ObjectiveThe malaria parasites remodel the host erythrocyte structure by exporting parasite proteins that interact with the membrane skeleton proteins of red blood cells (RBCs), facilitating their intracellular survival and pathogenicity. Skeleton-binding protein 1 (SBP1) is a conserved exported protein across Plasmodium species. In Plasmodium falciparum, SBP1 has been reported to interact with erythrocyte membrane skeleton proteins 4.1R and spectrin, while its contribution to erythrocyte remodeling and parasite virulence in Plasmodium berghei (Pb) remains unclear. This study aims to determine whether PbSBP1 associates with the host cytoskeletal protein 4.1R and to investigate its role in the remodeling of host RBCs and the pathogenicity of Plasmodium berghei. MethodsIn Plasmodium berghei, the relationship between PbSBP1 and the erythrocyte cytoskeletal protein 4.1R was examined using co-immunoprecipitation. A Pbsbp1 gene knockout mutant of Plasmodium berghei (Pbsbp1∆) was generated based on the principle of double crossover homologous recombination. The deformability of erythrocytes infected with Pbsbp1∆ parasites was assessed using microfluidic methods. Microchannels with an array of cylindrical pillars were used to detect modifications in infected RBC deformability. The infected RBCs were squashed between the rows and recovered between the columns and the transit velocity (μm/s) of infected RBCs travelling through the microchannel was recorded. The component of the erythrocyte membrane skeleton junctional complex, tropomodulin (TMOD), was fluorescently labeled, and the cytoskeletal network of infected erythrocytes was imaged using super-resolution stochastic optical reconstruction microscopy (STORM) to analyze ultrastructural changes in the cytoskeleton of wild-type (WT) and Pbsbp1∆-infected erythrocytes. Actin-based junctional complexes were displayed as individual clusters by the labeled TMOD in the STORM images, and the cluster densities and distances between adjacent clusters of infected RBCs were calculated. Additionally, rodent malaria models (BALB/c mice) and experimental cerebral malaria models (C57BL/6 mice) were employed to monitor the growth of Pbsbp1∆ and WT parasites during the intraerythrocytic stage and their capacity to induce cerebral malaria in mice. ResultsPbSBP1 may participate in the remodeling of infected erythrocytes through direct or indirect interaction with the erythrocyte cytoskeletal protein 4.1R. Microfluidic assays revealed that the deformability of erythrocytes infected with Pbsbp1∆ parasites was significantly enhanced compared to those infected with WT parasites. STORM imaging further demonstrated that the ultrastructure of the erythrocyte cytoskeleton in Pbsbp1∆-infected cells was altered relative to that in WT-infected erythrocytes. The distances between nearest neighbors of clusters had a tendency to increase while the cluster densities were decreased in Pbsbp1∆-infected RBCs compared to WT-infected RBCs. Subsequent phenotypic analysis indicated that the growth rate of Pbsbp1∆ parasites during the intraerythrocytic stage was significantly slower than that of WT parasites, and their ability to induce cerebral malaria in mice was also attenuated. These findings suggest that PbSBP1 is involved in the remodeling of the erythrocyte membrane skeleton, likely through its direct or indirect interaction with protein 4.1R, thereby regulating the deformability of infected erythrocytes and influencing the pathogenicity of the blood-stage parasites. ConclusionThis study establishes a role for PbSBP1 in host erythrocyte remodeling and parasite virulence, providing new research strategies for the prevention and treatment of malaria.

