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.

AIM: To investigate the expression and clinical diagnostic value of toll-like receptor 4(TLR4)and nuclear factor-κB(NF-κB)in conjunctival epithelial cells and tears of patients with dry eye.METHODS: From January 2023 to June 2024, 104 dry eye patients(104 eyes, disease group)who visited our hospital and 100 healthy individuals(100 eyes, control group)who underwent physical examination were selected. The changes of TLR4 and NF-κB in conjunctival epithelial cells and tears were analyzed. Pearson analysis was applied to analyze the correlation between TLR4 and NF-κB expression in conjunctival epithelial cells and tears. Logistic analysis was applied to analyze the factors that affected dry eye. ROC was applied to analyze the diagnostic value of TLR4 and NF-κB expression in conjunctival epithelial cells and tears for dry eye.RESULTS: The differences in the use of eye drops, tear film break-up time(BUT), Schirmer's test(SⅠt), tear film thickness(TFT), and corneal fluorescein staining(CFS)scores between the disease group and the control group were statistically significant(all P<0.01). The expression levels of TLR4 and NF-κB in conjunctival epithelial cells and tears in the disease group were significantly higher than those in the control group(all P<0.01). There was a positive correlation between TLR4 and NF-κB in conjunctival epithelial cells and tears(r=0.392, 0.348, all P<0.05). Frequent use of eye drops, CFS score, TLR4, and NF-κB were risk factors for dry eye(OR=2.153, 3.183, 1.578, 2.452, all P<0.05), while BUT, SⅠt, and TFT were protective factors for dry eye(OR=0.654, 0.755, 0.276, all P<0.05). The sensitivity, specificity, and AUC of TLR4 combined with NF-κB in conjunctival epithelial cells in the diagnosis of dry eye were 86.54%, 81.00%, and 0.889, respectively. The combination of TLR4 and NF-κB had higher diagnostic value for dry eye than uncombined diagnosis(Zcombination-TLR4=3.506, P=0.001; Zcombination-NF-κB=3.165, P=0.002). The sensitivity, specificity, and AUC of TLR4 combined with NF-κB in tears for diagnosing dry eye were 82.69%, 70.00%, and 0.818, respectively. The combination of TLR4 and NF-κB in tears had higher diagnostic value for dry eye than uncombined diagnosis(Zcombination-TLR4=3.117, P=0.002; Zcombination-NF-κB=2.363, P=0.018).CONCLUSION: The expression levels of TLR4 and NF-κB in conjunctival epithelial cells and tears of patients with dry eye are elevated. TLR4 and NF-κB are related to the development of dry eye, and that elevated levels of both are associated with an increased risk of dry eye disease. The combination of TLR4 and NF-κB has a certain diagnostic significance for dry eye.

