Fluoro-ruby was injected into the posterior funiculus of the spinal cord in the cervical (C5-T2) and lumbar (L3-6) segments of adult guinea pigs. The spinal cord was cut into serial frozen sections. The Fluoro-rub...Fluoro-ruby was injected into the posterior funiculus of the spinal cord in the cervical (C5-T2) and lumbar (L3-6) segments of adult guinea pigs. The spinal cord was cut into serial frozen sections. The Fluoro-ruby labeling was clearly delineated from the surrounding structure. The labeling traversed the cervical, thoracic and lumbar segments, and was located on the ventral portion of the posterior funiculus on the injected side, proximal to the intermediate zone of the dorsal gray matter. The fluorescence area narrowed rostro-caudally. The spinal cord, spinal cord gray matter and corticospinal tract were reconstructed using 3D-DOCTOR 4.0 software, resulting in a robust three-dimensional profile. Using functionality provided by the reconstruction software, free multi-angle observation and sectioning could be conducted on the spinal cord and corticospinal tract. Our experimental findings indicate that the Fluoro-ruby retrograde fluorescent tracing technique can accurately display the anatomical location of corticospinal tract in the guinea pig and that three-dimensional reconstruction software can be used to provide a three-dimensional image of the corticospinal tract.展开更多
AIM:To assess the indications,findings,therapeutic procedures,safety,and complications of endoscopic retrograde cholangiopancreatography(ERCP) performed in Korean children.METHODS:The demographic characteristics,indic...AIM:To assess the indications,findings,therapeutic procedures,safety,and complications of endoscopic retrograde cholangiopancreatography(ERCP) performed in Korean children.METHODS:The demographic characteristics,indications for ERCP,findings,therapeutic procedures,and complications of 122 pediatric patients who underwent 245 ERCPs in the Asan Medical Center between June 1994 and March 2008 were investigated.RESULTS:The mean age of the 122 patients was 8.0 ± 4.2 years.Indications were biliary pathology in 78(64.0%),pancreatic pathology in 43(35.2%),and chronic abdominal pain in one.Biliary indications included choledochal cysts in 40,choledocholithiasis in 24,suspected sclerosing cholangitis in 8,trauma in 2,and other conditions in 4.Pancreatic indications includedacute pancreatitis in 7,acute recurrent pancreatitis in 11,chronic pancreatitis in 20,trauma in 3,and pancreatic mass in 2.Of the 245 ERCPs,success rate was 98.4% and 190(77.6%) were for therapeutic purposes,including endoscopic nasal drainage(51.8%),biliary sphincterotomy(38.0%),pancreatic sphincterotomy(23.3%),stent insertion(15.1%),stone extraction(18.8%),and balloon dilatation(11.0%).Complications were postERCP pancreatitis in 16(6.5%),ileus in 23(9.4%),hemorrhage in 2(0.8%),perforation in 2(0.8%),sepsis in 1(0.4%),and impacted basket in 1(0.4%).There were no procedure-related deaths,and most complications improved under supportive care.CONCLUSION:This study showed that there is a high incidence of choledochal cyst and diagnostic and therapeutic ERCP for the management of various biliary and pancreatic diseases was safe and effective in Korean children.展开更多
The purpose of this study was to investigate the effect of four fluorescent dyes, True Blue(TB), Fluoro-Gold(FG), Fluoro-Ruby(FR), and 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate(Di I...The purpose of this study was to investigate the effect of four fluorescent dyes, True Blue(TB), Fluoro-Gold(FG), Fluoro-Ruby(FR), and 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate(Di I), in retrograde tracing of rat spinal motor neurons. We transected the muscle branch of the rat femoral nerve and applied each tracer to the proximal stump in single labeling experiments, or combinations of tracers(FG-Di I and TB-Di I) in double labeling experiments. In the single labeling experiments, significantly fewer labeled motor neurons were observed after FR labeling than after TB, FG, or Di I, 3 days after tracer application. By 1 week, there were no significant differences in the number of labeled neurons between the four groups. In the double-labeling experiment, the number of double-labeled neurons in the FG-Di I group was not significantly different from that in the TB-Di I group 1 week after tracer application. Our findings indicate that TB, FG, and Di I have similar labeling efficacies in the retrograde labeling of spinal motor neurons in the rat femoral nerve when used alone. Furthermore, combinations of Di I and TB or FG are similarly effective. Therefore, of the dyes studied, TB, FG and Di I, and combinations of Di I with TB or FG, are the most suitable for retrograde labeling studies of motor neurons in the rat femoral nerve.展开更多
Analyzing the structure and function of the brain's neural network is critical for identifying the working principles of the brain and the mechanisms of brain diseases.Recombinant rabies viral vectors allow for th...Analyzing the structure and function of the brain's neural network is critical for identifying the working principles of the brain and the mechanisms of brain diseases.Recombinant rabies viral vectors allow for the retrograde labeling of projection neurons and cell type-specific trans-monosynaptic tracing,making these vectors powerful candidates for the dissection of synaptic inputs.Although several attenuated rabies viral vectors have been developed,their application in studies of functional networks is hindered by the long preparation cycle and low yield of these vectors.To overcome these limitations,we developed an improved production system for the rapid rescue and preparation of a high-titer CVS-N2c-ΔG virus.Our results showed that the new CVS-N2c-ΔG-based toolkit performed remarkably:(1)N2cG-coated CVS-N2c-ΔG allowed for efficient retrograde access to projection neurons that were unaddressed by rAAV9-Retro,and the efficiency was six times higher than that of rAAV9-Retro;(2)the trans-monosynaptic efficiency of oG-mediated CVS-N2c-ΔG was 2–3 times higher than that of oG-mediated SAD-B19-ΔG;(3)CVS-N2c-ΔG could delivery modified genes for neural activity monitoring,and the time window during which this was maintained was 3 weeks;and(4)CVS-N2c-ΔG could express sufficient recombinases for efficient transgene recombination.These findings demonstrate that new CVS-N2c-ΔG-based toolkit may serve as a versatile tool for structural and functional studies of neural circuits.展开更多
