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 explore the effect and underlying mechanism of casein kinase 2 interacting protein-1 (CKIP-1) on hepatocyte apoptosis in nonalcoholic fatty liver disease (NAFLD).Methods:Experimental study. An NAFLD cell model was established by inducing human hepatoma cell line, HepG 2 cells, with oleic acid (OA). Flag-CKIP-1 expression vector and shRNA-CKIP-1 were transfected into HepG 2 cells. Flow cytometry was used to detect the effect of CKIP-1 on the activity and apoptosis of NAFLD hepatocytes. The levels of apoptosis-related proteins were detected by Western blot. CKIP-1 knockout mice in C57BL/6 back-ground were fed with either standard or high-fat diet for 8 weeks. Apoptosis-related signal proteins in NAFLD hepatocytes were detected by immunohistochemistry. Results:After CKIP-1 was transfected into HepG 2 cells, the degree of OA induced cell liposis was significantly reduced ( P<0.05). Annexin V-FITC/PI flow cytometry showed that CKIP-1 reduced the apoptosis of steatotic hepatocytes. Overexpression of CKIP-1 could significantly inhibit the expression of caspase-3 and caspase-9 and increase the expression of Bcl-2/Bax ( P<0.05). Knockdown of CKIP-1 could increase the expression of caspase-3 and caspase-9 ( P<0.05). CKIP-1 knockout could further increase the expression of caspase-3 and caspase-9 in NAFLD mice ( P<0.01, P<0.05), and further decrease the expression of Bcl-2/Bax ( P<0.05). Conclusion:CKIP-1 inhibited the apoptosis of steatotic hepatocytes by up-regulating the expression of apoptosis inhibitor gene, Bcl-2/Bax, and affecting the proteases, caspase-3 and caspase-9.

Fusobacterium nucleatum is an opportunistic pathogenic bacterium that can be enriched in colorectal cancer tissues, affecting multiple stages of colorectal cancer development. The two-component system plays an important role in the regulation and expression of genes related to pathogenic resistance and pathogenicity. In this paper, we focused on the CarRS two-component system of F. nucleatum, and the histidine kinase protein CarS was recombinantly expressed and characterized. Several online software such as SMART, CCTOP and AlphaFold2 were used to predict the secondary and tertiary structure of the CarS protein. The results showed that CarS is a membrane protein with two transmembrane helices and contains 9 α-helices and 12 β-folds. CarS protein is composed of two domains, one is the N-terminal transmembrane domain (amino acids 1-170), the other is the C-terminal intracellular domain. The latter is composed of a signal receiving domain (histidine kinases, adenylyl cyclases, methyl-accepting proteins, prokaryotic signaling proteins, HAMP), a phosphate receptor domain (histidine kinase domain, HisKA), and a histidine kinase catalytic domain (histidine kinase-like ATPase catalytic domain, HATPase_c). Since the full-length CarS protein could not be expressed in host cells, a fusion expression vector pET-28a(+)-MBP-TEV-CarS_cyto was constructed based on the characteristics of secondary and tertiary structures, and overexpressed in Escherichia coli BL21-Codonplus(DE3)RIL. CarS_cyto-MBP protein was purified by affinity chromatography, ion-exchange chromatography, and gel filtration chromatography with a final concentration of 20 mg/ml. CarS_cyto-MBP protein showed both protein kinase and phosphotransferase activities, and the MBP tag had no effect on the function of CarS_cyto protein. The above results provide a basis for in-depth analysis of the biological function of the CarRS two-component system in F. nucleatum.
Humans ; Histidine Kinase/metabolism* ; Fusobacterium nucleatum/metabolism* ; Automobiles ; Protein Kinases/genetics* ; Escherichia coli/metabolism* ; Colorectal Neoplasms

Implanted brain-computer interface (iBCI) is a system that establishes a direct communication channel between human brain and computer or an external devices by implanted neural electrode. Because of the good functional extensibility, iBCI devices as a platform technology have the potential to bring benefit to people with nervous system disease and progress rapidly from fundamental neuroscience discoveries to translational applications and market access. In this report, the industrialization process of implanted neural regulation medical devices is reviewed, and the translational pathway of iBCI in clinical application is proposed. However, the Food and Drug Administration (FDA) regulations and guidances for iBCI were expounded as a breakthrough medical device. Furthermore, several iBCI products in the process of applying for medical device registration certificate were briefly introduced and compared recently. Due to the complexity of iBCI in clinical application, the translational applications and industrialization of iBCI as a medical device need the closely cooperation between regulatory departments, companies, universities, institutes and hospitals in the future.
Humans ; Brain-Computer Interfaces ; Brain/physiology* ; Electrodes, Implanted

