Objective: To identify biomolecular markers closely related to the occurrence of kidney renal clear cell carcinoma (KIRC) and verify their expression levels in clinical samples. Methods: Stage Ⅰ KIRC mRNA sequencing data were obtained from The Cancer Genome Atlas (TCGA).Principal component analysis (PCA) was used for dimensionality reduction to screen differentially expressed genes (DEGs)，which then underwent GO and KEGG analyses.Weighted gene co-expression network analysis (WGCNA) was used to screen genes significantly related to KIRC，and a protein-protein interaction (PPI) network was constructed to screen hub genes.The diagnostic value of hub genes was evaluated with receiver operating characteristic (ROC) curve，and their prognostic value was analyzed using survival curve plots.The correlation between the mRNA expressions of hub genes and the pathological stages of KIRC was analyzed.Clinical samples of 20 patients with stage Ⅰ KIRC treated in our hospital were included，and the expressions of the hub genes in cancerous and adjacent tissues were detected with reverse transcription real-time quantitative polymerase chain reaction (RT-qPCR)，Western blotting，and enzyme-linked immunosorbent assay (ELISA). Results: A total of 8223 DEGs were screened out，including 4092 up-regulated ones and 4131 down-regulated ones.GO analysis showed that DEGs were related to bioadhesion，plasma membrane composition，and transporter activity.KEGG analysis showed that DEGs were related to pathways such as cell adhesion molecules，cytokine-cytokine receptor interactions，and interactions between viral proteins and cytokines and cytokine receptors.WGCNA analysis obtained 171 genes that were significantly related to stage Ⅰ KIRC.The hub gene，lymphocyte cytosolic protein 2 (LCP2)，screened out by the PPI network，was significantly related to stage Ⅰ KIRC.The area under the ROC curve was 0.96.The expression level was negatively correlated with the overall survival rate of patients.The expression of LCP2 was related to the stage and lymph node metastasis.Clinical verification showed that the mRNA and protein relative expressions of LCP2 in KIRC tissues were significantly higher than those in adjacent tissues (P<0.000 1). Conclusion: LCP2 is significantly up-regulated in stage Ⅰ KIRC tissues and can be used as a potential biomarker for the early diagnosis and treatment of KIRC.

Objective To investigate the epidemiological characteristics of dengue fever in Shenzhen City in 2024, so as to provide insights into formulation of the preventive and control measures for dengue fever. Methods The epidemiological data of dengue cases reported in Shenzhen City in 2024 were extracted from the China Disease Prevention and Control Information System and field epidemiological survey data of dengue fever in Shenzhen City, and the temporal, regional and population distributions of dengue fever cases, source of acquire dengue virus infections, disease diagnosis and treatment and outbreaks were analyzed. The dengue virus nucleic acid was tested and the serotypes of dengue virus were characterized using real-time quantitative reverse transcription PCR (RT-qPCR) assay, and the dengue virus gene was sequenced using next-generation sequencing (NGS). In addition, the surveillance on the density of Aedes albopictus was performed using Breteau index (BI) and mosquito oviposition index (MOI). Results A total of 1 735 dengue fever cases were reported in Shenzhen City in 2024, including 952 local cases and 783 imported cases. Most imported dengue fever cases acquired infections from eight cities of Foshan, Guangzhou, Zhongshan, Jiangmen, Dongguan, Zhaoqing, Huizhou, and Zhuhai in the Pearl River Delta region (664 cases, 84.8% of total imported cases) into Baoan, Longgang, and Nanshan districts. The epidemic exhibited an early onset and rapid progression, peaking during the period between September and November (1 632 cases, 94.1% of total cases), and dengue fever cases were distributed across 73 subdistricts in 10 districts, with most cases reported in densely populated central and western regions. The dengue fever cases had a male-to-female ratio of 1.9∶1.0, and a median age of 37 (21) years, with a higher median age among local cases than among imported cases [40 (20) years vs. 33(15) years; Z = -10.30, P < 0.05]. Housework, unemployment, workers, and business service were predominant occupations (1 405 cases, 81.0% of total cases), and there was a significant difference in the constituent ratio of occupations between local and imported cases (χ² = 92.30, P < 0.05). Among the 1 735 dengue fever cases, the median duration from onset to definitive diagnosis was 3.3 (2.9) days, and 1 686 cases (97.2%) were identified in healthcare facilities, with a low rate of hospitalization and isolation seen in 1 701 inpatients with available epidemiological data (485 cases, 28.5% of total inpatients). A total of 29 outbreaks of dengue fever occurred in Shenzhen City across 2024, which primarily in construction sites (27 outbreaks, 93.1% of total). Dengue virus type I was the dominant serotype causing dengue fever in Shenzhen City in 2024. Sequencing showed that the genomes of dengue virus from multiple dengue fever cases in Shenzhen City shared a high sequence homology with those from cities neighboring Shenzhen City, and there might be intra-city transmission of dengue virus among multiple construction sites in Shenzhen City. The Aedes albopictus density was significantly higher in Shenzhen City in 2024 than in 2023, peaking from May to September. The annual MOI values ranged from 0.9 to 14.0, and the BI values ranged from 0.6 to 6.0. Conclusions The overall epidemic of dengue fever was severe in Shenzhen City in 2024, which was greatly affected by case importation from neighboring cities, construction sites-centered local transmission, and the effectives of routine mosquito vector control was not satisfactory. Integrated dengue fever control measures should be implemented, focusing on regional joint prevention and control mechanisms, capacity building for mosquito vector control, addressing challenges in epidemic containment at construction sites, and strengthening case detection and management systems.

