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 To investigate the role of magnesium ion(Mg2+)in cisplatin-induced acute kidney injury(Cis-AKI)in kidney organoids and HK-2 cells,as well as the potential mechanism.Methods Initially,we utilized human-derived induced pluripotent stem cells(iPSCs)to construct kidney organoids,and then built a Cis-AKI model based on kidney organoids.HE staining was used to observe the structure of kidney organoids,and immunofluorescence staining was used to observe the localization of markers and the expression of cleaved caspase-3.qRT-PCR was conducted to detect mRNA levels of tubular and glomerular markers,as well as inflammatory factors.Subsequently,the kidney organoids were randomly divided into control group,cisplatin group(Cis group),and Mg2+pretreatment group(Cis+Mg2+group).CCK-8 and ATP content assays were employed to evaluate the cell viability of renal tubular epithelial cells.TUNEL staining was performed to detect the apoptosis of renal tubular epithelial cells.Western blot was utilized to detect the expression of apoptosis-associated proteins(Bcl-2,Bax,cleaved caspase-3)and organic cation transporter 2(OCT2).Immunofluorescence was used to detect the localization and expression of OCT2.Results On the 10th day,the tubular structure in kidney organoids was visible,with abundant expression of renal markers.Treatment with 10 μmol/L cisplatin resulted in structural damage to kidney organoids,significantly increased expression of cleaved caspase-3 and mRNA levels of inflammatory factors,and significantly decreased ATP content.Compared with the Cis group,the Cis+Mg2+group showed increased ATP content in kidney organoids,reduced number of TUNEL-positive cells,significantly decreased expression of apoptosis-associated proteins,and significantly decreased expression of OCT2.However,there was no significant improvement in HK-2 cell viability,the number of TUNEL-positive cells,or apoptosis-associated proteins in the Cis+Mg2+group,and HK-2 cells did not express OCT2.Conclusion Kidney organoid is an ideal in vitro model to study the pathogenesis and treatment of Cis-AKI.Mg2+pretreatment can significantly reduce the damage of kidney organoids induced by cisplatin,and the mechanism may be related to the downregulation of OCT2.

Aim To study the electrophysiological mechanism of dopamine inhibiting insulin secretion hv voltage-dependent potassium ( Kv) channels.Methods Islets and (3 cells were isolated from male SD rats.D,-like receptor agonist ( SKP38393), D2-like receptor agonist (Quinpirole) and antagonist (Epiclopride) were used according to the experiment.Insulin secretion was detected by insulin radioimmunoassay.Whole-cell j J patch-clamp technique was applied to detect Kv channel currents and action potential duration of p cells.Di- BAC4(3) staining was used to observe membrane potential.Results SKF38393 did not affect insulin secretion and the Kv channel currents.Quinpirole signifi cantly inhibited insulin secretion and increased Kv channel currents.Dopamine significantly inhibited insulin secretion, increased Kv channel currents and shortened action potential duration of p cells, which could be reversed by epiclopride.In addition, dopamine de-creased membrane potential of INS-1 cells.Conclusions Dopamine inhibits insulin secretion by acting on D2-like receptors, resulting in actived Kv channels, shortened action potential duration and decreased cell membrane potential.

Lycii Cortex, the dry root bark of Lycium barbarum(Solanaceae), is rich in chemical compositions with unique structures, such as organic acids, lipids, alkaloids, cyclopeptides and other components, and plays an important role in traditional Chinese medicine. It has the effect of cooling blood and removing steam, clearing lung and reducing fire. It is mainly used in the treatment of hot flashes due to Yin deficiency, hectic fever with night sweat, cough, hemoptysis and internal heat and diabetes. Modern pharmacological studies have shown that the crude extract or monomer of Lycii Cortex has a variety of pharmacological activities, such as hypoglycemic, hypotensive, hypolipidemic, antibacterial, and antiviral effects. In this paper, the chemical constituents and pharmacological effects of Cortex Lycii were reviewed in order to further clarify its effective substances, promote the development of medical undertakings, and ensure the "Healthy China" plan.
China ; Hypoglycemic Agents ; Lycium ; Medicine, Chinese Traditional ; Plant Bark

Buffaloes ; Cells, Cultured ; In Vitro Techniques ; Primary Cell Culture ; Stem Cells ; Tretinoin

