Effect of electroacupuncture at "Neiguan" (PC6) on pain and brain orexin 1 receptor in mice with inflammatory pain. / 电针"å† å ³"å¯¹ç‚Žæ€§ç—›å°é¼ è„‘å† é£Ÿæ¬²ç´ 1受体的影响.

OBJECTIVES: To observe the effect of electroacupuncture (EA) at "Neiguan" (PC6) on pain response in mice injected with complete Freund's adjuvant (CFA) in the hind paw, so as to investigate the mechanism of orexin 1 receptor (OX1R) -endogenous cannabinoid 1 receptor (CB1R) pathway in acupuncture analgesia. METHODS: A total of 48 male C57BL/6 mice were used in the present study. In the first part of this study, 18 mice were randomized into control, model and EA groups, with 6 mice in each group. In the second part of this study, 30 mice were randomized into control, model, EA, EA+Naloxone, EA+OX1R antagonist (SB33486) groups, with 6 mice in each group. Inflammatory pain model was established by subcutaneous injection of 20 µL CFA solution in the left hind paw. EA (2 Hz, 2 mA ) was applied to bilateral PC6 for 20 min, once a day for 5 consecutive days. The mice in the EA+Naloxone and EA+SB33486 groups were intraperitoneally injected with naloxone (10 mg/kg) or SB33486 (15 mg/kg) 15 min before EA intervention on day 5, respectively. Tail-flick method and Von Frey method were used to detect the thermal pain threshold and mechanical pain threshold of mice. Quantitative real-time PCR was used to detect the expression level of ß-endorphin mRNA in periaqueductal gray (PAG) of mice. The expression of OX1R positive cells in the lateral hypothalamic area (LH) and CB1R positive cells in the ventrolateral periaqueductal gray (vlPAG) were detected by immunofluorescence. RESULTS: Compared with the control group, the thermal pain threshold and mechanical pain threshold of the model group were decreased (P<0.001), the expression level of ß-endorphin mRNA in PAG was decreased (P<0.001), and the numbers of OX1R positive cells in LH and CB1R positive cells in vlPAG were decreased (P<0.05, P<0.001). Compared with the model group, the thermal pain threshold and mechanical pain threshold of the EA group were significantly increased (P<0.001), and the numbers of OX1R positive cells in LH and CB1R positive cells in vlPAG were increased (P<0.01, P<0.001). Compared with the EA group, the mechanical pain threshold in the EA+SB33486 group was significantly decreased (P<0.01), but there was no significant difference in the mechanical pain threshold between the EA+Naloxone group and EA group, and the numbers of OX1R positive neurons in LH and CB1R positive neurons in vlPAG were decreased in the EA+SB33486 group (P<0.001). CONCLUSIONS: EA at PC6 can achieve analgesic effect on CFA mice by activating the OX1R-CB1R pathway in the brain, and this effect is opioid-independent.

Effect of electroacupuncture on inflammatory response and immune cells in mice with chronic inflammatory pain. / ç”µé’ˆå¯¹æ ¢æ€§ç‚Žæ€§ç—›å°é¼ ç‚Žæ€§ååº”å’Œå ç–«ç»†èƒžçš„å½±å“.

OBJECTIVES: To observe the effects of electroacupuncture(EA) on local inflammatory mediators and macrophage polarization, and immune cells in the spleen of mice with chronic inflammatory pain induced by complete Freund's adjuvant (CFA) in the hind paw, so as to investigate the immunoinflammatory regulatory mechanisms of EA in relieving pain and swelling in mice with chronic inflammatory pain. METHODS: Thirty C57BL/6 mice were randomly divided into control, model, and EA groups, with 10 mice in each group. Chronic inflammatory pain model were established by subcutaneous injection of 20 µL CFA solution in the left hind paw for 7 consecutive days. After modeling, mice in the EA group received EA at bilateral "Zusanli"(ST36) for 20 min (2 Hz/100 Hz, 1 mA) once a day for 18 consecutive days. Mechanical pain threshold, heat pain thresholds, and paw thickness were measured before and after mode-ling, and after interventions. Western blot was used to detect the expression of tumor necrosis factor-alpha (TNF-α), interleukin (IL)-1ß, and NOD-like receptor protein 3 (NLRP3) in the paw tissue. Immunohistochemistry was used to detect the positive expression of M1-type macrophage marker inducible nitric oride synthase (iNOS) and M2-type marker CD206 in the paw, and flow cytometry was used to detect the proportion of F4/80+ CD11b+ macrophages, Ly6G+ CD11b+ neutrophils, and CD25+ Foxp3+ regulatory T cells (Treg) in the spleen. RESULTS: Compared with the control group, mechanical pain and heat pain thresholds were significantly reduced(P<0.000 1), while paw thickness, expressions of IL-1ß, TNF-α, and NLRP3 in the paw, and positive expression of M1 macrophage marker iNOS in the paw, the proportions of macrophages and neutrophils in the spleen were significantly increased (P<0.000 1, P<0.001) in the model group. Compared with the model group, mechanical pain threshold and heat pain thresholds, CD206 positive expression in the paw, and Treg cell proportion in spleen were significantly increased (P<0.01), while paw thickness, the expressions of IL-1ß, TNF-α and NLRP3 in the paw, as well as the positive expression of M1 macrophage marker iNOS in the paw, the proportions of macrophages and neutrophils in the spleen were significantly reduced (P<0.001, P<0.01, P<0.05)in mice of the EA group after intervention. CONCLUSIONS: EA may alleviate pain and swelling in mice with chronic inflammatory pain by regulating the numbers of macrophages, neutrophils, and Treg cells, as well as promoting M2 polarization of local macrophages and inhibiting the release of pro-inflammatory cytokines.

