Objective To evaluate the feasibility of imaging the rat cardiac conduction system (CCS) using transaortic antegrade perfusion of Alexa Fluor 633-labeled antibodies targeting hyperpolarization-activated cyclic nucleotide-gated cation channel 4 (HCN4) and connexin (Cx). The study also sought to optimize antibody dosage, perfusion duration, and assess the photostability of the dye. Methods Ex vivo rat heart model with transaortic antegrade perfusion was established using 33 male SPF-grade Sprague-Dawley (SD) rats. Primary and secondary antibody solutions were sequentially perfused in an antegrade manner. After perfusion for predetermined durations, the atrioventricular junction was observed, and the fluorescence intensity of the corresponding area was recorded. Five dose-gradient groups (n=3 rats/group), five perfusion time-gradient groups (n=3 rats/group), and ten continuous LED light exposure time-gradient groups (using 3 rats prepared with a fixed dose and time) were established to observe and record regional fluorescence intensity. Standard immunofluorescence staining was performed on both paraffin and frozen sections for comparative histological analysis. Results A region of aggregated red fluorescent signal was observed in the atrioventricular junction. Following semi-quantitative fluorescence intensity analysis of HCN4/Cx43 and validation through comparative histology, this structure was identified as the atrioventricular node (AVN) region. The AVN-to-background fluorescence intensity ratio showed no statistically significant differences among groups with increasing antibody dosage (P>0.05). The ratio increased with longer antibody perfusion times. Furthermore, no statistically significant differences in the ratio were observed among groups with extended light exposure (P>0.05). Conclusion Transaortic antegrade perfusion of fluorescently labeled antibodies can successfully image the AVN within the CCS of ex vivo rat hearts. Increasing the antibody dosage does not significantly improve the AVN imaging effect. Longer antibody perfusion time results in better imaging quality of the AVN. The fluorescent dye maintains sufficient visualization of the AVN even after prolonged (8 h) exposure to light.

Objective:To investigate the effects of macrophage(Mφ)polarization on the cementogenic differentiation of human perio-dontal ligament stem cells(hPDLSCs)and the underlying mechanism.Methods:Human monocytic THP-1 cells were induced to M0,M1 and M2 Mφ subsets,then RPM1 1640 medium or supernatants of different Mφ phenotypes were mixed with an equal volume of ce-mentoblastic induction medium to generate conditioned mediums(CMs),and termed as CM-Control,CM-M0,CM-M1 and CM-M2,respectively.hPDLSCs were cultured with different CMs,and the hPDLSCs sheets were then wrapped around treated dentin matrix(TDM)to generate cell sheet/dentin complexes.The complexes were subcutaneously implanted into nude mice.The cementum-like tissue formation was evaluated by HE staining,immunofluorescent staining(IMF)and qRT-PCR were used to detect the expression level of cementogenic differentiation-related markers bone sialoprotein(BSP),cementum attachment protein(CAP)and cementum pro-tein-1(CEMP-1),oxidant-antioxidant system-related markers superoxide dismutase 1(SOD1)and nuclear factor erythroid 2-related factor 2(NRF2),mitophagy-related markers PTEN induced putative kinase 1(PINK1)and microtubule asso ciated proteins 1A/1B light chain 3(LC3).Results:In vivo,CM-M2-treated hPDLSCs(CM-M2)group formed more cementum-like tissues and expressed higher protein levels of CAP,CEMP-1,SOD1,PINK1 and LC3 than that in other groups.In vitro tests showed that,compared with CM-Control group,hPDLSCs incubated with CM-M2 increased the levels of BSP(P＜0.01),CAP(P＜0.001),CEMP-1(P＜0.01)and SOD1(P＜0.05),while no statistically significant difference was detected for NRF2(P＞0.05),and increasedthe expression of PINK1(P＜0.05).Conclusion:M2 Mφ regulate the cementogenic differentiation of hPDLSCs possibly via modulating oxidant-antioxidant system and mitophagy.

