ObjectiveTo investigate the mechanisms by which Huanglian Jiedutang （HLJDT） modulates microglial （MG） phenotypes through the sialic acid-binding Ig-like lectin 2 （SIGLEC2/CD22）/Src-homology-2-domain-containing protein tyrosine phosphatase-1 （SHP-1）/phosphorylated protein kinase B （p-Akt） signaling pathway， thereby promoting myelin repair and alleviating agitation-like behaviors in vascular dementia （VAD）. MethodsSixty C57BL/6J mice were randomly assigned to a sham （normal） group， model group， HLJDT low-， medium-， and high-dose groups （2.5， 5， and 10 g·kg^-1·d^-1）， and a risperidone group （2 mg·kg^-1·d^-1）， with 10 mice per group. VAD was induced by bilateral common carotid artery stenosis （BCAS）. From day 42， mice received drug interventions for 2 weeks. Agitation-like behaviors were assessed using the resident-intruder test. After behavioral testing， ventrolateral part of the ventromedial hypothalamus （VMHvl） tissues were collected. Western blot was used to measure protein levels of myelin oligodendrocyte glycoprotein （MOG）， myelin basic protein （MBP）， proteolipid protein （PLP）， inducible nitric oxide synthase （iNOS）， arginase-1 （Arg1）， CD86， CD206， and CD22， SHP-1， and p-Akt. Immunofluorescence was used to evaluate myelin-associated glycoprotein （MAG） intensity and the proportion of iNOS⁺/ionized calcium-binding adapter molecule 1 （Iba1）⁺ cells. ELISA was used to detect tumor necrosis factor-α （TNF-α）， interleukin （IL）-6， and IL-1β. ResultsCompared with the normal group， the model group exhibited markedly increased biting and aggressive behaviors and shortened attack latency （P<0.01）. MOG， MBP， and PLP protein levels and MAG fluorescence intensity were significantly reduced （P<0.05， P<0.01）. INOS and CD86 expression and TNF-α， IL-6， and IL-1β levels were significantly elevated （P<0.01）. CD22 and SHP-1 expression increased significantly （P<0.01）， whereas p-Akt expression decreased （P<0.01）. Compared with the model group， the medium- and high-dose HLJDT groups and the risperidone group showed markedly reduced biting and aggression （P<0.05， P<0.01） and prolonged attack latency （P<0.01）. MOG， MBP， and PLP levels and MAG fluorescence intensity were significantly increased （P<0.05， P<0.01）. INOS， CD86， TNF-α， IL-6， and IL-1β levels decreased significantly （P<0.05， P<0.01）. CD22 and SHP-1 expression decreased， while p-Akt expression increased significantly （P<0.05， P<0.01）. ConclusionHLJDT may modulate CD22/SHP-1/p-Akt signaling in the VMHvl， promote the shift of MG toward an anti-inflammatory and phagocytic phenotype， enhance myelin repair， and improve agitation-like behaviors in VAD mice.