Tumors are the second leading cause of death worldwide. Exosomes are a type of extracellular vesicle secreted from multivesicular bodies, with particle sizes ranging from 40 to 160 nm. They regulate the tumor microenvironment, proliferation, and progression by transporting proteins, nucleic acids, and other biomolecules. Compared with other drug delivery systems, exosomes derived from different cells possess unique cellular tropism, enabling them to selectively target specific tissues and organs. This homing ability allows them to cross biological barriers that are otherwise difficult for conventional drug delivery systems to penetrate. Due to their biocompatibility and unique biological properties, exosomes can serve as drug delivery systems capable of loading various anti-tumor drugs. They can traverse biological barriers, evade immune responses, and specifically target tumor tissues, making them ideal carriers for anti-tumor therapeutics. This article systematically summarizes the methods for exosome isolation, including ultracentrifugation, ultrafiltration, size-exclusion chromatography (SEC), immunoaffinity capture, and microfluidics. However, these methods have certain limitations. A combination of multiple isolation techniques can improve isolation efficiency. For instance, combining ultrafiltration with SEC can achieve both high purity and high yield while reducing processing time. Exosome drug loading methods can be classified into post-loading and pre-loading approaches. Pre-loading is further categorized into active and passive loading. Active loading methods, including electroporation, sonication, extrusion, and freeze-thaw cycles, involve physical or chemical disruption of the exosome membrane to facilitate drug encapsulation. Passive loading relies on drug concentration gradients or hydrophobic interactions between drugs and exosomes for encapsulation. Pre-loading strategies also include genetic engineering and co-incubation methods. Additionally, we review approaches to enhance the targeting, retention, and permeability of exosomes. Genetic engineering and chemical modifications can improve their tumor-targeting capabilities. Magnetic fields can also be employed to promote the accumulation of exosomes at tumor sites. Retention time can be prolonged by inhibiting monocyte-mediated clearance or by combining exosomes with hydrogels. Engineered exosomes can also reshape the tumor microenvironment to enhance permeability. This review further discusses the current applications of exosomes in delivering various anti-tumor drugs. Specifically, exosomes can encapsulate chemotherapeutic agents such as paclitaxel to reduce side effects and increase drug concentration within tumor tissues. For instance, exosomes loaded with doxorubicin can mitigate cardiotoxicity and minimize adverse effects on healthy tissues. Furthermore, exosomes can encapsulate proteins to enhance protein stability and bioavailability or carry immunogenic cell death inducers for tumor vaccines. In addition to these applications, exosomes can deliver nucleic acids such as siRNA and miRNA to regulate gene expression, inhibit tumor proliferation, and suppress invasion. Beyond their therapeutic applications, exosomes also serve as tumor biomarkers for early cancer diagnosis. The detection of exosomal miRNA can improve the sensitivity and specificity of diagnosing prostate and pancreatic cancers. Despite their promising potential as drug delivery systems, challenges remain in the standardization and large-scale production of exosomes. This article explores the future development of engineered exosomes for targeted tumor therapy. Plant-derived exosomes hold potential due to their superior biocompatibility, lower toxicity, and abundant availability. Furthermore, the integration of exosomes with artificial intelligence may offer novel applications in diagnostics, therapeutics, and personalized medicine.

OBJECTIVE:External fixators and plate internal fixation are commonly used treatments for comminuted distal radius fractures,each with its own advantages and disadvantages in clinical practice. To systematically evaluate the clinical efficacy and safety of external fixator and plate internal fixation in the treatment of comminuted distal radius fractures,and to provide a basis for the development of guidelines for the diagnosis and treatment of distal radius fractures with integrated traditional Chinese and Western medicine. METHODS:PubMed,Web of Science,Embase,Cochrane Library,China National Knowledge Infrastructure,China Biomedical Literature Database,VIP,andWanFang databases were systematically searched to include randomized controlled trials on external fixators and internal plate fixation for the treatment of comminuted distal radial fractures published from October 2013 to October 2023. The literature was screened according to the inclusion and exclusion criteria. Review Manager was used for literature quality evaluation and meta-analysis.RESULTS:(1) Eight articles were included,including 4 in Chinese and 4 in English,with a total sample size of 648 cases,including 328 cases in the external fixation stent group and 320 cases in the internal fixation plate group. (2) At 3 months after operation,the internal fixation plate group was superior to the external fixation stent group in the range of dorsal extension,palmar flexion and supination. At 12 months after operation,the grip strength,palmar inclination,palmar flexion,pronation and supination in the internal fixation plate group were better than those in the external fixation stent group. The postoperative infection in the internal fixation plate group was better than that in the external fixation stent group,and there was no statistical difference in other outcome indicators.CONCLUSION:Eight evidences showed that in the choice of treatment for comminuted distal radius fracture,both external fixation stent and incision plate internal fixation had good therapeutic effect,and plate internal fixation was better than other factors. However,for some special patients with highly severe comminuted distal radius fractures,poor bone quality,severely contaminated open fractures,and soft tissue swelling that did not allow incision surgery,external fixation was the first choice. The results of this study have limitations,and more high-quality,large-sample,multi-center randomized controlled trials are needed in the future,emphasizing the observation of long-term efficacy and other secondary indicators,and supplementing and optimizing the current research results.

