Electroacupuncture improves allodynia and central sensitization via modulation of microglial activation associated P2X4R and inflammation in a rat model of migraine.

Background: Recent studies have demonstrated that activated microglia were involved in the pathogenesis of central sensitization characterized by cutaneous allodynia in migraine. Activation of microglia is accompanied by increased expression of its receptors and release of inflammatory mediators. Acupuncture and its developed electroacupuncture (EA) have been recommended as an alternative therapy for migraine and are widely used for relieving migraine-associated pain. However, it remains rare studies that show whether EA exerts anti-migraine effects via inhibiting microglial activation related to a release of microglial receptors and the inflammatory pathway. Therefore, this study aimed to investigate EA' ability to ameliorate central sensitization via modulation of microglial activation, microglial receptor, and inflammatory response using a rat model of migraine induced by repeated epidural chemical stimulation. Methods: In the present study, a rat model of migraine was established by epidural repeated inflammatory soup (IS) stimulation and treated with EA at Fengchi (GB20) and Yanglingquan (GB34) and acupuncture at sham-acupoints. Pain hypersensitivity was further determined by measuring the mechanical withdrawal threshold using the von-Frey filament. The changes in c-Fos and ionized calcium binding adaptor molecule 1 (Ibal-1) labeled microglia in the trigeminal nucleus caudalis (TNC) were examined by immunflurescence to assess the central sensitization and whether accompanied with microglia activation. In addition, the expression of Ibal-1, microglial purinoceptor P2X4, and its associated inflammatory signaling pathway mediators, including interleukin (IL)-1ß, NOD-like receptor protein 3 (NLRP3), and Caspase-1 in the TNC were investigated by western blot and real-time polymerase chain reaction analysis. Results: Allodynia increased of c-Fos, and activated microglia were observed after repeated IS stimulation. EA alleviated the decrease in mechanical withdrawal thresholds, reduced the activation of c-Fos and microglia labeled with Ibal-1, downregulated the level of microglial purinoceptor P2X4, and limited the inflammatory response (NLRP3/Caspase-1/IL-1ß signaling pathway) in the TNC of migraine rat model. Conclusions: Our results indicate that the anti-hyperalgesia effects of EA ameliorate central sensitization in IS-induced migraine by regulating microglial activation related to P2X4R and NLRP3/IL-1ß inflammatory pathway.

Acupoint Selection in Postoperative Ophthalmic Pain Management: A Data Mining Protocol.

Background: Postoperative ophthalmic pain not only induces anxiety and depression among patients, but also prolongs the recovery cycle. However, the management of postoperative pain in ophthalmology is still not standardized. The effectiveness of acupuncture in treating postoperative pain has been validated based on numerous clinical trials and meta-analysis. Our study is to conduct the first data mining analysis to identify the most effective acupoints selection and combinations for treating postoperative ophthalmic pain, inform. Methods: We will search bibliographic databases from inception to November 2023. Clinical trials evaluating the effectiveness of acupuncture therapy in the management of postoperative ophthalmic pain will be selected. Reviews, protocols, animal studies, case reports, systematic evaluations and Meta-analyses will be excluded. Primary outcome indicators will be clinical outcomes related to postoperative ophthalmic pain. Descriptive statistics will be performed in Excel 2019. Association rule analysis will be performed in SPSS Modeler 18.0. Exploratory factor analysis and cluster analysis will be performed in SPSS Statistics 25.0. Results: This study will investigate the most effective point selection and combination of acupuncture points for the treatment of postoperative ophthalmic pain. Conclusion: Our findings will provide evidence for the effectiveness and potential therapeutic prescription of acupuncture for postoperative ophthalmologic pain, helping clinicians and patients work together to make more informed decisions.

Analysis of Traditional Chinese Medicine Symptoms in Children with Spastic Cerebral Palsy: A Data Mining Study.

Background: Cerebral palsy (CP) ranks as a major cause of motor disabilities in children, with spastic CP making up roughly 70-80% of all CP cases. The primary objective of our study is to identify characteristics of Traditional Chinese Medicine(TCM) symptom of spastic CP, thereby establishing correlations between the TCM symptom and the disease, providing a more scientific theoretical foundation for TCM treatments on spastic CP, enabling a deeper comprehension of clinical interventions, ultimately, improving rehabilitation outcomes in TCM treatment for spastic CP. Methods: We conducted a data mining study on TCM symptom of spastic CP children aged 4-14 years old treated at Xi'an Encephalopathy Hospital Affiliated to Shaanxi University of Chinese Medicine, from October 2021 to March 2023. The medical records of all eligible and complete spastic CP patients were extracted, processed for data cleansing, transformed, and subsequently analyzed to discern distinctive TCM symptom. K-Means Clustering Analysis and Association Rule Analysis were used for data mining. Results: Core symptoms identified for spastic CP encompassed "Motor Dysfunction", "Impaired Speech", "Delayed Development", "Limb Stiffness", "Rigidity in the limbs", "Intellectual Impairment", "Timidity and susceptibility to startle responses", "Muscle Wasting", and "Pale or Dull Complexion". Among the top-ranking associations of symptom, patterns emerge wherein "Motor dysfunction" intertwine with "Impaired speech", "Motor dysfunction" coexist with "Delayed development", and "Impaired speech" are accompanied by "Delayed development". Conclusion: This study identified the core symptom of spastic CP and tentatively suggests that the clinical manifestations of spastic CP are essentially consistent with the TCM pattern "liver exuberance and spleen weakness". This finding has facilitated the preliminary establishment of correlations between TCM pattern differentiation and the disease in medicine. It is anticipated that this correlation will bring tangible benefits to a larger number of children with spastic CP.

