ObjectiveTo clarify whether epigallocatechin gallate （EGCG） is involved in the clearance of amyloid β-protein （Aβ） and autophagy induction by microglia， so as to explore the potential mechanisms of EGCG in the prevention and treatment of Alzheimer's disease （AD）. MethodsSix-month-old APP/PS1 mice were randomly divided into model and EGCG groups， with some additional wild type （WT） mice as the control group， each group consisting of 15 mice. The EGCG group received continuous gavage administration［5 mg/（kg·d）］ for 8 weeks， followed by the open field test and Y-maze to assess the learning and memory abilities of the mice. Thioflavin-S staining was used to evaluate the content and distribution of amyloid β-protein （Aβ）in the brain parenchyma of the mice， and immunofluorescence was employed to detect the expression levels of Aβ_1-42， glial fibrillary acidic protein （GFAP）， and ionized calcium-binding adapter molecule 1 （Iba1） in the hippocampal tissue of the mice. Additionally， N9 mouse microglial cells were induced with 20 µmol/L Aβ_1-42， and the cell viability was measured after treatment with different concentrations of EGCG （5 µmol/L， 10 µmol/L， 20 µmol/L）. Western blotting was used to detect the levels of Aβ_1-42， low density lipoprotein receptor-related protein 1（LRP1）， receptor for advanced glycation endproducts （RAGE）， amyloid precursor protein （APP）， insulin degrading enzyme （IDE）， neprilysin （NEP）， microtubule associated protein 1 hydrogen chain 3（LC3）-Ⅱ/LC3-Ⅰ， phosphatidylinositol 3-hydroxy kinase（PI3K）， p-PI3K， protein kinase B （AKT）， p-AKT， mammalian target of rapamycin （mTOR）， p-mTOR， and histone deacetylase 6（HDAC6）. Finally， through the co-culture of microglial cells and neuronal SH-SY5Y cells， cell viability and Caspase-3 levels were measured to verify the protective effect of EGCG-mediated Aβ clearance on neurons. ResultsEGCG increased the activity time and frequency of APP/PS1 mice in the central area of the open field （P<0.05）， and enhanced the percentage of alternation in the Y-maze test （P<0.01）； EGCG reduced Aβ deposition in the hippocampal tissue of APP/PS1 mice and increased the number of microglia； in vitro experiments showed that EGCG improved the survival rate of Aβ-induced N9 cells （P<0.01）， upregulated RAGE activity （P<0.05）， and promoted the internalization and phagocytosis of Aβ （P<0.01）. ECGC activated microglial autophagy by downregulating the level of HDAC6 （P<0.05）， inhibiting the phosphorylation of PI3K， AKT， mTOR （P<0.001）， and increasing the LC3-Ⅱ/LC3-I ratio （P<0.001）； EGCG improved the survival rate of SH-SY5Y cells （P<0.05） and reduced the activity of Caspase-3 （P<0.01） by clearing Aβ_1-42 through microglia， and had a protective effect on neurons. ConclusionEGCG activates microglial autophagy to clear Aβ by targeting and inhibiting the HDAC6-PI3K/AKT/mTOR axis.

Non-small cell lung cancer is one of the cancers with the highest incidence and mortality rate in the world, and precise prognostic models can guide clinical treatment plans. With the continuous upgrading of computer technology, deep learning as a breakthrough technology of artificial intelligence has shown good performance and great potential in the application of non-small cell lung cancer prognosis model. The research on the application of deep learning in survival and recurrence prediction, efficacy prediction, distant metastasis prediction, and complication prediction of non-small cell lung cancer has made some progress, and it shows a trend of multi-omics and multi-modal joint, but there are still shortcomings, which should be further explored in the future to strengthen model verification and solve practical problems in clinical practice.

Objective To compare the discrepancies among results of three commonly used laboratory direct test methods for maximal oxygen uptake(VO2max),explore their linear regression relationships,mutual predictability and comparability.Methods Using a quasi-experimental design of cluster sampling and within-group interaction design,20 male cross-country skiers were tested for VO2max using the Bruce protocol(Method 1),90-second incremental load exercise on power bicycle(Method 2),and 1-minute incremental load exercise on treadmill(Method 3),with an interval of one week.The indepen-dent and dependent variable were the three VO2max test methods and the VO2max,respectively.Results Significant differences were found in the average VO2max of the three test results,with the value mea-sured by Method 1 ranking the first,followed by that assessed by Method 3 and Method 2(P<0.05).Moreover,the frequency of individual differences in the results of the three methods showed that the VO2max of Method 1 was about 6 and 3 ml/min·kg higher than that measured by Method 2 and 3.However,at the same treadmill speed,the average blood lactate evaluated using Method 3 was higher than Method 1,and the speed reached aerobic and anaerobic thresholds about one speed unit(1 km/h)lower than Method 1.Meanwhile,linear regression analyses of the test results between Method 1 and 2,as well as Method 1 and 3 showed that both the regression models and coefficients were statis-tically significant(P<0.001),with the R-squared values of 9.25 and 9.05,respectively.Conclusion The Bruce protocol performs best in assessing the maximum value of the athlete's VO2max phase,whose results have linear regression relationships with the other two methods,and can be used for pre-dicting their results.Moreover,athletes of different events and levels can choose different VO2max test methods accordingly.Lastly,the speed and heart rate ranges corresponding to the aerobic and anaero-bic thresholds can serve as an effective and convenient method to control the training intensity.

