The Comparison of PCR Kits for the Detection of Erythrocytic Parasites on Filter Paper.

Dried blood spot (DBS) based PCR was considered an inexpensive and feasible method for detecting pathogens in the blood. The DBS carrier filter paper and PCR kits are crucial for accurate diagnosis. We evaluated 4 types of filter papers and 20 PCR kits for DBS samples. The PCR detecting Plasmodium results showed that the minimum detection limit of the 4 filter papers was 1 × 102 parasites/µL, and the positive rates of 20 PCR kits ranged from 0% to 100%. PCR results were satisfactory for detecting Plasmodium falciparum (P. falciparum) and Plasmodium. vivax (P. vivax) in archived DBS samples and Babesia gibsoni (B. gibsoni) in fresh pet DBS samples. Our results provided a useful reference for the detection of blood pathogens with DBS samples and direct PCR, especially for screening the cost-efficacy combination of filter paper and PCR kit in resource-limited areas.

Radiography, CT, and MRI Diagnosis of Enzootic Nasal Tumor in Goats Infected With Enzootic Nasal Tumor Virus.

The aim of this study was to describe radiography, computed tomography (CT), and magnetic resonance imaging (MRI) findings of enzootic nasal tumors in goats infected with enzootic nasal tumor viruses. Five of six goats with a mean age of 2 years, showed clinical signs of respiratory disease. Head radiographs showed increased density of the unilateral or bilateral nasal cavity in four goats, and a CT scan showed that the space-occupying lesion of the nasal cavity originated from the ethmoid bone and was enhanced homogeneously postcontrast in all goats. The nasal concha was destroyed and the paranasal sinus mucosa was thickened and filled with fluid in some goats. On MRI, the mass exhibited equal or slightly higher signal intensity on T2 weighted images, equal signal intensity on T1 weighted images, a high signal on fluid-attenuated inversion recovery images and heterogeneous enhancement postcontrast. After dissection, histopathological examination of the mass and virus genome detection of the nasal secretions confirmed that the intranasal mass was a low-grade adenocarcinoma and that the goats were infected with enzootic nasal tumor virus type 2. In conclusion, CT and MRI have high diagnostic values for enzootic nasal tumors because they match the postmortem findings and are more accurate than radiography.

ABCC9 Is Downregulated and Prone to Microsatellite Instability on ABCC9tetra in Canine Breast Cancer.

Tumorigenesis is associated with metabolic abnormalities and genomic instability. Microsatellite mutations, including microsatellite instability (MSI) and loss of heterozygosity (LOH), are associated with the functional impairment of some tumor-related genes. To investigate the role of MSI and LOH in sporadic breast tumors in canines, 22 tumors DNA samples and their adjacent normal tissues were evaluated using polyacrylamide gel electrophoresis and silver staining for 58 microsatellites. Quantitative real-time polymerase chain reaction, promoter methylation analysis and immunohistochemical staining were used to quantify gene expression. The results revealed that a total of 14 tumors (6 benign tumors and 8 breast cancers) exhibited instability as MSI-Low tumors. Most of the microsatellite loci possessed a single occurrence of mutations. The maximum number of MSI mutations on loci was observed in tumors with a lower degree of differentiation. Among the unstable markers, FH2060 (4/22), ABCC9tetra (4/22) and SCN11A (6/22) were high-frequency mutation sites, whereas FH2060 was a high-frequency LOH site (4/22). The ABCC9tetra locus was mutated only in cancerous tissue, although it was excluded by transcription. The corresponding genes and proteins were significantly downregulated in malignant tissues, particularly in tumors with MSI. Furthermore, the promoter methylation results of the adenosine triphosphate binding cassette subfamily C member 9 (ABCC9) showed that there was a high level of methylation in breast tissues, but only one case showed a significant elevation compared with the control. In conclusion, MSI-Low or MSI-Stable is characteristic of most sporadic mammary tumors. Genes associated with tumorigenesis are more likely to develop MSI. ABCC9 protein and transcription abnormalities may be associated with ABCC9tetra instability.

Why the anti-nodal quasiparticle dispersion is so flat in the superconducting cuprates?

The emergence of the coherent quasiparticle peak and the development of the peak-dip-hump structure in the anti-nodal region below T c is the most prominent non-BCS signature of the under-doped high-T c cuprates, in which no coherent quasiparticle can be defined in the anti-nodal region above T c. The peak-dip-hump structure has been commonly interpreted as the result of the coupling of the electron to some Bosonic mode. However, such an electron-Boson coupling picture does not answer the question of why the quasiparticle dispersion is so flat in the anti-nodal region, a behavior totally unexpected for Bogoliubov quasiparticle in a d-wave BCS superconductor. Here we show that the sharp quasiparticle peak in the anti-nodal region should be understood as a new pole in the electron Green's function generated by the strong coupling of the electron to diffusive spin fluctuation around the antiferromagnetic wave vector Q = (π, π), rather than a nearly free Bogoliubov quasiparticle in a d-wave BCS superconductor. More specifically, we find that the normal self-energy of the electron from the scattering with the diffusive spin fluctuation manifests itself mainly as a level repulsion effect and is responsible for the reduction of both the quasiparticle dispersion and the quasiparticle dissipation rate in the anti-nodal region. We argue that the peak-dip separation in the anti-nodal spectrum should not be interpreted as the energy of the pairing glue.

