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

Purpose/Significance To investigate the status quo and influencing factors of self-management level of elderly diabetic mellitus patients in Dongcheng District of Beijing,and to provide references for further development of community diabetes health educa-tion and health management.Method/Process The survey is conducted on 1 962 cases of elderly patients with type 2 diabetes mellitus who are randomly selected from community-level health service institutions in Dongcheng District of Beijing and are investigated by self-made questionnaires and the summary of diabetes self-care activities.Multiple stepwise regression and logistic regression are used to analyze the influencing factors of patients'self-management level.Result/Conclusion The self-management level of elderly diabetic mellitus patients is affected by many factors,among which the awareness of disease-related knowledge,the form of medical security and the use of health-related software to manage chronic diseases are the main influencing factors(P＜0.05).Local management depart-ments can take targeted health interventions to further improve the self-management ability of elderly diabetic mellitus patients.

"Toxicity Testing in the 21st Century—A Vision and Strategy"proposed by the National Research Council of US has brought innovative directives and objectives for toxicity evaluation and risk assessment,pushing forward the next generation of toxicity testing and risk assessment.In this initiative,the concept of adverse outcome pathways(AOPs)has emerged as a prominent methodology,capturing the attention of toxicologists and researchers due to its promising applications in recent years.The quantitative AOP(qAOP)is an extension of the adverse outcome pathway,which is built upon the foundational qualitative adverse outcome pathway model and leverages mathematical frame-works to depict dose-response and/or response-response relationships.This article reviews the princi-ples and advancement surrounding qAOP,introduceds two prevalent methodologies for constructing qAOP,Bayesian network models and regression models,and demonstrates diverse applications of qAOP.Actual cases are used to underscore the transformative role of qAOP in contemporary toxicology and risk assessment practices.

Objective:To investigate the effects of 30% ethanol elution fraction of Artemisia absinthium extract with macroporous resin (AAEM-30%) on the dendritic cell (DC) and immunity of mice. Methods:AAEM-30% was obtained from the alcoholic extracts of A. absinthium by AB-8 macroporous resin, and its polysaccharide, flavonoid, and terpenoid contents were determined. The expressions of AAEM-30% on DC surface molecular cluster of differentiation (CD) 40, CD80 and CD86 were detected in vitro by flow cytometry, and the expressions of DC cytokines IL-6 and tumor necrosis factor-α (TNF-α) were detected by enzyme linked immunosorbent assay (ELISA). The effect of AAEM-30% on the immune function of ICR mice was measured in vivo with different doses (50 and 100 mg/kg) and different administration methods (subcutaneous injection, intraperitoneal injection, and gavage). Results:The contents of polysaccharides, flavonoids, and terpenoids in AAEM-30% were 24.30%, 22.50% and 28.19%, respectively. AAEM-30% significantly enhanced the expression of CD40, and CD86 and the secretion of IL-6 and TNF-α (all P<0.001). Compared with the control group, no statistically significant differences were found in the body mass of mice compared with the three administration methods (all P>0.05). The thymus index in the 50 and 100 mg/kg AAEM-30% intraperitoneal injection groups and the spleen index in the 50 mg/kg AAEM-30% gavage group were increased (all P<0.05). CD19 + cells increased in the 100 mg/kg AAEM-30% intraperitoneal injection group ( P<0.01) and in the 50 mg/kg AAEM-30% gavage group ( P<0.05). The CD11b + and CD11c + counts increased in the 100 mg/kg AAEM-30% gavage group ( P<0.05). The number of CD4 + and CD8 + T lymphocytes was increased by both gavage and intraperitoneal administration (all P<0.05). Conclusions:AAEM-30% can promote the maturation of DC and enhanced the immunity of mice without obvious side effects.

