OBJECTIVE To investigate the effects of Setaria italica extract on improving insomnia model mice and to explore its potential mechanisms. METHODS The mice were randomly assigned into blank group， model group， positive control group （diazepam， 2.6 mg/kg）， and S. italica extract low-dose， medium-dose and high-dose groups （1.2， 2.4， 4.8 g/kg）， with 10 mice in each group. Except for the blank group， all other groups received intraperitoneal injection of para-chlorophenylalanine （PCPA） to establish the insomnia model. After modeling， the blank group and model group were given a constant volume of normal saline intragastrically， and administration groups were given relevant medicine intragastrically， with a volume of 0.01 mL/g， once a day， for 7 consecutive days. After the administration， the open-field test was conducted to observe the praxiological changes of mice， and to determine the levels of 5-hydroxytryptamine （5-HT） and 5-hydroxyindoleacetic acid （5-HTAA） in the hippocampal tissue， as well as the contents of 5-HT， brain-derived neurotrophic factor （BDNF）， interleukin-2 （IL-2）， IL-6， B-cell lymphoma-2 （Bcl- 2）， and Bcl-2-associated X protein （Bax） in the serum. The expression of phosphoinositide 3-kinase/protein kinase B/nuclear factor- κB （PI3K/Akt/NF-κB） signaling pathway related protein was determined in the hippocampus of mice. RESULTS Compared with the model group， the total exercise time of mice in S. italica extract high-dose group was significantly prolonged， but the total rest time was significantly shortened （P＜0.01）； the number of standing times and modification times were significantly reduced （P＜ 0.01）. The contents of 5-HT， BDNF， and Bcl-2 in serum， and Bcl-2/Bax were significantly increased， while the contents of IL-2， IL-6， and Bax were significantly reduced （P＜0.05 or P＜ 0.01）. The content of 5-HTAA in the hippocampal tissue and 202104010910029）；the phosphorylation levels of PI3K and Akt proteins were increased significantly， while the phosphorylation level of NF-κB p65 protein was decreased significantly （P＜0.05）.CONCLUSIONS High-dose of S. italica extract demonstrates significant therapeutic effects on insomnia in mice， and the mechanism of which may be associated with the regulation of PI3K/Akt/NF-κB signaling pathway.

Objective To investigate the role of RNA-binding motif protein X-linked(RBMX)in regulating the proliferation,migration,invasion and glycolysis in human bladder cancer cells.Methods A lentivirus vectors system and RNA interference technique were used to construct bladder cancer 1376 and UC-3 cell models with RBMX overexpression and knockdown,respectively,and successful cell modeling was verified using RT-qPCR and Western blotting.Proliferation and colony forming ability of the cells were evaluated using EdU assay and colony-forming assay,and cell migration and invasion abilities were determined using Transwell experiment.The expressions of glycolysis-related proteins M1 pyruvate kinase(PKM1)and M2 pyruvate kinase(PKM2)were detected using Western blotting.The effects of RBMX overexpression and knockdown on glycolysis in the bladder cancer cells were assessed using glucose and lactic acid detection kits.Results RT-qPCR and Western blotting confirmed successful construction of 1376 and UC-3 cell models with RBMX overexpression and knockdown.RBMX overexpression significantly inhibited the proliferation,clone formation,migration and invasion of bladder cancer cells,while RBMX knockdown produced the opposite effects.Western blotting results showed that RBMX overexpression increased the expression of PKM1 and decreased the expression of PKM2,while RBMX knockdown produced the opposite effects.Glucose consumption and lactate production levels were significantly lowered in the cells with RBMX overexpression(P<0.05)but increased significantly following RBMX knockdown(P<0.05).Conclusion RBMX overexpression inhibits bladder cancer progression and lowers glycolysis level in bladder cancer cells by downregulating PKM2 expression,suggesting the potential of RBMX as a molecular target for diagnosis and treatment of bladder cancer.

