Establishment and characterization of a canine chondrosarcoma cell line:Mango

In the global progress of bone tumor research,established stable and long-lasting transgenic chondrosarcoma(CSA)cell lines are rare,mainly of murine and human origin,while the establishment of canine CSA cell lines has yet to be reported.This study established a canine CSA cell line to facilitate the basic clinical study of canine CSA.Fifty fve cases of canine osteolytic disease were collected,and more than 10 bone tumor samples from dogs with typical clinical signs were used for primary cell culture.A cell line with stable passaging for more than 100 generations and mouse tumorigenic ability was successfully cultured.According to the clinical characteristics of the dog and the histopathological results of the primary tumor,CSA was diagnosed,and the CSA cell line was designated Mango.Immunohistochemical(IHC)results showed that the immunoreactivity of bone gamma-carboxyglutamate protein(BGLAP),secreted protein acidic and rich in cysteine(SPARC),alkaline phosphatase(ALPL),vimentin(VIM)and S100 were positive.However,the immunoreactivity of pan-cytokeratin(PCK),chromogranin A(CGA),and platelet endothelial cell adhesion molecule-1(CD31)was negative.Immunofuorescence(IF)results showed that the protein expressions in the Mango cell line were consistent with the IHC identifcation of the primary tumor.The Mango cell line’s doubling time was 43.92 h,and the cell formation rate exceeded 20%.There were abnormal chromosome numbers,hetero staining with toluidine blue,and certain calcifcation abilities.It could be passaged stably and continuously without changing the cell morphology and characteristics.In vivo,the cells were successfully injected into the nude mice model with a tumorigenic rate of 100%.The immunophenotype of the xenograft tumor was consistent with that of the primary tumor.Therefore,we efectively established a canine CSA cell line.As a promising cell material,this cell line can be used to construct a tumor-bearing model conducive to the subsequent basic research of canine CSA.Moreover,because of its similarity to human CSA,the animal model of CSA is also indispensable for investigating human CSA.

Molecular characterization of canine parvovirus type 2(CPV2)reveals a high prevalence of the CPV2c genotype among dogs sufering from diarrhea

Canine parvovirus 2(CPV-2)is a highly contagious virus in dogs that typically causes hemorrhagic enteritis and a high mortality rate in unvaccinated puppies.The genetic variability and antigenic diversity of CPV-2 hinder its efective prevention of infection by vaccination.To investigate the epidemiology and genetic characteristics of CPV-2 in China,rectal swabs from afected dogs were collected from diferent animal clinics in Kunshan from 2022 to 2023.Preliminary detection and capsid gene sequencing of CPV-2 were performed using previously described primers and protocols.The overall detection rate for CPV-2 was 16.5%(33/200).A signifcant association was found between the CPV-2-positivity and clinical signs,age,breed and vaccination status.Sequence analysis revealed the presence of CPV-2c genotypes in all positive samples,which were genetically similar to other Asian CPV-2c strains.Notably,four key mutations(A5G,F267Y,Y324I and Q370R)were detected in all isolates,and one novel mutation(I447M)was detected in three CPV-2 isolates.These mutations in the CPV-2 strains could impact vaccine efcacy and the efectiveness of the virus immune evasion.Surprisingly,no recombination events were observed between the identifed CPV-2c strains and reference strains from China.Our data revealed that amino acid residues 324,426 and 440 of VP2 may under strong selection pressure.This pattern of genetic variation in the CPV-2 lineage warrants continuous laboratory-based surveillance programs in other parts of China to better understand the pattern of seasonal distribution and association between emerging genotypes and the intensity of disease severity.

Preparation of anti-canine interleukin-31 receptor alpha polyclonal antibody and evaluation of its therapeutic efect in canine atopic dermatitis

