ErZhiTianGui Decoction alleviates age-related ovarian aging by regulating mitochondrial homeostasis and inhibiting ferroptosis.

AIM: This study was designed to investigate the pharmacological effects and mechanisms of ErZhiTianGui Decoction (EZTG) for age-related ovarian aging in mice. METHODS: This study used naturally aging mice as a model, and EZTG was used for intragastric administration. Ovarian pathological changes, as well as follicular reserve were assessed by hematoxylin and eosin staining, and serum hormone levels (anti-mullerian hormone, follicle-stimulating hormone), mitochondrial reactive oxygen species (ROS) and mitochondrial DNA (mtDNA) damage marker 8-hydroxy-2'-deoxyguanosine(8-OHdG), and lipid peroxidation markers glutathione(GSH) and malondialdehyde(MDA) were determined by enzyme-linked immunosorbent assay. Mitochondrial membrane potential (MMP) levels in ovaries were determined using flow cytometry. The levels of PINK1 and Parkin were observed using immunofluorescence staining. Mitochondrial-derived vesicles (MDVs) and mitochondrial morphology were observed using electron microscopy. Prussian blue staining was used to observe iron ion aggregation in ovarian tissue. The Iron assay kits detected total iron levels. Western blot was used to detect the expression of proteins related to mitochondrial and ferroptosis related genes. RESULTS: After EZTG treatment, aged mice showed increased ovarian reserve, improved serum hormone levels, increased MMP, GSH levels, and decreased mitochondrial ROS, 8-OHdG, and MDA levels. Immunofluorescence staining showed decreased levels of PINK1 and Parkin, and the same trend was observed for the Western blot. Meanwhile, electron microscopy showed that EZTG improved the mitochondrial morphology in the ovaries of aged mice. EZTG also decreased the total iron and protein levels of Acyl-CoA synthetase long-chain family4 (ACSL4) and increased the protein level of glutathione peroxidase 4 (GPX4) in the ovaries of aged mice. CONCLUSIONS: EZTG can maintain PINK1/Parkin-mediated mitochondrial homeostasis, reduce the lipid peroxidation caused by the accumulation of ROS, and inhibit the occurrence of ferroptosis and delaying ovarian aging. These findings suggest that EZTG may be a promising drug for treating age-related ovarian aging in females.

Mitochondrial Function Reveals Connotation of "Seven-Seven" Theory of Huangdi's Internal Classic and Mechanism of Herbal Intervention for Elderly Women Undergoing IVF-ET / 中国实验方剂学杂志