BACKGROUND:Black phosphorus has a high degree of homology with human bone,so it has been extensively studied in the field of bone tissue engineering in recent years.Since 2014,two-dimensional black phosphorus materials have garned significant attention in the field of biomedicine due to their excellent exceptional physical,chemical,and biological properties. OBJECTIVE:To summarize the advancements made in black phosphorus-based nanomaterials for bone tissue engineering,focus on the synthesis methods,osteogenic characteristics,and applications in biomaterials pertaining to two-dimensional black phosphorus nanomaterials. METHODS:Chinese and English key words were"black phosphorus,bone tissue engineering,bone defect,bone regeneration,osteogenesis."Relevant articles in PubMed and CNKI databases from January 2014 to December 2023 were searched.After exclusion and screening,96 articles were analyzed. RESULTS AND CONCLUSION:Black phosphorus nanomaterials play an important role in bone tissue engineering due to their good biocompatibility,biodegradability,photothermal action,antibacterial ability,drug loading performance,and special osteogenic effect,and are ideal candidate materials for promoting bone regeneration.The preparation of black phosphorus nanomaterials is mainly a top-down top-layer stripping method.The main principle is to weaken the van der Waals force between the black phosphorus layers by physical or chemical means to obtain a single or less layer of phosphanse,that is,black phosphorus nanosheets or quantum dots.Black phosphate-based nanocomposites are mainly divided into hydrogels,3D printing scaffolds,composite scaffolds,electrospinning,bionic periosteum,microspheres,and bionic coatings.The research of nano-black phosphorus in bone tissue engineering is in its infancy,and still faces many challenges:the behavior of black phosphorus in vivo and the interaction mechanism with various biomolecules need to be further studied.The long-term potential toxicity of black phosphorus is unknown.The manufacturing process for black phosphorus is difficult to control.Therefore,how to develop uniform size,safe,reliable,and efficient nano black phosphorus and transform it into clinical application requires interdisciplinary research on modern biomedical technology,physicochemical technology,and precision manufacturing technology.

ObjectiveTo observe and compare the effects of different concentrations of cyclophosphamide (CTX) in inducing premature ovarian insufficiency (POI) model in mice and investigate the mechanism of injury. MethodsThirty-two 6~8-week-old female C57BL/6J mice were randomly divided into four groups (n=8 per group) using a weight-based block randomization method. The POI model was established via a single intraperitoneal injection of 75 mg/kg cyclophosphamide (CTX), 120 mg/kg CTX, 120 mg/kg CTX + 12 mg/kg Busulfan, or an equivalent volume of normal saline (control). Ovarian coefficients, serum estradiol (E₂) and follicle-stimulating hormone (FSH) levels were measured. Western blotting was performed to assess changes in ovarian expression levels of NAD-dependent deacetylase sirtuin-5 (SIRT5) and forkhead box O3a (FOXO3a) under different modeling conditions. After determining the optimal CTX concentration for modeling, an additional forty 6~8-week-old femal C57BL/6J mice were randomly divided into five groups (n=8 per group) using a weight-based block randomization method: saline control, 120 mg/kg CTX sampling at 1, 2, 7, or 14 days after modeling. Western blotting was used to evaluate temporal changes of ovarian SIRT5 and FOXO3a protein expression. ResultsCompared with the saline control, all concentrations of CTX (75 mg/kg CTX, 120 mg/kg CTX) and 120 mg/kg CTX + 12 mg/kg Busulfan induced POI injury in mice. The 120 mg/kg CTX group exhibited smaller changes in ovarian coefficients (P<0.001) and E₂ levels (P<0.05), whereas the 120 mg/kg CTX + 12 mg/kg Busulfan group showed rough and reduced luster fur, sluggish response and was in the worst state. Compared with the saline control group, FOXO3a expression was significantly down-regulated (P<0.05), while SIRT5 remained unchanged in the 75 mg/kg CTX group (P>0.05). In contrast, both SIRT5 (P<0.05) and FOXO3a (P<0.05) were significantly down-regulated in the 120 mg/kg CTX group. Further analysis revealed that on day 2 and 7 after 120 mg/kg CTX modeling, the expressions of SIRT5 (P<0.01) and FOXO3a (P<0.001) were significantly down-regulated, with the largest decrease observed on day 7 (SIRT5, P<0.000 1; FOXO3a, P<0.000 1). ConclusionOvarian injury in the POI model induced by 120 mg/kg CTX is milder than that in the POI model induced by 75 mg/kg CTX. Moreover, the expression changes of SIRT5 and FOXO3a are most significant on day 7 after modeling induced by 120 mg/kg CTX, which may be related to the inhibition of the SIRT5-FOXO3a signaling pathway.

