OBJECTIVE To investigate the effects and mechanisms of Lycium ruthenicum Murr. in improving sleep. METHODS Network pharmacology was employed to identify the active components of L. ruthenicum and their associated disease targets， followed by enrichment analysis. A caffeine‑induced zebrafish model of sleep deprivation was established ， and the zebrafish were treated with L. ruthenicum Murr. extract （LRME） at concentrations of 0.1， 0.2 and 0.4 mg/mL， respectively； 24 h later， behavioral changes of zebrafish and pathological alterations in brain neurons were subsequently observed. The levels of inflammatory factors [interleukin-6 （IL-6）， IL-1β， IL-10， tumor necrosis factor-α （TNF-α）]， oxidative stress markers [superoxide dismutase （SOD）， malondialdehyde （MDA）， glutathione peroxidase （GSH-Px）， catalase （CAT）]， and neurotransmitters [5- hydroxytryptamine （5-HT）， γ-aminobutyric acid （GABA）， glutamic acid （Glu）， dopamine （DA）， and norepinephrine （NE）] were measured. The protein expression levels of protein kinase B1 （AKT1）， phosphorylated AKT1 （p-AKT1）， epidermal growth factor receptor （EGFR）， B-cell lymphoma 2 （Bcl-2）， sarcoma proto-oncogene，non-receptor tyrosine kinase （SRC）， and heat shock protein 90α family class A member 1 （HSP90AA1） in the zebrafish were also determined. RESULTS A total of 12 active components and 176 intersecting disease targets were identified through network pharmacology analysis. Among these， apigenin， naringenin and others were recognized as core active compounds， while AKT1， EGFR and others served as key targets； EGFR tyrosine kinase inhibitor resistance signaling pathway was identified as the critical pathway. The sleep improvement rates in zebrafish of LRME low-， medium-， and high-dose groups were 54.60%， 69.03% and 77.97%， 开发。E-mail：hjp_yft@163.com respectively， while the inhibition ratios of locomotor distance were 0.57， 0.83 and 0.95， respectively. Compared with the model group， the number of resting counts， resting time and resting distance were significantly increased/extended in LRME medium- and high-dose groups （P＜0.05）. Neuronal damage in the brain was alleviated. Additionally， the levels of IL-6， IL-1β， TNF-α， MDA， Glu， DA and NE， as well as the protein expression levels of AKT1， p-AKT1， EGFR， SRC and HSP90AA1， were markedly reduced （P＜0.05）， while the levels of IL-10， SOD， GSH-Px， CAT， 5-HT and GABA， as well as Bcl-2 protein expression， were significantly elevated （P＜0.05）. CONCLUSIONS L. ruthenicum Murr. demonstrates sleep-improving effects， and its specific mechanism may be related to the regulation of inflammatory responses， oxidative stress， neurotransmitter balance， and the EGFR tyrosine kinase inhibitor resistance signaling pathway.

ObjectiveTo clarify whether METTL14 mediates the core role of acupuncture at Neiguan (PC6) in promoting myelination and improving behavior in young autistic rats through gene intervention technology. MethodsThe ASD model was established by intraperitoneal injection of valproic acid (VPA) in pregnant rats. Male offspring were intracerebroventricularly injected with adenovirus-packaged METTL14 shRNA (sh-METTL14) or its control (sh-NC) on postnatal day 1, with a model group set as well. Subsequently, the juvenile rats were divided into model group, acupuncture group, acupuncture+sh-NC group, and acupuncture+sh-METTL14 group. The acupuncture group received acupuncture at Neiguan (PC6) from postnatal day 7, once daily for 21 consecutive days. Neurobehavioral changes were evaluated by behavioral tests; METTL14 knockdown efficiency and the expression of METTL14, METTL3, and PTEN were detected by quantitative real-time PCR (qRT-PCR) and Western blot (WB); PTEN m⁶A levels were measured by RNA immunoprecipitation-qPCR (RIP-qPCR); myelin ultrastructure, expression of myelin basic protein (MBP) and neurofascin 155 (NF155), and dendritic spine density were observed using transmission electron microscopy (TEM), enzyme-linked immunosorbent assay (ELISA), immunofluorescence, qRT-PCR, and primary neuron culture. ResultsBehaviorally, knockdown of METTL14 significantly counteracted the beneficial effects of acupuncture in improving self-grooming, open field exploration, three-chamber social interaction, and Morris water maze learning and memory (P<0.05, P<0.01). Compared with the acupuncture+sh-NC group, the acupuncture+sh-METTL14 group showed significantly decreased mRNA and protein expression of hippocampal METTL14 (P<0.01), and the upregulating effects of acupuncture on METTL3 and PTEN expression were reversed (P<0.01). Meanwhile, knockdown of METTL14 significantly inhibited the acupuncture-induced increase in PTEN m⁶A levels (P<0.01). Morphologically, knockdown of METTL14 attenuated the improvement of myelin structure by acupuncture, reversed the downregulation of MBP and upregulation of NF155 induced by acupuncture, and blocked the increase in dendritic spine density (P<0.05, P<0.01). ConclusionMETTL14 is a key molecule mediating the therapeutic effect of acupuncture at Neiguan. Acupuncture at Neiguan upregulates METTL14, thereby enhancing m⁶A methylation modification of PTEN mRNA to stabilize its expression, ultimately promoting myelin development and improving behavioral symptoms in ASD juvenile rats. This preliminarily reveals the modern biological connotation of “opening Xuanfu and dredging myelin”.

