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Objective To investigate the effect of blue light on the dioptric development of the eyes of lens-induced myopia(LIM)guinea pigs.Methods Three-week-old trichromatic guinea pigs were randomly divided into three groups:control group,white light LIM(WL)group,and blue light LIM(BL)group(420 nm LED light,with an illuminance of 700 lx);guinea pigs in the latter two groups wore-10.00 D lenses in their right eyes to induce myopia.All guinea pigs under-went a 12 h light/12 h dark treatment cycle.Before and 2,4 weeks after the intervention,the diopter,axial length,retinal thickness and choroidal thickness were measured in all groups.After 4 weeks of intervention,the corneal fluorescent stai-ning and retinal Hematoxylin and Eosin(HE)staining were conducted.Results Compared with the control group,from week 0 to week 2 of the intervention(changes in weeks 0-2),the eyes in the WL group drifted(-2.22±1.28)D towards myopia,the axial length lengthened by(0.40±0.05)mm,and the retinal and choroidal thicknesses reduced by(-7.42± 7.04)μm and(-6.29±4.66)μm,respectively;compared with the WL group,in the BL group,the eyes drifted toward hyperopia by(0.48±1.16)D,the axial length increased by(0.20±0.10)mm,and retinal and choroidal thicknesses in-creased by(1.36±7.46)μm and(8.05±8.08)μm,respectively(all P＜0.05).From week 2 to week 4(changes in weeks 2-4),compared with the control group,the diopter in the WL and BL groups progressed towards myopia,with changes of(-4.64±0.50)D and(-2.11±2.02)D,respectively(both P＜0.05);the axial length lengthened,and reti-nal and choroidal thicknesses reduced in the WL group,with changes of(0.44±0.06)mm,(-7.35±5.87)μm and(-4.84±2.61)μm,while the choroidal thickness and the retinal thickness decreased in the BL group,with changes of(-0.33±5.95)μm and(-4.78±4.96)μm,respectively.Observations of corneal fluorescence staining and retinal HE staining indicated that prolonged blue light exposure could lead to damage to corneal and retinal cells.Conclusion Blue light may influence the development of myopia through choroid-related mechanisms,but its inhibitory effect is not positive-ly correlated with time.Prolonged exposure to blue light can damage the cornea and retina,thereby reducing the inhibitory effect.
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Objective:To study the distribution and changes of dopamine transporter (DAT) in guinea pig eyes under different light patterns.Methods:Thirty-six 3-week-old white ordinary-grade guinea pigs were randomly selected and divided into groups of 10 000 lx, 5 000 lx, and 500 lx, with 12 guinea pigs in each group exposed to strong light, medium strong light, and normal light, respectively.Each group was randomly divided into 3 subgroups, with 4 guinea pigs in each subgroup.The 3 subgroups of 500 lx group received light exposure for 5, 20, and 40 minutes, respectively.The 3 subgroups of 5 000 lx group received light exposure for 2, 4, and 40 minutes, respectively.The 3 subgroups of 10 000 lx group received light exposure for 2, 5, and 20 minutes, respectively.After light treatment, each group of guinea pigs was injected with 99mTc-TRODAT-1 for SPECT DAT imaging, and image data were collected by Micro-SPECT.The region of interest (ROI) of guinea pig retinas was analyzed using Micro-CT software.The counts of ROI were expressed as Sum, which reflected the relative distribution or density of DAT.The DAT density between experimental and control eyes of guinea pigs after light exposure, the differences in DAT density between guinea pig eyes under different light intensities, the differences in DAT density between guinea pig eyes after different light durations, and the cumulative and interactive effects of light intensity and light duration on DAT aggregation in guinea pigs were compared.Another 3 guinea pigs were selected, and after light exposure, the 3 guinea pigs' eyes underwent continuous image acquisition for 6 hours at 20-minute intervals, and 18 images per guinea pig were acquired to analyze the trend of DAT density in guinea pig eyes over time.This study was approved by the Ethics Committee of Shanghai General Hospital (No.2020SQ196). Results:The DAT density (Sum value) of experimental eyes at 500, 5 000, and 10 000 lx were 5 598.97±3 159.38, 8 636.78±2 503.16, and 7 407.39±2 053.41, respectively, significantly higher than 4 388.89±2 902.90, 5 981.92±3 057.44, and 5 091.32±2 039.36 of control eyes ( t=5.31, 4.69, 11.80; all at P<0.001). At 500 lx, there was a statistically significant difference in DAT density between the experimental and control eyes of guinea pigs at different light exposure durations ( F=14.01, P<0.01), while no significant difference was found at other light intensities at different light exposure durations (both at P>0.05). When the light exposure time was 5 minutes, the difference in DAT density between the experimental and control eyes of guinea pigs was significantly greater in the 10 000 lx group than in the 500 lx group ( t=-13.22, P<0.001). There was no statistically significant difference between different groups at other light exposure durations (all at P>0.05). No cumulative or interactive effects of light intensity and light duration were found on the differences in DAT density (all at P>0.05). Continuous scanning after illumination showed that DAT density in guinea pig retinas first increased to a peak over time and then gradually returned to normal values. Conclusions:Light, even under moderate or normal light levels, can cause an increase in the secretion of DAT in the retina and stimulate the production of DAT.Light intensity and duration have no cumulative or interactive effects on the distribution and density of retinal DAT.
