Nrf2 expression, mitochondrial fission, and neuronal apoptosis in the prefrontal cortex of methamphetamine abusers and rats.

Methamphetamine (MA), a representative amphetamine-type stimulant, is one of the most abused drugs worldwide. Studies have shown that MA-induced neurotoxicity is strongly associated with oxidative stress and apoptosis. While nuclear factor E2-related factor 2 (Nrf2), an antioxidant transcription factor, is known to exert neuroprotective effects, its role in MA-induced dopaminergic neuronal apoptosis remains incompletely understood. In the present study, we explored the effects of MA on the expression levels of Nrf2, dynamin-related protein 1 (Drp1), mitofusin 1 (Mfn1), cytochrome c oxidase (Cyt-c), and cysteine aspartate-specific protease 3 (Caspase 3), as well as the correlations between Nrf2 and mitochondrial dynamics and apoptosis. Brain tissue from MA abusers was collected during autopsy procedures. An MA-dependent rat model was also established by intraperitoneal administration of MA (10 mg/kg daily) for 28 consecutive days, followed by conditioned place preference (CPP) testing. Based on immunohistochemical staining and western blot analysis, the protein expression levels of Nrf2 and Mfn1 showed a decreasing trend, while levels of Drp1, Cyt-c, and Caspase 3 showed an increasing trend in the cerebral prefrontal cortex of both MA abusers and MA-dependent rats. Notably, the expression of Nrf2 was positively associated with the expression of Mfn1, but negatively associated with the expression levels of Drp1, Cyt-c, and Caspase 3. These findings suggest that oxidative stress and mitochondrial fission contribute to neuronal apoptosis, with Nrf2 potentially playing a critical role in MA-induced neurotoxicity.

Triggering Receptor Expressed on Myeloid Cells 2 Deficiency Exacerbates Methamphetamine-Induced Activation of Microglia and Neuroinflammation.

Methamphetamine (METH) is a highly addictive psychostimulant and one of the most widely abused drugs worldwide. The continuous use of METH eventually leads to neurotoxicity and drug addiction. Studies have shown that neurotoxicity is strongly associated with METH-induced neuroinflammation, and microglia are the key drivers of neuroinflammation. Triggering receptor expressed on myeloid cells 2 (TREM2) is reported to play a key role in activation of microglia and neuroinflammation. Yet, the molecular mechanisms by which METH causes neuroinflammation and neurotoxicity remain elusive. In the current study, we investigated the role of TREM2 in neuroinflammation induced by METH in BV2 cells and the wild-type (WT) C57BL/6J mice, CX3CR1GFP/+ transgenic mice, and TREM2 knockout (KO) mice. Postmortem samples from the frontal cortex of humans with a history of METH use were also analyzed to determine the levels of TREM2, TLR4, IBA1, and IL-1ß. The expression levels of TREM2, TLR4, IBA1, IL-1ß, iNOS, and Arg-1 were then assessed in the BV2 cells and frontal cortex of mice and human METH users. Results revealed that the expression levels of TREM2, TLR4, IBA1, and IL-1ß were significantly elevated in METH-using individuals and BV2 cells. Microglia were clearly activated in the frontal cortex of WT C57BL/6 mice and CX3CR1GFP/+ transgenic mice, and the protein levels of IBA1, TREM2, TLR4, and IL-1ß were elevated in the METH-induced mouse models. Moreover, TREM2-KO mice showed further increased microglial activation, neuroinflammation, and excitotoxicity induced by METH. Thus, these findings suggest that TREM2 may be a target for regulating METH-induced neuroinflammation.

Nrf2 attenuates methamphetamine-induced myocardial injury by regulating oxidative stress and apoptosis in mice.