Objective This study explores the clinical correlation between different low-density lipoprotein cholesterol(LDL-C)levels and prognosis,providing evidence-based guidance for the development of personalized lipid-lowering goals.Methods Patients who underwent elective percutaneous coronary intervention(PCI)treatment at Beijing Anzhen Hospital from January 2020 to June 2023 and received lipid-lowering therapy with the addition of Evolocumab were selected.Based on the results of blood lipid rechecks 3 to 6 months after surgery,the patients were divided into five groups:low-density lipoprotein＜0.5 mmol/L,0.5 to＜1.0 mmol/L,1.0 to＜1.4 mmol/L,1.4 to＜1.8 mmol/L,and above 1.8 mmol/L.All patients were followed up for more than one year,and clinical conditions and major adverse cardiovascular events(MACE)were recorded.Results A total of 1 106 patients undergoing PCI were enrolled;after propensity score matching and exclusion of patients lost to follow up,550 remained(110 per group).During 12 months of follow-up,58 patients(10.5％)experienced a MACE,with incidence rising step-wise across LDL-C categories.In multivariable Cox models adjusted for age,sex,diabetes,hypertension,baseline LDL-C,follow-up LDL-C,estimated glomerular filtration rate(eGFR),and left ventricular ejection fraction,the hazard ratios[HR(95％CI)]for MACE,relative to the＜0.5 mmol/L group,were 1.810(0.507-6.454,P=0.361),3.036(0.945-9.749,P=0.062),5.228(1.737-15.735,P=0.003),7.708(2.633-22.565,P＜0.001)for LDL-C levels of 0.5 to＜1.0,1.0 to＜1.4,1.4 to＜1.8 and≥ 1.8 mmol/L,respectively.A restricted cubic spline model demonstrated a significant non-linear positive association between LDL-C and MACE(P-overall≤0.001;P-non-linear=0.008).Stratified analyses by age,sex,hypertension and diabetes showed consistent HR with no significant interactions(all P＞0.05).There were no statistically significant differences among the groups in the incidence of bleeding events,elevated creatinine levels,or abnormal liver function(all P＞0.05).Conclusions In patients using PCSK9 after PCI,there is a significant positive correlation between LDL-C levels and the risk of MACE,and no correlation was observed between different LDL-C levels and the risk of adverse events such as bleeding.

Objective·To investigate the role of interleukin-1β(IL-1β)in predicting the severity of oral and maxillofacial space infection(OMSI),and to explore the key mechanisms regulating IL-1β release,the critical immune cell subpopulations involved,and the intercellular communication networks among immune cells in OMSI patients.Methods·A total of 62 OMSI patients admitted to the Department of Oral Surgery,Shanghai Ninth People's Hospital,Shanghai Jiao Tong University School of Medicine,from January to November 2023 were enrolled,including 20 patients with moderate infection,21 with severe infection,and 21 with extremely severe infection.Logistic regression analysis was performed to identify risk factors for extremely severe infection,and receiver operating characteristic(ROC)curves were constructed to evaluate the ability of the above indicators to predict extremely severe infection.Peripheral blood mononuclear cells(PBMCs)from 2 patients in each group(moderate,severe and extremely severe)and 2 healthy controls(GSE224198)were analyzed using single-cell RNA sequencing(scRNA-seq)to identify key pro-inflammatory cell subtypes and genes,and to examine their changing trends with increasing infection severity.Cell-cell communication was assessed using CellChat.Quantitative real-time polymerase chain reaction(qPCR)and Western blotting were used to validate inflammasome activation levels in PBMCs.Results·Compared with patients with moderate and severe infections,levels of procalcitonin(PCT)(P<0.05)and IL-1β(P<0.05)were significantly elevated in patients with extremely severe infection.Logistic regression identified IL-1β as an independent risk factor for extremely severe infection(OR=1.814,95%CI 1.256?2.621,P=0.002).The area under the ROC curve(AUC)for the combined prediction of extremely severe infection using IL-1β and PCT was 0.943.scRNA-seq revealed continuous upregulation of NLRP3(NOD-like receptor family pyrin domain-containing 3)and IL1B gene expression in monocytes as infection severity increased,with intermediate monocytes being the main IL1B-expressing cell subtype.IL-1Β-IL-1R signaling,C-C motif chemokine ligand(CCL)and intercellular adhesion molecule(ICAM)signaling were significantly enhanced in monocytes.Macrophage migration inhibitory factor(MIF)signaling between T cells and monocytes also increased notably.With infection progression,the mRNA levels of NLRP3 and IL1B in peripheral blood rose steadily,and the protein levels of NLRP3,caspase-1 p20,apoptosis-associated speck-like protein containing a CARD(ASC)and IL-1β were persistently elevated.Conclusion·The combined levels of IL-1β and PCT at admission can effectively predict extremely severe OMSI.NLRP3 inflammasome activation is observed in PBMCs of OMSI patients.The elevation of IL-1β is closely associated with intermediate monocytes.Monocyte-mediated IL-1Β-IL-1R,CCL and ICAM signaling pathways,along with T cell-mediated MIF signaling pathways,collectively promote the inflammatory response.

