Probiotic fermentation environment control under intelligent data monitoring.

Probiotic fermentation studies are vital in many areas, particularly when it comes to feeding applications. This work examines probiotic fermentation in oil tea crops. The assessment of tea saponin-degrading bacteria and optimization of fermentation conditions using fermented oil tea cake under natural conditions, screening out six strains with strong ability to degrade tea saponin; selection of the best tea saponin degradation strain L.2 and recognition of its morphological features and ITS sequence to obtain L.2 strain is Aspergillus Niger. Oil tea is rich in tea saponin. Aspergillus Niger degraded tea saponins in oil teacakes at a rate of 93.96 % under the ideal conditions of 31.3 oC, 103.5 h, and 4.57 mL of initial acid addition. This has been accomplished via solid-state fermentation of L.2 using single factor studies and surface response optimization experiments. Moreover, Aspergillus Niger degraded tea saponins in oil tea cakes at a rate of 93.96% at the ideal circumstances of 31.3 C, 103.5 h, and 4.57 mL of initial acid addition.

Platforms of graphene/MXene heterostructure for electrochemical monitoring of rutin in drug and Tartary buckwheat tea.

The use of two-dimensional heterostructure composite as electrode modification material has become a new strategy to improve the electrocatalytic activity and electroactive sites of electrochemical sensor. Herein, a soluble heterostructure, namely rGO-PSS@MXene, was designed and synthesized by integrating poly (sodium p-styrenesulfonate)-functionalized reduced graphene oxide into MXene nanosheets via ultrasonic method. The interactive heterostructure can effectively alleviate the self-stacking of MXene and rGO, endowing them with superior electron transfer capacity and large specific surface area, thereby producing prominent synergistic electrocatalytic effect towards rutin. In addition, the excellent enrichment effect of rGO-PSS@MXene for rutin also plays an important role through the electrostatic and π-π stacking interactions. The electrochemical characteristics of rutin on the sensor were examined in detail and a sensitive sensing method was proposed. Under optimized conditions, the method showed satisfactory linear relationship for rutin in the concentration range of 0.005-10.0 µM, with limit of detection of 1.8 nM (S/N = 3). The quantitative validation results in herbal medicine and commercial Tartary buckwheat tea were highly consistent with the labeled quantity and the results of HPLC determination, respectively, suggesting the sensor possessed excellent selectivity and accuracy. This proposed strategy for rutin determination is expected to expand the application of MXene heterostructure in electrochemical sensors, and is envisioned as a promising candidate for quality monitoring of drugs and foods.

Ginsenoside Rb2 inhibits p300-mediated SF3A2 acetylation at lysine 10 to promote Fscn1 alternative splicing against myocardial ischemic/reperfusion injury.

INTRODUCTION: Ginsenosides (GS) derived from Panax ginseng can regulate protein acetylation to promote mitochondrial function for protecting cardiomyocytes. However, the potential mechanisms of GS for regulating acetylation modification are not yet clear. OBJECTIVES: This study aimed to explore the potential mechanisms of GS in regulating protein acetylation and identify ginsenoside monomer for fighting myocardial ischemia-related diseases. METHODS: The 4D-lable free acetylomic analysis was employed to gain the acetylated proteins regulated by GS pretreatment. The co-immunoprecipitation assay, immunofluorescent staining, and mitochondrial respiration measurement were performed to detect the effect of GS or ginsenoside monomer on acetylated protein level and mitochondrial function. RNA sequencing, site-specific mutation, and shRNA interference were used to explore the downstream targets of acetylation modificationby GS. Cellular thermal shift assay and surface plasmon resonance were used for identifying the binding of ginsenoside with target protein. RESULTS: In the cardiomyocytes of normal, oxygen glucose deprivation and/or reperfusion conditions, the acetylomic analysis identified that the acetylated levels of spliceosome proteins were inhibited by GS pretreatment and SF3A2 acetylation at lysine 10 (K10) was significantly decreased as a potential target of GS. Ginsenoside Rb2 was identified as one of the active ginsenoside monomers for reducing the acetylation of SF3A2 (K10), which enhanced mitochondrial respiration against myocardial ischemic injury in in vivo and in vitro experiments. RNA-seq analysis showed that ginsenoside Rb2 promoted alternative splicing of mitochondrial function-related genes and the level of fascin actin-bundling protein 1 (Fscn1) was obviously upregulated, which was dependent on SF3A2 acetylation. Critically, thermodynamic, kinetic and enzymatic experiments demonstrated that ginsenoside Rb2 directly interacted with p300 for inhibiting its activity. CONCLUSION: These findings provide a novel mechanism underlying cardiomyocyte protection of ginsenoside Rb2 by inhibiting p300-mediated SF3A2 acteylation for promoting Fscn1 expression, which might be a promising approach for the prevention and treatment of myocardial ischemic diseases.

