Oocyte-like cells (OLC) can be generated by stem cells after the induction and differentiation in vitro, and maturated when transplanted in vivo to improve the development potential. Human amniotic fluid stem cells (hAFSC) were cultured for 10 days in porcine follicle fluid (pFF) that was extracted from the medium follicle with high levels of hormones and Bmp 15 protein. After the induction, the cell aggregates showed the germ cell-like cells and produced the germ cell marker oct4, and triggered epigenetic changes with high expression of methylation transferase gene dnmt3b. The cell aggregates were packaged into porcine theca folliculi to form grafts, which were then transplanted into mouse renal capsule. After one month of transplantation, the morphology of OLC from a graft was not only similar to oocytes, but also expressed the germ cells markers (oct4, nanog, stella, ifitm3, dazl, nanos3, bmp15, and gd9). The results demonstrate that the in vivo differentiation model was useful for OLC development.
Amniotic Fluid ; cytology ; Animals ; Biomarkers ; Cell Differentiation ; Female ; Humans ; Mice ; Oocytes ; cytology ; Ovarian Follicle ; Stem Cells ; cytology ; Swine

Objective:To understand the patients'satisfaction on catastrophic medical insurance of New Rural Cooperative Medical Scheme ( NRCMS ) , and provide references for effective management and policy improvement. Methods:According to the principles of representative sampling, a questionnaire survey conducted. 484 patients got compensation for catastrophic insurance in 2013 were surveyed, and information of medical costs was analyzed. Re-sults:The average medical cost accounts for 84. 57% of total household expenditure. Mandatory medical expenditure is still high and economic burden of diseases is heavy. This policy has increased the compensation rate, and the actu-al compensation rate increases 4. 79% in three districts. The average score of patients' attitude toward catastrophic medical insurance is (3.95±1.05) points, 88. 43% of patients continue to participate in medical insurance. Con-clusions and suggestions:Patients' satisfaction on catastrophic medical insurance of NRCMS is high. It is suggested that the existing financing modes should maintain stability in the short term, and the compensatory rate should be im-proved reasonably.

Objective To review the current research and application of root cause analysis (RCA) method in China's hospital management.Methods A retrospective analysis in six aspects was made by means of formulating a retrieval strategy and inclusion criteria, retrieval of databases, and literature review.Results The number of Chinese RCA researches was increasing year by year, mostly from researcher of the eastern coastal areas.These studies focused on the effects of RCA application outcomes and nursing safety.Tools in use were mainly fishbone diagram, brainstorming and 3-why method.72.24% of the root causes as found in literature were system factors, and 27.76% were human factors.Most of the researches made positive comments on RCA.Conclusions RCA is being warmly embraced by China's hospital management as it can positively change the accountability culture towards adverse events.Yet RCA has not been satisfactorily applied, and its future research and application in China's hospital management need more in-depth study and critical analysis.

