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

Fucoidan is a kind of sulfated polysaccharide with various biological activities that mainly exists in the cell walls of brown algae. It is also found in marine invertebrates such as sea cucumbers and sea urchins. Fucoidan has received a lot of attention due to its tumor-killing and immune-boosting properties. Moreover, the combination of fucoidan with chemotherapeutic drugs not only improves antitumor efficacy but also reduces the side effects of these drugs. The function of fucoidan is closely correlated with its structure, molecular weight, degree of sulfation, monosaccharide component, algae source, and time of collection. In this review, the antitumor and immunomodulatory effects of fucoidan are reviewed from the aspects of promoting cell apoptosis, inducing cell cycle arrest, inhibiting angiogenesis and cell migration, and activating immune cells, to provide theoretical guidance for the development and clinical application of fucoidan.

Objective:To investigate the regulatory effects of nuclear factor-κB (NF-κB) on dendritic cell (DC) maturation and function through solute carrier family 1 member 2 (Slc1a2).Methods:Mouse bone marrow-derived DCs were transfected with Slc1a2-specific siRNA and an overexpression Slc1a2 eukaryotic expression vector. The real-time fluorescence quantitation (RT-PCR) and Western Blot methods were used to detect knockdown and overexpression efficiency. The expression of surface molecules (CD40, CD80) and major histocompatibility complex Ⅱ (MHCⅡ) of DCs was detected by flow cytometry. ELISA was used to detect the secretion of the cytokines interleukin (IL)-12, IL-6, and transforming growth factor-β (TGF-β). The effects of knockdown of Slc1a2 on DC maturation and function and the effects of overexpression of Slc1a2 on DC maturation and function were reflected by the above assay results. A mixed lymphocyte culture assay was used to investigate the effect of Slc1a2 on T cell proliferation, and an ELISA was used to detect the lavel of IL-17A. Changes in the relative fluorescence intensity of FITC in DCs were analyzed by flow cytometry to investigate the ability of Slc1a2 overexpression on antigen phagocytosis. Finally, DCs were pretreated with an NF-κB inhibitor, toluoylphenylalanine chloromethyl ketone (TPCK), and the effect of TPCK on the expression of Slc1a2 in DCs and DC maturation was examined.Results:Slc1a2 expression was found to be high in DC treated with lipopolysaccharides (LPS) ( P<0.001). The knockdown of Slc1a2 decreased DC maturation and ability to stimulate the proliferation of CD4 + T cells ( P<0.001) and inhibited IL-17 secretion ( P<0.01). Overexpression of Slc1a2 promoted DC maturation and ability to stimulate the proliferation of CD4 + T cells(all P<0.01) Pretreatment of DC with the NF-κB inhibitor TPCK inhibited the expression of Slc1a2 at mRNA and protein levels induced by LPS. Conclusions:NF-κB regulates Slc1a2 expression, which affects the maturation and function of DC.

Objective:To investigate the immunoregulatory effects of Glycyrrhiza uralensis ethanol extract(GUEE) on the maturation of dendritic cells (DCs) and the adjuvant effect of GUEE on OVA in na?ve BALB/c mice and an ovalbumin (OVA)-induced asthma mouse model. Methods:GUEE was prepared, and the effects of different concentrations of GUEE on the maturation of DCs and the secreted cytokines as well as the effects of GUEE on bacterial lipopolysaccharide (LPS)-induced DC maturation were examined in vitro. The effect of GUEE on the morphology of mouse bone marrow derived DCs was observed using microscopy. Molecular expression levels on the surface of DCs were detected using flow cytometry. The levels of interleukin-1β (IL-1β), IL-6, IL-12, and tumor necrosis factor-α (TNF-α) in the supernatant of DCs cultures were measured by enzyme-linked immunosorbent assay (ELISA). The maturation status of DCs was detected by flow cytometry by injecting different concentrations of GUEE into the paws of mice and isolating the draining lymph nodes 24 h later. The naive BALB/c mice were co-immunized with OVA, and the changes in regulatory T cells (Treg) were detected by flow cytometry. An OVA-protein-induced mouse asthma model was established to investigate whether GUEE as a tolerogenic adjuvant has an antigen-specific therapeutic effect on asthmatic mice. Pulmonary pathological changes were analyzed by hematoxylin-eosin staining (HE) and PAS staining. OVA-specific antibodies in serum and the frequencies of Tregs, CD4 + IFN-γ + and CD4 + IL-4 + T cells in the spleen were detected by ELISA and flow cytometry, respectively. Results:GUEE suppressed DCs maturation induced by LPS both in vitro and in vivo (all P<0.05), and reduced proinflammatory cytokine production, including IL-1β, IL-6, IL-12 and TNF-α in the absence or presence of LPS (all P<0.05). Moreover, co-immunization with OVA and GUEE increased the amount of Tregs in na?ve BALB/c mice ( P<0.05). In OVA-induced asthmatic mice, OVA and GUEE co-immunization and GUEE alone treatment substantially ameliorated the inflammation of lung tissues, decreased the levels of IgG 1 and the amount of CD4 + IL-4 + T cells, and increased the amount of Tregs (all P<0.05). Conclusions:GUEE alone or as the tolerogenic adjuvant can ameliorate allergic diseases through inhibition of DC maturation and type 2 helper T cell responses and induction of Tregs.