Improved Sleep Affects Epigastric Pain in Functional Dyspepsia by Reducing the Levels of Inflammatory Mediators.

INTRODUCTION: The pathogenesis of epigastric pain in functional dyspepsia (FD) is complex. The study aims to explore the effect of sleep improvement on this symptom. METHODS: In total, 120 patients with FD-associated epigastric pain and insomnia were randomly divided into experimental and control groups using the envelope method. After applying the exclusion criteria, 107 patients were enrolled in the experimental (56 patients) and control (51 patients) groups. Insomnia was graded according to the Pittsburgh Sleep Quality Index (PSQI). In the experimental group, eszopiclone 3 mg, eszopiclone 3 mg + estazolam 1 mg, and eszopiclone 3 mg + estazolam 2 mg were given to patients with mild, moderate, and severe insomnia, respectively. In the control group, patients were given 1, 2, or 3 tablets of vitamin B complex. Patient sleep quality was monitored with Sleepthing. Epigastric pain was evaluated with a Numeric Rating Scale. The serum levels of IL-1ß, IL-6, IL-8, and tumor necrosis factor-α (TNF-α) were measured by enzyme-linked immunosorbent assay. Pain scores, sleep parameters, and serum levels of inflammatory mediators were compared before and after treatment. RESULTS: After treatment, the pain scores, sleep parameters, and TNF-α and IL-6 levels in the experimental group were significantly lower than those in the control group (p < 0.05). PSQI insomnia scores were significantly associated with pain scores, IL-6, and TNF-α (p < 0.05) but not in IL-8 and IL-1ß levels (p > 0.05) among the three groups. CONCLUSIONS: Improving sleep with eszopiclone and/or estazolam alleviates FD-associated epigastric pain, possibly by inhibiting related downstream transmission pathways and reducing the release of inflammatory mediators.

Silicon Nanoparticle-Based Ratiometric Fluorescence Probes for Highly Sensitive and Visual Detection of VB2.

In this work, blue fluorescent silicon nanoparticles (SiNPs) were prepared by a simple one-step hydrothermal method using (3-aminopropyl) triethoxy silane (APTES) and eriochrome black T as raw materials. The SiNPs showed favorable water solubility, thermal stability, pH stability, salt tolerance, and photobleaching resistance. At an excitation wavelength of 376 nm, the SiNPs emitted bright blue fluorescence at 460 nm. In the presence of vitamin B2 (VB2), the fluorescence intensity (FL intensity) of the SiNPs at 460 nm decreased obviously, and a new peak appeared at 521 nm. Based on this, a novel ratiometric fluorescence method was established for VB2 detection. There was a good linear relationship between the fluorescence intensity ratio (F 521/F 460) and VB2 concentration from 0.5 to 60 µM with a detection limit of 135 nM. This method was successfully applied to detect VB2 content in the samples of vitamin B2 drugs and beverages. Additionally, a simple paper sensor based on the SiNPs was designed to visualize detection of VB2. With the support of color recognition software on a smartphone, the visual quantitative analysis of VB2 was realized, ranging from 40 to 800 µM.

Therapeutic application of natural products: NAD+ metabolism as potential target.