The new double projecting neurons were found in the cat spinal dorsal horn by the double retrograde fluorescent tracing technique.Fast Blue(FB)was injected into unilateral dorsal column nucleus(DCN)of adult cats anest...The new double projecting neurons were found in the cat spinal dorsal horn by the double retrograde fluorescent tracing technique.Fast Blue(FB)was injected into unilateral dorsal column nucleus(DCN)of adult cats anesthetized with pentobarbital.Nuclear Yellow(NY)was injected ipsilaterally into the lateral cervical nucleus(LCN)8-9 days later.After an additional 18-30 hrs.展开更多
In this study, biotinylated dextran amine (BDA) was microinjected into the left cortical motor area of the canine brain. Fluorescence microscopy results showed that a large amount of BDA-labeled pyramidal cells were...In this study, biotinylated dextran amine (BDA) was microinjected into the left cortical motor area of the canine brain. Fluorescence microscopy results showed that a large amount of BDA-labeled pyramidal cells were visible in the left cortical motor area after injection. In the left medulla oblongata, the BDA-labeled corticospinal tract was evenly distributed, with green fluorescence that had a clear boundary with the surrounding tissue. The BDA-positive corticospinal tract entered into the right lateral funiculus of the spinal cord and descended into the posterior part of the right lateral funiculus, close to the posterior horn, from cervical to sacral segments. There was a small amount of green fluorescence in the sacral segment. The distribution of BDA labeling in the canine central nervous system was consistent with the course of the corticospinal tract. Fluorescence labeling for BDA gradually diminished with time after injection. Our findings indicate that the BDA anterograde tracing technique can be used to visualize the localization and trajectory of the corticospinal tract in the canine central nervous system.展开更多
BACKGROUND Double-balloon endoscopic retrograde cholangiography (DB-ERC) is widely performed for biliary diseases after reconstruction in gastrointestinal surgery,but there are few reports on DB-ERC after hepatectomy ...BACKGROUND Double-balloon endoscopic retrograde cholangiography (DB-ERC) is widely performed for biliary diseases after reconstruction in gastrointestinal surgery,but there are few reports on DB-ERC after hepatectomy or living donor liver transplantation (LDLT).AIM To examine the success rates and safety of DB-ERC after hepatectomy or LDLT METHODS The study was performed retrospectively in 26 patients (45 procedures) who underwent hepatectomy or LDLT (liver operation:LO group) and 40 control patients (59 procedures) who underwent pancreatoduodenectomy (control group).The technical success (endoscope reaching the choledochojejunostomy site),diagnostic success (performance of cholangiography),therapeutic success(completed interventions) and overall success rates,insertion and procedure(completion of DB-ERC) time,and adverse events were compared between these groups.RESULTS There were no significant differences between LO and control groups in the technical [93.3%(42/45) vs 96.6%(57/59),P=0.439],diagnostic [83.3%(35/42) vs83.6%(46/55),P=0.968],therapeutic [97.0%(32/33) vs 97.7%(43/44),P=0.836],and overall [75.6%(34/45) vs 79.7%(47/59),P=0.617] success rates.The median insertion time (22 vs 14 min,P <0.001) and procedure time (43.5 vs 30 min,P=0.033) were significantly longer in the LO group.The incidence of adverse events showed no significant difference [11.1%(5/45) vs 6.8%(4/59),P=0.670].CONCLUSION DB-ERC after liver operation is safe and useful but longer time is required,so should be performed with particular care.展开更多
BACKGROUND Biliary drainage,either by the stent-in-stent(SIS)or side-by-side(SBS)technique,is often required when treating a malignant hilar biliary obstruction(MHBO).Both methods differ from each other and have disti...BACKGROUND Biliary drainage,either by the stent-in-stent(SIS)or side-by-side(SBS)technique,is often required when treating a malignant hilar biliary obstruction(MHBO).Both methods differ from each other and have distinct advantages.AIM To compare both techniques regarding their efficacy and safety in achieving drainage of MHBO.METHODS A comprehensive search of multiple electronic databases(MEDLINE,Embase,LILACS,BIREME,Cochrane)was conducted and grey literature from their inception until December 2020 with no restrictions regarding the year of publication or language,since there was at least an abstract in English.The included studies compared SIS and SBS techniques through endoscopic retrograde cholangiopancreatography.Outcomes analyzed included technical and clinical success,early and late adverse events(AEs),stent patency,reintervention,and procedure-related mortality.RESULTS Four cohort studies and one randomized controlled trial evaluating a total of 250 patients(127 in the SIS group and 123 in the SBS group)were included in this study.There were no statistically significant differences between the two groups concerning the evaluated outcomes,except for stent patency,which was higher in the SIS compared with the SBS technique[mean difference(d)=33.31;95%confidence interval:9.73 to 56.90,I2=45%,P=0.006].CONCLUSION The SIS method showed superior stent patency when compared to SBS for achieving bilateral drainage in MHBO.Both techniques are equivalent in terms of technical success,clinical success,rates of both early and late AEs,reintervention,and procedure-related mortality.展开更多
