Purpose–As an important part of the management of railway passenger transport,the rationality and effectiveness of the clearing method of railway passenger transport are directly related to the operating efficiency a...Purpose–As an important part of the management of railway passenger transport,the rationality and effectiveness of the clearing method of railway passenger transport are directly related to the operating efficiency and service quality of railway passenger transport enterprises.This paper aims to comprehensively and deeply discuss the evolution and development process of China’s railway passenger transport clearing method,analyze its characteristics and influences in each stage,identify the main factors affecting its evolution and development and then put forward thoughts on improving the future development of the clearing method.Design/methodology/approach–Through a detailed review of the railway passenger transport clearing methods from the planned economy period to the reform and opening up period and into the new century,the basis,mode,subject and object of clearing in different development stages are systematically compared.Findings–It comprehensively reveals the evolution of the clearing method,sorted out the characteristics and changes of the clearing method at each stage and the adaptability to the development of railway passenger transport at that time.The characteristics of the development of clearing measures for railway passenger transport in different stages and their far-reaching influence on railway passenger transport business are deeply analyzed.Originality/value–This paper summarized the factors influencing the development of China’s railway passenger transportation clearing approach evolution,including the simplified rules of clearing,enhanced the market adaptability,establishing and perfecting the incentive mechanism,strengthening the construction of informatization,etc.This paper puts forward the ways to improve the railway passenger transportation clearing future development thinking.展开更多
Vascular-targeted photodynamic therapy(V-PDT)is an effective treatment for port wine stains(PWS).However,repeated treatment is usually needed to achieve optimal treatment outcomes,possibly due to the limited treatment...Vascular-targeted photodynamic therapy(V-PDT)is an effective treatment for port wine stains(PWS).However,repeated treatment is usually needed to achieve optimal treatment outcomes,possibly due to the limited treatment light penetration depth in the PWS lesion.The optical clearing technique can increase light penetration in depth by reducing light scattering.This study aimed to investigate the V-PDT in combination with an optical clearing agent(OCA)for the therapeutic enhancement of V-PDT in the rodent skinfold window chamber model.Vascular responses were closely monitored with laser speckle contrast imaging(LSCI),optical coherence tomography angiography,and stereo microscope before,during,and after the treatment.We further quantitatively demonstrated the effects of V-PDT in combination with OCA on the blood flow and blood vessel size of skin microvasculature.The combination of OCA and V-PDT resulted in significant vascular damage,including vasoconstriction and the reduction of blood flow.Our results indicate the promising potential of OCA for enhancing V-PDT for treating vascular-related diseases,including PWS.展开更多
Citrus yellow vein clearing virus(CYVCV)is a new citrus virus that has become an important factor restricting the development of China’s citrus industry,and the CYVCV coat protein(CP)is associated with viral pathogen...Citrus yellow vein clearing virus(CYVCV)is a new citrus virus that has become an important factor restricting the development of China’s citrus industry,and the CYVCV coat protein(CP)is associated with viral pathogenicity.In this study,the Eureka lemon zinc finger protein(ZFP)ClDOF3.4 was shown to interact with CYVCV CP in vivo and in vitro.Transient expression of ClDOF3.4 in Eureka lemon induced the expression of salicylic acid(SA)-related and hypersensitive response marker genes,and triggered a reactive oxygen species burst,ion leakage necrosis,and the accumulation of free SA.Furthermore,the CYVCV titer in ClDOF3.4 transgenic Eureka lemon plants was approximately 69.4%that in control plants 6 mon after inoculation,with only mild leaf chlorotic spots observed in those transgenic plants.Taken together,the results indicate that ClDOF3.4 not only interacts with CP but also induces an immune response in Eureka lemon by inducing the SA pathways.This is the first report that ZFP is involved in the immune response of a citrus viral disease,which provides a basis for further study of the molecular mechanism of CYVCV infection.展开更多
Three-dimensional(3D)cell cultures have contributed to a variety of biological research fields by filling the gap between monolayers and animal models.The modern optical sectioning microscopic methods make it possible...Three-dimensional(3D)cell cultures have contributed to a variety of biological research fields by filling the gap between monolayers and animal models.The modern optical sectioning microscopic methods make it possible to probe the complexity of 3D cell cultures but are limited by the inherent opaqueness.While tissue optical clearing methods have emerged as powerful tools for investigating whole-mount tissues in 3D,they often have limitations,such as being too harsh for fragile 3D cell cultures,requiring complex handling protocols,or inducing tissue deformation with shrinkage or expansion.To address this issue,we proposed a modified optical clearing method for 3D cell cultures,called MACS-W,which is simple,highly efficient,and morphology-preserving.In our evaluation of MACS-W,we found that it exhibits excellent clearing capability in just 10 min,with minimal deformation,and helps drug evaluation on tumor spheroids.In summary,MACS-W is a fast,minimally-deformative and fluorescence compatible clearing method that has the potential to be widely used in the studies of 3D cell cultures.展开更多
Organisms are composed of a vast number of highly specialized cells with characteristic physiological functions determined by their structure.Therefore,understanding physiological and pathological mechanisms requires ...Organisms are composed of a vast number of highly specialized cells with characteristic physiological functions determined by their structure.Therefore,understanding physiological and pathological mechanisms requires obtaining 3-dimensional(3D)structural information at the cellular level across tissues,organs,and even the whole body.However,the scattering and absorption of tissue limits the penetration depth of light and thus extremely reduces the imaging performance in deep tissue.It is well known that 3D imaging outperforms 2D imaging in various scenarios for 3D reconstruction of tissue structures.This includes tracing convoluted 3D structures like vasculature,neurons,and prostate glands,as well as elucidating intricate distributions of cells or lesions.展开更多
