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 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.

【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 investigate the influence of all-trans-retinoic acid (ATRA) on the proliferation and invasion of osteosarcoma 143B cells and its possible regulatory mechanism.Methods:Different concentrations of ATRA were used to treat human osteosarcoma 143B cells, and the optimal concentration and treatment time those affected cell proliferation were selected. The MTS method, Transwell migration and invasion experiments were used to detect the changes in the proliferation, migration and invasion of 143B cells after ATRA treatment. The real-time quantitative polymerase chain reaction (qRT-PCR) and Western blot were used to detect the expression changes of miRNA-34a (miR-34a), E2F1 and Eag1 in osteosarcoma 143B cells after ATRA treatment. Then miR-34a was interfered and E2F1 was overexpressed, and the abilities of cell proliferation, invasion and migration abilities as well as the expression changes of miR-34a, E2F1 and Eag1 in 143B cells were detected.Results:The proliferation inhibition of 143B cells was most obvious when 143B cells were treated with 10 μmol/L ATRA for 72 h. The cell migration and invasion numbers when 143B cells were treated with 10 μmol/L ATRA for 72 h were lower than those in the negative control group [(73±3) cells vs. (182±5) cells, t = 21.46, P<0.01; (94±3) cells vs. (203±7) cells, t = 13.70, P<0.01]. 10 μmol/L ATRA could promote the expression of miR-34a in 143B cells and inhibit the expressions of Eag1 and E2F1 (all P<0.01). Compared with ATRA group, the ability of cell proliferation in ATRA+miR-34a interference group was restored after 72 h of treatment [cell survival rate (41.0±2.2)% vs. (25.0±3.6)%, t = 108.68, P<0.01]. Compared with ATRA group, the abilities of cell migration and invasion in ATRA+miR-34a interference group were restored [(122±14) cells vs. (64±10) cells, t = 21.06, P<0.01; (103±10) cells vs. (59±8) cells, t = 24.27, P<0.01), and the mRNA and protein expressions of Eag1 and E2F1 in cells were promoted (both P<0.01). Compared with ATRA group, the ability of cell proliferation in ATRA+E2F1 overexpression group was restored [cell survival rate (40.0±3.4)% vs. (24.0±3.1)%, t = 108.74, P<0.01]; the abilities of cell migration and invasion in ATRA+E2F1 overexpression group were restored [(78±12) cells vs. (29±8) cells, t = 13.52, P<0.01; (75±12) cells vs. (49±10) cells, t = 6.28, P<0.01], and the mRNA and protein expressions of Eag1 and E2F1 in cells were promoted (both P<0.01). Conclusion:ATRA inhibits the proliferation and invasion of osteosarcoma cells via regulating miR-34a-E2F1-Eag1 signaling pathway, and it may become one of the effective treatment drugs for osteosarcoma.

Objective: To Effects of n-hexane on learning and memory and the expressions of nerve growth factor (NGF) mRNA and nerve growth factor receptor (NGFR) mRNA of brain tissue in mice exposed to N-hexane. Methods: 40 Kunming mice were randomly divided into low-dose group, meddle-dose group, high-dose dose group and control group, with 10 mice in each group. All the groups were orally exposed to n-hexane in different doses: low-dose group with 43.5 mg/kg, middle-dose group with 86.5 mg/kg and high-dose group with 173.0 mg/kg, 1 time per day for 20 d. After the poisoning, the Y-arm test and the expressions of NGF mRNA and NGFR mRNA and the concentrations of NGF and NGFR in the brain tissues of each group were measured. Results: In the first Y-arm test, there existed a significant difference in correct reaction rate generally in all groups (P<0.05), and correct reaction rate in the middle-dose group and the high-dose group were low significantly compared with that in the control group(P<0.05). In the second Y-arm test, there existed a significant differences in total electric shock time and correct reaction rate generally in all groups (P<0.01), and the total electric shock time prolonged significantly and the correct response rate decreased significantly in 3 dose groups compared with those of the control group(P<0.05). The expression levels of NGF mRNA in brain tissues of low, meddle and high dose-groups were 0.81±0.66, 0.67±0.37 and 0.69±0.26, and the expression levels of NGFR mRNA were 1.22±0.42, 1.98±0.84 and 2.01±2.01, respectively. Compared with the control group, the expressions of NGF mRNA in the 3 dose groups decreased significantly (P<0.05), and the expression of NGFR mRNA in middle-and high-dose groups increased significantly (P<0.05). The concentrations of NGF in brain tissues of low,meddle and high dose-groups were 39.97±7.24 ng/L, 39.26±7.88 ng/L,31.70±8.21 ng/L,and the concentrations of NGFR were 17.37±6.82 ng/L,21.37±7.16 ng/L, 22.46±7.70 ng/L, respectively. Compared with the control group, the concentrations of NGF in high-dose groups decreased significantly(P<0.05), and the concentrations of NGFR in middle-and high-dose groups increased significantly (P<0.05). Conclusion N-hexane exposure can result in decrease of learning and memory in mice, which may be related to abnormal expression of NGF mRNA and NGFR mRNA in brain tissue.