Canine atopic dermatitis(CAD)is a prevalent genetically susceptible infammatory and pruritic allergic skin condition afecting not only the health of dogs but also the quality of life of their owners.Interleukin-31(IL-31)and interleukin-31 receptor alpha(IL-31RA)are essential for the development of pruritus in primates and mice.Hence,it is expected that inhibiting IL-31RA will be an efective approach to alleviate pruritus.The purpose of the study was to produce anti-canine IL-31RA polyclonal antibodies(anti-IL-31RA pAbs)and evaluate their efcacy in inhibiting house dust mite(HDM)-evoked pruritic responses.Dogs were immunized with antigens formed by IL-31RA recombinant short peptides coupled to BSA to produce anti-IL-31RA pAbs.The CAD model was developed by using HDM allergen stimulation,and the efects of IL-31RA pAbs on the reduction of pruritus in CAD model dogs were examined.The Canine Atopic Dermatitis Extent and Severity Index(CADESI)-4 and pruritus Visual Analog Scale(pVAS)were utilized to evaluate pruritic responses,and skin tissue samples were collected from the inguinal area for pathological assessment of skin infammatory cell infltration.The results showed that anti-IL-31RA pAbs with high titers(1:128,000)and specifcity were efectively produced.In the CAD model group,the severity of skin damage,pruritus score,infammatory cell infltration and level of infammatory factors were considerably elevated.Anti-IL-31RA pAbs relieved pruritic behavior and dermatitis in dogs compared to placebo-treated dogs.In conclusion,anti-IL-31RA pAbs efectively suppressed CAD in vivo and are anticipated to be an efective novel treatment for pruritic skin disorders such as CAD.

Update on Canine and Human Rabies in a Rabies Endemic Situation in the Republic of Guinea

Each year, rabies kills nearly 60,000 people worldwide. Asia and Africa are the most affected. It is a public health problem because of the number of stray dogs in these nations. In Guinea, rabies is endemic and cases are often under-reported. The objective of this study was to determine the prevalence of rabies among human and animal exposures in Guinea. This was a retrospective analytical study from 2019 to 2021 conducted in the country’s human and veterinary health divisions and, primarily focused on rabies épidémiologique surveillance. The selection was based on human contact with an animal causing behavioural disorders, as well as animals put under observation following aggression on humans. The data were analysed using Epi Info version 21 software. Qualitative variables were expressed as proportions with 95% CIs, quantitative variables as mean ± standard deviation. The chi2 test and the Fischer exact test were used to compare proportions. A total of 85 veterinary facilities were visited and 100 animals registered, of which 93% were reported cases of exposure to canine rabies. One thousand five hundred and thirty-one (1531) people exposed to rabies were recorded between 2019 and 2021. During the period, the prevalence of human rabies was 0.533% [0.042 - 0.064] at 95% CI and that of canine rabies was 0.55% [0.039 - 0.061] at 95% CI. The biting animal was the dog in 97.2% of cases. Subjects over 45 years of age (57.7%) were the most affected. The prevalence of canine rabies was 0.5% compared with 0.533% for human rabies between 2019 and 2021. This low prevalence does not reflect the true extent of the phenomenon in Guinea, as few people consult health structures in the event of dog bites. Regular surveillance of the human and dog population is necessary for better rabies control in Guinea.

Relationship between Inter Canine Width and Inter Alar Width, Inter Commissural Width, and Inter Canthal Distance: A Pilot Study