[Objective] The research aimed to study a new whole clearing and preparation technology.[Method] After young seeds of Stephania sp.preserved in the alcohol solution,as well as the fresh pistil and nucellus of Sorghum ...[Objective] The research aimed to study a new whole clearing and preparation technology.[Method] After young seeds of Stephania sp.preserved in the alcohol solution,as well as the fresh pistil and nucellus of Sorghum bicolor were separated,they were soaked and cleared in the solution of 84 disinfectant.And the plant materials could be mounted by using three methods.[Result] The globular embryos and heart-shaped embryos at different stages of Stephania sp.as well as the pistil,nucellus,embryo sac,hypostase o...展开更多
MATLAB software and optimal complete subgraph algorithm were used to extract and reveal the microsatellite distribution features in the complete genomes of the tobacco vein clearing virus （NC-003 378.1） from the NCB...MATLAB software and optimal complete subgraph algorithm were used to extract and reveal the microsatellite distribution features in the complete genomes of the tobacco vein clearing virus （NC-003 378.1） from the NCBI database.The results showed that the repetitions number and their location of the N-base group has been extracted and displayed.The largest repetitions of N-base group in the complete genomes of the tobacco vein clearing virus was decreased as the exponential function with the increasing of N.The method used in this study could be applied to the extraction and revealing of the microsatellite distribution features in the complete genomes of other viruses,thereby provided a basis for the research of the structure and the law of function,inheritance and variation by the using of the microsatellite distribution features.展开更多
For the first time,the changes in autofluorescence spectra of ex vivo rat skin have been experimentally investigated using the combination of fluorescence spectroscopy and optical immersion clearing.The glucose,glycer...For the first time,the changes in autofluorescence spectra of ex vivo rat skin have been experimentally investigated using the combination of fluorescence spectroscopy and optical immersion clearing.The glucose,glycerol and propylene glycol solutions were used as clearing agents.The optical clearing was performed from the dermal side of skin imitating the in vivo injection of clearing agent under the dermal layers.In this contribution,the common properties of autofluorescence variation during optical immersion clearing were determined.The tendency of autofluorescence signal to decrease with reduction of scattering in tissue was noticed and discussed in detail.However,the differences in the shape of spectral curves under application of different clearing agents showed that optical clearing affects the autofluorescence properties of tissue differently depending on the type of clearing liquid.The results obtained are useful for the understanding of tissue optical clearing mechanisms and for improving techniques such as fluorescence spectroscopy.展开更多
Tissue optical clearing by use of optical clearing agents(OCAs)has been proven to have potential to reduce the highly scattering effect of biological tissues in optical techniques.However,the difference in tissue samp...Tissue optical clearing by use of optical clearing agents(OCAs)has been proven to have potential to reduce the highly scattering effect of biological tissues in optical techniques.However,the difference in tissue samples could lead to unreliable results,making it difficult to quantitatively control the dose of OCAs during the course of tissue optical clearing.In this work,in order to study the effects of optical clearing,we customized tissue-like phantoms with optical properties of some biological tissue.Diffuse reflectance and total transmittance of tissue-like phantoms with different OCAs(DMSO or glycerol)and porcine skin tissues were measured.Then optical property parameters were calculated by inverse adding-doubling(IAD)algorithm.Results showed that OCAs could lead to a reduction in scattering of tissue-like phantoms as it did to porcine skin tissue in vitro.Furthermore,a series of relational expressions could be fit to quantitatively describe the relationship between the doses of OCAs and the reduction of scattering effects.Therefore,proper tissue-like phantom could facilitate optical clearing to be used in quantitative control of tissue optical properties,and further promote the application potential of optical clearing to light-based noninvasive diagnostic and therapeutic techniques.展开更多
Citrus yellow vein clearing virus （CYVCV） is considered as the causal agent of Citrus yellow vein clearing disease and belongs to the genus Mandarivirus in the family Alphaflexiviridae. Capsid protein （CP） of CYVC...Citrus yellow vein clearing virus （CYVCV） is considered as the causal agent of Citrus yellow vein clearing disease and belongs to the genus Mandarivirus in the family Alphaflexiviridae. Capsid protein （CP） of CYVCV Chongqing isolate （CYVCV- CQ） was produced using a prokaryotic expression system and used as the immunogen for monoclonal antibody （MAb） production. Four highly specific and sensitive murine MAbs and one polyclonal antibody were prepared in this study. Titers of the four MAbs in ascites fluids ranged from 10-6 to 10-7 as determined by indirect enzyme-linked immunosorbent assay （ELISA）. Three serological assays, including dot enzyme-linked immunosorbent assay （dot-ELISA）, tissue blot-ELISA, and double-antibody sandwich （DAS）-ELISA, were developed for quick and reliable detections of CYVCV in citrus samples. The developed dot-ELISA and DAS-ELISA methods could detect CYVCV in the infected citrus leaf crude extracts diluted at 1：2 560 and 1：10 240 （w/v, g mL^-1）, respectively. The detection result of 125 citrus leaf samples collected from citrus groves in Yunnan Province and Chongqing Municipality of China showed that approximately 36% samples were positive for CYVCV. This virus was, however, not＇detected in any sample collected from Zhejiang or Jiangxi Province, China.展开更多
OBJECTIVE: To assess the efficacy and safety in 105 patients with seasonal Influenza in Beijing, the mixture prepared with Chinese medicines follows the treatment regimen of releasing exterior cold andclearing interio...OBJECTIVE: To assess the efficacy and safety in 105 patients with seasonal Influenza in Beijing, the mixture prepared with Chinese medicines follows the treatment regimen of releasing exterior cold andclearing interior heat.METHODS: Total 330 patients with seasonal influenza were enrolled and randomly and averagely divided into the Chinese herbal medicine, the western medicine and the Chinese patent medicine group. They were treated with Chinese medicine Oseltamivir Phosphate Capsules and the Scattering Wind and Resolving Toxins Capsules. The main efficacy indicators were the antifebrile onset time and recovery time of body temperature. The efficacy and safety of the mixture was scientifically evaluated. Comparisons of several variables were analyzed.RESULTS: Median antifebrile onset time of the Chinese herbal medicine group was significantly shorter than the western medicine group(P < 0.05) and the Chinese patent medicine group(P < 0.05). The median antifebrile recovery time of the Chinese herbal medicine group was significantly shorter than the Chinese patent medicine group(P <0.05). The groups evaluated by TCM symptom pattern effect, both the Chinese herbal medicine group and Western Medicine group were better than the Chinese patent medicine group(P <0.05). The disappearance rate of main symptoms and some minor symptom patterns of the Chinese herbal medicine group were higher than the other 2 groups.CONCLUSION: The mixture of releasing exterior cold and clearing interior heat could significantly shorten the fever time with safety.展开更多
