Mastitis is one of the most widespread infectious diseases that adversely affects the profitability of the dairy industry worldwide. Accurate diagnosis and identification of pathogens early to cull infected animals and minimize the spread of infection in herds is critical for improving treatment effects and dairy farm welfare. The major pathogens causing mastitis and pathogenesis are assessed first. The most recent and advanced strategies for detecting mastitis, including genomics and proteomics approaches, are then evaluated .Finally, the advantages and disadvantages of each technique, potential research directions, and future perspectives are reported. This review provides a theoretical basis to help veterinarians select the most sensitive, specific, and cost-effective approach for detecting bovine mastitis early.

Objective:To detect the characteristics of vascular remodeling after carotid balloon injury model in rats using ultrasound biomicroscopy(UBM), and to discuss the application value of UBM technique by comparing ultrasonic characteristics with histopathological results.Methods:Carotid balloon injury was performed in 10-week-old SD rats(11 female and 11 male) by 2F Fogarty balloon catheter. The left common carotid artery(CCA) was injured and the right side in the same animal was used as an uninjured control. Arterial structures and hemodynamics were evaluated pre-procedure and post-procedure at 7, 14 days.The intima-media thickness(IMT) inner diameter, outer diameter, lumen area, vessel area, peak systolic velocity, end diastolic velocity of CCA were measured by UBM, and the vascular resistance index, shear stress and blood flow were calculated to evaluate the vascular hemodynamics. The histological data were obtained by H&E staining in cross-sections at 14 days after balloon injury. The characteristics of arterial structure and hemodynamic changes at various time points were compared, the structural changes of CCA between injured and control side after injury were compared. The Spearman correlation and linear regression were used to test the correlation between ultrasonic and histological measurements 14 days after balloon injury.Results:①Compared with pre-procedure, the IMT at 14 days after balloon injury was increased, the inner diameter was decreased, the shear stress in ultrasound was increased(all P<0.05). H&E staining histological test showed that IMT and neointima area in male rats were larger than those of female rats (all P<0.001). ②After carotid balloon injury, the lumen area decreased, but the CCA underwent compensatory positive remodeling and the vessel area increased. ③Significant correlations were demonstrated between UBM and histology in IMT, inner diameter, outer diameter and vessel area of CCA( rs=0.819, 0.965, 0.896, 0.955; all P<0.001). The vessel area value measured by UBM was larger than that of histology( P=0.006). Conclusions:The CCA of rats can be showed clearly by UBM in males and females. The arterial structure cab be measured by UBM accurately with good correlation with histology, as did arterial hemodynamic parameters, which may be benefit for the study in carotid balloon injury model of rats.

Objective:To explore the clinical and CT imaging features of immune checkpoint inhibitor-associated pneumonia (CIP) and to improve the early diagnostic ability of CIP.Methods:From June 1, 2020 to October 31, 2021, the clinical data and chest CT images of 2 067 patients with advanced malignant tumor treated with immune checkpoint inhibitor (ICI) in the First Medical Center, Chinese PLA General Hospital were retrospectively analyzed. Patients with CIP were enrolled according to the guidelines for CIP diagnosis, and the incidence, time from the start of medication to the onset of CIP, medication cycle, imaging features, imaging patterns, CT grade and outcomes were analyzed. χ 2 test was used to compare the incidence of CIP in patients with or without basic lung disease. Results:Among 2 067 patients with malignant tumors treated with ICI, 67 patients developed CIP, the incidence of CIP was 3.2%. The incidence of CIP was significantly different between 386 patients with basic lung disease (7.00%, 27/386) and 1 681 patients without basic lung disease (2.4%, 40/1 681) (χ 2=21.32, P<0.001). The time from the start of medication to the onset of CIP was 7-367 d (median 52 days), and the duration of medication was 1-12 cycles (median 2 cycles). The imaging features of CIP presented as ground glass opacities in 54 cases (80.6%), solid nodules in 26 cases (38.8%), consolidations in 25 cases (37.3%) and irregular reticular opacities in 24 cases (35.8%). The main radiologic pattern was organizing pneumonia (OP, 34 cases, 50.7%), and followed by diffuse alveolar damage (DAD) pattern (14 cases, 20.9%). According to CT grading, there were 26 cases in low risk grade, 17 cases in moderate risk grade and 24 cases in high risk grade. Of 43 low-and medium-risk grade cases, 25 were OP pattern, accounting for 58.1%, and among 24 high-risk grade patients, 13 were DAD pattern, accounting for 54.2%. Forty-three of the 52 patients were initially untreated, of which 23 patients progressed, 17 had lesion shrinkage, and 3 had resolution, and relapsed in 8 cases after resolution or drug withdrawal. Conclusions:The imaging manifestations of CIP are mainly ground glass opacities, nodules, consolidations, and irregular reticular opacities. The radiologic patterns are mainly OP and DAD. OP is the most common pattern in low-moderate risk grade CIP and DAD is the most common pattern in high risk grade CIP. Patients with basic lung disease are more likely to get CIP.