Objective·To elevate the therapeutic effect of artesunate(ART)on NOD/Ltj mice(non-obese diabetic mice,the spontaneous models of Sj?gren syndrome),and explore its potential impact on the distribution of B lymphocyte subsets.Methods·ICR mice were used as the blank control group,and NOD/Ltj mice were randomly divided into disease and ART groups.NOD/Ltj mice and ICR mice were treated with ART(10 mg/kg)or its vehicle(0.5%CMC)by oral gavage every other day for 4 weeks.Body weight,salivary flow rate,submandibular gland index,and spleen index were measured.Cytokines in plasma,including interleukin-6(IL-6),interferon-γ(IFN-γ),and B-cell activating factor(BAFF)in serum,were detected by cytometric bead array(CBA).Hematoxylin-Eosin(H-E)staining of submandibular glands was used to observe the infiltration of lymphocyte.Flow cytometry was applied to analyze the distribution of B lymphocyte subsets in the spleen.The mRNA expression of Prdm1,Il-6r,Il-6,and Stat3 in spleen B lymphocytes was detected by RT-qPCR.The effect of ART on B cells was further detected by CCK-8 and Annexin V-FITC/PI staining by flow cytometry.Results·Compared to the disease group,ART significantly improved the symptoms of Sj?gren syndrome in NOD/Ltj mice.ART treatment also resulted in a reduction in the levels of BAFF,IL-6,and IFN-γ in the plasma(all P<0.05).Moreover,lymphocyte infiltration around the glandular ducts in the submandibular glands was greatly improved in the ART group compared with the disease group.Flow cytometry analysis revealed that the proportion of Na?ve B cells in the ART group was significantly increased compared with the disease group,along with a significant reduction in the proportions of double-negative B cells,switched memory B cells,and plasmablasts(all P<0.05).The relative mRNA expression levels of Prdm1,Il-6r,and Stat3 in the ART group were significantly lower than those in the disease group(all P<0.05).The CCK8 assay results showed that after 6 h of treatment,with the extension of the culture time,cell proliferation in the ART group was significantly inhibited;after 24 h of treatment,the number of apoptotic cells in the ART group was significantly higher than that in the control group(P<0.001).Conclusion·ART demonstrates therapeutic effects in NOD/Ltj mice,potentially through modulating the distribution of peripheral B lymphocyte subsets.It can inhibit the expression of Prdm1,thereby regulating the differentiation of B lymphocytes into plasma cells and plasmablasts.