BACKGROUND: Nicotinamide adenine dinucleotide (NAD+) metabolism is involved in the entire physiopathological process and is critical to human health. Long-term imbalance in NAD+ homeostasis is associated with various diseases, including non-alcoholic fatty liver disease, diabetes mellitus, cardiovascular diseases, neurodegenerative disorders, aging, and cancer, making it a potential target for effective therapeutic strategies. Currently, several natural products that target NAD+ metabolism have been widely reported to have significant therapeutic effects, but systematic summaries are lacking. PURPOSE: To summarize the latest findings on the prevention and treatment of various diseases through the regulation of NAD+ metabolism by various natural products in vivo and in vitro models, and evaluate the toxicities of the natural products. METHODS: PubMed, Web of Science, and ScienceDirect were searched using the keywords "natural products sources," "toxicology," "NAD+ clinical trials," and "NAD+," and/or paired with "natural products" and "diseases" for studies published within the last decade until January 2023. RESULTS: We found that the natural products mainly include phenols (curcumin, cyclocurcumin, 4-hydroxybenzyl alcohol, salvianolic acid B, pterostilbene, EGCG), flavonoids (pinostrobin, apigenin, acacetin, tilianin, kaempferol, quercetin, isoliquiritigenin, luteolin, silybin, hydroxysafflor yellow A, scutellarin), glycosides (salidroside), quinones (emodin, embelin, ß-LAPachone, shikonin), terpenoids (notoginsenoside R1, ginsenoside F2, ginsenoside Rd, ginsenoside Rb1, ginsenoside Rg3, thymoquinone, genipin), pyrazines (tetramethylpyrazine), alkaloids (evodiamine, berberine), and phenylpropanoids (ferulic acid). These natural products have antioxidant, energy-producing, anti-inflammatory, anti-apoptotic and anti-aging effects, which mainly influence the NAMPT/NAD+/SIRT, AMPK/SIRT1/PGC-1α, Nrf2/HO-1, PKCs/PARPs/NF-κB, and AMPK/Nrf2/mTOR signaling pathways, thereby regulating NAD+ metabolism to prevent and treat various diseases. These natural products have been shown to be safe, tolerable and have fewer adverse effects in various in vivo and in vitro studies and clinical trials. CONCLUSION: We evaluated the toxic effects of natural products and summarized the available clinical trials on NAD+ metabolism, as well as the recent advances in the therapeutic application of natural products targeting NAD+ metabolism, with the aim to provide new insights into the treatment of multiple disorders.

[Governor vessel moxibustion combined with warming acupuncture for knee osteoarthritis with yang deficiency and cold congelation based on the supporting-yang theory].

OBJECTIVE: To compare the clinical efficacy between governor vessel moxibustion combined with warming acupuncture based on the supporting-yang theory and simple warming acupuncture for knee osteoarthritis (KOA) with yang deficiency and cold congelation. METHODS: A total of 64 patients with KOA of yang deficiency and cold congelation were randomized into a combination group (32 cases, 2 cases dropped off) and a warming acupuncture group (32 cases, 1 case dropped off). In the warming acupuncture group, warming acupuncture was applied at Zusanli (ST 36), Guanyuan (CV 4) and Dubi (ST 35), Neixiyan (EX-LE 4), etc. on the affected side, once a day. On the basis of the treatment in the warming acupuncture group, governor vessel moxibustion was applied in the combination group, once a week. The 14-day treatment was taken as one course, and totally 2 courses with 2-day interval were required in the two groups. The clinical symptom score, the visual analogue scale (VAS) score and the Western Ontario and McMaster Universities arthritis index (WOMAC) score were observed before treatment, after treatment and in the follow-up of 12 weeks after treatment; the volume of suprapatellar bursa effusion was detected before and after treatment; the clinical efficacy was evaluated after treatment and in the follow-up in the two groups. RESULTS: After treatment and in the follow-up, the scores of clinical symptom, VAS and WOMAC were decreased compared before treatment in both groups (P<0.05), and those in the combination group were lower than the warming acupuncture group (P<0.05). After treatment, the volume of suprapatellar bursa effusion was decreased compared before treatment in both groups (P<0.05). After treatment and in the follow-up, the total effective rates were 93.3% (28/30) and 86.7% (26/30) in the combination group, which were superior to 87.1% (27/31) and 74.2% (23/31) in the warming acupuncture group respectively (P<0.05). CONCLUSION: Governor vessel moxibustion combined with warming acupuncture can improve the clinical symptoms i.e. pain and dysfunction and reduce the volume of suprapatellar bursa effusion in KOA patients with yang deficiency and cold congelation, its short-term effect and long-term effect are both superior to simple warming acupuncture.

Effects of transcranial direct current stimulation for post-stroke depression: A systematic review and meta-analysis.

