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.

OBJECTIVE To investigate the effects of total flavonoids from Carthamus tinctorius L. （TFCTL） on hepatic stellate cell （HSC） activation based on the microRNA （miRNA）-204/NUAK family SNF1-like kinase 1 （NUAK1）/Hippo signaling axis， thereby elucidating the potential mechanism underlying their antifibrotic effects. METHODS The HSC-T6 cells were divided into control group， model group， TFCTL low-concentration group （20 μg/mL）， TFCTL medium-concentration group （40 μg/mL）， and TFCTL high-concentration group （60 μg/mL）. Except for control group， the remaining groups were treated with 5 ng/mL of transforming growth factor-β to induce the activation of hepatic stellate cells， followed by the addition of corresponding drug solutions/culture medium and incubation for 24 hours. Cell apoptosis was assessed， the expression levels of α-smooth muscle actin （α-SMA）， type Ⅰ collagen （Collagen Ⅰ） and proteins associated with the Hippo/Yes-associated protein （YAP） pathway [YAP， large tumor suppressor kinase 1 （LATS1）， and mammalian STE20-like kinase 1 （MST1）] were detected. Additionally， cell transfection was used to investigate the activity of the miRNA-204/NUAK1/Hippo signaling axis at both the genetic and protein levels. RESULTS After intervention with TFCTL， the apoptosis rate of HSC-T6 cells and the protein expressions of MST1 （except for the TFCTL high-concentration group） and LATS1 were significantly increased （P＜0.05）， while the protein expressions of α-SMA， CollagenⅠ， and YAP （except for the TFCTL medium-concentration group） were significantly decreased （P＜0.05）. Further results from cell transfection experiments revealed that after transfection with miRNA-204 mimics， the mRNA it’s protein expressions of α-SMA， CollagenⅠ， NUAK1， and YAP in HSC-T6 cells were significantly decreased （P＜0.05）， while the mRNA and protein expressions of LATS1 and the mRNA expression of MST1 were significantly increased （P＜0.05）. Conversely， the results were opposite following transfection with miRNA-204 inhibitors. CONCLUSIONS TFCTL can exert anti-hepatic fibrosis effects by up-regulating the expression of miRNA-204， thereby down- regulating the expressions of NUAK1， inactivating the Hippo/YAP pathway， which in turn suppresses the activation of HSC and promotes their apoptosis.

Objective To explore the diagnostic value of mediastinal lymph nodes CT image features in pulmonary sarcoidosis.Methods The imageologic data of the patients with pulmonary sarcoidosis(pulmonary sarcoidosis group)and lung cancer complicating mediastinal lymph node metastasis(lung cancer group)con-firmed by EBUS-TBNA in this hospital from June 2015 to November 2023 were analyzed retrospectively.The chest plain scan CT and enhanced scan CT were performed in all cases,and the imaging characteristics were compared between the two groups.The receiver operating characteristic(ROC)curve was drawn and the area under the curve(AUC)was calculated.The diagnostic value of CT imaging characteristics in pulmonary sar-coidosis was analyzed.Results The proportion of female patients in the lung sarcoidosis group was signifi-cantly higher than that in the lung cancer group,while the age was smaller than that in the lung cancer group,and the differences were statistically significant(P＜0.05).The number of mediastinal lymph nodes,location,short diameter and CT enhancement value in the pulmonary sarcoidosis group had statistical difference(P＜0.05).The ROC curve analysis showed that AUC of lymph node short diameter reciprocal,CT enhancement value and position in detection alone were 0.586,0.785 and 0.505 respectively,and AUC of the three combined reciprocal was the highest(0.789).Conclusion The lymph node short diameter,CT enhancement value and po-sition have certain value in clinical diagnosis of pulmonary sarcoidosis and the 3-indicator combination could increase the diagnostic efficiency of pulmonary sarcoidosis.

Objective:To explore the application of contrast-enhanced ultrasound in the preoperative evaluation of carotid body tumor (CBT).Methods:The clinical data of 13 CBT patients undergoing contrast enhanced ultrasound test and surgical treatment at Peking Union Medical College Hospital from Nov 2017 to Aug 2021 was retrospectively analyzed.Results:Among the 13 patients, 7 patients had bilateral lesions. 18 tumors were identified by contrast enhanced ultrasound, which showed rich blood supply, with marked enhancement in 13 tumors and moderate enhancement in 5 tumors. The origins of the arterial supply for tumors were identified by contrast enhanced ultrasound. Time-intensity curve analysis showed that the tumors had enhancement characteristics of fast wash in and slow wash out. The mean contrast wash in time was (3.33±1.40) s, the mean peak intensity was (10.41±1.74) dB, and the mean wash out time was (56.47±22.28) s. A total of 13 cases underwent successful surgical removal. Five cases of external carotid artery ligation and 2 cases of internal carotid artery reconstruction were performed during surgery. Postoperative transient neurological injury occurred in 5 cases. There were no cases of cerebral infarction or death in the perioperative period. Mean postoperative follow-up was 14.31 months. Five cases of neurological injury had satisfactory recovery and no other adverse events occurred.Conclusions:Contrast enhanced ultrasound is an effective method of preoperative imaging assessment for CBT, which helps the surgical planning and preoperative preparation.

