1.Fluorescent antibody labeling for experimental choroidal neovascularization in mice
Li-ping, GU ; Li, CHEN ; Hui, CHEN ; Jing-sheng, TUO ; Xiao-wei, GAO
Chinese Journal of Experimental Ophthalmology 2011;29(7):619-624
Background Choroidal neovascularization (CNV) is a main cause of visual impairment in many retinal diseases.To create an ideal CNV animal model is very important for the experimental and clinical study of CNV.The assessment method of repeatable and reliable for CNV model is still seldom.Objective This experiment was to explore the label value of fluorescent antibody for visualizing and quantifying the morphologic changes associated with laser-induced CNV.Methods Laser-induced CNV models were created in 30 eyes of 15 male SPF C57BL/6J mice by Krypton red laser irradiating fundus 2 spots around the optical disc with the wavelength 647.1nm,power 260 mW,spot diameter 50μm and exposure time 0.05 seconds.The CNV was evaluated at 5 minutes,4,7,14 and 28 days after laser injury by using fundus photography and fundus fluorescein angiography (FFA),and the successful models were identified as the rupture of Bruch's membrane.The mice were then immediately sacrificed and the eyeballs were enucleated to prepare the choroidal flatmounts.The posterior eye cups were fluorescently labeled with markers of cell nuclei (DAPI,4',6'-diamino-2-phenylindole),endothelial cells (isolectin-B4),and filamentous actin (phalloidin).The CNV areas from specimens were measured by Image pro plus 6.0.Two eyes from one matched mouse without receiving photocoagulation were used as the controlls.This study followed the Standard of Association for Research in Vision and Ophthalmology.Results No any CNV was seen in photocoagulated eyes in 5 minutes and 4 days after laser irradiation.The first sign of CNV appeared at 7 days following photocoagulation.The incidence of fluorescein leakage was 76.47% (26/34),81.81% (18/22),50.00% (5/10) at 7,14 and 28 days,respectively.The fluomicroscope examination showed that in unphotocoagulated areas,retinal pigment epithelial (RPE) cells were visualized with a uniform hexagonal array.Immediately after laser exposure,a circular area devoid of fluorescent labeling was observed,indicating disruption of the choroid-Bruch membrane-RPE complex.On the fourth day,cellular debris and fragmented nuclei were presented and an autofluorescent ring was visible at the site of Bruch's membrane disruption.The number of CNV vessels increased exponentially during the next 3 days.At 7 days,a well-defined isolectin-B4 labeled CNV network was exhibited and lasted for 28 days.The CNV areas were (7.99±0.42)×103μm2,(16.89±3.77)×103μm2,(14.37±4.02)×103μm2 at 7,14 and 48 days after photocoagulation respectively,showing a significant difference among these three groups (F=17.340,P=0.000),and the CNV area was significantly increased in the photocoagulating eyes in 14 days and 28 days compared with 7 days (q=16.46,q=15.54,P<0.01).Conclusion Fluorescent antibody labeling allows the well identification and measurement of laser-induced CNV lesions in mouse choroid/RPE flatmounts.This technique offers excellent morphologic detail and facilitates the study of critical early events in CNV.CNV complexes are labeled at an early stage,providing a more accurate preclinical evaluation of antiangiogenic molecule.