OBJECTIVES: To investigate the impact of prenatal fear stress on placental amino acid transport and emotion and cognition development in offspring rats. METHODS: Thirty pregnant Wistar rats were randomized equally into control and fear stress (induced using an observational foot shock model) groups. In each group, placental and serum samples were collected from 6 dams on gestational day 20, and the remaining rats delivered naturally and the offspring rats were raised under the same conditions until 8 weeks of age. Emotional and cognitive outcomes of the offspring rats were assessed with behavioral tests, and placental structure was examined using HE staining. Bioinformatics analysis was used to identify differentially expressed placental transporter genes under fear stress. The expressions of system A and system L amino acid transporters, along with other specialized transporters, were detected using qRT-PCR and Western blotting. Fetal serum amino acid concentrations were determined by HPLC. The correlations between fetal amino acid levels and behavioral outcomes of the offspring rats were analyzed. RESULTS: The dams with fear stress showed reduced open-field activity and increased freezing behavior with significantly decreased placental weight, fetal weight, and fetal-to-placental ratio. Bioinformatics analysis revealed 28 differentially expressed transporter genes involved mainly in amino acid transport. In the fear stress group, fetal serum amino acid levels were significantly lowered and Slc38a1, Slc43a1, Slc43a2, Slc7a8, Slc6a6, Slc1a1 and Slc6a9 mRNA and protein expressions were all downregulated. The offspring rats in fear stress group exhibited decreased novel object preference and spontaneous alternation with reduced open arm exploration and increased immobility in emotional tests. Lower early-life amino acid levels was found to correlate with impaired adult cognition. CONCLUSIONS Prenatal fear stress in rats impairs placental amino acid transporter expression and reduces fetal serum amino acid levels, potentially contributing to long-term cognitive deficits in the offspring rats.
Animals ; Female ; Pregnancy ; Placenta/metabolism* ; Fear ; Rats ; Rats, Wistar ; Cognition ; Prenatal Exposure Delayed Effects ; Stress, Psychological ; Amino Acids/blood* ; Amino Acid Transport Systems/metabolism*

CD20 is a surface marker protein of B-cell lymphoma, and its extracellular region is the target of specific antibodies and drugs. To obtain a cheap and easily modified specific preparation targeting CD20, we optimized the gene of CD20 extracellular region according to codon degeneracy to facilitate its expression in Escherichia coli. The optimized gene was cloned into pGEX-4T-1 vector, and the recombinant vector was transformed into E. coli BL21(DE3) for expression. The purified protein was identified by SDS-PAGE and Western blotting. Systematic evolution of ligands by exponential enrichment (SELEX) was employed to screen the ssDNA aptamer that specifically binds to the fusion protein, and the affinity of the aptamer to CD20 was detected by flow cytometry. Then, the cytotoxicity test was carried out to examine the inhibitory effect of the aptamer on B lymphoma cells. In this study, we established the prokaryotic expression method of CD20 and obtained the aptamer specifically binding to the extracellular region of CD20, which laid a foundation for the development of therapeutic drugs targeting CD20.
Humans ; Escherichia coli/metabolism* ; SELEX Aptamer Technique/methods* ; Aptamers, Nucleotide/genetics* ; Antigens, CD20/metabolism* ; Ligands