Objective To develop nanobodies with broad-spectrum reactivity,specificity,and high sensitivity that can be used for detecting multiple subtypes of influenza A virus,and to establish a nanobody-based enzyme-linked immunosorbent assay(ELISA)method.Methods Gene sequences of twelve nanobodies against influenza A virus were retrieved from the National Center for Biotechnology Information(NCBI)and nanobody databases.The nanoantibodies were prepared using molecular biological techniques including gene synthesis and recombinant expression.The binding activity,specificity,sensitivity,and affinity of these nanobodies were determined by ELISA screening and Gator affinity analysis.A double-antibody sandwich ELISA assay was established by combining the selected nanobody with a traditional mouse monoclonal antibody.Results Twelve nanobodies were expressed and purified.Two nanobodies capable of binding to multiple subtypes of influenza virus including H1,H3,H5,H7,and H9 were obtained and designated as VHH54 and KV108.Both nanobodies showed no cross-reactivity with other respiratory virus antigens.Furthermore,the KV108 nanobody exhibited the highest binding affinity,with a dissociation constant of 5.94×10-9mol/L for the influenza virus nucleoprotein(NP),and the lowest detection concentration for the NP antigen reached 0.00064 μg/mL.The double-antibody sandwich ELISA,using a combination of KV108 and a mouse monoclonal antibody,could sensitively detect the five common subtypes of influenza A virus(H1N1,H3N2,H5N1,H7N9,and H9N2).The lowest detection limit reached 110-403 PFU/mL,which was higher than that of the commercial colloidal gold kitfor influenza virus detection.Conclusion This study has identified a nanobody KV108,which is capable of binding to multiple subtypes of influenza virus,and established a nanobody-based ELISA method that can detect multiple subtypes of influenza A virus.This study can facilitate the development of nanobody-based influenza detection technologies.

Objective To investigate the application effect of a refined early diet plan in patients with distal gastric cancer after laparoscopic radical surgery.Methods By convenience sampling method,46 patients with laparoscopic radical gastrectomy for distal gastric cancer admitted to a tertiary hospital in Zhejiang Province from July to December 2023 were selected as an experimental group,while 49 patients admitted from January to June 2023 as a control group.The experimental group was administered with the refined early diet plan,and the control group was administered with conventional methods.The intervention period was from the first day after surgery to discharge.The differences of the first postoperative defecation time,perioperative thirst and hunger score,postoperative hospitalization time,hospitalization cost,postoperative complications and the incidence of readmission 30 days after surgery were compared between the 2 groups.Results 42 patients in the experimental group and 49 patients in the control group completed the study.First postoperative exhaust time(t=4.922,P＜0.001),first postopera-tive defecation time(Z=-2.440,P=0.015),perioperative thirst score(Z=-8.024,P＜0.001),perioperative hunger score(Z=-8.192,P＜0.00 1),postoperative hospitalization time(Z=-7.622,P＜0.001)and hospitalization cost(Z=-4.522,P＜0.001)were lower than those of the control group,and the difference was statistically significant.There was no significant difference in complication rate and 30-day readmission rate between the experimental group and the control group(P＞0.05).Conclusion The refined early diet plan is safe and effective for early recovery of patients after laparoscopic radical gastrectomy for distal gastric cancer,promoting intestinal function recovery,shortening hospital stay,reducing hospital costs,and improving patients'subjective comfort.

Hepatocellular carcinoma(HCC)is one of the leading causes of cancer-related mortality worldwide.As a typical immunogenic tumor,HCC has a complex immune microenvironment that plays a crucial role in its initiation and progression.In recent years,the interactive network between immunosuppressive cells and circulating tumor cells(CTCs)has drawn increasing attention for its role in promoting HCC immune evasion and metastasis.However,how CTCs escape immune surveillance and clearance by immune cells remains unclear.This article focuses on the regulatory networks between CTCs and immunosuppressive cells as well as the key pathways of immune evasion,introduces exosome-mediated regulation and single-cell multi-omics technologies to dissect mechanisms of cellular interactions,systematically discusses the clinical translational challenges of targeting pathways such as PD-L1 and CCL5,and proposes future prospects for personalized therapy driven by artificial intelligence.