Objective To propose the method of dose distribution calculated by one-step optimization with 7 shells (Cao method) and compare with that by three-step optimization with 4 shells (Blanck method) and CyberKnife treatment plans for pancreatic cancer. Methods 20 cases of pancreatic cancer who underwent CyberKnife treatment were retrospectively analyzed,and CT was performed to localize and delineate the target area and endangering organs. Dosage was optimized and evaluated with Blanck method and Cao method. The planning target volume (PTV) conformity index (CI), new conformity index (nCI), homogeneity index (HI),gradient index (GI), coverage, dose-volume and doses to organs at risk were compared. Results Compared with Blanck method, CI (1.11 ± 0.05 vs 1.15 ± 0.05), nCI (1.20 ± 0.06 vs 1.23 ± 0.06), coverage [(92.48 ± 1.85)% vs (93.53 ± 2.15)%], volumes encompassed by 100% and 30% prescription dose line (36.46 ± 16.64 vs 38.19 ± 17.68; 286.19 ± 126.52 vs 320.93 ± 154.82) and monitor unit (56 369 ± 20 019 vs 57 814 ± 20 531) were significantly decreased,while GI was increased (3.22 ± 0.19 vs 3.11 ± 0.19), and all the differences were statistically significant (P<0.05). Additionally, Dmax of the intestine (21.17 ± 2.90 vs 20.63 ± 3.13), D10cc of the stomach (12.78 ± 2.57 vs 13.11 ± 2.43), D5ccof the duodenum (11.01 ± 3.45 vs 11.50 ± 3.25), D10ccof the duodenum (9.30 ± 3.31 vs 9.78 ± 3.07) and D0.35ccof the spinal cord (6.09 ± 0.98 vs 6.59 ± 0.92) were all significantly decreased (P<0.05). No significant differences were found on other parameters. Conclusions Better dose distributions are accessible by one-step optimization with 7 shells in CyberKnife treatment plans for pancreatic cancer.

Objective To explore the mechanisms of ligament of Marshall (LOM) initiat and sustain atrial fibrillation (AF).Methods The electrophysiologic properties of canine LOM were investigated using multipolar catheter mapping(normal canines,n =4,group A;AF canines,n =5,group B).The programmed stimulation were performed in the LOM,PV-left atrium(LA)junction and LA,respectively.Activations maps of LOM were analyzed from episodes of spontaneous onset of AF and initiation of induced AF by a single extrastimulus.The effectives refractory period of each part was compared and statistically analyzed among three parts in each group and between the two groups.LOM were cutted with surgical incision technology.The inducing rate of AF and the mapping rate of double potential and fragmented electrocardiogram were compared and statistically analyzed pro and post isolation of LOM.Results The incidence of abnormal potential of LOM in the two groups was significantly different(P ＜0.01),re-entry cycle(group A 25％ vs.B group 80％),tachycardia(group A 25％ vs.B 100％),double potential(group A 25％ vs.group B 80％),fragmentation potential(group A 25％ vs.group 80％).There was a significant difference in the rate of LOM tachycardia induction before and after LOM intervention in group B (P ＜ 0.05,before 100％ vs.after 20％).Conclusion There are two possible mechanisms of LOM involved in the occurrence and maintenance of AF:one is that LOM induces AF through spontaneous excitation,the other is that LOM participates in the reentry of left atrium and pulmonary vein in the form of bypass to induce and maintain AF.

Objective To study the clinical value of peripheral venous blood circulating tumor cell count in patients with non -small -cell lung cancer (NSCLC).Methods 28 patients with NSCLC were chosen as observa-tion group,at the same time,16 patients with benign lung tumor and 16 healthy persons were chosen as the control group 1,2.Before and after treatment,the peripheral venous blood circulating tumor cell counts were studied to under-stand its value in clinical.Results CK -FITC expression rate of adenocarcinoma patients was 95.8%,CD326APC expression rate was 94.5%,CK -FITC and CD326APC expression rate was 94.6%.CK -FITC expression rate of squamous carcinoma was 93.7%,CD326APC expression rate was 93.9%,CK -FITC and CD326APC expression rate was 94.1%,the differences between the two groups were not significant (χ2 =2.89,1.36,2.14,all P >0.05).The positive rate of peripheral venous blood circulating tumor cells in the observation group was significantly higher than the two control groups,and before treatment,the positive rate of peripheral venous blood circulating tumor cell was higher than after treatment.Conclusion Peripheral venous blood circulating tumor cell count can determine the dis-ease development process.