ObjectiveTo clarify whether METTL14 mediates the core role of acupuncture at Neiguan (PC6) in promoting myelination and improving behavior in young autistic rats through gene intervention technology. MethodsThe ASD model was established by intraperitoneal injection of valproic acid (VPA) in pregnant rats. Male offspring were intracerebroventricularly injected with adenovirus-packaged METTL14 shRNA (sh-METTL14) or its control (sh-NC) on postnatal day 1, with a model group set as well. Subsequently, the juvenile rats were divided into model group, acupuncture group, acupuncture+sh-NC group, and acupuncture+sh-METTL14 group. The acupuncture group received acupuncture at Neiguan (PC6) from postnatal day 7, once daily for 21 consecutive days. Neurobehavioral changes were evaluated by behavioral tests; METTL14 knockdown efficiency and the expression of METTL14, METTL3, and PTEN were detected by quantitative real-time PCR (qRT-PCR) and Western blot (WB); PTEN m⁶A levels were measured by RNA immunoprecipitation-qPCR (RIP-qPCR); myelin ultrastructure, expression of myelin basic protein (MBP) and neurofascin 155 (NF155), and dendritic spine density were observed using transmission electron microscopy (TEM), enzyme-linked immunosorbent assay (ELISA), immunofluorescence, qRT-PCR, and primary neuron culture. ResultsBehaviorally, knockdown of METTL14 significantly counteracted the beneficial effects of acupuncture in improving self-grooming, open field exploration, three-chamber social interaction, and Morris water maze learning and memory (P<0.05, P<0.01). Compared with the acupuncture+sh-NC group, the acupuncture+sh-METTL14 group showed significantly decreased mRNA and protein expression of hippocampal METTL14 (P<0.01), and the upregulating effects of acupuncture on METTL3 and PTEN expression were reversed (P<0.01). Meanwhile, knockdown of METTL14 significantly inhibited the acupuncture-induced increase in PTEN m⁶A levels (P<0.01). Morphologically, knockdown of METTL14 attenuated the improvement of myelin structure by acupuncture, reversed the downregulation of MBP and upregulation of NF155 induced by acupuncture, and blocked the increase in dendritic spine density (P<0.05, P<0.01). ConclusionMETTL14 is a key molecule mediating the therapeutic effect of acupuncture at Neiguan. Acupuncture at Neiguan upregulates METTL14, thereby enhancing m⁶A methylation modification of PTEN mRNA to stabilize its expression, ultimately promoting myelin development and improving behavioral symptoms in ASD juvenile rats. This preliminarily reveals the modern biological connotation of “opening Xuanfu and dredging myelin”.