We investigated regeneration of the corticospinal tract and rearrangement of corticospinal nerves after spinal cord injury by biotinylated dextran amine (BDA) nerve tract tracing after neural stem cell transplantati...We investigated regeneration of the corticospinal tract and rearrangement of corticospinal nerves after spinal cord injury by biotinylated dextran amine (BDA) nerve tract tracing after neural stem cell transplantation. Neural stem cell transplantation increased motor function scores of rats at 3 weeks after spinal cord transection injury at the thoracic 10 segment. A proportion of BDA-labeled corticospinal tract regenerated through the spinal cord injury site at 12 weeks after transplantation. Electron microscopy revealed that the regenerated BDA-labeled nerve terminals formed new synaptic connections with neurons at the distal end of the injured site. These findings indicate that BDA nerve tract tracing effectively provides anatomic and morphological evidence of recovery after spinal cord injury.展开更多
Stroke can cause Wallerian degeneration in regions outside of the brain,particularly in the corticospinal tract.To investigate the fate of major glial cells and axons within affected areas of the corticospinal tract f...Stroke can cause Wallerian degeneration in regions outside of the brain,particularly in the corticospinal tract.To investigate the fate of major glial cells and axons within affected areas of the corticospinal tract following stroke,we induced photochemical infarction of the sensorimotor cortex leading to Wallerian degeneration along the full extent of the corticospinal tract.We first used a routine,sensitive marker of axonal injury,amyloid precursor protein,to examine Wallerian degeneration of the corticospinal tract.An antibody to amyloid precursor protein mapped exclusively to proximal axonal segments within the ischemic cortex,with no positive signal in distal parts of the corticospinal tract,at all time points.To improve visualization of Wallerian degeneration,we next utilized an orthograde virus that expresses green fluorescent protein to label the corticospinal tract and then quantitatively evaluated green fluorescent protein-expressing axons.Using this approach,we found that axonal degeneration began on day 3 post-stroke and was almost complete by 7 days after stroke.In addition,microglia mobilized and activated early,from day 7 after stroke,but did not maintain a phagocytic state over time.Meanwhile,astrocytes showed relatively delayed mobilization and a moderate response to Wallerian degeneration.Moreover,no anterograde degeneration of spinal anterior horn cells was observed in response to Wallerian degeneration of the corticospinal tract.In conclusion,our data provide evidence for dynamic,pathogenic spatiotemporal changes in major cellular components of the corticospinal tract during Wallerian degeneration.展开更多
Totally three articles focusing on magnetic resonance diffusion tensor imaging and diffusion tensor imaging evaluation of ischemic and hemorrhagic stroke, deep brain tumors and healthy corticospinal tract state are pu...Totally three articles focusing on magnetic resonance diffusion tensor imaging and diffusion tensor imaging evaluation of ischemic and hemorrhagic stroke, deep brain tumors and healthy corticospinal tract state are published in three issues. We hope that our readers find these papers useful to their research.展开更多
Objective: To study the correlation of serum trace element changes with inflammatory cytokines and oxidative stress indexes in patients with urinary tract infection (UTI). Methods: The patients who were diagnosed with...Objective: To study the correlation of serum trace element changes with inflammatory cytokines and oxidative stress indexes in patients with urinary tract infection (UTI). Methods: The patients who were diagnosed with lower urinary tract infection in our hospital between March 2013 and December 2017 were selected as the UTI group and the healthy volunteers who received physical examination were selected as the control group. Serum was collected to measure the contents of trace elements and inflammatory cytokines, and urine was collected to measure the contents of oxidative stress indexes. Results: The content of Zinc (Zn) in serum of the UTI group was significantly lower than that of the control group (P<0.05) while the contents of Copper (Cu), Selenium (Se), Calcium (Ca) and Magnesium (Mg) were not significantly different from those of the control group (P>0.05);the contents of C-reactive protein (CRP), amyloid protein A (SAA), procalcitonin (PCT), intercellular adhesion molecule-1 (ICAM-1), interleukin-2 (IL-2), interleukin-4 (IL-4), interleukin-5(IL-5), interleukin-17(IL-17) and transforming growth factor-β1 (TGF-β1) in serum as well as the contents of malondialdehyde (MDA) and advanced oxidation protein product (AOPP) in urine of the UTI group were significantly higher than those of the control group and negatively correlated with the content of Zn in serum (r=-0.514, -0.573, -0.475, -0.636, -0.612, -0.585,-0.492, -0.537, -0.611, -0.476 and -0.542, p=0.015, 0.011, 0.024, 0.007, 0.009, 0.010, 0.020, 0.014, 0.009, 0.024 and 0.013) while the contents of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and catalase (CAT) in urine were significantly lower than those of the control group and positively correlated with the content of Zn in serum (r=0.592, 0.657 and 0.610, p=0.010, 0.002 and 0.009). Conclusion: Serum trace element Zn significantly decreases in patients with urinary tract infection and it is closely related to the occurrence of disease as well as the abnormality of inflammation, immune response and oxidative stress in the course of the disease.展开更多