ObjectiveTo investigate the ameliorative effects of quercetin on neuropsychiatric symptoms associated with vascular dementia （VaD） and to elucidate the molecular mechanism， specifically whether quercetin inhibits pro-inflammatory activation of microglia by modulation of the receptor-interacting serine/threonine-protein kinase 1 （RIPK1）/NOD-like receptor protein 3 （NLRP3）/Caspase-1 signaling pathway， thereby promoting myelin repair in the medial prefrontal cortex （mPFC）. MethodsA C57BL/6J mouse model of VaD with neuropsychiatric symptoms was established by bilateral common carotid artery stenosis （BCAS） combined with chronic restraint stress （CRS）. Mice were randomly divided into a sham group， a model group， low-dose， medium-dose， and high-dose quercetin groups （30， 60， 120 mg·kg^-1·d^-1）， and a fluoxetine group （10 mg·kg^-1·d^-1）. After intervention， depressive- and anxiety-like behaviors were assessed by the sucrose preference test （SPT）， forced swim test （FST）， open field test （OFT）， and elevated plus maze （EPM）. mPFC tissue was collected. Immunofluorescence （IF） was used to detect myelin basic protein （MBP） expression and microglial morphology. Western blot was used to measure the protein level of MBP， myelin oligodendrocyte glycoprotein （MOG）， myelin-associated glycoprotein （MAG）， inducible nitric oxide synthase （iNOS）， CD86， RIPK1， phosphorylated RIPK1 （Ser166）， NLRP3， and Caspase-1. Enzyme-linked immunosorbent assay （ELISA） was used to determine the level of tumor necrosis factor-alpha （TNF-α）， interleukin-6 （IL-6）， and interleukin-1β （IL-1β）. ResultsCompared with the sham group， the model group exhibited significant depressive- and anxiety-like behaviors （P<0.01）， significantly decreased protein expression of MBP， MOG， and MAG in the mPFC （P<0.01）， activated microglia （characterized by enlarged cell bodies， reduced protrusions， and upregulated iNOS and CD86 expressions， P<0.01）， and significantly elevated p-RIPK1/RIPK1 ratio， NLRP3， Caspase-1 protein expression， and level of TNF-α， IL-6， and IL-1β （P<0.01， P<0.05）. Compared with the model group， the quercetin treatment （especially at medium and high doses） significantly ameliorated these behavioral abnormalities （P<0.05， P<0.01）， increased the expression of MBP （protein and fluorescence intensity）， MOG， and MAG in the mPFC （P<0.05， P<0.01）， suppressed excessive microglial activation （characterized decreased cell bodies， increased protrusions， and downregulated iNOS and CD86 expressions， P<0.01）， and significantly reduced the p-RIPK1/RIPK1 ratio， NLRP3， Caspase-1 protein expression， and inflammatory cytokine levels （P<0.01）. ConclusionQuercetin effectively alleviates neuropsychiatric symptoms in VaD mice. Its mechanism may be associated with the inhibition of microglial inflammatory activation mediated by the RIPK1/NLRP3/Caspase-1 signaling pathway， thereby promoting myelin repair in the mPFC region.

Ovarian cancer is the most lethal malignancy affecting the female reproductive system. Pharmacological inhibitors targeting CDK4/6 have demonstrated promising efficacy across various cancer types. However, their clinical benefits in ovarian cancer patients fall short of expectations, with only a subset of patients experiencing these advantageous effects. This study aims to provide further clinical and biological evidence for antineoplastic effects of a CDK4/6 inhibitor (TQB4616) in ovarian cancer and explore underlying mechanisms involved. Patient-derived ovarian cancer organoid models were established to evaluate the effectiveness of TQB3616. Potential key genes related to TQB3616 sensitivity were identified through RNA-seq analysis, and TRIM4 was selected as a candidate gene for further investigation. Subsequently, co-immunoprecipitation and GST pull-down assays confirmed that TRIM4 binds to hnRNPDL and promotes its ubiquitination through RING and B-box domains. RIP assay demonstrated that hnRNPDL binded to CDKN2C isoform 2 and suppressed its expression by alternative splicing. Finally, in vivo studies confirmed that the addition of siTRIM4 significantly improved the effectiveness of TQB3616. Overall, our findings suggest that TRIM4 modulates ubiquitin-mediated degradation of hnRNPDL and weakens sensitivity to CDK4/6 inhibitors in ovarian cancer treatment. TRIM4 may serve as a valuable biomarker for predicting sensitivity to CDK4/6 inhibitors in ovarian cancer.