Objective:To investigate the current status of vascular access team building and analysis its related factors in hemodialysis centers in China.Methods:The study was a cross-sectional survey. Using a convenience sampling method, a questionnaire was designed to investigate the clinical practice of vascular access teams in 527 hemodialysis centers in China from March to April 2022. The related factors of the formation of vascular access teams and the setting up of vascular access coordinators (VAC) were analyzed by multivariate logistic regression method.Results:A total of 506 valid questionnaires were recovered, with a recovery rate of 96.02%. There were 247 (48.81%) and 193 (38.14%) hemodialysis centers respectively across China that had built vascular access teams and set up VAC. Hemodialysis centers with more than 10 years of practice had higher rate of implementation than those in hemodialysis centers with practice years less or equal than 10 years in developing standardized procedures for vascular access management ( χ 2=8.288, P=0.004), holding continuous quality improvement meetings on vascular access ( χ 2=8.210, P=0.004), establishing vascular access teams ( χ 2=33.805, P<0.001) and setting up vascular access coordinators ( χ 2=16.038, P<0.001), and the difference was statistically significant. The results of multivariate logistic regression analysis showed that the number of dialysis machines ( OR=2.221, 95% CI 1.118-4.415, P=0.023), the number of patients on dialysis( OR=2.946, 95% CI 1.375-6.310, P=0.005), and the establishment of VAC positions ( OR=9.463, 95% CI 5.307-16.874, P<0.001), and the standardized vascular access management process ( OR=3.383, 95% CI 2.012-5.687, P<0.001) were the related factors of vascular access team building. The related factors of setting up a VAC position in hemodialysis center were opening vascular access clinic ( OR=2.704,95% CI 1.382-5.290, P=0.004), the formation of a vascular access team ( OR=9.464, 95% CI 5.312-16.860, P<0.001), and constructing standardized procedures for vascular access management ( OR=3.663, 95% CI 2.243-5.982, P<0.001). Conclusion:The implementation rates of vascular access team and VAC position in hemodialysis centers were 48.81% and 38.14%, respectively. The number of dialysis machines, the number of patients on dialysis, the standardized procedures for vascular access management, the vascular access clinic, the vascular access team, and the VAC position were the relevant factors of the team building for vascular access.

Objective The aim of this study is to examine the Chinese version of Visual Prostate Symptom Score(CVPSS)and the International Prostate Symptom Score(IPSS)in the assessment of lower urinary tract symptoms in BPH pa-tients.Methods By using convenient sampling,inpatients in the urology department of a tertiary grade A hospital in Shanghai were selected as the survey subjects from March 2023 to March 2024.The lower urinary tract symptoms of the patients were eval-uated using the self-designed general information questionnaire.And the CVPSS and IPSS with their urine flow rate were meas-ured.A comparative analysis was conducted from aspects such as internal consistency,correlation of item scores,completion time of the scale,and assistance rate.Results A total of 246 patients with BPH were recruited.The total score and life quality score were 13.93±3.55 and 4.23±1.02 by CVPSS.And the total score and life quality by IPSS was 18.33±7.55 and 4.36±1.02,respectively.The Cronbach's α coefficient were 0.761 and 0.787,respectively.The time taken on CVPSS was less than that on IPSS(P＜0.01).And the rate of needing assistance was 23.58％for CVPSS,which was significantly lower than that(65.24％)for IPSS.Conclusion CVPSS is significantly correlated with the corresponding items and total scores of IPSS,as well as the quality of life.Moreover,it takes less time and can be used as a simple and effective self-assessment tool for lower urinary tract symptoms in elderly BPH patients with lower education levels.It reduces the burden of medical staffs as well.

Objective:To evaluate the value of ultrasound radiomics combined with machine learning for early diagnosis of seronegative Hashimoto’s thyroiditis (SN-HT) .Methods:This retrospective study included 164 patients from Liaoning Provincial People’s Hospital , Lixin County People’s Hospital, Linghai Dalinghe Hospital, Fengcheng Phoenix Hospital, who underwent thyroidectomy for solitary nodules with normal thyroid function between Nov. 2016 and Jan. 2024. Postoperative pathology confirmed Hashimoto’s thyroiditis (HT) in some cases, who were further categorized into antibody-positive and antibody-negative groups based on serum antibody status. Patients without Hashimoto’s thyroiditis served as the control group. A total of 298 ultrasound images were analyzed. Radiomics features were extracted from hypoechoic non-nodular areas within 0.5 cm surrounding the tumor. Two senior pathologists and two senior ultrasound physicians independently assessed lymphocytic infiltration, eosinophilic changes of follicular epithelium, and the proportion of hypoechoic areas in pathology and ultrasound images, respectively. A machine learning model, CCH-NET, was developed using linear regression and t-distributed stochastic neighbor embedding (t-SNE) techniques. The dataset was divided into a training set (80%) and a validation set (20%) to compare the diagnostic accuracy of CCH-NET with that of senior ultrasound physicians. Results:In internal validation, CCH-NET achieved a diagnostic accuracy of 88.89% for both antibody-positive and antibody-negative groups, significantly higher than the 66.67% accuracy of senior ultrasound physicians ( P<0.01). In external validation, CCH-NET achieved 75.00% and 66.67% accuracy for the two groups, compared to 50.00% by senior ultrasound physicians. For the control group, both methods achieved 93.33% accuracy. The AUC of CCH-NET was 0.848, outperforming senior ultrasound physicians (0.681) ,demonstrating superior diagnostic performance. Conclusion:The radiomics-based CCH-NET model, using non-nodular hypoechoic areas as a specific indicator, can accurately identify early SN-HT in euthyroid patients. It significantly outperforms senior ultrasound physicians, improving diagnostic accuracy and reducing missed diagnoses.