The aim of this study was to determine the relationship between inter canine width (ICW) and inter alar width (IAW), inter commissural width (ICoW), and inter canthal distance (ICaD) in Bantu population. Materials and Methods: It was a pilot study included all participants who were aged at least 18 years, without a history of orthodontics treatment from Prosthodontics Department, Faculty of Dental Medicine, University of Kinshasa. The Ethics committee of the National Center of Research for Dental Sciences in DR Congo approved the study protocol (CNRSB 1504.218). The participants medical records were obtained from the interview and dental examination. Nature of the study was explained with participant information sheet, and an informed consent of all participants was obtained. The inclusion criteria were no missing maxillary and mandibular teeth, no diastema, and no anterior restorations, 18 years old before enrollment in the study. The exclusion criteria were inability to meet the above requirements, pregnancy, and refusal to participate in the study. The inter-canine width (ICW), inter alar width (IAW), inter canthal distance (ICaD) and inter-commissural width (ICoW) of each participant were measured with a digital Vernier caliper (Mitutoyo, UK Ltd.,) (0.01 mm) three times and the average value was recorded. The inter canthal distance (ICaD) was measured without applying pressure by bringing the recording parts of the caliper just in contact with the medial angle. The inter alar width (IAW) was marked with a fine marked pencil on the widest outer surface of the alae of the nose on either side or width. Those two points were measured without applying pressure by bringing the recording parts of the caliper just in contact with the outer surface. The participant was told to stop breathing shortly to avoid any change in shape of the nose. The inter-commissural width (ICoW) was determined by measuring the maxillary lip vermilion from commissure to commissure without the application of pressure on the tissue in the relaxed state. The inter-canine width (ICW) was measured indirectly using a dental floss. A dental floss was marked on one end prior to placement in the mouth. Using that point as reference, the dental floss was circumference along the curvature of the anterior dentition such that it passed along the contact point of all the teeth. The distal end of the canine teeth on both sides was then marked on the floss while it was stretched in the patients mouth. Floss was marked on both sides with the marker pencil. The distance between the two proximal contact points was measured and recorded. Data, analysis and validation were performed by the SPSS software (version 22.0, IBM SPSS Statistics, Chicago, IL, USA). Unpaired t-test was used, of Kolmogorov-Smirnov test. Analysis of variance (ANOVA) test was used to test for comparability between socio-demographic characteristics dental measurements. Pearsons correlation coefficients test was calculated to determine the relationship between facial and dental parameters. Significance was set at P 0.01. Results: Of 314 participants enrolled, 202 were included. The age ranged from 18 to 68 years, with a mean age of 40.62 12.99 years. Although the Pearsons correlation coefficients were negative. Ninety-three participants (46%) were men and one hundred and nine (54%) were women. The overall mean age was 40.62 12.99 years. Facial and dental measurements were greater in women than men with significant differences for ICW (p = 0.04). However, no significant difference was seen between men and women for IAW, ICaD and ICoW (p = 0.44, p = 0.23, p = 0.31 respectively). The correlation including Pearsons correlation coefficient and P-values for all participants is not demonstrated. Conclusion: Within the limitations of this study, the results suggest that IAW, ICaD and ICoW cannot be used as a preliminary method for determining the width of the maxillary for anterior teeth for edentulous patients.

Detection of Antinuclear Antibodies in Canine Serum by Immunoperoxidase in MDCK and Indirect Immunofluorescence in HEp2 Cells

Antinuclear antibodies are found in animals suffering from Systemic Lupus Erythematosus (SLE) and some other diseases, their presence in the blood is determined by antinuclear antibody (ANA) test using indirect immunofluorescence (IF) with HEp2 cells as a substrate. In this work, an immunoperoxidase (IP) assay was developed to evaluate the ANAs in canine sera, using canine kidney cell lines (MDCK) and compared with a commercial immunofluorescence test on Hep2 cells for this system, a fluoresceinated anti-canine Ig antibody was standardized. The study was performed on 50 sera from dogs submitted to the laboratory with different clinical diagnoses of autoimmune-associated diseases. The procedures on both cells were unified to perform comparisons of the reactions, direct sera or at different dilutions were added to a monolayer of permeabilized MDCK cells, followed by a peroxidized anti-canine IgG conjugate, and a substrate for the IP reaction. The same sera were tested on the commercial IF assay on Hep2 cell system. In 22/50 cases, the presence of LE cells in peripheral blood was determined. A high correlation was found in the detection of antinuclear antibodies between both cell lines and techniques, however there were differences in the reaction patterns in the nucleus and cytoplasm between cell lines. The diffuse nuclear pattern observed in MDCK cells was more related to the presence of high percentages of LE cells in peripheral blood. The differences found in the results were possibly associated with the presence of homologous antigens between the MDCK cells and the dog. In addition, the methodology and standardization for the use and interpretation of a reference serum was developed to unify the interpretation criteria in the laboratory.

Therapeutic Effects of an Alpha-Casozepine and White Fish Muscle Hydrolyzed Complementary Feed on Canine Separation-Related Disorders and Quality of Life of Dogs and Their Pet Owners

Background: Separation-related disorders are prevalent in dogs and can result in problem behaviours such as inappropriate elimination, destructive behaviour, and distress vocalizations which occur when an owner is absent or perceived as absent. Numerous natural products are marketed for managing anxiety in dogs;however, few, if any, of these products have documented evidence of efficacy in veterinary behaviour. Purpose: This open, multicentric clinical trial aimed to assess the efficacy of a new supplement containing alpha-casozepine and fish muscle hydrolysate on the separation-related problem in dogs and the impact on the quality of life of dogs and their pet owners. Material and Methods: Fifty-one dogs with clinical signs compatible with separation-related problems were included. The dogs were treated with a complementary feed (Zylkene Plus, Vetoquinol, France) for 30 days. Separation-related behaviour assessment (SRP), emotional disorders assessment (EDED) and the quality of life (QoL) of the pets and their pet owner were assessed at baseline and the end of the study by the veterinarians. In addition, owners were asked to rate their dog’s SRP weekly after enrolment using the same questionnaire that the veterinarian used at the beginning of the study. Results: There was a statistically significant improvement in the SRP, EDED, and QoL scores (P Conclusion: The results of this study support the hypothesis that the combination of alpha-casozepine and fish muscle hydrolysate is a promising and effective option to reduce the frequency and severity of separation-related behaviour in dogs and improve the quality of dogs and their owners.