2.Xuebijing injection inhibits the degradation of pulmonary vascular endothelial calyx to improve acute lung injury induced by severe heatstroke
Cheng-Jia DING ; Jing TUO ; Jing-Jing CAO ; Jie-En HUANG ; Chao-Fan LIU ; Hua-Sheng TONG ; Yi CHEN
Medical Journal of Chinese People's Liberation Army 2023;48(12):1387-1394
Objective To investigate the possible mechanism of Xuebijing injection in regulating abnormal degradation of pulmonary vascular endothelial calyx to improve acute lung injury induced by severe heatstroke.Methods Forty-eight Wistar rats were randomly divided into control group,heatstroke group and Xuebijing group.Before heatstroke induction,rats in Xuebijing group were administrated with Xuebijing injection(2 ml/kg,2 times/d)for 3 days.All rats were exposed to an environment with temperature of(40±2)℃and humidity of 65%±5%for 60 minutes to induce heatstroke.Two hours later,the lung wet/dry weight ratio was recorded;the concentration of proteins in BALF was measured;the pulmonary vascular permeability was measured by Evans blue(EB);HE staining was used to observe the pathological changes of lung tissue;the changes of hyaluronic acid(HA)on the surface of pulmonary vessels were observed by immunofluorescence;Western blotting was used to detect the expression of Syndecan-1,Glypican-1,VE-Cadherin,Occludin,VCAM-1 and E-selectin in lung tissues;Enzyme-linked immunosorbent assay(ELISA)was utilized to quantify the concentration of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)in serum and heparanase(HPA)in lung tissue.Results Xuebijing could decrease the lung wet/dry weight ratio,reduce protein exudation and improve pulmonary vascular permeability(P<0.01);reduce the histological injury(P<0.01);reduce the degradation of HA,Syndecan-1 and Glypican-1 on the surface of pulmonary vessels(P<0.01);increase the expression of VE-Cadherin and Occludin(P<0.01);regulate the overexpression of VCAM-1 and E-selectin(P<0.01);down-regulate the expression of TNF-α,IL-6 and HPA(P<0.01).Conclusion Xuebijing injection decrease the expression of HPA,improve the disintegration of pulmonary vascular endothelial calyx,repair the integrity of pulmonary vessels,reduce the damage of cell connections,down-regulate the expression of adhesion molecules,inhibit the inflammatory reaction,relieving acute lung injury caused by severe heat stroke.
3.Distribution characteristics of rs1891385A/C and rs10975519C/T polymorphisms of interleukin-33 gene in Guangxi population.
Jing GUO ; Yang XIANG ; Hua-Tuo HUANG ; You-Fan PENG ; Ye-Sheng WEI ; Yan LAN
Journal of Southern Medical University 2016;36(7):964-968
OBJECTIVETo explore the race- and gender-specific distribution characteristics of rs1891385A/C and rs10975519C/T polymorphism of interleukin-33 (IL-33) gene in Zhuang and Han populations.
METHODSThe polymorphisms of rs1891385A/C and rs10975519C/T of IL-33 gene in 283 subjects from Guangxi Zhuang Autonomous Region were analyzed with single base extension (PCR-SEB) and DNA sequencing to analyze the differences in their distribution frequencies between genders and between Zhuang and Han populations.
RESULTSThree genotypes (AA, AC and CC) were found in rs1891385A/C with frequencies of 64.3%, 32.5% and 3.2%, respectively. The genotype and allele frequencies of rs1891385A/C in this Guangxi population showed no significant difference between Zhuang and Han subpopulations and between genders (P>0.05), but differed significantly from those in European and African black populations (P<0.01). Three genotypes (CC, CT and TT) were identified in rs10975519C/T with frequencies of 34.3%, 53.0%, and 12.7%, respectively, showing no significant ethnic or gender-specific differences in this population (P>0.05). The genotype frequency of rs10975519C/T in this population differed significantly from those in the European and Japanese populations (P<0.01), but the allele frequencies only showed significant differences from those in the European population (P<0.01).
CONCLUSIONrs1891385A/C and rs10975519C/T polymorphisms of IL-33 gene show a race-specific difference.
African Continental Ancestry Group ; genetics ; Asian Continental Ancestry Group ; genetics ; China ; Ethnic Groups ; genetics ; European Continental Ancestry Group ; genetics ; Female ; Gene Frequency ; Genotype ; Humans ; Interleukin-33 ; genetics ; Male ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Polymorphism, Single Nucleotide ; Sequence Analysis, DNA
4.Distribution of single nucleotide polymorphisms of arginine-vasopressin gene in Guangxi healthy population.
