ObjectiveTo analyze the pharmacodynamic activity of Bupleuri Radix processed with Trionyx sinensis blood in the treatment of Yin deficiency and study the spectrum-effect relationship of this medicine. MethodsHigh performance liquid chromatography was employed to establish the fingerprints of 15 batches of Bupleuri Radix processed with Trionyx sinensis blood， and the similarity was evaluated according to the SOP of Similarity Evaluation System of Chromatographic Fingerprint of TCM （version 2012）. A mouse model of Yin deficiency induced by thyroxine was established. The relationship between the active components and the effect on Yin deficiency was explored by grey correlation analysis and partial least squares method based on the changes in the serum levels of triiodothyronine （T3）， thyroxine （T4）， cyclic adenosine phosphate （cAMP）， and cyclic guanosine phosphate （cGMP）. The components screened out based on the spectrum-effect relationship were used for retrieval of the targets from the Traditional Chinese Medicine Systems Pharmacology and Analysis Database （TCMSP）， The Encyclopedia of Traditional Chinese Medicine （ETCM）， and Integrative Pharmacology-based Research Platform of Traditional Chinese Medicine （TCMIP）. Furthermore， the Online Mendelian Inheritance in Man （OMIM）， GeneCards， TTD， DisGeNET， and Drugbank were employed to establish the active component-target against Yin deficiency network of Bupleuri Radix processed with Trionyx sinensis blood. Gene Ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analyses were carried out for the core targets. Real-time PCR was conducted to verify the predicted key pathways and mechanisms. ResultsThe fingerprints of the 15 batches of Bupleuri Radix processed with Trionyx sinensis blood showed the similarities of 0.976-0.999 with the control fingerprint. Compared with the model group， the drug administration group showed elevated levels of T3 and T4 and lowered levels of cAMP， cGMP and cAMP/cGMP. The results of grey correlation analysis showed that active components in terms of the correlations followed the trend of saikosaponin B₁ > saikosaponin B₂ > saikosaponin C > saikosaponin D > saikosaponin A. The partial least squares analysis showed that saikosaponins A， D， B₁， and B₂ had higher VIP values. Network pharmacology predicted a total of 30 common targets， which were enriched in 276 GO terns and 115 KEGG pathways. The results of Real-time PCR showed that the model group had lower mRNA levels of Caspase-9， kinase insert domain receptor （KDR）， and mammalian target of rapamycin （mTOR） and higher mRNA level of mouse double minute 2 homolog （MDM2） than the blank group and the drug administration group. ConclusionBupleuri Radix processed with Trionyx sinensis blood has therapeutic effect on Yin deficiency syndrome， which provides a new idea for studying Bupleuri Radix processed with Trionyx sinensis blood.

Recent progress in targeted metabolic therapy of cancer has been limited by the considerable toxicity associated with such drugs. To address this challenge, we developed a smart theranostic prodrug system that combines a fluorophore and an anticancer drug, specifically 6-diazo-5-oxo-l-norleucine (DON), using a thioketal linkage (TK). This system enables imaging, chemotherapy, photodynamic therapy, and on-demand drug release upon radiation exposure. The optimized prodrug, DON-TK-BM3, incorporating cyanine dyes as the fluorophore, displayed potent reactive oxygen species release and efficient tumor cell killing. Unlike the parent drug DON, DON-TK-BM3 exhibited no toxicity toward normal cells. Moreover, DON-TK-BM3 demonstrated high tumor accumulation and reduced side effects, including gastrointestinal toxicity, in mice. This study provides a practical strategy for designing prodrugs of metabolic inhibitors with significant toxicity stemming from their lack of tissue selectivity.