Antiarrhythmic Efficacy of Neferine Assessed by Programmed Electrical Stimulation in a Canine Model of Electropharmacology

An ishemic ventricular tachyarrhythmias canine model was established in open-chest dogs subjected to programmed electrical stimulation (PES)for 5-8 days after acute myocardial infarction. The electrophysiologic effects of neferine (Nef) and procainamide (PA) were observed in this model. With routine methods of PES,ventricular tachycardia (VT)and ventricular fibrillation (VF) could be reproducibly initiated. Both drugs lengthened the QTc interval (P【0.01) and effective refractory period(ERP)of normal and ischemic ventricular myocardia (NERP and IERP) respectively (P【0.01), decreased the dispersion of ERP in ischemic myocardium and the dispersion of ERP in left ventricle (P【0.01), and increased the diastolic excitability threshold of normal and ischemic ventricular myocardia (P【0.01). The two compounds prevented the PES-induced VT or VF (Nef group P【0.01, PA group P【0.05) and ischemia-induced VF (P【0.05). The results indicated that neferine and procainamide may be effective in preventing the onset of reentrant ventricular tachyarrhythmias after myocardial ischemic damage in dogs.

Serological survey on canine coronavirus antibodies in giant pandas by virus neutralization test

In order to survey the infectious situation of canine coronavirus (CCV) in giant panda population, a virus neutralization test detecting specific antibodies against CCV in giant panda抯 sera was established by using two-fold dilutions of serum and 100 TCID50 of the virus. The 62 sera samples of giant pandas, which were gathered from zoos and reserve region of Sichuan Province, China were detected. The neutralization antibody titer of 1:4 was recognized as the positive criterion, 8 sera samples were detected to be positive, and the positive rate was 12.9%. The titers of neutralizing antibody ranged from 1:8 to 1:32. It was the first comprehensive investigation on neutralization antibodies against CCV in giant panda population in China. The results of study showed that the infection of CCV in giant panda population was universal, which has posed a threat to the health of giant panda. Therefore, it is incumbent on us to study safe and effective vaccines to protect giant panda against CCV infection.

Construction of Recombinant Pseudorabies Virus Expressing Canine Distemper Virus H Gene and Analysis on Its Biological Characters

[Objective] The aim was to construct a recombinant pseudorabies virus expressing canine distemper virus H gene and investigate its biological characters.[Method] H gene of canine distemper virus（CDV）strain Onderstepoort was produced by RT-PCR,inserted into pcDNA3.1（＋）vector to construct a expression cassette,which was then subcloned into transfer vector p8AA,prior to the insertion of LacZ expression cassette.The resulting new transfer vector was named as p8AAZH.Subsequently,p8AAZH was co-transfected with the genome of pseudorabies virus（PRV）Bartha-K61 into BHK-21 cells to enable gene recombination and virus package,and the virus solution was collected as cytopathic effect occurring.A series of procedures including blue plaque purification,PCR identification,observation under electron microscope and Western blot were carried out to screen the recombinant pseudorabies virus and identify the protein expression of target gene.Meanwhile,growth curve of the recombinant virus was determined in BHK-21 cells.[Result] The H gene had been inserted into the genome of Bartha-K61 strain,and RPRV-H was the same as Bartha-K61 in the one-step growth curve and cytopathic effect in BHK-21 cells.[Conclusion] The recombinant pseudorabies virus was constructed,and the insertion of H gene did not influence proliferation of recombinant virus,which laid a foundation for development of recombinant canine distemper virus vaccine.

