Study on the Effectiveness of Newman's System Model of Care in the Perioperative Period of Endometrial Cancer Patients.

Objective: The purpose of this study was to investigate the effectiveness of the Newman System model of care in the perioperative period for patients with endometrial cancer, to investigate the impact on psychological well-being, quality of life, and patient satisfaction. Methods: One hundred and eight patients with endometrial cancer who were treated in our hospital from January 2020 to January 2023 were selected as research subjects and were randomly divided into the control group (n=54) and the research group (n=54); the control group received conventional nursing care, and the research group added nursing care based on the conventional nursing care with Newman's systematic nursing care model including primary, secondary and tertiary care, comparing the psychological state score, quality of life score, coping style score, and satisfaction with nursing care of the two groups. Results: After the intervention, the anxiety self-assessment scale (SAS) and depression self-assessment scale (SDS) scores decreased in both groups, with the research group having lower scores than the control group (P < .05). After the intervention, all Oncology Quality of Life Core Questionnaire (QLQ-C30) scores increased in both groups, with the research group having higher scores than the control group (P < .05). After the intervention, avoidance and submission scores decreased in both groups, with the research group having lower scores than the control group (P < .05); coping scores increased in both groups, with the research group having higher scores than the control group (P < .05). Satisfaction with care was higher in the research group than in the control group (P < .05). Conclusion: The Newman's system model of care is effective in the perioperative period of endometrial cancer, helping to reduce patients' negative emotions, optimize coping, improve quality of life and nursing satisfaction, and has great value in clinical application and promotion.

Preliminary study on the mechanism by which exosomes derived from human exfoliated deciduous teeth improve the proliferation and osteogenic inhibitory effect of glucocorticoid-induced BMSCs.

BACKGROUND: Steroid-induced osteonecrosis of the femoral head (SONFH) is a disease characterized by a collapsed femoral head caused by the overuse of glucocorticoids. Dysfunction of bone marrow mesenchymal stem cells (BMSCs) is an important pathological feature of SONFH. In this study, we investigated whether exosomes from SHEDs (stem cells from human exfoliated deciduous teeth) have a therapeutic effect on glucocorticoid-induced inhibition of proliferation and osteogenesis in BMSCs, and elucidated the underlying mechanisms involved. METHODS: Primary dental pulp cells were isolated and cultured from human deciduous tooth pulp, SHEDs were isolated and purified by the limiting dilution method and exosomes were isolated from the supernatants of SHEDs by ultracentrifugation. The cell surface markers CD31, CD34, CD45, CD73, CD90 and CD105 were detected by flow cytometry. A Cell-Counting-Kit-8 assay was used to detect cell activity. ALP and Alizarin Red staining were used to identify osteogenic differentiation ability, and exosomes were identified using transmission electron microscopy, NanoFCM and Western blotting. PKH67 fluorescence was used to track the uptake of exosomes by BMSCs. Transcriptome analysis combined with quantitative real-time PCR was used to explore the underlying mechanism involved. RESULTS: Exosomes secreted by SHEDs can be endocytosed by BMSCs, and can partially reverse the inhibitory effects of glucocorticoids on the viability and osteogenic differentiation of BMSCs. Transcriptome sequencing analysis revealed that the differentially expressed mRNAs regulated by SHED-derived exosomes were enriched mainly in signaling pathways such as the apoptosis pathway, the PI3K-Akt signaling pathway, the Hippo signaling pathway and the p53 signaling pathway. qPCR showed that SHED-derived exosomes reversed the dexamethasone-induced upregulation of HGF and ITGB8 expression and the inhibition of EFNA1 expression, but further increased the dexamethasone-induced downregulation of IL7 expression. In conclusion, SHED-derived exosomes partially reversed the inhibitory effects of glucocorticoids on BMSC proliferation and osteogenesis by inhibiting the expression of HGF, ITGB8 and IL7, and upregulating the expression of EFNA1.

Molecular mechanisms involved in regulating protein activity and biological function of MST3.