Morphological analyses are key outcome assessments for nerve regeneration studies but are historically limited to tissue sections.Novel optical tissue clearing techniques enabling three-dimensional imaging of entire o...Morphological analyses are key outcome assessments for nerve regeneration studies but are historically limited to tissue sections.Novel optical tissue clearing techniques enabling three-dimensional imaging of entire organs at a subcellular resolution have revolutionized morphological studies of the brain.To extend their applicability to experimental nerve repair studies we adapted these techniques to nerves and their motor and sensory targets in rats.The solvent-based protocols rendered harvested peripheral nerves and their target organs transparent within 24 hours while preserving tissue architecture and fluorescence.The optical clearing was compatible with conventional laboratory techniques,including retrograde labeling studies,and computational image segmentation,providing fast and precise cell quantitation.Further,optically cleared organs enabled three-dimensional morphometry at an unprecedented scale including dermatome-wide innervation studies,tracing of intramuscular nerve branches or mapping of neurovascular networks.Given their wide-ranging applicability,rapid processing times,and low costs,tissue clearing techniques are likely to be a key technology for next-generation nerve repair studies.All procedures were approved by the Hospital for Sick Children’s Laboratory Animal Services Committee(49871/9)on November 9,2019.展开更多
Objective： To explore the clinical effect of therapy of clearing hallow viscera in treating critical patients with gastro-enteric function disorder （GEFD）. Methods： Retrospective analysis was carried out on 96 cri...Objective： To explore the clinical effect of therapy of clearing hallow viscera in treating critical patients with gastro-enteric function disorder （GEFD）. Methods： Retrospective analysis was carried out on 96 critical patients. They were 48 patients in the treated group treated with Dachengqi Decoction （ 大承气汤） and 48 patients in the control group treated with Western medicine for promoting gastric dynamic force. The recovery rate, recovery time of gastro-enteric function, incidence rate and fatality rate of multiple organ dysfunction syndrome （MODS）, as well as the level of plasma endotoxin （ET） before and after treatment between the two groups were compared. Results： Comparison between the two groups in gastro-enteric function recovery rate （81.3% vs 45.8 %）, functional disorder sustaining time in patients who got recovered （1.2± 0.3 daysvs4.0±1.1 days）, incidence rate （29. 17% vs 52.08%） and fatality rate （28.57% vs 56.00%） of MODS all showed significant difference （ P〈0.05 or P〈0.01 ）. The plasma level of ET after treatment in the treated group was significantly lower than that in the control group （P〈0.05）. Conclusion： Therapy of clearing hallow viscera has a good effect in treating critical patients with gastro-enteric function disorder, and could reduce the incidence and fatality of MODS.展开更多
Successful establishment of reconnection between retinal ganglion cells and retinorecipient regions in the brain is critical to optic nerve regeneration.However,morphological assessments of retinorecipient regions are...Successful establishment of reconnection between retinal ganglion cells and retinorecipient regions in the brain is critical to optic nerve regeneration.However,morphological assessments of retinorecipient regions are limited by the opacity of brain tissue.In this study,we used an innovative tissue cleaning technique combined with retrograde trans-synaptic viral tracing to observe changes in retinorecipient regions connected to retinal ganglion cells in mice after optic nerve injury.Specifically,we performed light-sheet imaging of whole brain tissue after a clearing process.We found that pseudorabies virus 724(PRV724)mostly infected retinal ganglion cells,and that we could use it to retrogradely trace the retinorecipient regions in whole tissue-cleared brains.Unexpectedly,PRV724-traced neurons were more widely distributed compared with data from previous studies.We found that optic nerve injury could selectively modify projections from retinal ganglion cells in the hypothalamic paraventricular nucleus,intergeniculate leaflet,ventral lateral geniculate nucleus,central amygdala,basolateral amygdala,Edinger-Westphal nucleus,and oculomotor nucleus,but not the superior vestibular nucleus,red nucleus,locus coeruleus,gigantocellular reticular nucleus,or facial nerve nucleus.Our findings demonstrate that the tissue clearing technique,combined with retrograde trans-synaptic viral tracing,can be used to objectively and comprehensively evaluate changes in mouse retinorecipient regions that receive projections from retinal ganglion cells after optic nerve injury.Thus,our approach may be useful for future estimations of optic nerve injury and regeneration.展开更多
Citrus yellow vein clearing virus(CYVCV)is the causal agent of yellow vein clearing disease,a significant and devastating disease of most citrus species including lemon and sour orange.CYVCV,a single-strand positive-s...Citrus yellow vein clearing virus(CYVCV)is the causal agent of yellow vein clearing disease,a significant and devastating disease of most citrus species including lemon and sour orange.CYVCV,a single-strand positive-sense RNA virus containing six ORFs(Open Reading Frames),represents a new species in the genus Mandarivirus of the Alphaflexiviridae family.The virus can cause particularly serious damage,resulting in reduced tree vigor,lower yields,and decreased marketability of fruit production,and it has been found in India,Turkey,Pakistan,China and Iran.Here we described the geographical distribution of the virus,its transmission mode by grafting,mechanical inoculation,and insects,as well as currently available techniques for detection.In addition,we also discussed practical measures aimed at controlling the disease and provided theoretical guidance to prevent the acquisition and spread of the disease that is a significant step toward ensuring the health of the citrus industry.展开更多