Rabies-viruses-based retrograde tracers can spread across multiple synapses in a retrograde direction in the nervous system of rodents and primates,making them powerful tools for determining the structure and function...Rabies-viruses-based retrograde tracers can spread across multiple synapses in a retrograde direction in the nervous system of rodents and primates,making them powerful tools for determining the structure and function of the complicated neural circuits of the brain.However,they have some limitations,such as posing high risks to human health and the inability to retrograde trans-synaptic label inputs from genetically-de¯ned starter neurons.Here,we established a new retrograde trans-multi-synaptic tracing method through brain-wide rabies virus glycoprotein(RVG)compensation,followed by glycoprotein-deleted rabies virus(RV-△G)infection in specific brain regions.Furthermore,in combination with the avian tumor virus receptor A(TVA)controlled by a cell-type-specific promoter,we found that EnvA-pseudotyped RV-△G can mediate e±cient retrograde trans-multi-synaptic transduction from cell-type-specific starter neurons.This study provides new alternative methods for neuroscience researchers to analyze the input neural networks of rodents and nonhuman primates.展开更多
Objective:We evaluated who would need further evaluations such as retrograde pyelography(RP)and/or ureteroscopy to diagnose upper urinary tract urothelial cancers(UUTUCs)when abnormal findings for the upper urinary tr...Objective:We evaluated who would need further evaluations such as retrograde pyelography(RP)and/or ureteroscopy to diagnose upper urinary tract urothelial cancers(UUTUCs)when abnormal findings for the upper urinary tract(UUT)were detected by enhanced computed tomography(CT).Methods:We retrospectively analyzed 125 patients who underwent enhanced CT for various reasons and had abnormal findings for the UUT.Patients whose tumors were suspected to be of extraureteral origin were excluded.All patients received RP and/or ureteroscopy to evaluate the UUTUCs.Results:The median age of the 125 patients was 70 years and gross hematuria(26.4%)was the most frequently observed symptoms.RP,ureteroscopy and both were performed for 121,59 and 55 patients,respectively.CT revealed tumor-like lesions in 58 patients and the other patients had non-tumor-like lesions.UUTUCs were found in 43(34.4%)of the 125 patients.All of them had tumor-like lesions on CT.In 58 patients who had tumor-like lesions on CT,univariate and multivariate analyses revealed that tumor diameter and tumor enhancement were significant predictive factors for UUTUCs.ROC curve analysis of enhanced CT to diagnose UUTUCs revealed that a tumor diameter of 18 mm was the best cutoff point.The sensitivity,specificity and accuracy were 90.0%,98.8% and 92.7% for RP and 95.5%,100% and 97.1%for ureteroscopy,respectively.Both of them had high sensitivity,specificity and accuracy.Conclusion:We should decide to evaluate the UUT according to the tumor diameter on enhanced CT.When we evaluate the UUT in patients with tumor diameters of less than 20 mm,ureteroscopy is recommended.展开更多
Efficient viral vectors for mapping and manipulating long-projection neuronal circuits are crucial in structural and functional studies of the brain. The SAD strain rabies virus with the glycoprotein gene deleted pseu...Efficient viral vectors for mapping and manipulating long-projection neuronal circuits are crucial in structural and functional studies of the brain. The SAD strain rabies virus with the glycoprotein gene deleted pseudotyped with the N2 C glycoprotein(SAD-RV(DG)-N2 C(G)) shows strong neuro-tropism in cell culture, but its in vivo efficiency for retrograde gene transduction and neuro-tropism have not been systematically characterized.We compared these features in different mouse brain regions for SAD-RV-N2 C(G) and two other widely-used retrograde tracers, SAD-RV(DG)-B19(G) and r AAV2-retro. We found that SAD-RV(DG)-N2 C(G) enhanced the infection efficiency of long-projecting neurons by^10 times but with very similar neuro-tropism, compared with SAD-RV(DG)-B19(G). On the other hand, SAD-RV(DG)-N2 C(G) had an infection efficiency comparable with r AAV2-retro, but a more restricted diffusion range, and broader tropism to different types and regions of longprojecting neuronal populations. These results demonstrate that SAD-RV(DG)-N2 C(G) can serve as an effective retrograde vector for studying neuronal circuits.展开更多
基金supported by a grant from the Priority Academic Program Development of Jiangsu Higher Education Institutions
文摘Fluoro-ruby was injected into the posterior funiculus of the spinal cord in the cervical (C5-T2) and lumbar (L3-6) segments of adult guinea pigs. The spinal cord was cut into serial frozen sections. The Fluoro-ruby labeling was clearly delineated from the surrounding structure. The labeling traversed the cervical, thoracic and lumbar segments, and was located on the ventral portion of the posterior funiculus on the injected side, proximal to the intermediate zone of the dorsal gray matter. The fluorescence area narrowed rostro-caudally. The spinal cord, spinal cord gray matter and corticospinal tract were reconstructed using 3D-DOCTOR 4.0 software, resulting in a robust three-dimensional profile. Using functionality provided by the reconstruction software, free multi-angle observation and sectioning could be conducted on the spinal cord and corticospinal tract. Our experimental findings indicate that the Fluoro-ruby retrograde fluorescent tracing technique can accurately display the anatomical location of corticospinal tract in the guinea pig and that three-dimensional reconstruction software can be used to provide a three-dimensional image of the corticospinal tract.