Mammalian sterile 20-like (Ste20-like) protein kinase 3 (MST3) or serine/threonine-protein kinase 24 (STK24) is a serine/threonine protein kinase that belongs to the mammalian STE20-like protein kinase family. MST3 is a pleiotropic protein that plays a critical role in regulating a variety of events, including apoptosis, immune response, metabolism, hypertension, tumor progression, and development of the central nervous system. The MST3-mediated regulation is intricately related to protein activity, post-translational modification, and subcellular location. Here, we review the recent progress on the regulatory mechanisms against MST3 and its-mediated control of disease progression.

Hemophagocytic lymphohistiocytosis in two patients following treatment with pembrolizumab: two case reports and a literature review.

Background: Hemophagocytic lymphohistiocytosis (HLH) is a rare and life-threatening state of immune hyperactivation. It has the highest mortality rate among all hematological immune-related adverse events (irAEs) of immune checkpoint inhibitors (ICIs) when treating various cancers. However, the predisposing factors of HLH have rarely been mentioned in previous research. Case Description: Herein, we report 2 cases of HLH following treatment with pembrolizumab. A patient was diagnosed with thymic carcinoma (TC) and possible Sjögren's syndrome (SS), while another was diagnosed with non-small cell lung cancer (NSCLC) and Epstein-Barr virus (EBV) infection, and both were positive for antinuclear antibodies. Both cases experienced transient immune-related fever on day 7 after pembrolizumab administration and splenomegaly on day 10. Then recurrent high-grade fever appeared, and liver function impairment, highly elevated ferritin, and hypertriglyceridemia were tested. After the diagnosis of HLH, both patients were treated with dexamethasone and etoposide without relapse in our follow-up. Conclusions: Considering the widespread use of ICIs and the high mortality rate of HLH, the immune-related fever, splenomegaly, and other signs of hyperinflammation after the infusion of ICIs, are worthy of attention to the presence of HLH. Preexisting autoimmune diseases (ADs) or positive antibodies, concomitant infection, and the setting of thymic epithelial tumors (TET) may be predisposing factors for HLH. And increased caution is needed before the initiation of ICIs for patients with 2 or more predisposing factors.

Experimental and Molecular Simulation Studies of Huadian Oil Shale Kerogen.

Microscopic details on the intrinsic chemical reactivity of Huadian oil shale kerogen associated with electron properties of kerogen were investigated by the combination of experimental analyses and molecular simulations. Multimolecular structure models of kerogen with different densities were constructed for examining the accuracy of the proposed kerogen model. Results revealed that the simulated density of the kerogen model is in good agreement with the experimental value. Evaluation of the kerogen model revealed that the energy optimization process is mainly related to the change in the bond angle caused by atom displacement. According to the results from the Hirshfeld analysis of atomic charges, the S atoms in thiophene and S=O structures exhibit positive charges. By contrast, the concentration of electrons on the S atom led to the electronegativity of the sulfhydryl group. To investigate the distribution characteristics of electrons in kerogen, the molecular electrostatic potential (MEP) of a complete kerogen molecule was calculated. Notably, this is the first report of an MEP diagram of the kerogen model that can provide valuable information on the determination of electrophilic or nucleophilic reaction sites for kerogen.

CircATXN7 contributes to the progression and doxorubicin resistance of breast cancer via modulating miR-149-5p/HOXA11 pathway.