OBJECTIVE To provide standardized whole-process guidance on drug traceability codes for medical institutions in Sichuan province， ensuring medication safety and compliance with medical insurance supervision requirements. METHODS Based on evidence-based principles and expert consensus， Expert Consensus on Whole-process Management of Drug Traceability Codes in Medical Institutions of Sichuan Province （hereinafter referred to as the Consensus） was formulated through systematic literature review， field investigations， establishment of a multidisciplinary expert committee and multiple rounds of questionnare consultation via the modified Delphi method， and finalized through consensus meetings. RESULTS & CONCLUSIONS The Consensus clarifies key operating procedures for code verification， code assignment and code return， whole-process operational standards for drug warehouse acceptance and storage， drug warehouse outbound delivery and pharmacy acceptance check， drug distribution and dispensing in pharmacy and intravenous admixture center， medication administration in nursing units and examination departments， as well as drug return process. Key recommendations are proposed such as improving the core functions of the drug traceability system， unifying the hospital-wide traceability code database， strengthening the management of traceability codes for backup medications， establishing a management organization and institutional framework， and optimizing the architectural design and data governance requirements of the drug traceability system. The release of the Consensus will provide scientific， standardized and implementable practical guidelines for medical institutions of Sichuan province， helping to improve closed-loop management of the drug traceability system， strengthen medication safety and fulfil medical insurance fund supervision.

Objective To investigate the correlation between the expression level of absent in melanoma 2(AIM2)in serum and the severity of acute cerebral infarction(ACI),and to evaluate the value of AIM2 in predicting early neurological deterioration(END)after intravenous thrombolysis with alteplase in ACI patients.Methods A total of 150 ACI patients were enrolled in ACI group,and an-other 30 healthy individuals physical examination were selected as control group.ACI patients were further divided into mild,moderate and severe groups according to the National Institutes of Health Stroke Scale(NIHSS)score at admission,and they were also divided into END and non-END groups based on the occurrence of END after intravenous thrombolysis.Clinical materials of ACI patients were collected;the expression level of serum AIM2 was measured by enzyme-linked immunosorbent assay(ELISA);the Logistic regression analysis was performed to identify the factors influencing END after intravenous thrombolysis in ACI patients;the receiver operating characteristic(ROC)curve was plot-ted to analyze the clinical performance of serum AIM2 expression level in predicting END after intravenous thrombolysis in ACI patients.Results The serum AIM2 expression level in the ACI group and the control group was(58.29±5.97)and(36.81±3.03)ng/mL respectively,with a significant be-tween-group difference(t=43.23,P＜0.05).The serum AIM2 expression level gradually in-creased in the mild,moderate and severe groups,with significant differences between each pair of groups(P＜0.01).The serum AIM2 expression level in ACI patients was positively correlated with the NIHSS score(r=0.941,P＜0.01).The NIHSS score,time from admission to thrombolysis,low-density lipoprotein cholesterol(LDL-C),C-reactive protein,and AIM2 level in the END group were significantly higher or longer than those in the non-END group(P＜0.05).The results of mul-tivariate Logistic regression analysis showed that NIHSS score(OR=3.871,P＜0.001),time from admission to thrombolysis(OR=2.885,P=0.002),LDL-C(OR=3.118,P＜0.001)and AIM2(OR=3.761,P＜0.001)were influencing factors for END after intravenous thrombolysis in ACI patients.ROC curve showed that the area under the curve(AUC)for predicting END after in-travenous thrombolysis based on the serum AIM2 expression level at admission in ACI patients was 0.911;when the cut-off value of AIM2 was 66.56 ng/mL,the diagnostic sensitivity and specificity were 91.23％and 90.15％,respectively.Conclusion The expression level of serum AIM2 is sig-nificantly increased in ACI patients,and AIM2 expression level has certain advantages in predicting END after intravenous thrombolysis in ACI patients.