Objectives To put forward the modified Letournel classification of acetabular fractures,and evaluate the guiding role of the modified Letournel classification in clinical work.Methods A retrospective study of 170 patients (178 sides) with acetabular fractures treated at Nanfang Hospital between January 2006 and August 2018 was performed.Among them 129 cases were males and 41 were females.The average age was 40.2±14.2 years (range,14-82 years).According to the Letournel classification,unclassifiable/atypical acetabular fractures were found out with plain Ⅹ-ray and plain CT scan and three-dimensional CT,and then their unclassifiable reasons were analyzed.Based on anatomical landmarks,clear boundaries of acetabular walls were defined on the 3D printed pelvic model.The structure of each column is a trihedron.The wall's fracture and column's fracture were distinguished using a theory that the wall's fracture involved two surfaces of column with interruption of continuity and column's fracture involved three.When column's fracture associated with wall's fracture,lowercase a,p and q were used representing comminuted areas of corresponding anterior and posterior wall,quadrilateral plate respectively.When column's fracture didn't associates with wall's fracture,it is marked with number 0.Finally,all fractures were classified according to the modified Letournel classification.The relationship between fracture type and surgical approach was analyzed.Results There was 51.7％ (92 sides) of acetabular fractures that couldn't be classified by the Letournel classification.Incomplete fracture lines (49 sides,53.3％),comminuted fractures (28 sides,30.4％),both of them (15 sides,16.3％) were their unclassifiable reasons.There were 8 types in the modified Letournel classification,including posterior wall fracture,posterior column fracture,anterior wall fracture,anterior column fracture,transverse fracture,T-shaped fracture,anterior column+posterior hemitransverse fracture,and both columns fracture.Posterior column+posterior wall fracture,transverse+posterior wall fracture had respectively become a form of posterior column fracture and of transverse fracture.According to the modified Letournel classification,the reclassification rate of 178 side fractures was 100％,which was significantly higher than 48.3％ (86/178) of the traditional Letournel classification,and the difference was statistically significant (x2=124.06,P＜ 0.001).100％ of posterior wall fracture and 80.00％ of posterior column fracture were treated by posterior approach.100％ of anterior column fracture and 73.68％ of both columns fracture were treated by anterior approach.Conclusion All acetabular fractures can be classified by the modified Letournel classification.The orientation of the comminuted walls reflects in part severity of the injury,better helps orthopaedic surgeons understand the morphology of acetabular fractures and select appropriate surgical approach.

Objective To investigate the influence of fat suppression technique on the parameters obtained with introvoxel incoherent motion (IVIM)for the normal hip.Methods 47 female volunteers were randomly selected to perform 1.5T MR examination with common sequence and IVIM sequence.The three different Fat suppression techniques:Fat sat,SPAIR and Water Excit were utilized respectively in T2WI sequence and IVIM sequence.The value of signal-to-noise ratio (SNR),contrast-to-noise ratio (CNR),coefficient of variation(CV), ADC,Dfast,Dslowand F values for three methods were calculated and compared quantitatively.The consistency evaluation of these methods were also conducted.Results (1)ADC value and Dslowvalue obtained with the Water Excit method were higher than those with Fat sat and SPAIR statistically(P<0.05),however there was no significant differences between the SNR,CNR,CV,Dfastand F values measured in three fat suppression methods(P>0.05).(2)The results of Fat sat and SPAIR techniques exhibited good consistency.Conclusion In Water Excit technique,overestimation of the ADC and Dslowvalues may be presented,while the IVIM measurement results are not influenced by the use of Fat sat or SPAIR method.