Electroacupuncture Reduces Inflammatory Bowel Disease in Obese Mice by Activating the Nrf2/HO-1 Signaling Pathways and Repairing the Intestinal Barrier.

Background: Electroacupuncture (EA) is used to treat inflammatory bowel disease (IBD). Nevertheless, the precise mechanisms by which this approach safeguards against obesity-induced intestinal barrier damage has not been fully understood. Objective: This study aimed to assess whether EA could ameliorate intestinal barrier damage that had been reversed in a mouse model of obesity induced by a high-fat diet (HFD) and whether this repair is correlated with ferroptosis and gut microbiota enhancement. Methods: To assess the potential of EA to prevent obesity and restore the intestinal barrier, we divided in C57BL/6J mice into two groups; one was fed with HFD and another one with a normal diet. Samples of stool, blood, fat, and intestinal epithelium were then evaluated, along with body weight. Results: Following EA, we observed a significant reduction in body weight, fat accumulation, and serum triglyceride (TG), total cholesterol (TC), and low-density lipoprotein cholesterol (LDL-C) levels; an increase was seen in high-density lipoprotein cholesterol (HDL-C) levels. EA also activated the Nrf2 signaling pathway; upregulated the expression of GPX4, FTH1, and SLC7A11; and downregulated the expression of TFR1. In addition, the administration of EA resulted in a notable modification of the gut microbiota composition, characterized by a decrease in the Firmicutes to Bacteroidetes ratio. Conclusion: EA had beneficial effects on weight loss and showed potential ability to repair the intestinal barrier by activating the Nrf2 signaling pathway, inhibiting intestinal inflammation and ferroptosis, and regulating the intestinal microbiota to treat IBD caused by HFD-induced obesity.

Antibiotic Skeletal Diversification via Differential Enoylreductase Recruitment and Module Iteration in trans-Acyltransferase Polyketide Synthases.

Microorganisms are remarkable chemists capable of assembling complex molecular architectures that penetrate cells and bind biomolecular targets with exquisite selectivity. Consequently, microbial natural products have wide-ranging applications in medicine and agriculture. How the "blind watchmaker" of evolution creates skeletal diversity is a key question in natural products research. Comparative analysis of biosynthetic pathways to structurally related metabolites is an insightful approach to addressing this. Here, we report comparative biosynthetic investigations of gladiolin, a polyketide antibiotic from Burkholderia gladioli with promising activity against multidrug-resistant Mycobacterium tuberculosis, and etnangien, a structurally related antibiotic produced by Sorangium cellulosum. Although these metabolites have very similar macrolide cores, their C21 side chains differ significantly in both length and degree of saturation. Surprisingly, the trans-acyltransferase polyketide synthases (PKSs) that assemble these antibiotics are almost identical, raising intriguing questions about mechanisms underlying structural diversification in this important class of biosynthetic assembly line. In vitro reconstitution of key biosynthetic transformations using simplified substrate analogues, combined with gene deletion and complementation experiments, enabled us to elucidate the origin of all the structural differences in the C21 side chains of gladiolin and etnangien. The more saturated gladiolin side chain arises from a cis-acting enoylreductase (ER) domain in module 1 and in trans recruitment of a standalone ER to module 5 of the PKS. Remarkably, module 5 of the gladiolin PKS is intrinsically iterative in the absence of the standalone ER, accounting for the longer side chain in etnangien. These findings have important implications for biosynthetic engineering approaches to the creation of novel polyketide skeletons.

Analysis of Traditional Chinese Medicine Symptoms in Children with Spastic Cerebral Palsy, a Protocol for Data Mining.

Background: Cerebral palsy (CP) is characterized by abnormal pronunciation, posture, and movement. Clinically, CP can be categorized into various motor syndromes, including spastic hemiplegia, diplegia, quadriplegia, involuntary movement, ataxia, and mixed types. Among these, spastic CP represents over 70-80% of all CP cases. The primary objective of our study is to identify the top and core Traditional Chinese Medicine (TCM) symptoms and analysis their association rules in children with spastic cerebral palsy, thereby enhancing the theoretical foundations of TCM treatment on spastic CP. Methods: The study will be conducted on children aged 4 to 14 years with spastic CP who are undergoing treatment at Xi'an Encephalopathy Hospital Affiliated to Shaanxi University of Chinese Medicine. Basic information about the patients and their TCM symptoms will be collected on the first day of admission. This information will include age, gender, birth history, family history, disease classification, and TCM symptoms (including symptoms, tongue, and pulse). Once the data is collected, it will be exported from the electronic medical record system for further analysis. Descriptive statistics will be performed using Excel 2019, while exploratory factor analysis and cluster analysis will be conducted using SPSS Statistics 22. Additionally, association rule analysis will be carried out using SPSS Modeler 18. Results: This study will investigate the most top TCM symptoms in children with spastic CP and explore the association rules between these symptoms, mapping the presentation of spastic CP onto symptoms identified within TCM. Conclusion: Our findings will provide the distinctive characteristics of TCM symptoms in children with spastic CP, furnishing evidence-based support to clinicians and patients in making well-informed decisions collaboratively.