Background: Galangin, commonly employed in traditional Chinese medicine for its diverse medicinal properties, exhibits potential in treating inflammatory pain. Nevertheless, its mechanism of action remains unclear. Methods: Mice were randomly divided into 4 groups for 7 days: a normal control group, a galangin-treated (25 and 50 mg/kg), and a positive control celecoxib (20 mg/kg). Analgesic and anti-inflammatory effects were evaluated using a hot plate test, acetic acid-induced writhing test, acetic acid-induced vascular permeability test, formalininduced paw licking test, and carrageenan-induced paw swelling test. The interplay between galangin, transient receptor potential vanilloid 1 (TRPV1), NF-κB, COX-2, and TNF-α proteins was evaluated via molecular docking. COX- 2, PGE2, IL-1β, IL-6, and TNF-α levels in serum were measured using ELISA after capsaicin administration (200 nmol/L). TRPV1 expression in the dorsal root ganglion was analyzed by Western blot. The quantities of substance P (SP) and calcitonin gene-related peptide (CGRP) were assessed using qPCR. Results: Galangin reduced hot plate-induced licking latency, acetic acid-induced contortions, carrageenantriggered foot inflammation, and capillary permeability in mice. It exhibited favorable affinity towards TRPV1, NF- κB, COX-2, and TNF-α, resulting in decreased levels of COX-2, PGE2, IL-1β, IL-6, and TNF-α in serum following capsaicin stimulation. Galangin effectively suppressed the upregulation of TRPV1 protein and associated receptor neuropeptides CGRP and SP mRNA, while concurrently inhibiting the expression of NF-κB, TNF-α, COX-2, and PGE2 mRNA. Conclusions Galangin exerts its anti-inflammatory pain effects by inhibiting TRPV1 activation and regulating COX-2, NF-κB/TNF-α expression, providing evidence for the use of galangin in the management of inflammatory pain.

Auditory neuropathy spectrum disorder (ANSD) represents a variety of sensorineural deafness conditions characterized by abnormal inner hair cells and/or auditory nerve function, but with the preservation of outer hair cell function. ANSD represents up to 15% of individuals with hearing impairments. Through mutation screening, bioinformatic analysis and expression studies, we have previously identified several apoptosis-inducing factor (AIF) mitochondria-associated 1 (AIFM1) variants in ANSD families and in some other sporadic cases. Here, to elucidate the pathogenic mechanisms underlying each AIFM1 variant, we generated AIF-null cells using the clustered regularly interspersed short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system and constructed AIF-wild type (WT) and AIF-mutant (mut) (p.‍T260A, p.‍R422W, and p.‍R451Q) stable transfection cell lines. We then analyzed AIF structure, coenzyme-binding affinity, apoptosis, and other aspects. Results revealed that these variants resulted in impaired dimerization, compromising AIF function. The reduction reaction of AIF variants had proceeded slower than that of AIF-WT. The average levels of AIF dimerization in AIF variant cells were only 34.5%‍‒‍49.7% of that of AIF-WT cells, resulting in caspase-independent apoptosis. The average percentage of apoptotic cells in the variants was 12.3%‍‒‍17.9%, which was significantly higher than that (6.9%‍‒‍7.4%) in controls. However, nicotinamide adenine dinucleotide (NADH) treatment promoted the reduction of apoptosis by rescuing AIF dimerization in AIF variant cells. Our findings show that the impairment of AIF dimerization by AIFM1 variants causes apoptosis contributing to ANSD, and introduce NADH as a potential drug for ANSD treatment. Our results help elucidate the mechanisms of ANSD and may lead to the provision of novel therapies.
Humans ; Apoptosis Inducing Factor/metabolism* ; NAD/metabolism* ; Dimerization ; Apoptosis