OBJECTIVE: The aim of this investigation was to evaluate the effect of transcranial direct current stimulation (tDCS) on post-stroke depression (PSD). METHODS: Databases were searched through February 2021. Randomized Controlled Trial(RCT) investigating the efficacy of tDCS in PSD using a standardized depression scale as outcome measure was included. The Cochrane Manual of Systematic Evaluation 5.1.0 bias risk assessment tool was used to assess the risk of bias. Effect sizes were calculated from extracted data and combined for an overall summary statistic. RESULTS: Eight studies involving 412 patients were included. These trials revealed a significant pooled effect size (standardized mean differences(SMD) = 1.61, 95% confidence interval(CI) 1.02-2.19, P < 0.00001). For the subgroup analyses, neither comparisons of high- vs low-intensity or left dorsolateral prefrontal cortex vs primary motor cortex revealed a significant result. tDCS had no obvious effect on the anxiety state in patients with PSD(SMD = 1.09, 95% CI, -0.22 to 2.40, P = 0.10), while it resulted in improved ability of daily life(ADL) in these patients(SMD = 0.82, 95% CI, 0.16-1.48, P = 0.01) CONCLUSIONS: tDCS has an effect on improvement in PSD, while it is still not clear which stimulus program is best. SIGNIFICANCE: Further trials with larger sample sizes are needed to determine the best and most effective tDCS regimen and to realize potential clinical applications.

Associations between food-specific IgG and health outcomes in an asymptomatic physical examination cohort.

BACKGROUND: Although the association of food-specific IgG with the development and progression of specific diseases was shown by many studies, it is also present in the population without clinical symptoms. However, the association between food-specific IgG and physical examination outcomes in healthy people has not been studied yet. METHODS: An asymptomatic physical examination cohort (APEC) was selected according to the inclusion and exclusion criteria, the physical examination data were compared between IgG positive and IgG negative groups, and their odds ratios (ORs) and 95% confidence intervals (CIs) were calculated using multivariable logistic regression. RESULTS: The data of 28,292 subjects were included in the analysis. The overall IgG positive rate was up to 52.30%, mostly with mild to moderate IgG positivity. The multivariable Logistic regression showed the prevalence of hypertriglyceridemia, abnormal fasting blood glucose and overweight was lower in the IgG (+) positive group (OR 0.87, 95% CI 0.83-0.92; OR 0.93, 95% CI 0.87-0.99; OR 0.92, 95% CI 0.87-0.96) but there was a higher prevalence of thyroid disease (OR 1.09, 95% CI 1.04-1.15). CONCLUSION: Food-specific IgG positivity was widespread in the APEC and was associated with lower prevalence of hypertriglyceridemia, abnormal fasting blood glucose and overweight. The underlying physiological mechanism merits further study.

Effect of Holding Time on the Extrusion Force and Microstructure Evolution during the Plastic Forming of Ti-6Al-4V Micro-Gears.

The application of titanium alloy micro-gears in microelectromechanical systems has been severely restricted, as the graphite mold is prone to abrasion or even to crack at high temperatures, mainly due to the forming load. We aimed to manufacture Ti-6Al-4V alloy micro-gears through hot extrusion under an electric field and to clarify the influence of holding time on the extrusion force. The results suggest that the formed gears had a complete filling and clear tooth profile. Moreover, the contact resistance and current density caused a gradient temperature distribution inside the billet, resulting in a carburized layer and inhomogeneous ß grains. The extrusion force increased with an increased holding time, which can be ascribed to the increase in the thickness of the carburized layer and the ß grain size. Among these two factors, ß grain size played a leading role in the extrusion force. Continuous dynamic recrystallization dominated the deformation in a single ß phase, and the misorientation of the transformed α laths from ß grains followed the Burgers orientation relationship. This study may pave the way for the extrusion forming of other titanium alloy micro-components.

[Panlong acupuncture combined with Xingnao Kaiqiao acupuncture for post-stroke fatigue: a randomized controlled trial].

OBJECTIVE: To observe the clinical effect of Panlong (dragon-like) acupuncture combined with Xingnao Kaiqiao (regaining consciousness and opening orifice) acupuncture on post-stroke fatigue. METHODS: Sixty patients with post-stroke fatigue were randomly divided into an observation group (30 cases, 1 case dropped off) and a control group (30 cases, 2 cases dropped off). The patients in the control group were treated with Xingnao Kaiqiao acupuncture at Neiguan (PC 6), Shuigou (GV 26), etc. for 30 min; based on the treatment of the control group, the patients in the observation group were treated with Panlong acupuncture at Jiaji (EX-B 2) of T1 to L5 for 15 min. The two groups were treated once a day, 6 times a week, for a total of 4 weeks. The scores of fatigue assessment instrument (FAI), energy of stroke-specific quality of life (SS-QOL), and modified Barthel index (MBI) in the two groups were compared before and after treatment, and the clinical efficacy was evaluated. RESULTS: Compared before treatment, the total score and each item score of FAI in the observation group were reduced after treatment (P<0.05), while the total score and FAI-1 and FAI-4 scores in the control group were reduced after treatment (P<0.05). The total score of FAI and FAI-1, FAI-2 and FAI-4 scores in the observation group were lower than those in the control group (P<0.05). After treatment, the SS-QOL energy scores and MBI scores of the two groups were increased compared before treatment (P<0.05), and the SS-QOL energy score in the observation group was higher than that in the control group (P<0.01). The total effective rate was 72.4% (21/29) in the observation group, which was better than 46.4% (13/28) in the control group (P<0.05). CONCLUSION: The Panlong acupuncture combined with Xingnao Kaiqiao acupuncture could effectively alleviate the fatigue symptoms and improve quality of life in patients with post-stroke fatigue.