Analysis on the Selection of Herbs in TCM COVID-19 Treatment Protocols Between Malaysia and China.

Objective: Both Malaysian and Chinese populations have benefited from the utilization of Traditional Chinese Medicine (TCM) in the treatment of COVID-19. This study aims to analyze the herb selection in TCM COVID-19 treatment protocols employed by Malaysia and China. Methods: Based on TCM theories, a comparative analysis was conducted on the treatment protocols of the two countries. This analysis encompassed syndrome differentiation and classification, therapeutic principles and formulas, as well as herb characteristics. The herbs within the protocols were subjected to frequency statistics and cluster analysis using SPSS 25, and network analysis was performed using SPSS Modeler 18.0 software. Results: The properties (cold, warm, ping and bitter, pungent, sweet) and meridian (lung, stomach, spleen) associated with the herbs in both protocols exhibit fundamental similarities. Furthermore, the propensity for selecting pairs of herbs is also consistent. The herb pairs selected also have the same propensity. The clustering results reveal six categories, of which in the two protocols are good. The top three herb-pairs in Malaysian Protocols are Gan Cao - Yi Yi Ren, Ban Xia - Gan Cao, Chen Pi - Gan Cao, while the top three in Chinese Protocol are Huo Xiang - Ma Huang, Shi Gao - Gan Cao, Cang Zhu - Gan Cao. Conclusion: Characteristics of herbs in Both Malaysian and Chinese Protocols herb reflect the advantages and regional adaptation of TCM principles. These findings offer valuable guidance in the application of herbs or herb pairs for the treatment of COVID-19.

Electroacupuncture at Fengchi(GB20) and Yanglingquan(GB34) Ameliorates Paralgesia through Microglia-Mediated Neuroinflammation in a Rat Model of Migraine.

BACKGROUND: Multiple studies have suggested that paralgesia (hyperalgesia and cutaneous allodynia) in migraine reflects the activation and sensitisation of the trigeminovascular system (TGVS). In particular, it reflects the second-order and higher nerve centre sensitisation, which is caused and maintained by neuroinflammation. Microglia activation leads to the release of proinflammatory cytokines involved in inflammatory responses. Accumulating evidence indicates that electroacupuncture (EA) is effective in ameliorating paralgesia, but the underlying mechanisms of EA in migraine attacks caused by microglia and microglia-mediated inflammatory responses are still unclear. The purpose of this study was to explore whether EA could ameliorate the dysregulation of pain sensation by suppressing microglial activation and the resulting neuroinflammatory response, and to evaluate whether this response was regulated by Toll-like receptor 4 (TLR4)/nuclear factor-kappa B(NF-κB) in the trigeminal nucleus caudalis (TNC) in a rat model of migraine. METHODS: Repeated Inflammatory Soup (IS) was infused into the dura for seven sessions to establish a recurrent migraine-like rat model, and EA treatment was administered at Fengchi (GB20) and Yanglingquan (GB34) after daily IS infusion. Facial mechanical withdrawal thresholds were measured to evaluate the change in pain perception, and plasma samples and the TNC tissues of rats were collected to examine the changes in calcitonin gene-related peptide (CGRP), the Ibal-1-labelled microglial activation, and the resulting inflammatory response, including interleukin-1ß (IL-1ß), tumour necrosis factor-α (TNF-α), interleukin-6 (IL-6), and their regulatory molecules TLR4/NF-κB, via enzyme-linked immunosorbent assay (ELISA), real-time polymerase chain reaction (RT-PCR), immunohistochemistry (IHC) and Western blot analysis. RESULTS: Repeated IS injections into the dura induced facial mechanical paralgesia, which is the manifestation of migraine attacks, and increased the expression of CGRP, Ibal-1, microglial mediated inflammatory cytokines (IL-1ß, TNF-α, IL-6), and regulatory molecules TLR4/NF-κB. EA at GB20/34 significantly attenuated repetitive IS-induced pain hypersensitivity. This effect was consistent with decreased levels of CGRP and inflammatory cytokines in the plasma and the TNC via the inhibition of microglia activation, and this response may be regulated by TLR4/NF-κB. CONCLUSIONS: EA ameliorated paralgesia in repetitive IS-induced migraine-like rats, which was mainly mediated by a reduction in microglial activation and microglial-mediated inflammatory responses that could be regulated by TLR4/NF-κB.

Electroacupuncture Alleviates Depressive-like Behavior by Modulating the Expression of P2X7/NLRP3/IL-1ß of Prefrontal Cortex and Liver in Rats Exposed to Chronic Unpredictable Mild Stress.