文摘AIM:To assess the indications,findings,therapeutic procedures,safety,and complications of endoscopic retrograde cholangiopancreatography(ERCP) performed in Korean children.METHODS:The demographic characteristics,indications for ERCP,findings,therapeutic procedures,and complications of 122 pediatric patients who underwent 245 ERCPs in the Asan Medical Center between June 1994 and March 2008 were investigated.RESULTS:The mean age of the 122 patients was 8.0 ± 4.2 years.Indications were biliary pathology in 78(64.0%),pancreatic pathology in 43(35.2%),and chronic abdominal pain in one.Biliary indications included choledochal cysts in 40,choledocholithiasis in 24,suspected sclerosing cholangitis in 8,trauma in 2,and other conditions in 4.Pancreatic indications includedacute pancreatitis in 7,acute recurrent pancreatitis in 11,chronic pancreatitis in 20,trauma in 3,and pancreatic mass in 2.Of the 245 ERCPs,success rate was 98.4% and 190(77.6%) were for therapeutic purposes,including endoscopic nasal drainage(51.8%),biliary sphincterotomy(38.0%),pancreatic sphincterotomy(23.3%),stent insertion(15.1%),stone extraction(18.8%),and balloon dilatation(11.0%).Complications were postERCP pancreatitis in 16(6.5%),ileus in 23(9.4%),hemorrhage in 2(0.8%),perforation in 2(0.8%),sepsis in 1(0.4%),and impacted basket in 1(0.4%).There were no procedure-related deaths,and most complications improved under supportive care.CONCLUSION:This study showed that there is a high incidence of choledochal cyst and diagnostic and therapeutic ERCP for the management of various biliary and pancreatic diseases was safe and effective in Korean children.
基金supported by a grants from the National Program on Key Basic Research Project(973 Program),No.2014CB542200the Innovative Research Team by the Ministry of Education,No.IRT1201+1 种基金the National Natural Science Foundation of China,No.31271284,31171150,81171146,30971526,31040043,31371210,81372044,31471144the Beijing Municipal Natural Science Foundation of China,No.7142164
文摘The purpose of this study was to investigate the effect of four fluorescent dyes, True Blue(TB), Fluoro-Gold(FG), Fluoro-Ruby(FR), and 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate(Di I), in retrograde tracing of rat spinal motor neurons. We transected the muscle branch of the rat femoral nerve and applied each tracer to the proximal stump in single labeling experiments, or combinations of tracers(FG-Di I and TB-Di I) in double labeling experiments. In the single labeling experiments, significantly fewer labeled motor neurons were observed after FR labeling than after TB, FG, or Di I, 3 days after tracer application. By 1 week, there were no significant differences in the number of labeled neurons between the four groups. In the double-labeling experiment, the number of double-labeled neurons in the FG-Di I group was not significantly different from that in the TB-Di I group 1 week after tracer application. Our findings indicate that TB, FG, and Di I have similar labeling efficacies in the retrograde labeling of spinal motor neurons in the rat femoral nerve when used alone. Furthermore, combinations of Di I and TB or FG are similarly effective. Therefore, of the dyes studied, TB, FG and Di I, and combinations of Di I with TB or FG, are the most suitable for retrograde labeling studies of motor neurons in the rat femoral nerve.