[Evaluation of the lower urinary tract function using voiding spot assay in mice].

The objective of present study was to develop a simple and reliable voiding spot assay (VSA) system to evaluate the lower urinary tract function of mice, and to establish it as a standardized protocol. Ultraviolet (UV) light was used to screen out the filter paper without autofluorescence and with optimal urine diffusion properties. Next, the appropriate wavelength of UV was determined based on the quality of the photographic image of urine spots on the filter paper. To confirm that the urine stain area on the filter paper was correlated with the amount of urine, a volume-area standard curve was constructed. The utility of this VSA system was validated using female wild-type C57BL/6J mice aged 12-13 weeks, and the data generated under identical procedural settings were compared among laboratories. Furthermore, this VSA system was employed to analyze the changes in voiding patterns in mice with urinary tract infections or transportation stress. No. 4 filter paper with a thickness of 0.7 mm was identified as the most suitable material for VSA, exhibiting no autofluorescence and facilitating optimal urine diffusion. The filter paper retained its integrity during the assay, and there was a linear correlation between urine volume and stained area under 365 nm UV light. Utilizing this VSA system, we determined that female wild-type C57BL/6J mice produced approximately 695.8 µL total urine and 5.5 primary voiding spots (PVS) with an average size of 126.4 µL/spot within 4-h period. Over 84% of PVS volumes ranged from 20 to 200 µL. Notably, PVS volumes of mice were similar across different laboratories. Mice with urinary tract infections or transportation stress exhibited significant changes in VSA parameters, including increased voiding frequency, PVS number, and decreased PVS volume. Therefore, this VSA system can be used to evaluate the urinary function of normal mice, as well as those with urinary tract infection or transportation stress.

Hypoxic Tumor-Derived Exosomal miR-199a-3p Promote Gastric Cancer Metastasis via MAP3K4.

Proximal gastrectomy is more frequently recommended for early upper gastric cancer and Siewert II gastroesophageal junction cancer less than 4 cm in length. After proximal gastrectomy, the anatomical structure of the gastroesophageal junction can be destroyed, and the anti-reflux effect of the cardia is lost. In recent years, as various anti-reflux reconstructions have been developed, some functions of the stomach are retained, and serious reflux esophagitis is avoided after proximal gastrectomy. In this article, we summarized the indications, advantages, and disadvantages of various classic reconstruction and latest improved reconstruction method including esophageal and residual stomach anastomosis, tubular gastroesophageal anastomosis, muscle flap anastomosis, jejunal interposition, and double-tract reconstruction.

[Effect of different electrical current intensities of electroacupuncture preconditioning on cardiac function and macrophage polarization in mice with acute myocardial ischemia].

