Electroacupuncture alleviates neurological function via activating the Yap-OPA1 axis and mitochondrial fusion in rats with cerebral ischemia-reperfusion injury. / 电针通过Yesç›¸å ³è›‹ç™½-视神经萎缩蛋白1ä¿¡å·è½´æ¿€æ´»çº¿ç²’ä½“èžåˆåŠŸèƒ½å‡è½»å¤§é¼ è„‘ç¼ºè¡€å†çŒæ³¨æŸä¼¤çš„æœºåˆ¶ç ”ç©¶.

OBJECTIVES: To investigate the mechanism of electroacupuncture (EA) in alleviating cerebral ische-mia injury by activating the Yap-OPA1 signaling axis. METHODS: A total of 48 male SD rats were used in the present study. The focal CIRI model was established by occlusion of the middle cerebral artery and reperfusion (MCAO/R), followed by dividing the CIRI rats into model group, EA group and EA+Ver (Verteporfin, Yap antagonist) group (n=12 in each group). And another 12 normal rats were used as the sham operation group. For rats of the EA group, EA (4 Hz/20 Hz, 0.5 mA) was applied to "Baihui"(GV20) and "Shenting"(GV24) for 20 min, once daily for 7 days. The neurological deficit score (0 to 4 points) was given according to Longa's method. The infarct volume of rats in each group was assessed by TTC method, and the expression levels of Yes associated protein (Yap), Optic atrophy protein 1 (OPA1), mitofusin 1 (Mfn1), mitofusin 2 (Mfn2) proteins and mRNAs in cerebral cortex of infarcted side, as well as Bax (proapoptotic factor) and Bcl-1 (anti-apoptotic protein) proteins were detected by Westernblot, and real-time PCR, and the immunoactivity of Yap and OPA1 was detected by immunofluorescent staining. RESULTS: After modeling, the infarct volume, neurological deficit score and the expression of Bax were significantly increased (P<0.01), while the mRNA and protein expressions of Yap, OPA1, Mfn2, Mfn1, and Bcl-2 were significantly down-regulated in the model group relevant to the sham operation group (P<0.01, P<0.05). Compared with the model group, the neurological deficit score, infarct volume and the expression of Bax were significantly decreased (P<0.01), while the expression levels of Yap, OPA1, Mfn2, Mfn1 proteins and mRNAs and Bcl-2 protein, Yap and OPA1 immunofluorescence intensify were considerably up-regulated in the EA group (P<0.01, P<0.05). Following administration of Ver, the effects of EA in down-regulating the neurological score, infarct volume, and Bax expression and up-regulating the expressions of Yap, OPA1, Mfn1, Mfn2 proteins and mRNAs and Yap and OPA1 immunofluorescence intensify were eliminated. CONCLUSIONS: EA of GV20 and GV24 can improve the neurological function in rats with CIRI, which may be associated with its functions in activating mitochondrial fusion function and up-regulating Yap-OPA1 signaling axis.

[Electroacupuncture alleviates cerebral ischemia injury in rats by regulating melatonin-NLRP3 and inhibiting pyroptosis].

OBJECTIVE: To investigate the mechanism of electroacupuncture in alleviating cerebral ischemia injury in cerebral ischemia-reperfusion rats by regulating melatonin - NOD-like receptor protein 3 (NLRP3) mediated pyroptosis. METHODS: A total of 48 SD rats were randomly divided into sham operation group, model group, electroacupuncture (EA) group and EA +Luz group, with 12 rats in each group. The focal cerebral ischemia-reperfusion injury model was established by middle cerebral artery embolization. Rats of the EA group was treated with EA stimulation (4 Hz/20 Hz, 0.5 mA,20 min) at "Baihui" (GV20) and "Shenting" (GV24) once a day for 7 consecutive days; rats of EA+Luz group were given the same EA treatment and intraperitoneally administered melatonin receptor antagonist (luzindole, 30 mg/kg), once a day for 7 consecutive days. The neurological impairment was evaluated by Zea Longa score. The level of serum melatonin content at 12:00 and 24:00 was detected by ELISA. The percentage of cerebral infarction volume was evaluated by MRI of small animals. The apoptosis rate of nerve cells in cerebral cortex of infarct side was detected by TUNEL staining. The activation of microglia cells was detected by immunofluorescence staining. The expression levels of pyroptosis-related proteins NLRP3, Caspase-1 and interleukin (IL) -1ß were detected by Western blot. RESULTS: Compared with the sham operation group, the neural function score was significantly increased (P<0.01); the melatonin content was significantly decreased at 24:00 (P<0.01); the percentage of cerebral infarction volume, apoptosis rate of nerve cells in cerebral cortex area of infarction side, the expressions of NLRP3, Caspase-1 and IL-1ß proteins were significantly increased (P<0.01); and microglia cells were significantly activated in the model group.Compared with the model and EA +Luz groups, the nerve function score was significantly decreased (P<0.05); the percentage of cerebral infarction volume, the nerve cell apoptosis rate, the activation level of microglia cells, the expression levels of NLRP3, Caspase-1 and IL-1ß were significantly decreased (P<0.01, P<0.05) in the EA group. Compared with the model and EA+Luz groups, the melatonin content at 24:00 was significantly increased (P<0.01, P<0.05) in the EA group. CONCLUSION: EA at GV20 and GV24 can reduce the neurolo-gical injury in cerebral ischemia reperfusion model rats, which may be related to regulating the expression of endogenous melatonin, inhibiting cell scorchification and reducing cerebral ischemia injury.

