Inhibitory effect of acupoint electrostimulation with different layers and intensities on muscular inflammatory pain and spinal WDR neuron activity. / ç©´ä½ä¸åŒå±‚æ¬¡å’Œå¼ºåº¦ç”µåˆºæ¿€ç¼“è§£è‚Œè‚‰ç‚Žæ€§ç—›åŠå¯¹è„Šé«“èƒŒè§’å¹¿åŠ¨åŠ›ç¥žç»å ƒçš„æŠ‘åˆ¶ä½œç”¨.

OBJECTIVES: To observe the analgesic effects of different levels and intensities of electrical stimulation on the local acupoints in the pain source area and their impact on wide dynamic range (WDR) neurons in the spinal dorsal horn, in order to provide a basis for selecting appropriate parameters for electroacupuncture (EA) stimulation. METHODS: Wistar rats were used in 3 parts of the experiment. Complete Freund's adjuvant was used to establish a model of inflammation-induced pain in the gastrocnemius muscle. After modeling, 6 rats were randomly selected for multi-channel extracellular electrophysiological recording of the electrical activity of WDR neurons, to determine the threshold for activating the A-component (Ta) and the C-component (Tc), which were used as the intervention intensities for skin transcutaneous electrical acupoint stimulation (TEAS) or EA. Thirty-six rats were randomly divided into normal , model , TEAS-Ta , TEAS-Tc, EA-Ta , and EA-Tc groups, with 6 rats in each group. In the pain source area , Ta or Tc intensity of TEAS or EA intervention at"Chengshan"(BL57) was performed for 30 min each time, once a day, for 3 consecutive days. A small animal pressure pain measurement instrument was used to measure the mechanical pressure pain threshold of the gastrocnemius muscle in rats, and the Von Frey filament was used to measure the mechanical pain threshold of the footpad. Thirteen rats were randomly selected to observe the immediate responsiveness of WDR neurons to Ta/Tc intensity of EA or TEAS in BL57. RESULTS: The thresholds of TEAS to activate WDR neuron A-component or C-component were (2.43±0.57) mA and (7.00±1.34) mA, respectively, while the thresholds for EA to activate muscle WDR neuron A-component or C-component were (0.72±0.34) mA and (1.58±0.35) mA, respectively. After injection of CFA into the gastrocnemius muscle, compared with the normal group both the mechanical pressure pain threshold of the gastrocnemius muscle and the mechanical pain threshold of the footpad of rats in the model group were significantly decreased (P<0.001). After TEAS-Ta, TEAS-Tc or EA-Ta intervention in the BL57, both the mechanical pressure pain threshold of the gastrocnemius muscle and the mechanical pain threshold of the footpad were significantly higher than those in the model group (P<0.05, P<0.001). Compared with the normal group, the electrical threshold for evoking WDR neuron C-component discharge was significantly decreased (P<0.001) in the model group, while increased after TEAS-Ta, TEAS-Tc, or EA-Ta intervention (P<0.01) compared with the model group. The evoked discharge frequency of muscle WDR neurons decreased significantly after immediate intervention with TEAS-Ta, TEAS-Tc, or EA-Ta (P<0.01, P<0.05). EA-Tc had no significant improvement on the evoked electrical activity of WDR neurons or pain behavior. CONCLUSIONS: TEAS-Ta, TEAS-Tc, or EA-Ta can all alleviate the local and footpad mechanical pain in rats with muscle inflammation and inhibit the responsiveness of WDR neurons, indicating that different intensities are required for analgesic effects at different levels of acupoints in the pain source area.

Local analgesia of electroacupuncture is mediated by the recruitment of neutrophils and released ß-endorphins.