Objective To measure the effect of tetrandrineon (Tet) on inflammatory mediators and endogenous neural stem cell proliferation after spinal cord injury (SCI) in rats.Methods A total of 162 Wistar rats were separated into injury group,Tet group and sham operation group according to the random number table,with 54 rats per group.Allen' s method was used for induction of experimental SCI.Animals in Tet group were given Tet (22.5 mg/kg) through the tail vein at 30 min,24 h and 48 h postinjury.The same volume of normal saline was given to other two groups.Spinal cord tissue samples were taken from the rats after injury to measure levels of tumor necrosis factor (TNF)-α,interleukin (IL)-1 β and IL-10,and tissues were examined with HE staining and Nestin immunohistochemistry staining.Results Levels of TNF-α,IL-1 βand IL-10 in injury and Tet groups increased compared to these in sham operation group at 6 h,12 h,1 d,3 d,5 d and 1 week postinjury (P ＜ 0.05).At the same time point,level of IL-10 was higher in Tet group than in injury group,but inversely for TNF-α and IL-1 β (P ＜ 0.05).More Nestin-positive cells were present in injury and Tet groups than in sham operation group at 1 d,3 d,1 week,2 week,3 week and 4 week postinjury (P ＜ 0.05).Additionally,more Nestin-positive cells were found in Tet group than in injury group at 1 d,3 d,1 week,2 week and 3 week postinjury (P ＜ 0.05).Conclusion Tet is effective to relieve inflammatory reaction,increase neural stem cell number and promote neurological recovery after SCI.

Objective To investigate the influence and the efficacy for lipid level of rosuvastatin with atorv-astatin in the treatment of coronary heart disease.Methods 160 patients with coronary heart disease were randomly divided into the observation group and the control group,80 cases in each group.The patients of the observation group were given rosuvastatin 10mg every night,the control group were treated with the same dose of atorvastatin every night,the efficacy before and after treatment were compared after continuous 21d,and the lipid changed of the two groups were compared.Results There were statistically significant difference in the lipid level of the observation group before and after treatment(t =3.728,4.516,5.322,3.143,all P <0.05),and after treatment the TC,TG,LDL-C levels of the observation group were lower than those of the control group,the HDL -C level was higher than that of the control group,the difference was statistically significant(t =3.512,5.529,6.325,3.139,all P <0.05);There were statistically significant differences in hs -CRP,Hcy,IMT of the observation group before and after treatment [(14.35 ±1.56)μmol/L vs (22.45 ±1.82)μmol/L,(0.31 ±0.03)g/L vs (0.45 ±0.06)g/L,(1.64 ±0.11)mm vs (2.03 ±0.12)mm](t =3.728,3.231,3.452,all P <0.05),and the hs -CRP,Hcy,IMT of the observation group were lower than those of the control group[(14.35 ±1.56)μmol/L vs (16.56 ±1.26)μmol/L,(0.31 ±0.03)g/L vs (0.38 ±0.05)g/L,(1.64 ±0.11 )mm vs (1.81 ±0.12)mm](t =3.512,3.529,4.325,all P <0.05 ). Conclusion The treatment effect and lipid level of rosuvastatin was superior to atorvastatin.

Objective To discuss the effect of tetrandrine on endogenous neural stem cell proliferation and differentiation after spinal cord injury in rats. Methods 78 rats were randomly divided into 3 groups: control group(n=36), Tet-treated group(n=36), sham-operated group(n=6). Control group and Tet-treated group were adapted with Allen's combat modeling method. Rats in Tet group were injected Ted with a dosage 22.5 mg/kg in 30 minutes, 24 hours and 48 hours after ASCI, and the same dose of saline was injected into injured group as control .Samples were dissected from the spinal cord injury sites at 1 day, 3 days, 1 week, 2 weeks, 3 weeks, 4 weeks after ASCI, and tested by HE staining for morphology and by immunolfuorescence staining for the expression of BrdU and nestin. Results A little Nestin positive cells and BrdU positive cells were found in control group and Tet-treated group at 1 day after injury. A large number of positive cells were found in both groups at 1 week after injury and reached the peak which lasted for 2 weeks and then decreased gradually. The expression of Nestin positive cells and BrdU positive cells in control group and Tet-treated group were decreased significantly at 4 weeks after injury, but were still more than that in sham operation group. The number of Nestin positive cells and BrdU positive cells in Tet-treated group were more than that in control group at each time point after injury. The expression was higher in Tet-treated group than control group at 1 day, 3 days, 1 week, 2 weeks, 3 weeks after injury and had no difference at 4 weeks after injury. Conclusions Tetrandrine could increase the number of Nestin positive cells, BrdU positive cells and endogenous neural stem cells though improving the microenvironment, and it is beneficial for the recovery of spinal cord injury in rats.