As a dioxygenase, Ten-Eleven Translocation 2 (TET2) catalyzes subsequent steps of 5-methylcytosine (5mC) oxidation. TET2 plays a critical role in the self-renewal, proliferation, and differentiation of hematopoietic stem cells, but its impact on mature hematopoietic cells is not well-characterized. Here we show that Tet2 plays an essential role in osteoclastogenesis. Deletion of Tet2 impairs the differentiation of osteoclast precursor cells (macrophages) and their maturation into bone-resorbing osteoclasts in vitro. Furthermore, Tet2 mice exhibit mild osteopetrosis, accompanied by decreased number of osteoclasts in vivo. Tet2 loss in macrophages results in the altered expression of a set of genes implicated in osteoclast differentiation, such as Cebpa, Mafb, and Nfkbiz. Tet2 deletion also leads to a genome-wide alteration in the level of 5-hydroxymethylcytosine (5hmC) and altered expression of a specific subset of macrophage genes associated with osteoclast differentiation. Furthermore, Tet2 interacts with Runx1 and negatively modulates its transcriptional activity. Our studies demonstrate a novel molecular mechanism controlling osteoclast differentiation and function by Tet2, that is, through interactions with Runx1 and the maintenance of genomic 5hmC. Targeting Tet2 and its pathway could be a potential therapeutic strategy for the prevention and treatment of abnormal bone mass caused by the deregulation of osteoclast activities.
5-Methylcytosine ; analogs & derivatives ; chemistry ; metabolism ; Animals ; Cell Differentiation ; Cells, Cultured ; Core Binding Factor Alpha 2 Subunit ; genetics ; metabolism ; DNA-Binding Proteins ; physiology ; Genome ; Genomics ; Mice ; Mice, Knockout ; Osteoclasts ; cytology ; metabolism ; Proto-Oncogene Proteins ; physiology

BACKGROUND:Previous studies have confirmed that ethanol can promote adipogenic differentiation of bone marrow mesenchymal stem cells (BMSCs), and up-regulate the expression of PPARγ and aP2 in the tumor necrosis factor α (TNF-α) signaling pathway. As a member of the ZIP protein family, Zrt/Irt-like protein 1 (ZIP1) is closely related to bone metabolism and osteogenic differentiation.OBJECTIVE:To study the effect of BMSCs transfected by ZIP1 on TNF-α signaling pathway in the process of adipogenic differentiation.METHODS:The BMSCs from rabbits were isolated and cultured under different concentrations of alcohol (0.03, 0.09,0.15, 0.21 mol/L), followed by transfection by ZIP1 siRNA and ZIP1 expression vector.RESULTS AND CONCLUSION:After culture in alcohol, the expression levels of aP2 and PPARγ proteins were both significantly increased (P < 0.05), and the level of triglyceride was increased in all alcohol groups except for 0.03 mol/L alcohol group (P < 0.05). After siRNA transfection, the expression levels of aP2 and PPARγ as well as the level of triglyceride were increased significantly in all the alcohol groups (P < 0.05); however, ZIP1 transfection decreased the expression levels of aP2 and PPARγ proteins (P < 0.05). To conclude, ZIP1 siRNA could promote the adipogenic differentiation of BMSCs through the activation of TNF-α signaling pathway.