Currently, the systems for culturing buffalo spermatogonial stem cells (SSCs) in vitro are varied, and their effects are still inconclusive. In this study, we compared the effects of culture systems with undefined (foetal bovine serum) and defined (KnockOut Serum Replacement) materials on the in vitro culture of buffalo SSC-like cells. Significantly more DDX4- and UCHL1-positive cells (cultured for 2 days at passage 2) were observed in the defined materials culture system than in the undefined materials system (p < 0.01), and these cells were maintained for a longer period than those in the culture system with undefined materials (10 days vs. 6 days). Furthermore, NANOS2 (p < 0.05), DDX4 (p < 0.01) and UCHL1 (p < 0.05) were expressed at significantly higher levels in the culture system with defined materials than in that with undefined materials. Induction with retinoic acid was used to verify that the cultured cells maintained SSC characteristics, revealing an SCP3⁺ subset in the cells cultured in the defined materials system. The expression levels of Stra8 (p < 0.05) and Rec8 (p < 0.01) were significantly increased, and the expression levels of ZBTB16 (p < 0.01) and DDX4 (p < 0.05) were significantly decreased. These findings provided a clearer research platform for exploring the mechanism of buffalo SSCs in vitro.

Currently, the systems for culturing buffalo spermatogonial stem cells (SSCs) in vitro are varied, and their effects are still inconclusive. In this study, we compared the effects of culture systems with undefined (foetal bovine serum) and defined (KnockOut Serum Replacement) materials on the in vitro culture of buffalo SSC-like cells. Significantly more DDX4- and UCHL1-positive cells (cultured for 2 days at passage 2) were observed in the defined materials culture system than in the undefined materials system (p < 0.01), and these cells were maintained for a longer period than those in the culture system with undefined materials (10 days vs. 6 days). Furthermore, NANOS2 (p < 0.05), DDX4 (p < 0.01) and UCHL1 (p < 0.05) were expressed at significantly higher levels in the culture system with defined materials than in that with undefined materials. Induction with retinoic acid was used to verify that the cultured cells maintained SSC characteristics, revealing an SCP3⁺ subset in the cells cultured in the defined materials system. The expression levels of Stra8 (p < 0.05) and Rec8 (p < 0.01) were significantly increased, and the expression levels of ZBTB16 (p < 0.01) and DDX4 (p < 0.05) were significantly decreased. These findings provided a clearer research platform for exploring the mechanism of buffalo SSCs in vitro.

Objective To investigate the predictive value of extra pulmonary multiple factors including creatine kinase-isoenzyme MB (CK-MB) for the prognosis of patients with acute paraquat poisoning.Methods A retrospectively analysis were conducted on 641 patients who were treated at the First Affiliated Hospital of Zhengzhou University due to oral paraquat poisoning from October 2002 to April 2017.The observation end point was that the patients died from paraquat poisoning within 3 months after admission or were still alive within 3 months after paraquat poisoning.The patients' data were retrieved,including general information,the dose of poison,urinary paraquat concentration,arterial blood gas analysis,alanine transaminase (ALT),total bilirubin (TBIL),uric acid (UA),aspartate transaminase (AST),creatine kinase (CK),CK-MB,B type natriuretic peptide (BNP),lactic dehydrogenase (LDH),high sensitivity troponin T (hsTnT),C-reaction protein (CRP) and procaicitonin (PCT).According to the patient's prognosis within 3 months,the patients were divided into a survival group and a non-survival group.The above indicators were compared between the two groups and the diagnostic value of CK-MB for acute paraquat poisoning was analyzed according to the receiver operating characteristics (ROC) curve.Collect the last arterial blood gas analysis,and laboratory test results were analyzed by binary logistic regression analysis to determine the risk factors for death in patients with acute paraquat poisoning.Results Among the 641 patients with acute paraquat poisoning,315 (49.1％) patients survived and 326 (50.9％) died.Compared with the survival group,patients in the non-survival groupthere were older,had a shorter hospital stay,and had a higher oral paraquat dose and urinary paraquat concentration;Lac,TBIL,UA,AST,CK,CK-MB,BNP,LDH,CRP and PCT were higher,while blood gas analysis index were lower in the non-survival group (P＜0.05).Binary logistic regression analysis showed that the dose of paraquat,CK-MB and AST were closely related to the prognosis of patients with acute paraquat poisoning.The optimal cut-off value of ingestion dose,the first urinary paraquat concentration on admission and CK-MB in predicting the prognosis of patients with acute paraquat poisoning were 7 g (AUC=0.918,sensitivity 80.6％,specificity 87.5％,Yoden index 0.681,P＜0.01),5.16 μg/mL (AUC=0.879,sensitivity 93.8％,specificity 70.1％,Yoden index 0.639,P＜0.01),and 18.2 U/L (AUC=0.846,sensitivity 83.9％,specificity 71.9％,Yoden index 0.558,P＜0.01),respectively.Binary logistic regression analysis of the last biochemical indicators of paraquat poisoning showed that the dose of poison,the last CK-MB,the last SCr,urinary paraquat concentration,and the last blood Na+ were closely related to the prognosis of patients with acute paraquat poisoning.Among them,the last CK-MB＞18.05 U/L often indicated poor prognosis (AUC=0.808,sensitivity 79.7％,specificity 65.8％,Yoden index 0.455,P＜0.01).Conclusions In the treatment of patients with acute paraquat poisoning,there are significant differences in extra pulmonary factors such as heart,liver,kidney,electrolytes and inflammatory markers in patients with different prognosis,so the monitoring and follow-up should be improved,in addition to focusing on the presence and treatment of pulmonary fibrosis.In particular,CK-MB is an independent risk factor for the prognosis of acute paraquat poisoning.In the late stage of poisoning,CK-MB,SCr,and blood Na+ have a strong predictive value for the prognosis of the patients,and we should pay attention to the regular follow-up of the above mentioned laboratory items.

