Objective To evaluate the application value of the blastocysts derived from non-pronucleus (0PN) zygotes by the good quality blastocyst formation rate and the clinical outcomes of frozen-thawed blastocyst transfers. Methods The good quality blastocyst formation rate derived from 0PN zygotes was compared with that derived from2 pronucleus(2PN)zygotes in in vitro fertilization(IVF)or intracytoplasmic sperm injection (ICSI) cycles from January 2015 to December 2016. In addition, the clinical pregnancy, embryo implantation and live birth rates of frozen-thawed blastocyst transfers with blastocysts derived from 0PN and 2PN zygotes were analyzed on corresponding dates. Results (1)In IVF cycles, the high quality blastocysts formation rate of 2PN embryos was significantly higher than that of 0PN (46.64% versus 42.42%, P<0.01). In ICSI cycles, the high quality blastocysts formation rate of 2PN embryos was markedly higher than that of 0PN(41.96% versus 21.73%, P<0.01).(2)In frozen-thawed embryo transfer cycles for IVF, the clinical pregnancy, implantation and live birth rates of D5 0PN blastocysts were significantly higher than those of D6 2PN(52.64% versus 46.78%, 49.91% versus 41.20%, 46.54% versus 39.56%, all P<0.05), however, the abortion and newborn abnormal rates of D5 0PN blastocysts were lower than those of D6 2PN blastocysts(17.37% versus 23.36%, 1.31% versus 4.21%, both P<0.05); the clinical pregnancy, implantation and livebirth rates of D5 2PN blastocysts were significantly higher than those of D5 0PN(59.73% versus 52.64%, 55.95% versus 49.91%, 53.03% versus 46.54%, all P<0.05), but newborn abnormal rate was a little higher than that of D5 0PN(3.90% versus 1.31%, P<0.05);the clinical pregnancy, implantation and live birth rates of D5 2PN blastocysts were significantly higher than those of D6 2PN(59.73% versus 46.78%, 55.95% versus 41.20%, 53.03% versus 39.56%, all P<0.05), and the abortion rate of D5 2PN blastocysts was lower than that of D6 2PN blastocysts(18.23% versus 23.36%, P<0.05). Conclusions Although the blastocysts derived from 0PN could be transffered, the blastocysts derived from 2PN zygotes are preferred in all cycles. In IVF cycles, the good quality blastocysts derived from 2PN or 0PN zygotes will be transferred.

Functional cognitive disorder (FCD) refers to complaints of persistent problematic cognitive decline, which is inconsistent between self-reported symptoms and daily function and/or neuropsychological test results, and the symptoms lasted for at least six months without obvious progress. Poor ability to reflect on and monitor cognitive processes has been suggested as a key mechanism underlying the disorder. In this review, the concept, research status, clinical manifestations and diagnosis of FCD were systematically examined, which is helpful to identify the subjective cognitive decline caused by non-degenerative diseases and conduct individualized intervention treatment.

Objective To evaluate the relationship between the hippocampal neuron-protective mechanism of hydrogen in a rat model of oxygen-glucose deprivation and restoration(OGD/R)and mito-chondrial autophagy.Methods Hippocampal neurons isolated from healthy Sprague-Dawley rats(24 h af-ter birth)were cultured in vitro,seeded in polylysine-coated 6-well plates at a density of 7×105 cells/well and then divided into 5 groups(n＝30 each)using a random number table method: control group(C group),OGD/R group,OGD/R+H2 group,OGD/R plus 3-methyladenine(3-MA)group(OGD/R+3-MA group),and OGD/R plus H2 plus 3-MA group(OGD/R+H2+3-MA group).The cells were cultured for 24 h in normal culture atmosphere(75％N2-20％O2-5％CO2)in group C,and cells were subjected to oxygen-glucose deprivation for 2 h followed by O2-glucose supply for 24 h to establish the model of OGD/R injury in OGD/R,OGD/R+H2,OGD/R+3-MA and OGD/R+H2+3-MA groups.The cells were cultured for 24 h in a hydrogen-rich incubator(60％H2-10％O2-5％CO2-25％N2)after establishing the model in group OGD/R+H2.Autophagy inhibitor 3-MA 10 mmol/L was added,and then cultured for 24 h in normal culture atmosphere after establishing the model in group OGD/R+3-MA.Autophagy inhibitor 3-MA 10 mmol/L was added,and then cultured for 24 h in hydrogen-rich incubator after establishing the model in group OGD/R+H2+3-MA.The cell survival rate was measured using MTT assay.DCFH-DA fluorescent probe was applied for determination of reactive oxygen species(ROS)activity.The mitochondrial membrane potential was measured using a JC-10 assay kit.The neuronal apoptosis was detected by flow cytometry,and apoptosis rate was calculated.The expression of mitophagy-related protein microtubule-associated protein 1 light chain 3(LC3),PINK1 and Parkin was determined by Western blot,and LC3Ⅱ/LC3Ⅰ ratio was calculated.Results Compared with group C,the cell survival rate and MMP were significantly decreased,the apop-tosis rate and ROS activity were increased,and the expression of PINK1 and Parkin and LC3Ⅱ/LC3Ⅰrati-o were increased in OGD/R and OGD/R+H2 groups(P＜0.05).Compared with group OGD/R,the cell survival rate and MMP were significantly increased,the apoptosis rate and ROS activity were decreased,and the expression of PINK1 and Parkin and LC3Ⅱ/LC3Ⅰ ratio were increased in group OGD/R+H2(P＜0.05),and the cell survival rate and MMP were significantly decreased,the apoptosis rate and ROS activ-ity were increased,and the expression of PINK1 and Parkin and LC3Ⅱ/LC3Ⅰ ratio were decreased in group OGD/R+3-MA(P＜0.05).Compared with group OGD/R+H2,the cell survival rate and MMP were significantly decreased,the apoptosis rate and ROS activity were increased,and the expression of PINK1 and Parkin and LC3Ⅱ/LC3Ⅰ ratio were decreased in OGD/R+3-MA and OGD/R+H2+3-MA groups(P＜0.05).Conclusion Hippocampal neuron-protective mechanism of hydrogen against OGDR injury is relat-ed to promoting mitochondrial autophagy in rats.