Breast cancer is a frequent tumor threatening the health of women. Circular RNAs (circRNAs) play vital roles in cancer progression and chemoresistance. Herein, we mainly investigate the role and potential mechanism of circRNA ataxin 7 (circATXN7; circ_0066436) in breast cancer. RNA expression levels were detected via quantitative real-time PCR (qPCR), western blot and immunohistochemistry. Cell viability and half inhibitory concentration (IC50) of doxorubicin were assessed by cell counting kit-8 (CCK-8) method. Cell proliferation, migration and invasion were determined by CCK-8, 5-ethynyl-2'-deoxyuridine, colony formation and transwell assays. The binding relationship between microRNA-149-5p (miR-149-5p) and circATXN7 or homeobox A11 (HOXA11) was validated via dual-luciferase reporter assay and RNA immunoprecipitation assay. Xenograft assay was conducted to analyze the effect of circATXN7 on doxorubicin resistance of breast cancer. CircATXN7 and HOXA11 levels were enhanced, whereas miR-149-5p level was declined in breast cancer tissues and cells. CircATXN7 silencing suppressed breast cancer development and doxorubicin resistance. Additionally, circATXN7 upregulated HOXA11 via absorbing miR-149-5p, thereby inducing breast cancer cell progression and reducing doxorubicin sensitivity. Besides, depletion of circATXN7 enhanced doxorubicin sensitivity in vivo. Interference of circATXN7 inhibited breast cancer progression and doxorubicin resistance via mediating miR-149-5p/HOXA11 axis, which might provide a possible biomarker for breast cancer therapy.

Enhanced Magnetic Properties of FeSiAl Soft Magnetic Composites Prepared by Utilizing PSA as Resin Insulating Layer.

Thermosetting organic resins are widely applied as insulating coatings for soft magnetic powder cores (SMPCs) because of their high electrical resistivity. However, their poor thermal stability and thermal decomposition lead to a decrease in electrical resistivity, thus limiting the annealing temperature of SMPCs. The large amount of internal stress generated by soft magnetic composites during pressing must be mitigated at high temperatures; therefore, it is especially important to find organic resins with excellent thermal stabilities. In this study, we prepared SMPCs using poly-silicon-containing arylacetylene resin, an organic resin resistant to high temperatures, as an insulating layer. With 2 wt % PSA as an insulating layer and annealed at 700 °C for 1 h, the FeSiAl SMPCs achieved the best magnetic properties, including the lowest core loss of 184 mW/cm3 (measured at 0.1 T and 50 kHz) and highest permeability of 96.

An antitumor peptide RS17-targeted CD47, design, synthesis, and antitumor activity.

BACKGROUND: CD47 is a widely expressed transmembrane protein located on the surface of somatic cells. It mediates a variety of cellular processes including apoptosis, proliferation, adhesion, and migration. An important role for CD47 is the transmission of a "Don't eat me" signal by interacting with SIRPα on the macrophage surface membrane, thereby preventing the phagocytosis of normal cells. However, cancer cells can take advantage of this autogenous signal to protect themselves from phagocytosis, thus enabling immune escape. Blocking the interaction between CD47 and SIRPα has proven to be effective in removing cancer cells. The treatment of various cancers with CD47 monoclonal antibodies has also been validated. METHODS: We designed and synthesized a peptide (RS17), which can specifically bind to CD47 and block CD47-SIRPα signaling. The affinity of RS17 for CD47-expressing tumor cells was determined, while the inhibition of CD47-SIRPα signaling was evaluated in vitro and in vivo. RESULTS: The results indicated that RS17 significantly promotes the phagocytosis of tumor cells by macrophages and had a similar therapeutic effect compared with a positive control (CD47 monoclonal antibodies). In addition, a cancer xenograft mouse model was established using CD47-expressing HepG2 cells to evaluate the effect of RS17 on tumor growth in vivo. Using ex vivo and in vivo mouse models, RS17 demonstrated a high inhibitory effect on tumor growth. CONCLUSIONS: Based on our results, RS17 may represent a novel therapeutic peptide for cancer therapy.

Quantitative kinetic parameters of primary tumor can be used to predict pelvic lymph node metastasis in early-stage cervical cancer.