Objective To investigate the role of RNA-binding motif protein X-linked(RBMX)in regulating the proliferation,migration,invasion and glycolysis in human bladder cancer cells.Methods A lentivirus vectors system and RNA interference technique were used to construct bladder cancer 1376 and UC-3 cell models with RBMX overexpression and knockdown,respectively,and successful cell modeling was verified using RT-qPCR and Western blotting.Proliferation and colony forming ability of the cells were evaluated using EdU assay and colony-forming assay,and cell migration and invasion abilities were determined using Transwell experiment.The expressions of glycolysis-related proteins M1 pyruvate kinase(PKM1)and M2 pyruvate kinase(PKM2)were detected using Western blotting.The effects of RBMX overexpression and knockdown on glycolysis in the bladder cancer cells were assessed using glucose and lactic acid detection kits.Results RT-qPCR and Western blotting confirmed successful construction of 1376 and UC-3 cell models with RBMX overexpression and knockdown.RBMX overexpression significantly inhibited the proliferation,clone formation,migration and invasion of bladder cancer cells,while RBMX knockdown produced the opposite effects.Western blotting results showed that RBMX overexpression increased the expression of PKM1 and decreased the expression of PKM2,while RBMX knockdown produced the opposite effects.Glucose consumption and lactate production levels were significantly lowered in the cells with RBMX overexpression(P<0.05)but increased significantly following RBMX knockdown(P<0.05).Conclusion RBMX overexpression inhibits bladder cancer progression and lowers glycolysis level in bladder cancer cells by downregulating PKM2 expression,suggesting the potential of RBMX as a molecular target for diagnosis and treatment of bladder cancer.

Objective:To explore the effect of the developmental stages and quality on pregnancy outcome and birth outcome, and provide evidence for single blastocyst selection in frozen-thawed cycles.Methods:A retrospective cohort study analysis was performed on the data of patients with a total of 893 cycles who underwent single blastocyst transfer in frozen-thawed cycles in the Center for Reproductive Medicine, Qingyuan People's Hospital from January 2013 to June 2021. The cycles were divided into day 5 (D5) and day 6 (D6) groups according to the time of blastocyst formation. Then the two groups were divided into four subgroups according to the quality of blastocyst, namely, D5 good-quality embryo subgroup, D5 non-good-quality embryo subgroup, D6 good-quality embryo subgroup and D6 non-good-quality embryo subgroup. The general data, clinical outcomes and neonatal outcomes of each group were compared.Results:1) The clinical pregnancy rate [60.14% (332/552)], the implantation rate [60.14% (332/552)] and the live birth rate [47.64% (263/552)] in D5 group were significantly higher than those in D6 group [45.75% (156/341), 45.75% (156/341), 36.36% (124/341), all P<0.001], but there were no significant differences in body mass index, duration of infertility, intimal thickness of transplantation day and miscarriage rate between the two groups (all P>0.05). In addition, there were also no significant differences in birth weight, low birth weight rate, fetal macrosomia rate and male/female ratio (all P>0.05). 2) There were significant differences in clinical pregnancy rate [61.00% (294/482), 54.29% (38/70), 51.00% (127/249), 31.52% (29/92)] and live birth rate [48.96% (236/482), 38.57% (27/70), 41.37% (103/249), 22.83% (21/92)] among D5 good-quality embryo subgroup, D5 non-good-quality embryo subgroup, D6 good-quality embryo subgroup and D6 non-good-quality embryo subgroup (all P<0.001). D5 good-quality embryo subgroup had the highest clinical pregnancy rate and live birth rate, while D6 non-good-quality embryo subgroup had the lowest clinical pregnancy rate and live birth rate. There were also no significant differences in birth weight, fetal macrosomia rate and male/female ratio among the four subgroups (all P>0.05), while there was a significant difference in low birth weight rate [5.08% (12/236), 0 (0/27), 4.85% (5/103), 23.81% (5/21)] among the four subgroups ( P=0.014). 3) There were no significant differences in clinical pregnancy rate and live birth rate between D5 non-good-quality embryo subgroup and D6 good-quality embryo subgroup (all P>0.05). The clinical pregnancy rate and the live birth rate of 4BC in D5 were lower than those of 4AA, 4AB and 4BA in D6, while the miscarriage rate of 4BC in D5 was higher than that of 4AA, 4AB and 4BA in D6, but there were no significant differences (all P>0.05).The clinical pregnancy rate and the live birth rate of 4BC in D5 were higher than those of 4BB in D6, but there were no significant differences (all P>0.05). Conclusion:In the frozen-thawed cycle of single blastocyst transplantation, D5 good-quality blastocysts are preferred. When faced with D5 non-good-quality embryos and D6 good-quality embryos, the optimal choice was D6 4AA>D6 4BA>D6 4AB>D5 4BC>D6 4BB.