Yang XIANG ; Jing GUO ; You-Fan PENG ; Yan LAN ; Hua-Tuo HUANG ; Ye-Sheng WEI
Journal of Southern Medical University 2016;36(7):927-931
OBJECTIVETo study the distribution of single nucleotide polymorphisms (SNP) of arginine-vasopressin (AVP) gene rs66818855 and rs1078152 in Chinese Guangxi healthy population in comparison with that in different ethnic populations.
METHDOSPolymerase chain reaction-single base extension (PCR-SBE) and DNA sequencing were used to detect the allele and genotype frequencies of AVP gene among 303 Chinese healthy individuals in Guangxi, China, and the results were compared with the reported frequencies in 4 other populations (HapMap-CEU, HapMap-YRI, HapMap-JPT, and HapMap-HCB) from Human Genome Project group (HapMap) data.
RESULTSWe found significant AVP gene polymorphisms in this Guangxi healthy population. The frequencies of allele and genotype of AVP gene rs66818855 and rs1078152 polymorphisms in this Guangxi population differed significantly from those in HapMap-CEU population (P<0.01), and allele frequencies of AVP gene rs66818855 polymorphism differed significantly from those in HapMap-YRI populations (P<0.05).
CONCLUSIONThe distribution pattern of AVP gene polymorphisms in this Guangxi population is significantly different from that in other ethnic populations, which might account for the difference in the morbidity of AVP-related disease among different ethnic groups and may have important indications in the study of population genetics and anthropology.
Alleles ; Arginine Vasopressin ; genetics ; Asian Continental Ancestry Group ; China ; Gene Frequency ; Genetics, Population ; Genotype ; Humans ; Polymerase Chain Reaction ; Polymorphism, Single Nucleotide
5.Distribution of polymorphisms at promoter region of miR-17-92 in Guangxi population and association of its polymorphisms and lymphocytes
Rong WANG ; Hai-Mei QIN ; Hua-Tuo HUANG ; Yu-Lan LU ; Jing GUO ; Yan LAN ; Jun-Li WANG ; Ye-Sheng WEI
Chinese Journal of Immunology 2018;34(5):723-726,731
Objective:To investigate the distribution characteristics of polymorphisms of rs9515692C/T and rs1352743A/G in the promoter region of miR-17-92 gene cluster in Guangxi people and compare them with those of other ethnic groups and explore the association of its polymorphisms and lymphocytes.Methods:The rs9515692C/T and rs1352743A/G of miR-17-92 gene cluster were genotyped by using SNaPshot technique and DNA sequencing.Detection of the number of lymphocytes using flow cytometry.The differences of polymorphisms between groups were analyzed statistically.Results:No significant differences of genotype and allele frequency in the two SNPs was observed between different gender in the Guangxi population(P>0.05).However,there were significant differences in the distribution frequencies of genotype and allele of Europeans,Japanese and Africans in rs9515692C/T and rs1352743A/G (P<0.05).Conclusion:Polymorphisms of rs9515692C/T and rs1352743A/G are different in different people.In addition,rs9515692C/T polymorphism may be associated with the number of B cells.
6.Treatment strategies for mass burn casualties.
Jia-ke CHAI ; Zhi-yong SHENG ; Hong-ming YANG ; Dai-feng HAO ; Chuan-an SHEN ; Xiao-ming JIA ; Feng LI ; Sa JING ; Li-gen LI ; Hui-feng SONG ; Chi-yu JIA ; Xiao-ye TUO ; Tian-jun SUN ; Quan HU
Chinese Medical Journal 2009;122(5):525-529
BACKGROUNDMass burn casualties are always a great challenge to a medical team because a large number of seriously injured patients were sent in within a short time. Usually a high mortality is impending. Experiences gained from successful treatment of the victims may be useful in guiding the care of mass casualties in an armed conflict.