To identity the potential pathogen associated with the yellow vein clearing symptom on lemon trees, the profiles of virus-de- rived small interfering RNAs from citrus samples were obtained and analyzed by deep sequenc...To identity the potential pathogen associated with the yellow vein clearing symptom on lemon trees, the profiles of virus-de- rived small interfering RNAs from citrus samples were obtained and analyzed by deep sequencing method in this study. Twenty-seven contigs almost cover the full length genome of Citrus yellow vein clearing virus （CYVCV） isolate YN were obtained using the small RNA deep sequencing technology. Analysis showed that this isolate CQ shared the highest nucleotide identity with isolate Y1 （JX040635） and YN （KP313242）, both of which belong to the genus Mandarivirus in the family Alphaflexiviridae. Mapping analysis of viral-derived siRNA （vsiRNA） profiles revealed an uneven distribution pattern of their generations along both positive and negative genome strands, and 22- and 21-nt vsiRNAs ranked the majority. BLAST against viroids and other viral databases confirmed that this sample was single-infected only by CYVCV, which indicated that CYVCV was the exact causal agent for the yellow clearing symptom occurring on lemon. This is the first CYVCV isolate detected in Chongqing and the second in China. This result could provide a molecular basis for the investigation of citrus viral diseases to protect citrus health in this region.展开更多
Confocal Raman microspectroscopy(CRM)with 633-and 785-nm excitation wavelengths combined with optical clearing(OC)technique was used for ex-vivo study of porcine skin in the Raman fingerprint region.The optical cleari...Confocal Raman microspectroscopy(CRM)with 633-and 785-nm excitation wavelengths combined with optical clearing(OC)technique was used for ex-vivo study of porcine skin in the Raman fingerprint region.The optical clearing has been performed on the skin samples by applying a mixture of glycerol and distilled water and a mixture of glycerol,distilled water and chemical penetration enhancer dimethyl sulfoxide(DMSO)during 30 min and 60 min of treat-ment.It was shown that the combined use of the optical clearing technique and CRM at 633 nm allowed one to preserve the high probing depth,signal-to-noise ratio and spectral resolution simultaneously.Comparing the effect of different optical clearing agents on porcine skin showed that an optical clearing agent containing chemical penetration enhancer provides higher optical clearing efficiency.Also,an increase in treatment time allows to improve the optical clearing efficiency of both optical clearing agents.As a result of optical clearing,the detection of the amide-Ⅲ spectral region indicating well-distinguishable structural differences between the type-Ⅰ and type-Ⅳ collagens has been improved.展开更多
Our previous studies demonstrated the ultrasound-induced skin optical clearing enhancement with topical application of optical clearing agents on in vitro porcine skin and in vivo human skin.The objective of this stud...Our previous studies demonstrated the ultrasound-induced skin optical clearing enhancement with topical application of optical clearing agents on in vitro porcine skin and in vivo human skin.The objective of this study was to investigate the possible mechanisms of the enhanced skin optical clearing by ultrasound medications.Optical clearing effects of ex vivo guinea pig abdomen skin topically applied with 60% glycerol or the combination of 60% glycerol and ultrasound were studied by optical coherence tomography(OCT).Microstructure of skin surface was examined by scanning electron microscopy(SEM).Ultrasound with a frequency of 1MHz and a power of 0.75W over a 3-cm probe was simultaneously applied with glycerol solution for 15min.The combination of 60% glycerol and ultrasound results in a 19% increase in OCT 1/e light penetration depth after 30min,which is much better than 60% glycerol alone.SEM images demonstrated that changes in skin microstructure due to the tight order of the lipid bilayers in the stratum corneum disrupted and the separation of keratinocytes by the application of ultrasound contribute to the ultrasound-enhanced intact skin optical clearing effects.展开更多
The effect of optical cleaning method combined with laser speckle imaging(LSI)was discussed to improve the detection depth of LSI due to high scattering characteristics of skin,which limit its clinical application.A d...The effect of optical cleaning method combined with laser speckle imaging(LSI)was discussed to improve the detection depth of LSI due to high scattering characteristics of skin,which limit its clinical application.A double-layer skin tissue model embedded with a single blood vessel was established,and the Monte Carlo method was used to simulate photon propagation under the action of light-permeating agent.808 nm semiconductor and 632.8 nm He–Ne lasers were selected to study the e®ect of optical clearing agents(OCAs)on photon deposition in tissues.Results show that the photon energy deposition density in the epidermis increases with the amount of tissue°uid replaced by OCA.Compared with glucose solution,polyethylene glycol 400(PEG 400)and glycerol can considerably increase the average penetration depth of photons in the skin tissue,thereby raising the sampling depth of the LSI.After the action of glycerol,PEG 400,and glucose,the average photon penetration depth is increased by 51.78%,51.06%,and 21.51%for 808nm,68.93%,67.94%,and 26.67%for 632.8 nm lasers,respectively.In vivo experiment by dorsal skin chamber proves that glycerol can cause a substantial decrease in blood°ow rate,whereas PEG 400 can signicantly improve the capability of light penetration without a®ecting blood velocity,which exhibits considerable potential in the monitoring of blood°ow in skin tissues.展开更多
In order to enhance the optical clearing effect of topically applied optical clearing agents(OCAs),we evaluated the effect of propylene glycol(PG)as a chemical penetration enhancer(PE)on optical clearing of skin in vi...In order to enhance the optical clearing effect of topically applied optical clearing agents(OCAs),we evaluated the effect of propylene glycol(PG)as a chemical penetration enhancer(PE)on optical clearing of skin in vitro by observation and measurement of optical-transmittance and diffuse-reflectance spectra.Three OCAs,i.e.,glycerol,D-sorbitol and PEG400,and two other penetration enhancers,Azone and Thiazone,were used in this study.The results indicated that the decrease of reduced scattering coefficient caused by OCA/PG was larger than that by pure OCA,and the change by OCA/water was the least after the same treatment time.There were significant differences for the reduced scattering coefficient at 630 nm after 120 min application of agents between OCA and OCA/PG.The efficacy of optical clearing caused by OCA/PG depended on the OCA itself.When PEG400 was mixed with three different PEs,we found the optical clearing were different.The penetration enhancing ability of PG was much better compared to Azone,and suboptimal to Thiazone.Also,this study provides evidence for the use of PG as a PE in order to improve skin optical clearing.展开更多
基金supported by the Research and Development Plan Project of China State Railway Group Co.,Ltd.(Project No.P2023X029).