Objective:To investigate the effect of silica gel column separation component of Artemisia asiatica (AEM-SC) on the maturation and immune function of mouse dendritic cells (DCs). Methods:Artemisia asiatica components were prepared by macroporous resin eluted with 70% ethanol, and then isolated by silica gel column to obtain AEM-SC. The contents of polysaccharides, flavonoids and triterpenes were quantified. Flow cytometry was used to detect the expression level of DCs surface molecules and antigen phagocytosis ability and to stimulate the proliferation of allogeneic T cells. ELISA method was used to detect the effect of DCs on cytokine secretion. Results:The contents of polysaccharides, flavonoids and triterpenes in AEM-SC were 10.12%, 5.7% and 3.62%, respectively. Functional tests showed that AEM-SC significantly reduced the expression levels of LPS-induced DCs surface molecules CD40, CD86 and MHC-II, reduced the expression levels of inflammatory cytokines IL-12p40, TNF-α and IL-6 (all P<0.05), improve the ability of phygocytosis ( P<0.01), and reduce the ability of DCs to stimulate the proliferation of CD4 +T and CD8 +T lymphocytes in the spleen of mice (all P<0.001). In the inflammatory mouse model experiment, AEM-SC significantly reduced the expression levels of DCs surface molecules CD40, CD86, CD80 (all P<0.001), and the expression levels of inflammatory cytokines TNF-α, IL-12p40 in serum (all P<0.01). Conclusions:AEM-SC can inhibit the maturation of DCs-induced LPS both in vitro and in vivo, indicating that AEM-SC has the immunosuppressive effect.

Objective To investigate the effects of Glycyrrhiza uralensis Fisch L. crude polysac-charides (GUCP) as an adjuvant on the immunity of mice and the immune responses induced by human pap-illoma virus ( HPV)-DNA vaccine. Methods ICR mice were injected with different concentrations of GUCP by different ways to detect the influences of GUCP on body weight, organ indexes and the numbers of immune cells in spleen. C57BL/ 6 mice were co-immunized with HPV-DNA vaccine and GUCP to detect the adjuvant efficacy on antigen-specific cellular and humoral immune responses. Results GUCP in all injected groups had no side effect on mouse body weight and liver, heart, lung and kidney indexes, but intraperitone-al injection of GUCP significantly increased spleen and thymus indexes and the numbers of B cells, CD4+ T cells, macrophages and dendritic cells in spleen. Subcutaneous injection of GUCP significantly increased the numbers of B cells and macrophages in spleen and intragastric administration significantly increased the num-bers of CD4+ and CD8+ T cells in spleen. Furthermore, GUCP as an adjuvant enhanced the antigen-specific CD4+ and CD8+ T cell responses and the levels of IgG, IgG1 and IgG2a induced by HPV-DNA vaccine at a certain degree. Conclusion GUCP enhanced the immunity of mice and the antigen-specific cellular and hu-moral immune responses induced by HPV-DNA vaccine. These results suggested that GUCP might be used as an adjuvant for DNA vaccine.

Objective:To evaluate the efficacy of individualized cognitive behavioral therapy for unprotected sex and sexual attitude of middle school students having unprotected sex.Methods:A target sample of 68 adolescents having unprotected sex was recruited from 4 secondary schools in Changsha,Hunan [the unprotected sex (US) score of Health-Risk Behavior Inventory for Chinese Adolescents (HBICA) ≥ 1].Subjects were randomized assigned to cognitive behavioral therapy group (CBT group) and control group.Each group had 34 subjects.The CBT group was giving one-on-one counseling for 6 weeks (50 to 60 minutes weekly).The control group didn't receive intervention by counselors.The US and Attitudes Toward Sexuality Scale (ATSS) were selected as criterion measurements.Outcome assessments were made at baseline and at 1-and 3-month follow-up.Results:The reduction rate of US scores showed that the response rate of therapy was over 80％.Mixed linear model analysis showed that there were significant group effect,time effect and group × time effect in scores of US and ATSS (Ps ＜ 0.05).Simple effect analysis indicated that the scores of US and ATSS of CBT group were significant lower than those of baseline from 1-month follow-up [(2.2 ± 2.9) vs.(4.7 ± 3.1),(3.2 ± 1.6) vs.(4.7 ± 3.1);(38.2 ± 4.9) vs.(40.9 ±5.1),(37.2 ±5.4) vs.(40.9 ±5.1),Ps ＜0.01],whereas the scores of those in the control group did not show any significant difference (Ps ＞0.05).At l-month and 3 month follow-up,moderate effect sizes were found for the CBT and control groups on all the outcome measures (Cohen's d =0.50-0.70).Conclusion:The individualized cognitive behavioral therapy could effectively reduce the level of unprotected sex and sexual attitude of adolescents having unprotected sex.