Humans ; Female ; Ovarian Neoplasms/pathology* ; Animals ; Tripartite Motif Proteins/genetics* ; Mice ; Cyclin-Dependent Kinase 4/antagonists & inhibitors* ; Cell Line, Tumor ; Cyclin-Dependent Kinase 6/antagonists & inhibitors* ; Protein Kinase Inhibitors/pharmacology* ; Ubiquitin/metabolism* ; Xenograft Model Antitumor Assays ; Ubiquitination ; Antineoplastic Agents/pharmacology*

Objective To investigate the expression levels of micro RNA-651(miR-651)and the transcrip-tion factor Forkhead Box N2(FOXN2)in lung cancer tissues and to analyse their relationship with patients' prognosis after lobectomy.Methods A total of 98 patients with lung cancer who underwent lobectomy from January 2020 to December 2021 in Nantong Cancer Hospital were selected as research objects,the lung cancer tissue specimens and corresponding paracancerous tissue specimens of patients were collected,the expression levels of miR-651 and FOXN2 were detected by real-time fluorescence quantitative PCR(qPCR),and their re-lationship with the prognosis of patients after lobectomy was analyzed.Results At 2-year follow-up,19(19.39％)of 98 patients had recurrent metastasis,and the median time to recurrent metastasis was 20 months(95％CI:18.626--21.374).qPCR assay showed that the expression levels of miR-651 and FOXN2 in lung cancer tissues were significantly lower than those in paracancerous tissues(P＜0.05).miR-651 and FOXN2 expression levels were significantly correlated with clinicopathological features such as tumour size,TNM stage and lymph node metastasis(P＜0.05).The results of univariate Cox regression analysis showed that the percentages of maximum tumor diameter ≥3 cm,TNM stage Ⅱ,lymph node metastasis,miR-651 low ex-pression and FOXN2 low expression were higher in the poor prognosis group than those in the good prognosis group(P＜0.05).Multifactorial Cox regression analysis showed that maximum tumor diameter ≥ 3 cm(HR=3.185,95％CI:1.105-9.182),TNM stage Ⅱ(HR=5.167,95％CI:1.939-13.771),lymph node metastasis(HR=2.860,95％CI:1.115-7.338),low miR-651 expression(HR=5.345,95％CI:1.845-15.483)and low FOXN2 expression(HR=5.404,95％CI:1.744-16.745)were independent risk factors for the prognosis of lung cancer patients after lobectomy(P＜0.05).Conclusion The expression levels of miR-651 and FOXN2 were reduced in the tissues of lung cancer patients,and both were associated with tumour size,TNM stage,lymph node metastasis,and patients' prognosis after lobectomy.

