM10 on the incidence of diabetes and the dose-response relationship after adjusting for confounding covariates. RESULTS: We observed 791 new-onset diabetics with a total follow-up of 45254.16 person-years (incidence rate of 17.48 per 1000 person-years). The risk of diabetes incidence increased by 17% (HR = 1.17, 95%CI: 1.08-1.26) per 10µg/m3 increase in environmental PM10, and the risk rises gradually with the rise of PM10 concentration. Comparing with the first quartile of PM10, the fully adjusted HRs (95%CI) for incident diabetes from the second to the fourth quartile of PM10 were 1.15 (95%CI: 0.93-1.43), 1.50 (95%CI: 1.22-1.84) and 1.44 (95%CI: 1.15-1.79), respectively (P for trend<0.001). Stratified analyses suggested that the risk of diabetes incidence associated with ambient PM10 was higher in female, young to middle-aged people, overweight and obese subjects, and subjects with FPG level at baseline lower than 5.6 mmol/L. CONCLUSIONS: Long-term exposure to ambient PM10 significantly associated with a higher risk of diabetes development. Some urgent strategies may be advocated to reduce air pollution that can aid in preventing the prevalence of diabetes in the population.

The effect of diurnal temperature range on blood pressure among 46,609 people in Northwestern China.

BACKGROUND: A large number of studies have found a positive association between diurnal temperature range (DTR) and cardiovascular diseases (CVDs) incidence and mortality. Few studies regarding the effects of DTR on blood pressure (BP) are available. OBJECTIVE: To investigate the effects of DTR on BP in Jinchang, northwestern China. METHODS: Based on a prospective cohort research, a total of 46,609 baseline survey data were collected from 2011 to 2015. The meteorological observation data and environmental monitoring data were collected in the same period. The generalized additive model (GAM) was used to estimate the relationship between DTR and BP after adjusting for confounding variables. RESULTS: Our study found that there was a positive linear correlation between DTR and systolic blood pressure (SBP) and plus pressure (PP), and a negative linear correlation between DTR and diastolic blood pressure (DBP). With a 1 °C increase of DTR, SBP and PP increased 0.058 mmHg (95%CI: 0.018-0.097) and 0.114 mmHg (95%CI: 0.059-0.168) respectively, and DBP decreased 0.039 mmHg (95%CI:-0.065 ~ -0.014). There was a significant interaction between season and DTR on SBP and PP. DTR had the greatest impact on SBP and PP in hot season. The association between DTR and BP varied significantly by education level. CONCLUSION: There was a significant association between DTR and BP in Jinchang, an area with large temperature change at high altitudes in northwestern China. These results provide new evidence that DTR is an independent risk factor for BP changes among general population. Therefore, effective control and management of BP in the face of temperature changes can help prevent CVDs.

Association between Short-Term Exposure to Air Pollution and Dyslipidemias among Type 2 Diabetic Patients in Northwest China: A Population-Based Study.

Air pollution exposure may play an adverse role in diabetes. However, little data are available directly evaluating the effects of air pollution exposure in blood lipids of which dysfunction has been linked to diabetes or its complications. We aimed to evaluate the association between air pollution and lipids level among type 2 diabetic patients in Northwest China. We performed a population-based study of 3912 type 2 diabetes patients in an ongoing cohort study in China. Both spline and multiple linear regressions analysis were used to examine the association between short-term exposure to PM10, SO2, NO2 and total cholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C). By spline analyses, we observed that the relationship between SO2 and HDL-C and LDL-C was shown to be non-linear (p_non-lin-association = 0.0162 and 0.000). An inverted U-shaped non-linear relationship between NO2 and LDL-C was found (p_non-lin-association < 0.0001). A J-shaped non-linear relationship between PM10 and TC, HDL-C (p_non-lin-association = 0.0173, 0.0367) was also revealed. In linear regression analyses, a 10 µg/m³ increment in SO2 was associated with 1.31% (95% CI: 0.40-2.12%), 3.52% (95% CI: 1.07-6.03%) and 7.53% (95% CI: 5.98-9.09%) increase in TC, TG and LDL-C, respectively. A 10 µg/m³ increment in PM10 was associated with 0.45% (95% CI: 0.08-0.82%), 0.29% (95% CI: 0.10-0.49%) and 0.83% (95% CI: 0.21-1.45%) increase in TC, HDL-C and LDL-C, respectively. For NO2, an increment of 10 µg/m³ was statistically associated with -3.55% (95% CI: -6.40-0.61%) and 39.01% (95% CI: 31.43-47.03%) increase in HDL-C and LDL-C. The adverse effects of air pollutants on lipid levels were greater in female and elder people. Further, we found SO2 and NO2 played a more evident role in lipid levels in warm season, while PM10 appeared stronger in cold season. The findings suggest that exposure to air pollution has adverse effects on lipid levels among type 2 diabetes patients, and vulnerable people may pay more attention on severe air pollution days.

