OBJECTIVE: To observe the clinical efficacy of Qihuang needle therapy for autism spectrum disorder (ASD) children with sleep disorder. METHODS: A total of 60 ASD children with sleep disorder were randomly divided into an observation group and a control group, 30 cases in each group. Both groups were treated with structured education intervention, 60 min each time, once a day, 6 times a week. Qihuang needle therapy was applied at Yintang (GV24⁺), Baihui (GV20) and bilateral Jueyinshu (BL14), Xinshu (BL15) in the observation group, multi-direction needling was delivered and without needle retaining. The treatment was given 2 times a week, each treatment was delivered at interval of 2 days at least. Behavioral intervention was adopted in the control group. Treatment for consecutive 12 weeks was required in both groups. Before and after treatment, the scores of children's sleep habits questionnaire (CSHQ), the autism behavior checklist (ABC), the childhood autism rating scale (CARS), and the childhood autism behavior scale (CABS) were observed in the two groups. RESULTS: After treatment, the scores of CSHQ, ABC, CARS and CABS were decreased compared with those before treatment (P<0.01), and the above scores in the observation group were lower than those in the control group (P<0.05). CONCLUSION Qihuang needle therapy can effectively treat ASD with sleep disorder, improve the core symptoms of ASD and the sleep quality.
Humans ; Autism Spectrum Disorder/physiopathology* ; Male ; Female ; Child ; Sleep Wake Disorders/physiopathology* ; Child, Preschool ; Acupuncture Therapy ; Acupuncture Points ; Treatment Outcome ; Sleep ; Needles

Although clinical evidence suggests that nonalcoholic fatty liver disease is an established major risk factor for heart failure, it remains unexplored whether sleep disorder-caused hepatic damage contributes to the development of cardiovascular disease (CVD). Here, our findings revealed that sleep fragmentation (SF) displayed notable hepatic detrimental phenotypes, including steatosis and oxidative damage, along with significant abnormalities in cardiac structure and function. All these pathological changes persisted even after sleep recovery for 2 consecutive weeks or more, displaying memory properties. Mechanistically, persistent higher expression of nicotinamide adenine dinucleotide phosphate oxidase 4 (NOX4) in the liver was the key initiator of SF-accelerated damage phenotypes. SF epigenetically controlled the acetylation of histone H3 lysine 27 (H3K27ac) enrichment at the Nox4 promoter and markedly increased Nox4 expression in liver even after sleep recovery. Moreover, fine coordination of the circadian clock and hepatic damage was strictly controlled by BMAL1-dependent Sirtuin 1 (Sirt1) transcription after circadian misalignment. Accordingly, genetic manipulation of liver-specific Nox4 or Sirt1, along with pharmacological intervention targeting NOX4 (GLX351322) or SIRT1 (Resveratrol), could effectively erase the epigenetic modification of Nox4 by reducing the H3K27ac level and ameliorate the progression of liver pathology, thereby counteracting SF-evoked sustained CVD. Collectively, our findings may pave the way for strategies to mitigate myocardial injury from persistent hepatic detrimental memory in diabetic patients.

A major obstacle in type 2 diabetes mellitus (T2DM) is sleep fragmentation (SF), which negatively affects testicular function. However, the underlying mechanisms remain to be elucidated. In this study, we demonstrate that SF induces testicular damage through a mechanism involving lipid metabolism, specifically mediated by melatonin (MEL) receptor 1a (MT1). T2DM mice with SF intervention displayed several deleterious phenotypes such as apoptosis, deregulated lipid metabolism, and impaired testicular function. Unexpectedly, sleep recovery (SR) for 2 consecutive weeks could not completely abrogate SF's detrimental effects on lipid deposition and testicular function. Interestingly, MEL and MT1 agonist 2-iodomelatonin (2IM) effectively improved lipid homeostasis, highlighting MEL/2IM as a promising therapeutic drug for SF-trigged testicular damage. Mechanistically, MEL and 2IM activated FGFR1 and sequentially restrained the crosstalk and physical interaction between TAB1 and TAK1, which ultimately suppressed the phosphorylation of TAK1 to block lipid deposition and cell apoptosis caused by SF. The ameliorating effect of MEL/2IM was overtly nullified in Fgfr1 knockout (Fgfr1-KO ^+/- ) diabetic mice. Meanwhile, testicular-specific overexpression of Tak1 abolished the protective effect of FGF1^mut on diabetic mouse testis. Our findings offer valuable insights into the molecular mechanisms underlying the testicular pathogenesis associated with SF and propose a novel therapeutic approach for addressing male infertility in T2DM.