Depression is a complex clinical disorder associated with poor outcomes. Electroacupuncture (EA) has been demonstrated to have an important role in both clinical and pre-clinical depression investigations. Evidence has suggested that the P2X7 receptor (P2X7R), NLRP3, and IL-1ß play an important role in depressive disorder. Our study is aimed at exploring the role of EA in alleviating depression-like behaviors in rats. We therefore investigated the effects of EA on the prefrontal cortex and liver of rats subjected to chronic unpredictable mild stress (CUMS) through behavior tests, transmission electron microscopy, Nissl staining, HE staining, immunohistochemistry and Western blotting. Five weeks after exposure to CUMS, Sprague-Dawley (SD) rats showed depression-like behavior. Three weeks after treatment with brilliant blue G (BBG) or EA, depressive symptoms were significantly improved. Liver cells and microglia showed regular morphology and orderly arrangement in the BBG and EA groups compared with the CUMS group. Here we show that EA downregulated P2X7R/NLRP3/IL-1ß expression and relieved depression-like behavior. In summary, our findings demonstrated the efficacy of EA in alleviating depression-like behaviors induced by CUMS in rats. This suggests that EA may serve as an adjunctive therapy in clinical practice, and that P2X7R may be a promising target for EA intervention on the liver-brain axis in treatment of depression.

Upregulation of OTX2-AS1 is Associated With Immune Infiltration and Predicts Prognosis of Gastric Cancer.

BACKGROUND: It is unclear whether the long non-coding RNA (lncRNA) OTX2 antisense RNA 1 (OTX2-AS1) plays a pivotal role in gastric cancer (GC). An analysis of The Cancer Genome Atlas (TCGA) database data and bioinformatics was used to explore the relationship between OTX2-AS1 and GC in the current study. METHODS: We evaluated the relationship between clinical features and OTX2-AS1 expression, prognostic factors, and the significant involvement of OTX2-AS1 in function using various statistical methods, such as Kaplan-Meier method, Cox regression analysis, Gene Set Enrichment Analysis (GSEA), and immune infiltration analysis. GC cell lines were tested for OTX2-AS1 expression using qRT-PCR. RESULTS: A high level of OTX2-AS1 expression was significantly and negatively associated with Helicobacter pylori (H pylori) infection in GC patients (P = .006) and predicted a poorer overall survival (OS) (HR: 1.54; 95% CI: 1.10-2.14; P = .011), progression-free interval (PFI) (HR: 1.75; 95% CI: 1.22-2.51; P = .002) and disease-specific survival (DSS) (HR: 1.85; 95% CI: 1.21-2.85; P = .005) in GC patients. There was an independent correlation between OTX2-AS1 expression (HR: 1.771; 95% CI: 1.164-2.696; P = .008) and OS in patients with GC. There were differential enrichments for the OTX2-AS1 high expression phenotype in the olfactory transduction, G alpha (s) signaling events, keratinization, olfactory signaling pathway, and preimplantation embryo. OTX2-AS1 expression may be related to certain immune-infiltrating cells. Compared to gastric epithelial cells (GES-1), GC cell lines showed a significant increase in OTX2-AS1 expression. CONCLUSION: There was a significant association between OTX2-AS1 expression in GC patients and poor survival, suggesting that it may be a useful biomarker for prognosis and immunotherapy outcome of stomach adenocarcinoma (STAD) in GC.

[Effect of electroacupuncture on cognitive impairment in APP/PS1 mice based on TLR4/NF-κB/NLRP3 pathway].

OBJECTIVE: To observe the effect of electroacupuncture (EA) on the expressions of tight junction related proteins Claudin-5, ZO-1 in the colon and hippocampus, Toll-like receptor 4/nuclear factor-kappa B/NOD-like receptor protein 3 (TLR4/NF-κB/NLRP3) pathway in the hippocampus of APP/PS1 mice, so as to explore its mechanisms underlying improvement of cognitive impairment. METHODS: Eighteen 5-month-old male APP/PS1 mice were equally randomized into model and EA groups,and nine 5-month-old male C57BL/6 mice were used as the normal control. EA(2 Hz, 1 mA) was applied to "Baihui" (GV20), "Dachangshu" (BL25) and "Zusanli" (ST36) for 15 min, once daily, 5 days a week for 5 weeks. The Morris water maze swimming test was used to evaluate the mice's cognitive impairment. Nissl staining was used to observe the pathological morphology of hippocampus. The expression of amyloid ß-peptide (Aß) in brain tissue was detect by immunohistochemistry; the contents of lipopolysaccharide (LPS) in colon, serum and hippocampus were detected by ELISA; the expression levels of Claudin-5, ZO-1 in colon and hippocampus, and TLR4/NF-κB/NLRP3 pathway related proteins in hippocampus were detected by Western blot. RESULTS: Compared with the normal group, the escape latency of the mice in the model group was prolonged from the 3rd day (P<0.05, P<0.01), the number of crossing the platform and the percentage of target quadrant residence time were significantly decreased (P<0.01), and the contents of LPS in colon, serum and hippocampus were significantly increased (P<0.01), the expression levels of TLR4, NF-κB p65, NLRP3, Caspase-1, interleukin (IL)-1ß and tumor necrosis factor (TNF)-α in hippocampus and Aß in brain tissue were significantly increased (P<0.01), while the expression levels of Claudin-5, ZO-1 in colon and hippocampus were significantly decreased (P<0.01). Compared with the model group, the escape latency of mice in the EA group was shortened from the 4th day (P<0.05, P<0.01), the number of crossing the platform and the percentage of target quadrant residence time were increased (P<0.01, P<0.05), and the contents of LPS in serum and hippocampus were decreased (P<0.05), and the expression levels of TLR4, NF-κB p65, Caspase-1, NLRP3, IL-1ß, TNF-α in hippocampus and Aß in brain tissue were significantly decreased (P<0.05, P<0.01), while the expression levels of Claudin-5, ZO-1 in colon and hippocampus were significantly increased (P<0.05, P<0.01). Outcomes of Nissl staining showed dispersed arrangement of neurons with nuclear pyknosis or hyperchromasia in the hippocampus, and a decreased number of cell layers in the model group, which was relatively milder in the EA group. CONCLUSION: EA may improve the cognitive impairment of APP/PS1 mice by up-regulating the expression of Claudin-5 and ZO-1, reducing the transposition of gut-derived LPS to the central nervous system, inhibiting the over-activation of TLR4/NF-κB/NLRP3 pathway, and alleviating the inflammatory reaction of the central nervous system.