[Acupuncture adjuvant treatment for dysphagia in patients with Parkinson's disease: a randomized controlled trial].

OBJECTIVE: To observe the effect of acupuncture on swallowing function and nutritional status of patients with Parkinson's disease (PD) dysphagia. METHODS: A total of 56 patients with PD dysphagia were randomly divided into an observation group and a control group, 28 cases in each one. Both groups were given conventional treatment and swallowing function rehabilitation training.On the basis, the observation group was treated with acupuncture (filiform needling and tongue picking acupuncture). The acupoints of filiform needling were Lianquan (CV 23), Shanglianquan (Extra), Yifeng (TE 17), etc.; and the tongue picking acupuncture was applied at Jinjin (EX-HN 12), Yuye (EX-HN 13) and posterior pharyngeal wall. Rehabilitation training and acupuncture were performed once a day for 5 consecutive days a week, 2 weeks as a course of treatment, 3 courses in total. The swallowing function [oral transit time (OTT), swallowing response time (SRT), pharyngeal transit time (PTT), laryngeal closure duration (LCD)] and nutritional indexes [body mass index (BMI), serum albumin (ALB), serum prealbumin (PA), hemoglobin (Hb)] in the two groups were evaluated before and after treatment, and the clinical effects were compared. RESULTS: After treatment, the paste and liquid OTT, SRT, PTT, LCD in the two groups were shorter than those before treatment (P<0.05), and the observation group was shorter than the control group (P<0.05). After treatment, the nutritional indexes (BMI, ALB, PA, Hb) in the two groups were higher than those before treatment (P<0.05), and the observation group was higher than the control group (P<0.05). The total effective rate of the observation group was 92.9% (26/28), which was better than 67.9% (19/28) of the control group (P<0.05). CONCLUSION: Acupuncture-assisted rehabilitation training can improve the swallowing function and nutritional status in patients with Parkinson's disease swallowing disorders.

A FRET-based ratiometric fluorescent probe for highly selective detection of hydrogen polysulfides based on a coumarin-rhodol derivative.

In modern biology, hydrogen polysulfides (H2Sn, n > 1) are members of reactive sulfur species (RSS), with anti-oxidation, cell protection and redox signals in tissues and organs. Therefore, it is crucial to develop a method to monitor the changes of H2Sn level in organisms. We designed and synthesized a ratiometric fluorescent probe for highly selective detection of H2Sn based on the fluorescence resonance energy transfer (FRET) process. In this work, a coumarin derivative was chosen as an energy donor, a rhodol derivative was used as an energy acceptor and a 2-fluoro-5-nitrobenzoate group was applied as a recognition unit for H2Sn. In the absence of H2Sn, the rhodol receptor existed in the non-fluorescent spirolactone state and FRET process was disabled. In the presence of H2Sn, the closed spirolactone form was converted to a conjugated fluorescent xanthenes form to invoke the occurrence of FRET which resulted in a 77 nm red-shift of fluorescence emission from 460 nm to 537 nm. The ratio value of the fluorescence intensity between 537 nm and 460 nm (I537nm/I460nm) of the probe exhibited a good linear relationship toward H2Sn in the range of 3.0 × 10-6-1.0 × 10-4 mol·L-1, and the detection limit was estimated to be 8.0 × 10-7 mol·L-1. In addition, the ratiometric fluorescent probe showed high specificity for H2Sn over other biologically related species. Moreover, the probe displayed little cell toxicity and had been successfully used to the confocal imaging of H2Sn in HepG2 cells by dual emission channels.

Advances in the discovery of microRNA-based anticancer therapeutics: latest tools and developments.