Objective:To explore the early clinical efficacy of ultrasound visualized platelet-rich plasma (PRP) in the treatment of lower back myofascial pain syndrome (MPS) after sports injury.Methods:A prospective cohort study was conducted to analyze the clinical data of 32 patients with lower back MPS after sports injury, who were admitted to West China Hospital of Sichuan University from January 2023 to March 2023. Ultrasound-guided PRP injection into the erector spinalis or quadratus psoas muscles was used for treatment. Before treatment, at 24 hours, 2 weeks, and 4 weeks after treatment, pain and function were evaluated using visual analogue scale (VAS), McGill pain questionnaire (McGill), Roland Morris dysfunction questionnaire (RMDQ), and Oswestry dysfunction index (ODI). Before treatment and 4 weeks after treatment, the quality of life was evaluated using the short-form 36 item health survey questionnaire (SF-36). The adverse reactions were observed during treatment and follow-up.Results:A total of 32 patients with lower back MPS after sports injury were enrolled, including 10 males and 22 females; aged 12-68 years [(47.3±16.3)years]. All the patients were followed up for 4 weeks. Before and at 24 hours, 2 weeks, and 4 weeks after treatment, the VAS was 5.0(4.0, 6.0)points, 3.5(3.0, 4.8)points, 2.0(2.0, 3.0)points, and 2.0(1.3, 3.0)points, respectively; the McGill score was 9.0(7.0, 11.0)points, 7.0(5.0, 9.0)points, 4.0(3.0, 5.0)points, and 3.0(3.0, 5.0)points, respectively; the RMDQ score was 8.0(5.3, 10.8)points, 5.5(3.0, 8.0)points, 4.0(3.0, 5.8)points, and 3.0(2.0, 4.8)points, respectively; the ODI was 22.0(14.5, 30.0), 20.0(14.5, 25.5), 9.0(6.0, 16.0), and 8.0(4.5, 14.0), respectively. Compared with the values before treatment, the VAS, McGill score, and RMDQ score were significantly decreased at 24 hours, 2 weeks, and 4 weeks after treatment (all P<0.05); the ODI had no significant difference at 24 hours after treatment ( P>0.05), but it was significantly decreased at 2 and 4 weeks after treatment (all P<0.05). Compared with the values at 24 hours after treatment, the VAS, McGill score, RMDQ score and ODI further decreased at 2 weeks after treatment (all P<0.05). Compared with the values at 2 weeks after treatment, there was no significant difference in the VAS, McGill score, RMDQ score, or ODI at 4 weeks after treatment (all P>0.05). In the SF-36, the scores of physiological function [77.5(60.0, 93.8)points], physiological role [50.0(0.0, 100.0)points], body pain [64.0(44.5, 74.0)points], vitality [75.0(65.0, 78.8)points], social function [87.5(75.0, 100.0)points], emotional role [66.7(33.3, 100.0)points] and mental health [72.0(68.0, 83.0)points] before treatment were increased to 90.0(80.0, 98.8)points, 100.0(56.3, 100.0)points, 84.0(74.0, 84.0)points, 75.0(70.0, 80.0)points, 100.0(87.5, 112.5)points, 100.0(66.7, 100.0)points, and 76.0(68.0, 84.0)points after 4 weeks of treatment, respectively ( P<0.05 or 0.01). However, there was no significant difference in the general health status or health changes before and after treatment (all P>0.05). During treatment and follow-up, no adverse reactions such as redness, swelling, pain, or subcutaneous bleeding were observed. Conclusion:Ultrasound-guided PRP treatment can improve the early pain, lumbar mobility and quality of life of patients with lower back MPS after sports injury, with no presence of adverse reactions.

Auditory neuropathy spectrum disorder (ANSD) represents a variety of sensorineural deafness conditions characterized by abnormal inner hair cells and/or auditory nerve function, but with the preservation of outer hair cell function. ANSD represents up to 15％ of individuals with hearing impairments. Through mutation screening, bioinformatic analysis and expression studies, we have previously identified several apoptosis-inducing factor (AIF) mitochondria-associated 1 (AIFM1) variants in ANSD families and in some other sporadic cases. Here, to elucidate the pathogenic mechanisms underlying each AIFM1 variant, we generated AIF-null cells using the clustered regularly interspersed short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system and constructed AIF-wild type (WT) and AIF-mutant (mut) (p.T260A, p.R422W, and p.R451Q) stable transfection cell lines. We then analyzed AIF structure, coenzyme-binding affinity, apoptosis, and other aspects. Results revealed that these variants resulted in impaired dimerization, compromising AIF function. The reduction reaction of AIF variants had proceeded slower than that of AIF-WT. The average levels of AIF dimerization in AIF variant cells were only 34.5％?49.7％ of that of AIF-WT cells, resulting in caspase-independent apoptosis. The average percentage of apoptotic cells in the variants was 12.3％?17.9％, which was significantly higher than that (6.9％?7.4％) in controls. However, nicotinamide adenine dinucleotide (NADH) treatment promoted the reduction of apoptosis by rescuing AIF dimerization in AIF variant cells. Our findings show that the impairment of AIF dimerization by AIFM1 variants causes apoptosis contributing to ANSD, and introduce NADH as a potential drug for ANSD treatment. Our results help elucidate the mechanisms of ANSD and may lead to the provision of novel therapies.