Objective:To explore the protective effect of small ubiquitin-related moditier protein specific peptidase 2 (SENP2) on acute kidney injury in septic mice by regulating NLRP3 inflammasome activation.Methods:Forty C57BL/6 mice were divided into 4 groups: namely the wild-type sham group (WT-Sham), wild-type cecal ligation and perforation group (WT-CLP), SENP2 gene knockout sham group (KO-Sham), and SENP2 gene knockout cecal ligation and perforation group (KO-CLP), with 10 mice in each group. It was observed the pathological damage of kidney tissue in each group of mice, used ELISA method to detect blood creatinine (SCr), blood urea nitrogen (BUN), plasma neutrophil gelatinase related lipoprotein (pNGAL), and plasma kidney injury molecule 1 (pKIM-1) levels, and determined plasma TNF-α, IL-1β, IL-4 and IL-10 levels. It was used immunohistochemical method to detect the expression of SENP2 protein in renal tissue, while used western-blotting to detect NLRP3, IL-1β, Caspase-1 and ASC protein expression in renal tissue.Results:Compared with the WT-Sham and KO-Sham groups, in the WT-CLP and KO-CLP groups, the pathological damage scores of the kidney tissue and the levels of SCr, BUN, pNGAL, pKIM-1, TNF-α and IL-1β in plasma were all significantly increased (all P<0.001), while the levels of IL-4 and IL-10 were all significantly reduced (all P<0.001), and the levels of NLRP3, IL-1β, Caspase-1 and ASC proteins were all significantly increased (all P<0.001). Moreover, compared with the WT-CLP group, the KO-CLP group showed a significant decrease in renal tissue injury scores in mice ( P?0.01), the levels of SCr, BUN, pNGAL, pKIM-1, TNF-α and IL-1β were all significantly reduced (all P<0.05), and the levels of NLRP3 [(0.71 ± 0.04) vs. (0.89 ± 0.01), P=0.011], IL-1β [(0.41 ± 0.02) vs. (0.57 ± 0.01), P=0.004], Caspase-1 [(0.41 ± 0.02) vs. (0.56 ± 0.02), P=0.009], and ASC [(0.27 ± 0.01) vs. (0.41 ± 0.02), P=0.009] were all significantly reduced. Conclusion:SENP2 participates in the occurrence and development of septic AKI by regulating the activation of NLRP3 inflammasomes.

BACKGROUND:Bone grafting is one of the important steps in the treatment of thoracolumbar burst fractures.Because the fracture involves the spinal canal or is accompanied by spinal cord nerve damage,severe fracture bleeding and other factors,minimally invasive bone grafting for thoracolumbar burst fractures is restricted.At present,the minimally invasive treatment of thoracolumbar burst fractures is limited to percutaneous screw fixation under the tunnel.Minimally invasive percutaneous bone grafting of injured vertebrae is rarely reported,and percutaneous precise bone grafting under the endplate has not yet been reported. OBJECTIVE:To investigate the clinical effect of subcutaneous endplate bone graft support reduction combined with percutaneous pedicle screw short-segment fixation in the treatment of A3+B2 thoracolumbar burst fractures. METHODS:From June 2017 to December 2021,90 patients with A3+B2 type asymptomatic thoracolumbar burst fracture were randomly divided into 3 groups according to admission time.In group A,33 patients received the bone graft funnel accurately placed through the pedicle channel by percutaneous puncture under C-arm fluoroscopy,bone graft support reduction under the fracture endplate,percutaneous pedicle screw fixation.In group B,30 patients received multifissure intermuscular approach through pedicle bone graft support reduction combined with pedicle screw fixation.In group C,27 patients received percutaneous pedicle screw short-segment fixation under postural reduction.All patients were followed up for at least 18 months after surgery.The clinical data of the three groups,including preoperative,postoperative and last follow-up Cobb angle,anterior edge height ratio and visual analog scale pain score,were compared and analyzed. RESULTS AND CONCLUSION:(1)There were no significant differences in age,sex,injury segment and causative factors among the three groups(P>0.05).(2)All patients at follow-up had no neurological impairment,no obvious lumbar posterior deformity or intractable low back pain.(3)The operation time of group C was less than that of group A and group B(P<0.05).Intraoperative blood loss was less in group A and group C than in group B(P<0.05).(4)There were no significant differences in the anterior edge height ratio and Cobb angle among the three groups(P>0.05).Postoperative data in groups A and B were better than that in group C.At last follow-up,group A and group B outperformed group C(P<0.05).The height and Cobb angle of the vertebral body lost in the three groups were smaller in groups A and B than those in group C(P<0.05).(5)Visual analog scale pain score was better in groups A and C than that in group B after surgery(P<0.05).There was no significant difference in visual analog scale pain score among the three groups at last follow-up(P>0.05).(6)In group C,there was one case of loose internal fixation and displacement in 1 month after surgery,and the vertebral height was lost again with back pain,and after strict bed rest for 6 weeks,the vertebral height loss was not aggravated,the pain was relieved,and the internal fixation was removed after 1 year,and the height loss at the last follow-up was not aggravated.There were no cases of failure of internal fixation in groups A and B.(7)It is indicated that subcutaneous endplate bone graft support reduction combined with percutaneous pedicle screw short-segment fixation in the treatment of A3+B2 thoracolumbar burst fracture has the advantages of less trauma,less bleeding and light postoperative pain symptoms,and the effect of injury vertebral reduction and height maintenance is the same as the reduction through pedicle bone grafting support and short segment fixation with pedicle screws through the multifidus space approach.