文摘Purpose–As an important part of the management of railway passenger transport,the rationality and effectiveness of the clearing method of railway passenger transport are directly related to the operating efficiency and service quality of railway passenger transport enterprises.This paper aims to comprehensively and deeply discuss the evolution and development process of China’s railway passenger transport clearing method,analyze its characteristics and influences in each stage,identify the main factors affecting its evolution and development and then put forward thoughts on improving the future development of the clearing method.Design/methodology/approach–Through a detailed review of the railway passenger transport clearing methods from the planned economy period to the reform and opening up period and into the new century,the basis,mode,subject and object of clearing in different development stages are systematically compared.Findings–It comprehensively reveals the evolution of the clearing method,sorted out the characteristics and changes of the clearing method at each stage and the adaptability to the development of railway passenger transport at that time.The characteristics of the development of clearing measures for railway passenger transport in different stages and their far-reaching influence on railway passenger transport business are deeply analyzed.Originality/value–This paper summarized the factors influencing the development of China’s railway passenger transportation clearing approach evolution,including the simplified rules of clearing,enhanced the market adaptability,establishing and perfecting the incentive mechanism,strengthening the construction of informatization,etc.This paper puts forward the ways to improve the railway passenger transportation clearing future development thinking.
基金supported by the National Natural Science Foundation of China(Grant Numbers 62205025 and 61835015)Beijing Natural Science Foundation(7222309)+2 种基金the Open Project Program of Wuhan National Laboratory for Optoelectronics(2020WNLOKF025)CAMS Innovation Fund for Medical Sciences(CIFMS)(2019-I2M-5-061)Beijing Institute of Technology Research Fund Program for Young Scholars(XSQD-202123001).
文摘Vascular-targeted photodynamic therapy(V-PDT)is an effective treatment for port wine stains(PWS).However,repeated treatment is usually needed to achieve optimal treatment outcomes,possibly due to the limited treatment light penetration depth in the PWS lesion.The optical clearing technique can increase light penetration in depth by reducing light scattering.This study aimed to investigate the V-PDT in combination with an optical clearing agent(OCA)for the therapeutic enhancement of V-PDT in the rodent skinfold window chamber model.Vascular responses were closely monitored with laser speckle contrast imaging(LSCI),optical coherence tomography angiography,and stereo microscope before,during,and after the treatment.We further quantitatively demonstrated the effects of V-PDT in combination with OCA on the blood flow and blood vessel size of skin microvasculature.The combination of OCA and V-PDT resulted in significant vascular damage,including vasoconstriction and the reduction of blood flow.Our results indicate the promising potential of OCA for enhancing V-PDT for treating vascular-related diseases,including PWS.
基金supported by the China Agriculture Research System of MOF and MARA(CARS26-05B)the Innovation Research 2035 Pilot Plan of Southwest University,China(SWU-XDPY22002)+1 种基金the Guangxi Science and Technology Planed Project,China(Gui Ke AD23026090)the Guangxi Natural Science Foundation,China(2023GXNSFBA026285).
文摘Citrus yellow vein clearing virus(CYVCV)is a new citrus virus that has become an important factor restricting the development of China’s citrus industry,and the CYVCV coat protein(CP)is associated with viral pathogenicity.In this study,the Eureka lemon zinc finger protein(ZFP)ClDOF3.4 was shown to interact with CYVCV CP in vivo and in vitro.Transient expression of ClDOF3.4 in Eureka lemon induced the expression of salicylic acid(SA)-related and hypersensitive response marker genes,and triggered a reactive oxygen species burst,ion leakage necrosis,and the accumulation of free SA.Furthermore,the CYVCV titer in ClDOF3.4 transgenic Eureka lemon plants was approximately 69.4%that in control plants 6 mon after inoculation,with only mild leaf chlorotic spots observed in those transgenic plants.Taken together,the results indicate that ClDOF3.4 not only interacts with CP but also induces an immune response in Eureka lemon by inducing the SA pathways.This is the first report that ZFP is involved in the immune response of a citrus viral disease,which provides a basis for further study of the molecular mechanism of CYVCV infection.
基金support from the National Key Research and Development Program of China(Grant No.2017YFA0700501),and the Innovation Fund of WNLO.
文摘Three-dimensional(3D)cell cultures have contributed to a variety of biological research fields by filling the gap between monolayers and animal models.The modern optical sectioning microscopic methods make it possible to probe the complexity of 3D cell cultures but are limited by the inherent opaqueness.While tissue optical clearing methods have emerged as powerful tools for investigating whole-mount tissues in 3D,they often have limitations,such as being too harsh for fragile 3D cell cultures,requiring complex handling protocols,or inducing tissue deformation with shrinkage or expansion.To address this issue,we proposed a modified optical clearing method for 3D cell cultures,called MACS-W,which is simple,highly efficient,and morphology-preserving.In our evaluation of MACS-W,we found that it exhibits excellent clearing capability in just 10 min,with minimal deformation,and helps drug evaluation on tumor spheroids.In summary,MACS-W is a fast,minimally-deformative and fluorescence compatible clearing method that has the potential to be widely used in the studies of 3D cell cultures.
基金funded by the National Nature Science Foundation of China(grant nos.62375096,61860206009,32361133552,and 82361138569)the Russian Science Foundation(grant no.24-44-00082)。
文摘Organisms are composed of a vast number of highly specialized cells with characteristic physiological functions determined by their structure.Therefore,understanding physiological and pathological mechanisms requires obtaining 3-dimensional(3D)structural information at the cellular level across tissues,organs,and even the whole body.However,the scattering and absorption of tissue limits the penetration depth of light and thus extremely reduces the imaging performance in deep tissue.It is well known that 3D imaging outperforms 2D imaging in various scenarios for 3D reconstruction of tissue structures.This includes tracing convoluted 3D structures like vasculature,neurons,and prostate glands,as well as elucidating intricate distributions of cells or lesions.