PURPOSE: MicroRNAs (miRs) were recently recognized to be important for immune cell differentiation and immune regulation. However, whether miRs were involved in allergen-specific immunotherapy (SIT) remains largely unknown. This study sought to examine changes in miR-146a and T regulatory cells in children with persistent allergic rhinitis (AR) after 3 months of subcutaneous immunotherapy (SCIT) and sublingual immunotherapy (SLIT). METHODS: Twenty-four HDM-sensitized children with persistent AR were enrolled and treated with SCIT (n=13) or SLIT (n=11) for 3 months. Relative miR-146a and Foxp3 mRNA expression, the TRAF6 protein level, and the ratio of post-treatment to baseline IL-10+CD4+ T cells between the SCIT and SLIT groups were examined in the peripheral blood mononuclear cells (PBMCs) of AR patients using quantitative reverse transcription polymerase chain reaction (qRT-PCR), flow cytometry, and Western blot analysis, respectively. Serum levels of IL-5 and IL-10 were determined using ELISA. RESULTS: After 3 months of SIT, both the TNSS and INSS scores were significantly decreased compared to the baseline value (P<0.01). The relative expression of miR-146a and Foxp3 mRNA was significantly increased after both SCIT and SLIT (P<0.01). The ratio of post-treatment to baseline IL-10+CD4+ T cells and the serum IL-10 level were significantly increased in both the SCIT and SLIT groups (P<0.01), whereas the TRAF6 protein level and serum IL-5 level were significantly decreased (P<0.01). No significant differences in these biomarkers were observed between the SCIT and SLIT groups. CONCLUSIONS: Our findings suggest that miR-146a and its related biomarkers may be comparably modulated after both SCIT and SLIT, highlighting miR-146a as a potential therapeutic target for the improved management of AR.
Biomarkers ; Blotting, Western ; Cell Differentiation ; Child* ; Enzyme-Linked Immunosorbent Assay ; Flow Cytometry ; Humans ; Immunotherapy* ; Interleukin-10 ; Interleukin-5 ; MicroRNAs ; Polymerase Chain Reaction ; Reverse Transcription ; Rhinitis* ; RNA, Messenger ; Sublingual Immunotherapy ; T-Lymphocytes ; TNF Receptor-Associated Factor 6

Aim To evaluate the effects of notoginsen-oside R1 on store-operated calcium entry ( SOCE ) in pulmonary arterial smooth muscle cells ( PASMCs ) of chronic hypoxia ( CH)-and monocrotaline ( MCT)-in-duced pulmonary hypertension ( PH) rats. Methods Mn2+ quenching of Fura-2 and measurement of intra-cellular free calcium concentration ( [ Ca2+] i ) using fluo-3 were examined in PASMCs of CH-exposed and MCT-treated rats. Results ①CH-exposed and MCT-treated rats exhibited profound PH when examined 3 weeks after hypoxia exposure or MCT injection, respec-tively. ②In the presence of 3 μmol·L-1 nifedipine, 10 μmol · L-1 notoginsenoside R1 significantly re-duced cyclopiazonic acid ( CPA )-induced the percent reduction in Fura-2 fluorescence measured 500 sec af-ter application of Mn2+, the maximal rate of Mn2+quenching, the amplitude of the Ca2+ influx transient and the resting [ Ca2+] i in PASMCs of CH-exposed and MCT-treated rats. Conclusion Notoginsenoside R1 inhibits SOCE and reduces resting [ Ca2+] i in PASMCs of CH-and MCT-induced PH rats.