Objective:To explore the effect of the developmental stages and quality on pregnancy outcome and birth outcome, and provide evidence for single blastocyst selection in frozen-thawed cycles.Methods:A retrospective cohort study analysis was performed on the data of patients with a total of 893 cycles who underwent single blastocyst transfer in frozen-thawed cycles in the Center for Reproductive Medicine, Qingyuan People's Hospital from January 2013 to June 2021. The cycles were divided into day 5 (D5) and day 6 (D6) groups according to the time of blastocyst formation. Then the two groups were divided into four subgroups according to the quality of blastocyst, namely, D5 good-quality embryo subgroup, D5 non-good-quality embryo subgroup, D6 good-quality embryo subgroup and D6 non-good-quality embryo subgroup. The general data, clinical outcomes and neonatal outcomes of each group were compared.Results:1) The clinical pregnancy rate [60.14% (332/552)], the implantation rate [60.14% (332/552)] and the live birth rate [47.64% (263/552)] in D5 group were significantly higher than those in D6 group [45.75% (156/341), 45.75% (156/341), 36.36% (124/341), all P<0.001], but there were no significant differences in body mass index, duration of infertility, intimal thickness of transplantation day and miscarriage rate between the two groups (all P>0.05). In addition, there were also no significant differences in birth weight, low birth weight rate, fetal macrosomia rate and male/female ratio (all P>0.05). 2) There were significant differences in clinical pregnancy rate [61.00% (294/482), 54.29% (38/70), 51.00% (127/249), 31.52% (29/92)] and live birth rate [48.96% (236/482), 38.57% (27/70), 41.37% (103/249), 22.83% (21/92)] among D5 good-quality embryo subgroup, D5 non-good-quality embryo subgroup, D6 good-quality embryo subgroup and D6 non-good-quality embryo subgroup (all P<0.001). D5 good-quality embryo subgroup had the highest clinical pregnancy rate and live birth rate, while D6 non-good-quality embryo subgroup had the lowest clinical pregnancy rate and live birth rate. There were also no significant differences in birth weight, fetal macrosomia rate and male/female ratio among the four subgroups (all P>0.05), while there was a significant difference in low birth weight rate [5.08% (12/236), 0 (0/27), 4.85% (5/103), 23.81% (5/21)] among the four subgroups ( P=0.014). 3) There were no significant differences in clinical pregnancy rate and live birth rate between D5 non-good-quality embryo subgroup and D6 good-quality embryo subgroup (all P>0.05). The clinical pregnancy rate and the live birth rate of 4BC in D5 were lower than those of 4AA, 4AB and 4BA in D6, while the miscarriage rate of 4BC in D5 was higher than that of 4AA, 4AB and 4BA in D6, but there were no significant differences (all P>0.05).The clinical pregnancy rate and the live birth rate of 4BC in D5 were higher than those of 4BB in D6, but there were no significant differences (all P>0.05). Conclusion:In the frozen-thawed cycle of single blastocyst transplantation, D5 good-quality blastocysts are preferred. When faced with D5 non-good-quality embryos and D6 good-quality embryos, the optimal choice was D6 4AA>D6 4BA>D6 4AB>D5 4BC>D6 4BB.