BACKGROUND:Pretreatment with moxibustion is a preventive treatment in traditional Chinese medicine.Pretreatment with moxibustion at the onset of prodromal symptoms can significantly reduce the symptoms and delay the onset of many diseases,but the exact mechanism remains to be studied. OBJECTIVE:To investigate the mechanism of SIRT1/FoxO3 pathway in moxibustion pretreatment to ameliorate oxidative stress injury in cerebral ischemia-reperfusion model rats. METHODS:Forty-eight Sprague-Dawley rats were randomly divided into sham-operated group,model group,moxibustion pretreatment group,and moxibustion pretreatment+EX527(SIRT1 inhibitor)group,with 12 rats in each group.The moxibustion pretreatment group was given moxibustion with seed-sized moxa cone at Baihui,Dazhui,and Zusanli before modeling,three moxa-cones per acupoint,once a day for 7 days.In the model group,moxibustion pretreatment group and moxibustion pretreatment+EX527 group,the rat model of middle cerebral artery occlusion was made by suturing of the middle cerebral artery 30 minutes after the last moxibustion.After 2 hours of cerebral ischemia,the middle artery suture was removed and the rats were reperfused for 12 hours.In the sham-operated group,only the common carotid artery,internal carotid artery,and external carotid artery were dissected without suturing the middle cerebral artery.In the moxibustion pretreatment+EX527 group,EX527(15 mg/kg)was given intraperitoneally 30 minutes before each moxibustion.After 12 hours of reperfusion,the rats were scored for neurological deficits,and the cerebral infarct volume was calculated by 2,3,5-triphenyltetrazolium chloride staining method.The levels of oxidative stress factors in the infarcted tissues were detected by the kit method,and western-blot method was used to detect the expression levels of SIRT1,FoxO3,p-FoxO3 and brain-derived neurotrophic factor in the ischemic area of the cerebral cortex. RESULTS AND CONCLUSION:After 12 hours of reperfusion,the neurobehavioral score in the model group was significantly higher than that in the sham-operated group(P＜0.01),while the score in the moxibustion pretreatment group was significantly lower than that in the model group(P＜0.01)and moxibustion pretreatment+EX527 group(P＜0.05).There were no obvious infarct foci in the brain tissue of the sham-operated rats,but obvious ischemic foci were observed in the right side of the brain tissue of the rats in the model group(P＜0.01).The right infarct volume in the moxibustion pretreatment group was significantly reduced compared with the model group(P＜0.01),while the right infarct volume in the moxibustion pretreatment+EX527 group was significantly enlarged compared with the moxibustion pretreatment group.After 12 hours of reperfusion,the level of malondialdehyde was significantly elevated(P＜0.01)and the expression of superoxide dismutase was significantly decreased(P＜0.01)in the model group compared with the sham-operated group.The levels of malondialdehyde was significantly decreased(P＜0.01,P＜0.05)and the expression of superoxide dismutase was significantly increased(P＜0.01,P＜0.05)in the moxibustion pretreatment group compared with the model group and the moxibustion pretreatment+EX527 group.Western blot results showed that the expression levels of SIRT1,FoxO3,p-FoxO3,and brain-derived neurotrophic factor proteins were significantly higher in the model group compared with the sham-operated group(P＜0.01);compared with the model group,the expression levels of SIRT1,FoxO3,and brain-derived neurotrophic factor were significantly higher in the moxibustion pretreatment group(P＜0.01),and p-FoxO3 expression was significantly lower(P＜0.01);compared with the moxibustion pretreatment+EX527 group,the expression levels of SIRT1,FoxO3,and brain-derived neurotrophic factor were elevated in the moxibustion pretreatment group(P＜0.05),and no statistically significant difference was found in the p-FoxO3 expression(P＞0.05).To conclude,moxibustion pretreatment can significantly improve neurological function in rats after cerebral ischemia-reperfusion,and the mechanism may be related to the activation of SIRT1/FoxO3 pathway to reduce oxidative stress injury in the rat model of cerebral ischemia-reperfusion.

BACKGROUND:It was found that moxibustion can inhibit the inflammatory factors in the serum of rats with cerebral ischemia/reperfusion injury,resist oxidative stress,inhibit cell apoptosis,and effectively reduce cerebral ischemia/reperfusion injury. OBJECTIVE:To observe the effects of different moxibustion intervention time on the expression levels of nucleotide binding oligomerization domain-like protein 3 inflammasome(NLRP3),cysteine aspartase(caspase-1),apoptosis-related speck-like protein,exfoliatin-D protein,interleukin-1β and interleukin-18 in rats with cerebral ischemia/reperfusion injury,and to explore its action mechanism. METHODS:SD rats were randomly divided into sham operation group(n=9)and operation group(n=36).The model of focal cerebral ischemia/reperfusion injury was established by middle cerebral artery occlusion in the operation group.After successful modeling,the rats in the operation group were further divided into model group,moxibustion 10-minute group,moxibustion 15-minute group and moxibustion 30-minute group,with 9 rats in each group.Rats in the moxibustion 10-minute,15-minute and 30-minute groups were given moxibustion at"Baihui,Dazhui and Zusanli",respectively,once a day for a total of 7 days.The neurological deficits of rats were evaluated by LONGA method.The cerebral infarction was observed by 2,3,5-triphenyltetrazolium chloride staining.The pathological changes of brain tissue were observed by hematoxylin-eosin staining.The contents of interleukin-1β and interleukin-18 in serum of rats in each group were detected by ELISA.Immunohistochemistry and western blot assay were used to detect the expression levels of NLRP3,caspase-1,apoptosis-related spot-like protein and gasdermin D in the ischemic cortex of rats in each group. RESULTS AND CONCLUSION:Compared with the sham operation group,the neurological deficit score of the model group was significantly increased(P<0.01).Compared with the model group,the neurological deficit score of the moxibustion groups was significantly reduced(P<0.01).Compared with the sham operation group,the infarct volume of the model group was significantly increased(P<0.01).Compared with the model group,the infarct volume of the moxibustion groups was significantly reduced(P<0.01);the infarct volume of the rats was smallest in the moxibustion 30-minute group(P<0.05).Compared with the model group,the contents of inflammatory factors interleukin-1β and interleukin-18 in the serum of rats in the moxibustion groups were decreased(P<0.01).Compared with the moxibustion 10-minute group,the contents of inflammatory factors in the serum of rats in the moxibustion 30-minute group were significantly decreased(P<0.05).Compared with the model group,the expression of NLRP3,apoptosis-related spot-like protein,Caspase-1 and gasdermin D protein in the ischemic cortex of the moxibustion groups was significantly decreased(P<0.01).Compared with the moxibustion 10-minute and 15-minute groups,the expression of protein in the moxibustion 30-minute group was significantly decreased(P<0.05).It is concluded that moxibustion at Baihui,Dazhui and Zusanli can reduce cerebral ischemia/reperfusion injury,among which moxibustion for 30 minutes has the best effect,and its mechanism may be related to the inhibition of pyroptosis mediated by NLRP3/Caspase-1 pathway.