Microsatellite Instability and MMR Genes Abnormalities in Canine Mammary Gland Tumors.

Early diagnosis of mammary gland tumors is a challenging task in animals, especially in unspayed dogs. Hence, this study investigated the role of microsatellite instability (MSI), MMR gene mRNA transcript levels and SNPs of MMR genes in canine mammary gland tumors (CMT). A total of 77 microsatellite (MS) markers in 23 primary CMT were selected from four breeds of dogs. The results revealed that 11 out of 77 MS markers were unstable and showed MSI in all the tumors (at least at one locus), while the other markers were stable. Compared to the other markers, the ABC9TETRA, MEPIA, 9A5, SCNA11 and FJL25 markers showed higher frequencies of instability. All CMT demonstrated MSI, with eight tumors presenting MSI-H. The RT-qPCR results revealed significant upregulation of the mRNA levels of cMSH3, cMLH1, and cPMSI, but downregulation of cMSH2 compared to the levels in the control group. Moreover, single nucleotide polymorphisms (SNPs) were observed in the cMSH2 gene in four exons, i.e., 2, 6, 15, and 16. In conclusion, MSI, overexpression of MMR genes and SNPs in the MMR gene are associated with CMT and could be served as diagnostic biomarkers for CMT in the future.

The driving mechanism of the d-wave orbital order in the iron-based superconductors.

We study the driving mechanism and the form of the orbital order in the electronic nematic phase of the iron-based superconductors (IBSs) within the random phase approximation of a 5-band model. We find the magnetic correlation energy of the system can be significantly improved when an orbital order of the d-wave form is spontaneously generated. On the other hand, the magnetic correlation energy increases as one introduce either an on-site or an extended s-wave orbital order. More specifically, we find that the on-site orbital order is disfavored by the Hund's rule coupling and the extended s-wave orbital order is disfavored by the stripy magnetic correlation pattern in the IBSs. Our work indicates that the orbital order and the spin nematic order in the IBSs develop in a cooperative fashion in the electronic nematic phase of the IBSs and should be both understood as a component of a composite order.

Sequence analysis of the thrombospondin-related adhesive protein gene and heat shock protein 70 gene of Babesia gibsoni isolated from dogs in Nanjing, China.

In this study, the thrombospondin-related adhesive protein (TRAP) gene and the heat shock protein 70 (Hsp70) gene of Babesia gibsoni isolated from the naturally infected dog in the Nanjing area were cloned and sequenced. Twenty blood samples were collected from the suspected cases of babesiosis at the animal hospital of Nanjing Agriculture University. Genomic DNA was extracted from the blood samples, and the 18S rDNAs were amplified by PCR to confirm Babesia infection. As a result, 10 cases of Babesia 18S rDNA gene amplification were positive in the 20 blood samples, confirming that the 10 cases were infected with canine Babesia. The TRAP and the Hsp70 gene fragments were amplified from all 10 positive cases. The four isolates, named NJN1, NJN2, NJN3 and NJN4, were sequenced and compared with other isolates in Asian. The similarity of TRAP and Hsp70 gene sequences among four isolates in Nanjing were above 99%. The Nanjing isolates were closely related to isolates from Taiwan and Japan. Indian isolates were different form Chinese and Japanese isolates, despite the very high similarity of the 18s rRNA genes sequence. These results suggest that the TRAP and Hsp70 genes have a reference value for the genetic diversity analysis of Babesia gibsoni.

Copper-modified palygorskite is effective in preventing and treating diarrhea caused by Salmonella typhimurium.

