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

Objective To investigate the incidence and the types of gene mutations of α-thalassemia in the child-bearing pop-ulation of Conghua District,Guangzhou.Methods Blood samples from 24 083 people of childbearing age were screened by blood cell analysis and hemoglobin electrophoresis,α-globin gene variation was detected by GAP-PCR and PCR reverse dot blot in the positive cases,and 17 common β-globin gene mutations were detected by PCR reverse Dot blot.Results A total of 2 596 cases of α-thalassemia gene abnormality were detected by gene identification,and the abnormal rate was 10.78%.A sum of 170 cases(0.71%)had a compound mutation of α-β gene.There were 2 550 cases(98.23%)of deletion and 46 cases(1.77%)of non-deletion in the mutant genes.There were 14 types of gene mutation,including 5 types of HbH disease(with--SEA/-α3.7 primarily),4 mild types(with 68.61%of--SEA/αα genotype),and 5 quiescent types(the top two genotypes were-α3.7/αα and-α4.2/αα).A total of 23 types of αβ complex gene mutation were detected,and the top six types were--SEA/βCD41-42,-α3.7/βCD41-42,--SEA/β654,--SEA/-28,-α3.7/β654 and-α3.7/βCD17,which accounted for 75.27%of all the complex types.Conclusion The gene abnormality rate of α-thalassemia in Conghua District of Guangzhou City was high.The gene mutation type and constitu-ent ratio,which have their own characteristics,is a special region of α-thalassemia.

Objective To explore the application effect of"Internet + Medical treatment"model in patients with pulmonary rehabilitation after lung cancer surgery.Methods A total of 87 patients with lung cancer who received elective surgery in our hospital from June 2022 to January 2023 were selected and divided into control group(44 cases)and observation group(43 cases)according to random number table method.The control group received routine intervention,and the observation group received intervention based on"Internet + medical treatment"mode.The pulmonary function indexes[forced vital capacity(FVC),first second forced vital capacity(FEV1),respiratory flow(PEF)],cardiorespiratory endurance level[6min walking test(6MWT)],respiratory fatigue and quality of life(WHOQOL-BREF)of the two groups were compared.Results After 6 months of intervention,the scores of FVC,FEV1,PEF,6MWT distance and WHOQOL-BREE in the study group were higher than those in the control group,and the scores of respiratory fatigue were lower than those in the control group,with statistical significance(P<0.05).Conclusion"Internet + Medical treatment"intervention can improve lung function,increase cardiopulmonary endurance,reduce respiratory fatigue and improve quality of life in patients recovering from lung cancer surgery.

Objective To investigate the impact of the MAFB transcription factor on macrophage polarization phe-notype and function.Methods Analysis of correlation of MAFB expression with survival period of patients of breast cancer and lung adenocarcinoma was performed by RNA-seq data from the Kaplan-Meier database.Flow cytometry was applied to detect MAFB expression in primary human monocytes.Expression of MAFB mRNA or protein of hu-man acute monocytic leukaemia cell line(THP-1)was detected by RT-qPCR or Western blot.Detection of macro-phages phenotypes was performed by macrophages polarization in vitro,co-culture,RT-qPCR and flow cytometry.Phagocytosis assay was used to assess changes in phagocytic ability of macrophages.In vitro killing assay to evaluate changes in tumor-killing ability of macrophages.Results Compared to the MAFB low-expression group,patients with high MAFB expression in breast cancer and lung adenocarcinoma exhibited a decreased survival period(P＜0.05).Protein interaction networks showed a correlation between MAFB protein and macrophage polarization proteins.Compared to other cell type,MAFB was specifically highly expressed in human monocyte-macrophage cells.Phorbol myristate ester(PMA)induced an increased MAFB expression in THP-1 cells(P＜0.05).Knockdown of MAFB inhibited the expression of M2-related genes(P＜0.05).Co-culture with tumor cells and knockdown of MAFB suppressed expression of polarization gene(P＜0.05).Knockdown of MAFB did not affect the phagocytic ability P＞0.05)but enhanced tumor-killing ability of macrophages(P＜0.05).Conclusions This investigation proves potential function of MAFB transcription factor in regulation of macrophage M2 polarization and anti-tumor effect in solid tumors.