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Objective:To investigate the alteration of m6A demethylase AlkB homolog 5 (ALKBH5) expression and its impact on form-deprivation myopia (FDM) retina in guinea pigs.Methods:Thirty normal SPF grade 3-week-old tricolor guinea pigs were randomly divided into normal control group and experimental group, with 15 in each group.In the experimental group, the right eyes were covered as FDM group and the left eyes uncovered were set as self-control group.Ocular biometry was performed at one-week intervals from baseline to week 4 of the experiment.Spherical equivalent was detected by streak retinoscopy and axial length was measured by A-scan ultrasonography.Animals were sacrificed after 4 weeks of modeling.The distribution and expression of ALKBH5 protein in the guinea pig retina was detected by immunohistochemical and immunofluorescence staining.Expression of ALKBH5 mRNA and protein in guinea pig retina was detected by real-time fluorescence quantitative PCR and Western blot, respectively.The use of animals in ophthalmic and vision research followed the tenets of Animal Research in Vision and Ophthalmology, and the study was approved by the Ethics Committee of North Sichuan Medical College (No.2023087).Results:At weeks 2, 3, and 4 after myopia induction, diopters and axial lengths were significantly higher in the FDM group than in the normal control group and the self-control group (all at P<0.001). Immunohistochemistry and immunofluorescence assays showed that ALKBH5 protein was expressed in the retinal nerve fiber layer, rod/cone photoreceptor cells, and retinal pigment epithelium (RPE) layer, and was highly expressed in the retinal nerve fiber layer and RPE layer.The relative ALKBH5 immunofluorescence intensity in the normal control group, self-control group and FDM group was 1.000±0.204, 0.874±0.076 and 0.571±0.053, respectively, which was lower in the FDM group than in the normal control and self-control groups, showing statistically significant differences ( t=4.069, P=0.006; t=5.176, P=0.014). After 4 weeks of modeling, ALKBH5 mRNA and protein expressions were significantly lower in FDM group than in normal control and self-control groups (both at P<0.01). Conclusions:The expression of m6A demethylase ALKBH5 is decreased in the retina of FDM guinea pigs, suggesting that ALKBH5 and related m6A methylation modification may be involved in the development and progression of myopia.
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@#Feral pigs are known as triple-threat pests due to their impact on health, agriculture and the ecosystem. Recent emerging infectious disease outbreaks have emphasized their role as reservoirs and amplifiers of disease and highlight the need for increased surveillance in the Western Pacific Region.
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Objective To observe the effects of Jianpi Qutan Huayu Prescription on oxidative stress and inflammatory response in mini-pigs with atherosclerosis(AS);To explore its mechanism based on the NOX5-ERK1/2 signaling pathway.Methods Twelve Bama mini-pigs were randomly divided into control group,model group,and Jianpi Qutan Huayu Prescription low-and high-dosage groups,with 3 pigs in each group.A high-fat diet was used to feed for 24 weeks to construct an AS model,and the treatment group was also supplemented with Jianpi Qutan Huayu Prescription in the feed.The general condition of mini-pigs(body length,abdominal circumference,body mass,food intake,and fecal water content)was measured at week 0,16,and 24 of administration,HE staining was used to observe the morphology of aortic tissue,while oil red O staining was used to observe lipid deposition in aortic and myocardial tissue,transmission electron microscopy was used to observe the ultrastructure of aortic tissue,and a fully automated biochemical analyzer was used to detect serum contents of TC,HDL-C,and LDL-C.ELISA was used to detect the contents of serum reactive oxygen species(ROS),interleukin(IL)-6,IL-10,tumor necrosis factor(TNF)-α,hypersensitivity-C-reactive protein(hs-CRP),vascular cell adhesion molecule-1(VCAM-1),and intercellular adhesion molecule-1(ICAM-1).Western blot was used to detect the expressions of NADPH oxidase 5(NOX5),extracellular signal regulated kinase 1/2(ERK1/2),p-ERK1/2,VCAM-1,and proliferating cell nuclear antigen(PCNA)proteins.Results Compared with the control group,the abdominal circumference,body mass,and food intake of mini-pigs in the model group increased at 16 and 24 weeks(P<0.01),there was significant thickening of the inner membrane of aorta,destruction of endothelial cells,lipid deposition,edema of smooth muscle cells,and significant swelling of mitochondria,serum TC,LDL-C contents and the contents of ROS,IL-6,IL-10,TNF-α,hs-CRP,VCAM-1,and ICAM-1 increased,while the content of HDL-C decreased(P<0.01);the expressions of NOX5,p-ERK1/2,VCAM-1,and PCNA proteins in aortic tissue increased(P<0.01).Compared with the model group,Jianpi Qutan Huayu Prescription low-and high-dosage groups showed a decrease in abdominal circumference,body mass,and food intake at 16 and 24 weeks(P<0.05,P<0.01),the plaque area and lipid deposition were reduced,and the damage to endothelial cells was alleviated,serum TC,LDL-C contents and the contents of ROS,IL-6,IL-10,TNF-α,hs-CRP,ICAM-1,and VCAM-1 decreased,and the content of HDL-C increased(P<0.01,P<0.05);the expressiond of NOX5,p-ERK1/2,VCAM-1,and PCNA proteins in aortic tissue decreased(P<0.01,P<0.05).Conclusion Jianpi Qutan Huayu Prescription can effectively alleviate AS in mini-pigs,and its mechanism may be related to inhibiting the activation of the NOX5-ERK1/2 signaling pathway and alleviating oxidative stress-induced inflammatory response.
