Retrograde adeno-associated viruses (AAVs) are capable of infecting the axons of projection neurons and serve as a powerful tool for the anatomical and functional characterization of neural networks. However, few retrograde AAV capsids have been shown to offer access to cortical projection neurons across different species and enable the manipulation of neural function in non-human primates (NHPs). Here, we report the development of a novel retrograde AAV capsid, AAV-DJ8R, which efficiently labeled cortical projection neurons after local administration into the striatum of mice and macaques. In addition, intrastriatally injected AAV-DJ8R mediated opsin expression in the mouse motor cortex and induced robust behavioral alterations. Moreover, AAV-DJ8R markedly increased motor cortical neuron firing upon optogenetic light stimulation after viral delivery into the macaque putamen. These data demonstrate the usefulness of AAV-DJ8R as an efficient retrograde tracer for cortical projection neurons in rodents and NHPs and indicate its suitability for use in conducting functional interrogations.
Animals ; Haplorhini ; Axons ; Motor Neurons ; Interneurons ; Macaca ; Dependovirus/genetics* ; Genetic Vectors

BACKGROUND:Most of the silver coating materials prepared using active screen plasma technology in the past do not involve the nanotechnology field.The formed silver coating is in a"thin film"form,which is coated on the surface of the substrate,and the distribution of silver particles on the surface is uneven.Its long-term antibacterial ability is challenged. OBJECTIVE:To prepare nano silver coatings capable of being"buried"within stainless steel(SS)substrates using active screen plasma surface modification(ASPSM)and to observe antibacterial activity. METHODS:The nano-silver coating was prepared by ASPSM technique on stainless steel substrate.Three groups of coating samples were prepared by adjusting the bombardment time(1,2,and 4 hours),which were denoted as 1 h-Ag-ASPSM@SS,2 h-Ag-ASPSM@SS and 4 h-Ag-ASPSM@SS,respectively.The antibacterial activity of the coatings was analyzed by antibacterial ring test and Gram staining.The antibiotic coating samples of gentamicin combined with vancomycin were prepared by using stainless steel as substrate and were recorded as ACNs.Stainless steel,2 h-Ag-ASPSM@SS,and ACNs were inserted into Staphylococcus aureus or Pseudomonas aeruginosa suspension,respectively.The long-acting(84 days)antibacterial activity of the samples was analyzed by coating plate method.Bone marrow mesenchymal stem cells were co-cultured with stainless steel,2 h-Ag-ASPSM@SS,and ACNs,respectively.CCK-8 assay,dead/alive staining,and lactate dehydrogenase activity of cell supernatant were detected.Stainless steel,2 h-Ag-ASPSM@SS,and ACNs were taken after continuous exposure to Staphylococcus aureus suspension for 12 weeks.The amount of residual viable bacteria on the surface of the material was evaluated by spread plate method.Vancomycin drug sensitive disk method was used to evaluate the resistance of residual live bacteria on the surface of materials. RESULTS AND CONCLUSION:(1)With increasing bombardment time,the diameter of nano silver on the sample surface and the silver content in the coating gradually increased.Among them,the 2 h-Ag-ASPSM@SS exhibited the highest surface silver content while forming uniformly spherical nanoparticles.(2)Antibacterial ring test and Gram staining results demonstrated that compared with 1 h-Ag-ASPSM@SS and 4 h-Ag-ASPSM@SS,the 2 h-Ag-ASPSM@SS exhibited better inhibitory effect on Staphylococcus aureus and pseudomonas aeruginosa.After co-culturing with bacteria for 42 and 84 days,the number of viable bacteria on the spread plate method was significantly lower in the 2 h-Ag-ASPSM@SS group compared to the stainless steel and ACNs groups.After co-culturing with Staphylococcus aureus for 84 days and Pseudomonas aeruginosa for 42 days,the number of viable bacteria on the surface of the eluate from the ACNs group was higher than that of the stainless steel group.(3)CCK-8 assay,live/dead staining and lactate dehydrogenase activity of cell supernatant displayed that 2 h-Ag-ASPSM@SS did not have obvious cytotoxicity.ACNs showed obvious cytotoxicity.(4)After co-culture with Staphylococcus aureus for 12 weeks,the residual viable bacteria on the surface of 2 h-Ag-ASPSM@SS group was less than that of stainless steel group,and the residual viable bacteria on the surface of the ACNs group was more than that of stainless steel group.Compared with the stainless steel group,the sensitivity to vancomycin was significantly decreased in the ACNs group(P＜0.001),and there was no significant change in sensitivity to vancomycin in 2 h-Ag-ASPSM@SS group(P＞0.05).(5)The above results indicate that the silver nanoparticle coated stainless steel greatly improves the deposition efficiency of silver nanoparticles on the stainless steel surface and has long-lasting antibacterial properties and good cell compatibility.