Objective: To evaluate the relationship between the hippocampal neuron-protective mechanism of hydrogen in a rat model of oxygen-glucose deprivation and restoration (OGD/R) and mitochondrial autophagy. Methods: Hippocampal neurons isolated from healthy Sprague-Dawley rats (24 h after birth) were cultured in vitro, seeded in polylysine-coated 6-well plates at a density of 7×10⁵ cells/well and then divided into 5 groups (n=30 each) using a random number table method: control group (C group), OGD/R group, OGD/R+ H₂ group, OGD/R plus 3-methyladenine (3-MA) group (OGD/R+ 3-MA group), and OGD/R plus H₂ plus 3-MA group (OGD/R+ H₂+ 3-MA group). The cells were cultured for 24 h in normal culture atmosphere (75%N₂-20%O₂-5%CO₂) in group C, and cells were subjected to oxygen-glucose deprivation for 2 h followed by O₂-glucose supply for 24 h to establish the model of OGD/R injury in OGD/R, OGD/R+ H₂, OGD/R+ 3-MA and OGD/R+ H₂+ 3-MA groups.The cells were cultured for 24 h in a hydrogen-rich incubator (60% H₂-10% O₂-5% CO₂-25% N₂) after establishing the model in group OGD/R+ H₂.Autophagy inhibitor 3-MA 10 mmol/L was added, and then cultured for 24 h in normal culture atmosphere after establishing the model in group OGD/R+ 3-MA.Autophagy inhibitor 3-MA 10 mmol/L was added, and then cultured for 24 h in hydrogen-rich incubator after establishing the model in group OGD/R+ H₂+ 3-MA.The cell survival rate was measured using MTT assay.DCFH-DA fluorescent probe was applied for determination of reactive oxygen species (ROS) activity.The mitochondrial membrane potential was measured using a JC-10 assay kit.The neuronal apoptosis was detected by flow cytometry, and apoptosis rate was calculated.The expression of mitophagy-related protein microtubule-associated protein 1 light chain 3 (LC3), PINK1 and Parkin was determined by Western blot, and LC3Ⅱ/LC3Ⅰ ratio was calculated. Results: Compared with group C, the cell survival rate and MMP were significantly decreased, the apoptosis rate and ROS activity were increased, and the expression of PINK1 and Parkin and LC3Ⅱ/LC3Ⅰ ratio were increased in OGD/R and OGD/R+ H₂ groups (P<0.05). Compared with group OGD/R, the cell survival rate and MMP were significantly increased, the apoptosis rate and ROS activity were decreased, and the expression of PINK1 and Parkin and LC3Ⅱ/LC3Ⅰ ratio were increased in group OGD/R+ H₂(P<0.05), and the cell survival rate and MMP were significantly decreased, the apoptosis rate and ROS activity were increased, and the expression of PINK1 and Parkin and LC3Ⅱ/LC3Ⅰ ratio were decreased in group OGD/R+ 3-MA (P<0.05). Compared with group OGD/R+ H₂, the cell survival rate and MMP were significantly decreased, the apoptosis rate and ROS activity were increased, and the expression of PINK1 and Parkin and LC3Ⅱ/LC3Ⅰ ratio were decreased in OGD/R+ 3-MA and OGD/R+ H₂+ 3-MA groups (P<0.05). Conclusion Hippocampal neuron-protective mechanism of hydrogen against OGDR injury is related to promoting mitochondrial autophagy in rats.

Neuropathic pain is a chronic disease that severely afflicts the life and emotional status of patients, but currently available treatments are often ineffective. Novel therapeutic targets for the alleviation of neuropathic pain are urgently needed. Rhodojaponin VI, a grayanotoxin from Rhododendron molle, showed remarkable antinociceptive efficacy in models of neuropathic pain, but its biotargets and mechanisms are unknown. Given the reversible action of rhodojaponin VI and the narrow range over which its structure can be modified, we perforwmed thermal proteome profiling of the rat dorsal root ganglion to determine the protein target of rhodojaponin VI. N-Ethylmaleimide-sensitive fusion (NSF) was confirmed as the key target of rhodojaponin VI through biological and biophysical experiments. Functional validation showed for the first time that NSF facilitated trafficking of the Cav2.2 channel to induce an increase in Ca²⁺ current intensity, whereas rhodojaponin VI reversed the effects of NSF. In conclusion, rhodojaponin VI represents a unique class of analgesic natural products targeting Cav2.2 channels via NSF.