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AIM: To avoid the problem of retinal dissection in guinea pig large eyeball tissue sections, different methods were used to optimize the fixation effect of the posterior pole of the eyeball.METHODS: A total of 75 normal guinea pigs(2 weeks old)were randomly divided into 5 large groups. Group A(1-3 small groups), the entire eyeball was fixed with FAS, Davidson fixative 1(D1), and Davidson fixative 2(D2)for 24 h; group B(4-6 small groups), the entire eyeball was fixed with FAS, D1, and D2 for 1 h, then cut the cornea and fix it in their respective fixatives for 2 h; group C(7-9 small groups), the eyeball was fixed in FAS, D1, and D2 for 1 h, divided into left and right halves along the direction of the optic nerve, and then placed them in their respective fixation solutions for 2 h; group D(10-12 small groups), after fixation for 3 h in FAS, D1, and D2, the eyeball was divided into left and right halves along the optic nerve direction; group E(13-15 small groups), the cornea was cut after fixation for 3 h in FAS, D1, and D2. Hematoxylin-eosin(HE)staining was used to compare the fixation effect on posterior eyeball in each group.RESULTS: After fixation, the surface of the eyeballs in groups, 1-6 and 11-15 was smooth and round, with a transparent and bright color. In groups 7-10, the eyeballs were sunken, wrinkled, and deformed. The HE staining showed that the eyeball morphology of groups 1, 5, 6, 14, and 15 was significantly better than the other groups, with a regular internal tissue structure. The eyeballs of the other groups were sunken and wrinkled, and the internal tissue was curled and tangled, with severe retinal detachment. In groups 1, 5, 6, 14, and 15, the retina, choroid, and sclera tissues of group 14 were closely connected, without obvious retinal detachment, rupture, or curling. The tissue structure was clear and visible, and the cells were arranged neatly.CONCLUSION: The fixation effect of cutting the cornea after fixing guinea pig eyeball with D1 fixative for 3 h is the most ideal, and this operation method is simple and suitable for studying the related structures of the posterior pole of the eye.
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Although blood banks based on human blood can provide blood transfusions for the wounded timely and effectively, scientific research has never given up on finding new blood sources due to the restrictions of human blood sources. With the application of transgenic technology and the successful breeding of gene-edited pigs, gene-edited pig blood as a potential source of clinical transfusion has attracted wide attention. Now there are preclinical studies showing the feasibility of transfusing gene-edited pig red blood cells into primates. This paper discusses the related research and future development of xenogeneic transfusion of porcine red blood cells by gene editing.
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Abstract Occurrence of Ureaplasma diversum (U. diversum) has been associated with repro-ductive failures in cattle and detected in pigs with and without pneumonia. However, its rolein the porcine respiratory disease complex (PRDC) is unclear. A cross-sectional study was con-ducted in abattoirs, inspecting 280 pig lungs from eight herds. All the lungs were inspected,processed and classified according to the histopathological analysis. Moreover, bronchoalveolarlavage (BAL) specimens were collected and processed by PCR for detection of U. diversum andMycoplasma hyopneumoniae (M. hyopneumoniae). Ureaplasma sp.---U. diversum and M. hyop-neumoniae were detected in 17.1% and 29.3% of the analyzed BAL specimens, respectively. Theconcomitant presence of both microorganisms was detected in 12.5% of the inspected lungs.Both agents were found in lungs with and without pneumonia. M. hyopneumoniae was detectedin 31.8% of pig lungs with enzootic pneumonia-like lesions, while Ureaplasma sp.---U. diversumwas detected in 27.5% of lungs with these lesions. This descriptive exploratory study providesinformation for future experimental and field-based studies to better define the pathogenicrole of this organism within the PRDC.
Resumen La presencia de Ureaplasma diversum se ha asociado a fallas reproductivas en el ganado bovino y se ha detectado en cerdos con y sin neumonía. Sin embargo, su participación en el complejo de enfermedades respiratorias porcinas (CERP) no es clara. Se llevó a cabo un estudio transversal en matadero, inspeccionando 280 pulmones de cerdo provenientes de ocho piaras. Todos los pulmones fueron inspeccionados, procesados y clasificados según el análisis histopatológico. También se colectaron muestras de lavado broncoalveolar (LBA) y se procesaron mediante PCR para la detección de U. diversum y Mycoplasma hyopneumoniae. Ureaplasma sp.-U. diversum y M. hyopneumoniae se detectaron en el 17,1% y en el 29,3% de los LBA analizados, respectivamente. La presencia concomitante de ambos microorganismos se detectó en el 12,5% de los pulmones inspeccionados. Ambos agentes se encontraron en pulmones con y sin neumonía. M. hyopneumoniae se detectó en el 31,8% de los pulmones con lesiones compatibles con neumonía enzoótica, mientras que Ureaplasma sp.-U. diversum se detectó en el 27,5% de los pulmones con estas lesiones. Este estudio exploratorio descriptivo proporciona información para futuros estudios experimentales y de campo tendentes a definir mejor el papel patógeno de este organismo dentro del CERP.
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Introduction: The available data regarding scalpel and diode laser incisions wound healing is variable, scarce and needs more clarification especially in normal healthy tissues. Hence, this study was aimed to evaluate clinical and histologic parameters of wound healing with scalpel and diode laser on oral mucosa of Guinea pigs. Materials and Methods: This is an experimental study conducted on four male Guinea pigs. Laser and scalpel incisions were made on the right buccal mucosa of each guinea pig approximately 5mm deep and 10 mm in length and both the incisions were placed 10 mm apart using No#11Bard Parker blades and diode laser (640 wavelength) under general anesthesia. Punch biopsies of both the incisions were obtained and compared for clinical changes such as ease of working, intra-operative bleeding, color and approximation of wound edges and histological changes such as epithelial proliferation, inflammation, angiogenesis, and fibrosis on day 1, 3 and 7. Results: Clinically, scalpel wounds showed better healing than laser wounds although laser incisions provided bloodless surgical field. There was no significant difference in inflammation, angiogenesis and fibrosis between scalpel and diode laser incisions on both day 3 and 7 except for the epithelization which was comparatively early and better in scalpel wounds than the diode laser wounds. Conclusion: Diode lasers provide bloodless surgical field with better visibility than scalpel without any difference in oral mucosal wound healing. Considering the clinical benefits, the diode lasers could be a superior alternative to conventional scalpel technique.