Objective This study explores the clinical correlation between different low-density lipoprotein cholesterol(LDL-C)levels and prognosis,providing evidence-based guidance for the development of personalized lipid-lowering goals.Methods Patients who underwent elective percutaneous coronary intervention(PCI)treatment at Beijing Anzhen Hospital from January 2020 to June 2023 and received lipid-lowering therapy with the addition of Evolocumab were selected.Based on the results of blood lipid rechecks 3 to 6 months after surgery,the patients were divided into five groups:low-density lipoprotein＜0.5 mmol/L,0.5 to＜1.0 mmol/L,1.0 to＜1.4 mmol/L,1.4 to＜1.8 mmol/L,and above 1.8 mmol/L.All patients were followed up for more than one year,and clinical conditions and major adverse cardiovascular events(MACE)were recorded.Results A total of 1 106 patients undergoing PCI were enrolled;after propensity score matching and exclusion of patients lost to follow up,550 remained(110 per group).During 12 months of follow-up,58 patients(10.5％)experienced a MACE,with incidence rising step-wise across LDL-C categories.In multivariable Cox models adjusted for age,sex,diabetes,hypertension,baseline LDL-C,follow-up LDL-C,estimated glomerular filtration rate(eGFR),and left ventricular ejection fraction,the hazard ratios[HR(95％CI)]for MACE,relative to the＜0.5 mmol/L group,were 1.810(0.507-6.454,P=0.361),3.036(0.945-9.749,P=0.062),5.228(1.737-15.735,P=0.003),7.708(2.633-22.565,P＜0.001)for LDL-C levels of 0.5 to＜1.0,1.0 to＜1.4,1.4 to＜1.8 and≥ 1.8 mmol/L,respectively.A restricted cubic spline model demonstrated a significant non-linear positive association between LDL-C and MACE(P-overall≤0.001;P-non-linear=0.008).Stratified analyses by age,sex,hypertension and diabetes showed consistent HR with no significant interactions(all P＞0.05).There were no statistically significant differences among the groups in the incidence of bleeding events,elevated creatinine levels,or abnormal liver function(all P＞0.05).Conclusions In patients using PCSK9 after PCI,there is a significant positive correlation between LDL-C levels and the risk of MACE,and no correlation was observed between different LDL-C levels and the risk of adverse events such as bleeding.

Objective:To investigate the neuroprotective effect of non-invasive vagus nerve stimulation(nVNS)on traumatic brain injury(TBI).Methods:Male SD rats were used to prepare controlled cortical impact(CCI)rat mod-el.nVNS treatment was performed.The motor function of rats was detected by beam walking test.The water content of rats was evaluated by the dry-wet ratio of rat brain tissue.The content of lactate dehydrogenase(LDH)in cerebrospinal fluid was detected by commercial kit.The expression of bain-derived neurotrophic factor(BDNF)and nerve growth fac-tor(NGF)mRNA in rat cortex was detected by RT-qPCR.Results:After nVNS stimulation,CCI rats significantly im-proved the neurological function deficit,improved the motor ability,decreased the cerebral water content,decreased the level of LDH in cerebrospinal fluid,and upregulated the expression of BDNF and NGF mRNA in cortex.Conclusion:nVNS has a neuroprotective effect on TBI rats,and its mechanism may be related to the increased expression of BDNF and NGF.

Guided by the concept of quality by design(QbD), this study optimizes the calcination and quenching process of calcined Magnetitum and establishes the XRD characteristic spectra of calcined Magnetitum, providing a scientific basis for the formulation of quality standards. Based on the processing methods and quality requirements of Magnetitum in the Chinese Pharmacopoeia, the critical process parameters(CPPs) identified were calcination temperature, calcination time, particle size, laying thickness, and the number of vinegar quenching cycles. The critical quality attributes(CQAs) included Fe mass fraction, Fe~(2+) dissolution, and surface color. The weight coefficients were determined by combining Analytic Hierarchy Process(AHP) and the criteria importance though intercrieria correlation(CRITIC) method, and the calcination process was optimized using orthogonal experimentation. Surface color was selected as a CQA, and based on the principle of color value, the surface color of calcined Magnetitum was objectively quantified. The vinegar quenching process was then optimized to determine the best processing conditions. X-ray diffraction(XRD) was used to establish the characteristic spectra of calcined Magnetitum, and methods such as similarity evaluation, cluster analysis, and orthogonal partial least squares-discriminant analysis(OPLS-DA) were used to evaluate the quality of the spectra. The optimized calcined Magnetitum preparation process was found to be calcination at 750 ℃ for 1 h, with a laying thickness of 4 cm, a particle size of 0.4-0.8 cm, and one vinegar quenching cycle(Magnetitum-vinegar ratio 10∶3), which was stable and feasible. The XRD characteristic spectra analysis method, featuring 9 common peaks as fingerprint information, was established. The average correlation coefficient ranged from 0.839 5-0.988 1, and the average angle cosine ranged from 0.914 4 to 0.995 6, indicating good similarity. Cluster analysis results showed that Magnetitum and calcined Magnetitum could be grouped together, with similar compositions. OPLS-DA discriminant analysis identified three key characteristic peaks, with Fe_2O_3 being the distinguishing component between the two. The final optimized processing method is stable and feasible, and the XRD characteristic spectra of calcined Magnetitum was initially established, providing a reference for subsequent quality control and the formulation of quality standards for calcined Magnetitum.
X-Ray Diffraction/methods* ; Drugs, Chinese Herbal/chemistry* ; Quality Control ; Particle Size