Objective The aim of this study is to examine the Chinese version of Visual Prostate Symptom Score(CVPSS)and the International Prostate Symptom Score(IPSS)in the assessment of lower urinary tract symptoms in BPH pa-tients.Methods By using convenient sampling,inpatients in the urology department of a tertiary grade A hospital in Shanghai were selected as the survey subjects from March 2023 to March 2024.The lower urinary tract symptoms of the patients were eval-uated using the self-designed general information questionnaire.And the CVPSS and IPSS with their urine flow rate were meas-ured.A comparative analysis was conducted from aspects such as internal consistency,correlation of item scores,completion time of the scale,and assistance rate.Results A total of 246 patients with BPH were recruited.The total score and life quality score were 13.93±3.55 and 4.23±1.02 by CVPSS.And the total score and life quality by IPSS was 18.33±7.55 and 4.36±1.02,respectively.The Cronbach's α coefficient were 0.761 and 0.787,respectively.The time taken on CVPSS was less than that on IPSS(P＜0.01).And the rate of needing assistance was 23.58％for CVPSS,which was significantly lower than that(65.24％)for IPSS.Conclusion CVPSS is significantly correlated with the corresponding items and total scores of IPSS,as well as the quality of life.Moreover,it takes less time and can be used as a simple and effective self-assessment tool for lower urinary tract symptoms in elderly BPH patients with lower education levels.It reduces the burden of medical staffs as well.

Objective:To evaluate the value of ultrasound radiomics combined with machine learning for early diagnosis of seronegative Hashimoto’s thyroiditis (SN-HT) .Methods:This retrospective study included 164 patients from Liaoning Provincial People’s Hospital , Lixin County People’s Hospital, Linghai Dalinghe Hospital, Fengcheng Phoenix Hospital, who underwent thyroidectomy for solitary nodules with normal thyroid function between Nov. 2016 and Jan. 2024. Postoperative pathology confirmed Hashimoto’s thyroiditis (HT) in some cases, who were further categorized into antibody-positive and antibody-negative groups based on serum antibody status. Patients without Hashimoto’s thyroiditis served as the control group. A total of 298 ultrasound images were analyzed. Radiomics features were extracted from hypoechoic non-nodular areas within 0.5 cm surrounding the tumor. Two senior pathologists and two senior ultrasound physicians independently assessed lymphocytic infiltration, eosinophilic changes of follicular epithelium, and the proportion of hypoechoic areas in pathology and ultrasound images, respectively. A machine learning model, CCH-NET, was developed using linear regression and t-distributed stochastic neighbor embedding (t-SNE) techniques. The dataset was divided into a training set (80%) and a validation set (20%) to compare the diagnostic accuracy of CCH-NET with that of senior ultrasound physicians. Results:In internal validation, CCH-NET achieved a diagnostic accuracy of 88.89% for both antibody-positive and antibody-negative groups, significantly higher than the 66.67% accuracy of senior ultrasound physicians ( P<0.01). In external validation, CCH-NET achieved 75.00% and 66.67% accuracy for the two groups, compared to 50.00% by senior ultrasound physicians. For the control group, both methods achieved 93.33% accuracy. The AUC of CCH-NET was 0.848, outperforming senior ultrasound physicians (0.681) ,demonstrating superior diagnostic performance. Conclusion:The radiomics-based CCH-NET model, using non-nodular hypoechoic areas as a specific indicator, can accurately identify early SN-HT in euthyroid patients. It significantly outperforms senior ultrasound physicians, improving diagnostic accuracy and reducing missed diagnoses.