Objective To investigate the role of bufalin(BU)in inhibiting M2-type macrophage-mediated colorec-tal cancer metastasis.Methods Human acute leukemia mononuclear cells(THP-1)were differentiated into M0 macrophages using phorbol ester induction(PMA)for 48 hours.The M0 macrophages were then treated with IL-4 and IL-13 medium.Surface markers and morphological changes were observed through ELISA,morphology,and RT-qPCR experiments.RT-PCR and ELISA experiments were conducted to detect the surface markers TGF-β and IL-10 of M2 macrophages.The secretion level of IL-6 in the supernatant of M2 macrophages and colorectal cancer cells HCT116 was compared using ELISA.Additionally,the effect of conditioned medium on colorectal cancer cell HCT116 was assessed through Transwell,Wound healing,RT-qPCR,and Western blot experiments.Subsequent-ly,bufalin was added to the conditioned medium and the changes in AKT/PI3K protein,migration,and epithelial-mesenchymal transition ability in HCT116 were observed using Western blot,Transwell,Wound healing and RT-qPCR experiments.Results THP-1 were successfully differentiated into M2 macrophages.The activation of AKT/PI3K protein in HCT116 cells was induced by the secretion of IL-6 from M2 macrophages,which in turn promoted the migration and epithelial-mesenchymal transition ability of the HCT116 cells.The migration and epithelial-mes-enchymal transition mediated by M2 macrophages in HCT116 cells were effectively inhibited by Bufalin.Conclu-sion The release of IL-6 from M2 macrophages activates the AKT/PI3K signaling pathway in colorectal cancer cells,thereby promoting their migration and epithelial-mesenchymal transition capacity.Moreover,bufalin exhibits inhibitory effects on this effect.

Objective:To explore genes related to costimulatory molecule related to the prognosis of bladder cancer,and to construct and evaluate prognosis model based on costimulatory molecule-based signature(CMS).Methods:Gene expression matrix and clinical information of bladder cancer patients were downloaded from TCGA database and GEO database(GSE31684),and costimulatory molecule-related genes were retrieved from the literature.The univariate and multivariate Cox analysis were used to screened prognostic-related genes and constructed prognostic model.Forecast accuracy of model was verified in TCGA training group,TCGA validation data group and GEO group by Kaplan-Meier survival analysis and receiver operating characteristic curve(ROC).Considering risk score and clinical characteristics,we constructed a nomogram and evaluated its performance by consistency analysis and ROC.CIBERSORT algorithm was used to analyze immune cell composition of tumor microenvironment infiltration,and gene set enrichment analysis(GSEA)was performed to explore the potential mechanism.Results:Four prognostic-related CMSs were found:TNFRSF14,CD276,ICOS and TMIGD2,of which three were included in the risk score construction.Multivariate Cox regression results showed that the risk score based on CMS was an independent prognostic factor for bladder cancer patients.Consistency analysis and ROC results showed that the nomogram had ideal prognosis prediction accuracy.Immune infiltration analysis showed that the high risk group was likely to be in immunosuppressive state.GSEA results suggested that genes in high risk group were enriched in extracel-lular matrix(ECM)receptors interaction,cell cycle and other pathways.Conclusion:TNFRSF14,CD276 and ICOS may be potential prognostic biomarkers for bladder cancer patients.CMS-based risk score and nomogram could contribute to early prognosis and choice of personalized treatment.