Objective To investigate the efficacy and the collective brain responses of acupuncture for post-stroke limb numbness.Methods A total of 24 patients with post-stroke limb numbness and 23 matched healthy participants were recruited.Both groups received coordinated acupuncture treatment to regulate their physical and mental state,three times a week for 4 weeks.Changes in limb numbness assessed by the visual analog scale(VAS)were observed before and after acupuncture.Functional magnetic resonance imaging(fMRI)scans were conducted on the participants during the scanning process with acupuncture intervention.Blood oxygen level-dependent(BOLD)data were collected in three states:resting state,acupuncture task state,and needle retention state.Data processing was performed using software such as fMRIPrep and brainIAK.Intersubject correlation(ISC)analysis was conducted to compare the ISC values of different brain regions between the two groups under different conditions.Results Acupuncture treatment significantly reduced the VAS scores of limb numbness.No significant changes in ISC values in both groups during resting state.Under acupuncture conditions,the patient group exhibited significantly increased ISC values in the left precentral gyrus,left paracentral lobule,left postcentral gyrus,left insula,left anterior cingulate gyrus,right middle frontal gyruss and right anterior cingulate gyrus.Under the needle retention condition,a slight negative activation was observed in the bilateral precentral gyrus of the control group.In the patient group,the ISC values of the left superior frontal gyrus,left precentral gyrus,left paracentral lobule,left precuneus,left postcentral gyrus,left insula,left anterior cingulate gyrus,left thalamus,right superior frontal gyrus,right middle frontal gyrus,right precentral gyrus,and right anterior cingulate gyrus showed a significant increase compared to the resting state.Conclusion Acupuncture can improve post-stroke limb numbness symptoms and induce a specific pattern of collective brain activation in patients.This activation involves the precentral gyrus,postcentral gyrus,thalamus,insula,frontal lobe and cingulate gyrus.Acupuncture may enhance the processing of sensory information by activating the somatosensory-motor network and the cortical-thalamic-cortical loop.It may also modulate the central executive network and the descending pain control pathway to promote the relief of pain symptoms.

Background::Heterotaxy (HTX) is a thoracoabdominal organ anomaly syndrome and commonly accompanied by congenital heart disease (CHD). The aim of this study was to analyze rare copy number variations (CNVs) in a HTX/CHD cohort and to examine the potential mechanisms contributing to HTX/CHD.Methods::Chromosome microarray analysis was used to identify rare CNVs in a cohort of 120 unrelated HTX/CHD patients, and available samples from parents were used to confirm the inheritance pattern. Potential candidate genes in CNVs region were prioritized via the DECIPHER database, and PNPLA4 was identified as the leading candidate gene. To validate, we generated PNPLA4-overexpressing human induced pluripotent stem cell lines as well as pnpla4-overexpressing zebrafish model, followed by a series of transcriptomic, biochemical and cellular analyses. Results::Seventeen rare CNVs were identified in 15 of the 120 HTX/CHD patients (12.5%). Xp22.31 duplication was one of the inherited CNVs identified in this HTX/CHD cohort, and PNPLA4 in the Xp22.31 was a candidate gene associated with HTX/CHD. PNPLA4 is expressed in the lateral plate mesoderm, which is known to be critical for left/right embryonic patterning as well as cardiomyocyte differentiation, and in the neural crest cell lineage. Through a series of in vivo and in vitro analyses at the molecular and cellular levels, we revealed that the biological function of PNPLA4 is importantly involved in the primary cilia formation and function via its regulation of energy metabolism and mitochondria-mediated ATP production. Conclusions::Our findings demonstrated a significant association between CNVs and HTX/CHD. Our data strongly suggested that an increased genetic dose of PNPLA4 due to Xp22.31 duplication is a disease-causing risk factor for HTX/CHD.