OBJECTIVE: To observe the effect of different intensities of electroacupuncture (EA) preconditioning on car-diac function and polarization state of macrophages in mice with acute myocardial ischemia (AMI), so as to explore its possible mechanism underlying improvement of AMI. METHODS: A total of 50 male C57BL/6J mice were randomly divided into sham ope-ration, AMI model, and EA pretreatment groups (0.5 mA, 1 mA, 3 mA subgroups), with 10 mice in each group/subgroup. The mice in the EA pretreatment groups were subjected to EA stimulation of bilateral "Neiguan"(PC6) with 0.5, 1.0 and 3 mA respectively and frequency of 2 Hz/15 Hz for 20 min, once a day, for 3 days. The acute myocardial ischemia model was established by ligating the anterior descending branch (ADB) of the left coronary artery, while the sham operation only had a surgical suture trans-passed below the ADB but without ligation. The myocardial infarction area was measured after TTC staining, and the cardiac function ï¼»left ventricular ejection fraction (EF), short-axis contraction rate (FS)ï¼½ was detected by using echocardiography. The M1 macrophages were labeled with CD11b+F480+CD206low, M2 macrophages were labeled with CD11b+F480+CD206high and detected by using flow cytometry, and the expression levels of myocardial interleukin-1ß (IL-1ß), tumor necrosis factor-α (TNF-α), Toll-like receptor-4 (TLR4) proteins were detected by using Western blot. RESULTS: Compared with the sham operation group, the model group had a significant increase in the infarction area (P<0.000 1), number of cardiac macrophages and percentage of M1 type macrophages (P<0.000 1), and the expression levels of myocardial IL-1ß, TNF-α, TLR4 proteins (P<0.001, P<0.01), and a remarkable decrease in the levels of EF, FS and the percentage of M2 type macrophages (P<0.000 1). In contrast to those of the model group, the area of myocardial infarction (P<0.000 1, P<0.01), expression levels of myocardial IL-1ß, TNF-α, TLR4 proteins (P<0.01, P<0.05, P<0.001) in the 0.5 mA, 1 mA and 3 mA groups, number of macrophages and percentage of M1 macrophages (P<0.05) in the 1 mA group were significantly decreased, while the levels of EF and FS (P<0.000 1, P<0.05, P<0.001) in the 3 EA groups, and percentage of M2 macrophage (P<0.05) in the 1 mA group were significantly increased. Comparison among the 3 EA groups displayed that the effects of 1 mA group were significantly superior to those of 0.5 and 3 mA groups in up-regulating EF and FS (P<0.01, P<0.001), and in down-regulating the area of infarct myocardium (P<0.01, P<0.000 1), and the expression of TLR4 protein (P<0.01), and 0.5 mA group in the expression of IL-1ß and TNF-α proteins (P<0.05). CONCLUSION: EA preconditioning with electrical current intensities of 0.5 mA, 1 mA and 3 mA can effectively reduce myocardial infarction size, improve cardiac function in mice with AMI, which may be related with its effects in reducing the number of cardiac macrophages and down-regulating the expression of myocardial IL-1ß, TNF-α and TLR4 proteins. The therapeutic effect of 1 mA is better than that of 0.5 and 3 mA.

Research progress of L-aspartate-α-decarboxylase and its isoenzyme in the ß-alanine synthesis.

Driven by the massive demand in recent years, the production of ß-alanine has significantly progressed in chemical and biological ways. Although the chemical method is relatively mature compared to biological synthesis, its high cost of waste disposal and environmental pollution does not meet the environmental protection standard. Hence, the biological method has become more prevalent as a potential alternative to the chemical synthesis of ß-alanine in recent years. As a result, the aspartate pathway from L-aspartate to ß-alanine (the most significant rate-limiting step in the ß-alanine synthesis) catalyzed by L-aspartate-α-decarboxylase (ADC) has become a research hotspot in recent years. Therefore, it is vital to comprehensively understand the different enzymes that possess a similar catalytic ability to ADC. This review will investigate the exploratory process of unique synthesis features and catalytic properties of ADC/ADC-like enzymes in particular creatures with similar catalytic capacity or high sequence homology. At the same time, we will discuss the different ß-alanine production methods which can apply to future industrialization.

Sugammadex for reversing neuromuscular blockages after lung surgery: A systematic review and meta-analysis.