OBJECTIVES: Methamphetamine (MA) abuse is a serious social problem worldwide. Cardiovascular complications were the second leading cause of death among MA abusers. We aimed to clarify the effects of MA on myocardial injury, oxidative stress, and apoptosis in myocardial cells and to explore the potential mechanism of nuclear factor-erythroid factor 2-related factor 2 (Nrf2) in MA-induced oxidative stress and apoptosis. METHODS: An acute cardiac toxicity model of MA was established by intraperitoneal injection of MA (2 mg/kg) for 5 days. Nrf2 activation (by sulforaphane (SFN) 1 h before MA injection) and Nrf2 gene knockout were performed to explore the regulatory effects of Nrf2 on cardiac toxicity. RESULTS: The protein expressions of Nrf2 (p < .001) and heme oxygenase-1 (HO-1) were increased (p < .01), suggesting that MA activated the Nrf2/HO-1 pathway. In the MA group, cardiac injury score (p < .001) and cardiac troponin I (cTnI) protein expression increased (p < .01). Malondialdehyde (MDA) content increased (p < .001), superoxide dismutase (SOD) activity decreased (p < .05). Protein expressions of Caspase-3 (p < .001) and Bax (p < .001) increased, and Bcl-2 decreased (p < .001) as well. These changes were reversed by activation of Nrf2 but became more pronounced after Nrf2 knockout, suggested that the activation and knockout of Nrf2 attenuated and aggravated MA-induced myocardial injury, oxidative stress and apoptosis in myocardial cells, respectively. CONCLUSIONS: MA administration induced myocardial injury, oxidative stress, and apoptosis in mice. Nrf2 attenuated MA-induced myocardial injury by regulating oxidative stress and apoptosis, thus playing a protective role.

The effects of ketamine and methamphetamine on neurotransmitters, glutamate receptors, and conditioned place preference in rat.

Combined methamphetamine (MA) and ketamine (KET) abuse is a serious issue. At present, however, few studies have explored the mechanism underlying their combined addiction. We established a rat conditioned place preference (CPP) model. We investigated the role of dopamine (DA), 5-hydroxytryptamine (5-HT), monoamine oxidase (MAO), glutamate receptor 1 (GluR1), and glutamate receptor 2 (GluR2) in combined MA and KET addiction. The expression levels of DA, 5-HT, and MAO were detected by enzyme-linked immunosorbent assay (ELISA), and the expressions levels of GluR1 and GluR2 were detected by western blotting. Our results showed that MA and KET successfully induced CPP in rats respectively, and KET enhanced MA-induced CPP effects, although not significantly, and KET can reduce the MA-induced increase in DA, 5-HT, MAO and promoted the MA-induced increase in GluR1 and GluR2. Therefore, it suggested that DA, 5-HT, MAO, GluR1, and GluR2 expression may be involved in the mechanism underlying MA and KET-induced drug addiction in rats. Moreover, When MA and KET are used in combination, KET appears to play a dual addictive and anti-addictive role in the regulation of MA addiction.

Exercise attenuates neuronal degeneration in Parkinson's disease rat model by regulating the level of adenosine 2A receptor.

Parkinson's disease occurs due to loss of dopaminergic neurons, which alters the behavioural changes. The present study evaluates the effect of exercise on neurodegeneration against Parkinson's disease (PD) rat model and postulates its effect on novel molecular pathway. Rotenone was administered at 1 mg/kg s.c. every 48 h for 18 days for the in-duction of PD and exercise was given to rats for a period of 2 weeks after the confirmation of PD. Moreover, PD rats also received CGS 21680 (adenosine A2A receptor agonist, 0.5 mg/kg, i.p.) with exercise for a period of 2 weeks after confirmation of PD. The effect of exercise was assessed for motor and cognitive function in PD rats. The level of inflammatory cytokines and neurotransmitters was estimated in brain tissue of PD rats. Data of investigation reveal that exercise attenuates cognitive and motor function in PD rats, the exercise + CGS 21680 group shows reverse in the behavioural changes compared to exercise-treated PD rats. The level of inflammatory cytokines and neurochemical level ameliorated in the exercise-treated group compared to the PD group of rats, which is reversed in the exercise + CGS 21680 group. In conclusion, exercise protects neurodegeneration in PD rats by reducing aggregation of a-synuclein and activity of adenosine 2A receptor.