The positive regulation of bone-forming osteoblast activity and the negative feedback regulation of osteoclastic activity are equally important in strategies to achieve successful alveolar bone regeneration. Here, a molybdenum (Mo)-containing bioactive glass ceramic scaffold with solid-strut-packed structures (Mo-scaffold) was printed, and its ability to regulate pro-osteogenic and anti-osteoclastogenic cellular responses was evaluated in vitro and in vivo. We found that extracts derived from Mo-scaffold (Mo-extracts) strongly stimulated osteogenic differentiation of bone marrow mesenchymal stem cells and inhibited differentiation of osteoclast progenitors. The identified comodulatory effect was further demonstrated to arise from Mo ions in the Mo-extract, wherein Mo ions suppressed osteoclastic differentiation by scavenging reactive oxygen species (ROS) and inhibiting mitochondrial biogenesis in osteoclasts. Consistent with the in vitro findings, the Mo-scaffold was found to significantly promote osteoblast-mediated bone formation and inhibit osteoclast-mediated bone resorption throughout the bone healing process, leading to enhanced bone regeneration. In combination with our previous finding that Mo ions participate in material-mediated immunomodulation, this study offers the new insight that Mo ions facilitate bone repair by comodulating the balance between bone formation and resorption. Our findings suggest that Mo ions are multifunctional cellular modulators that can potentially be used in biomaterial design and bone tissue engineering.
Bone Regeneration ; Cell Differentiation ; Ions/pharmacology* ; Molybdenum/pharmacology* ; Osteoclasts ; Osteogenesis ; Printing, Three-Dimensional ; Tissue Scaffolds/chemistry*

Objective    To explore the mechanism of volume-related mitral regurgitation (MR) from the anatomy of mitral valve. Methods    A total of 32 patients with ventricular septal defect (VSD) combined MR meeting inclusion criteria in West China Hospital from September 2018 to November 2019 were enrolled in this study. The direction relative to the cardiac axis: the deviation of the MR bundle along the left atrial wall was eccentric, otherwises it was central. There were 23 patients of VSD and eccentric MR (EMR, a VSD-EMR group), including 10 males and 13 females aged 21 (10, 56) months, and 9 patients of VSD and central MR (CMR, a VSD-CMR group), including 4 males and 5 females aged 26 (12, 87) months. Besides, 9 healthy children were enrolled in a control group, including 4 males and 5 females aged 49 (15, 72) months. All patients underwent transthoracic echocardiography (TTE) examination at 2 weeks before surgery and 6 months after surgery, respectively, The MR degree, end-diastolic volume (EDV), end-systolic volume (ESV), stroke volume (SV), ejection fraction (EF), antero-posterior diameter (AP), annulus circumference (AC), commissural diameter (CD) were assessed. Results    Before operation, EDV, ESV, SV, AP, AC and CD in the VSD-EMR and VSD-CMR groups were significantly larger or longer than those in the control group (P<0.05); after operation, EDV, ESV, SV, AP and CD decreased compared with those before operation (P<0.05), but there was no significant difference compared with the control group (P>0.05). Compared with the control group, AC was slightly decreased (P<0.05). There was no significant difference in EF between and within groups before and after operation (P>0.05). The improvement rate of MR was 78.9%(15/19) in the VSD-EMR group and 100.0% (9/9) in the VSD-CMR group. Conclusion    After unloading of volume, the valve structure is back to normal except AC. The improvement rate of MR in the VSD-EMR group is lower than that in the VSD-CMR group, which may indicate that the mechanism of VSD-EMR is more complicated.