ABSTRACT: The efficacy of acupuncture in treating pain diseases has been recognized in clinical practice, and its mechanism of action has been a hot topic in academic acupuncture research. Previous basic research on acupuncture analgesia has focused mostly on the nervous system, with few studies addressing the immune system as a potential pathway of acupuncture analgesia. In this study, we investigated the effect of electroacupuncture (EA) on the ß-endorphins (ß-END) content, END-containing leukocyte type and number, sympathetic neurotransmitter norepinephrine (NE), and chemokine gene expression in inflamed tissues. To induce inflammatory pain, about 200 µL of complete Frester adjuvant (CFA) was injected into the unilateral medial femoral muscle of adult Wistar rats. Electroacupuncture treatment was performed for 3 days beginning on day 4 after CFA injection, with parameters of 2/100 Hz, 2 mA, and 30 minutes per treatment. The weight-bearing experiment and enzyme-linked immunosorbent assay showed that EA treatment significantly relieved spontaneous pain-like behaviors and increased the level of ß-END in inflamed tissue. Injection of anti-END antibody in inflamed tissue blocked this analgesic effect. Flow cytometry and immunofluorescence staining revealed that the EA-induced increase in ß-END was derived from opioid-containing ICAM-1 + /CD11b + immune cells in inflamed tissue. In addition, EA treatment increased the NE content and expression of ß2 adrenergic receptor (ADR-ß2) in inflammatory tissues and upregulated Cxcl1 and Cxcl6 gene expression levels. These findings provide new evidence for the peripheral analgesic effect of acupuncture treatment by recruiting ß-END-containing ICAM-1 + /CD11b + immune cells and increasing the ß-END content at the site of inflammation.

The Effect of Pre-Electroacupuncture on Nociceptive Discharges of Spinal Wide Dynamic Range Neurons in Rat.

Purpose: Spinal wide dynamic range (WDR) neurons are well studied in pain models and they play critical roles in regulating nociception. Evidence has started to accumulate that acupuncture produces a good analgesic effect via activating different primary fibers with distinct intensities. The purpose of the present study was to compare the distinct intensities of pre-electroacupuncture (pre-EA) at local muscular receptive fields (RFs), adjacent or contralateral non-RFs regulating the nociceptive discharges of spinal WDR neurons evoked by hypertonic saline (HS). Materials and Methods: Spinal segments of electrophysiological recording were identified by neural tracers applied at the left gastrocnemius muscle. The thresholds of Aß (TAß), Aδ (TAδ) and C (TC) components of WDR neurons were measured to determine the intensity of pre-EA by extracellular recording. The discharges of WDR neurons induced by distinct intensities of pre-EA and 200 µL HS (6%) injection in left gastrocnemius muscle of rats were observed by extracellular recording. Results: The spinal segments of WDR neurons were confirmed in lumbar (L)5-6 area according to the projective segments of dorsal root ganglion. TAß, TAδ and TC of WDR neurons was determined to be 0.5, 1, and 2 mA, respectively. The pre-EA with intensities of TAß (P < 0.05), TAδ (P < 0.05), TC (P < 0.05) or 2TC (P < 0.01) at ipsilateral adjacent non-RFs significantly reduced the discharges of WDR neurons, while at local RFs only pre-EA of TAδ (P < 0.05), TC (P < 0.05) and 2TC (P < 0.01) could inhibit the nociceptive discharges. In addition, intensity of pre-EA at contralateral non-RFs should reach at least TC to effectively inhibit the firing rates of WDR neurons (P < 0.01). Conclusion: Pre-EA could suppress nociceptive discharges of WDR neurons and the inhibitory effects were dependent on the distinct intensities and locations of stimulation.

Damage characteristics of fabrics created by TASER probes.

TASER® weapons are conducted energy weapons (CEWs) that are frequently used by police departments around the world. CEWs can be deployed in two methods: drive stun application and probe deployment. This study aims to examine damages caused by TASER devices on fabrics and whether types of fabric material and TASER models could contribute to different damage features. Three types of white fabric were used, including 100% cotton, 100% polyester, and 65:35 polyester-cotton blend. Three models: TASER X26P, TASER X2, and TASER 7 were shot onto each type of fabric, with five repetitions each. Each damaged area on the fabric caused by a probe is a sample (n = 90) and was examined with a Keyence digital microscope. Images were captured by the Keyence microscope and measurements were recorded, including damage dimensions, fabric condition, evidence of burning, and extra findings. The presence of fused yarn ends was found to be statistically significant across the fabric types, and no damage features were found that may assist in the identification of TASER models. Other damage features including damage dimensions, discoloration, and fiber deformation were not found to be showing apparent differences according to statistical analysis. The conclusions made by this research should be used with caution due to the small sample size.