Objective To investigate the relationship between exposure to famine in early life stage and hypertension phenotype and grade in middle and old age. Methods People born between 1951 and 1965 in the 2015 China Health and Elderly Care Follow-up Survey were included in the study, and were divided into unexposed group, fetal exposed group, childhood exposed group and adolescent exposed group according to the time of famine occurrence and birth year of the participants. Logistic regression model was used to explore the effects of different famine exposure periods in early life stage on hypertension classification （including normal high value, grade I, grade II and grade III） and phenotype （including isolated systolic hypertension[ISH], isolated diastolic hypertension [IDH] and combined systolic and diastolic hypertension [SDH]）. Results Compared with unexposed group, fetal famine exposure （OR=1.59, 95% CI :1.10-2.30）, childhood famine exposure （OR=1.67, 95% CI :1.04-2.70） and adolescent famine exposure （OR=3.42, 95% CI : 2.51-4.66） were the risk factors for ISH. Only famine exposure during adolescence （OR=1.54, 95% CI: 1.07-2.21） was a risk factor for SDH. In addition, fetal famine exposure （OR=1.41, 95% CI: 1.05-1.89） and adolescent famine exposure （OR=2.22 , 95% CI: 1.71-2.88） were risk factors for developing grade I hypertension. Famine exposure in childhood （OR=2.45, 95% CI: 1.21-4.94） and famine exposure in adolescence （OR=2.45, 95% CI: 1.44-4.19） were risk factors for grade 2 hypertension. Conclusion Famine exposure in early life stage was associated with the phenotype and grade of hypertension. Therefore, balanced nutrition in early life is important to prevent hypertension in adulthood.

Objective To express the sucrose isomerase(SI) fused with the tetrameric coiled-coil domain of the cell surface protein tetrabrachion(TdoT),and study the enzymatic properties of the recombinant enzymes.Methods The gene of SI fused with TdoT at the N/C terminus was cloned into the expression vectors respectively to construct the recombinant expression vectors pET-24a-TdoT-SI and pET-24b-SI-TdoT,which were transformed into E.coli BL21(DE3) and induced to express recombinant enzymes.The enzymatic properties and product specificity of the purified recombinant enzymes were studied.Results TdoT-SI and SI-TdoT were expressed as inclusion bodies with catalytic activity,while SI inclusion bodies without TdoT showed no catalytic activity.The results of enzymatic property analysis showed that the optimum reaction temperature for TdoT-SI and SI-TdoT active inclusion bodies was 40 ℃,and the optimum reaction pH was 5.5 and 5.0,respectively.The K_m of TdoT-SI active inclusion bodies was(103.9±9.5) mmol/L and the k_(cat)/K_m was(0.06±0.002) L/(mmol·s),while the K_m of SI-TdoT active inclusion bodies was(54.4±6.6) mmol/L and the k_(cat)/K_m was(0.03±0.002) L/(mmol·s).The results of product specificity analysis exhibited that the proportion of isomaltulose in the product did not change significantly,while the proportion of trehalose decreased,and the proportion of monosaccharides increased with increasing reaction temperature.Conclusion The active inclusion bodies of SI fused with coiled-coil domain were successfully prepared by fusion expression technology.As a novel self-immobilized enzyme,it has the advantage of simultaneous expression and immobilization,which provides a new strategy for large-scale preparation and efficient utilization of recombinant SI.

Pyroptosis is a type of programmed cell death distincted from apoptosis and necrosis, which is accompanied by the lysis of cell membranes and the release of cell contents. Pyroptosis occurs as mediated by Gasdermin protein family and is dependent on the activity of caspase. GSDME is one of the most important members of the Gasdermin protein superfamily. GSDME-mediated pyroptosis relies on the activity of caspase-3. In recent years, with further research on pyroptosis, the mechanism of GSDME-induced pyroptosis is becoming clear. Numerous studies have shown that GSDME-mediated pyroptosis plays an important role in the occurrence and development of tumors, as well as chemotherapy resistance. However, GSDME-mediated pyroptosis has no specificity and can induce pyroptosis of normal cells in the body while inducing tumor cell pyroptosis, thus causing different degrees of damage to various organs of the body. Further study on the mechanism of GSDME-induced pyroptosis, the role of GSDME in malignant tumors and the adverse reactions of chemotherapy can provide new ideas for tumor monitoring, treatment and prognosis judgment.