Preparation and Preliminary Identification of Fluorescein Labeled Monoclonal Antibody against Canine Distemper Virus

[Objective] The aim of the present study was to develop a direct immunofluorescence method for the diagnosis of canine distemper （CD） with FITC-conjugated monoclonal antibodies （FITC-McAb）.[ Metbod] The McAb against CDV, designated as CE3, was purified with protein G and labeled with FITC through agitation method. After purification and identification, the optimal working concentration of FITC-labeled CE3 was determined. Then 61 clinical samples of suspected canine distemper were detected by direct immunofluorescence assay. [ Result] The absorption test, blocking test and specificity test showed that the labeled antibody had high specificity and sensitivity, but didn＇t have cross reaction with canine parvovirus （CPV）, canine parainfluenza virus （CPIV）, canine adenovirus （CAV） and rabies virus （RV）. The optimal working concentration was 1：80. The positive rate of clinical suspected samples was 48%. [ Conclusion] The direct immunofluorescence assay developed in this study was rapid, specific and convenient, and had great significance for the early diagnosis of canine distemper.

Three-dimensional Localization of Impacted Canines and Root Resorption Assessment Using Cone Beam Computed Tomography

Summary： The purpose of this study was to develop a new way to localize the impacted canines from three dimensions and to investigate the root resorption of the adjacent teeth by using cone beam computed tomography （CBCT）. Forty-six patients undergoing orthodontic treatments and having impacted canines in Tongii Hospital were examined. The images of CBCT scans were obtained from KaVo 3D exam vision. Angular and linear measurements of the cusp tip and root apex according to the three planes （mid-sagittal, occlusal and frontal） have been taken using the cephalometric tool of the InVivo Dental Anatomage Version 5.1.10. The measurements of the angular and linear coordinates of the maxillary and mandibular canines were obtained. Using this technique the operators could envision the location of the impacted canine according to the three clinical planes. Adjacent teeth root resorption.of 28.26 % was in the upper lateral incisors while 17.39% in upper central incisors, but no lower root resorption was found in our samples. Accurate and reliable localization of the impacted canines could be obtained from the novel analysis system, which offers a better surgical and orthodontic treatment for the patients with impacted canines.

Transcellular transport characteristics of huperzine alone or in combination with ginkgolide B across Caco-2 and Madin-Darby canine kidney cell monolayer

Objective:To study the various processes involved in transcellular transport(TT) of huperzine A alone or in combination with ginkgolide B in Caco-2 and Madin-Darby canine renal(MDCK)cell monolayer.Methods:The transepithelial passage was assayed in the apical-to-basolateral(AP to BL) direction and opposite direction(BL to AP) in both cell lines.The determination of huperzine A and ginkgolide B were performed by high performance liquid chromatography(HPLC).The passage rates of huperzine A and ginkgolide B were calculated.Bi-directional TT(absorption and secretion) were taken in huperzine A and ginkgolide B in Caco-2 and MDCK cell monolayer.Results:TT absorption and secretion kinetics of huperzine A and ginkgolide B across two cells existed at the same time.The passage rates of huperzine A were increased significantly with adding different concentrations of ginkgolide B.Conclusions:The compound preparations of HA in combination with CB for dementia caused by cerebral ischemic have synergistic effects on the pharmacodynamics,and improve the bioavailability through BBB.

Experimental Researches on Carcinogenesis or Tumorigenicity of MDCK Canine Kidney Cell(CKC) Lines and Analysis of Their Chromosome Karyotypes

