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.

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.

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.

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.

[Effect of "Tongyuan" acupuncture treatment on success rate of extubation in stroke patients undergoing tracheotomy].

OBJECTIVE: To observe the impact of "Tongyuan" (regulating the Governor Vessel to tranquilize mental activities, conducting qi back to its origin) acupuncture treatment on cough reflex, pulmonary infection and swallowing function in stroke patients undergoing tracheotomy. METHODS: Seventy-four cerebral stroke patients with tracheo-tomy were randomly allocated to scalp acupuncture and "Tongyuan" acupuncture groups (n=37 in each group). All the patients of the two groups received basic treatment, including treatment of primary diseases, routine nursing, respiratory muscle training, physical therapy, acupuncture of Chize (LU5), Quchi (LI11), Hegu (LI4), Weizhong (BL40), Zusanli (ST36). In addition, for patients of the scalp acupuncture group, the middle 2/5 of Dingnieqianxiexian (MS6), the middle 2/5 of Dingniehouxiexian (MS7) and Dingpangerxian (MS9) on the contralateral side of lesions were punctured with filiform needles. For patients of the "Tongyuan" acupuncture group, Baihui (GV20), Zhongwan (GV12), Guanyuan (CV4), Qihai (CV6), Tianshu (ST25), Feishu (BL13), Pishu (BL20) were needled. The treatment in both groups lasted 30 minutes each time, once daily, 6 times a week for 4 weeks. The cough reflex grading score (CRGS, assessed according to cough, independent expectoration, sputum suction), clinical pulmonary infection score (CPIS, assessed according to the body temperature, white blood cell count, airway secretion in 24 h, PaO2/FiO2, infiltration status, cultured pathogenic bacteria), and swallowing function were evaluated by a researcher who was blinded to the grouping and treatment procedures. The success rate of extubation was calculated in a week at the end of treatments. RESULTS: After the treatment, the CRGS and CPIS of both groups were apparently decreased in comparison with their own pre-treatment (P<0.05), suggesting an improvement of the cough reflex and pulmonary infection. Regarding the swallowing function, of the two 37 cases in the scalp acupuncture and "Tongyuan" acupuncture groups, 9(24.3%) and 19(51.4%) experienced marked improvement, 12(32.5%)and 14(37.8%) were effective, and 16(43.2%) and 4(10.8%)failed in the treatment, with the effective rate being 56.8% and 89.2%, respectively. In regard to the successful extubation, of the two 37 cases in the scalp acupuncture and "Tongyuan" acupuncture groups, 5(13.5%) and 11(29.8%)had a marked improvement, 10(27.0%)and 18(48.6%)were effective, and 22(59.5%)and 8(21.6%)failed, with the effective rate being 40.5% and 78.4%, respectively. The therapeutic effect of "Tongyuan" acupuncture was significantly superior to that of scalp acupuncture in decreasing CRGS and CPIS, and in improving the swallowing function and raising the rate of successful extubation (P<0.05). CONCLUSION: "Tongyuan" acupuncture treatment can improve cough reflex to remove sputum, reduce pulmonary infection, enhance swallowing function, and raise the success rate of extubation in stroke patients, which is better than scalp acupuncture.

COL1A1 affects apoptosis by regulating oxidative stress and autophagy in bovine cumulus cells.

The collagen type I alpha 1 chain (COL1A1), as a major component of extracellular matrix, plays a potential role in the growth and development of bovine follicles. However, its specific role in bovine cumulus cells remains unclear. In this study, we examined apoptosis, the cell cycle and reactive oxygen species after inhibition of COL1A1 expression by siRNA in bovine cumulus cells. Cell proliferation was measured by CCK-8, and mitochondrial membrane potential was detected by fluorescence intensities of JC-1 staining. Moreover, cell autophagy was detected by immunofluorescence, and cell migration was detected by a cell scratch assay. Lactic acid and cholesterol concentration were measured to evaluate the glucose utilization and cholesterol synthesis activity in cumulus cell by optical density detection method. RT-qPCR and Western blot analysis were used to measure changes in key gene expression. The results showed that cumulus cells were found to have an abnormal cell cycle, and the numbers of cells in S phase were significantly reduced, accompanied by decreases in cholesterol synthesis, and cell proliferation ability and an increase in apoptosis rate with siRNA-COL1A1 treatment. These findings were likely due to inhibition of COL1A1 resulting in high levels of ROS in the cells, a decrease in mitochondrial membrane potential, an increase in intracellular autophagy, activation of the apoptotic pathway, and a decrease in lactic acid conversion ability. COL1A1 plays an important role in regulating the physiological and biological functions of bovine cumulus cells.

Synonymous mutation adenomatous polyposis coliΔ486s affects exon splicing and may predispose patients to adenomatous polyposis coli/mutY DNA glycosylase mutation­negative familial adenomatous polyposis.

Familial adenomatous polyposis (FAP) is an autosomal dominant­inherited colorectal cancer. Recent advances in genetics have indicated that the majority of patients with FAP carry germline mutations of the adenomatous polyposis coli (APC) and mutY DNA glycosylase (MUTYH) genes. However, a large subset of families with a history of FAP have undetectable pathogenic alterations, termed APC/MUTYH mutation­negative FAP. To investigate the germline mutations in the APC and MUTYH genes in Chinese patients with FAP, 13 unrelated patients were enrolled. Through genetic sequencing, four known pathogenic alterations (Lys1061LysfsTer2, Glu1309AspfsTer4, Arg283Ter and Ser1196Ter) of APC and two novel disease­associated pathogenic mutations (Tyr152Ter and Ter522Gly) in MUTYH were identified in six individuals. For samples that did not present with pathogenic alterations, the functional effects of missense, synonymous and intronic mutations were analyzed using bioinformatics tools and databases. Bioinformatics prediction suggested that the synonymous mutation Tyr486Tyr in APC (APC∆486s) was likely a disease­causing polymorphism and may have induced the exon skipping of APC. A hybrid mini­gene assay was performed, which confirmed that the synonymous single nucleotide polymorphism APC∆486s induced major splicing defects with skipping of exon 12 in APC. The data of the present study suggested that the synonymous polymorphism APC∆486s was a potential pathogenic alteration that predisposed APC/MUTYH mutation­negative patients to FAP.