Effects of dynamic zero COVID-19 policy on anxiety status and lifestyle changes of pregnant women in rural South China: a survey-based analysis by propensity score matching method.

Introduction: The coronavirus disease 2019 (COVID-19) pandemic triggered a global public health crisis and has brought an unprecedented impact on pregnant women. The problems faced by pregnant women in the rural areas of China during the epidemic are different from those in urban areas. Although the epidemic situation in China has gradually improved, studying the impact of the previous dynamic zero COVID-19 policy on the anxiety status and lifestyle of pregnant women in rural areas of China, is still necessary. Methods: A cross-sectional survey of pregnant women in rural South China was conducted from September 2021 to June 2022.Using questionnaires, sociodemographic characteristics, anxiety status, physical activity, sleep quality, and dietary status of the population were collected. Using the propensity score matching method, the effect of the dynamic zero COVID-19 strategy on the anxiety status and lifestyle of pregnant women was analyzed. Results: Among the pregnant women in the policy group (n = 136) and the control group (n = 680), 25.7 and 22.4% had anxiety disorders, 83.1 and 84.7% had low or medium levels of physical activity, and 28.7 and 29.1% had sleep disorders, respectively. However, no significant difference (p > 0.05) was observed between the two groups. Compared with control group, the intake of fruit in the policy group increased significantly (p = 0.019), whereas that of aquatic products and eggs decreased significantly (p = 0.027). Both groups exhibited an unreasonable dietary structure and poor compliance with the Chinese dietary guidelines for pregnant women (p > 0.05). The proportion of pregnant women in the policy group, whose intake of stable food (p = 0.002), soybean, and nuts (p = 0.004) was less than the recommended amount, was significantly higher than that in the control group. Discussion: The dynamic zero COVID-19 strategy had little impact on the anxiety status, physical activity, and sleep disorders of pregnant women in the rural areas of South China. However, it affected their intake of certain food groups. Improving corresponding food supply and organized nutritional support should be addressed as a strategic approach to improve the health of pregnant women in rural South China during the pandemic.

6-Gingerol Improves In Vitro Porcine Embryo Development by Reducing Oxidative Stress.

6-Gingerol, the main active ingredient in ginger, exhibits a variety of biological activities, such as antioxidant, anti-inflammatory, and anticancer activities, and can affect cell development. However, the effects of 6-gingerol on mammalian reproductive processes, especially early embryonic development, are unclear. This study explored whether 6-gingerol can be used to improve the quality of in vitro-cultured porcine embryos. The results showed that 5 µM 6-gingerol significantly increased the blastocyst formation rates of porcine early embryos. 6-Gingerol attenuated intracellular reactive oxygen species accumulation and autophagy, increased intracellular glutathione levels, and increased mitochondrial activity. In addition, 6-gingerol upregulated NANOG, SRY-box transcription factor 2, cytochrome c oxidase subunit II, mechanistic target of rapamycin kinase, and RPTOR independent companion of MTOR complex 2 while downregulating Caspase 3, baculoviral IAP repeat containing 5, autophagy related 12, and Beclin 1. Most importantly, 6-gingerol significantly increased the levels of p-extracellular regulated protein kinase 1/2 while reducing the levels of p-c-Jun N-terminal kinase 1/2/3 and p-p38. These results indicate that 6-gingerol can promote the development of porcine early embryos in vitro.

Corrigendum to "Ginsenoside Rb1 attenuates methamphetamine (METH)-induced neurotoxicity through the NR2B/ERK/CREB/BDNF signalings in vitro and in vivo models" [J Ginseng Res 46 (2022) 426-434].

[This corrects the article DOI: 10.1016/j.jgr.2021.07.005.].

Correlation of lifestyle behaviors during pregnancy with postpartum depression status of puerpera in the rural areas of South China.