The chromosomal number variations & structural aberrations of the MDCK cell line, primary feline or canine kidney cell(FKC or CKC) and Hela cell line were investigated and their karyotypes of conventional chromosome bands were analyzed. The carcinogenesis or tumorigenicity testing of these cell lines in about 232 nude mice and for colony formation in soft agarose and for haemagglutination under different concentration of plant lectins of these cells were carried out. Under the prerequisite that the incidence of cancer or tumor in negative-control nude mice inoculated subcutaneously with primary feline or canine kidney cell cultures purified in vitro at passage 3 was 0 (0/22) and 0 (0/10), respectively. The incidence of the progressively-growing malignant tumor(MT) in positive-control nude mice inoculated subcutaneously with Hela cell cultures of KB, X, or NM20/X strain was 10/10, 25/25 and 5/51, respectively. The results showed that the incidence of tumor in nude mice with tetraploid YA strain of MDCK cell during 20 - 45 passages, with hy-podiploid JB strain of MDCK cell on passage 25, with di-and hypoploid JC strain of MDCK cell during 2-15 passages or with hypoploid M strain of MDCK cell during 9 - 27 passages was 28/58, 1/5, 4/18 and 0/31, respectively. The chromosomal analysis results showed that the ratio of difference in the rate of modal chromosome number between high(mcs+ n) and lowest (mcs) passages was not more than 5% - 15% and the structure aberrations was generally 0 - 3% . These results proved that the genetic characteristics of chromosomal number of cell lines determines their tumorigenicity, but it is species-specific. MDCK line has tumorigenicity no matter what its chromosome karyotype is, at least it has very low tumorigenicity even when its modal chromosome number is hypoploid. The repeatedly frozen, thawed and split controls of tumorigenicity-positive cell lines(X strain of Hela, M strain of BHK-21, JA strain of Vero, YA strain of MDCK) have much lower tumorigenicity or are even non-carcinogenesis, and the repeatedly frozen, thawed and split controls of very low tumorigenicity cell lines (M or JC strain of MDCK) are certainly non-carcinogenic and never have increased tumorigenicity. It is thus evident that MDCK cell of M, JB or JC strain can be approved as substrate for the preparation of attenuated viral vaccines, but MDCK cell of YA strain can not be approved as substrate for the preparation of attenuated viral vaccines. In summary, all strains of MDCK cell line have tunorigenicity, at least have low tumor igencity , never have non-cancinogenic MDCK, but very low tumorigenicity MDCK cell strains can certainly be used for the approval production of canine viral vaccines if the DNA content in viral cell cultures was remarkably decreased through conventional means in manufacturing process. Therefore, the master cell stock and working cell bank of MDCK line used for vaccine manufacture were established in China, which are free of infectious agents, and described with respect to cytogenetic characteristics and tumorigenicity.Tests showed that there were correlations among cell line chromosome number variations, anchorage independence in soft a-garose, haemagglutination under plant lectins, and tumor-forming ability in nude mice, thus all the in vitro tests are economic, simple and reliable means for monitoring the tumor-forming ability of MDCK line in nude mice.

Age-related histomorphologic changes in the canine gastrointestinal tract:A histologic and immunohistologic study

AIM： To examine the of the gastric, jejuna physiological aging. changes in the histomorphology and colonic wall of dogs due to METHODS： Full thickness biopsies were taken from the gastrointestinal tracts of 28 dogs of different ages. The thickness of the different layers of the wall was measured and the numbers of proliferating cells as indicated by immunohistochemical detection of Ki67 were counted. RESULTS： In the three excision sites, the thickness of all subepithelial layers increased with rising age. The strongest correlation between age and thickness of the intestinal wall was found in the first 10 years of life and in the jejunum （r =0.6-0.71 for the deep lamina propria mucosa, the muscularis mucosa, and the circular layer of the tunica muscularis）. The number of proliferating cells decreased during aging, with the strongest correlation in the lamina propria mucosa and lamina muscularis mucosa of the jejunum and in the colonic submucosa （r =-0.61 to -0.71）. Epithelial proliferation was only weakly correlated to the age. CONCLUSION： The morphology of the deeper layers and the proliferation of mesenchymal cells of the intestinal wall of healthy dogs are correlated with age. Gastrointestinal epithelial proliferation is only weakly age-correlated.

ELISA Detection of Antibody against Canine Distemper Virus in Fox

Canine distemper is one of the most important common viral epidemics in farmed foxes. In this study, the whole blood samples of foxes that had been immunized with canine distemper vaccine were collected from a farming area in Changli County. The antibody against canine distemper virus in the 85 prepared serum samples were detected with a double-antibody sandwich ELtSA method. The results showed among the 85 samples from the 4 farms, 81 samples were antibody-positive, and only 4 samples had no antibody against canine distemper. The overall antibody level was higher in that farming area. It is indicated canine distemper vaccination and vaccination program have gotten attention from most of the local farmers, and the immune effect of canine distemper vaccine is better.