OBJECTIVE To study the impr ovement effects of Dianxianqing granule on blood-brain barrier （BBB）injury in Alzheimer’s disease （AD）model mice by regulating NLR family pyrin domain containing 3（NLRP3）inflammasome signaling pathway. METHODS Totally 125 mice were randomly divided into sham operation group （n＝25）and modeling group （n＝100） by body weight. AD model was induced by intracerebroventricular injection of β-amyloid 25-35 in model group. Sham operation group was given normal saline with same method. The 100 model mice were randomly divided into model group ，Donepezil hydrochloride tablets group （positive control 1，1.3 mg/kg，i.g.），MCC950 group [positive control 2（selective NLRP 3 inhibitor），10 mg/kg，i.p.] and Dianxianqing granule group （12.48 g/kg by crude drug ，i.g.）by body weight ，with 25 mice in each group. Second day after modeling ，administration groups were given relevant medicine ，once a day ，for consecutive 21 d. Sham operation group and model group were given intragastric administration of water and intraperitoneal injection of normal saline. At last administration，the learning and memory ability was determined by Y maze test ，and blood-brain barrier permeability was measured by Evans blue leakage assay. The expressions of NLRP 3，anti-ionized calcium-binding adapter molecule 1（IBA-1），nuclear factor kappa B （NF-κB）p65，p53 upregulated modulator of apoptosis （PUMA），occludin（ocln），zonula occluden- 1（ZO-1）and claudin-5 （cldn5） in cerebral tissue were determined. RESULTS Compared with model group ， spontaneous alternate response rate ，protein expressions of ocln ，cldn5 lnzyxyqy2003@163.com and ZO- 1 in cerebral tissue were increased significantly in administration groups （P＜0.05 or P＜0.01）；Evans blue E-mail：jiadg2003@126.com content and protein expressions of NLRP 3，IBA-1，PUMA and NF-κB p65 in cerebral tissue were decreased significantly （P＜0.05 or P＜0.01）. CONCLUSIONS Dianxianqing granule can improve BBB injury of AD model m ice by inhibiting NLRP 3 inflammasome signaling pathway.

AIM: To study the effect of ligustrazine on the cardiacmyocyte lesion in rats with type 2 diabetes mellitus.METHODS: Male Wistar rats were injected with STZ via tail vein under high-glucose and high-fat feeding for 4 weeks to establish the animal model of type 2 diabetes mellitus.Ligustrazine at different doses was used to treat the diabetic rats.The body weight, blood glucose and the morphology of heart tissues were observed.The myocardial levels of IL-1β, IL-6 and TNF-α were detected by ELISA, and the protein expression of IKKβ and NF-κB in the myocardium was determined by Westeren blotting.RESULTS: Ligustrazine at high dose alleviated the body weight reduction and blood glucose elevation cause by diabetes, and reduced pro-inflammatory factors IL-1β, TNF-α and IL-6.Moreover, the protein expression of IKKβ and NF-κB was significant decreased by ligustrazine.CONCLUSION: Ligustrazine inhibits the myocardial inflammation caused by diabetes through anti-inflammatory pathway.

Objective To investigate the effects of ovarian induction with raloxifene(RAL)versus clomi-phene citrate(CC)on the endometrial receptivity in mouse endometrium during perimplantation period. Methods 48 female Kun-ming mouse were randomly divided into four groups in equal number:RAL 240 mg group,RAL 180 mg group,CC group,natural conception group(NC),all treated with ovulation induction after drug administration.Successfully mated female mouse were killed,and uterus samples were collected for HE stain-ing and immunohistochemistry. Results HE staining showed that the endometrial morphology in the RAL 180 mg group and RAL 240 mg group and NC group were better than that of CC group.The expressions of COX-2 and LP-AR3 in the RAL 180 mg group and RAL 240 mg group were similar to NC group,without significant difference among the three groups(P > 0.05). But in the CC group,it was statistically significantly lower than other three groups(P<0.05),indicating ovarian induction with RAL did not decrease the expressions of COX-2 and LPAR3 in endometrium. Conclusion The mechanism of ovulation induction with RAL is similar to CC,but RAL has fewer adverse effects on the endometrial receptivity compared with CC.