[Sagittaria sagittifolia polysaccharides regulates Nrf2/HO-1 to relieve liver injury caused by multiple heavy metals in vivo and in vitro].

This study explored whether Sagittaria sagittifolia polysaccharides(SSP) activates the nuclear factor erythroid-2-related factor2(Nrf2)/heme oxygenase-1(HO-1) signaling pathway to protect against liver damage jointly induced by multiple heavy metals. First, based on the proportion of dietary intake of six heavy metals in rice available in Beijing market, a heavy metal mixture was prepared for inducing mouse liver injury and HepG2 cell injury. Forty male Kunming mice were divided into five groups: control group, model group, glutathione positive control group, and low-and high-dose SSP groups, with eight mice in each group. After 30 days of intragastric administration, the liver injury in mice was observed by HE staining. In the in vitro experiment, MTT assay was conducted to detect the effects of SSP at 0.25, 0.5, 1, and 2 mg·mL~(-1) on HepG2 cell survival at different time points. The content of alanine transaminase(ALT) and aspartate aminotransferase(AST) in the 48-h cell culture fluid was measured using micro-plate cultivation method, followed by the detection of the change in reactive oxygen species(ROS) content by flow cytometry. The mRNA expression levels of Nrf2 and HO-1 in cells were determined by RT-PCR, and their protein expression by Western blot. HE staining results showed that compared with the model group, the SSP administration groups exhibited significantly alleviated inflammatory cell infiltration and fatty infiltration in the liver, with better outcomes observed in the high-dose SSP group. In the in vitro MTT assay, compared with the model group, SSP at four concentrations all significantly increased the cell survival rate, decreased the ALT, AST, and ROS content(P<0.05), and down-regulated Nrf2 and HO-1 mRNA and protein expression(P<0.05). SSP significantly improves inflammatory infiltration in the liver tissue of mice exposed to a variety of heavy metals and corrects the liver fat degeneration, which may be related to its regulation of the Nrf2/HO-1 signaling pathway, reduction of ROS, and alleviation of oxidative damage.

Sagittaria sagittifolia polysaccharide protects against six-heavy-metal-induced hepatic injury associated with the activation of Nrf2 pathway to regulate oxidative stress and apoptosis.

The hepatic protective role of Sagittaria sagittifolia polysaccharide (SSP) and its possible mechanism were discussed in mice and L02 hepatocytes injured by heavy metals mixture of Cd + Cr (VI) + Pb + Mn + Zn + Cu. After 30-day intervention, blood and liver samples were collected for the relevant assessments. Methyl thiazolyl tetrazolium (MTT) assay showed 24 h was the best protecting point and the SSP protection at 1 mg/mL was strongest in L02 hepatocytes. SSP can alleviated hepatic injury, as evidenced by significantly decreased the activities of aspartate aminotransferase (AST), alanine aminotransferase (ALT), and the malondialdehyde (MDA) content, also increased the superoxide dismutase (SOD) activity and glutathione (GSH), total sulphydryl (T-SH) contents. SSP effectively reduced pathological damage of mice and accumulation of heavy metals in liver, as well as decreased the level of reactive oxygen species (ROS) in L02 hepatocytes. After SSP treatment, the protein expressions or gene transcription of nuclear factor erythroid 2-related factor 2 (Nrf2), NAD(P)H dehydrogenase, quinone 1 (NQO1) and heme oxygenase1 (HO-1) decreased in L02. The protein expression of Nrf2 and NQO1 were increased while HO-1 was decreased in liver. Besides, SSP can attenuates apoptosis through reducing the protein expression of Bcl-2-associated X protein (Bax) and caspase-3, and increasing B-cell lymphoma gene 2 (Bcl-2) and B-cell lymphoma-extra large (Bcl-xl). SSP protects against six-heavy-metal-induced hepatic injury in mice and L02 hepatocytes. Supported by Nrf2 gene silencing, the mechanisms may correlate with activating Nrf2 pathway to mitigate oxidative stress and apoptosis.

Analysis of the Correlation Between the Ratio of Monocytes to High-Density Lipoprotein Cholesterol and in-Stent Restenosis in Patients with Premature Coronary Heart Disease.