PURPOSE: To investigate the role of kinetic parameters of primary tumor derived from dynamic contrast-enhanced MRI (DCE-MRI) in predicting pelvic lymph node metastasis (PLNM) in patients with cervical cancer. METHODS: 66 women with newly diagnosed cervical cancer were included between July 2017 and August 2019. All patients had a FIGO stage IB-IIA cancer and treated with hysterectomy and bilateral lymphadenectomy. Kinetic parameters of the primary tumor were derived from DCE-MRI data. The tumor diameter, ADC value, kinetic parameters, and nodal short-axis diameter were compared between patients with or without PLNM. Logistic regression analysis was used to determine the independent predictors for PLNM and receiver operator characteristic curve was used to evaluate the predictive performance. RESULTS: There were 20 patients with PLNM and 46 patients without PLNM. Tumor diameter, the efflux rate constant (Kep), and nodal short-axis diameter were significantly higher in patients with PLNM (P < 0.01). Multivariate logistic regression analysis showed that Kep and short-axis diameter were independent predictors for PLNM. Combining Kep and nodal short-axis diameter yielded the highest area under the curve (AUC) of 0.839. Combined with Kep, the sensitivity, specificity, negative predictive value, and positive predictive value of nodal short-axis diameter increased from 0.500, 0.957, 0.815, and 0.833 to 0.600, 0.978, 0.923, and 0.849, respectively. With 1.113 min-1 as threshold, the sensitivity and specificity values of Kep in predicting PLNM in patients with normal-sized lymph nodes were 0.909 and 0.667, respectively. CONCLUSIONS: Kep of primary tumor can be used as a surrogate marker to predict PLNM in cervical cancer.

Regulatory role and mechanism of the inhibition of the Mcl-1 pathway during apoptosis and polarization of H37Rv-infected macrophages.

BACKGROUND: Myeloid cell leukemia-1 (Mcl-1) plays an important role in the clearance of Mycobacterium tuberculosis (MTB) infection. It has the effect of anti-apoptosis, protecting macrophages that have engulfed pathogens and preventing pathogen clearance. Meanwhile, the MAPK signaling pathway plays a significant role in regulating Mcl-1 expression during tuberculosis infection. In the case of latent infection and active infection, the apoptosis and polarization of macrophages have a great influence during MTB infection, so we discussed the effect of Mcl-1 on apoptosis and polarization. Then, further discussed its mechanism. METHODS: An infected RAW264.7 macrophage model was established to investigate the regulatory role and mechanism of the Mcl-1 pathway inhibition during apoptosis and polarization of H37Rv infection. First, Mcl-1 protein and mRNA was identified by western blotting and Real-Time Polymerase Chain Reaction (RT-PCR). RAW264.7 macrophage apoptosis was detected by flow cytometry. RT-PCR was utilized to detect Bax, Caspase-3, Cyt-c and Bcl-2 mRNA expression. Next, Then the expression levels of inflammation factors CD86, CD206, iNOS, Fizz1, IL-6, IL-10, TNF-α, and TGF-ß was detected by ELISA. SEM was used to observe macrophages phenotype. Finally, Bax, Bcl-2 and Bcl-xl the expression was detected by western blotting. Confocal microscopy was used to analyze mitochondrial membrane potential using the JC-10 kit. RESULTS: In this study, we found that inhibiting the Mcl-1 expression signaling pathway led to infection by different virulence Mycobacterium tuberculosis, as well as changes in Mcl-1 protein and mRNA expression. Concomitantly macrophage apoptosis rate also changed, While, two phenotypic states of M1 and M2 appeared in the infected cells. We also found that the mitochondrial pathway was activated, the expression of its related genes Bax, casepase3, and Cyt-c, increased, whereas that of Bcl-2 decreased, and the mitochondrial membrane depolarization function was changed. CONCLUSIONS: We found that Mcl-1 affected the apoptosis and polarization of macrophages infected by Mycobacterium tuberculosis, mainly M1 in the early stage and M2 in the later stage. In addition, mitochondria played a crucial role in this process.

Enhanced Safety and Antitumor Efficacy of Switchable Dual Chimeric Antigen Receptor-Engineered T Cells against Solid Tumors through a Synthetic Bifunctional PD-L1-Blocking Peptide.