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Objective:To investigate an accurate and quantitative method to measure the eyeball morphological parameters of guinea pigs through a method that combines programmed digital techniques and mathematical geometric principles.Methods:Twenty-two three-week-old clean-grade male tricolor guinea pigs were selected and sacrificed by anesthesia overdose.Eyeballs were enucleated.The horizontal and sagittal images of the eyeball were taken with the high-speed photographic model of 13 million pixels macro meter, and the pictures were imported into pycharm programming software.Using the pre-written analysis program of Python 3.9, the conversion coefficient between the photo pixel and the actual length was obtained by a scale, and then the corneal surface was fitted by arc fitting and conic curve fitting.The results of arc fitting were converted to calculate the corneal radius of curvature.The corneal eccentricity was calculated according to the general conic equation (Ax 2+ Bxy+ Cy 2+ Dx+ Ey+ F=0). The corneal asphericity was evaluated by curve fitting between the central 3-mm and the whole cornea.The use and care of the animals complied with Regulations for the Administration of Affairs Concerning Experimental Animals by State Science and Technology Commission.The study protocol was approved by the Ethics Committee of Shanghai Public Health Clinical Center (No.2022-A009-01). Results:The digital method of Python programming can show the corneal contour of guinea pigs completely and clearly.In the transverse plane, there was no significant difference in the corneal curvature measurements among the digital fitting in central 3-mm cornea, digital fitting in whole cornea and curvature meter ( F=1.693, P=0.190). In the sagittal plane, there was a significant difference in the corneal curvature measurements among the three methods ( F=3.500, P=0.030), and the corneal curvature measurements of the whole cornea measured by the curvature meter were significantly greater than those measured by the digital fitting ( P<0.05). There was no statistical significance in the measurements of corneal curvature radius among the three methods in the transverse plane and the sagittal plane ( F=1.817, P=0.170; F=2.050, P=0.133). The horizontal and sagittal corneal eccentricity measured by digital fitting in central 3-mm cornea were 0.55±0.15 and 0.53±0.17, which were lower than 0.66±0.10 and 0.64±0.14 measured by digital fitting in whole cornea, and the differences were statistically significant ( t=-4.860, -5.210; both at P<0.01). Conclusions:It is feasible to use Python programming digital method to measure the corneal curvature and eccentricity of guinea pigs.
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Objective:To observe the effects of blue light intervention on the development of optical defocus-induced myopia in guinea pigs and investigate its underlying mechanisms.Methods:Forty-eight normal-grade two-week-old tricolor guinea pigs were randomly divided into a blue light group and a white light group, with 24 animals in each group.The right eye of guinea pigs was fitted with a -5.00 D lens to establish an optical defocus model as the experimental eye, while the left eye served as the control without any covering.Before the experiment and after 8-week intervention, the refractive power of guinea pigs was measured by streak retinoscopy.The anterior chamber depth, lens thickness, and axial length were measured by A-scan ultrasonography.Corneal curvature radius was determined using a keratometer.After 8-week intervention, the guinea pigs were euthanized through overanesthesia, and the right eyeballs were enucleated and the retinas were isolated.The density of S and M cone cells of the guinea pig retinal sections were observed via immunofluorescence staining.The expression of retinal retinoic acid was assessed by high-performance liquid chromatography.The expressions of retinoic acid receptor (RAR-β) in the retina and matrix metalloproteinase-2 (MMP-2), tissue inhibitor of metalloproteinase-2 (TIMP-2), and type Ⅰ collagen in the sclera were detected by real-time fluorescence quantitative PCR.Changes in scleral thickness were observed through hematoxylin-eosin staining.The use and care of the animals complied with Regulations for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission.The study protocol was approved by the Ethics Committee of Eye and ENT Hospital of Fudan University (No.2022ETKLD10032).Results:After 8 weeks of intervention, guinea pigs in the blue light group showed (0.63±0.12)D of relative hyperopia and a deceleration of axial elongation by (0.08±0.00)mm compared with the white light group in the right eye.In the left eye, guinea pigs in the blue light group showed (0.42±0.09)D of relative hyperopia and a deceleration of axial elongation by (0.08±0.00)mm compared with the white light group.The guinea pigs in blue light group showed (1.52±0.09)D of myopia in the right eye compared with the left eye, with an increase in axial elongation of (0.06±0.00)mm.The guinea pigs in white light group showed (1.66±0.07)D of myopia in the right eye compared with the left eye, with an increase in axial elongation of (0.13±0.00)mm, and the differences were statistically significant (all at P<0.05). The density of M cone cells was lower and density of S cone cells was higher in the blue light group in the dorsal and ventral sides of the retinal sections compared with the white light group, showing statistically significant differences ( t=32.33, 52.23, 42.09, 25.02; all at P<0.05). The deceleration of myopia progression in the blue light group was strongly positively correlated with the increase in S cone cell density on the ventral side ( r=0.95, P<0.01). The expression levels of retinoic acid, RAR-β, and MMP-2 were decreased, and expression levels of TIMP-2 and type Ⅰ collagen were increased in blue light group compared with the white light group, showing statistically significant differences ( t=18.73, 7.45, 3.72, 6.19, 9.03; all at P<0.05). The scleral thickness in the blue light group was (125.0±7.8)μm, which was significantly thicker than (102.0±6.3)μm in the white light group ( t=26.93, P<0.05). Conclusions:Blue light intervention can inhibit the progression of defocus-induced myopia in guinea pigs.Refractive power changes in guinea pigs may be influenced by alterations in retinal cone cell density, retinoic acid expression, and scleral collagen expression.