BACKGROUND: With the increasing number of patients undergoing knee arthroplasty in China, the complications caused by the mismatch of prosthesis with anatomical parameters arouse extensive attentions.OBJECTIVE: To measure the anatomical parameters of the knee joints in healthy Zhuang ethnic group from Guangxi Zhuang Autonomous Region, China, using three-dimensional (3D) reconstruction technology, thus providing reference for the design and placement of the hip prosthesis.METHODS: Sixty-eight individuals, including 30 females and 38 males, were selected from 217 healthy people undergoing CT examination. All data were imported into Mimics 15.0 software used for 3D reconstruction, and then the knee parameters were then measured and analyzed.RESULTS AND CONCLUSION: There was no significant difference in the parameters of the bilateral femur and tibia (P > 0.05). The femoral parameters had significant differences between genders (P < 0.05). The width of tibial platform, sagittal length of lateral tibial platform, distance between medial tibial plateau and fibular head, and posterior slope of tibial plateau showed no significant differences between genders (P > 0.05), while there were significant differences in the width of medial/lateral tibial platform, and sagittal length of medial tibial platform between genders (P < 0.05). The sagittal length of medial femoral condyle was negatively correlated with age, and all femoral parameters were positively correlated with height; while only the width of medial and lateral tibial platform were positively correlated with height. To conclude, 3D reconstruction technology is available for research on the morphology of the knee joint in the Zhuang ethnic group from Guangxi Zhuang Autonomous Region, which provides references for prosthesis design and research appropriate for the Zhuang ethnic group.

Objective To explore the severe fever with thrombocytopenia syndrome bunyavirus (SFTSV) existence time in the body,and the correlation between viral load and the severity and prognosis of disease.Methods The clinical data of 125 SFTS patients from May 2015 to October 2016 in Weihai Central Hospital in Shandong province were analyzed retrospectively.Patients were divided into low viral load group and high viral load group according to the SFTSV RNA levels.Neurological symptoms,bleeding tendency,the incidence of myocardial damage and severe pneumonia,laboratory biochemical index and prognosis of two groups were compared.SFTSV RNA of 46 cases were detected dynamically.Data with homogeneity of variance were tested by t test,and data with heterogeneity of variance was tested by rank sum test.Results Among the 125 cases,64 were male and 61 were female.The mean age was (59.0±3.6) years old.One hundred and one cases were cured,and 24 died.SFTSV RNA loads in low viral load group(81 cases) were (3.08± 1.01) copies/mL,and those in high viral load group (44 cases) were (5.69 ± 0.99) copies/mL,with statically significant difference (t =11.78,P＜0.05).By the dynamic detection of SFTSV RNA load in 46 patients,viral loads in most patients were gradually declined after 1 week of onset,and cleared after 23 days.The incidence of neurological symptoms,bleeding tendency,severe myocardial damage and pneumonia of two groups showed significant difference (x2 =92.987,38.711,75.889 and 54.680,respectively,all P＜0.05).The viral loads of patients who died varied from 1.06× 104 copies/mL to 5.78 × 107 copies/mL.White blood counts of two groups showed no significant difference (t =0.181,P＞ 0.05).The platelet counts of two groups had significant difference (t =2.869,P＜0.05).AST and γ-GT of two groups also had significant difference (P＜0.01 and 0.05,respectively).creatine kinase,creatine kinase isoenzyme,lactic dehydrogenase and hydroxybutyrate dehydrogenase of two groups all had significant difference (P＜0.01 or 0.05).Serum sodium,blood calcium and glucose of the two groups had significant difference (P＜0.01 or 0.05).activated partial thromboplastin time of the two groups showed significant difference as well (t=5.623,P＜0.01).Conclusions After the onset of SFTSV infection,the virus existence in the body may less than 4 weeks.Viral loads are closely associated with disease severity and prognosis.The higher the viral loads are,the heavier organ dysfunction could be and the higher mortality is.