Chimeric antigen receptor (CAR)-engineered T cells (CAR-Ts) therapy has an excellent efficacy in cancer treatment, especially its impressive results in hematological malignancies. Unfortunately, its application on solid tumors is challenged by the off-target effects caused by lacking of tumor specific antigens and the immunosuppression caused by the tumor microenvironment. We constructed a switchable dual receptor CAR-T cell (sdCAR-T) whose activity relied upon double antigens (mesothelin and fluorescein isothiocyanate) and was strictly controlled by a "switch" (FPBM) consisting of a PD-L1 blocking peptide conjugated to fluorescein isothiocyanate. SdCAR-T cells were activated only when FPBM and cognate tumor cells expressing both PD-L1 and mesothelin coexist. Importantly, long-term proliferation experiments in vitro and the pharmacodynamic study in vivo showed a stronger antitumor activity of this system compared to the second generation mesothelin CAR-T cells. In view of this novel treatment paradigm being safer and more effective than traditional CAR-T cells, it may become a new strategy for the treatment of solid tumors.

Bioinformatics analysis of microRNA expression between patients with and without latent tuberculosis infections.

Tuberculosis (TB) is a globally prevalent infectious disease. The mechanisms of latent TB infection (LTBI) remain to be fully elucidated and may provide novel approaches for diagnosis. As therapeutic targets and molecular diagnostic markers, microRNAs (miRs) have been studied and utilized in various diseases. In the present study, the differentially expressed miRs (DEMs) in LTBI were screened and analyzed to determine the underlying mechanisms and identify potential biomarkers, thereby contributing to the diagnosis of LTBI. The GSE25435 and GSE29190 datasets from Gene Expression Omnibus were selected for analysis. The 2 datasets were analyzed individually using the Bioconductor package to screen the DEMs with specific cut-off criteria [P<0.01 and |log (fold change)|≥1]. Target gene prediction and interaction network construction were performed using Targetscan, the Search Tool for the Retrieval of Interacting Genes and Proteins and Cytoscape individually, and were merged using the latter tool. The hub genes were finally selected based on their degree of connectivity (DC). Gene Ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were performed using the KEGG and GENCLIP. A total of 144 DEMs were identified from the 2 datasets. By exploring the overlapping miRs in the two datasets, Homo sapiens (hsa)-miR-29a and hsa-miR-15b were identified to be decreased, while hsa-miR-576-5p, hsa-miR-500 and hsa-miR-155 were identified to be upregulated. hsa-miR-500a-3p and hsa-miR-29a-3p, as well as 4 genes, namely cell division cycle (CDC)42, actin α1, skeletal muscle (ACTA1), phosphatase and tensin homolog (PTEN) and fos proto-oncogene (FOS), were selected as the key factors in this regulatory network. A total of 9 signaling pathways, including phosphoinositide-3 kinase (PI3K)/AKT and 11 biological processes, were identified to be associated with LTBI. In conclusion, the present analysis identified hsa-miR-500a-3p and hsa-miR-29a-3p, as well as CDC42, ACTA1, PTEN and FOS, as the most promising biomarkers and therapeutic candidates for LTBI. The PI3K/AKT signaling pathway is the key signaling pathway implicated in LTBI, and an in-depth investigation of the efficiency of PI3K/AKT signaling inhibitors may be used to prevent a chronic state of infection in LTBI.

Effect of gap junctions on RAW264.7 macrophages infected with H37Rv.