Introduction: MicroRNAs (miRNAs) are small endogenous non-coding RNAs that repress the expression of their target genes by reducing mRNA stability and/or inhibiting translation. miRNAs are known to be aberrantly regulated in cancers. Modulators of miRNA (mimics and antagonists) have emerged as novel therapeutic tools for cancer treatment.Areas covered: This review summarizes the various strategies that have been applied to correct the dysregulated miRNA in cancer cells. The authors also discuss the recent advances in the technical development and preclinical/clinical evaluation of miRNA-based therapeutic agents.Expert opinion: Application of miRNA-based therapeutics for cancer treatment is appealing because they are able to modulate multiple dysregulated genes and/or signaling pathways in cancer cells. Major obstacles hindering their clinical development include drug delivery, off-target effects, efficacious dose determination, and safety. Tumor site-specific delivery of novel miRNA therapeutics may help to minimize off-target effects and toxicity. Combination of miRNA therapeutics with other anticancer treatment modalities could provide a synergistic effect, thus allowing the use of lower dose, minimizing off-target effects, and improving the overall safety profile in cancer patients. It is critical to identify individual miRNAs with cancer type-specific and context-specific regulation of oncogenes and tumor-suppressor genes in order to facilitate the precise use of miRNA anticancer therapeutics.

Internet Addiction and Depression in Chinese Adolescents: A Moderated Mediation Model.

Research has revealed that Internet addiction is a risk factor for adolescents' development of depressive symptoms, although the underlying mechanisms are largely unknown. The present study examines the mediating role of positive youth development and the moderating role of mindfulness to determine the association between Internet addiction and depression. A sample of 522 Chinese adolescents completed measures related to Internet addiction, positive youth development, mindfulness, depression, and their background information, for which the results reveal that positive youth development mediates the relation between Internet addiction and depression. Moreover, the associations between both Internet addiction and depression as well as positive youth development and depression are moderated by mindfulness. These two effects were stronger for adolescents with low mindfulness than for those with high mindfulness. The present study contributes to a more thorough understanding of how and when Internet addiction increases the risk of depression in adolescents, suggesting that Internet addiction may affect adolescent depression through positive youth development and that mindfulness can alleviate the negative effect of Internet addiction or a low level of psychological resources on depression. The implications for research and practice are finally discussed.

The SGLT2 inhibitor dapagliflozin attenuates the activity of ROS-NLRP3 inflammasome axis in steatohepatitis with diabetes mellitus.

BACKGROUND: Diabetes mellitus (DM) is considered as a risk factor for the progress of liver diseases. After tissue damage, there is the highest amplitude of ubiquitously sterile inflammatory response in the liver, resulting in a major clinical consequence concerning a high prevalence of steatohepatitis in DM patients. This study aimed to investigate the inhibitory efficacy of dapagliflozin (DAPA), a sodium glucose cotransporter-2 (SGLT2) inhibitor, on experimental steatohepatitis with DM. METHODS: DM-steatohepatitis model was established by dual intraperitoneal injection of streptozotocin (STZ) and feeding with the high-fat diet (HFD) in apolipoprotein E-deficient (ApoE-/-) mice (n=40). The mice were concurrently treated with DAPA (1 mg/kg/d) by gavage for 12 weeks. RESULTS: In ApoE-/- mice, dual HFD/STZ dramatically induced hepatic damage and inflammation as compared with HFD alone. DAPA treatment was effective to protect from hepatic damage and inflammation in dual HFD/STZ treated ApoE-/- mice. DAPA also significantly the probability decreased the blood glucose, hepatic lipid accumulation, liver steatosis, and fibrotic response in dual HFD/STZ treated ApoE-/- mice. Further mechanistic investigations indicated that the protection of DAPA on diabetic liver injury was associated with the suppressed production of hepatic reactive oxygen species (ROS) and malondialdehyde (MDA) and the inhibited activation of NOD-like receptor family pyrin domain containing 3 (NLRP3) inflammasome. CONCLUSIONS: These data demonstrate the efficacy of DAPA for protecting liver damage, inflammation and steatosis from experimental steatohepatitis with DM, and indicate a possible involvement of the inhibited activity of ROS-NLRP3 inflammasome.

[Effect of electroacupuncture at Wnt/ß-catenin signaling pathway on inhibiting cartilage degeneration in rats with knee osteoarthritis].