Background: Postpartum depression (PPD) is among the most common postpartum complications. Its prevalence is associated with strong regional variability. Women in rural areas of China have a high risk of PPD. The aim of this study was to investigate the PPD status of women in rural South China and explore the effects of modifiable lifestyle behaviors during pregnancy on their PPD status, thereby providing a scientific basis for the prevention and intervention of PPD in rural China. Methods: A cohort study was conducted on 261 women from four maternal health institutions situated in rural areas of Guangdong Province and the Guangxi Zhuang Autonomous Region from October 2021 to December 2022. The questionnaires were administered to these women to obtain data about sociodemographic characteristics, health literacy, physical activity during pregnancy, and sleep and dietary status during pregnancy, as well as depression status on the 42nd day after delivery. The lifestyle behaviors during pregnancy and the PPD status of the study population were analyzed. Multiple linear regression models were used to determine the correlation between lifestyle behaviors and PPD status. Path analysis was performed to explore the interaction between various lifestyle behaviors. Results: A total of 14.6% of women had a PPD status. Women who continued to work during pregnancy had an Edinburgh Postpartum Depression Scale (EPDS) score of 1.386 points higher than that of women who did not (В = 1.386, ß = 0.141, p = 0.029). For every 1-point increase in the infant feeding-related knowledge score and pregnancy diet diversity score, the EPDS score decreased by 0.188 and 0.484 points, respectively, and for every 1-point increase in the Pittsburgh sleep quality index score, the EPDS score increased by 0.288 points. Age was related to infant feeding-related knowledge (indirect path coefficient = 0.023). During pregnancy, sedentary time was correlated with sleep quality (indirect path coefficient = 0.031) and employment status (indirect path coefficient = 0.043). Conclusion: Employment status, infant feeding-related knowledge, sleep quality, and diet diversity during pregnancy directly influenced the PPD status, while age and sedentary time during pregnancy indirectly influenced the PPD status. Promoting healthy lifestyle behaviors, including reducing sedentary time, improving sleep quality, and increasing dietary diversity, may be effective in reducing PPD occurrence.

Cannabidiol attenuates methamphetamine-induced cardiac inflammatory response through the PKA/CREB pathway in rats.

Background: Methamphetamine (MA) abuse is a major global public health problem. However, it is not yet known whether cannabidiol (CBD) has protective effects on MA-induced cardiotoxicity. The present study investigated whether CBD has protective effects on MA-induced cardiac damage in rats via the protein kinase A/cyclic adenosine monophosphate (cAMP)-response element-binding protein (PKA/CREB) pathway. Methods: A total of 30 rats were randomly divided into 5 groups. The rats were administered MA (10 mg/kg) by intraperitoneal (IP) injection once a day for 4 weeks, and with CBD (40 or 80 mg/kg, IP) treatment 1 h before the MA injections. Morphological changes were determined using hematoxylin and eosin and Masson's trichrome staining. The serum levels of interleukin (IL)-6 and IL-10 were detected using enzyme-linked immunoassay kits. The protein expression levels of cardiac troponin I (cTnI), PKA, phospho-PKA (p-PKA), CREB, and phospho-CREB (p-CREB) in the myocardium were detected by Western blot analysis. Results: There was no significant difference in body weight among the groups. Heart weight and the heart-to-body weight ratio were higher in the MA group than the control group, while CBD (80 mg/kg) pretreatment (CBD80 + MA group) reduced the heart weight and the heart-to-body weight ratio compared to the MA group. The chronic administration of MA resulted in a cardiac inflammatory response, the progressive development of fibrosis, and necrosis, while CBD treatment attenuated these lesions. The protein expression levels of PKA, p-PKA, CREB, and p-CREB increased following MA administration, but significantly decreased with CBD treatment. These results indicate that chronic MA administration leads to cardiotoxicity, but these effects can be attenuated by CBD pretreatment. Conclusions: This study was the first to examine the protective effects of CBD on cardiotoxicity elicited by chronic MA exposure in rats. Our research suggests that CBD attenuates the cardiac inflammatory response induced by MA through the PKA/CREB pathway, and CBD may have potential clinical application in the treatment of MA-induced cardiotoxicity.