BACKGROUND: This study determined whether sugammadex was associated with a lower risk of postoperative pulmonary complications and improved outcomes in lung surgeries. METHODS: A systematic literature search was conducted using PubMed, Embase, Web of Science, and the Cochrane Library from January 2000 to March 2022. The characteristics of lung surgeries using sugammadex treatment compared with control drugs and postoperative outcomes were retrieved. The primary outcome was estimated through a pooled odds ratio (OR) and its 95% confidence interval (CI) was identified using a random-effects model. RESULTS: From 465 citations, 7 studies with 453 patients receiving sugammadex and 452 patients receiving a control were included. The risk of postoperative pulmonary complication (PPCs) was lower in the sugammadex group than in the control group. Also, it showed that the effect of sugammadex on PPCs in the subgroup analysis was significantly assessed on the basis of atelectasis or non-atelectasis. Furthermore, subgroup analysis based on the relationship between high body mass index (BMI) and PPCs also showed that sugammadex had less occurrence in both the high BMI (defined as BMI ≥ 25) and low BMI groups. No difference in length of hospital stay (LOS) between the two groups was observed. CONCLUSION: This study observed that although reversing neuromuscular blockages with sugammadex in patients undergoing thoracic surgery recorded fewer PPCs and shorter extubation periods than conventional reversal agents, no difference in LOS, postanaesthesia care unit (PACU) stay length and chest tube insertion duration in both groups was observed.

Predictors of unfavorable outcome at 90 days in basilar artery occlusion patients.

BACKGROUND: In a previous study, basilar artery occlusion (BAO) was shown to lead to death or disability in 80% of the patients. The treatment for BAO patients in the acute stage includes thrombolysis and intra-arterial thrombectomy, but not all patients benefit from these treatments. Thus, understanding the predictors of outcome before initiating these treatments is of special interest. AIM: To determine the predictors related to the 90-d clinical outcome in patients with BAO in an Asian population. METHODS: We performed a retrospective case review of patients admitted to a tertiary stroke center between 2015 and 2019. We used the international classification of diseases-10 criteria to identify cases of posterior circulation stroke. A neurologist reviewed every case, and patients fulfilling the criteria defined in the Basilar Artery International Cooperation Study were included. We then analyzed the patients' characteristics and factors related to the 90-d outcome. RESULTS: We identified a total of 99 patients as real BAO cases. Of these patients, 33 (33.3%) had a favorable outcome at 90 d (modified Rankin Scale: 0-3). Moreover, 72 patients received intra-arterial thrombectomy, while 13 patients received intravenous tissue-type plasminogen activator treatment. We observed a favorable outcome in 33.3% of the cases and an unfavorable outcome in 66.7% of the cases. We found that the initial National Institutes of Health Stroke Scale (NIHSS) score and several BAO symptoms, including impaired consciousness, tetraparesis, and pupillary abnormalities, were significantly associated with an unfavorable outcome (P < 0.05), while cerebellar symptoms were associated with a favorable outcome (P < 0.05). In the receiver operating characteristic (ROC) analysis, the areas under the ROC curve of initial NIHSS score, impaired consciousness, tetraparesis, cerebellar symptoms, and pupillary abnormalities were 0.836, 0.644, 0.727, 0.614, and 0.614, respectively. Initial NIHSS score showed a higher AUROC (0.836) compared to BAO symptoms. CONCLUSION: The most important predictor of an unfavorable outcome was the initial NIHSS score. BAO symptoms, including tetraparesis, impaired consciousness, and pupillary abnormality were also related to an unfavorable outcome.

[Transfusion Strategy for Rare Anti-LW Antibody Autoimmune Hemolytic Anemia Patients with Neonatal Hemolytic Disease].

OBJECTIVE: To analyze the causes of positive irregular antibody screening test and incompatibility of cross matching in one patient with autoimmune hemolytic anemia complicated with neonatal hemolytic disease, and to accurately identify the type of antibodies in patients, and to select a reasonable strategy for blood transfusion. METHODS: One children was enrolled, blood group positive and reverse typing, Rh typing, direct anti-human globulin test, free test, dispersal test and cross matching test were carried out by test tube method and microcolumn gel card; irregular antibodies were identified by the reaction of DTT treatment and untreated panel cells with patients' plasma. RESULTS: The blood group of the patient was RhD positive B and irregular antibody screening positive, while the blood group of the mother was RhD positive O and irregular anti-screening negative, the result showed that the anti-LW detected in the plasma of the patient was autoantibody and ABO neonatal hemolytic disease (ABO-HDN) was present. Both O type RhD positive washing RBCs and B type RhD negative RBCs were transfused effectively. CONCLUSION: Irregular antibodies in patients are anti-LW antibodies, and transfusion of homotype RhD negative suspended erythrocytes after the exclusion of ABO-HDN shows a better effect.

NMDAR in bladder smooth muscle is not a pharmacotherapy target for overactive bladder in mice.