[Effect of mild moxibustion on vascular aging in senescence rats based on local MCP-1/MMP-2/TGF-ß1 pro-inflammatory signal loop].

OBJECTIVE: To observe the effect of mild moxibustion on monocyte chemotaxis protein 1 (MCP-1)/matrix metalloproteinase 2 (MMP-2)/transforming growth factor ß1 (TGF-ß1) pro-inflammatory signal loop in senile rats, so as to explore its mechanisms underlying improving vascular aging (VA). METHODS: Twenty-four male VA SD rats were randomized into senium (VA) control, medication and moxibustion groups, and other 8 young SD rats (aged 2 months) were used as the young control group. The VA model was established by intraperitoneal injection of D-galactose (300 mg·kg-1·d-1) once daily for 4 weeks, and verified by serum total testosterone (TT) and free testosterone (FT) levels. For rats of the moxibustion group, mild moxibustion was applied to bilateral "Shenshu"(BL23) and "Guanyuan"(CV4) for 20 min, once a day, 5 days a week for 8 weeks. Rats of the medication group were treated by intraperitoneal injection of testosterone propionate (7 mg·kg-1·[3 d]-1) once daily for 8 weeks except weekends, and rats of the senium control and young control groups treated by intraperitoneal injection of the same dose of normal saline, once daily for 8 weeks except weekends. The duration of exhausted swimming (DES) before and after the treatment was recorded. H.E. staining and Masson staining were used to observe histopathological changes and collagen fiber content of the thoracic aortic tissue, respectively. The contents of serum TT, FT and angiotensin 2 (Ang Ⅱ) were determined by ELISA. The immunoactivity of aortic MCP-1 was detected by immunohistochemistry, and the expression levels of aortic MCP-1, MMP-2 and TGF-ß1 were detected by Western blot. RESULTS: Compared with the young control group, the levels of DES, serum TT and FT contents were significantly decreased (P<0.01), while those of serum AngⅡ and collagen fiber contents, aortic MCP-1 immunoactivity and MMP-2, TGF-ß1 and MCP-1 protein expression considerably increased in the senium control group (P<0.01). After the interventions, the decreased levels of DES, serum TT and FT contents and the increased levels of serum AngⅡ, collagen fiber contents, aortic MCP-1 immunoactivity and MMP-2, TGF-ß1 and MCP-1 protein expression were reversed in both medication and moxibustion groups (P<0.01, P<0.05). The therapeutic effect of mild moxibustion was significantly superior to that of medication in down-regulating the aortic collagen fiber, serum AngⅡ contents and MCP-1 immunoactivity and protein expression (P<0.05). H.E. staining showed thickened endometrium and disordered arrangement of vascular smooth muscles of the aorta in the senium group, and thinner endometrium and regular and ordered arrangement of aortic vascular smooth muscles in both moxibustion and medication groups. CONCLUSION: Mild moxibustion may improve vascular aging in senescence rats, which is possibly by suppressing vascular MCP-1/MMP-2/TGF-ß1 pro-inflammatory signal loop.

[Effect of electroacupuncture on hepatocyte autophagy and oxidative stress in SAMP8 mice by regulating AMPK/mTOR/ULK1 signaling pathway].