TASER CEW distance determination for models X26P, X2, and TASER 7.

TASER weapons, classified as Conducted Energy Weapons (CEWs) are used by law enforcement as electrical, non-lethal weapons. A CEW's long distance method of deployment launches two barbed probes that embed into the target, before electricity is cycled from one probe to the other. The resulting muscle contractions help incapacitate the intended target. Currently, there is no research on the accuracy of variability of CEW probe spread in determining the distance of the shooter. Thus, in this study, a method of distance determination for TASER models X26P, X2, and TASER 7 was established to help estimate the relative locations of individuals during crime scene reconstruction. Each of four probe cartridge types were launched for six repetitions towards vertical cardboard targets from three different distances (n = 72). The probe spread on the targets was measured for each cartridge and plotted against CEW-To-Target (CTT) distance. Linear regression trendlines and standard deviations were generated for each cartridge type. Results demonstrated probe spread distances to be proportional to probe launch angles, and greater CTT distances resulted in greater standard deviation. Standard error of the estimate was calculated for all four linear regression equations and their ability to predict CTT distance when given probe spread measurements. The mean standard error values appeared acceptable for the X2 (0.460 m), TASER 7 CQ (0.273 m), and X26P (0.322 m) CEW models, while the mean error calculated for the TASER 7 Standoff model (0.846 m) was considerably greater. We expect our findings will help establish a reliable method of distance determination for CEWs, and perhaps with a larger sample size, more thoroughly examine the accuracy of the linear regression equations. CEW distance determination has the potential to provide support in contextualizing crime scene reconstruction with further research. By providing distance estimations alongside a standardized guide of error allowance, it would be possible to induce how far apart individuals were at the time a CEW was fired, within the calculated window of deviation, and in a statistical degree of confidence.

Manifestation of TASER drive stun burn marks on fabrics.

The TASER® is a type of conducted energy weapon (CEW) used with increasing frequency by law enforcement to subdue subjects in circumstances where compliance is necessary. When operated in the drive stun method of deployment, the electrodes at the head of this CEW are intended to make direct contact with a surface, generating heat and light which may result in burn marks as a by-product of the electrical discharge that occurs. This research aims to tackle a crucial gap in CEW research that fails to address the appearance of burn marks on fabrics. A drive stun duration (DSD) of 1, 3, and 5 s was used with three TASER models (X26P, X2, & TASER 7) on three fabrics (white 100% cotton, 100% polyester, 35:65 cotton-polyester blend) with an underlying backing of pork hock. Using a Keyence VHX-6000 confocal microscope, high magnification images were taken to observe any qualitative changes to the fabric. On polyester fabric, with increasing DSD, darker brown discoloration occurred. Additionally, on polyester fabric, the spatial orientation of the burn marks corresponded with that of the electrodes at the muzzle of each TASER model. These features enabled the correct identification of the TASER model and DSD on polyester fabric in the blind tests performed. Evidence of burn marks on cotton and blend fabrics were both limited and inconsistent such that no features were sufficiently unique to link them to any TASER model or DSD. Despite this, overall features such as discoloration and the formation of holes on these fabric types may be sufficiently unique to suggest the use of a CEW.

Noninvasive nasopharyngeal proteomics of COVID-19 patient identify abnormalities related to complement and coagulation cascade and mucosal immune system.