ObjectiveThis study aims to investigate the effect of Huanglian Jiedutang on brain iron metabolism disorders， myelin damage， and aggressive behavior in vascular dementia （VaD） mice by regulating the F-box leucine-rich repeat protein 5 （FBXL5）/iron regulatory protein 2 （IRP2） pathway. MethodSixty C57BL/6J mice were randomly divided into six groups： sham operation group， model group， risperidone group （2 mg·kg^-1·d^-1）， and low-dose， medium-dose， and high-dose groups of Huanglian Jiedutang （0.25， 0.5， 1 g·kg^-1·d^-1）， with 10 mice in each group. The VaD model was established by bilateral carotid artery stenosis （BCAS）. Drug intervention was administered for two weeks starting from the seventh week. Behavioral assessments， including the touch escape and resident-intruder tests， were conducted in the ninth week. After the behavioral tests， ventromedial hypothalamus ventrolateral （VMHvl） tissue samples were collected. Western blot was used to detect the expression of myelin-associated glycoprotein （MAG）， myelin oligodendrocyte glycoprotein （MOG）， myelin basic protein （MBP）， 4-hydroxynonenal （4-HNE）， glutathione peroxidase 4 （GPX4）， transferrin receptor 1 （TFR1）， ferritin light chain （Ft-L）， ferroportin 1 （FPN1）， FBXL5， and IRP2. Immunofluorescence was used to measure MBP fluorescence intensity. Transmission electron microscopy was employed to observe ultrastructural changes in the myelin sheath. Perl's staining was used to detect tissue iron deposition. Malondialdehyde （MDA） and superoxide dismutase （SOD） levels were measured by enzyme linked immunosorbent assay （ELISA）. ResultCompared to the sham operation group， the model group exhibited a significant increase in biting， aggression， and irritability scores， along with a reduced latency of the attack （P<0.01）. The expression levels of MAG， MOG， and MBP， as well as the fluorescent density of MBP， were significantly decreased （P<0.01）. Disordered myelin ultrastructure， increased Ft-L and TFR1 expression， decreased FPN1 expression， and elevated iron deposition were observed （P<0.01）. Antioxidants GPX4 and SOD were reduced， while 4-HNE and lipid peroxidation product MDA were increased （P<0.01）. FBXL5 protein expression decreased， and IRP2 protein expression increased （P<0.01）. Compared with the model group， in the middle-dose and high-dose groups of Huanglian Jiedutang and the risperidone group， the number of bites， aggressive behaviors， and irritability scores were reduced （P<0.05， P<0.01）， while the latency of the attack increased （P<0.01）. In the middle-dose and high-dose groups of Huanglian Jiedutang， the expression levels of MAG， MOG， and MBP， as well as MBP immunofluorescence increased （P<0.01）. The ultrastructure was orderly arranged. Ft-L and TFR1 expression decreased； FPN1 expression increased， and iron deposition was reduced （P<0.01）. GPX4 and SOD expression levels increased， and 4-HNE and MDA expression levels decreased （P<0.01）. FBXL5 protein expression increased， and IRP2 protein expression decreased （P<0.01）. ConclusionHuanglian Jiedutang may alleviate brain iron metabolism disorders， myelin damage， and aggressive behavior in VaD mice by regulating FBXL5/IRP2 expression in the VMHvl region.

OBJECTIVE To calculate the cost of centralized dispensing of four categories of drugs （ordinary drugs， antibacterial drugs， hazardous drugs， and parenteral nutrition solutions） in pharmacy intravenous admixture service （PIVAS）， and provide reference for setting charging standards for relevant departments. METHODS The operating costs of PIVAS in 12 medical institutions from Shaanxi province were collected through questionnaire survey， including labor costs， medical and health material costs， fixed asset depreciation and repair costs， water and electricity costs， and management costs. The operation time allocation coefficient method and workload allocation coefficient method were comprehensively used to allocate the above costs， and the unit preparation costs of four categories of drugs were calculated. RESULTS The average annual total costs of dispensing ordinary drugs， antibacterial drugs， hazardous drugs， and parenteral nutrition solutions in Shaanxi province were （2 195 900.25±1 680 893.73） yuan， （746 341.59±725 839.39） yuan， （331 420.15±183 258.83） yuan， and （330 322.68±277 281.70） yuan， respectively， with labor costs accounting for the highest proportion， averaging 85.49%. The costs of dispensing a set of ordinary drugs， antibacterial drugs， and hazardous drugs were 5.89， 7.60， and 14.37 yuan， respectively； the cost of dispensing one bag of parenteral nutrition solution was 32.15 yuan （excluding the cost of disposable intravenous nutrition bags）. CONCLUSIONS The cost calculation method and data of different types of intravenous drugs obtained in this study can provide reference for relevant departments to formulate and adjust PIVAS fee standards.