Objective:To investigate the status and influencing factors of psychological resilience of ophthalmic hospital Operating Room nurses, and to explore the correlation between psychological resilience and career development of Operating Room nurses.Methods:This is a cross-sectional study. From October to December 2021, 40 Operating Room nurses from Eye Hospital of Wenzhou Medical University were selected by convenience sampling method and investigated using general information questionnaire, Connor-Davidsion Resilience Scale (CD-RISC) and Career Development Questionnaire. Univariate analysis was used to explore the influencing factors of psychological resilience of Operating Room nurses. Pearson correlation analysis was employed to explore the correlation between psychological resilience and career development. A total of 40 questionnaires were distributed in this study, and 40 valid questionnaires were recovered, with an effective recovery rate of 100.0%.Results:The CD-RISC score of the Operating Room nurses was (90.40±9.27) , the score of Career Development Questionnaire was (112.5±9.65) . Univariate analysis results showed that age, professional title and working years were the influencing factors of psychological resilience of ophthalmic hospital Operating Room nurses ( P<0.01) . Pearson correlation analysis results showed that psychological resilience of Operating Room nurses was positively correlated with career development ( r=0.762, P<0.05) . Conclusions:The psychological resilience of ophthalmic hospital Operating Room nurses is at a medium level, and age, professional title and working years are its influencing factors. Operating Room nurses with high psychological resilience have better career development. Nursing managers should formulate personalized study and career plans, so as to promote nurses ' career development.

Enzalutamide (ENZ) is a second-generation androgen receptor (AR) antagonist used for the treatment of castration-resistant prostate cancer (CRPC) and reportedly prolongs survival time within a year of starting therapy. However, CRPC patients can develop ENZ resistance (ENZR), mainly driven by abnormal reactivation of AR signaling, involving increased expression of the full-length AR (ARfl) or dominantly active androgen receptor splice variant 7 (ARv7) and ARfl/ARv7 heterodimers. There is currently no efficient treatment for ENZR in CRPC. Herein, a small molecule LLU-206 was rationally designed based on the ENZ structure and exhibited potent inhibition of both ARfl and constitutively active ARv7 to inhibit PCa proliferation and suppress ENZR in CRPC. Mechanically, LLU-206 promoted ARfl/ARv7 protein degradation and decreased ARfl/ARv7 heterodimers through mouse double minute 2-mediated ubiquitination. Finally, LLU-206 exhibited favorable pharmacokinetic properties with poor permeability across the blood-brain barrier, leading to a lower prevalence of adverse effects, including seizure and neurotoxicity, than ENZ-based therapies. In a nutshell, our findings demonstrated that LLU-206 could effectively inhibit ARfl/ARv7-driven CRPC by dual-targeting of ARfl/ARv7 heterodimers and protein degradation, providing new insights for the design of new-generation AR inhibitors to overcome ARfl/ARv7-driven CRPC.

The data of a case of congenital subglottic stenosis (C-SGS) who underwent slide laryngotracheoplasty in the Center for Respiratory Intervention, Children′s Hospital Affiliated to Shandong University in December 2021 was analyzed retrospectively.The patient was a girl aged 2 months and 15 days.She visited the hospital 23 days after tracheotomy due to dyspnea for more than 2 months.The bronchoscopy and annular cartilage B ultrasound results suggested subglottic stenosis and no scar hyperplasia.Based on the medical history, the child was diagnosed with C-SGS.Slide laryngotracheoplasty was performed 2 weeks after admission, and the tracheotomy tube was removed after surgery.The child was followed up 2 months after surgery, and she recovered well with no dyspnea.The study results suggest that early and safe slide laryngotracheoplasty after definite diagnosis can provide immediate and sufficient airway space for C-SGS patients, and protect their voice and swallowing function.