Objective: To understand the incidence of stroke in the population of Jinchang Cohort and the relationship between metabolic diseases and stroke, and provide scientific evidence for the prevention and treatment of stroke in the population. Methods: The epidemiological investigation data and physical examination data of the 33 042 follow-up participants in Jinchang Cohort were collected for a prospective cohort study. Restricted cubic splines functions was used to analyze the dose-response relationship between metabolic indexes and the risk of stroke incidence. Results: 1) The incidence rate of stroke in Jinchang Cohort was 1.59%, and the standardized incidence rate was 3.99%. 2) Hypertension (male HR=2.20, female HR=4.45) and dyslipidemia (male HR=1.49, female HR=1.79) were the risk factors of stroke incidence in the population and diabetes had influence on the incidence of stroke only in the males (HR=1.79), while obesity had influence only in the females (HR=1.64). The more kinds of metabolic diseases, the higher risk of stroke incidence was. 3) Systolic blood pressure had a non-linear dose-response correlation with the risk of stroke incidence, while diastolic blood pressure had a positive linear correlation with the risk of stroke incidence. Conclusions The incidence of stroke in Jinchang Cohort population was high compared with both domestic level and oversea level. The patients with metabolic diseases were the population at high-risk for stroke, and more attention should be paid to them in the prevention and treatment of stroke. Diastolic blood pressure might be more closely related to stroke.

Objective    To investigate the reliability and safety of the technique of percutaneous left ventricular transapical access guided by cardiac three dimensional CT angiography (3D-CTA) combined with echocardiography applied in structural heart defects. Methods    The clinical data of 9 patients (7 males and 2 females with a median age of 50 years ranging from 43 to 64 years) with paravalvular leaks closed by percutaneous left ventricular transapical access in West China Hospital, from April 2015 to August 2018, were retrospectively analyzed. We applied preoperative cardiac 3D-CTA to define the puncture site and trace, which was established by combining with real-time guidance of transesophageal echocardiography (TEE/3D-TEE), and an occluder was deployed at the apical access point for hemostasis with real-time guidance of transthoracic echocardiography (TTE). Results    The puncture needles were successfully introduced into the left ventricular cavity at one time in all patients without injury of lung tissue, coronary artery or papillary muscle. There was no occluder displacement or apex bleeding. One patient developed pleural effusion caused by intercostal artery injury. Conclusion    That cardiac 3D-CTA is used to define puncture sites and trace with advantages of simplicity and repeatability. A safe access and secure exit of left ventricle can be achieved by combining with real-time guidance of echocardiography. There are acceptable technology-related complications.

Objective Transplantation of bone marrow mesenchymal stem cells (BMSCs) has a good prospect of application for cerebral infarction,but the environment and the inflammatory response to ischemia and hypoxia after cerebral infarction are not con-ducive to the survival of transplanted cells. This article investigated the effects of Shuxue Tongmai capsule(SXTM) combined with BM-SCs transplantation on the improvement of cerebral ischemic injury in rats. Methods A model of middle cerebral artery occlusion was es-tablished in Sprague-Dawley(SD) rats using thread method and these 15 SD rats were randomly divided into model group,BMSCs group and combination therapy group (BMSCs transplantation combined with SXTM treatment). At 24h after modeling,rats in combination therapy group were given tail vein injection of 1 mL BMSCs suspension (2× 109 per/L) and gavage administration of SXTM 0. 64 g/kg. Rats in BMSCs group were given tail vein injection of 1 mL BMSCs suspension (2×109 per/L) and gavage administration of equal volume of sa-line. For model group,the rats were given tail vein injection of equal volume of PBS and gavage administration of equal volume of sa-line. Neurologic function was assessed before cell transplantation and at 3,7,14,28 days after cell transplantation to check the injury of neurologic function. At 28 days after transplantion,the rats were decapitated after anesthesia to take brain tissues for immunohisto-chemical detection of vascular endothelial growth factor(VEGF) and brain-derived neurotrophic factor(BDNF) protein expression. Mor-phological changes of the brain tissue and apoptosis in cortical neurons were observed and detected by hematoxylin-eosin staining and TUNEL,respectively. Results At 7,14,28 days after transplantation,the neurological defect score in combination therapy group was significantly lower than those of model group and BMSCs group(P<0.05). In each group,the neurological defect score at 3 days after transplantation was significantly decreased compared with those before transplantation(P<0.05). In the same group,the neurologi-cal defect scores at 14,28 days after transplantation were significantly decreased compared with those at 7 days after transplantation (P<0.05). The neurological defect scores at 14,28 days after transplantation were significantly decreased compared with those at 7 days after transplantation(P<0.05). The neurological defect score at 28 day after transplantation was significantly decreased compared with that at 7 day after transplantation(P<0.05). At 28 day after transplantation,the number of apoptotic cells in combination therapy group (51.40±4.04) was significantly fewer than those of model group (74.80±5.31) and BMSCs group (67.20±4.66) and the num-ber of apoptotic cells in BMSCs group was significantly decreased compared with model group(P<0.05). The results of immunohisto-chemistry showed that the VEGF and BDNF positive cells in the cerebral ischemic region of rats were brownish or sepia in color. Com-pared with model group,the expression levels of VEGF and BDNF protein in BMSCs group and combination therapy group were signifi-cantly increased (P<0.05),and that of combination therapy group was significantly increased compared with BMSCs(P<0.05). Conclusion SXTM combined with BMSCs transplantation can promote neurological recovery from cerebral ischemia by increasing the protein expression of VEGF and BDNF and reducing neuronal apoptosis.