ObjectiveTo study the changes of mitochondrial function of ovarian granulosa cells in women of different ages and the effect of Erzhi-Tiangui prescription on in vitro fertilization-embryo transfer （IVF-ET） outcomes for elderly women， so as to verify the connotation of the "Seven-Seven" theory in the Huangdi's Internal Classic （《黄帝内经》）. MethodA total of 150 infertility patients undergoing IVF-ET at the Reproductive and Genetic Center of Integrative Medicine， Affiliated Hospital of Shandong University of Traditional Chinese Medicine were recruited and assigned into "hree-Seven/Four-Seven （30 cases）， Five-Seven （60 cases）， and Six-Seven （60 cases） groups according to the "Seven-Seven" theory. The Five-Seven and Six-Seven groups were further assigned into control and Chinese medicine subgroups using the random number plus envelope method， and the Chinese medicine group was administrated with Erzhi Tiangui prescription from the start day of controlled ovulation stimulation cycle to the trigger day. The IVF outcome was observed， and Western blot was employed to determine the levels of mitofusin 1 （MFN1）， mitofusin 2 （MFN2）， and dynamin-related protein 1 （Drp1） in the ovarian granulosa cells. ResultCompared with the Three-Seven/Four-Seven group， the control subgroups of the Five-Seven and Six-Seven groups showed decreased retrieved oocytes， two pronuclear （2PN） embryos， available embryos， high-quality embryos， clinical pregnancy rate， and live birth rate （P<0.05）. Moreover， the control subgroup of the Six-Seven group showed decreased fresh embryo transfer rate（P<0.05）. Compared with the control subgroup of the Five-Seven group， that of the Six-Seven group showed reduced retrieved oocytes， 2PN embryos， available embryos， high-quality embryos， and clinical pregnancy rate （P<0.05）. The Chinese medicine subgroup had more retrieved oocytes， 2PN oocytes， and available embryos than the control subgroup in the Five-Seven groups （P<0.05）. The Chinese medicine subgroup had more retrieved oocytes， than the control subgroup in the Six-Seven groups （P<0.05）. The control subgroup of the Six-Seven group showed lower expression levels of Mfn1 and Mfn2 and higher level of Drp1 than the control subgroup of the Five-Seven group （P<0.05）， which indicated that the levels of Mfn1 and Mfn2 in ovarian granulosa cells were down-regulated while the expression of Drp1 was up-regulated with aging （P<0.05）. The Chinese medicine subgroup had higher Mfn2 level and lower Drp1 level than the control subgroup in the Five-Seven group （P<0.05）， and the Chinese medicine subgroup had higher Mfn1 and Mfn2 levels and lower Drp1 level than then control subgroup in the Six-Seven group （P<0.05）. ConclusionsThe prognosis of IVF in women after "Five-Seven" became worse with aging， and the mitochondria in ovarian granulosa cells showed decreased fusion ability and increased fission， which verified the connotation of the "Seven-Seven" theory from the mitochondrial function. Erzhi Tiangui prescription can regulate the mitochondrial function of ovarian granulosa cells in elderly women， up-regulate the expression levels of Mfn1 and Mfn2 to promote mitochondrial fusion， and down-regulate the expression of Drp1 to reduce mitochondrial fission， thus alleviating the ovarian hypofunction caused by aging， improve the development potential of oocytes， and improve the IVF outcomes of elderly women. However， this prescription has limited efficacy for the elderly women in the age range of "Six-Seven".

Prevention and Treatment of Age-related Ovarian Hypofunction with Traditional Chinese Medicine： A Review / 中国实验方剂学杂志

Age-related ovarian hypofunction includes a decrease in follicle quantity and quality as well as alterations in the ovarian microenvironment，the mechanisms of which are mainly related to mitochondrial dysfunction，free radical and antioxidant systems，telomere and telomerase alterations，and apoptosis，and is one of the major factors contributing to infertility in advanced maternal age （AMA）. Despite the tremendous progress in assisted reproductive technology in recent decades，few breakthroughs have been made in alleviating age-related ovarian hypofunction and improving reproductive outcomes for AMA. In recent years，there has been an increasing number of studies on the multi-level and multi-targeted mechanisms of traditional Chinese medicine （TCM） to improve age-related ovarian hypofunction by modulating mitochondrial homeostasis，alleviating oxidative stress，and inhibiting apoptosis，while more high-quality randomized controlled trials have demonstrated the clinical efficacy of TCM in assisted reproductive technology. Given this，this article presented a systematic review of recent research and randomized controlled trials on the mechanism of Chinese medicine active ingredients，single Chinese medicine， and Chinese medicine compounds in delaying age-related ovarian hypofunction，to clarify the current status and shortcomings of the research. This paper provides medication management of TCM for effectively alleviating age-related ovarian hypofunction and improving reproductive outcomes for AMA.

Numerical assessment of impeller features of centrifugal blood pump based on fast hemolysis approximation model / 生物医学工程学杂志

The impeller profile, which is one of the most important factors, determines the creation of shear stress which leads to blood hemolysis in the internal flow of centrifugal blood pump. The investigation of the internal flow field in centrifugal blood pump and the estimation of the hemolysis within different impeller profiles will provide information to improve the performance of centrifugal blood pump. The SST kappa-omega with low Reynolds correction was used in our laboratory to study the internal flow fields for four kinds of impellers of centrifugal blood pump. The flow fields included distributions of pressure field, velocity field and shear stress field. In addition, a fast numerical hemolysis approximation was adopted to calculate the normalized index of hemolysis (NIH). The results indicated that the pressure field distribution in all kinds of blood pump were reasonable, but for the log spiral impeller pump, the vortex and backflow were much lower than those of the other pumps, and the high shear stress zone was just about 0.004%, and the NIH was 0.0089.