OBJECTIVE: To observe the clinical effect of platelet rich plasma (PRP) combined with β tricalcium phosphate bioceramic bone in the treatment of non traumatic necrosis of the femoral head in ARCO stageⅡ. METHODS: From January 2017 to December 2018, 100 patients (160 hips) with ARCO stageⅡnon traumatic necrosis of the femoral head were divided into PRP group and control group. In PRP group, 50 patients (80 hips), 22 males and 28 females, aged from 18 to 65 (43.47± 7.23) years, with a course of 4 to 18 (15.8±2.9) months, underwent core decompression and bone grafting combined with PRP implantation. There were 50 cases (80 hips) in the control group, including 27 males and 23 females, aged 20 to 63 (45.72± 7.43) years, and the course of disease was 6 to 19 (14.9±3.8) months. Hip X-ay film was followed up after operation. Harris score and VAS score were used to evaluate the curative effect, and the survival rate of hip joint was recorded. RESULTS: All patients had good wound healing, no infection, thrombosis and other complications. All patients were followed up for 12 to 14 (12.0±0.4) months. Twelve months after operation, the image expression of PRP group was better than that of control group( CONCLUSION Platelet-rich plasma(PRP) combined with artificialbone for core decompression and bone grafting can change the situation of simple artificial bone implantation and uncertain curative effect, improve the success rate of this operation, effectively reduce the collapse rate of femoral head necrosis in the early and middle stage, delay or even avoid hip replacement.
Adolescent ; Adult ; Aged ; Artemisinins ; Bone Transplantation ; Decompression, Surgical ; Female ; Femur Head/surgery* ; Femur Head Necrosis/surgery* ; Humans ; Male ; Middle Aged ; Naphthoquinones ; Platelet-Rich Plasma ; Treatment Outcome ; Young Adult

Objective To investigate the molecular characteristics of human immunodeficiency virus/acquired immune deficiency syndrome (HIV/AIDS) patients receiving antiretroviral therapy (ART) in Guizhou Province. Methods Using a convenience sampling strategy, 8 583 samples were collected in Guizhou and an investigation was conducted including face-to-face questionnaire interview and HIV testing. Results 1 511 cases failed in HIV suppression (viral load, VL>1 000 copies/ml). 1 410 cases (93.31%) were successfully genotyped with HIV pol gene, among which 51.42% were genotyped as CRF01_AE, 26.67% as CRF07_BC and 16.1% as CRF08_BC. Conclusion The subtype changes caused by HIV gene mutation should precede the changes of main transmission routes of HIV through the analysis in recent years. Timely monitoring the changes of HIV subtypes can be one of the main bases for the prevention and control of AIDS.