基金supported by the National Natural Science Foundation of China,Nos.32100899(to KZL),31830035(to FQX),31771156(to FQX),21921004(to FQX)the National Science and Technology Innovation 2030,No.2021ZD0201003(to FQX)+2 种基金the Key-Area Research and Development Program of Guangdong Province,No.2018B030331001(to FQX)the Strategic Priority Research Program of the Chinese Academy of Sciences,No.XDB32030200(to FQX)the Shenzhen Key Laboratory of Viral Vectors for Biomedicine,No.ZDSYS20200811142401005(to FQX)。
文摘Analyzing the structure and function of the brain's neural network is critical for identifying the working principles of the brain and the mechanisms of brain diseases.Recombinant rabies viral vectors allow for the retrograde labeling of projection neurons and cell type-specific trans-monosynaptic tracing,making these vectors powerful candidates for the dissection of synaptic inputs.Although several attenuated rabies viral vectors have been developed,their application in studies of functional networks is hindered by the long preparation cycle and low yield of these vectors.To overcome these limitations,we developed an improved production system for the rapid rescue and preparation of a high-titer CVS-N2c-ΔG virus.Our results showed that the new CVS-N2c-ΔG-based toolkit performed remarkably:(1)N2cG-coated CVS-N2c-ΔG allowed for efficient retrograde access to projection neurons that were unaddressed by rAAV9-Retro,and the efficiency was six times higher than that of rAAV9-Retro;(2)the trans-monosynaptic efficiency of oG-mediated CVS-N2c-ΔG was 2–3 times higher than that of oG-mediated SAD-B19-ΔG;(3)CVS-N2c-ΔG could delivery modified genes for neural activity monitoring,and the time window during which this was maintained was 3 weeks;and(4)CVS-N2c-ΔG could express sufficient recombinases for efficient transgene recombination.These findings demonstrate that new CVS-N2c-ΔG-based toolkit may serve as a versatile tool for structural and functional studies of neural circuits.
文摘The new double projecting neurons were found in the cat spinal dorsal horn by the double retrograde fluorescent tracing technique.Fast Blue(FB)was injected into unilateral dorsal column nucleus(DCN)of adult cats anesthetized with pentobarbital.Nuclear Yellow(NY)was injected ipsilaterally into the lateral cervical nucleus(LCN)8-9 days later.After an additional 18-30 hrs.
基金Fsupported by the Priority Academic Development Program of Jiangsu Higher Education Institutions
文摘In this study, biotinylated dextran amine (BDA) was microinjected into the left cortical motor area of the canine brain. Fluorescence microscopy results showed that a large amount of BDA-labeled pyramidal cells were visible in the left cortical motor area after injection. In the left medulla oblongata, the BDA-labeled corticospinal tract was evenly distributed, with green fluorescence that had a clear boundary with the surrounding tissue. The BDA-positive corticospinal tract entered into the right lateral funiculus of the spinal cord and descended into the posterior part of the right lateral funiculus, close to the posterior horn, from cervical to sacral segments. There was a small amount of green fluorescence in the sacral segment. The distribution of BDA labeling in the canine central nervous system was consistent with the course of the corticospinal tract. Fluorescence labeling for BDA gradually diminished with time after injection. Our findings indicate that the BDA anterograde tracing technique can be used to visualize the localization and trajectory of the corticospinal tract in the canine central nervous system.
文摘BACKGROUND Double-balloon endoscopic retrograde cholangiography (DB-ERC) is widely performed for biliary diseases after reconstruction in gastrointestinal surgery,but there are few reports on DB-ERC after hepatectomy or living donor liver transplantation (LDLT).AIM To examine the success rates and safety of DB-ERC after hepatectomy or LDLT METHODS The study was performed retrospectively in 26 patients (45 procedures) who underwent hepatectomy or LDLT (liver operation:LO group) and 40 control patients (59 procedures) who underwent pancreatoduodenectomy (control group).The technical success (endoscope reaching the choledochojejunostomy site),diagnostic success (performance of cholangiography),therapeutic success(completed interventions) and overall success rates,insertion and procedure(completion of DB-ERC) time,and adverse events were compared between these groups.RESULTS There were no significant differences between LO and control groups in the technical [93.3%(42/45) vs 96.6%(57/59),P=0.439],diagnostic [83.3%(35/42) vs83.6%(46/55),P=0.968],therapeutic [97.0%(32/33) vs 97.7%(43/44),P=0.836],and overall [75.6%(34/45) vs 79.7%(47/59),P=0.617] success rates.The median insertion time (22 vs 14 min,P <0.001) and procedure time (43.5 vs 30 min,P=0.033) were significantly longer in the LO group.The incidence of adverse events showed no significant difference [11.1%(5/45) vs 6.8%(4/59),P=0.670].CONCLUSION DB-ERC after liver operation is safe and useful but longer time is required,so should be performed with particular care.
文摘BACKGROUND Biliary drainage,either by the stent-in-stent(SIS)or side-by-side(SBS)technique,is often required when treating a malignant hilar biliary obstruction(MHBO).Both methods differ from each other and have distinct advantages.AIM To compare both techniques regarding their efficacy and safety in achieving drainage of MHBO.METHODS A comprehensive search of multiple electronic databases(MEDLINE,Embase,LILACS,BIREME,Cochrane)was conducted and grey literature from their inception until December 2020 with no restrictions regarding the year of publication or language,since there was at least an abstract in English.The included studies compared SIS and SBS techniques through endoscopic retrograde cholangiopancreatography.Outcomes analyzed included technical and clinical success,early and late adverse events(AEs),stent patency,reintervention,and procedure-related mortality.RESULTS Four cohort studies and one randomized controlled trial evaluating a total of 250 patients(127 in the SIS group and 123 in the SBS group)were included in this study.There were no statistically significant differences between the two groups concerning the evaluated outcomes,except for stent patency,which was higher in the SIS compared with the SBS technique[mean difference(d)=33.31;95%confidence interval:9.73 to 56.90,I2=45%,P=0.006].CONCLUSION The SIS method showed superior stent patency when compared to SBS for achieving bilateral drainage in MHBO.Both techniques are equivalent in terms of technical success,clinical success,rates of both early and late AEs,reintervention,and procedure-related mortality.