Objective To explore the role of an artificial intelligence(AI)model based on real-time object detection network in fetal facial ultrasound examination.Methods With the normal fetal facial ultrasound standard plane(FFUSP)at 20-24 weeks of gestation as the research object,a FFUSP recognition model based on real-time object detection network was constructed.The recognition accuracy of the model for FFUSP and the anatomical structures were analyzed,and the clinical value was evaluated by analyzing its performance in identifying FFUSP in 119 cases of fetal ultrasound images.Results The overall precision,recall rate,mAP@.5 and mAP@.5:.95 of the AI model were 97.8%,98.5%,98.1%and 61.0%,respectively.The clinical validation showed that the AI model had a sensitivity,specificity,positive predictive value,negative predictive value and accuracy of 100.0%,98.5%,87.4%,100.0%and 98.7%for facial anatomy recognition,and the results were highly consistent with the classification of fetal ultrasound experts(k=0.925,P<0.001).The recognition accuracy of the model for 3 types of standard planes reached 100%;and the average speed of dynamic video detection was 33.93 frames per second.Conclusion The FFUSP recognition model based on real-time object detection network exhibits excellent performance,and it can be applied to real-time ultrasound diagnosis,teaching and intelligent quality evaluation.

[Objective] To evaluate the efficacy and safety of zero-pressure percutaneous nephrolithotomy (PCNL) combined with hands-free pneumatic lithotripsy in the one-stage treatment of calculous pyonephrosis in the non-acute infection phase. [Methods] Clinical data of 15 patients' treated during Feb.2022 and Dec.2023 were retrospectively analyzed.Double-sheath negative pressure PCNL was adopted, with zero-pressure gravity perfusion where the perfusion height was at the level of the patients' kidney, not exceeding 20 cm above the kidney.Pneumatic lithotripsy was performed without the need to hold the device with hand. [Results] The median stone diameter was 28 (range 22—40) mm, with a median stone density of 1028 (range 765—1305) Hu.All operations were successful.The median operation time was 55 (range 35—90) min.Postoperative low fever (Clavien grade Ⅰ) occurred in 1 case, and high fever (Clavien grade Ⅱ) occurred in 1 case, with no complications of Clavien grade Ⅲ or above.The initial stone-free rate was 73.3%, and the stone-free rate after one month was 93.3%. [Conclusion] For patients with calculous pyonephrosis in the non-acute infection phase, one-stage treatment using zero-pressure PCNL combined with hands-free pneumatic lithotripsy demonstrates good efficacy and safety.

[Objective] To summarize the clinical manifestations, pathological characteristics, treatment options and prognosis of the world's first case of prostate ductal adenocarcinoma (PDA) complicated with prostate mucinous adenocarcinoma (PMA). [Methods] The clinical and follow-up data of a patient with PDA and PMA treated in Zhongnan Hospital of Wuhan University were retrospectively analyzed, and relevant literature in PubMed and CNKI databases was retrieved. [Results] The patient sought medical attention due to dysuria, frequent urination, urinary urgency and urinary pain for more than half a year, and was admitted to hospital 3 times in total.The initial diagnosis upon the first admission was benign prostatic hyperplasia complicated with prostatic abscess.After 2 months, the patient was readmitted due to worsening symptoms, received transurethral bladder neck incision+ cystoscopy+ transurethral plasma resection of the prostate, and postoperative diagnosis confirmed PDA with local PMA.Three months after surgery, the patient had bleeding.After auxiliary examinations revealed extensive metastasis, he received hormonal therapy.After 9 months, the patient died due to multiple lung metastases. [Conclusion] Early diagnosis has a significant impact on the treatment and prognosis, but there have been no previous reports of PDA combined with PMA, so the lack of specific biomarkers in the early stage has led to missed diagnosis or misdiagnoses.There is no specific treatment for PDA with PMA. Radical prostatectomy was not satisfactory in the treatment of this case.