Relation and prevention between aspirin and relapsing haemorrhage after operation in cerebral haemorrhage patients / 中华急诊医学杂志

Objective To explore the relation and measures prevention between aspirin and relapsing haemorrhage after operation in cerebral haemorrhage patients. Method It' s a prospective control study. A total of 725 patients with hypertensive basal ganglia cerebral haemorrhage admitted to department of neurosurgery from January 2001 to May 2007 were enrolled. They were diagnosed according to the diagnostic criteria set by the fourth national cerebrovascular disease conference in 1995. Haematoma volume was ＞ 50 mL. All patients were treated with craniotomy. And those with respiration and circulation failure, neurologic function deficit before the onset of the disease,major organ dysfunction, haemorrhagic disease and bleeding tendency or applied medicines affecting coagulation function excepted aspirin were excluded. The patients without use of aspirin before the onset of the disease were operated as the control group(group A), and there were 389 patients in group A.The patients with use of aspirin before the onset of the disease were randomly assigned to group B and C group,and there were 168 patients in group B or group C.The patients in group C received the frozen apheresis platelets. We counted different haematoma volume of relapsing haemorrhage after operation,death rate,ADL scores grades by 6 months follow-up survey in three groups. Quantitative data were expressed as mean ± standard deviation (-x ± s). The data were analyzed by using Chi-square test and Student's t test and rank sum test with SPSS 13.0 statistical package. A P value less than 0.05 indicated statisticals significance. Results Haematoma volume of relapsing haemorrhage was (40.59 + 20. 061 )mL, (53.21 ± 21.260) mL, (40.68 ± 19.517) mL in groups A, B, C,respectively. There was significant difference between group A and group B ( P ＜ 0.01 ), between group B and group C ( P ＜ 0.05), but there was no significant difference between group A and group C(P ＞ 0.05). ADL scores grades at 6-month follow-up was (67.04 ± 26. 176), (54.47 ± 29.403 ), (68.21 ± 25.254) in groups A, B, C, respectively. There was more significant difference between group A and group B, in ADL scores grades and the death rate between group B and group C (P ＜ 0.01), but there was no significant difference between group A and group C (P ＞ 0.05). Conclusions Aspirin can increase the occurrence rate of haemorrhage after operation, disablement and death in cerebral haemorrhage patients, but frozen apheresis platelets can reduce the occurrence rate.

Flow cytometry analysis of immunophenotype features and folate receptor expression of blasts in myelodysplastic syndromes / 中国基层医药

Objective To explore the immunophenotype features and folate receptor expression of blasts in patients with myelodysplastic syndromes(MDS).Methods Four-color flow cytometry using conventional and secondary gating strategies was used into analysis the immunophenotype features and folate receptor(FR) expression of blasts and CD+34 cells in bone marrow nucleated cells with MDS.The patients with acute myeloid leukemia-M2(AML-M2) were as positive control.Results with progression of MDS from RA/RAS,RAEB to RAEB-T,using conventional gating strategy,the proportion of CD+34 cells were gradually increased(P＜0.05).Moreover,the expression of HLA-DR,CDll7,CD13,CD33 were also gradually increased and the expression of CDl5 was gradually decreased(P＜0.05).Using secondary gating strategy,the expression of HLA-DR,CD117,CD13,CD33 on blasts were higher than those by conventional gating(P＜0.05).However,there was no significant difference(P＞0.05) in the expression of above mentioned antigens on CD+34 cells among different MDS subtypes.On the other hand,there were no expression of FR on blasts and cD+34 cells with different MDS subtypes.Conclusion With progression of MDS,the antigens of blasts surface change into more immature immunophenotype of medullary system.But these antigens abnormal expression only illustrates the increase of ascendant malignant clone quantity,it can not reflect the nature of the disease.Using flow cytometry technique can not detect whether or not FR expression on the blasts with MDS.