OBJECTIVETo evaluate the diagnostic utility of Warthin-Starry silver stain, immunohistochemistry and transmission electron microscopy in the detection of human Bartonella henselae infection and pathologic diagnosis of cat scratch disease (CSD).

METHODSThe paraffin-embedded lymph node tissues of 77 histologically-defined cases of cat scratch disease collected during the period from January, 1998 to December, 2008 were retrieved and studied using Warthin-Starry silver stain (WS stain) and mouse monoclonal antibody against Bartonella henselae (BhmAB stain). Five cases rich in bacteria were selected for transmission electron microscopy.

RESULTSUnder electron microscope, the organisms Bartonella henselae appeared polymorphic, round, elliptical, short rod or bacilliform shapes, ranged from 0.489 to 1.110 microm by 0.333 to 0.534 microm and often clustered together. Black short rod-shaped bacilli arranged in chains or clumps were demonstrated in 61.0% (47/77) of CSD by WS stain. The organisms were located outside the cells and lie mainly in the necrotic debris, especially near the nodal capsule. In 72.7% (56/77) of the cases, dot-like, granular as well as few linear positive signals were observed using BhmAB immunostain and showed similar localization. Positive results for both stains were identified in 59.7% (46/77) of the cases. When applying both stains together, Bartonella henselae was observed in 74.0% (57/77) of the case. The difference between the results obtained by WS stain and BhmAB immunostain was of statistical significance (P < 0.05).

CONCLUSIONSBartonella henselae is the causative pathogen of cat scratch disease. WS stain, BhmAB immunostain and transmission electron microscopy are helpful in confirming the histologic diagnosis. Immunostaining using BhmAB can be a better alternative than WS stain in demonstrating the organisms.

Adolescent ; Adult ; Aged ; Antibodies, Bacterial ; blood ; Bartonella henselae ; immunology ; isolation & purification ; ultrastructure ; Cat-Scratch Disease ; diagnosis ; microbiology ; pathology ; Child ; Child, Preschool ; Humans ; Immunohistochemistry ; methods ; Infant ; Lymph Nodes ; pathology ; ultrastructure ; Microscopy, Electron, Transmission ; Middle Aged ; Paraffin Embedding ; Staining and Labeling ; methods ; Young Adult

OBJECTIVETo study the roles of histologic examination and polymerase chain reaction in diagnosis of toxoplasmic lymphadenitis (TL).