OBJECTIVE: To observe the effect of the direct stimulation of acupuncture at extraocular muscle attachment point on acquired extraocular muscle palsy. METHODS: Thirteen patients with acquired extraocular muscle palsy were treated with acupuncture directly at extraocular muscle (paralytic muscle) attachment point. Firstly, the intraocular conjunctival sac drops of topical anesthetic (procaine hydrochloride eye drops) were administered, 0.2 mL each time, once every 10 minutes, for a total of 3 times. Acupuncture was delivered immediately after the third drop. The sterile acupuncture needle for single use, 0.25 mm×25 mm, was inserted at the anatomical location of the corneal limbal attachment of paralytic extraocular muscle, with an angle of 10° to 15° formed between the needle tip and extraocular muscle, and a depth of 0.3 mm to 0.5 mm. Pivoted by the needle tip, the eyeball was moved passively towards the direction of normal action of orbital muscle, 30 to 50 times until the patient felt soreness of the eyeball; afterwards, the needle was removed. After acupuncture, levofloxacin eye drops were administered once (0.2 mL) at the affected eye. The treatment was given twice a week, and completed when diplopia disappeared. Before and after treatment, the diplopia and the synoptophore circumference were observed respectively. RESULTS: After 7 to 24 (15.46±5.56) times of direct stimulation with acupuncture at extraocular muscle attachment point, the symptoms of diplopia disappeared in 13 patients, the eye position restored to orthophoria, and the circumference of synoptophore was reduced to be (4.04±0.82)° from (19.38±3.98)° detected before treatment (P<0.05). CONCLUSION Acupuncture directly at extraocular muscle attachment can attenuate diplopia and improve ocular muscle function in patients with acquired extraocular muscle palsy.
Humans ; Acupuncture Therapy ; Male ; Female ; Middle Aged ; Adult ; Oculomotor Muscles/physiopathology* ; Aged ; Acupuncture Points ; Ophthalmoplegia/physiopathology*

Objective To develop a virtual reality rehabilitation support system and evaluate its effects on improving discharge readiness in caregivers of laryngectomy patients.Methods The system is equipped with a virtual reality-based home nursing skills guidance module for post-laryngectomy patients and an evidence-based artificial intelligence Q&A module.A convenience sampling method was adopted to select 64 caregivers of patients who underwent laryngectomy for the first time in the otorhinolaryngology head and neck surgery department of a tertiary A hospital in Beijing from July 2024 to January 2025 for a systematic study on application effects.To avoid cross-contamination between an experimental group and a control group,ward-based grouping was applied.A coin toss was used to assign Ward 1 to an experimental group and Ward 2 to a control group,with 32 cases in each group.The experimental group received discharge guidance based on the virtual reality technology,while the control group was provided with conventional verbal and written discharge instructions.The readiness for hospital discharge and the quality of discharge teaching scores of caregivers were compared between the 2 groups after the intervention.Results No sample detachment.After the intervention,the patient caregivers in the experimental group scored significantly higher than those in the control group in terms of discharge readiness and quality of discharge guidance,with a statistically significant difference(P＜0.001).Conclusion This study developed the virtual reality rehabilitation system for laryngeal cancer patients demonstrated good feasibility and effectiveness.The application of this system for discharge education significantly improved discharge readiness of caregivers and quality of discharge guidance for post-laryngectomy patients.It also helped with the optimization of health education models,enhancement of nursing resource utilization efficiency,and improvement of transitional care.

Objective To investigate the effects of rutin on the proliferation and osteogenic differentiation of stem cells from human exfoliated deciduous teeth(SHEDs)and its regulatory relationship with the Janus protein tyrosine kinase 2(JAK2)/signal transducer and activator transcription 3(STAT3)pathway.Methods SHEDs were isolated and cultured in vitro.After identification,SHEDs were randomly grouped into the control group(normal culture),low,medium,and high dose rutin groups(0.1,0.5,1 μmol/L),and the inhibi-tor group(1 μmol/L of rutin+10 μmol/L of the JAK2/STAT3 pathway inhibitor AG490).Alizarin red staining was applied to detect the number of mineralized nodules,and the MTT method was utilized to detect cell proliferation.A commercial kit was used to detect cell alkaline phosphatase(ALP)activity,and quantitative real-time PCR was applied to detect the mRNA expression of Runt-related transcrip-tion factor 2(Runx2),osteocalcin(OCN),and alkaline phosphatase(ALP).Western blotting was used to detect the expression of Runx2,OCN,ALP,and JAK2/STAT3 signaling pathway proteins.Results Compared to that of the control group,the optical density value of SHEDs cells,the number of mineralized nodules,ALP activity,Runx2,OCN,ALP mRNA and protein levels,and p-JAK2/JAK2 p-STAT3/STAT3 were all markedly increased in the low,medium,and high dose rutin groups(P＜0.05).Treatment with the JAK2/STAT3 pathway inhibitor AG490 attenuated the enhancement of rutin on the proliferation and osteogenic differentiation of SHEDs(P＜0.05).Conclu-sion Rutin promotes the proliferation and osteogenic differentiation of SHEDs,and its mechanism is related to the activation of the JAK2/STAT3 signaling pathway.