Objective:To investigate the current status of vascular access team building and analysis its related factors in hemodialysis centers in China.Methods:The study was a cross-sectional survey. Using a convenience sampling method, a questionnaire was designed to investigate the clinical practice of vascular access teams in 527 hemodialysis centers in China from March to April 2022. The related factors of the formation of vascular access teams and the setting up of vascular access coordinators (VAC) were analyzed by multivariate logistic regression method.Results:A total of 506 valid questionnaires were recovered, with a recovery rate of 96.02%. There were 247 (48.81%) and 193 (38.14%) hemodialysis centers respectively across China that had built vascular access teams and set up VAC. Hemodialysis centers with more than 10 years of practice had higher rate of implementation than those in hemodialysis centers with practice years less or equal than 10 years in developing standardized procedures for vascular access management ( χ 2=8.288, P=0.004), holding continuous quality improvement meetings on vascular access ( χ 2=8.210, P=0.004), establishing vascular access teams ( χ 2=33.805, P<0.001) and setting up vascular access coordinators ( χ 2=16.038, P<0.001), and the difference was statistically significant. The results of multivariate logistic regression analysis showed that the number of dialysis machines ( OR=2.221, 95% CI 1.118-4.415, P=0.023), the number of patients on dialysis( OR=2.946, 95% CI 1.375-6.310, P=0.005), and the establishment of VAC positions ( OR=9.463, 95% CI 5.307-16.874, P<0.001), and the standardized vascular access management process ( OR=3.383, 95% CI 2.012-5.687, P<0.001) were the related factors of vascular access team building. The related factors of setting up a VAC position in hemodialysis center were opening vascular access clinic ( OR=2.704,95% CI 1.382-5.290, P=0.004), the formation of a vascular access team ( OR=9.464, 95% CI 5.312-16.860, P<0.001), and constructing standardized procedures for vascular access management ( OR=3.663, 95% CI 2.243-5.982, P<0.001). Conclusion:The implementation rates of vascular access team and VAC position in hemodialysis centers were 48.81% and 38.14%, respectively. The number of dialysis machines, the number of patients on dialysis, the standardized procedures for vascular access management, the vascular access clinic, the vascular access team, and the VAC position were the relevant factors of the team building for vascular access.

Objective To explore the toxicity of Polygonum multiflorum alcohol extract on 3D hepatospheres.Methods Variations in culture conditions and cell ratios were implemented,followed by the assessment of cell sphere diameter,density,and roundness,aiming to explore the optimal culture conditions.The 3D hepatocyte spheres were divided into control group and experimental-L,-M,-H groups.The experimental-L,-M,-H groups were treated with 0.25,1.00 and 2.50 mg·mL-1 Polygounm multiforum alcohol extract,and the control group was given the same amount of culture medium.The cell viability of the cell spheroids was tested by CellTiter-Glo reagent,the expression level of liver function related genes was detected by fluorescent quantitative polymerase chain reaction(RT-qRCR).The toxicity of cell spheres was detected by double fluorescent staining of living and dead cells.Results The ideal culture condition of cell sphere was 500 cells per micropore,and the cell ratio was HepG2-Huvec-LX-2=8∶1∶1.It displayed the values of 0.91±0.07 for circularity,0.91±0.02 for firmness,1.12±0.14 for aspect ratio,and(170.97±14.79)μm for diameter.On the 3rd,7th,10th and 14th days,the expression levels of albumin(ALB)mRNA were 1.00±0.02,0.96±0.02,0.54±0.07,0.52±0.07,and the expression levels of cytochrome P450 1A2(CYP1A2)mRNA were 1.00±0.10,2.15±0.16,2.45±0.33,1.30±0.03,respectively.The expression levels of multidrug resistance protein 2(MPR2)in the control group and the experimental-L,-M,-H groups were 1.00±0.31,1.38±0.24,1.48±0.06 and 1.90±0.08,respectively;spheroid viability were(98.19±0.49)％,(88.53±0.90)％,(71.60±2.91)％and(56.65±5.41)％.There were statistically significant differences in the above indexes between the experimental-L,-M,-H groups and the control group(all P＜0.05).Conclusion The established hepatocyte sphere co-culture model showed varying degrees of expression of phase Ⅰ/Ⅱ drug metabolism enzymes,transporters,and liver cell specific marker molecule albumin and can be used to evaluate the toxicity of multiflorum multiflorum,which provides further reference for the clinical application of multiflorum multiflorum.