As a new type of production factor that can empower the development of new quality productivity, the data element is an important engine to promote the high quality development of the industry. Traditional Chinese medicine(TCM) dispensing is the most basic work of TCM clinical pharmacy, and its quality directly affects the clinical efficacy of TCM. The integration of data elements and TCM dispensing can stimulate the innovation and vitality of the TCM dispensing industry and promote the high-quality and sustainable development of the industry. A large-scale, detailed, and systematic study on TCM dispensing was conducted. The innovative practice path of data fusion construction in the whole process of TCM dispensing was investigated by integrating the digital resources "nine full activities" of TCM dispensing, creating the digital dictionary of "TCM clinical information data elements", and exploring innovative applications of TCM dispensing driven by data and technology, so as to promote the standardized, digital, and intelligent development of TCM dispensing in medical health services. The research content of this project was successfully selected as the second batch of "Data element×" typical cases of National Data Administration in 2024, which is the only selected case in the field of TCM.
Medicine, Chinese Traditional/methods* ; Drugs, Chinese Herbal ; Humans

Quality control is a key link in the modernization process of traditional Chinese medicine(TCM). Studies have shown that the effects of active components in TCM depend on not only their chemical composition but also their suitable physical forms and states. The physical phase structures, such as micelles, vesicles, gels, and nanoparticles, can improve the solubility, delivery efficiency, and targeting precision of active components. These structures significantly enhance the pharmacological activity while reducing the toxicity and side effects, demonstrating functional activity surpassing that of active components and highlighting the key effects of "structures" on "functions" of active components. Taking the physical phase structure as a breakthrough point, this paper outlines the common types of TCM physical phase structures. Furthermore, this paper explores how to realize the quality upgrading of TCM through the precise regulation of physical phase structures based on the current applications and potential of TCM physical phase structures in processing to increase the efficacy and reduce the toxicity, compounding and decocting processes, drug delivery systems, and quality control, aiming to provide novel insights for the future quality control of TCM.
Quality Control ; Drugs, Chinese Herbal/standards* ; Medicine, Chinese Traditional/standards* ; Humans ; Drug Delivery Systems

Infectious keratitis (IK) is a leading cause of blindness worldwide, primarily resulting from improper contact lens use, trauma, and a compromised immune response. The pathogenic microorganisms responsible for IK include bacteria, fungi, viruses, and Acanthamoeba. This review examines standard therapeutic agents for treating IK, including broad-spectrum empiric antibiotics for bacterial keratitis (BK), antifungals such as voriconazole and natamycin for fungal infections, and antiviral nucleoside analogues for viral keratitis (VK). Additionally, this review discusses therapeutic agents, such as polyhexamethylene biguanide (PHMB), for the treatment of Acanthamoeba keratitis (AK). The review also addresses emerging drugs and the challenges associated with their clinical application, including anti-biofilm agents that combat drug resistance and nuclear factor kappa-B (NF-κB) pathway-targeted therapies to mitigate inflammation. Furthermore, methods of Photodynamic Antimicrobial Therapy (PDAT) are explored. This review underscores the importance of integrating novel and traditional therapies to tackle drug resistance and enhance drug delivery, with the goal of advancing treatment strategies for IK.