BACKGROUND: High-density lipoprotein cholesterol (HDL-C) and monocytes are associated with coronary artery disease, and the ratio of monocytes to high-density lipoprotein (MHR) is associated with long-term adverse outcomes and the recurrence of atrial fibrillation. Currently, the trend of coronary heart disease proned to young people is becoming prominent. However, the relationship between MHR and in-stent restenosis (ISR) in patients with premature coronary heart disease (PCHD) has not been investigated. Therefore, we aimed to assess the relationship between MHR and ISR in patients with PCHD. METHODS: We retrospectively included 257 patients (men ≤ 55 years old, women ≤ 65 years old) with PCHD who underwent drug-eluting stent implantation and follow-up coronary angiography at the First Affiliated Hospital of Zhengzhou University from September 2016 to September 2019. Patients were divided into ISR and non-ISR groups depending on their follow-up coronary angiography results. Relative clinical information was recorded and analyzed. A receiver operating characteristic curve analysis was used to determine the optimum pre-procedural MHR cutoff value to predict ISR. RESULTS: Logistic regression analysis showed that MHR, smoking history, and fibrinogen were independent risk factors for ISR in patients with PCHD. The area under the receiver operating characteristic curve (AUC) of MHR was 0.750 (95% confidence interval, 0.695-0.820; P < .001), the cutoff value was 546.88, and the specificity and sensitivity were 65.2% and 78%, while the AUC of monocytes was 0.631 (95% confidence interval, 0.638-0.794; P < .001), the cutoff value was 590, and the specificity and sensitivity were 77.1% and 60.0%. CONCLUSION: MHR is an independent risk factor for ISR in patients with PCHD and showed a certain predictive value.

Protoplast fusion enhances lignocellulolytic enzyme activities in Trichoderma reesei.

Protoplast fusion was used to obtain a higher production of lignocellulolytic enzymes with protoplast fusion in Trichoderma reesei. The fusant strain T. reesei JL6 was obtained from protoplast fusion from T. reesei strains QM9414, MCG77, and Rut C-30. Filter paper activity of T. reesei JL6 increased by 18% compared with that of Rut C-30. ß-Glucosidase, hemicellulase and pectinase activities of T. reesei JL6 were also higher. The former activity was 0.39 Uml(-1), while those of QM9414, MCG77, and Rut C-30 were 0.13, 0.11, and 0.16 Uml(-1), respectively. Pectinase and hemicellulase activities of JL6 were 5.4 and 15.6 Uml(-1), respectively, which were slightly higher than those of the parents. The effects of corn stover and wheat bran carbon sources on the cellulase production and growth curve of T. reesei JL6 were also investigated.

[Correlation between CTLA-4 gene polymorphism and systemic lupus erythematosus: a meta-analysis].

AIM: To determine the correlation between CTLA-4 gene polymorphism and systemic lupus erythematosus (SLE) susceptibility. METHODS: We collected all the publications about the relationship between CTLA-4 gene polymorphism and SLE susceptibility by searching PubMed, Web of Knowledge, Embase, Wanfang, CNKI and CBM before the date of May 20, 2012. The data were analyzed by the method of meta-analysis. RESULTS: A total of 12 studies were included, involving 3 gene locus, -1722, -1661 and -318. The results of total populations showed that there was a significant association between gene polymorphism and SLE risk in -1722 gene locus under a dominant model (OR=2.570, 95%CI=1.845-3.581, P<0.01) and -318 gene locus under a recessive model (OR=0.044, 95%CI=0.020-0.094, P<0.01). When stratified by population, the association still existed in Asian population, but no significant correlation existed in the other two populations (European and African). CONCLUSION: This meta-analysis supports that the CTLA-4 1722T/C and -318T/C polymorphisms are associated with SLE susceptibility, especially in Asian population.

[Role of TNF-alpha in vascular endothelial cells injury mediated by frozen/thawed PMN].

AIM: To investigate the role of TNF-alpha in vascular endothelial cells injury mediated by freezing/thaw ing PMN. METHODS: Freezing/thawing cell model was founded using rat PMN isolated by dextran sedimentation technique and VEC cultured in vitro. The injury level of VEC was indicated by measuring activity of LDH in medium. The number of frozen/thawed PMN adhering to VEC was counted with Phagocytizing reactive dyes the degree of frozen/thawed PMN and VEC adhesion. Expression of LFA-1 on the surface of frozen/thawed PMN was analyzed with flow cytometry. RESULTS: TNF-alpha could obviously upregulate expression of LFA-1 on surfaced of frozen/thawed PMN. Upregulation of LFA-1 expression promoted adhesion of frozen/thawed PMN and normal VEC,and aggravated VEC injury. Monoclonal antibody against LFA-1 could partly block adhesion of frozen/thawed PMN and normal VEC,and attenuate VEC injury. CONCLUSION: TNF-alpha can promote expression of LFA-1 on surface of frozen/thawed PMN adhering of frozen/thawed PMN to normal VEC and VEC injury increase, monoclonal antibody against LFA-1 could partly block PMN-VEC adhesion and attenuate VEC injury.