基金Supported by National Natural Science Foundation of China(30770124)~~
文摘[Objective] The research aimed to study a new whole clearing and preparation technology.[Method] After young seeds of Stephania sp.preserved in the alcohol solution,as well as the fresh pistil and nucellus of Sorghum bicolor were separated,they were soaked and cleared in the solution of 84 disinfectant.And the plant materials could be mounted by using three methods.[Result] The globular embryos and heart-shaped embryos at different stages of Stephania sp.as well as the pistil,nucellus,embryo sac,hypostase o...
基金Supported by the Eleventh Five-year Development Planning Project for Instructional Science in Hubei Province (2006B131)~~
文摘MATLAB software and optimal complete subgraph algorithm were used to extract and reveal the microsatellite distribution features in the complete genomes of the tobacco vein clearing virus （NC-003 378.1） from the NCBI database.The results showed that the repetitions number and their location of the N-base group has been extracted and displayed.The largest repetitions of N-base group in the complete genomes of the tobacco vein clearing virus was decreased as the exponential function with the increasing of N.The method used in this study could be applied to the extraction and revealing of the microsatellite distribution features in the complete genomes of other viruses,thereby provided a basis for the research of the structure and the law of function,inheritance and variation by the using of the microsatellite distribution features.
文摘For the first time,the changes in autofluorescence spectra of ex vivo rat skin have been experimentally investigated using the combination of fluorescence spectroscopy and optical immersion clearing.The glucose,glycerol and propylene glycol solutions were used as clearing agents.The optical clearing was performed from the dermal side of skin imitating the in vivo injection of clearing agent under the dermal layers.In this contribution,the common properties of autofluorescence variation during optical immersion clearing were determined.The tendency of autofluorescence signal to decrease with reduction of scattering in tissue was noticed and discussed in detail.However,the differences in the shape of spectral curves under application of different clearing agents showed that optical clearing affects the autofluorescence properties of tissue differently depending on the type of clearing liquid.The results obtained are useful for the understanding of tissue optical clearing mechanisms and for improving techniques such as fluorescence spectroscopy.
基金supported by the National Natural Science Foundation of China(NSFC,Nos.30600126,30700168 and 30900275).
文摘Tissue optical clearing by use of optical clearing agents(OCAs)has been proven to have potential to reduce the highly scattering effect of biological tissues in optical techniques.However,the difference in tissue samples could lead to unreliable results,making it difficult to quantitatively control the dose of OCAs during the course of tissue optical clearing.In this work,in order to study the effects of optical clearing,we customized tissue-like phantoms with optical properties of some biological tissue.Diffuse reflectance and total transmittance of tissue-like phantoms with different OCAs(DMSO or glycerol)and porcine skin tissues were measured.Then optical property parameters were calculated by inverse adding-doubling(IAD)algorithm.Results showed that OCAs could lead to a reduction in scattering of tissue-like phantoms as it did to porcine skin tissue in vitro.Furthermore,a series of relational expressions could be fit to quantitatively describe the relationship between the doses of OCAs and the reduction of scattering effects.Therefore,proper tissue-like phantom could facilitate optical clearing to be used in quantitative control of tissue optical properties,and further promote the application potential of optical clearing to light-based noninvasive diagnostic and therapeutic techniques.
基金supported by the Special Fund for Agro-scientific Research in the Public Interest,China(201203076-05)the National Basic Research Program of China(2014CB138400)
文摘Citrus yellow vein clearing virus （CYVCV） is considered as the causal agent of Citrus yellow vein clearing disease and belongs to the genus Mandarivirus in the family Alphaflexiviridae. Capsid protein （CP） of CYVCV Chongqing isolate （CYVCV- CQ） was produced using a prokaryotic expression system and used as the immunogen for monoclonal antibody （MAb） production. Four highly specific and sensitive murine MAbs and one polyclonal antibody were prepared in this study. Titers of the four MAbs in ascites fluids ranged from 10-6 to 10-7 as determined by indirect enzyme-linked immunosorbent assay （ELISA）. Three serological assays, including dot enzyme-linked immunosorbent assay （dot-ELISA）, tissue blot-ELISA, and double-antibody sandwich （DAS）-ELISA, were developed for quick and reliable detections of CYVCV in citrus samples. The developed dot-ELISA and DAS-ELISA methods could detect CYVCV in the infected citrus leaf crude extracts diluted at 1：2 560 and 1：10 240 （w/v, g mL^-1）, respectively. The detection result of 125 citrus leaf samples collected from citrus groves in Yunnan Province and Chongqing Municipality of China showed that approximately 36% samples were positive for CYVCV. This virus was, however, not＇detected in any sample collected from Zhejiang or Jiangxi Province, China.
基金Supported by National Science and Technology Support Program:Research on unique treatment and principle of inheritance famous old traditional Chinese doctors(No.2013BAI13B021)The Beijing municipal administration of traditional Chinese medicine inheriting program:workstation of famous doctor Zhou Ping-an construction projectThe Innovative Team Project of Beijing University of Chinese Medicine(No.2011-CXTD-08)
文摘OBJECTIVE: To assess the efficacy and safety in 105 patients with seasonal Influenza in Beijing, the mixture prepared with Chinese medicines follows the treatment regimen of releasing exterior cold andclearing interior heat.METHODS: Total 330 patients with seasonal influenza were enrolled and randomly and averagely divided into the Chinese herbal medicine, the western medicine and the Chinese patent medicine group. They were treated with Chinese medicine Oseltamivir Phosphate Capsules and the Scattering Wind and Resolving Toxins Capsules. The main efficacy indicators were the antifebrile onset time and recovery time of body temperature. The efficacy and safety of the mixture was scientifically evaluated. Comparisons of several variables were analyzed.RESULTS: Median antifebrile onset time of the Chinese herbal medicine group was significantly shorter than the western medicine group(P < 0.05) and the Chinese patent medicine group(P < 0.05). The median antifebrile recovery time of the Chinese herbal medicine group was significantly shorter than the Chinese patent medicine group(P <0.05). The groups evaluated by TCM symptom pattern effect, both the Chinese herbal medicine group and Western Medicine group were better than the Chinese patent medicine group(P <0.05). The disappearance rate of main symptoms and some minor symptom patterns of the Chinese herbal medicine group were higher than the other 2 groups.CONCLUSION: The mixture of releasing exterior cold and clearing interior heat could significantly shorten the fever time with safety.