OBJECTIVE: To explore the action mechanism of electroacupuncture (EA) for knee osteoarthritis (KOA) based on Wnt/beta-catenin (Wnt/ß-catenin) signaling pathway. METHODS: Ten rats were randomly selected into a sham-operation group among 50 male 2-month-old SD rats, and the KOA model was established in the remaining 40 rats by modified Hulth method. Four weeks after the model establishment, the rats were randomly divided into a model group, an experimental A group, an experimental B group and an experimental C group, 10 rats in each group. The rats in the sham-operation group and model group did not receive any intervention. The rats in the experimental A group were treated with EA at "Neixiyan" (EX-LE 4) and "Dubi"(ST 35) for 15 min. The rats in the experimental B group were treated with EA at "Neixiyan" (EX-LE 4) and "Dubi"(ST 35) for 30 min. The rats in the experimental C group were treated with EA at non-acupoint for 15 min. EA intervention was given once a day, five times a week, and totally 12-week treatment was given. After 12 weeks, the knee cartilage tissues were stained and the morphological changes were observed under light microscopy; the severity of cartilage degeneration was evaluated by modified Mankin's score; the content of interleukin-1ß (IL-1ß) in synovium tissues was detected by ELISA method; the content of Wnt-4, ß-catenin and matrix metalloprotein-13 (MMP-13) in cartilage tissues was detected by Western blot method. RESULTS: Compared with the sham-operation group, in the model group the morphology and structure of cartilage were disordered, the number of cells was significantly reduced, the matrix was decontaminated and tidal line was incomplete; the Mankin's score was significantly increased (P<0.01), the content of IL-1ß in synovium tissues were significantly increased (P<0.01), and the expressions of Wnt-4, ß-catenin and MMP-13 at protein level were significantly increased (P<0.01). Compared with the model group, in the experimental A group and experimental B group the morphology and structure of cartilage were more orderly, the number of cells was increased, the matrix staining was deepened and the tidal line was more complete; Mankin's scores were decreased significantly (P<0.01); the contents of IL-1ß in synovium tissues were decreased significantly (P<0.01), and the expressions of Wnt-4, ß-catenin and MMP-13 at protein level were decreased significantly (P<0.01). Compared with the model group, no improvement was observed in the experimental C group. Compared with the experimental A group, in the experimental C group the morphology and structure of cartilage were disordered, the number of cells was significantly reduced, the matrix was decontaminated and the tidal line was incomplete; Mankin's score was significantly increased (P<0.01), the content of IL-1ß in synovium tissues was significantly increased (P<0.01), and the expressions of Wnt-4, ß-catenin and MMP-13 at protein level were significantly increased (P<0.01). The morphological structure of cartilage in the experimental B group was similar to that in the experimental A group, and there was no significant difference in Mankin's score, IL-1ß content in synovium tissues and the expressions of Wnt-4, ß- catenin and MMP-13 at protein level between the two groups (P>0.05). CONCLUSION: EA at "Neixiyan" (EX-LE 4) and "Dubi"(ST 35) may reduce the expression of MMP-13 and the production of inflammatory factor IL-1ß through Wnt/ß-catenin signaling pathway, thus inhibit the degradation of cartilage matrix and the apoptosis of chondrocyte, and improve the morphology and structure of cartilage.

The Reduced Expression of EOLA1 May Be Related to Refractory Diabetic Foot Ulcer.

BACKGROUND: Chronic diabetic foot ulcer (DFU) is one of the most intractable complications of diabetes mellitus (DM). Its pathogenesis is complex, and uncontrolled chronic inflammation is an important factor. Endothelial overexpressed lipopolysaccharide-associated factor 1 (EOLA1) discovered in our laboratory is an intracellular protein with the function of inflammatory regulation. This study was aimed at observing the expression of EOLA1 in DFU skin tissues and its relationship with inflammation and at exploring the possible role of EOLA1 in DFU and its mechanism. METHODS: The patients with DFU were divided into 2 groups based on the formation time of ulcer: the acute wound (AW) group with the course of disease ≤ 4 weeks and the chronic wound (CW) group with the course of disease > 4 weeks. The relevant clinical data of patients were collected, and the skin tissues around the ulcer were used for immunofluorescence detection and immunohistochemical staining to observe inflammation. The expression levels of EOLA1, metallothionein 2A (MT2A), nuclear factor-κB (NF-κB), and interleukin-6 (IL-6) were detected by western blot. RESULTS: A total of 79 patients were enrolled in the study. The results of immunofluorescence and immunohistochemistry showed that EOLA1 was expressed in the epithelial tissues of DFU. However, the expression of EOLA1 in the CW group was significantly lower than that in the AW group (P < 0.05), and the expression of NF-κB and IL-6 was obviously increased (P < 0.05). CONCLUSION: The refractory wounds in patients with DFU may be closely related to the uncontrolled activation of inflammatory pathways in cells caused by the reduced expression of negative regulators of inflammation (e.g., EOLA1), and such decreased expression may be also strongly linked to the persistent state of inflammation.