OBJECTIVE: To observe the effect of electroacupuncture (EA) on physical strength and expression levels of hepatic AMP-activated protein kinase (AMPK), mammalian target of rapamycin (mTOR), unc-51 like autophagy activating kinase 1 (ULK1) proteins and Atg5, Atg7, Atg13, Beclin1 and ULK1 mRNAs in aging (senescence accelerated mouse/prone 8, SAMP8)mice, so as to exp lore its mechanism underlying delaying aging by activating AMPK/mTOR/ULK1 signaling pathway. METHODS: Twenty-four male SAMP8 mice were randomly divided into model group, rapamycin (autophagy inducer) group, EA group and EA+autophagy inhibitor (EA+inhibitor) group, with 6 mice in each group, and 6 homologous anti-rapid aging male (SAMR1) mice in the same age were used as the control group. Mice of the rapamycin group received intraperitoneal injection of rapamycin solution (2 mg·kg-1·d-1). EA (2 Hz, 1 mA) was applied to bilateral "Taichong"(LR3)and "Shenshu"(BL23) for 15 min each time. Mice of the EA+inhibitor group received intraperitoneal injection of mTOR inhibitor 3-methyladenine (1.5 mg·kg-1·d-1) before the EA intervention each time. The above-mentioned interventions were conducted 6 times a week for 2 consecutive weeks. Physical conditions of mice were assessed by exhaustive swimming tests. Histopathological changes of the liver were observed by H.E. staining. Western blot was used to detect the expression of AMPK, phosphorylated AMPK (p-AMPK), mTOR, phosphorylated mTOR (p-mTOR), ULK1 and phosphorylated ULK1 (p-ULk1) in the liver tissues. The expression levels of Atg5, Atg7, Atg13, Beclin1 and ULK1 (cellular autophagy-related genes) mRNAs in the liver were detected by quantitative real-time PCR. The immunoactivity (IA) of heme oxygenase 1 (HO-1) in the liver was detected by immunohistochemistry, and the activity of superoxide dismutase (SOD) and the content of malondialdehyde (MDA) of the liver were measured by hydroxylamine method for assessing the level of oxidative stress. RESULTS: Compared with the control group, the duration of exhaustive swimming, the expression levels of AMPK, p-AMPK, ULK1 and p-ULK1 proteins, and Atg5, Atg7, Atg13, Beclin1 and ULK1 mRNA, HO-1 IA and SOD activity were considerably down-regulated (P<0.01), while the expression levels of mTOR and p-mTOR and MDA content were significantly up-regulated (P<0.01) in the model group. In comparison with the model group, the duration of the exhausted swimming, the expression levels of AMPK, p-AMPK, ULK1 and p-ULK1 proteins, and Atg5, Atg7, Atg13, Beclin1 and ULK1 mRNAs, HO-1 IA and SOD activity were significantly up-regulated (P<0.01, P<0.05), whereas the expression levels of mTOR and p-mTOR proteins and MDA content were notably down-regulated (P<0.01, P<0.05) in the rapamycin, EA and EA+inhibitor groups. The improvement of the abovementioned indexes of EA+inhibitor group was not as good as rapamycin and EA groups (P<0.01), suggesting an elimination of the therapeutic effects after administration of 3-methyladenine. No significant differences were found between the rapamycin and EA groups in the abovementioned indexes (P>0.05) except p-mTOR and mTOR which were higher in the EA group (P<0.01). H.E. staining showed ambiguous boundary of the liver lobule, disordered arrangement of hepatocytes with a large amount of fat vacuoles at different size and deviation of nucleus, and lysis of some hepatocytes. These situations were relatively milder in the rapamycin and EA groups. CONCLUSION: EA may enhance physical strength and promote cellular autophagy in the liver of aging mice by regulating AMPK/mTOR/ULK1 signaling, thereby inhibiting excessive oxidative stress, and delaying aging process to some extent.

[Effects of electroacupuncture on autophagy factors and liver lipid metabolism in rapidly aging mice].

Objective: To study the effects of electroacupuncture on the expressions of autophagy-related factors LC3-Ⅱ, Beclin1, Atg7, and P62 in the liver of rapidly aging (senescence accelerated mouse/prone8,SAMP8) mice, and to explore the mechanisms of electroacupuncture to improve liver lipid metabolism in mice. Methods: Thirty-week-old male SAMP8 mice were randomly divided into model group, drug group, and electroacupuncture group, with 7 mice in each group. Seven anti-rapid aging SAMR1 mice of the same age were used as the control group. The animals in the control group and the model group were bred routinely for 2 weeks without any intervention; the drug group was treated with intraperitoneal injection of rapamycin at the dose of 10 mg·kg-1·d-1, once a day, 6 consecutive days a week; the electroacupuncture group was given "Shenshu" and "Taichong" Electroacupuncture at point(15 minutes a day, 6 consecutive days a week). The serum lipid metabolism and liver lipid deposition of mice were detected, the distribution of liver autophagy body, the protein and mRNA expressions of liver LC3 - Ⅱ, Beclin1, Atg7 and P62 were determined. Results: Compared with the control group, the total cholesterol (TC), triglycerides (TG), and low density lipoprotein (LDL) of the model group were increased significantly(P＜0.01). In the model group, lipid droplet deposition was obvious, autophagosomes were decreased, the protein and mRNA expression levels of autophagy- related factors LC3-Ⅱ, Beclin1 and Atg7 were decreased significantly (P＜0.01), while the protein and mRNA expressions of P62 were increased significantly (P＜0.01). Compared with the model group, the serum contents of TG, TC, and LDL of the mice in the electroacupuncture group and the drug group were decreased significantly (P＜0.01), lipid droplet deposition was reduced, autophagosomes were increased, the protein and mRNA expression levels of LC3 -Ⅱ, Beclin1 and Atg7 were increased significantly(P＜0.01), and the protein and mRNA expression levels of P62 were decreased significantly(P＜0.01). The protein and mRNA expression levels of Beclin1 and Atg7 in the liver of the electroacupuncture group were not significantly different from the drug group (P＞0.05). Conclusion: Electroacupuncture can alleviate liver lipid metabolism disorders, which may be related to the regulation of the expressions of liver autophagy related factors LC3-Ⅱ, Beclin1, Atg7, and P62, thereby promoting liver autophagy in SAMP8 mice.