Cannabidiol prevents methamphetamine-induced neurotoxicity by modulating dopamine receptor D1-mediated calcium-dependent phosphorylation of methyl-CpG-binding protein 2.

In the past decade, methamphetamine (METH) abuse has sharply increased in the United States, East Asia, and Southeast Asia. METH abuse not only leads to serious drug dependence, but also produces irreversible neurotoxicity. Currently, there are no approved pharmacotherapies for the treatment of METH use disorders. Cannabidiol (CBD), a major non-psychoactive (and non-addictive) cannabinoid from the cannabis plant, shows neuroprotective, antioxidative, and anti-inflammatory properties under METH exposure. At present, however, the mechanisms underlying these properties remain unclear, which continues to hinder research on its therapeutic potential. In the current study, computational simulations showed that CBD and METH may directly bind to the dopamine receptor D1 (DRD1) via two overlapping binding sites. Moreover, CBD may compete with METH for the PHE-313 binding site. We also found that METH robustly induced apoptosis with activation of the caspase-8/caspase-3 cascade in-vitro and in-vivo, while CBD pretreatment prevented these changes. Furthermore, METH increased the expression of DRD1, phosphorylation of Methyl-CpG-binding protein 2 (MeCP2) at serine 421 (Ser421), and level of intracellular Ca2+ in-vitro and in-vivo, but these effects were blocked by CBD pretreatment. The DRD1 antagonist SCH23390 significantly prevented METH-induced apoptosis, MeCP2 phosphorylation, and Ca2+ overload in-vitro. In contrast, the DRD1 agonist SKF81297 markedly increased apoptosis, MeCP2 phosphorylation, and Ca2+ overload, which were blocked by CBD pretreatment in-vitro. These results indicate that CBD prevents METH-induced neurotoxicity by modulating DRD1-mediated phosphorylation of MeCP2 and Ca2+ signaling. This study suggests that CBD pretreatment may resist the effects of METH on DRD1 by competitive binding.

Ginsenoside Rb1 attenuates methamphetamine (METH)-induced neurotoxicity through the NR2B/ERK/CREB/BDNF signalings in vitro and in vivo models.

Aim: This study investigates the effects of ginsenoside Rb1 (GsRb1) on methamphetamine (METH)-induced toxicity in SH-SY5Y neuroblastoma cells and METH-induced conditioned place preference (CPP) in adult Sprague-Dawley rats. It also examines whether GsRb1 can regulate these effects through the NR2B/ERK/CREB/BDNF signaling pathways. Methods: SH-SY5Y cells were pretreated with GsRb1 (20 µM and 40 µM) for 1 h, followed by METH treatment (2 mM) for 24 h. Rats were treated with METH (2 mg/kg) or saline on alternating days for 10 days to allow CPP to be examined. GsRb1 (5, 10, and 20 mg/kg) was injected intraperitoneally 1 h before METH or saline. Western blot was used to examine the protein expression of NR2B, ERK, P-ERK, CREB, P-CREB, and BDNF in the SH-SY5Y cells and the rats' hippocampus, nucleus accumbens (NAc), and prefrontal cortex (PFC). Results: METH dose-dependently reduced the viability of SH-SY5Y cells. Pretreatment of cells with 40 µM of GsRb1 increased cell viability and reduced the expression of METH-induced NR2B, p-ERK, p-CREB and BDNF. GsRb1 also attenuated the expression of METH CPP in a dose-dependent manner in rats. Further, GsRb1 dose-dependently reduced the expression of METH-induced NR2B, p-ERK, p-CREB, and BDNF in the PFC, hippocampus, and NAc of rats. Conclusion: GsRb1 regulated METH-induced neurotoxicity in vitro and METH-induced CPP through the NR2B/ERK/CREB/BDNF regulatory pathway. GsRb1 could be a therapeutic target for treating METH-induced neurotoxicity or METH addiction.