Overactive bladder (OAB) is a common condition that affects a significant patient population. The N-methyl-D-aspartate receptor (NMDAR) has a role in developing bladder overactivity, pharmacological inhibition of which inhibits bladder overactivity. The common pathogenesis of OAB involves bladder smooth muscle (BSM) overactivity. In this study, a smooth muscle-specific NMDAR knockout (SMNRKO) mouse model was generated. The bladders from SMNRKO mice displayed normal size and weight with an intact bladder wall and well-arranged BSM bundles. Besides, SMNRKO mice had normal voiding patterns and urodynamics and BSM contractility, indicating that NMDAR in BSM was not essential for normal physiological bladder morphology and function. Unexpectedly, cyclophosphamide (CYP)-treated SMNRKO and wild-type (WT) mice had similar pathological changes in the bladder. Furthermore, SMNRKO mice displayed similar altered voiding patterns and urodynamic abnormalities and impaired BSM contractility compared with WT mice after CYP treatment. MK801 partially reversed the pathological bladder morphology and improved bladder dysfunction induced by CYP, but did not cause apparent differences between WT mice and SMNRKO mice, suggesting that NMDAR in BSM was not involved in pathological bladder morphology and function. Moreover, the direct instillation of NMDAR agonists or antagonists into the CYP-induced OAB did not affect bladder urodynamic function, indicating that NMDAR in BSM was not the pharmacotherapy target of MK801 for CYP-induced cystitis. The findings indicated that NMDAR in BSM was not essential for normal physiological or pathological bladder morphology and function, and MK801 improving pathological bladder function was not mediated by an action on NMDAR in BSM.

[Distribution of Redox Zone at Different Water Layers in the Presence of Periphyton and the Responsible Microorganisms].

So far, many types of carriers (such as artificial mat, industrial soft carriers) have been widely used in removing pollutants, purifying water quality via the periphyton attached on the surface of these carriers. In the presence of periphyton, the distribution of redox zone at different water layers is directly or indirectly associated with the removal rate of pollutants. Therefore, it is more practically significant to study the distribution of redox zone at different water layers and the microbial diversity in the presence of periphyton. In this study, the pilot experiment was performed in a simulated water column bioreactor. Firstly, the eutrophic water collected from XuanWu Lake was added into the simulated water column bioreactor. The industrial soft carriers were then suspended into the water column in order to enhance the growth of periphyton. After periphyton gained a steady growth state, the oxidation reduction zones (redox zones) and the responsible microorganisms at different water layers were monitored. The results showed that five sequent redox zones (i. e. oxygen reduction, nitrate reduction, iron reduction, methanogenic and sulfate reduction zones, respectively) appeared in different water layers from top-down in the presence of periphyton and their responsible terminal electron acceptors were O2, NO3(-), Fe3+, CO2 and SO4(2-) respectively. The indicators of the different zones were DO, NO2(-), Fe(2+), HCO3(-) and sulfide, and the highest concentrations were 11.290 mg x L(-1), 4.950 mg x L(-1), 38.326 mg x L(-1), 120.000 mg x L(-1) and 12.180 mg x L(-1), respectively. The results of microbiological characteristics tested by Biolog EcoPlate technology revealed that there were significant differences in the composition, metabolic activity, carbon utilization of periphyton at different water layers, causing the difference in the distribution of redox zones at different water layers. These findings implies that study on the distribution of redox zones and microbiological characteristics in the presence of periphyton provides a better understanding that periphyton is capable of improving water quality at different layer, and also provides some theoretical basis for the development of technology for purifying water quality based on periphyton.

[Characteristics of sludge in treatment of high concentrated organic wastewater by swim-bed technology].

Based on the technology of conditional activated sludge process, swim-bed technology involving the novel biomass attachment material biofringe (BF) is one of the new and efficient advanced municipal wastewater treatments process. The process demonstrated effective treatment of high concentrated organic wastewater. When VLRs were from 1.7 kg/(m3 x d) to 4.5 kg/(m3 x d), 96% of average COD removal efficiencies and 80.7% of nitrification rate were achieved, respectively. The BF material allowed for attachment of large amounts of biomass. The mixed liquor suspended solid (MLSS) concentrations in the reactor reached 23 g/L at the highest during operation period, with holding 20-50 mL/g of SVI. An exponential law was found to be suitable to describe the variation of the viscosity as a function of MLSS. The granulation phenomenon of sludge was observed during the experiment. Protozoan and metazoans on the bio-film and suspended sludge were proved existing in the system by microscopic observation and DNA experiment. Observed sludge yields from 0.1046 to 0.1723 (MLSS/BOD) were obtained in swim-bed system.