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Objective To investigate the establishment and optimization of guinea pigs model of cholecystitis,so as to provide the basis for further optimization of experimental design and research on the pathogenesis,treatment and mechanism of cholecystitis.Methods The guinea pigs model of cholecystitis was established by subcutaneous injection of lincomycin hydrochloride,the general condition,B-ultrasound of the gallbladder,blood routine indicators and hepatobiliary histology of the guinea pigs were observed to determine the stabili-ty and application prospect of the cholecystitis model.Results The general state observation showed that guinea pigs in the model group exhibited varying degrees of diet decrease,body weight loss,activity decrease,loose and lusterless hair,listlessness or irritability.B-ul-trasound results showed that,compared with the control group of guinea pigs,with the higher modeling dose,the thickness of the gallblad-der wall were more obvious(P＜0.05).The blood routine index detection found that,with the higher modeling dose,the blood inflamma-tory indicators such as white blood cell count(WBC),lymphocyte(LYM),neutrophil(NEUT),monocyte(MONO)were increased sig-nificantly(P＜0.05).The results of hematoxylin-eosin staining showed that,with the higher modeling dose,pathological changes such as structural changes of gallbladder and liver tissue structure,inflammatory cell infiltration,degeneration and necrosis of liver cells were more obvious.Conclusion In this study,the imaging indicators combined with blood and pathological indicators were used for multi-factor comprehensive evaluation of the modeling effect of cholecystitis and the stability of the inflammatory model,which was more intui-tive,accurate and low-cost than the conventional gallbladder pathological analysis method or combined with enzyme-linked immuno-sorptive assay,and provided a scientific and effective detection index for the evaluation of therapeutic effects of drugs.At the same time,the model with the dose of 120mg/kg lincomycin hydrochloride for guinea pigs cholecystitis was the most stable and difficult to self-heal.
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ObjectiveXenotransplantation is an effective way to address the shortage of human organ donors, but it faces serious immune rejection reactions, including hyperacute rejection caused by blood type differences. Establishing a stable, convenient, and reliable method for pig blood type identification can quickly screen suitable donor pigs for xenograft research.MethodsBanna miniature inbred pigs, Diannan small eared pigs, and Bama Xiang pigs were selected as the research objects. DNA was extracted from the blood, oral buccal mucosa, and fetal fibroblasts of the three strains of pigs using DNA extraction kits. The target fragment of the ABO homologous gene EAA intron 7 in pigs was amplified using PCR method. Blood agglutination reaction was used to detect hemolysis in pig anterior vena cava whole blood after adding anti A and B antibodies. Immunohistochemical method was used to detect the expression level of A antigen in pig heart, liver, spleen, lung, and kidney tissues. Immunofluorescence method was used to detect the expression level of A antigen in pig oral mucosa. By comparing the results of different methods for determining pig blood types, the stability and reliability of the PCR method were verified, and a convenient PCR based pigblood type identification method was established.Results Firstly, the blood PCR results of 69 inbred strains of Banna miniature pigs, 7 Diannan small eared pigs, and 34 Bama Xiang pigs showed 20 AO blood types, 66 AA blood types, and 24 O blood types. The PCR results of fetal fibroblasts from 47 Diannan small eared pigs showed that all 47 fetuses were O blood type. Among them, the oral mucosal PCR results of 8 gene edited cloned pigs were consistent with those of donor fetal fibroblasts, all of which were O blood type. The oral mucosal PCR results of 8 wild-type pigs (2 inbred lines of Banna miniature pigs, 4 Diannan small eared pigs, and 2 Bama Xiang pigs) were consistent with the blood PCR identification results. Then, 11 inbred lines of Banna miniature pigs, 4 Diannan small eared pigs, and 2 Bama Xiang pigs were randomly selected for blood agglutination reaction validation, and the results were consistent with the PCR identification results of both blood samples and oral mucosa samples. Moreover, immuno-histochemical analysis was performed on the heart, liver, lung, kidney, and spleen tissues of one Banna miniature pig inbred line and two Bama Xiang pigs, and the results were consistent with blood PCR identification and blood agglutination reaction results. Finally, oral mucosal samples were collected from 2 inbred strains of Banna miniature pigs and 1 Bama Xiang pig for immunofluorescence detection, and the results were consistent with the blood PCR identification results.ConclusionBy collecting fetal cells and oral mucosal samples from live pigs for PCR detection, the blood type of pigs can be accurately and efficiently identified, providing a convenient method for blood type screening of xenograft donor pigs.