KRAS Affects Adipogenic Differentiation by Regulating Autophagy and MAPK Activation in 3T3-L1 and C2C12 Cells.

Kirsten rat sarcoma 2 viral oncogene homolog (Kras) is a proto-oncogene that encodes the small GTPase transductor protein KRAS, which has previously been found to promote cytokine secretion, cell survival, and chemotaxis. However, its effects on preadipocyte differentiation and lipid accumulation are unclear. In this study, the effects of KRAS inhibition on proliferation, autophagy, and adipogenic differentiation as well as its potential mechanisms were analyzed in the 3T3-L1 and C2C12 cell lines. The results showed that KRAS was localized mainly in the nuclei of 3T3-L1 and C2C12 cells. Inhibition of KRAS altered mammalian target of rapamycin (Mtor), proliferating cell nuclear antigen (Pcna), Myc, peroxisome proliferator-activated receptor γ (PPARγ), CCAAT/enhancer binding protein beta (C/ebp-ß), diacylglycerol O-acyltransferase 1 (Dgat1), and stearoyl-coenzyme A desaturase 1 (Scd1) expression, thereby reducing cell proliferation capacity while inducing autophagy, enhancing differentiation of 3T3-L1 and C2C12 cells into mature adipocytes, and increasing adipogenesis and the capacity to store lipids. Moreover, during differentiation, KRAS inhibition reduced the levels of extracellular regulated protein kinases (ERK), c-Jun N-terminal kinase (JNK), p38, and phosphatidylinositol 3 kinase (PI3K) activation. These results show that KRAS has unique regulatory effects on cell proliferation, autophagy, adipogenic differentiation, and lipid accumulation.

Schisanhenol improves early porcine embryo development by regulating the phosphorylation level of MAPK.

Schisanhenol (SAL), a biphenyl cyclooctene-type lignin compound which can be extracted and isolated from many plants of the Schisandra family, exhibits a variety of biological activities including anti chronic cough, night sweating, thirst, diabetes, and obesity. However, its effects on the female reproductive system are unclear. Previous studies showed that SAL had potential antioxidant activity in heart, liver, and brain. Therefore, we hypothesized that SAL could improve porcine early development by reducing oxidative stress. The purpose of this study was to investigate the effects of SAL on preimplantation porcine embryos and the potential mechanisms. In this study, we analyzed the effects of SAL on embryo quality, reactive oxygen species (ROS) accumulation, mitochondrial function, cell proliferation and apoptosis, and the activation of MAPK pathway. The results showed that 10 µM SAL significantly increased the blastocyst formation rate, proliferation ability, and mitochondrial activity while reducing ROS accumulation and apoptosis level. During this process, the phosphorylation levels of ERK1/2, JNK1/2/3, and p38 were decreased. In summary, 10 µM SAL improves porcine preimplantation embryo development by reducing ROS accumulation.

Carnosic acid improves porcine early embryonic development by inhibiting the accumulation of reactive oxygen species.

Carnosic acid (CA), a natural catechol rosin diterpene, is used as an additive in animal feeds and human foods. However, the effects of CA on mammalian reproductive processes, especially early embryonic development, are unclear. In this study, we added CA to parthenogenetically activated porcine embryos in an in vitro culture medium to explore the influence of CA on apoptosis, proliferation, blastocyst formation, reactive oxygen species (ROS) levels, glutathione (GSH) levels, mitochondrial membrane potential, and embryonic development-related gene expression. The results showed that supplementation with 10 µM CA during in vitro culture significantly improved the cleavage rates, blastocyst formation rates, hatching rates, and total numbers of cells of parthenogenetically activated porcine embryos compared with no supplementation. More importantly, supplementation with CA also improved GSH levels and mitochondrial membrane potential, reduced natural ROS levels in blastomeres, upregulated Nanog, Sox2, Gata4, Cox2, Itga5, and Rictor expression, and downregulated Birc5 and Caspase3 expression. These results suggest that CA can improve early porcine embryonic development by regulating oxidative stress. This study elucidates the effects of CA on early embryonic development and their potential mechanisms, and provides new applications for improving the quality of in vitro-developed embryos.