Objective To explore the clinical efficacy of transcranial combined peripheral magnetic stimu-lation on bladder and voiding ability in patients with NB after spinal cord injury.Methods From September 2023 to October 2024,60 patients with NB after spinal cord injury were chosen from the Department of Rehabilitation Medicine of the First Affiliated Hospital of Bengbu Medical University,and were separated into the regular group(n=15),M1 area stimulation group(n=15),sacral nerve root stimulation group(n=15),and combined stimulation group(n=15).The control group underwent conventional rehabilitation treatment,the M1 area stimu-lation group added repetitive transcranial magnetic stimulation(rTMS)of the M1 area of the motor cortex,the sacral nerve root stimulation group added repetitive peripheral magnetic stimulation(rPMS)of the sacral 3 nerve roots,and the co-stimulation group added rTMS of the M1 area and rPMS of the sacral 3 nerve roots,and the treat-ment was carried out for 4 weeks in total.Urodynamic indices,voiding diaries,and neurogenic bladder symptom scores(NBSS)were assessed before and after therapy in the four groups.Results Before therapy,the differences in maximum bladder capacity,bladder capacity at the time of the first urge to urinate,maximum urine flow rate,residual urine volume,average number of urination per day,average daily urinary output,and NBSS scores of the four groups were not statistically significant when compared with those of the pre-treatment group(P<0.05);after treatment,the maximum bladder capacity,bladder capacity at the time of the first urge to urinate,maximum urine flow rate,residual urine volume,average number of urination per day,average daily urinary output,and NBSS scores were all improved compared with those before treatment(P<0.05);in comparison between the groups,after treatment,the Urodynamic indices,voiding diaries,and NBSS scores of the combined group were better than those of the other three groups(P<0.05).Conclusion Transcranial combined peripheral bidirectional feedback magnetic stimulation therapy can improve urodynamic and voiding symptoms and promote the recovery of bladder and voiding function in patients with NB after spinal cord injury

Objective:To develop an early exercise rehabilitation nursing for elderly patients undergoing total hip arthroplasty (THA) based on the best evidence, and explore its clinical effects.Methods:This study was a randomized controlled trial. From August to December 2022, convenience sampling was used to select 126 elderly THA patients admitted to the Department of Orthopedics of the First Affiliated Hospital of Naval Medical University as participants. Patients were divided into observation group and control group using a random number table method, with 63 cases in each group. Control group was treated with a usual exercise rehabilitation nursing, while observation group was treated with an early exercise rehabilitation nursing for elderly THA patients based on the best evidence. Changhai Pain Rating Scale, Harris Hip Score, and Barthel Index were used to evaluate pain, hip function, and activities of daily living in two groups of patients. The first time off the bed, hospital stay, and incidence of complications were recorded and compared between the two groups.Results:The Harris Hip scores of observation group patients three days after surgery and on the day of discharge were (61.13±4.82) and (77.84±4.43), respectively, which were higher than those of control group (50.75±6.19) and (70.25±7.47), and the differences were statistically significant ( t=10.502, 6.937; both P<0.01). Barthel Index scores of observation group patients were higher than those of control group on the day of surgery, three days after surgery, and the day of discharge, and the differences were statistically significant ( t=3.531, 4.609, 10.112; all P<0.01). The incidence of postoperative complications in observation group was 11.11% (7/63), while in control group it was 42.86% (27/63), with a statistically significant difference (χ 2=16.113, P<0.01). There were no statistically significant differences in the resting pain score and exercise pain score between the two groups of patients on the day after surgery ( t=-1.693, -1.354; both P>0.05). The resting pain score and exercise pain score of observation group were lower than those of control group three days after surgery and on the day of discharge, and the differences were statistically significant (all P<0.05). Observation group had a shorter first time off the bed and hospital stay (1.34±0.37) and (5.48±0.66) days compared to control group (2.13±0.48) and (6.29±0.83) days, with statistically significant differences ( t=-10.346, -6.063; both P<0.01) . Conclusions:An evidence-based early exercise rehabilitation nursing for elderly THA patients can reduce the degree of pain, minimize the occurrence of complications, shorten the first time off the bed and hospital stay, and promote recovery of elderly patient.