Objective:Based on the theory of demand analysis theory, to build teaching resources of human physiology course and explore its application effect.Methods:The three-year nursing junior college students of Batch 2018 were selected as the research objects. According to the principle of randomization, the students were divided into the observation group (97 cases) and control group (99 cases). Students in the observation group used integrated teaching resources from the perspective of demand analysis, while students in the control group used traditional curriculum resources. After the end of the course, the differences between the two groups of students' theoretical and practical assessment results and teaching experience were compared SPSS 20.0 was used for t test. Results:The results of theoretical [(73.67±8.76) vs. (70.48±8.36)] and practical skills [(84.41±4.83) vs. (83.63±4.27)] in the observation group were significantly different from those in the control group ( P<0.05). The overall scores of 9 teaching effect sub-items, such as learning awareness of inquiry, independent learning ability and critical thinking ability, were significantly higher than those of the control group ( P<0.01). Conclusion:The construction of human physiology teaching resources based on demand analysis theory can improve the effectiveness and pertinence of resource construction, and the effective use of resources in teaching can significantly improve the teaching effect, stimulate students' subjective initiative, cultivate their comprehensive ability and improve classroom efficiency.

Antimicrobial peptides (AMPs) are small molecule peptides that are widely found in living organisms with broad-spectrum antibacterial activity and immunomodulatory effect. Due to slower emergence of resistance, excellent clinical potential and wide range of application, AMP is a strong alternative to conventional antibiotics. AMP recognition is a significant direction in the field of AMP research. The high cost, low efficiency and long period shortcomings of the wet experiment methods prevent it from meeting the need for the large-scale AMP recognition. Therefore, computer-aided identification methods are important supplements to AMP recognition approaches, and one of the key issues is how to improve the accuracy. Protein sequences could be approximated as a language composed of amino acids. Consequently, rich features may be extracted using natural language processing (NLP) techniques. In this paper, we combine the pre-trained model BERT and the fine-tuned structure Text-CNN in the field of NLP to model protein languages, develop an open-source available antimicrobial peptide recognition tool and conduct a comparison with other five published tools. The experimental results show that the optimization of the two-phase training approach brings an overall improvement in accuracy, sensitivity, specificity, and Matthew correlation coefficient, offering a novel approach for further research on AMP recognition.
Anti-Bacterial Agents/chemistry* ; Amino Acid Sequence ; Antimicrobial Cationic Peptides/chemistry* ; Antimicrobial Peptides ; Natural Language Processing

AIM: To optimize an orthopedic non-muscle invasive bladder cancer (NMIBC) model in nude mouse by comparing four different ways of cellular transplantation, and to evaluate the efficacy of drug by bladder instillation, so as to provide a stable and efficient animal model for the treatment of bladder cancer. METHODS: After disruption of bladder mucosa by dilute acid-alkali or silver nitrate, T24 cells were instilled into the nude mouse bladder. T24 cells were injected directly into the bladder with mechanical injury of bladder mucosa. T24 cells were injected into the bladder wall. On the 14th day after making models, the nude mice were sacrificed. And the bladder mass and histopathological changes of tumor (including bladder) was observe to confirm the formation of orthopedic bladder cancer. The dynamic changes of orthopedic bladder cancer were observed after injecting T24 cells into the bladder wall. Gemcitabine was used to verify the applicability of the model of injecting T24 cells into the bladder wall in vivo. RESULTS: No tumor was found in the bladder after intravesical instillation of T24 cells with dilute acid-alkali or silver nitrate treatment. With mechanical injury of bladder mucosa, all nude mice had tumors after injection T24 cells. But the number of tumors varied and often occurred at multiple sites. The tumor was found in the bladder of all nude mice by injecting T24 cells into bladder wall, and there was only one tumor. The tumor showed slow linear growth within 15 days and rapid linear growth from day 18 to 31. In vivo efficacy evaluation, gemcitabine 150 mg/kg intravesical perfusion could significantly inhibit the growth of NMIBC in nude mice replicated by direct injection of T24 cells into the bladder wall, and the tumor inhibition rate was 97.1%. CONCLUSION: The orthotopic NMIBC model can not be established with the bladder mucosa injuried by dilute acid-alkali or silver nitrate treatment. The number and size of orthotopic bladder cancer are different by mechanical injury of bladder mucosa. Injection of T24 cells into the bladder wall of nude mouse can successfully establish the orthotopic NMIBC model, which can be used for the evaluation of NMIBC therapeutic drugs.