OBJECTIVE To calculate the cost of centralized dispensing of four categories of drugs （ordinary drugs， antibacterial drugs， hazardous drugs， and parenteral nutrition solutions） in pharmacy intravenous admixture service （PIVAS）， and provide reference for setting charging standards for relevant departments. METHODS The operating costs of PIVAS in 12 medical institutions from Shaanxi province were collected through questionnaire survey， including labor costs， medical and health material costs， fixed asset depreciation and repair costs， water and electricity costs， and management costs. The operation time allocation coefficient method and workload allocation coefficient method were comprehensively used to allocate the above costs， and the unit preparation costs of four categories of drugs were calculated. RESULTS The average annual total costs of dispensing ordinary drugs， antibacterial drugs， hazardous drugs， and parenteral nutrition solutions in Shaanxi province were （2 195 900.25±1 680 893.73） yuan， （746 341.59±725 839.39） yuan， （331 420.15±183 258.83） yuan， and （330 322.68±277 281.70） yuan， respectively， with labor costs accounting for the highest proportion， averaging 85.49%. The costs of dispensing a set of ordinary drugs， antibacterial drugs， and hazardous drugs were 5.89， 7.60， and 14.37 yuan， respectively； the cost of dispensing one bag of parenteral nutrition solution was 32.15 yuan （excluding the cost of disposable intravenous nutrition bags）. CONCLUSIONS The cost calculation method and data of different types of intravenous drugs obtained in this study can provide reference for relevant departments to formulate and adjust PIVAS fee standards.

Objective:To investigate the efficacy and safety of daratumumab (Dara) - combination regimens for newly diagnosed multiple myeloma (NDMM).Methods:A retrospective case series study was conducted. The clinical data of 34 patients with NDMM receiving treatment regimen including Dara from Qilu Hospital of Shandong University, Yantai Yuhuangding Hospital, Huangdao Branch of Affiliated Hospital of Qingdao University and Taian City Central Hospital between April 2020 and March 2022 were retrospectively collected. The efficacy, survival and adverse reactions of patients were analyzed. Cox proportional risk model was used to analyze the factors affecting overall survival (OS) and minimal residual disease (MRD) turning negative.Results:Among 34 patients with NDMM, there were 19 males and 15 females, with 21 cases aged < 65 years and 13 cases aged ≥65 years. The median follow-up duration [ M ( Q1, Q3)] was 22 months (19 months, 26 months), the median of Dara treatment cycles was 7 (5, 11), and the overall response rate (ORR) reached 97.1% (33/34). There were statistically significant differences in the optimal efficacy of patients stratified by receiving hematopoietic stem cell transplantation or not and receiving different treatment cycles (all P ≤ 0.05), while there were no statistically significant differences in patients stratified by other clinical features (all P > 0.05). The 1-year progression-free survival rate was 79.4% and the 1-year OS rate was 94.1%. Multivariate Cox regression analysis showed that the cycle number of treatment regimens containing Dara was an independent influencing factor of MRD turning negative (6 cycles vs. 2 cycles, HR = 0.267, 95% CI: 0.076-0.935, P = 0.039); age ≥ 65 years was an independent risk factor for OS ( HR = 35.313, 95% CI: 1.709-729.669, P = 0.021). The incidence of hematological adverse reactions grade 3 or above was 20.6% (7/34), and the non-hematological adverse reactions primarily included infection [44.1% (15/34)] and edema of extremity and trunk [41.2% (14/34)]. Conclusions:The Dara-based regimens for NDMM exhibit a high ORR. The remission depth accelerated with the increasing number of treatment cycle, and the adverse reactions are mild.