METHODSThirty-five burn victims in a single batch, being transferred nonstop by air and highway from a distant province, were admitted 48 hours post-injury. All patients were male with a mean age of (22.4 +/- 8.7) years. The burn extent ranged from 4% to 75% ((13.6 +/- 12.9)%) total body surface area. Among them, thirty-two patients were complicated by moderate and severe inhalation injury, and tracheostomy had been performed in 15 patients. Decompression incisions of burn eschar on extremities were done in 17 cases before transportation. All the thirty-five patients arrived at the destination smoothly via 4-hour airlift and road transportation. Among them, twenty-five patients were in critical condition.
RESULTSThese thirty-five patients were evacuated 6 hours from the scene of the injury, and they were transferred to a local hospital for primary emergency care. The patients were in very poor condition when admitted to our hospital because of the severe injury with delayed and inadequate treatment. Examination of these patients at admission showed that one patient was suffering from sepsis and multiple organ dysfunction syndrome. Dysfunction of the heart, lung, liver, kidney, and coagulation were all found in the patients. Forty-eight operations were performed in the 23 patients during one month together with comprehensive treatment, and the function of various organs was ameliorated after appropriate treatment. All the 35 patients survived.
CONCLUSIONSA well-organized team consisting of several cooperative groups with specified duties is very important. As a whole, the treatment protocol should be individualized, basing on the extent of the injury and the care that the patient had received at the spot. During airlift, the stretchers should be arranged perpendicular to the longitudinal axis of the cabin. The treatment protocol in our hospital consisted mainly of prompt effective relief of all life-threatening complications, followed by early closure of burn wounds, appropriate use of anti-infection therapy, emphasis on nutritional support, correction of metabolic disorders, alleviation of immunosuppression, correction of coagulopathy, and effective support and protection of organ function.
Adolescent ; Adult ; Burns ; drug therapy ; pathology ; surgery ; therapy ; Emergency Medical Services ; Emergency Service, Hospital ; Female ; Hospitals ; Humans ; Male ; Middle Aged ; Time Factors ; Transportation of Patients ; Treatment Outcome ; Young Adult
7.Remote Virtual Companion via Tactile Codes and Voices for The People With Visual Impairment
Song GE ; Xuan-Tuo HUANG ; Yan-Ni LIN ; Yan-Cheng LI ; Wen-Tian DONG ; Wei-Min DANG ; Jing-Jing XU ; Ming YI ; Sheng-Yong XU
Progress in Biochemistry and Biophysics 2024;51(1):158-176
ObjectiveExisting artificial vision devices can be divided into two types: implanted devices and extracorporeal devices, both of which have some disadvantages. The former requires surgical implantation, which may lead to irreversible trauma, while the latter has some defects such as relatively simple instructions, limited application scenarios and relying too much on the judgment of artificial intelligence (AI) to provide enough security. Here we propose a system that has voice interaction and can convert surrounding environment information into tactile commands on head and neck. Compared with existing extracorporeal devices, our device can provide a larger capacity of information and has advantages such as lower cost, lower risk, suitable for a variety of life and work scenarios. MethodsWith the latest remote wireless communication and chip technologies, microelectronic devices, cameras and sensors worn by the user, as well as the huge database and computing power in the cloud, the backend staff can get a full insight into the scenario, environmental parameters and status of the user remotely (for example, across the city) in real time. In the meanwhile, by comparing the cloud database and in-memory database and with the help of AI-assisted recognition and manual analysis, they can quickly develop the most reasonable action plan and send instructions to the user. In addition, the backend staff can provide humanistic care and emotional sustenance through voice dialogs. ResultsThis study originally proposes the concept of “remote virtual companion” and demonstrates the related hardware and software as well as test results. The system can not only achieve basic guide functions, for example, helping a person with visual impairment to shop in supermarkets, find seats at cafes, walk on the streets, construct complex puzzles, and play cards, but also can meet the demand for fast-paced daily tasks such as cycling. ConclusionExperimental results show that this “remote virtual companion” is applicable for various scenarios and demands. It can help blind people with their travels, shopping and entertainment, or accompany the elderlies with their trips, wilderness explorations, and travels.