Serum or plasma have been the primary focus of proteomics studies for COVID-19 to identity biomarkers and potential drug targets. The nasal mucosal environment which consists of lipids, mucosal immune cells, and nasal proteome, has been largely neglected but later revealed to have critical role combating SARS-CoV-2 infection. We present a bottom-up proteomics investigation of the host response to SARS-CoV-2 infection in the nasopharyngeal environment, featuring a noninvasive approach using proteins in nasopharyngeal swabs collected from groups of 76 SARS-CoV-2 positive and 76 negative patients. Results showed that 31 significantly down-regulated and 6 up-regulated proteins were identified (p < 0.05, log2 FC > 1.3) in SARS-CoV-2 positive patient samples as compared to the negatives; these proteins carry potential value as markers for the early detection of COVID-19, disease monitoring, as well as be drug targets. The down-regulation of coagulation factor 5 indicates a thrombotic abnormality in COVID-19 patients and the decreased IgG4 suggests an abnormal immune response at the point of entry in human nasopharyngeal environment, which is in consistent with KEGG and GO pathway analysis. Our study also demonstrated that mass spectrometry proteomics analysis of nasopharyngeal swabs can be used as a powerful early approach to evaluate host response to SARS-CoV-2 viral infection.

Demonstrating Hairy and Glabrous Skin Innervation in a 3D Pattern Using Multiple Fluorescent Staining and Tissue Clearing Approaches.

Skin innervation is an important part of the peripheral nervous system. Although the study of the cutaneous nerve fibers has progressed rapidly, most of the understanding of their distributional and chemical characteristics comes from conventional histochemical and immunohistochemical staining on thin tissue sections. With the development of the tissue clearing technique, it has become possible to view the cutaneous nerve fibers on thicker tissue sections. The present protocol describes multiple fluorescent staining on tissue sections at a thickness of 300 µm from the plantar and dorsal skin of rat hindfoot, the two typical hairy and glabrous skin sites. Here, the calcitonin gene-related peptide labels the sensory nerve fibers, while phalloidin and lymphatic vessel endothelial hyaluronan receptor 1 label the blood and lymphatic vessels, respectively. Under a confocal microscope, the labeled sensory nerve fibers were followed completely at a longer distance, running in bundles in the deep cutaneous layer and freestyle in the superficial layer. These nerve fibers ran in parallel to or surrounded the blood vessels, and lymphatic vessels formed a three-dimensional (3D) network in the hairy and glabrous skin. The current protocol provides a more effective approach to studying skin innervation than the existing conventional methods from the methodology perspective.

Synergistic effects of Rapamycin and Fluorouracil to treat a gastric tumor in a PTEN conditional deletion mouse model.

The tumor suppressor gene phosphatase and tensin homolog (PTEN) in PI3K/Akt/mTOR pathway is essential in inhibiting tumor growth and metastasis. However, whether the mutation of PTEN gene could induce tumorigenesis and impact the treatment of gastric cancer is still unclear. The purpose of the study was to investigate the combined treatment of gastric tumorigenesis using Rapamycin and Fluorouracil (5-Fu) through interfering with the Akt/mTOR pathway in a mouse model with PTEN conditional deletion. Three groups of mice were exposed for 5 days to Rapamycin and 5-Fu separately and together. The gene expression of the Akt/mTOR pathway, the protein expression of caspase-3 and p-Akt, p-S6K and p-4EBP1, and the pathological changes in stomachs were analyzed. Our study demonstrates that the conditional PTEN deletion in the cells of glandular stomach induces hyperplastic gastric tumors in mice. The combined Rapamycin administration with 5-Fu resulted in better outcomes than their separate administration for the treatment of gastric cancer by inhibiting the mTOR signal pathway. Our study indicates that Rapamycin has a synergistic interaction with chemotherapeutic 5-Fu, and demonstrates a potential therapeutic combination treatment on glandular stomach tumor with PTEN functional absence or aberrantly activated Akt/mTOR pathway. It provides important insights into the inhibition of the Akt/mTOR pathway in gastric cancer clinical therapy.

The Homeless People in China During the COVID-19 Pandemic: Victims of the Strict Pandemic Control Measures of the Government.