The aim of this research was to develop effective alternative therapies to reduce antibiotic use in animal agriculture. In this study, the efficacy of copper-modified palygorskite (CM-Pal) in preventing diarrhea caused by Salmonella was specifically examined both in vitro and in vivo. The CM-Pal was prepared with palygorskite (Pal) and copper nitrate. The antibacterial activity of the CM-Pal was detected by comparing the differences in cell numbers on plate count agar before and after adding the CM-Pal to Salmonella typhimurium cultures. Seventy ICR mice were then allocated into seven groups. Five groups (the treatment groups) were infected with S. typhimurium by intraperitoneal (i.p.) injection and were given Pal, CM-Pal, montmorillonite powder, gentamicin, and physiological saline, respectively. One group (the prevention group) was given CM-Pal before infection with S. typhimurium. Another group (the uninfected group) was not infected with S. typhimurium. The effects of Pal, CM-Pal, montmorillonite powder, and gentamicin on the treatment or prevention of diarrhea in the mice were examined by stool studies, fecal scoring, and assessment of growth performance and villus height. The CM-Pal had satisfactory anti-bacterial properties in vitro: the antibacterial rate was 100% after 2 h incubation with S. typhimurium NJS1 cultures (1×106 colony-forming units (CFU)/ml). In the in vivo experiment, the CM-Pal exerted superior effects in the treatment and prevention of diarrhea in mice compared with Pal, montmorillonite powder, and gentamicin. In the CM-Pal group, no mice showed signs of diarrhea at 24 h post infection (p.i.), and all mice fully recovered from infection. However, the Pal group, montmorillonite group, and gentamicin group only recovered after 48, 48, and 96 h, respectively. The villus height level in the CM-Pal treatment group recovered at 3 d p.i. However, the recovery time of the other groups was longer (at least 5 d). The CM-Pal prevention group had a better effect on weight gain than the other groups. This study suggested that CM-Pal may be an effective alternative to conventional antibiotics for the treatment and prevention of animal diarrhea caused by Salmonella.

Altered expression of the mismatch repair genes in DF-1 cells infected with the avian leukosis virus subgroup A.

The absence or deficiency of DNA mismatch repair (MMR) activity results in microsatellite instability (MSI) in cancer. The avian leukosis virus (ALV) causes neoplastic disease in chickens. In this study, the status of MMR, MSI, the cell cycle and apoptosis were detected in DF-1 cells after avian leukosis virus subgroup A infection. Flow cytometry analysis results indicated that there was no significant difference in cell apoptosis between the control and infected groups. The percentage of cells in S and G2 phases were increased in the infected group. MSI and mutation of MSH2 and MLH1 gene exons were absent in DF-1 cells after infection. Levels of MSH2 and MLH1 mRNA were dramatically increased in DF-1 cells after infection. These results demonstrated that ALV RAV-1 infection may promote the expression of MSH2 and MLH1 genes rather than resulting in gene mutations. Mismatch repair functions were normal and may be have relationships with the arrest of S phase and G2 phase.

Canine Babesiosis in China Caused by Babesia gibsoni: A Molecular Approach.

BACKGROUND: To provide a point of reference to study the epidemiology and clinical expression of canine babesiosis in China. METHODS: A total of 30 dogs infected with canine babesiosis were evaluated by mean of clinical history, physical examination, hematological, restriction fragment length polymorphism of PCR products (PCR-RFLP) and sequencing analysis. RESULT: The most prevalent clinical abnormalities were lethargy (100%), anorexia (100%), pale or icteric mucous membranes (80%), fever (70%) and dark urine (70%). Hematology parameters revealed that anemia and thrombocytopenia were the major abnormalities in blood of dogs infected with canine babesia. The results of PCR-RFLP and sequencing analysis indicated that B. gibsoni was the main species responsible for canine babesiosis cases at the time of the study in Nanjing, China. CONCLUSIONS: The results provide valuable information for better understanding of the epidemiology of canine babesiosis in China.

Microsatellite instability in chicken lymphoma induced by gallid herpesvirus 2.

Microsatellite instability (MSI) has been found in a range of human tumors, and little is known of the links between MSI and herpesvirus. In order to investigate the relationship between MSI and Gallid herpesvirus 2 (GaHV-2)-induced lymphoma, fifteen Marek's disease (MD) lymphomas were analyzed through using 46 microsatellite markers, which were amplified by PCR from DNA specimens of lymphoma and normal muscular tissues from the same chicken. PCR products were evaluated by denaturing polyacrylamide gel electrophoresis for MSI analysis. MSI was proved in all lymphomas, at least in one locus. Thirty of the 46 microsatellite markers had microsatellite alterations. These results suggested that GaHV-2-induced lymphoma in chickens is related to MSI, and this is the first report to demonstrate that MSI is associated with the GaHV-2 induced lymphoma in chicken.

[From meridians and acupoints to self-supervision and control system: a hypothesis of the 10th functional system based on anatomical studies of digitized virtual human].

Chinese acupuncture and moxibustion has been widely accepted as a useful therapeutics all over the world, but its mechanism has not been fully defined. For this purpose, a reticular framework of whole-body fascia and connective tissues has been established by means of digitized virtual human technique. The virtual acupoints represented three-dimensionally were compared with the sites for stimulation in practice of traditional Chinese medicine (TCM) acupuncture therapy. The results showed that the fascial network constituted by the connective tissues may be the anatomical basis for acupuncture therapy. We found that the acupoints were mainly located where thick connective tissues were present. In this fascial network, sensitive nerve endings, active cells and lymphatic vessels abounded in the sites with thick connective tissue, and needling at these sites induced definite biological effects. In light of biological phylogeny and embryo development, we believe that the connective tissue network may constitute a new functional system in the human body, the Self-supervision and control system (Fasciology), which provides a theoretical base for acupuncture therapy.