Objective The aim of this study was to determine the content of phthalate in disposable plastic tableware sold on Chengdu market,and to provide primary data for safety evaluation.Methods Sample selection was based on stratified sampling.Sixteen phthalate compounds were investigated in 60 disposable plastic tableware,divided into seven groups.The analysis was performed by gas chromatography-mass spectrometry (GC-MS).Results In this survey,diethyl phthalate,diisobutyl phthalate,dibutyl phthalate and diethylhexyl phthalate were detected,while the other 12 phthalate compound were not.The positive rates of the four detected phthalate were 6.7％ (4/60),10.0％ (6/60),46.7％ (28/60) and 28.3％ (17/60) respectively,and the highest concentrations were 10.3,6.4,7.2 and 65.6 mg/kg,respectively.Conclusion The observed level of detection rates and maximum concentrations were relatively high in this survey.In addition,some subgroups of PAEs that were not allowed to use in food contact materials were detected.Therefore,the migration in different food simulant would be analyzed in the next step for further health outcome assessment.

Objective: To explore the clinical significance of metastasis of lymph nodes between sternocleidomastoid and sternohyoid muscle (LNSS) in papillary thyroid cancer (PTC). Methods: A total of 175 patients with PTC who underwent thyroidectomy with LNSS dissection were retrospectively analyzed. Univariate and multivariate Logistic regression analyses were used to determine the independent risk factors for LNSS metastasis in PTC. Results: The rate of detectable LNSS was 70.9% (124/175) and metastasis rate was 7.4% (13/175). Of 13 cases with LNSS metastasis, 10 with the coexistence of cervical lymph node metastasis. Univariate Logistic regression analysis showed that multiple focal cancer, tumor located in the lower pole of thyroid, belt-shaped muscle invasion, lateral cervical lymph node metastasis, cN+ , the number of cervical lymph nodes with metastasis and the number of lymph nodes with metastasis in level Ⅳwere the risk factors for LNSS metastasis (P<0.05). Multivariate Logistic regression analysis suggested that tumor located in the lower pole of thyroid and the number of cervical lymph nodes with metastasis >6 were the independent risk factors for LNSS metastasis (P<0.05). Given the number of cervical lymph nodes with metastasis as a predictor for the LNSS metastasis, the sensitivity was 92.3%, the specificity was 66.7% and the accuracy rate was 68.6%. Conclusions LNSS metastasis is commom in PTC, with a metastasis rate of 7.4%. PTC in the lower pole of thyroid and the number of cervical lymph nodes with metastasis > 6 are independent risks for LNSS metastasis.

Objective To explore the effects of fasudil on inhibiting vascular smooth muscle cells (VSMCs) proliferation in vitro,increasing cell apoptosis,and inhibiting the Ras-MEK 1/2-ERK 1/2 pathway.Methods Healthy male SD rats (80～100 g) were selected.VSMCs were separated by the thoracoabdominal aortic vascular membrane dissection.Cultured VSMCs were randomly divided into 5 groups:serum-free group; serum group; serum + 1 μmol/L fasudil intervention group; serum + 10 μmol/L fasudil intervention group; serum + 100 μmol/L fasudil intervention group.The proliferation and migration of VSMCs were detected by MTT method and wound healing assay.Cell cycle and apoptosis of VSMCs were examined by flow cytometric analysis.The mRNA expressions of pro-apoptotic protein (Bax) and anti-apoptotic protein(Bcl-2) were determined by RT-PCR method,and the ratio of Bax/Bcl 2 was calculated.Western blot were performed to detect the protein expressions of Ras,MEK1/2,ERK1/2 and Akt in VSMCs.Results Fasudil inhibited rat VSMCs proliferation and migration,and blocked FBS-induced progression from the G0/G1 phase to S phase in a dose-dependent manner.Fasudil inhibited the early and late apoptosis in VSMCs,increased Bax mRNA expression and inhibited Bcl 2 mRNA expression.Fasudil significantly inhibited the protein expressions of FBS-stimulated intracellular Ras,phosphorylated MEK1/2,ERK1/2 in a dosedependent manner,but did not affect the protein expression of phosphorylated Akt.Conclusions Fasudil can attenuate VMSCs proliferation by blocking Ras-MEK1/2-ERK1/2 pathway and increasing cell apoptosis.