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Aim To discuss the mechanism of Lurong Dabu Decoction on cough variant asthma. Methods Guinea pigs were divided into normal group(CON), model group(OVA), Lurong Dabu Decoction high-dose group(HIGH),low-dose group(LOW), and dexamethasone group(DEX)at random. The CVA model was established by smoking plus injection of OVA, aluminum hydroxide solution and nebulized inhalation to stimulate cough. Gguinea pigs were dissected 24 hours after the last challenge to obtain alveolar lavage fluid(BALF)and lung tissues. Immunoadsorption(ELISA)method was applied to detect the types of inflammatory cells and the content of inflammatory cytokines in BALF; HE and Masson staining of the middle lobe of the left lung were used to observe the pathological changes in lung tissues; immunohistochemical staining was used to observe TLR4 and WNT-5A protein expression and distribution of lung tissues; the protein extracted from the upper lobe of the left lung was used to measure the level of TLR4 and WNT-5A protein in lung tissues by Western blot; immunofluorescence was employed to measure the fluorescence intensity of TLR4 and WNT-5A in lung tissues; flow cytometry was used to detect IL-4 and IFN-γ in guinea pig lung tissues. Results Lurong Dabu Decoction could improve guinea pig airway inflammation, inhibit collagen fiber deposition, reduce the content of IL-4, IL-5, and IL-13 in BALF, and inhibit the protein expression of TLR4 and WNT-5A in lung tissues and increase IFN-γ levels in lung tissues while decreasing IL-4 levels. Conclusion Lurong Dabu Decoction may inhibit the occurrence of CVA through TLR4/WNT-5A signaling pathway.
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Xenotransplantation holds the promise of being used to address the imbalance between organ supply and demand for clinical transplantation. Pigs have natural features that make them more suitable donors for transplant organs than non-human primates. A series of biological barriers that arise after pig organ transplantation have been overcome by genetic engineering and pharmacological suppression. Mean- while, the gradual maturity of the genetic engineering technology has been significantly optimized for suit- able pigs for xenotransplantation, and promoted the development of pig organ transplantation research. Although it will take time for pig organ xenotransplantation to enter the clinical trial stage, recent studies conducted in a few brain-dead or critically ill patients have exhibited the great potential of porcine xeno- transplantation in solving the imbalance between supply and demand of organs for clinical transplantation.
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Objective To investigate the expression levels of scf and c-kit under the regulation of Dahuang Lingxian prescription and the possible mechanism of its effect on gallbladder dynamics, and to provide a theoretical basis for Dahuang Lingxian prescription in preventing the development and recurrence of cholesterol gallstone. Methods A total of 45 specific pathogen-free healthy male guinea pigs were randomly divided into normal group, model group, and traditional Chinese medicine (TCM) group. The guinea pigs in the normal group were fed with normal diet, and those in the model group and the TCM group were fed with high-fat lithogenic diet. After 8 weeks of feeding, 5 guinea pigs were randomly selected from each group, and successful modeling was determined if gallstone was observed with the naked eye in more than 4 guinea pigs. After successful modeling, the guinea pigs in the TCM group were given Dahuang Lingxian prescription by gavage, and those in the model group were given an equal volume of normal saline by gavage. After 8 consecutive weeks of administration by gavage, gallbladder tissue samples were collected, and HE staining was used to observe the pathological changes of gallbladder tissue; Western blot was used to measure the expression level of tumor necrosis factor-α (TNF-α) in gallbladder tissue; immunohistochemistry was used to measure the protein expression levels of scf and c-kit in gallbladder smooth muscle tissue. A one-way analysis of variance was used for comparison of normally distributed continuous data between multiple groups, and the least significant difference multiple comparison method was used for further comparison between two groups. Results HE staining showed marked inflammation of gallbladder tissue in the model group, and compared with the model group, the TCM group had a significantly lower degree of inflammation. Western blot showed that the model group had the highest expression level of TNF-α in gallbladder tissue, followed by the TCM group and the normal group ( P < 0.05); immunohistochemistry showed that compared with the model group, the normal group and the TCM group had significantly higher protein expression levels of scf and c-kit in gallbladder smooth muscle tissue ( P < 0.05). Conclusion Dahuang Lingxian prescription can enhance the dynamic function of the gallbladder, possibly by upregulating the scf/c-kit signaling pathway in interstitial cells of Cajal in gallbladder.
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Background & objectives: Japanese encephalitis (JE) is a major public health problem in India. The first outbreaks of JE have been reported from the North-eastern regions of Assam, particularly from the Lakhimpur district of Assam between July–August 1989. In Assam every year many people died due to JE. This study was performed to investigate the molecular epidemiology of JE in pigs in Lakhimpur district of Assam and the risk factors associated with causing Japanese encephalitis in pigs. This study will help to map out the endemic regions and to know where and when to apply the most control strategies towards the prevention and control of the disease. Methods: A total of 342 serum samples from pigs were collected from 10 organized and 20 unorganized farms from 9 blocks and recorded to age, sex and breed and tested by RT-PCR. Pig farms and the surrounding environment were studied for assessment of farm-level risk factors responsible for JEV infection in pigs. Results: Out of 342 samples tested for detection of the E gene of JEV, 14 samples were found to be positive with a prevalence rate of 4.09%. Age, sex and breed-wise higher cases were found in at the age group above 12 months, sex wise female and breed-wise local pigs. Pig farms less than 500 meters from risk factors like rice field, stagnant water source, wild bird exposure to farm and mosquito exposure at farm/ bite to pigs, found to be more numbers of JE cases. Interpretation & conclusion: Molecular epidemiology of JE in pigs, and humans; positive at Lakhimpur recommend the need for uninterrupted surveillance of this virus in pigs specially those areas where pig population is more and all risk factors are present.