基金Hunan Province Science and Technology Department Plan,No.2009JT3051
文摘We investigated regeneration of the corticospinal tract and rearrangement of corticospinal nerves after spinal cord injury by biotinylated dextran amine (BDA) nerve tract tracing after neural stem cell transplantation. Neural stem cell transplantation increased motor function scores of rats at 3 weeks after spinal cord transection injury at the thoracic 10 segment. A proportion of BDA-labeled corticospinal tract regenerated through the spinal cord injury site at 12 weeks after transplantation. Electron microscopy revealed that the regenerated BDA-labeled nerve terminals formed new synaptic connections with neurons at the distal end of the injured site. These findings indicate that BDA nerve tract tracing effectively provides anatomic and morphological evidence of recovery after spinal cord injury.
基金supported by the National Natural Science Foundation of China,Nos.31 730030 (to XL),81941011 (to XL),31 771053 (to HD),82271403 (to XL),82272171 (to ZY),31971279 (to ZY)82201542 (to FH)+1 种基金the Natural Science Foundation of Beijing,No.7222004 (to HD)the Science and Technology Program of Beijing,No.Z181100001818007(to ZY)
文摘Stroke can cause Wallerian degeneration in regions outside of the brain,particularly in the corticospinal tract.To investigate the fate of major glial cells and axons within affected areas of the corticospinal tract following stroke,we induced photochemical infarction of the sensorimotor cortex leading to Wallerian degeneration along the full extent of the corticospinal tract.We first used a routine,sensitive marker of axonal injury,amyloid precursor protein,to examine Wallerian degeneration of the corticospinal tract.An antibody to amyloid precursor protein mapped exclusively to proximal axonal segments within the ischemic cortex,with no positive signal in distal parts of the corticospinal tract,at all time points.To improve visualization of Wallerian degeneration,we next utilized an orthograde virus that expresses green fluorescent protein to label the corticospinal tract and then quantitatively evaluated green fluorescent protein-expressing axons.Using this approach,we found that axonal degeneration began on day 3 post-stroke and was almost complete by 7 days after stroke.In addition,microglia mobilized and activated early,from day 7 after stroke,but did not maintain a phagocytic state over time.Meanwhile,astrocytes showed relatively delayed mobilization and a moderate response to Wallerian degeneration.Moreover,no anterograde degeneration of spinal anterior horn cells was observed in response to Wallerian degeneration of the corticospinal tract.In conclusion,our data provide evidence for dynamic,pathogenic spatiotemporal changes in major cellular components of the corticospinal tract during Wallerian degeneration.
文摘Totally three articles focusing on magnetic resonance diffusion tensor imaging and diffusion tensor imaging evaluation of ischemic and hemorrhagic stroke, deep brain tumors and healthy corticospinal tract state are published in three issues. We hope that our readers find these papers useful to their research.
文摘Objective: To study the correlation of serum trace element changes with inflammatory cytokines and oxidative stress indexes in patients with urinary tract infection (UTI). Methods: The patients who were diagnosed with lower urinary tract infection in our hospital between March 2013 and December 2017 were selected as the UTI group and the healthy volunteers who received physical examination were selected as the control group. Serum was collected to measure the contents of trace elements and inflammatory cytokines, and urine was collected to measure the contents of oxidative stress indexes. Results: The content of Zinc (Zn) in serum of the UTI group was significantly lower than that of the control group (P<0.05) while the contents of Copper (Cu), Selenium (Se), Calcium (Ca) and Magnesium (Mg) were not significantly different from those of the control group (P>0.05);the contents of C-reactive protein (CRP), amyloid protein A (SAA), procalcitonin (PCT), intercellular adhesion molecule-1 (ICAM-1), interleukin-2 (IL-2), interleukin-4 (IL-4), interleukin-5(IL-5), interleukin-17(IL-17) and transforming growth factor-β1 (TGF-β1) in serum as well as the contents of malondialdehyde (MDA) and advanced oxidation protein product (AOPP) in urine of the UTI group were significantly higher than those of the control group and negatively correlated with the content of Zn in serum (r=-0.514, -0.573, -0.475, -0.636, -0.612, -0.585,-0.492, -0.537, -0.611, -0.476 and -0.542, p=0.015, 0.011, 0.024, 0.007, 0.009, 0.010, 0.020, 0.014, 0.009, 0.024 and 0.013) while the contents of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and catalase (CAT) in urine were significantly lower than those of the control group and positively correlated with the content of Zn in serum (r=0.592, 0.657 and 0.610, p=0.010, 0.002 and 0.009). Conclusion: Serum trace element Zn significantly decreases in patients with urinary tract infection and it is closely related to the occurrence of disease as well as the abnormality of inflammation, immune response and oxidative stress in the course of the disease.