【Objective】 To analyze the polymorphisms of GYPA and GYPB mRNA spliceosomes associated with MNS blood group, and to explore the mechanism of subcellular localization of GPA and GPB protein isomerism encoded by various spliceosomes as well as the expression of MNS blood group antigen. 【Methods】 Ten blood samples of voluntary blood donors were randomly selected. The total mRNA of peripheral blood was extracted and reversed into cDNA. Nested PCR was used to amplify reading open frame of GYPA and GYPB gene, and sequencing was performed by Sanger. The base sequence obtained was compared with GYPA(NCBI: NM_002099) and GYPB(NCBI: Nm_002100.5). After the wild type and various splicing isomer of the open reading frame of GYPA and GYPB had been obtained, they were fused with the encoding gene of green fluorescent protein (GFP) by fusion PCR technology, then cloned and transfected into HEK293 cells for over expression. The subcellular localization of GPA-GFP and GPB-GFP fused fluorescent proteins was monitored by focusing laser scanning microscope. 【Results】 Exon-1 and Exon-2 were missing in GYPA mRNA of the 2 samples, and 2~26 amino acids were missing in the predicted GPA isomer, and the full length sequence of GYPB mRNA was complete. GYPA mRNA was intact in 6 samples, exon-2 was missing in GYPB mRNA, 13~45 amino acids were missing in the predicted GPB protein isomer, and other exon sequences were intact. One sample had intact GYPA mRNA, and 364~385 bases in exon-5 of GYPB mRNA were replaced by AG, indicating truncation of amino acid signal peptide. The GYP mRNA sequences of other samples were complete. The fluorescence signal of GP-GFP fusion protein showed that all GPA and GPB glycoprotein isomers, cloned according to various RNA splicing, could demonstrate the orientation distribution on the cell membrane surface, while some alternative splicing leaded to different degrees of protein dispersion in the cell, and affected the distribution speed and proportion of protein on the cell surface, which might be one of the reasons for the strength variation of MNS antigen. 【Conclusion】 The GYP mRNA spliceosome is obviously polymorphic, but the partial deletion of GYP mRNA fragment does not affect the localization and distribution of the protein isomers encoded by GYP mRNA on the cell surface, which can ensure the expression of MNS antigen characteristics.

Objective:To analyze the cases referred to a higher-level hospital from the Department of Neonatology in a primary hospital, and evaluate the efficiency of clinical works related to the referrals.Methods:Data of neonates admitted to the Department of Neonatology at Maternal and Child Health Hospital of Dabu County from January 2018 to December 2020 and referred to the superior hospital were retrospectively analyzed.Results:A total of 1 670 neonates were included and 128 neonates were referred.The median age of the neonates referred was 0.5 hours(0 hours, 25 days), the median gestational age was 38 + 3(29 + 1, 42 + 4) weeks, the median weight was 3 000(1 250, 4 800) g, and the transport distance was 78 km.Twenty-four cases were assessed as critical cases before the referral, 125 cases were improved and discharged after treatment in the superior hospital, and three cases died.The referral rate in 2018, 2019 and 2020 showed a downward trend year by year (10.3%, 7.6% and 4.0%, respectively), and the difference was statistically significant( χ2=14.362, P=0.001). The proportion of critical cases in referral cases increased year by year (9.4%, 23.9% and 38.9%, respectively), and the difference was statistically significant( χ2=9.289, P=0.010). The incidence of critical case was higher in those whose mothers didn′t have regular prenatal examination during pregnancy( χ2=5.129, P=0.032). Conclusion:The ability of neonatal treatment in primary hospitals has been improved.The neonates need to be referred and critical cases are not rare in primary hospitals.More attention should be paid to the safety and effectiveness of the regional transport network.Also, enhancing the health awareness of residents and improving the primary medical technics are important to maximize the life safety and optimal transition of newborns.

Objective:To explore the application of mixed BOPPPS (bridge-in, objective, pre-assessment, participatory-learning, post-assessment, summary) combined with case analysis teaching in nursing practice teaching in the department of endocrinology.Methods:Forty-five nurses of Batch 2017 who practiced in the Department of Endocrinology were selected as the control group, using traditional teaching; 45 practical nursing students of Batch 2018 were selected as the research group, using mixed BOPPPS combined with case analysis teaching. After the teaching, the teaching effect was evaluated by the theoretical and practical skill operation assessments, and the satisfaction with the teaching was investigated through the questionnaire. SPSS 22.0 was used for t test and chi-square test. Results:The scores of basic theory [(28.31±2.07) vs. (24.66±2.15)] and professional theory [(67.01±3.05) vs. (55.62±3.20)] in the theoretical assessment of practical nurses in the research group were higher than those in the control group, with significant differences ( P<0.05); the scores of each sub-item of the practical skills operation assessment were better than those in the control group, with significant differences ( P<0.05); in terms of teaching satisfaction, the research group was better than the control group in such 10 aspects as whether to improve the level of theoretical and practical skills operation, whether to improve the awareness of humanistic care and whether to improve the nurse-patient communication ability, etc., with significant differences ( P<0.05). Conclusion:The BOPPPS combined with case analysis teaching significantly improves the theoretical and practical skills and comprehensive literacy of the endocrinology practical nursing students, and the teaching satisfaction is significantly better than that of the traditional teaching, which is of promotion value.