Objective To analyze thein vivo three-dimensional dose verification using electronic portal imaging device(EIVD)for intensity-modulated radiotherapy(IMRT)of cervical cancer for investigating the differences between the measured and planned doses,and explore the optimal threshold for gamma passing rate in EIVD quality control based on dosimetric sensitivity.Methods A retrospective analysis was conducted on a cohort of 45 patients with cervical cancer who underwent IMRT at Women's Hospital,School of Medicine,Zhejiang University.During the treatment,all patients underwent EIVD to obtain the measured doses.The passing rate was analyzed using global gamma criteria of 2 mm/2%,2 mm/3%,and 3 mm/3%.Additionally,dose-volume histogram parameters were utilized to evaluate any differences between the measured and planned doses.Pearson correlation analysis was employed to investigate the relationship between the gamma passing rate and dosimetric differences.Furthermore,receiver operating characteristic(ROC)curve was generated to determine the optimal threshold for the gamma passing rate.Results The average gamma passing rates for the criteria of 2 mm/2%,2 mm/3%,and 3 mm/3%were 83.07%±5.25%,91.69%±3.52%,and 95.02%±2.46%,respectively.The Dmean deviation between EIVD measurement and planned dose in the planning target area was 2.43%(P=0.016),while the Dmean deviations in the bladder,rectum,and small intestine were 0.35%,0.46%,and 0.30%,respectively(P>0.05).Pearson analysis revealed a strong correlation between the 3 gamma indexes and dosimetric differences in the PTV(r>0.7),but a weak correlation with organs-at-risk(r<0.7).ROC analysis indicated that the optimal gamma passing rate thresholds for the criteria of 2 mm/2%,2 mm/3%,and 3 mm/3%were 79.06%,90.04%,and 94.19%,respectively.Conclusion The implementation of EIVD can ensure the accuracy of dose delivery within the PTV during IMRT for cervical cancer.Moreover,establishing a gamma passing rate threshold provides a valuable clinical basis for subsequent adaptive IMRT for cervical cancer.

Severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)is the main pathogen that causes COVID-19,which is fast-mutating and highly transmissible.The infection has led to a global epidemic.As the main preventive and control measure,vaccination plays a critical role in fighting a-gainst COVID-19.Although a large number of epitope-based and mucosal vaccines have been stud-ied,few peptide epitope vaccines targeting the mucosa and their functional evaluation have been re-ported.In this study,we used SARS-CoV-2 structural protein M peptide epitope predicted by the IEDB database as an antigenic target to design the MS-3S gene containing 3 050 and 1 229 signal peptides and DCpep optimized for insertion into MS2 phage coat proteins.The expression plasmid pSIP:MS-3S was constructed by cloning the PCR fragments seamlessly and was transformed into Lactiplantibacillus plantarum 18 to obtain the recombinant bacterium LP18:MS-3S.Expression conditions such as induction time,inducer concentration,rotational speed and initial pH were opti-mized.The intranasal immunization experiments were performed to examine the vaccine efficacy.The results showed that the 916 bp-long target gene MS-3S modified and optimized was amplified and used to successfully construct the recombinant bacterial strain LP18:MS-3S.The optimal con-ditions for recombinant protein expression were obtained and verified by Western blot,flow cy-tometry,immunofluorescence and other detection methods.The optimal expression conditions were determined as follows:induction time was 4 h with 100 pg/L of SppIP as the optimal induction concentration.Antibody-specific for the epitope was verified by ELISA experiments in serum,alve-olar lavage fluid and fecal dilutions of mice.In summary,a recombinant bacterial strain expressing the epitope antigen of the SARS-CoV-2 M protein peptide was constructed.The obtained protein can induce the body to produce humoral and mucosal immunity,which lays the foundation for the development of a vaccine candidate for the mucosal immunity of COVID-19.

Objective:To perform optical surface monitoring-based three-dimensional (3D) in vivo dose verification for patients with left breast cancer undergoing deep inspiration breath-hold surface-guided radiation therapy (DIBH-SGRT) and to investigate the dosimetric differences in the target volumes and related factors affecting γ pass rates. Methods:Totally 20 patients with left breast cancer who received DIBH-SGRT at the Department of Radiation Oncology, Women′s Hospital, School of Medicine, Zhejiang University were selected. The optical surface monitoring-based intrafractional displacement deviations of the patients during DIBH were recorded. Meanwhile, electronic portal imaging device (EPID)-based in vivo dosimetry (EIVD) verification was performed for patients during the DIBH-SGRT, and γ pass rates were measured with the criteria of 2 mm/2%, 3 mm/3%, and 3 mm/5%. The dosimetric differences between planning target volumes (PTVs) and organs at risk (OARs) were analyzed based on dose-volume histograms (DVHs). Furthermore, Pearson correlation analysis was employed to determine the correlation of three γ pass rates with dosimetric differences and displacement deviations. Results:The average pass rates with the criteria of 2 mm/2%, 3 mm/3%, and 3 mm/5% were determined at 73.43%, 86.00%, and 92.96%, respectively, and the average deviations between EIVD measured doses and planned doses in PTV_TB and PTV Dmean were proved to be 0.23% and 0.59%, respectively ( P > 0.05). Pearson analysis revealed that the γ pass rates exhibited a weak correlation with dosimetric differences in PTVs( R<0.7) but strong correlations with intrafractional displacement deviations in Lat and Vert directions during DIBH ( R > 0.7). Conclusions:EIVD verification can ensure the high accuracy of dose delivery in PTVs during DIBH-SGRT for left breast cancer. Additionally, the EIVD verification system has the potential to detect displacement deviations during breath holding.