Objective To investigate the liver histopathological features of chronic hepatitis B(CHB)patients with normal alanine aminotransferase(ALT)and their correlation with serological markers.Methods Clinical data were collected from 137 patients with normal ALT who were treated in Wuxi Fifth People's Hospital from April 2018 to June 2021,and the differences in liver histopathology and serological markers were analyzed,as well as the correlation between liver histopathology and serological markers.The chi-square test was used for comparison of categorical data between groups,and the Kruskal-Wallis H test was used for comparison of data between multiple groups.A Spearman rank correlation test was performed,and logistic regression was used to perform the multivariate analysis.Results In the ALT≤20 U/L,20—29 U/L,and 30—40 U/L groups,the patients with significant inflammatory necrosis(≥G2)accounted for 57.4%,53.4%,and 75%,respectively,and the patients with significant fibrosis(≥S2)accounted for 63.8%,62.1%,and 75%,respectively.There was a significant difference in the degree of inflammatory necrosis between the patients with positive or negative HBeAg,the patients with different levels of serum HBV DNA,and the patients with different levels of serum HBV RNA(χ2=10.008,6.911,and 7.946,all P<0.05),and there was a significant difference in fibrosis stage between the patients with positive or negative HBeAg and the patients with different levels of serum HBV RNA(χ2=7.996 and 10.874,both P<0.05).The degree of liver inflammation and fibrosis stage were not significantly correlated with serum HBV DNA(rs=0.024,P=0.785;rs=0.039,P=0.652),while they were significantly correlated with serum HBV RNA(rs=0.222,P=0.009;rs=0.187,P=0.029).The multivariate analysis showed that in CHB patients,positive HBeAg was an independent risk factor for inflammatory necrosis(odds ratio[OR]=-0.302,95%confidence interval[CI]:-1.160 to 0.386,P=0.002)and fibrosis(OR=-0.387,95%CI:-1.160 to 0.386,P=0.011).Conclusion There are varying degrees of inflammatory necrosis and fibrosis in the liver of CHB patients with normal ALT,and positive HBeAg is independent risk factor for significant inflammatory necrosis and fibrosis in liver tissue of these patients.

This paper aims to focus on and implement strategies for preventing and resolving medical complaints and dis-putes at their emergence.The hospital promptly compiled a summary of the main problems and concerns reported from patients o-ver the past few years.It combined medical practices with the characteristics of admitted patients with severe diseases,complex treatment plans,and high risks.Since 2020,this hospital has been actively engaging with intractable high-risk patients and de-veloped a scheme that engaging clinical departments,medical management,and dispute management departments together to con-duct preoperative/pre-treatment communication.With a focus on patient management,this hospital upholds the concept of the pa-tient-centered humanistic philosophy in the process of doctor-patient communication to enhance the quality of communication.Consequently,the hospital has benefited from the communication scheme,achieving satisfactory effect in ensuring a solid medical safety and reducing the incidence of medical disputes.

Evaluating toxicity and decoding the underlying mechanisms of active compounds are crucial for drug development.In this study,we present an innovative,integrated approach that combines air flow-assisted desorption electrospray ionization mass spectrometry imaging(AFADESI-MSI),time-of-flight secondary ion mass spectrometry(ToF-SIMS),and spatial metabolomics to comprehensively investi-gate the nephrotoxicity and underlying mechanisms of nitidine chloride(NC),a promising anti-tumor drug candidate.Our quantitive AFADESI-MSI analysis unveiled the region specific of accumulation of NC in the kidney,particularly within the inner cortex(IC)region,following single and repeated dose of NC.High spatial resolution ToF-SIMS analysis further allowed us to precisely map the localization of NC within the renal tubule.Employing spatial metabolomics based on AFADESI-MSI,we identified over 70 discriminating endogenous metabolites associated with chronic NC exposure.These findings suggest the renal tubule as the primary target of NC toxicity and implicate renal transporters(organic cation transporters,multidrug and toxin extrusion,and organic cation transporter 2(OCT2)),metabolic en-zymes(protein arginine N-methyltransferase(PRMT)and nitric oxide synthase),mitochondria,oxidative stress,and inflammation in NC-induced nephrotoxicity.This study offers novel insights into NC-induced renal damage,representing a crucial step towards devising strategies to mitigate renal damage caused by this compound.

DEFENCATH?(taurolidine/heparin)is the first hemodialysis central venous catheter lock solution approved by the FDA.Taurolidine has broad-spectrum antibacterial activity against Gram-negative and Gram-positive bacterium and fungal species and anti-endotoxin and anti-exotoxin activities.Moreover,microbial resistance has not been observed,and the safety is good.Low-dose heparin is effective in anticoagulation and has a low risk of bleeding,and the two complement each other,clinical studies have reported that the catheter lock solution(CLS)effectively reduces the incidence of catheter-related blood stream infection(CRBSI),with good safety and significant clinical value.It will bring benefits to renal failure patients undergoing maintenance hemodialysis.

In the field of biomedicine,two-dimensional(2D)cell lines and animal models have played an im-portant role in the study of cell pathways and drug targets.However,due to species differences between humans and other animals,and the lack of hierarchy,cellular diversity,and cell-cell or cell-matrix interactions,2D cell lines could not ful-ly reflect what cells actually look like in the human body.Organoids are three-dimensional(3D)in vitro culture models de-rived from autologous tissue stem cells,which make up for the defects of 2D culture and can simulate the structure and function of real human organs to a certain extent,providing new ideas for disease diagnosis and treatment.Among them,lung organoids(LO)are a typical case studying the development process of human lung and the generation principle of lung diseases.Relevant studies have provided help for the treatment of pulmonary fibrosis,lung cancer,lung injury and other diseases.This paper aims to summarize and analyze the research progress of lung organoids in recent years,and fur-ther summarize the application of LO in the diagnosis and treatment of lung diseases.