Electroacupuncture Delays Cartilage Degeneration by Modulating Nuclear Factor-κB Signaling Pathway.

OBJECTIVE: To illustrate the molecular mechanisms underlying the therapeutic effects of electroacupuncture (EA) on knee osteoarthritis (OA). METHODS: Twenty-seven six-month-old New Zealand white rabbits were allocated into three groups in accordance with a random number table: normal group (no surgery-induced OA; without treatment), model group (surgery-induced OA; without treatment) and EA group [surgery-induced OA; received treatment with EA at acupoints Dubi (ST 35) and Neixiyan (EX-LE 5), 30 min twice a day]. After eight consecutive weeks of treatment, the histopathological alterations in cartilage were observed using optical microscopy and transmission electron microscopy, cartilage degeneration was evaluated by modified Mankin's score principles, the synovial fluid concentration of interleukin-1ß (IL-1ß), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and matrix metalloproteinase-3 (MMP-3) were evaluated by enzyme-linked immunosorbent assay, and the protein expression levels of IL-1ß, IL-6, TNF-α, MMP-3, IκB kinase-ß (IKK-ß), nuclear factor of α light polypeptide gene enhancer in B-cells inhibitor α (IκB-α) and nuclear factor-κB (NF-κB) p65 were quantified by Western blot analysis. RESULTS: EA treatment significantly improved cartilage structure arrangement and reduced cellular degeneration. The IL-1ß, IL-6, TNF-α and MMP-3 of synovial fluid in the EA-treated group were significantly decreased compared with the model group (all P<0.01). Compared with the model group, the IL-1ß, IL-6, TNF-α, MMP-3, IKK-ß and NF-κB p65 protein expressions in cartilage of EA-treated group were significantly decreased (all P<0.01), whereas IκB-α expression was significantly up-regulated (P<0.01). CONCLUSION: EA treatment may delay cartilage degeneration by down-regulating inflammatory factors through NF-κB signaling pathway, which may, in part, explain its clinical efficacy in the treatment of knee OA.

[Efficacy of acupuncture combined with hydro-acupuncture for transverse processes syndrome of the third lumbar vertebra].

OBJECTIVE: To observe the clinical efficacy differences between acupuncture combined with hydro-acupuncture and acupuncture alone for transverse processes syndrome of the third lumbar vertebra. METHODS: One hundred patients were randomly divided into an observation group and a control group, 50 cases in each one. The patients in the control group were treated with regular acupuncture at Weizhong (BL 40), ashi points, Jiaji (L2～L4) and Shenshu (BL 23); the acupuncture was given once a day, six treatments per week for consecutive 2 weeks. Based on the acupuncture treatment in the control group, in the observation group the No.5 injection needle was applied to relieve the tenderness points on the transverse process and articular process of the third lumbar vertebra, followed by hydro-acupuncture with Danshen injection (1 mL per point, 7 days per injection for 2 weeks). The visual analogue scale (VAS), Japanese Orthopaedic Association (JOA) scale and Oswestry disability index (ODI) were observed before treatment, after treatment and during six-month follow-up visit in the two groups. In addition, the clinical efficacy was compared between the two groups. RESULTS: Forty-eight patients completed treatment and 2 patients dropped off in the observation group, while 47 patients completed treatment and 3 patients dropped off in the control group. The total effective rate was 91.7% (44/48) in the observation group, which was significantly superior to 76.6% (36/47) in the control group (P<0.05). Compared before treatment, the VAS and ODI in the two groups were reduced after treatment and during six-month follow-up visit, while the JOA was increased (all P<0.05); the VAS and ODI in the observation group was lower than that in the control group, while JOA in the observation group was higher than that in the control group (all P<0.05). CONCLUSION: Acupuncture combined with hydro-acupuncture are effective for transverse processes syndrome of the third lumbar vertebra, and could significantly improve lumbar function and relieve the pain symptoms, which is superior to regular acupuncture.