Objective:To investigate the efficacy and safety of retroperitoneal soft tissue sarcoma resection combined with nephrectomy.Methods:The clinical data of 27 cases undergoing retroperitoneal soft tissue sarcoma resection combined with nephrectomy at the Gastrointestinal Tumor Center , Guangdong Provincial Hospital , Traditional Chinese Medicine from Jun 2017 to Aug 2022 were retrospectively analyzed for the indication of nephrectomy, postoperative progression of renal insufficiency and survival rate.Results:Twenty-six cases (96%) achieved R 0/R 1 resection and 1 case nderwent R 2 resection. Six cases underwent combined unilateral nephrectomy and 21 patients underwent combined multi-organ resection with a median number of resections of 4 (2,5). Postoperative pathology suggested that the combined resected kidney was positive for tumor infiltration in 17 cases. Five cases had Clavien-Dindo grade 3 or higher complications and no deaths occurred within 30 days after surgery. At the 90th day after surgery, 19 cases (70%) had decreased renal function ( Z=2.88, P=0.04), with a median decrease of -3.96 (-30.36, 0.31)ml·(min·1.73 m 2) -1, including 8 cases of preoperative Chronic Kidney Disease(CKD)1 stage progression (6 cases of CKD 2 stage, 2 cases of CKD 3 stage); 2 cases of CKD 2 stage progressed to CKD 3 stage; 1 case of preoperative CKD 3 stage progressed to CKD 4 stage. During the follow-up period of 3-38 months, no patient progressed to CKD 5 stage and no patient required dialysis treatment. Conclusion:Retroperitoneal soft tissue sarcoma resection combined with nephrectomy is safe and feasible while improving tumor radicality.

Excessive N-acetyl-p-benzoquinone imine(NAPQI)formation is a starting event that triggers oxidative stress and subsequent hepatocyte necrosis in acetaminophen(APAP)overdose caused acute liver failure(ALF).S-glutathionylation is a reversible redox post-translational modification and a prospective mechanism of APAP hepatotoxicity.Glutaredoxin-1(Glrx1),a glutathione-specific thioltransferase,is a primary enzyme to catalyze deglutathionylation.The objective of this study was to explored whether and how Glrx1 is associated with the development of ALF induced by APAP.The Glrx1 knockout mice(Glrx1-/-)and liver-specific overexpression of Glrx1(AAV8-Glrx1)mice were produced and underwent APAP-induced ALF.Pirfenidone(PFD),a potential inducer of Glrx1,was administrated preceding APAP to assess its protective effects.Our results revealed that the hepatic total protein S-glutathionylation(PSSG)increased and the Glrx1 level reduced in mice after APAP toxicity.Glrx1-/- mice were more sensitive to APAP overdose,with higher oxidative stress and more toxic metabolites of APAP.This was attributed to Glrx1 deficiency increasing the total hepatic PSSG and the S-glutathionylation of cytochrome p450 3a 11(Cyp3a11),which likely increased the activity of Cyp3a11.Conversely,AAV8-Glrx1 mice were defended against liver damage caused by APAP overdose by inhibiting the S-glutathionylation and activity of Cyp3a11,which reduced the toxic metabolites of APAP and oxidative stress.PFD precede administration upregulated Glrx1 expression and alleviated APAP-induced ALF by decreasing oxidative stress.We have identified the function of Glrx1 mediated PSSG in liver injury caused by APAP overdose.Increasing Glrx1 expression may be investigated for the medical treatment of APAP-caused hepatic injury.