Inflammatory myofibroblastic tumor (IMT)is a potentially or low-grade malignant mesenchymal neoplasm, which is rare in clinic. Renomedullary interstitial cell tumor(RICT) is a clinically rare benign renal tumor. The combination of these two diseases in one patient has not been reported. A 25-year-old female patient was admitted to the hospital due to left back pain for 12 days and hematuria for 1 week. MRI of kidneys showed a mass in the left renal pelvis, which was considered as renal pelvic carcinoma. Urine cytopathological examination was negative. Robot-assisted laparoscopic radical left nephroureterectomy was performed. There was no tumor recurrence or metastasis during the follow-up for more than 6 months after operation.

Objective: To explore the effect of cryoablation on apoptosis of lung adenocarcinoma cells. Methods: According to the district in the tumor following cryoablation, human lung adenocarcinoma H1299 cells were divided into four groups to construct the freezing model of lung cancer in cell level: group A, untreated cells, as the control group. In group B, the necrotic solution was added to the untreated cell culture medium(the cell suspension was frozen in a liquid nitrogen tank for 5 min, and centrifuged to obtain the supernatant after rewarming at 37 ℃).Group C, sublethal cells(cells were frozen at-80 ℃ for 7 min to mimic sublethal state). In group D, necrotic solution was added into sublethal cell culture medium. Cell apoptosis was observed by flow cytometry analysis 24 h later. The expression of apoptosis related molecules Bax and Bcl-2 was detected by qRT-PCR and Western blot. Lewis lung adenocarcinoma cell line was used to establish subcutaneous transplanted tumor model of C57 BL/6 mice. The successful model mice were randomly divided into two groups: sham operation group and argon-helium cryoablation group, with 4 mice in each group. After argon-helium cryoablation for 14 days, the tumor tissues were taken. The expression of apoptosis-related molecules Bax and Bcl-2 was detected by qRT-PCR and Western blot. Results: In the cell freezing model, compared with group A in the control group B,C and D groups could promote cell apoptosis(P<0.05), and group D had the highest apoptosis rate, which was significantly higher than other groups(P<0.05). Bothin vitroandin vivoexperiments showed that cryoablation could increase the expression of Bax mRNA and protein(P<0.05). Bcl-2 mRNA and protein expression decreasedin vivo(P<0.05). There was no significant change in the in cell freezing model. Conclusion Cryoablation can achieve effective ablation through cell apoptosis mechanism, which may be related to the reduction of Bcl-2/Bax.

Objective:To investigate the relationship between serum 25 hydroxyvitamin-D3, soluble advanced glycation end product receptor (sRAGE), nucleotide binding oligomerization domain like receptor 3 (NLRP3) mRNA and cognitive impairment in hypertensive intracerebral hemorrhage (HICH).Methods:143 patients with HICH treated in the Affiliated Hospital of Guangdong Medical University from July 2016 to July 2019 were selected as the research objects. Among the 143 patients with HICH, there were 68 patients with cognitive impairment (cognitive impairment group) and 75 patients without cognitive impairment (control group). The age, gender, amount of intracerebral hemorrhage, bleeding site, blood pressure, blood glucose and blood lipid of the two groups were counted, and the mRNA levels of 25 hydroxyvitamin-D3, sRAGE and NLRP3 were detected. Multivariate logistic regression analysis was used to analyze the risk factors of cognitive impairment in patients with HICH.Results:There were no significant differences in age, gender, smoking, education, bleeding site, diabetes rate, triglyceride (TG), total cholesterol (TC), low density lipoprotein cholesterol (LDL-C), and high density lipoprotein cholesterol (HDL-C) between cognitive dysfunction group and control group ( P>0.05); There were significant differences in bleeding volume and neurological function defect score (NIHSS) score between cognitive impairment group and control group ( P<0.05); The level of 25 hydroxyvitamin-D3 in cognitive impairment group was lower than that in control group ( P<0.05), and the expression level of NLRP3 mRNA was higher than that in control group ( P<0.05). There was no significant difference in sRAGE between the two groups ( P>0.05); Logistic regression analysis showed that the decrease of 25-hydroxyvitamin-D3 level, the increase of bleeding volume and NIHSS score were independent risk factors for cognitive impairment in HICH patients ( P<0.05). Conclusions:Decreased serum 25 hydroxyvitamin-D3 levels may increase the risk of cognitive impairment in patients with HICH.