METHODSForty-six archival cases of histologically diagnosed TL, encountered during the period from April, 1999 to September, 2009 and with the paraffin-embedded lymph node tissue blocks available, were enrolled into the study. The presence of genome fragments of Toxoplasma gondii (T. gondii) was analyzed using semi-nested polymerase chain reaction (PCR). Thirty cases of one or two histopathologic triad of TL as the controls.

RESULTSThe positive rate of PCR in TL group was 76.1% (35/46), as compared to 10.0% (3/30) in the control group. The difference was of statistical significance. The sensitivity and specificity of the histologic triad in diagnosing TL was 92.1% (35/38) and 71.1% (27/38), respectively. The predictive value of positive and negative PCR results was 76.1% (35/46) and 90.0% (27/30). respectively.

CONCLUSIONSThe high specificity but low sensitivity of applying the histologic triad in diagnosing TL cases may be due to the occurrence of atypical histologic pattern. The sensitivity is improved with the use of semi-nested PCR in detecting T. gondii DNA.

Adolescent ; Adult ; Aged ; Child ; DNA, Protozoan ; analysis ; Female ; Genome, Protozoan ; genetics ; Humans ; Lymph Nodes ; pathology ; Lymphadenitis ; diagnosis ; genetics ; parasitology ; pathology ; Male ; Middle Aged ; Paraffin Embedding ; Polymerase Chain Reaction ; methods ; Staining and Labeling ; Toxoplasma ; genetics ; isolation & purification ; Toxoplasmosis ; diagnosis ; genetics ; parasitology ; pathology ; Young Adult

OBJECTIVETo investigate the clinicopathologic features of lymphoplasmacytic lymphoma (LPL) with Waldenström's macroglobulinemia (WM) and to evaluate the usefulness of immunophenotype analysis in diagnosis and differential diagnosis of the tumor.

METHODSA total of 40 cases of LPL with WM diagnosed according to the 2008 World Health Organization classification of tumors of hematopoietic and lymphoid tissues were analyzed using immunophenotype and follow-up information.

RESULTSThe mostly common initial clinical presentations were non-specific symptoms, such as fatigue, anemia and hemorrhage. Lymphadenopathy, splenomegaly and hepatomegaly were found in 42.5%, 20.0% and 12.5% of the patients respectively. The pattern of bone marrow involvement included mixed type (47.2%), diffuse type (41.7%) and interstitial type (11.1%). The nodal architecture was completely destroyed in one case and partially effaced with residual germinal centers and dilated sinuses in 8 cases. All of the neoplastic cells expressed CD20 and CD79a. Neoplastic plasma cells were positive for CD138 and CD79a. No cases expressed CD5. Four cases weakly expressed CD23. No significant prognosis related factors were identified in the survival analysis.

CONCLUSIONSLPL with WM is a rare indolent small B-cell lymphoma, which is commonly seen, in older male patients. The tumor frequently involves bone marrow and shows various clinical manifestations. Combination analyses of the bone marrow biopsy histology, immunophenotypic study and clinical data, especially the serum examination are important for the diagnosis of LPL with WM.

Adult ; Aged ; Aged, 80 and over ; Antigens, CD20 ; metabolism ; Bone Marrow ; metabolism ; pathology ; CD79 Antigens ; metabolism ; Diagnosis, Differential ; Female ; Follow-Up Studies ; Humans ; Immunoglobulin M ; blood ; Immunophenotyping ; Leukemia, Lymphocytic, Chronic, B-Cell ; metabolism ; pathology ; Lymphatic Metastasis ; Lymphoma, B-Cell, Marginal Zone ; metabolism ; pathology ; Lymphoma, Follicular ; metabolism ; pathology ; Lymphoma, Mantle-Cell ; metabolism ; pathology ; Male ; Middle Aged ; Multiple Myeloma ; metabolism ; pathology ; Neoplasm Invasiveness ; Survival Rate ; Syndecan-1 ; metabolism ; Waldenstrom Macroglobulinemia ; immunology ; metabolism ; pathology