The Prevalence of Pulmonary Hypertension Among Maintenance Dialysis Patients With ESRD and Its Associated Factors: A Retrospective Study.

Aim: To determine the prevalence of pulmonary hypertension (PH) and its associated factors among end-stage renal disease (ESRD) patients who underwent maintenance dialysis. Methods: A total of 491 patients received echocardiography examinations and underwent pulmonary artery systolic pressure (PASP) assessments. A subgroup of 283 patients were subjected to plasma creatinine (Cr) and blood urea nitrogen concentration (BUN) tests, routine blood examinations and electrolyte analysis. First, we compared the differences in echocardiographic, Cr and BUN, blood routine and electrolyte parameters between PH and non-PH groups. The correlations between PASP and the parameters mentioned above were also analyzed. Furthermore, univariate and adjusted logistic regression analyses were performed to identify the independent associated factors. Results: The incidence of PH among ESRD patients who were treated with maintenance dialysis was 34.6%. Most of the echocardiographic parameters, including end-diastolic internal diameters of the left atrium, left ventricle, right atrium, and pulmonary artery, as well as interventricular septum mobility, left ventricular posterior wall mobility, fractional shortening, stroke volume and left ventricle ejection fraction (LVEF), were associated with PH. Furthermore, Mg2+ (p = 0.037) and Cl- (p = 0.043) were significantly associated with PASP. However, after adjustments were made in the regression analysis, only internal diameters of the left atrium, right atrium, and LVEF were independently associated with PH. Conclusion: PH is prevalent, with a relatively high incidence among ESRD patients who undergo maintenance dialysis. The sizes of the left and right atria as well as LVEF were independently associated with PH, but further cohort and basic mechanistic studies are needed to confirm this finding.

The expression of epidermal growth factor receptor 1 and human epidermal growth factor receptor 2 based on tumor location affect survival in gastric cancer.

To investigate the different expression of epidermal growth factor receptor 1 (EGFR) and human epidermal growth factor receptor 2 (HER2) in gastric cancer based on tumor locations and its impact on patients survival.Gastric cancer is heterogeneous disease, recent years have established a molecular classification and described distribution of molecular subtypes in stomach. However, the difference of EGFR and HER-2 expression among tumor location is still unknown.Between January 2010 and August 2014, 2477 consecutive patients with gastric cancer were treated in our surgery department. The tumor locations were classified into 4 groups: cardia, fundus, corpus, and antrum. Based on tumor locations, the clinicopathologic characteristics, EGFR and HER-2 expression, and follow-up data were analyzed by univariant analysis and Kaplan-Meier analysis retrospectively.There were difference of gender, age, Borrmann type, pathological type, differentiation, T-stage, tumor size, gastrectomy method, and complications among the locations. The positive rate of EGFR expression in fundus was 18.18%, which was lower than cardia (46.21%), corpus (43.62%), and antrum (48.83%) (P < .001). The 5-year survival rate in EGFR positive patients was 50.8%, which was significantly lower than EGFR negative patients (64.0%, P = .021). The positive rate of HER-2 expression in cardia was 48.15%, which was significantly higher than fundus (37.5%), corpus (35.45%), and antrum (38.54%) (P = .009), but HER-2 expression did not correlate with 5-year survive (P = .548).Our results suggest that there exist difference of EGFR and HER-2 expression based on tumor locations, and the distribution of EGFR impact on patients survival. Emphasizing the role of EGFR and HER-2 in the context of location contribute to make appropriate treatment strategy and improve prognosis of gastric cancer.

Convalescent Plasma in the Treatment of Severe COVID-19: A Systematic Review and Meta-Analysis.

BACKGROUND: COVID-19 is a public health emergency of international concern. Its incidence rates and mortality are very high; however, so far, an effective drug treatment remains unknown. Based on the role of convalescent plasma therapy in previously identified viral pneumonias, patients with severe COVID-19 have been given this therapy. This systematic review and meta-analysis aimed to summarize the clinical evidence regarding the efficacy and safety of convalescent plasma therapy in the treatment of severe COVID-19. METHODS: PubMed, Embase, Ovid, China Knowledge Network, China Biomedical, VIP Chinese Sci-tech Journal, Wanfang Database, and the International Clinical Trials Registry Platform were searched up to 21 June 2020, to identify clinical studies and registered trials on the use of convalescent plasma in the treatment of critically ill patients with COVID-19. Stata 13.0 was used to perform Meta-analysis. All records were screened as per the protocol eligibility criteria. RESULTS: Nineteen clinical reports regarding convalescent plasma in the treatment of severe COVID-19 were included. Through systematic analysis, convalescent plasma was found to yield some efficacy on severe COVID-19 and had almost no obvious adverse reactions. CONCLUSION: Convalescent plasma therapy seems to yield some efficacy among patients with severe COVID-19 and almost no obvious adverse reactions were found. However, at present, the clinical evidence is insufficient, and there is an urgent need for support from high-quality clinical trial data.

[Effects of electroacupuncture on proangiogenesis process and protein turnover in a mouse model of sarcopenia].