Objective To investigate the dynamic changes and molecular mechanisms of myocardial injury and autophagy as the body's self-protective mechanism at different time points after cerebral ischemia/reperfusion(CI/R).Methods A CI/R model was established in male Sprague-Dawley rats using the Longa thread method.Rats that underwent CI/R following middle cerebral artery occlusion were classified into 6-,12-,24,and 48-hour groups according to the reperfusion time.The concentrations of reactive oxygen species and reac-tive nitrogen species were measured to evaluate myocardial oxidative stress.Cardiomyocyte apoptosis and autophagosomes were observed in the myocardium.Additionally,dynamic changes in myocardial autophagy,autophagic flux,and protein and gene expression of auto-phagy regulators were detected.Results Oxidative-stress-induced injury and apoptosis were observed in the myocardium after CI/R.The number of autophagosomes in cardiomyocytes increased,peaking in the 12 h group,and autophagic flux was impaired during the first 12 h after CI/R.Beclin 1,mammalian target of rapamycin(mTOR),and adenosine monophosphate-activated protein kinase(AMPK)expression levels in the myocardium increased during the first 48 h after CI/R,peaking in the 12 h group.This was consistent with changes in autophagy,which showed significant differences compared with the control group.Conclusion These results indicated that autophagy plays a protective role against CI/R-induced myocardial injury.Furthermore,Beclin 1-mediated autophagy/apoptosis and mTOR-mediated autophagy mutual feedback pathways play important roles in the regulation of autophagy.

Myocardial ischemia-reperfusion injury （MIRI） is a common injury in the treatment of ischemic heart diseases. MIRI can be categorized as chest impediment and palpitation in traditional Chinese medicine， with the pathogenesis related to Qi and blood disharmony. The simultaneous disorders of Qi and blood are the key mechanism of MIRI， and thus the differentiation of Qi and blood syndromes is the prerequisite for the treatment. The theory of Qi and blood interacting in vessels is proposed by our team based on Qi being the commander of blood and blood being the mother of Qi as well as previous pharmacological studies. Specifically， Qi marshals blood by vessels， and the blood carries Qi by vessels. Accordingly， Qi and blood interact in the vessels. MIRI is accompanied by mitochondrial dysfunction， platelet function abnormality， and vascular endothelial damage， which are correlated with Qi deficiency， blood stasis， and vessel damage， respectively. Mitochondrial， platelet， and vascular endothelial structural and functional changes triggered by their interactions are one of the mechanisms by which Qi deficiency， blood stasis， and vessel damage lead to the occurrence and development of MIRI. By exploring the correlations between Qi and mitochondria， between blood and platelets， and between vessels and blood vessels， we can explain the modern scientific content of the theory of Qi and blood interacting in vessels in traditional Chinese medicine. According to the pathogenesis of Qi and blood disharmony in vessels， we discussed the pharmacological mechanisms of Qi-replenishing medicines， blood-activating medicines， and their combinations in the prevention and treatment of MIRI. On the basis of the research achievements in the prevention and treatment of MIRI by Qi-replenishing and blood-activating Chinese medicines based on the theory of Qi and blood interacting in vessels， we analyzed the effects of these medicines on Qi， blood， and vessels. According to the theory of Qi and blood， this article reveals the theoretical basis and scientific connotations of the prevention and treatment of cardiovascular diseases， with the aim of providing new ideas and references for the clinical application of traditional Chinese medicine in the prevention and treatment of cardiovascular diseases.