基金the German Research Foundation(DA 2255/1-1,to SCD).
文摘Morphological analyses are key outcome assessments for nerve regeneration studies but are historically limited to tissue sections.Novel optical tissue clearing techniques enabling three-dimensional imaging of entire organs at a subcellular resolution have revolutionized morphological studies of the brain.To extend their applicability to experimental nerve repair studies we adapted these techniques to nerves and their motor and sensory targets in rats.The solvent-based protocols rendered harvested peripheral nerves and their target organs transparent within 24 hours while preserving tissue architecture and fluorescence.The optical clearing was compatible with conventional laboratory techniques,including retrograde labeling studies,and computational image segmentation,providing fast and precise cell quantitation.Further,optically cleared organs enabled three-dimensional morphometry at an unprecedented scale including dermatome-wide innervation studies,tracing of intramuscular nerve branches or mapping of neurovascular networks.Given their wide-ranging applicability,rapid processing times,and low costs,tissue clearing techniques are likely to be a key technology for next-generation nerve repair studies.All procedures were approved by the Hospital for Sick Children’s Laboratory Animal Services Committee(49871/9)on November 9,2019.
文摘Objective： To explore the clinical effect of therapy of clearing hallow viscera in treating critical patients with gastro-enteric function disorder （GEFD）. Methods： Retrospective analysis was carried out on 96 critical patients. They were 48 patients in the treated group treated with Dachengqi Decoction （ 大承气汤） and 48 patients in the control group treated with Western medicine for promoting gastric dynamic force. The recovery rate, recovery time of gastro-enteric function, incidence rate and fatality rate of multiple organ dysfunction syndrome （MODS）, as well as the level of plasma endotoxin （ET） before and after treatment between the two groups were compared. Results： Comparison between the two groups in gastro-enteric function recovery rate （81.3% vs 45.8 %）, functional disorder sustaining time in patients who got recovered （1.2± 0.3 daysvs4.0±1.1 days）, incidence rate （29. 17% vs 52.08%） and fatality rate （28.57% vs 56.00%） of MODS all showed significant difference （ P〈0.05 or P〈0.01 ）. The plasma level of ET after treatment in the treated group was significantly lower than that in the control group （P〈0.05）. Conclusion： Therapy of clearing hallow viscera has a good effect in treating critical patients with gastro-enteric function disorder, and could reduce the incidence and fatality of MODS.
基金supported by the National Natural Science Foundation of China,No.81870655(to MBY)。
文摘Successful establishment of reconnection between retinal ganglion cells and retinorecipient regions in the brain is critical to optic nerve regeneration.However,morphological assessments of retinorecipient regions are limited by the opacity of brain tissue.In this study,we used an innovative tissue cleaning technique combined with retrograde trans-synaptic viral tracing to observe changes in retinorecipient regions connected to retinal ganglion cells in mice after optic nerve injury.Specifically,we performed light-sheet imaging of whole brain tissue after a clearing process.We found that pseudorabies virus 724(PRV724)mostly infected retinal ganglion cells,and that we could use it to retrogradely trace the retinorecipient regions in whole tissue-cleared brains.Unexpectedly,PRV724-traced neurons were more widely distributed compared with data from previous studies.We found that optic nerve injury could selectively modify projections from retinal ganglion cells in the hypothalamic paraventricular nucleus,intergeniculate leaflet,ventral lateral geniculate nucleus,central amygdala,basolateral amygdala,Edinger-Westphal nucleus,and oculomotor nucleus,but not the superior vestibular nucleus,red nucleus,locus coeruleus,gigantocellular reticular nucleus,or facial nerve nucleus.Our findings demonstrate that the tissue clearing technique,combined with retrograde trans-synaptic viral tracing,can be used to objectively and comprehensively evaluate changes in mouse retinorecipient regions that receive projections from retinal ganglion cells after optic nerve injury.Thus,our approach may be useful for future estimations of optic nerve injury and regeneration.
基金This research was supported by Fundamental Research Funds for the CentralUniversities,China(Grant No.XDJK2015D026).Cuihua Liu was the recipient of a scholarship from the China Scholarship Council.Hai Liu,John Hurst,and Michael P.Timko were supported in part by a grant(Award No.GI14732)from 22nd Century Limited,LLC.
文摘Citrus yellow vein clearing virus(CYVCV)is the causal agent of yellow vein clearing disease,a significant and devastating disease of most citrus species including lemon and sour orange.CYVCV,a single-strand positive-sense RNA virus containing six ORFs(Open Reading Frames),represents a new species in the genus Mandarivirus of the Alphaflexiviridae family.The virus can cause particularly serious damage,resulting in reduced tree vigor,lower yields,and decreased marketability of fruit production,and it has been found in India,Turkey,Pakistan,China and Iran.Here we described the geographical distribution of the virus,its transmission mode by grafting,mechanical inoculation,and insects,as well as currently available techniques for detection.In addition,we also discussed practical measures aimed at controlling the disease and provided theoretical guidance to prevent the acquisition and spread of the disease that is a significant step toward ensuring the health of the citrus industry.