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Resumen Trichophyton benhamiae es un dermatofito zoofílico. Puede causar tinea corporis, tinea faciei y tinea capitis. Se caracteriza por producir lesiones inflamatorias, sobre todo en niños. El objetivo de esta publicación es describir 7 casos clínicos de pacientes pediátricos atendidos entre julio del 2019 y enero del 2020 en nuestra institución. A los pacientes se les solicitó estudio micológico convencional, con posterior confirmación con MALDI-TOF MS y secuencia-ción del ADN ribosomal. Se aisló e identificó T. benhamiae como agente etiológico; el nexo epidemiológico fue el contacto con cobayos. Estas son las primeras descripciones de infecciones causadas por T. benhamiae en Argentina. Al realizar estudios micológicos convencionales, este agente puede confundirse con otros dermatofitos, por lo tanto, se requieren herramientas como MALDI-TOF MS o la secuenciación para llegar a un diagnóstico definitivo. Es importante contar con datos epidemiológicos, como el contacto con mascotas no tradicionales, para una presunción diagnóstica adecuada.
Trichophyton benhamiae is a zoonotic dermatophyte that can cause tinea corporis, tinea faciei and tinea capitis, producing inflammatory lesions, especially in children. In this publication, we describe 7clinical cases of pediatric patients that occurred in our institution between July 2019 and January 2020. All patients underwent a conventional mycological study. The identification of fungi isolates was confirmed by MALDI-TOF MS and sequencing of the ribosomal DNA. T. benhamiae was identified as the etiological agent, whose epidemiological link in all cases was the contact with Guinea pigs. This is the first description of infections caused by T. benhamiae in Argentina. This dermatophyte can be misidentified as other more frequent dermatophytes when performing conventional studies. Molecular technology should be used to reach a definitive diagnosis. It is important to have epidemiological data from patients such as contact with non-traditional pets, especially Guinea pigs, for an adequate presumptive diagnosis of this dermatophytosis.
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Abstract Introduction: Electrocochleography has recently emerged as a diagnostic tool in cochlear implant surgery, purposing hearing preservation and optimal electrode positioning. Objective: In this experimental study, extra-cochlear potentials were obtained during cochlear implant surgery in guinea pigs. The aim was to determine electrophysiological changes indicating cochlear trauma after cochleostomy and after electrode implantation in different insertion depths. Methods: Normal-hearing guinea pigs (n = 14) were implanted uni- or bilaterally with a multichannel electrode. The extra-cochlear cochlear nerve action potentials were obtained in response to acoustic stimuli at specific frequencies before and after cochleostomy, and after introduction of the electrode bundle. After the electrophysiological experiments, the guinea pigs were euthanized and microtomography was performed, in order to determine the position of the electrode and to calculate of the depth of insertion. Based on the changes of amplitude and thresholds in relation to the stimulus frequency, the electrophysiological data and the position obtained by the microtomography reconstruction were compared. Results: Cochleostomy promoted a small electrophysiological impact, while electrode insertion caused changes in the amplitude of extra-cochlear electrophysiological potentials over a wide range of frequencies, especially in the deepest insertions. There was, however, preservation of the electrical response to low frequency stimuli in most cases, indicating a limited auditory impact in the intraoperative evaluation. The mean insertion depth of the apical electrodes was 5339.56 μm (±306.45 - 6 inserted contacts) and 4447.75 μm (±290.23 - 5 inserted contacts). Conclusions: The main electrophysiological changes observed during surgical procedures occurred during implantation of the electrode, especially the deepest insertions, whereas the cochleostomy disturbed the potentials to a lesser extent. While hearing loss was often observed apical to the cochlear implant, it was possible to preserve low frequencies after insertion. © 2020 Associação Brasileira de Otorrinolaringologia e Cirurgia Cérvico-Facial. Published by Elsevier Editora Ltda. This is an open access article under the CC BY license (http:// creativecommons.org/licenses/by/4.0/).
Resumo Introdução: A eletrococleografia surgiu recentemente como uma ferramenta diagnóstica na cirurgia de implante coclear, objetiva a preservação da audição e o posicionamento ideal dos eletrodos. Objetivo: Determinar as alterações eletrofisiológicas indicativas de trauma coclear após a cocleostomia e após o implante do eletrodo em diferentes profundidades de inserção. Método: Neste estudo experimental, potenciais extracocleares foram obtidos durante a cirurgia de implante coclear em cobaias. Cobaias com audição normal (n = 14) foram implantadas uni- ou bilateralmente com eletrodo multicanal. Os potenciais de ação do nervo coclear extracoclear foram obtidos em resposta a estímulos acústicos em frequências específicas antes e após a cocleostomia e após a introdução do feixe de eletrodos. Após os experimentos eletrofisiológicos, as cobaias foram submetidas à eutanásia e a microtomografia foi feita para determinar a posição do eletrodo e calcular a profundidade de inserção. Com base nas mudanças de amplitude e limiares em relação à frequência do estímulo, os dados eletrofisiológicos e a posição obtida na reconstrução microtomográfica foram comparados. Resultados: A cocleostomia promoveu um pequeno impacto eletrofisiológico, enquanto a inserção do eletrodo causou alterações na amplitude dos potenciais eletrofisiológicos extra-cocleares em uma ampla faixa de frequências, especialmente nas inserções mais profundas. Houve, entretanto, preservação da resposta elétrica aos estímulos de baixa frequência na maioria dos casos, indicou um impacto auditivo limitado na avaliação intraoperatória. A profundidade média de inserção dos eletrodos apicais foi 5339,56 μm (± 306,45 - 6 contatos inseridos) e 4447,75 μm (± 290,23 - 5 contatos inseridos). Conclusão: As principais alterações eletrofisiológicas observadas durante os procedimentos cirúrgicos ocorreram durante o implante do eletrodo, especialmente nas inserções mais profundas, enquanto a cocleostomia alterou os potenciais em menor grau. Embora a perda auditiva seja frequentemente observada em posição apical ao implante coclear, foi possível preservar as baixas frequências após a inserção.