Pathogenesis of Demyelinating Encephalopathy in Dogs with Spontaneous Acute Canine Distemper

So far, the pathogenesis of demyelination caused by canine distemper virus （CDV） in the central nervous system has remained unclear, although a lot of studies have been done extensively. To further investigate the relation of variety cells in brain to demyelination, this study was performed on 15 dogs with spontaneous acute canine distemper and 2 controls. According to anatomical relation, the brain was divided into cerebrum, cerebral stem and cerebellum. The sections with no, mild, moderate, or severe demyelinating lesions were selected respectively and stained by HE and immunohistochemistry. Immuno-localisation of CDV antigen was used to conftrm CDV infection. The brain was examined for co-localisation of the CDV antigen with either an astrocyte-specific marker, glial fibrillary acidic protein （GFAP）, or an oligodendrocyte-specific marker, galactocerebroside （GalC）. Apoptotic cell was detected by TdT-mediated nick end-labeling assay （TUNEL）. The results demonstrated that the local disturbance of blood circulation mainly included congestion, edema, thrombosis, and disseminated intravascular coagulation （DIC）. The CDV neucleocapsid protein positive reaction, metabolic disorder and apoptosis of oligodendrocytes were observed in demyelinating areas. Lots of astrocytes displayed CDV antigen-positive, especially in their process. Some of them became apoptotic cell confirmed by TUNEL staining. Fibrous astrocytes showed more intense GFAP-positive in mild and moderate demyelinating area. Some of nervous cells located in pyramidal cell layers and nucleus nervi were in degeneration, necrosis. Satellitosis, neuronophagia and apoptotic neurons were examined by hematoxylin and eosin （HE） and TUNEL staining. The results suggested that the demyelinating changes in brain tissues infected with CDV mainly related to the metabolic disorder and apoptosis of ogliodendrocytes and astrocytes; also involved with the local disturbance of blood circulation and some neuron lost.

Pathomorphologic Study of Parasitic Enteritis Caused by Acute Canine Distemper

[Objective]The aim was to survey relationship between acute canine distemper and parasitic enteritis from pathology. [Method]Twelve cases of acute canine distemper with diarrhea were researched as per immunohistochemistry,Haematoxylin Eosin,and PAS staining kit. [Result] Of the twelve diseased dogs （ with diarrhea） ,six were detected caused by coccidium and two were detected by cryptosporidium. Coccidian protozoa is mainly in epithelial cells of jejunum and ileum,and some can be found in cut-off intestinal epithelial cells and in mucus formed by destroyed intesti- nal villus. The most common shapes of coccidian protozoa are trophozoite and schizont. The former is mainly within or among epithelial cells; nucle- us is in center and stained by hematoxylin; protoplasm is in ＂ fined mesh＂ shape. The latter,round or oval,contains much glycogenosome in de- generated intestinal epithelial cells. On the other hand,cryptosporidium is mainly in striated borders of intestinal epithelial cells and intestinal gland cells,leading to destruction of villus and cut-off of cells. Through detection on monoclonal antibody of nucleocapsid proteins of anti-canine distemper virus,it was found that epithelial cells in intestinal mucosa,glandular cells in recesses,lymphocytes and macrophage infittrated in lamina propria and dendritic cells in aggregated nodule were all with positive reactions. [Conclusion]Parasitic diarrhea caused by acute canine distemper occurs when resistance of intestinal mucosa caused by canine distemper virus begins to decline.

Root Resorption of Central Incisor by Impacted Canine in Unusual Transposition: Orthodontics and Implant Rehabilitation: 3-Year Follow-Up

This case presents an unusual transposition of an impacted canine on the central incisor that provides a multidisciplinary approach to its resolution. The patient comes to the consultation for mobility of the central incisor. Clinically, tooth mobility was confirmed with tooth crowding and persistence of the temporary canine. Cone beam tomography (CBCT) observed the atypical position of the permanent canine that caused root resorption of the ipsilateral central incisor and the need to extract both teeth. The conservative treatment of the bone structures during the dental enucleation of the impacted canine carried out by odontosection and the immediate bone grafting of the defect projected an acceptable regeneration of the bone volume for the insertion of the implant. The orthodontic treatment managed to correct the bite, position the upper premolar in the place of the canine tooth and maintain the space to place the implant in the central incisor. The three-year follow-up shows maintenance of the peri-implant crestal bone level, the success of orthodontic treatment with recovery of the patient’s function and aesthetics.

NIH批准对parovirus的专有权

国立心肺及血液研究所(马里兰州)正在物色应用研究B19parovirus《一种潜在的治癌物质和基因治疗的一潜在载体)的搭挡,准备对基础专利“产生B19蛋白的中国仓鼠卵细胞”提供独家专有权.