BACKGROUND:Endothelial progenitor cells have good prospects for clinical application;especial y as seed cells, they are involved in construction of tissue engineered blood vessels and disease treatment. OBJECTIVE:To investigate the biological characteristics of endothelial progenitor cells derived from granulocyte colony-stimulating factor (G-CSF) mobilized peripheral blood. METHODS:Mononuclear cells from G-CSF mobilized peripheral blood (n=9) and normal adult peripheral blood (n=8) were obtained and cultured in expanded medium. The immunophenotype of endothelial progenitor cells was investigated by FACS and immunohistochemistry. Real-time PCR was used to detect the expression of different cytokines. Proliferation and adhesion ability of endothelial progenitor cells were detected by MTT assay and adhesion assay. Moreover, the abilities of vasculogenesis in vitro and Dil-labeled acetylated low density lipoprotein uptake were detected. The acquired cells were seeded onto the basilar membrane gel containing vascular endothelial growth factor and basic fibroblast growth factor to induce angiogenesis. RESULTS AND CONCLUSION:Endothelial progenitor cells derived from G-CSF mobilized peripheral blood were similar to those from normal adult peripheral blood in phenotype and morphology. FACSs and immunohistochemistry showed that endothelial progenitor cells were positive for endothelial cellmarkers, such as CD31, vWF, CD34, FLK-1, VE-Cadherin and CD133. In addition, the expression of vascular endothelial growth factor and stromal cel-derived factor 1 were significantly increased in G-CSF mobilized endothelial progenitor cells. Moreover, endothelial progenitor cells derived from two sources possessed the abilities of angiogenesis in vitro and Dil-labeled acetylated low density lipoprotein uptake. But, the number and the proliferation ability of endothelial progenitor cells derived from G-CSF mobilized peripheral blood were better than that of endothelial progenitor cells derived from normal adult peripheral blood. These findings indicate that there is a population of cells with characteristics of endothelial progenitor cells in G-CSF mobilized peripheral blood. They possess the abilities of vasculogenesis in vitro. Moreover, compared to endothelial progenitor cells derived from normal adult peripheral blood, we could obtain more endothelial progenitor cells in G-CSF mobilized peripheral blood and those cells show better proliferation ability.

BACKGROUND:Bone marrow mesenchymal stem cells derived from multiple myeloma patients are characterized by pluripotential differentiation, immunoregulation and supporting hematopoiesis, but whether these features are affected after cryopreservation is unclear.
OBJECTIVE:To study the biological characteristics of cryopreserved bone marrow mesenthymal stem cellderived from multiple myeloma patients.
METHODS:Bone marrow mesenchymal stem cells derived from multiple myeloma patients were cryopreserved in-196 ℃ liquid nitrogen for 1 month (short term) and 12 months (long term) with Iscove’s modified Dulbecco’s medium containing 10%dimethyl sulfoxide and 40%fetal bovine serum as cryoprotectant. The viability and proliferation ability of thawed bone marrow mesenchymal stem cells were investigated. Hematopoiesis support of thawed bone marrow mesenchymal stem cells in vitro was detected by long-term bone marrow culture and the methylcellulose progenitor assay. The immunoregulatory ability of thawed mesenchymal stem cells was detected by mixed lymphocyte culture assay.
RESULTS AND CONCLUSION:The cellviability was (92.9±7.5)%and (86.7±9.2)%for mesenchymal stem cells cryopreserved as long as 1 month or 12 months, respectively. Furthermore, thawed bone marrow mesenchymal stem cells possessed the ability of supporting colony forming and could significantly suppress proliferation of T lymphocytes. At last, there were no changes detected as compared with pre-cryopreserved mesenchymal stem cells in the abilities of proliferation, hematopoiesis support and immunoregulation. Cryopreservation can decrease the cellviability of bone marrow mesenchymal stem cells derived from the patients with multiple myeloma, but cannot affect the abilities of proliferation, hematopoiesis support and immunoregulation.

Objective To study the immunogenicity,immune modulatory effects and mechanisms of mesenthymal stem ceils (MSCs) derived from the bone marrow of patients with multiple myeloma (MM).Methods The mononuclear cells from the bone marrow of MM patients were obtained and cultured.Immunophenotypes were investigated by using FACS.The levels of cytokines were determined by using enzyme linked immunosorbant assay (ELISA).The endocytosis of monocyte-derived dendritic cells (DCs) was investigated by using FACS.Moreover,the immunoregulatory ability of DCs on proliferation of T lymphocytes was detected by using mixed lymphocyte culture assay.Results MM-derived MSCs had no expression of HLA-DR and costirnulatory molecules (CD40,CD80,CD83 and CD86).MM-derived MSCs could significantly suppress proliferation of T lymphocytes in a dose-dependent manner,which could be reversed by antiTGF-β1 and anti-HGF antibodies.MM-derived MSCs inhibit the endocytosis of DCs.MM-derived MSCs could inhibit the secretion of IL-12 and significantly inhibit the function of DCs on proliferation of T lymphocytes.Conclusion MM-derived MSCs harbored low immunogenicity and immunoregulatory effect in vitro,and this effect was achieved through cytokines.