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Background: Commensal microflora such as Escherichia coli and Enterococcus spp. are representative indicators of antimicrobial resistance (AMR) as they are part of the normal intestinal microflora and can acquire and disseminate AMR to pathogenic or zoonotic bacteria like Salmonella spp. Objective: To investigate the state of AMR among E. coli and Salmonella spp., potential pathogens in humans, isolated from cecal contents of pigs submitted to a veterinary diagnostic laboratory in Colombia from 2016 to 2019. Methods: Susceptibility testing was conducted using the Kirby-Bauer disk diffusion method according to the Clinical and Laboratory Standards Institute guidelines for antimicrobial zone diameter breakpoints. An E. coli strain (ATCC 25922) was used as the quality control organism. Isolates showing resistance to three or more antimicrobial classes were classified as multidrug-resistant (MDR) as defined by a joint group of the European Centre for Disease prevention and Control and the Center for Disease Control and Prevention of the USA. Results: A total of 112 E. coli and 192 Salmonella spp. colonies were isolated from 557 samples received between 2016 and 2019. In order of decreasing frequency, E. coli was resistant to tetracycline (100%), sulfamethoxazol-trimethoprim (97.5%), amoxicillin (86.4%), enrofloxacin (82.6%), tylosin (82.1%), doxycycline (59%), neomycin (50%), ciprofloxacin (45.5%), ceftiofur (35%), gentamicin (30%), tilmicosin (29%), and fosfomycin (12.5%). When compared with E. coli, Salmonella spp. was generally resistant to the same agents with slightly less resistance (between 10-30%) to eight of the antimicrobials tested. Salmonella spp. showed <20% resistance to three antimicrobials, as follows: neomycin (17%), gentamicin (16%), and fosfomycin (14%). Multi-resistance occurred in 68.7% (77/112) of E. coli and 70.3% (135/192) of Salmonella spp. isolates. Resistance of Salmonella spp. was alarming to all the critically important antimicrobials tested: fluoroquinolones (enrofloxacin, ciprofloxacin), ceftiofur (third- generation cephalosporin), and macrolides (tylosin). Conclusions: According to our results, there is a high level of multi- drug resistance (MDR) in E. coli and Salmonella spp. It is necessary to implement a nationwide antimicrobial resistance monitoring program in Colombia, together with proper antimicrobial prescribing guidelines for pigs. The indiscriminate use of antimicrobial growth promoters by the swine industry is generating widespread bacterial resistance and should be discontinued.
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Antecedentes: Flora comensal como espécies de Escherichia coli e Enterococcus são tipicamente escolhidas como indicadores representativos de la resistência antimicrobiana (AMR), pois fazem parte da flora intestinal normal e podem adquirir e disseminar AMR a bactérias patogênicas ou zoonóticas como Salmonella spp. Objetivo: Investigar o estado da AMR entre E. coli e Salmonella spp. isolados do conteúdo cecal de porcos colombianos submetidos ao Laboratório de Diagnóstico Veterinário de 2016 a 2019, ambos sendo patógenos potenciais em humanos. Métodos: O teste de suscetibilidade foi conduzido usando o método de difusão em disco Kirby-Bauer de acordo com as diretrizes do Instituto de Padrões Clínicos e Laboratoriais para pontos de quebra de diâmetro da zona antimicrobiana. A cepa de E. coli (ATCC 25922) foi usada como organismo de controle de qualidade. Os isolados que apresentam resistência a três ou mais classes de antimicrobianos foram classificados como multirresistentes (MDR), conforme definido por um grupo conjunto do Centro Europeu para Prevenção e Controle de Doenças e Centro para Controle e Prevenção de Doenças dos EUA. Resultados: Um total de 112 E. coli e 192 Salmonella spp. foram isolados de 557 amostras submetidas entre 2016 e 2019. Em ordem decrescente de frequência, a resistência a E. coli foi: tetraciclina (100%), sulfametoxazol-trimetoprim (97,5%), amoxicilina (86,4%), enrofloxacina (82,6%), tilosina (82,1%), doxiciclina (59%), neomicina (50%), ciprofloxacina (45,5%), ceftiofur (35%), gentamicina (30%), tilmicosina (29%) e fosfomicina (12,5%). Quando comparada com E. coli, Salmonella spp. foi geralmente resistente aos mesmos agentes com resistência ligeiramente menor (entre 10-30%) a oito dos antimicrobianos. Apenas três antimicrobianos apresentaram resistência a Salmonella spp. abaixo de 20% da seguinte forma: neomicina (17%), gentamicina (16%) e fosfomicina (14%). Multi-resistência ocorreu em 68,7% (77/112) de E. coli e 70,3% (135/192) de Salmonella spp. isolados. Resistência de Salmonella spp. foi alarmante para todos os antimicrobianos criticamente importantes testados: fluoroquinolonas (enrofloxacina, ciprofloxacina), ceftiofur (cefalosporina de terceira geração) e macrolídeos (tilosina). Conclusões: Esses resultados indicam um alto nível de resistência a múltiplos medicamentos (MDR) e que um Programa Nacional de Monitoramento da Resistência Antimicrobiana é necessário para a Colômbia, juntamente com a implementação de diretrizes de prescrição de antimicrobianos para suínos. O uso indiscriminado de antimicrobianos para promoção de crescimento na indústria suína está claramente promovendo resistência generalizada e deve ser interrompido.