Background: By implementing aggressive control measures, China has rapidly and effectively controlled the Coronavirus disease 2019 (COVID-19) pandemic. However, the neglected homeless population may become victims of that perceived success. Due to political sensitivity, we know little about them. Aims: This study aimed to investigate how the pandemic and the pandemic control strategies of the government affected the lives of the homeless people in mainland China. Methods: A total of 103 eligible participants experiencing absolute homelessness were recruited from Guangzhou City during July and August 2020. Surveys measured demographic characteristics including health status, changes in daily living, and actions of the government toward the homeless during the pandemic. Sankey diagrams and ordered logit regression models were used to examine the impact on the homeless of inhumane government efforts to drive the homeless away. Qualitative materials were analyzed by using an inductive approach to provide more details. Results: First, the homeless people in Guangzhou tended to be male, aged 40 to 64 years, less well-educated, and they originated from outside Guangdong although they were living in the city center. Most had little connection with their families. After a long period of homelessness, almost half of the participants were in poor health with various conditions, which made them extremely vulnerable to COVID-19. Second, the pandemic caused a substantial decline in incomes of homeless people, had less of a negative impact on their food intake, and affected sleep time in different ways. Third, during the COVID-19 pandemic, humanitarian aid from local governments of China decreased, whereas inhumane efforts to drive the homeless away intensified. Fourth, quantitative models and qualitative materials demonstrate the devastating effect of the strict pandemic control strategy of the government on the lives of homeless people, which may further cause their health problems. Conclusion: This study for the first time illustrated the characteristic features of the street homeless population in mainland China and their living situation during the COVID-19 pandemic, and most importantly demonstrated the devastating effect of the strict pandemic control of the government, which has been considered a great success in previous studies, on lives of homeless people. Urgent measures should be taken to ensure the protection of the homeless population and prevent an impending humanitarian crisis.

Tracking Gluten throughout Brewing Using N-Terminal Labeling Mass Spectrometry.

Gluten-containing grains cause adverse health effects in individuals with celiac disease. Fermentation of these grains results in gluten-derived polypeptides with largely uncharacterized sizes and sequences, which may still trigger an immune response. This research used N-terminal labeling mass spectrometry to characterize protein hydrolysates during each stage of bench-scale brewing, including malting, mashing, boiling, fermentation, and aging. Gluten hydrolysates from each brewing step were tracked, and the immunotoxic potential was evaluated by sequence comparison with peptides known to stimulate celiac immune responses. The results indicate that proteolysis and precipitation of gliadins occurring during brewing differ by protein region and brewing stage. The termini of gliadins were hydrolyzed throughout the entire brewing process, but the central regions remained relatively stable. Most hydrolysis occurred during malting, and most precipitation occurred during boiling. The addition of yeast yielded new cleavage sites but did not result in complete hydrolysis. Consistent detection of peptides within the clinically important regions of gliadin corroborated the hydrolytic resistance of this region. N-terminal labeling mass spectrometry served as a novel approach to track the fate of gliadin/gluten throughout bench-scale brewing. Consistently identified fragments could serve as improved targets for the detection of hydrolyzed gluten in fermented products.

Compositional and immunogenic evaluation of fractionated wheat beers using mass spectrometry.

The safety and regulatory status of fermented products derived from gluten-containing grains for patients with celiac disease remains controversial. Bottom-up mass spectrometry (MS) has complemented immunoassays for the compositional and immunogenic analyses of wheat beers. However, uncharacterized proteolysis during brewing followed by the secondary digestion for MS has made the analysis and data interpretation complicated. In this study, the composition and immunogenic potential of seven commercially available wheat beers were evaluated using bottom-up MS with the aid of fractionation and a multi-step peptide search strategy to identify peptides generated by various types of proteolysis. Gluten-derived peptides accounted for approximately 50% and 20% of the total number of wheat-derived and barley-derived peptides, respectively, in the investigated beers. Although relatively large polypeptides cannot be thoroughly characterized using traditional bottom-up proteomics, up to 50% of peptides identified contained celiac-immunogenic motifs, and consumption of wheat beers would pose risks for celiac patients.

Evaluation of N-terminal labeling mass spectrometry for characterization of partially hydrolyzed gluten proteins.