OBJECTIVE: To observe the effect of electroacupuncture(EA) on proangiogenesis process and protein turn-over in a mouse model of sarcopenia, so as to explore its potential molecular mechanism anti-aging. METHODS: Fourteen 30-week-old male SAMP8 mice were randomly divided into a model group (n=7) and an EA group (n=7). Seven anti-rapidly aging SAMR1 mice of the same age were used as the control group (n=7). EA (1 mA, 4 Hz) was applied to bilateral "Zusanli"(ST36) and "Yanglingquan"(GB34) for 20 minutes each time once a day, 6 times a week for 4 weeks. The exhausted running platform was used to test the sports function. Gastrocnemius muscle mass and relative ratio of gastrocnemius muscle mass to body mass were measured. HE staining and transmission electron microscope were used to observe the morphology, and the cross-sectional area of gastrocnemius muscle was calculated. Relative protein expressions of protein kinase B (AKT) , phosphorylated (p) -AKT, mammalian target of rapamycin (mTOR) , p-mTOR, p70 ribosomal protein S6 kinase (p70S6K) , p-p70S6K,hypoxia inducible factor-1α (HIF-1α) and relative mRNA expressions of HIF-1α, vascular endothelial growth factor A (VEGF-A) , muscle RING finger-1 (MuRF-1) and muscle atrophy F-box (MAFbx) were detected by Western blot and real-time fluorescence quantitative PCR, seperatively. RESULTS: Compared with the control group, the running time and distance, body mass and gastrocnemius mass, and the ratio of gastrocnemius mass to body mass decreased(P<0.01, P<0.05), cross-sectional area of gastrocnemius, related protein expression of p-AKT,p-mTOR, p-p70S6K and HIF-1α, mRNA expression of HIF-1α and VEGF-A decreased (P<0.01), while mRNA expression of MuRF1 and MAFbx increased (P<0.01) in the model group. Following EA intervention, the running time and distance, body mass and gastrocnemius mass and the ratio of gastrocnemius mass to body mass increased (P<0.05), cross-sectional area of gastrocnemius, related protein expression of p-AKT,p-mTOR, p-p70S6K and HIF-1α, mRNA expression of HIF-1α and VEGF-A were significantly up-regulated (P<0.01), mRNA expression of MuRF1 and MAFbx down-regulated (P<0.01, P<0.05) in the EA group compared with the model group. CONCLUSION: EA may delay the aging muscle atrophy in mice by regulating the gastrocnemius muscle's proangiogenesis process and protein turnover.

The Prevalence and Independent Risk Factors of Significant Tricuspid Regurgitation Jets in Maintenance Hemodialysis Patients With ESRD.

BACKGROUND AND AIM: Tricuspid regurgitation (TR) is a frequent complication in various cardiovascular diseases. However, few studies have reported the prevalence of TR especially the moderate to severe or significant TR (ms-TR) maintenance dialysis patients. Thus, we aimed to identify the prevalence of ms-TR and its associated factors. METHODS: A total of 491 maintenance dialysis patients underwent echocardiographic examinations, while a subgroup (n = 283) also received routine blood tests, renal function examinations, and electrolyte analysis. We first compared the differences in abovementioned parameters among groups with various TR areas (TRAs). Finally, univariate and adjusted regression were also used to identify factors that were independently associated with ms-TR. RESULTS: The incidence of TR jets was 62.6%, which included a mildly increased TRA (47.8%), moderately increased TRA (10.4%), and severely increased TRA (3.5%). Most of the cardiac structures and functional parameters, such as the end-diastolic internal diameters of the left atrium (LA), left ventricle (LVDD), right atrium (RA), right ventricle (RV), left ventricular ejection fraction (LVEF), and fractional shortening (FS), were significantly associated with ms-TR. Among serum ions, only total CO2 (TCO2; r = -0.141, p = 0.047) was negatively correlated with TRA. After adjusted, only Na+ [odds ratio (OR): 0.871 0.888, p = 0.048], RA (OR: 1.370, p < 0.001), and FS (OR: 0.887, p < 0.001) were independently associated with ms-TR. CONCLUSION: Tricuspid regurgitation occurs in maintenance hemodialysis patients with ESRD. Na+ FS and RA were independently associated with ms-TR, and these parameters may be potential risk factors/predictors for ms-TR.

ß-MnO2 /Metal-Organic Framework Derived Nanoporous ZnMn2 O4 Nanorods as Lithium-Ion Battery Anodes with Superior Lithium-Storage Performance.

Nanoporous ZnMn2 O4 nanorods have been successfully synthesized by calcining ß-MnO2 /ZIF-8 precursors (ZIF-8 is a type of metal-organic framework). If measured as an anode material for lithium-ion batteries, the ZnMn2 O4 nanorods exhibit an initial discharge capacity of 1792 mA h g-1 at 200 mA g-1 , and an excellent reversible capacity of 1399.8 mA h g-1 after 150 cycles (78.1 % retention of the initial discharge capacity). Even at 1000 mA g-1 , the reversible capacity is still as high as 998.7 mA h g-1 after 300 cycles. The remarkable lithium-storage performance is attributed to the one-dimensional nanoporous structure. The nanoporous architecture not only allows more lithium ions to be stored, which provides additional interfacial lithium-storage capacity, but also buffers the volume changes, to a certain degree, during the Li+ insertion/extraction process. The results demonstrate that nanoporous ZnMn2 O4 nanorods with superior lithium-storage performance have the potential to be candidates for commercial anode materials in lithium-ion batteries.