OBJECTIVE: To explore the influencing factors on the prognosis of patients with multiple myeloma (MM) after bortezomib treatment, and construct a decision tree risk prediction model based on the influencing factors. METHODS: One hundred and seventy MM patients admitted to the People's Hospital of Jianyang City from January 2019 to June 2022 were selected as research subjects, and divided into poor prognosis group and good prognosis group according to the prognosis after bortezomib treatment. The clinical data of the patients were analyzed, univariate and logistic regression analysis were used to screen influencing factors, SPSS Modeler software was used to construct a decision tree prediction model, and the diagnostic performance of the decision tree risk prediction model was analyzed. RESULTS: The incidence of poor prognosis in 170 MM patients after bortezomib-based chemotherapy was 21.18%. Kappa light chain level≥19.4 mg/L, platelet count (PLT) ≤100×10⁹/L, homocysteine (Hcy) >22 μmol/L, serum creatinine (Scr) ≥176 μmol/L, lactate dehydrogenase (LDH) ≥300 U/L, serum ferritin (SF) >500 mg/L, and β₂-microglobulin (MG) >6 μg/L were independent risk factors for poor prognosis in MM patients after bortezomib treatment (all P < 0.05). The decision tree model selected 7 explanatory variables (Kappa light chain level, LDH, PLT, SF, β₂-MG, Scr, and Hcy) as nodes of the model, among which Kappa light chain level was the most important predictor. In addition, the area under the ROC curve (AUC) values of the decision tree model and logistic regression model were 0.895 and 0.881, respectively. The prediction performance of the decision tree model was better than that of the logistic regression model ( Z=3.325, P =0.005). CONCLUSION The decision tree model has high value in predicting the prognosis after bortezomib treatment in MM patients, which can screen high-risk factors that affect poor prognosis, providing practical references for clinical healthcare professionals to take preventive treatment for high-risk MM patients.
Humans ; Bortezomib/therapeutic use* ; Multiple Myeloma/diagnosis* ; Decision Trees ; Prognosis ; Algorithms ; Risk Factors ; Male ; Female ; Middle Aged

OBJECTIVE: To investigate the effectiveness of Xuanshen Yishen Decoction (XYD) in the treatment of hypertension. METHODS: Hospital electronic medical records from 2019-2023 were utilized to emulate a randomized pragmatic clinical trial. Hypertensive participants were eligible if they were aged ⩾40 years with baseline systolic blood pressure (BP) ⩾140 mm Hg. Patients treated with XYD plus antihypertensive regimen were assigned to the treatment group, whereas those who followed only antihypertensive regimen were assigned to the control group. The primary outcome assessed was the attainment rate of intensive BP control at discharge, with the secondary outcome focusing on the 6-month all-cause readmission rate. RESULTS: The study included 3,302 patients, comprising 2,943 individuals in the control group and 359 in the treatment group. Compared with the control group, a higher proportion in the treatment group achieved the target BP for intensive BP control [8.09% vs. 17.5%; odds ratio (OR)=2.29, 95% confidence interval (CI)=1.68 to 3.13; P<0.001], particularly in individuals with high homocysteine levels (OR=3.13; 95% CI=1.72 to 5.71; P<0.001; P for interaction=0.041). Furthermore, the 6-month all-cause readmission rate in the treatment group was lower than in the control group (hazard ratio=0.58; 95% CI=0.36 to 0.91; P=0.019), and the robustness of the results was confirmed by sensitivity analyse. CONCLUSIONS XYD could be a complementary therapy for intensive BP control. Our study offers real-world evidence and guides the choice of complementary and alternative therapies. (Registration No. ChiCTR2400086589).
Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Antihypertensive Agents/pharmacology* ; Blood Pressure/drug effects* ; Drugs, Chinese Herbal/pharmacology* ; Hypertension/physiopathology* ; Patient Readmission ; Treatment Outcome