Altered frequency of peripheral B-cell subsets and their correlation with disease activity in patients with systemic lupus erythematosus: A comprehensive analysis.

Alternations of peripheral B-cell subsets are closely related to disease activity in systemic lupus erythematosus (SLE) and may also predict the relapse of SLE. In this study, we aimed to comprehensively analyse the frequency of peripheral B-cell subsets, and their correlation with disease activity in patients with SLE. The results showed that for B-cell subsets in the antigen-independent differentiation stage, the frequency of the peripheral hematopoietic stem cell (HSC) subset in all patients with SLE was significantly higher than that of control patients. Surprisingly, several significant correlations were noted in newly diagnosed patients with SLE including a positive correlation in the frequency of the common lymphoid progenitor cell (CLP) with cholesterol serum levels. For B-cell subsets in the antigen-dependent differentiation stage, the frequency of naïve B-cell (N-B) subsets in all patients with SLE was significantly higher than that in the control patients. Moreover, the frequency of plasmablasts positively correlated with the SLEDAI score in the newly diagnosed patients. For memory B-cell (M-B) subtypes in the antigen-dependent differentiation stage, the frequency of the class-switched memory B-cell (CSM-B) subsets was positively correlated with the serum levels of complement C3. Notably, the frequency of the CSM-B subset also negatively correlated with the SLEDAI score, whereas the non-class-switched memory B-cell (NSM-B) subset was positively correlated with the serum levels of haemoglobin. Collectively, these findings may contribute to a better understanding of the role played by different B-cell subsets in the pathogenesis of SLE.

CPEB3 regulates the proliferation and apoptosis of bovine cumulus cells.

Cytoplasmic polyadenylation element-binding protein 3 (CPEB3) is a member of the Cytoplasmic polyadenylation element-binding family, which has been found to regulate the translation of dormant and masked mRNA in Xenopus oocytes and plays potential roles in regulating biological functions in cells and tissues. However, its role in cumulus cells is not clear. In this study, the mRNA expression of CPEB3 in bovine cumulus cells was inhibited with small interfering RNA. Cell cycle progression, proliferation, and apoptosis were measured after inhibition of CPEB3. Subsequently, changes in intracellular Reactive oxygen species content, mitochondrial membrane potential and expansion-related gene expression were examined. The results showed that after CPEB3 inhibition, cumulus cells had an abnormal cell cycle, the numbers of cells in the S and G2/M phases were significantly increased, cell proliferation was increased and apoptosis rates were decreased. These effects were likely due CPEB3 inhibition-induced decreases in intracellular Reactive oxygen species levels; increases in mitochondrial membrane potential; decreases in apoptosis; downregulation of CCNA, CCND, CCNE, CDK2, CDK4, CDK6, p21, and p27 mRNA expression; and upregulation of CCNB, CDK1, HAS2, PTGS2, PTX3, and CEBPB mRNA expression. Therefore, CPEB3 plays potential roles in regulating the biological and physiological functions of bovine cumulus cell.

[Effect of electroacupuncture on diaphragmatic function in patients with post-stroke tracheotomy].

OBJECTIVE: To observe the effect of electroacupuncture (EA) on diaphragmatic function based on conventional treatment in patients with post-stroke tracheotomy tube. METHODS: A total of 70 patients were randomized into an observation group (35 cases) and a control group (34 cases, 1 case dropped off). Internal medical basic treatment and breathing training were given in both groups. Besides, Xingnao Kaiqiao acupuncture was applied at Neiguan (PC 6), Shuigou (GV 26), Sanyinjiao (SP 6), Jiquan (HT 1), Chize (LU 5) and Weizhong (BL 40) in the control group, the needles were sustained for 30 min. On the basis of treatment in the control group, EA was applied at Tianding (LI 17), Fengchi (GB 20) and Jiaji (EX-B 2, C3-C5 ) in the observation group, with continuous wave, 10-20 Hz, 30 min each time. The treatment was given once a day, 6 times a week for 4 weeks in both groups. Before and after treatment, the diaphragmatic motility was measured by SonoSite ultrasound system (M-Turbo) in the two groups. RESULTS: Compared before treatment, the diaphragmatic motility after treatment was increased in both groups (P<0.01), and the improvement in the observation group was superior to the control group (P<0.01). CONCLUSION: Electroacupuncture can promote the recovery of diaphragmatic function in patients with post-stroke tracheotomy tube.