基金study was supported by the STI2030-Major Projects(Grant No.2021ZD0201003)the National Natural Science Foundation of China(Grant Nos.31830035,31771156,21921004,and 32100899)+3 种基金the Strategic Priority Research Program of the Chinese Academy of Sciences(Grant No.XDB32030200)the Shenzhen Key Laboratory of Viral Vectors for Biomedicine(Grant No.ZDSYS20200811142401005)the Key Laboratory of Quality Control Technology for Virus-Based Ther-apeutics,Guangdong Provincial Medical Products Administration(Grant No.2022ZDZ13)Open Project Program of Wuhan National Laboratory for Optoelectronics(Grant No.2019WNLOKF022).
文摘Rabies-viruses-based retrograde tracers can spread across multiple synapses in a retrograde direction in the nervous system of rodents and primates,making them powerful tools for determining the structure and function of the complicated neural circuits of the brain.However,they have some limitations,such as posing high risks to human health and the inability to retrograde trans-synaptic label inputs from genetically-de¯ned starter neurons.Here,we established a new retrograde trans-multi-synaptic tracing method through brain-wide rabies virus glycoprotein(RVG)compensation,followed by glycoprotein-deleted rabies virus(RV-△G)infection in specific brain regions.Furthermore,in combination with the avian tumor virus receptor A(TVA)controlled by a cell-type-specific promoter,we found that EnvA-pseudotyped RV-△G can mediate e±cient retrograde trans-multi-synaptic transduction from cell-type-specific starter neurons.This study provides new alternative methods for neuroscience researchers to analyze the input neural networks of rodents and nonhuman primates.
文摘Objective:We evaluated who would need further evaluations such as retrograde pyelography(RP)and/or ureteroscopy to diagnose upper urinary tract urothelial cancers(UUTUCs)when abnormal findings for the upper urinary tract(UUT)were detected by enhanced computed tomography(CT).Methods:We retrospectively analyzed 125 patients who underwent enhanced CT for various reasons and had abnormal findings for the UUT.Patients whose tumors were suspected to be of extraureteral origin were excluded.All patients received RP and/or ureteroscopy to evaluate the UUTUCs.Results:The median age of the 125 patients was 70 years and gross hematuria(26.4%)was the most frequently observed symptoms.RP,ureteroscopy and both were performed for 121,59 and 55 patients,respectively.CT revealed tumor-like lesions in 58 patients and the other patients had non-tumor-like lesions.UUTUCs were found in 43(34.4%)of the 125 patients.All of them had tumor-like lesions on CT.In 58 patients who had tumor-like lesions on CT,univariate and multivariate analyses revealed that tumor diameter and tumor enhancement were significant predictive factors for UUTUCs.ROC curve analysis of enhanced CT to diagnose UUTUCs revealed that a tumor diameter of 18 mm was the best cutoff point.The sensitivity,specificity and accuracy were 90.0%,98.8% and 92.7% for RP and 95.5%,100% and 97.1%for ureteroscopy,respectively.Both of them had high sensitivity,specificity and accuracy.Conclusion:We should decide to evaluate the UUT according to the tumor diameter on enhanced CT.When we evaluate the UUT in patients with tumor diameters of less than 20 mm,ureteroscopy is recommended.
基金the National Basic Research Program(973 Program)of China(2015CB755601)the Strategic Priority Research Program of the Chinese Academy of Sciences(XDB32030200)+1 种基金the National Natural Science Foundation of China(31771156,81661148053,91632303,31800885,31500868,31671120 and 91732304)the China Postdoctoral Science Foundation(2019M653118 and 2018M632946).
文摘Efficient viral vectors for mapping and manipulating long-projection neuronal circuits are crucial in structural and functional studies of the brain. The SAD strain rabies virus with the glycoprotein gene deleted pseudotyped with the N2 C glycoprotein(SAD-RV(DG)-N2 C(G)) shows strong neuro-tropism in cell culture, but its in vivo efficiency for retrograde gene transduction and neuro-tropism have not been systematically characterized.We compared these features in different mouse brain regions for SAD-RV-N2 C(G) and two other widely-used retrograde tracers, SAD-RV(DG)-B19(G) and r AAV2-retro. We found that SAD-RV(DG)-N2 C(G) enhanced the infection efficiency of long-projecting neurons by^10 times but with very similar neuro-tropism, compared with SAD-RV(DG)-B19(G). On the other hand, SAD-RV(DG)-N2 C(G) had an infection efficiency comparable with r AAV2-retro, but a more restricted diffusion range, and broader tropism to different types and regions of longprojecting neuronal populations. These results demonstrate that SAD-RV(DG)-N2 C(G) can serve as an effective retrograde vector for studying neuronal circuits.