Objective To explore the clinical significance of monitoring drainage fluid parathyroid hormone (dPTH) for estimating the in situ reserves and function of the parathyroid by analyzing the change of serum calcium,serum parathyroid hormone(sPTH) and dPTH after thyroid surgery.Methods According to the operative method,the total of 144 patients with thyroid disease were divided into five groups:unilateral lobectomy,unilateral lobectomy plus isthmectomy with unilateral lymph node dissection,total thyroidectomy,total thyroidectomy with unilateral lymph node dissection,and total thyroidectomy with bilateral lymph node dissection group.The blood calcium,sPTH and dPTH level of patients were tested before operation and on the 1st,2nd,3rd and 4th day after operation.The depression of serum calcium,hypocalcemia and hypoparathyroidism were observed after operation.The serum calcium,serum PTH and dPTH level were summarized and analyzed statistically in order to evaluate the in situ reserves and postoperative function of the parathyroid.Results Among the 114 cases,the decline of serum calcium level mostly happened on the 2nd day after operation(70 cases,61.4％).There were 36 patients with hypocalcemia (31.58％) and 34 patients with hypoparathyroidism (29.82％).Serum calcium level increased gradually in all of the patients.Although sPTH level swung,it had a rising trend on the whole.The level of serum calcium and sPTH was positively correlated.The level of dPTH was discrete and decreased along with time.The decline level of dPTH among different groups had statistical difference.Conclusions It is a promising method to evaluate the in situ reserves and function of the parathyroid by monitoring the level and changes of dPTH after thyroid surgery,and it is of value for preventive calcium supplementation after thyroid surgery.

Objective To determine the correlation between phosphorylation extracellular signal-regulated kinase (p-Erk) and monopolar spindle 1 (Mps1) in colorectal cancer patients with the BRAF V600E mutation or not, and to explore the relationship between the expression of p-Erk and Mps1 with the clinicopathological features and prognosis of colorectal cancer patients. Methods Two hundred and eighty-eight paraffin-embedded tissue sections containing both the carcinoma and its adjacent non-neoplastic colorectal tissue were collected from January 2009 to June 2015 in the First Hospital of Shanxi Medical University. BRAF mutation was detected by Sanger sequencing. Kaplan-Meier survival analysis was used to analyze the correlation between BRAF V600E and prognosis. Immunohistochemistry was used to detect the expression level of p-Erk and Mps1 in colorectal cancer with wild type or BRAF V600E mutation. The correlation between p-ERK and Mps1 expression was analyzed by using linear regression analysis. Results The BRAF V600E mutation rate was 5.2 % in colorectal carcinomas. In addition, the poorly differentiated tumours and mucinous tumours had higher incidence of BRAF mutations than well differentiated tumours and non-mucinous tumours respectively [14.3 % (9/63) vs. 2.7 % (6/225),χ 2= 11.208,P = 0.001; 25.0 % (6/24) vs. 3.4 %(9/264),χ 2=16.630,P <0.001). The positive rate of p-Erk and Mps1 in colorectal carcinomas with BRAF V600E was significantly higher than that in colorectal carcinomas with BRAF WT (14/15 vs. 3/15, P <0.05; 15/15 vs. 3/15, P < 0.05]. It was found that the p-Erk expression correlated positively to the Mps1 expression (R2= 0.419,P < 0.001). The expressions of p-Erk protein in poorly differentiated adenocarcinoma was significantly higher and mucinous adenocarcinoma than those in high differentiated adenocarcinoma and non-mucinous adenocarcinoma (χ 2= 6.679, P = 0.01; χ 2= 5.735, P = 0.017), as well as in the group with lymph node metastasis than without lymph node metastasis (χ 2=5.436, P =0.02). Positive rate of Mps1 in poorly differentiated carcinoma was higher than that in well differentiated adenocarcinoma of colorectal carcinomas (χ 2=7.950, P =0.009). The Kaplan-Meier survival analysis indicated that patients with BRAF V600E had a worse survival rate than BRAF WT patients. Conclusions BRAF V600E may play an important role in specific pathological kinds of colorectal carcinomas, which is expected to be an independent prognostic factor. The expression of p-Erk is significantly correlated with Mps1 in colorectal carcinomas, suggesting that Mps1 may become a new potential target for targeted therapy.

Objective To examine the characteristics of affective intensity and cognitive emotion regulation in subjects with borderline personality disorder ( BPD) . Methods The BPD subscale of personal-ity disorder questionnaire( PDQ-4+) was used to assess the BPD symptoms,the short affective intensity scale (SAIS) and cognitive emotion regulation questionnaire (CERQ) were used to measure affect intensity and cognitive emotion regulation strategy,respectively. 765 subjects with BPD and 776 healthy controls were se-lected. The independent-samples t test was used to analyze the differences between BPD group and controls and logistic regression analysis was used to examine the related factors affecting the BPD. Results Com-pared with the control group(negative intensity:(3.08±0.66),negative coping dimension:(37.20±5.94), BPD group got higher scores in negative intensity((3.88±0.74), t=22.29, P<0.01,Cohen''s d>0.8)and negative coping dimension((44.77±6.36), t=24.16, P<0.01,Cohen''s d>0.8). The logistic regression anal-ysis showed that negative intensity(B=1.38,Exp(B)=3.97,95%CI for EXP(B):3.15~5.00, P<0.01) and negative cognitive regulations strategy(B=0.19,Exp(B)=1.21,95% CI:1.18-1.25, P<0.01) could affect the prevalence of BPD. Conclusion Subjects with BPD traits have more significant negative affective inten-sity and tend to use negative cognitive regulations strategy.