OBJECTIVETo investigate the value of serum beta-human chorionic gonadotropin (β-HCG) on days 9 (d9) and 11 (d11) after fresh embryo transfer (ET) and frozen-thawed embryo transfer (FET) in predicting the pregnancy outcomes.

METHODSA total of 227 fresh ET and FET cycles performed at the Center of Reproductive Medicine, Qingyuan People's Hospital between Dec, 2012 and Feb, 2014 were analyzed. The data of serum β-HCG levels on d9 and d11 after fresh ET and FET cycles and the pregnancy outcomes were reviewed, and the ROC curve was constructed to determine the optimal cut-off level of serum β-HCG level for predicting pregnancy outcomes.

RESULTSAccording to pregnancy outcomes, the cycles were divided into non-pregnancy group (group A), biochemical pregnancy group (group B), and clinical pregnancy group (group C). Significant differences were found between the mean serum β-HCG levels measured on d9 and d11 in all the 3 groups (P<0.01). The cycles in group C were further divided into 5 subgroups with single pregnancy (C1), twin pregnancy (C2), early abortion (C3), ectopic pregnancy (C4), or intrauterine pregnancy complicated with ectopic pregnancy group (C5), and all the 5 subgroups showed significant differences in β-HCG measurements between d9 and d11 (P<0.01). On d9 after ET and FET, the optimal cut-off level of serum β-HCG was 49.05 IU/L for predicting for clinical pregnancy, and was 105.15 IU/L for predicting twin pregnancy; the two corresponding cut-off levels of serum β-HCG on d11 was 51.2 IU/L and 241.75 IU/L, respectively.

CONCLUSIONThe absolute serum β-HCG level on d9 and d11 after fresh ET and FET allows an accurate diagnosis of pregnancy and helps in the prediction of the pregnancy outcomes.

Chorionic Gonadotropin, beta Subunit, Human ; blood ; Embryo Transfer ; Female ; Fertilization in Vitro ; Humans ; Pregnancy ; Pregnancy Outcome ; ROC Curve ; Retrospective Studies

Objective To investigate the value of serum beta-human chorionic gonadotropin (β-HCG) on days 9 (d9) and 11 (d11) after fresh embryo transfer (ET) and frozen-thawed embryo transfer (FET) in predicting the pregnancy outcomes. Methods A total of 227 fresh ET and FET cycles performed at the Center of Reproductive Medicine, Qingyuan People's Hospital between Dec, 2012 and Feb, 2014 were analyzed. The data of serumβ-HCG levels on d9 and d11 after fresh ET and FET cycles and the pregnancy outcomes were reviewed, and the ROC curve was constructed to determine the optimal cut-off level of serum β-HCG level for predicting pregnancy outcomes. Results According to pregnancy outcomes, the cycles were divided into non-pregnancy group (group A), biochemical pregnancy group (group B), and clinical pregnancy group (group C). Significant differences were found between the mean serumβ-HCG levels measured on d9 and d11 in all the 3 groups (P<0.01). The cycles in group C were further divided into 5 subgroups with single pregnancy (C1), twin pregnancy (C2), early abortion (C3), ectopic pregnancy (C4), or intrauterine pregnancy complicated with ectopic pregnancy group (C5), and all the 5 subgroups showed significant differences inβ-HCG measurements between d9 and d11 (P<0.01). On d9 after ET and FET, the optimal cut-off level of serumβ-HCG was 49.05 IU/L for predicting for clinical pregnancy, and was 105.15 IU/L for predicting twin pregnancy; the two corresponding cut-off levels of serum β-HCG on d11 was 51.2 IU/L and 241.75 IU/L, respectively. Conclusion The absolute serumβ-HCG level on d9 and d11 after fresh ET and FET allows an accurate diagnosis of pregnancy and helps in the prediction of the pregnancy outcomes.