Gluten, a group of proteins found in wheat, barley, and rye, is the trigger of celiac disease, an immune disorder that affects about 1% of people worldwide. The toxicity of partially hydrolyzed gluten (PHG) in fermented products is less well understood due to the significant analytical challenges in PHG characterization. In this project, an N-terminal labeling mass spectrometry method, terminal amine isotopic labeling of substrates (TAILS), was optimized for the in-depth analysis of PHG and validated using a test protease (trypsin) with known cleavage specificity. Gluten N-termini in test and control groups were labeled with heavy and light formaldehyde, respectively. Trypsin-generated neo N-termini were identified by exhibiting an MS1 Log2 H:L peak area ratio with a significant difference (p < .01) from zero and native N-termini with no significant difference from zero (p > .01). Using this strategy, all abundant, theoretical, test protease-generated peptides in exemplar alpha/beta gliadins and gamma gliadins were identified. SIGNIFICANCE: This study is the first study that modified and evaluated TAILS analysis for the analysis of partially hydrolyzed gluten proteins. The evaluation indicated that the TAILS analysis could be modified and expanded to the identification of multiple protease cleavage sites in gluten proteins and is worth further evaluation as a novel strategy for the analysis of natural hydrolysis of gluten in food processes. This strategy also may be further applied to characterize a broader range of partially hydrolyzed allergens in foods and provide reference for their safety assessment to both industry and regulatory authorities.

Detection of gluten in a pilot-scale barley-based beer produced with and without a prolyl endopeptidase enzyme.

Immunochemical and mass spectrometric methods were used to examine the gluten composition of a gluten-reduced beer produced by brewing with barley malt in the presence of prolyl endopeptidase (PEP) and a final filtration treatment with diatomaceous earth and perlite. The competitive ELISA is generally considered appropriate for the analysis of hydrolysed gluten, but it is not considered a scientifically valid method for the quantification of gluten in fermented or hydrolysed foods due to the lack of an appropriate reference standard. As no single analytical method can capture the spectrum of gluten-derived products in beer, a comprehensive approach was employed to analyse the intact and hydrolysed fractions of gluten with complementary methods. The combination of PEP addition and diatomaceous earth/perlite filtration was more effective at reducing the concentration of detectable gluten than each of the treatments alone. However, gluten proteins and/or polypeptides were observed in filtered, PEP-treated beers using sandwich ELISA methods, western blot, and bottom-up mass spectrometry. In addition, mass spectrometry results showed that the number of hydrolysed gluten peptides was almost unaffected by the filtration process. Gluten peptides that contained potentially immunopathogenic sequences were identified in the filtered PEP-containing beers by MS. Variability in gluten composition was observed between three replicate pilot-scale productions, suggesting that the gluten profile in beer could differ from batch to batch. As there is uncertainty in the detection and quantification of gluten in hydrolysed and fermented foods, characterisation of hydrolysed gluten by complementary analytical methodologies is recommended.

Detection of Gluten during the Fermentation Process To Produce Soy Sauce.

Advances have been made to provide people with celiac disease (CD) access to a diverse diet through an increase in the availability of gluten-free food products and regulations designed to increase label reliability. Despite advances in our knowledge regarding CD and analytical methods to detect gluten, little is known about the effects of fermentation on gluten detection. The enzyme-linked immunosorbent assay (ELISA) and lateral flow devices routinely used by analytical laboratories and regulatory agencies to test for the presence of gluten in food were examined for their ability to detect gluten during the fermentation processes leading to the production of soy sauce, as well as in finished products. Similar results were observed irrespective of whether the soy sauce was produced using pilot-plant facilities or according to a homemade protocol. In both cases, gluten was not detected after moromi (brine-based) fermentation, which is the second stage of fermentation. The inability to detect gluten after moromi fermentation was irrespective of whether the assay used a sandwich configuration that required two epitopes or a competitive configuration that required only one epitope. Consistent with these results was the observation that ELISA, lateral flow devices, and Western immunoblot analyses were unable to detect gluten in commercial soy sauce, teriyaki sauce, and Worcestershire sauce. Although reports are lacking on problems associated with the consumption of fermented soy-containing sauces by consumers with CD, additional research is needed to determine whether all immunopathogenic elements in gluten are hydrolyzed during soy sauce production.