Questionnaires and pocket spirometers provide an alternative approach for COPD screening in the general population.

BACKGROUND: In response to the Agency for Healthcare Research and Quality statement questioning the usefulness of "screening spirometry," the National Heart, Lung, and Blood Institute and the COPD Foundation held a consensus conference in June 2008 to establish a procedure to detect cases of COPD in the general population. Conference participants developed a three-stage approach, using a brief questionnaire, peak flow measurement with a pocket spirometer, and diagnostic quality spirometry. The overall objective of this study was to examine the usefulness of a simple questionnaire and peak flow measurement in screening for COPD in a self-selected population. We hypothesized that this combination would efficiently screen for clinically relevant COPD. METHODS: We queried individuals attending public events regarding the presence of wheeze and/or asthma, mucus production, dyspnea, exposure to irritants, and tobacco use. Peak expiratory flow (PEF) was then measured with a pocket spirometer. If PEF was < 70% predicted, spirometry was performed. In order to estimate the false-negative rate, a random sample of every 10th participant was also selected for spirometry. RESULTS: Between June 2008 and December 2009, 5,761 adults completed the risk assessment questionnaire. The mean age of the respondents was 54 years, 58% were women, and 88% were white. Of these, 5,638 participants completed pocket spirometry, and 315 (5.6%) had PEF < 70% predicted. Of 5,323 with normal PEF, 651 underwent spirometry. The performance of PEF was assessed via positive and negative predictive values relative to a diagnosis of clinically significant airflow obstruction, defined as FEV(1)/FEV(6) < the lower limit of normal and FEV(1) < 60% predicted. Of 4,238 subjects with at least two risk factors, 267 (6.3%) had PEF < 70%, compared with 48 of the 1,400 subjects (3.4%) with fewer than two risk factors (P < .001). Based on 729 participants with acceptable spirometry, 63.1% (113 of 179) of those with abnormal PEF tested positive for clinically significant airflow obstruction, compared with 5.5% (30 of 550) with normal PEF (P < .001). The estimated prevalence of significant COPD among the 5,638 screened was 8.7%, and sensitivity and specificity were 40.7% and 97.7%, respectively. CONCLUSIONS: A staged approach to COPD screening in adults is useful for detecting clinically significant airflow obstruction in our study population.

The prevalence of neck pain in migraine.

OBJECTIVE: To determine the prevalence of neck pain at the time of migraine treatment relative to the prevalence of nausea, a defining associated symptom of migraine. METHODS: This is a prospective, observational cross-sectional study of 113 migraineurs, ranging in attack frequency from episodic to chronic migraine. Subjects were examined by headache medicine specialists to confirm the diagnosis of migraine and exclude both cervicogenic headache and fibromyalgia. Details of all migraines were recorded over the course of at least 1 month and until 6 qualifying migraines had been treated. For each attack, subjects recorded the presence or absence of nausea as well as the intensity of headache and neck pain (graded as none, mild, moderate, or severe). RESULTS: Subjects recorded 2411 headache days, 786 of which were migraines. The majority of migraines were treated in the moderate pain stage. Regardless of the intensity of headache pain at time of treatment, neck pain was a more frequent accompaniment of migraine than was nausea (P< .0001). Prevalence of neck pain correlated with chronicity of headache as attacks moved from episodic to chronic daily headache. CONCLUSIONS: In this representative cross-section of migraineurs, neck pain was more commonly associated with migraine than was nausea, a defining characteristic of the disorder. Awareness of neck pain as a common associated feature of migraine may improve diagnostic accuracy and have a beneficial impact on time to treatment.