Objective:To investigate the predictive value of peritoneal protein clearance (Pcl) for cardiovascular events and cardiovascular mortality in peritoneal dialysis (PD) patients.Methods:Eligible PD patients were prospectively enrolled from January 2014 to April 2015 in the PD Center of Renji Hospital, School of Medicine, Shanghai Jiao Tong University. All patients were followed up until death, withdrawing from PD, transferring to other centers, or the end of study period (October 1, 2018). The patients were divided into high Pcl group and low Pcl group by the median Pcl, and the differences of related indicators between the two groups were compared. A multiple linear regression model was used to analyze the influencing factors of Pcl. The Kaplan-Meier method and Log-rank test were used to compare the cumulative survival rates of patients between the two groups. A multivariate Cox regression model was used to estimate the risk of cardiovascular events and cardiovascular mortality in relation to Pcl in PD patients.Results:A total of 271 patients were enrolled, with 135 males (49.8%), age of (56.92±0.84) years old and a median PD duration of 38.77(19.00, 63.10) months. There were 70 patients (25.8%) comorbiding with diabetes and 81 patients (29.9%) with cardiovascular diseases (CVD). The median Pcl of this cohort was 67.93(52.31, 88.36) ml/d. Compared with the low Pcl group (Pcl<67.93 ml/d), the high Pcl group (Pcl≥67.93 ml/d) had older age, and greater proportion of CVD, body mass index (BMI), pulse pressure, brain natriuretic peptide, mass transfer area coefficient of creatinine (MTACcr), and lower serum albumin (all P<0.05). There was no significant difference in gender, dialysis duration, proportion of diabetes, proportion of angiotensin converting enzyme inhibitor and angiotensin receptor blocker, proportion of continuous ambulatory PD, high sensitivity C reactive protein, fluid removal including 24 h urine volume and 24 h ultrafiltration, and residual renal function between the two groups (all P>0.05). Multiple linear regression analysis showed that serum albumin ( β=-0.388, P<0.001), BMI ( β=0.189, P<0.001), and MTACcr ( β=0.247, P<0.001) were independently related to lg(Pcl). During the study period, 55 patients experienced one or more cardiovascular events and 39 patients had cardiovascular mortality. According to Kaplan-Meier analysis, cardiovascular mortality in the high Pcl group was higher than that of low Pcl group (Log-rank χ2=6.902, P=0.009). Multivariate Cox regression analysis showed that, high lg(Pcl) was an independent influencing factor of cardiovascular events in PD patients ( HR=7.654, 95% CI 1.676-34.945, P=0.009). Conclusions:Serum albumin, BMI and MTACcr are independently associated with Pcl, and Pcl is an independent predictor of cardiovascular events in PD patients.

We present GranatumX, a next-generation software environment for single-cell RNA sequencing (scRNA-seq) data analysis. GranatumX is inspired by the interactive webtool Granatum. GranatumX enables biologists to access the latest scRNA-seq bioinformatics methods in a web-based graphical environment. It also offers software developers the opportunity to rapidly promote their own tools with others in customizable pipelines. The architecture of GranatumX allows for easy inclusion of plugin modules, named Gboxes, which wrap around bioinformatics tools written in various programming languages and on various platforms. GranatumX can be run on the cloud or private servers and generate reproducible results. It is a community-engaging, flexible, and evolving software ecosystem for scRNA-seq analysis, connecting developers with bench scientists. GranatumX is freely accessible at http://garmiregroup.org/granatumx/app.