L-Arginine and N-carbamoylglutamic acid supplementation enhance young rabbit growth and immunity by regulating intestinal microbial community.

OBJECTIVE: An experiment was conducted to determine the effects of L-arginine (L-Arg) and N-carbamoylglutamic acid (NCG) on the growth, metabolism, immunity and community of cecal bacterial flora of weanling and young rabbits. METHODS: Eighteen normal-grade male weanling Japanese White Rabbits (JWR) were selected and randomly divided into 6 groups with or without L-Arg and NCG supplementation. The whole feeding process was divided into weanling stage (Day 37 to 65) and young stage (Day 66 to 85). The effects of L-Arg and NCG on the growth, metabolism, immunity and development of the ileum and jejunum were compared via nutrient metabolism experiments and histological assessment. The different communities of cecal bacterial flora affected by L-Arg and NCG were assessed using high-throughput sequencing technology and bioinformatics analysis. RESULTS: The addition of L-Arg and NCG were able to enhance the growth of weanling and young rabbit by increasing the nitrogen metabolism, protein efficiency ratio, and biological value, as well as feed intake, daily weight gain. Both L-Arg and NCG were able to increase the concentration of IgA, IgM, and IgG. NCG was superior to L-Arg in promoting intestinal villus development by increasing villus height and V/C index, reducing the crypt depth. The effects of L-Arg and NCG on the cecal bacterial flora were mainly concentrated in different genera, including Parabacteroides, Roseburia, dgA-11_gut_group, Alistipes, Bacteroides, and Ruminococcaceae_UCG-005. These bacteria function mainly in amino acid transport and metabolism, energy production and conversion, lipid transport and metabolism, recombination and repair, cell cycle control, cell division, and cell motility. CONCLUSION: L-Arg and NCG have promotional ability on the growth and immunity of weanling and young Japanese White Rabbits, as well as their effects on the jejunum and ileum villi. L-Arg and NCG have different effects in the promotion of nutrient utilization, relieving inflammation and enhancing adaptability through regulating microbial community.

Imperatorin improves in vitro porcine embryo development by reducing oxidative stress and autophagy.

Imperatorin (IMP), a furanocoumarin derivative with many biological properties and pharmacological activities, is widely used as an antibacterial, anti-inflammatory, antiviral, anticancer, cardiovascular and neuroprotective agent. The purpose of this study was to explore the effects of IMP on early embryo development in pigs as well as the potential mechanisms. Our results showed that IMP can enhance the developmental competence of porcine early embryos. Supplementation of in vitro culture medium with 40 µM IMP significantly increased the blastocyst rate and total cell number. At the same time, apoptosis of blastocysts was also significantly decreased in the supplemented group compared with the control group, in accordance with the subsequent results of FAS and CASP3 gene expression analysis. Furthermore, IMP attenuated intracellular reactive oxygen species (ROS) generation, increased fluorescein diacetate (FDA) and glutathione (GSH) levels. Importantly, IMP not only improved the activity of mitochondria but also inhibited the occurrence of autophagy. In addition, pluripotency-related genes (OCT4, NANOG, and SOX2) and a growth and metabolism regulatory gene (mTOR) were upregulated after IMP supplementation on Day 7. These results demonstrate that IMP exerts a beneficial effect on preimplantation embryo development by reducing oxidative stress and autophagy.