OBJECTIVE: To explore the mechanism of Dihuang Yinzi (DHYZ) in the treatment of Alzheimer's disease (AD) by integrating metabolomics and experimental verification. METHODS: Forty-eight male APP/PS1 mice were divided into model, high- (DHYZ-H), medium- (DHYZ-M), and low-dose DHYZ (DHYZ-L) groups (12 mice per group) according to a random number table. Mice in DHYZ groups were gavaged with DHYZ 6.34, 12.68, and 25.35 g/(kg·d), respectively. Twelve C57BL/6 mice were gavaged with distilled water as the blank group. Metabolomics was used to analyze differential metabolites in the brains of mice. Morris water maze test was used to detect the memory abilities of mice. The hematoxylin-eosin staining and transmission electron microscopy were used to observe the general morphology and ultrastructure of neurons. The enzyme-linked immunosorbent assay was used to detect the levels of superoxide dismutase (SOD), reactive oxygen species (ROS), and amyloid β -protein 1-42 (A β_1-42). The real-time quantitative polymerase chain reaction was used to detect the mRNA expressions of density-regulated protein 1 (DRP1), fission 1 (FIS1), mitofusin-1 (MFN1), and optic atrophy protein 1 (OPA1). Western blot was used to detect the protein expressions of cyclic adenosine monophosphate (cAMP), protein kinase A (PKA), cAMP response binding protein (CREB), brain-derived neurotrophic factor (BDNF), synapsin 1 (SYN1), synaptophysin (SYP), and postsynaptic density protein 95 (PSD95). RESULTS: A total of 82 differential metabolites were identified in the brains of APP/PS1 mice, among which 7 differential metabolites could be regulated by DHYZ. After DHYZ intervention, the memory abilities of mice significantly increased (P<0.05 or P<0.01), the number of synapses and neurons in the hippocampus increased, and the mitochondrial morphology and structure were relatively intact. The DHYZ groups exhibited a significant reduction in hippocampal ROS and A β_1-42 levels, along with a significant elevation in SOD level (P<0.05 or P<0.01). The mRNA expressions of DRP1 and FIS1 were reduced, while the mRNA expressions of MFN1 and OPA1 were increased after DHYZ treatment (P<0.05 or P<0.01). The cAMP/PKA/CREB-BDNF pathway was activated, and the expressions of SYN1, SYP and PSD95 proteins were significantly increased in the DHYZ-H group (P<0.05 or P<0.01). CONCLUSIONS DHYZ could improve mitochondrial dynamics and synaptic plasticity in APP/PS1 mice, inhibit oxidative stress, and thereby enhancing learning and memory abilities in APP/PS1 mice. Its mechanism might be related to activation of the cAMP/PKA/CREB-BDNF signaling pathway.
Animals ; Brain-Derived Neurotrophic Factor/metabolism* ; Male ; Cyclic AMP Response Element-Binding Protein/metabolism* ; Brain/drug effects* ; Metabolomics ; Mice, Inbred C57BL ; Neuronal Plasticity/drug effects* ; Drugs, Chinese Herbal/therapeutic use* ; Cyclic AMP-Dependent Protein Kinases/metabolism* ; Cyclic AMP/metabolism* ; Reactive Oxygen Species/metabolism* ; Amyloid beta-Protein Precursor/metabolism* ; Mice, Transgenic ; Mice ; Amyloid beta-Peptides/metabolism* ; Signal Transduction/drug effects* ; Alzheimer Disease/drug therapy* ; Superoxide Dismutase/metabolism*

Human self-organizing cardioids, a recent breakthrough in cardiac organoid research, are constructed with the specialized cardiac lineage cells derived from human pluripotent stem cells (hPSCs) and have made rapid advancements since 2021. A key advantage of these organoids is their minimal reliance on external interventions, allowing them to more accurately replicate the heart's developmental processes through intrinsic signaling pathways, thereby closely mimicking natural cardiac characteristics. Consequently, they hold significant promise for improving drug safety evaluations, treating both congenital and acquired heart diseases, advancing eugenics practices, developing humanized cardiac disease models, conducting research in regenerative medicine, and understanding how unique environments (such as aerospace) affect human health. This review systematically describes the current various self-organizing cardioid construction techniques, comparing the structural differences caused by diverse signal stimulations, which would be instrumental in optimizing designs for more advanced and mature cardioids. Additionally, we summarize existing applications and address the challenges faced. Despite some uncertainties and challenges in current technologies and applications, this emerging cardiac organoid technology holds promise to provide new possibilities for cardiovascular medicine through continuous refinement.