基金supported by the National Natural Science Foundation of China(31501611)the Special Fund for Agro-scientific Research in the Public Interest,China (201203076)the Fundamental Research Funds for the Central Universities,China(XDJK2016B021,SWU116012,XDJK2014A001 and XDJK2015A009)
文摘To identity the potential pathogen associated with the yellow vein clearing symptom on lemon trees, the profiles of virus-de- rived small interfering RNAs from citrus samples were obtained and analyzed by deep sequencing method in this study. Twenty-seven contigs almost cover the full length genome of Citrus yellow vein clearing virus （CYVCV） isolate YN were obtained using the small RNA deep sequencing technology. Analysis showed that this isolate CQ shared the highest nucleotide identity with isolate Y1 （JX040635） and YN （KP313242）, both of which belong to the genus Mandarivirus in the family Alphaflexiviridae. Mapping analysis of viral-derived siRNA （vsiRNA） profiles revealed an uneven distribution pattern of their generations along both positive and negative genome strands, and 22- and 21-nt vsiRNAs ranked the majority. BLAST against viroids and other viral databases confirmed that this sample was single-infected only by CYVCV, which indicated that CYVCV was the exact causal agent for the yellow clearing symptom occurring on lemon. This is the first CYVCV isolate detected in Chongqing and the second in China. This result could provide a molecular basis for the investigation of citrus viral diseases to protect citrus health in this region.
基金Ths work was supported by the VEKOP-2.3.2-16-2016-00011 Grantwhich is co-financed by the European Union and European Social FundValery V.Tuchin was supported by RFBR Grant 18-52-16025 and the Grant of the Government of the Russian Federation(Registration No.2020-220-08-2389).
文摘Confocal Raman microspectroscopy(CRM)with 633-and 785-nm excitation wavelengths combined with optical clearing(OC)technique was used for ex-vivo study of porcine skin in the Raman fingerprint region.The optical clearing has been performed on the skin samples by applying a mixture of glycerol and distilled water and a mixture of glycerol,distilled water and chemical penetration enhancer dimethyl sulfoxide(DMSO)during 30 min and 60 min of treat-ment.It was shown that the combined use of the optical clearing technique and CRM at 633 nm allowed one to preserve the high probing depth,signal-to-noise ratio and spectral resolution simultaneously.Comparing the effect of different optical clearing agents on porcine skin showed that an optical clearing agent containing chemical penetration enhancer provides higher optical clearing efficiency.Also,an increase in treatment time allows to improve the optical clearing efficiency of both optical clearing agents.As a result of optical clearing,the detection of the amide-Ⅲ spectral region indicating well-distinguishable structural differences between the type-Ⅰ and type-Ⅳ collagens has been improved.
基金supported by research grants from the National Natural Science Foundation of China(Nos.30470426,30870675).
文摘Our previous studies demonstrated the ultrasound-induced skin optical clearing enhancement with topical application of optical clearing agents on in vitro porcine skin and in vivo human skin.The objective of this study was to investigate the possible mechanisms of the enhanced skin optical clearing by ultrasound medications.Optical clearing effects of ex vivo guinea pig abdomen skin topically applied with 60% glycerol or the combination of 60% glycerol and ultrasound were studied by optical coherence tomography(OCT).Microstructure of skin surface was examined by scanning electron microscopy(SEM).Ultrasound with a frequency of 1MHz and a power of 0.75W over a 3-cm probe was simultaneously applied with glycerol solution for 15min.The combination of 60% glycerol and ultrasound results in a 19% increase in OCT 1/e light penetration depth after 30min,which is much better than 60% glycerol alone.SEM images demonstrated that changes in skin microstructure due to the tight order of the lipid bilayers in the stratum corneum disrupted and the separation of keratinocytes by the application of ultrasound contribute to the ultrasound-enhanced intact skin optical clearing effects.
基金supported by the National Natural Science Foundation of China(Grant No.51727811).
文摘The effect of optical cleaning method combined with laser speckle imaging(LSI)was discussed to improve the detection depth of LSI due to high scattering characteristics of skin,which limit its clinical application.A double-layer skin tissue model embedded with a single blood vessel was established,and the Monte Carlo method was used to simulate photon propagation under the action of light-permeating agent.808 nm semiconductor and 632.8 nm He–Ne lasers were selected to study the e®ect of optical clearing agents(OCAs)on photon deposition in tissues.Results show that the photon energy deposition density in the epidermis increases with the amount of tissue°uid replaced by OCA.Compared with glucose solution,polyethylene glycol 400(PEG 400)and glycerol can considerably increase the average penetration depth of photons in the skin tissue,thereby raising the sampling depth of the LSI.After the action of glycerol,PEG 400,and glucose,the average photon penetration depth is increased by 51.78%,51.06%,and 21.51%for 808nm,68.93%,67.94%,and 26.67%for 632.8 nm lasers,respectively.In vivo experiment by dorsal skin chamber proves that glycerol can cause a substantial decrease in blood°ow rate,whereas PEG 400 can signicantly improve the capability of light penetration without a®ecting blood velocity,which exhibits considerable potential in the monitoring of blood°ow in skin tissues.
基金the National Nature Science Foundation(Grant Nos.30770552 and 30911120074)the National High-Tech Research and Development Program(No.2008AA02Z107)of China.
文摘In order to enhance the optical clearing effect of topically applied optical clearing agents(OCAs),we evaluated the effect of propylene glycol(PG)as a chemical penetration enhancer(PE)on optical clearing of skin in vitro by observation and measurement of optical-transmittance and diffuse-reflectance spectra.Three OCAs,i.e.,glycerol,D-sorbitol and PEG400,and two other penetration enhancers,Azone and Thiazone,were used in this study.The results indicated that the decrease of reduced scattering coefficient caused by OCA/PG was larger than that by pure OCA,and the change by OCA/water was the least after the same treatment time.There were significant differences for the reduced scattering coefficient at 630 nm after 120 min application of agents between OCA and OCA/PG.The efficacy of optical clearing caused by OCA/PG depended on the OCA itself.When PEG400 was mixed with three different PEs,we found the optical clearing were different.The penetration enhancing ability of PG was much better compared to Azone,and suboptimal to Thiazone.Also,this study provides evidence for the use of PG as a PE in order to improve skin optical clearing.