BACKGROUND: Apoptosis and inflammation have been shown to play an important role in the mechanisms involved in the pathogenesis of Mycobacterium tuberculosis (MTB) infection. When macrophages undergo apoptosis and polarization, gap junctions (GJs) may be needed to provide conditions for their functions. Connexin 43 (Cx43) and connexin 37 (Cx37) are the main connexins in macrophages that participate in the formation of GJ channels. METHODS: An H37Rv infection RAW264.7 macrophage model was established to investigate the associate between connexins and host macrophage immune defense response after MTB infection. First, Real-time Polymerase Chian Reaction (RT-PCR) was used to detect the mRNA expression of Cx43 and Cx37. Cx43 protein expression and location was detected by western blotting and immunofluorescence. Confocal microscope was used to assay the gap junctional intercellular communication (GJIC). Then, electron microscope used to observe the morphology of macrophages. Finally, RAW264.7 macrophage apoptosis and mitochondrial membrane potential was detected by flow cytometry, and the expression of inflammation factors such as CD86, CD206, and IL-6, IL-10, TNF-α, and TGF-ß were detected by Real-time PCR and enzyme-linked-immunosorbent serologic assay (ELISA). RESULTS: H37Rv infection significantly promoted host macrophage Cx43 mRNA and protein expression (increased 1.6-fold and 0.3-fold respectively), and enhanced host macrophage GJIC. When host macrophage cell-to-cell communication induced by H37Rv infection, the apoptosis rate and inflammatory factors expression also increased. CONCLUSIONS: The results confirm that H37Rv infection can obviously induce host macrophage Cx43 expression and enhance GJIC, which may implicated in host macrophage inflammatory reaction, to regulate the release of inflammatory factors and/or initiate apoptosis to activate host immune defense response.

Biological and Biomechanical Evaluation of Autologous Tendon Combined with Ligament Advanced Reinforcement System Artificial Ligament in a Rabbit Model of Anterior Cruciate Ligament Reconstruction.

OBJECTIVE: To compare the biomechanical and histological changes in a rabbit model after reconstructing the anterior cruciate ligament (ACL) with solely autologous tendon and with autologous tendon combined with the ligament advanced reinforcement system (LARS) artificial ligament. METHODS: Anterior cruciate ligament reconstruction was performed in 72 knees from 36 healthy New Zealand white rabbits (bodyweight, 2500-3000 g). The Achilles tendons were harvested bilaterally. The left ACL were reconstructed solely with autografts (autologous tendon group), while the right ACL were reconstructed with autografts combined with LARS ligaments (combined ligaments group). The gross observation, histological determination, and the tension failure loads in both groups were evaluated at 12 weeks (n = 18) and 24 weeks (n = 18) postoperatively. RESULTS: Gross examination of the knee joints showed that all combined ligaments were obviously covered by a connective tissue layer at 12 weeks, and were completely covered at 24 weeks. Fibrous tissue ingrowth was observed between fascicles and individual fibers in the bone-artificial ligament interface at both time points; this fibrovascular tissue layer localized at the bone-artificial ligament interface tended to be denser in specimens obtained at 24 weeks compared with those obtained at 12 weeks. The tension failure loads of the knees were similar in the autologous tendon group and the combined ligaments group at 12 weeks (144.15 ± 3.92 N vs. 140.88 ± 2.75 N; P > 0.05), and at 24 weeks (184.15 ± 1.96 N vs. 180.88 ± 3.21 N; P > 0.05). CONCLUSION: Reconstructing the ACL in rabbits using autologous tendon combined with the LARS artificial ligament results in satisfactory biointegration, with no obvious immunological rejection between the autologous tendon and the artificial ligament, and is, therefore, a promising ACL reconstruction method.

Double-bundle anatomical medial patellofemoral ligament reconstruction with lateral retinaculum plasty can lead to good outcomes in patients with patellar dislocation.

PURPOSE: To evaluate the clinical outcomes of double-bundle anatomical medial patellofemoral ligament (MPFL) reconstruction combined with lateral retinaculum plasty versus isolated MPFL reconstruction for patellar dislocation. METHODS: From 2011 to 2013, 64 patients underwent either double-bundle anatomical MPFL reconstruction combined with lateral retinaculum plasty (Group A, 32 patients), or isolated MPFL reconstruction (Group B, 32 patients). Clinical evaluations were performed 2 years post-operatively and included determination of the number of patients with patellar redislocation, the patellar tilt angle, lateral patellar shift, subjective symptoms, and functional outcomes as evaluated with the Kujala score and Lysholm score. RESULTS: No dislocation or subluxation occurred during the 25-month follow-up. Radiographically, the patellar tilt angle was 6.0° ± 1.7° in Group A and 6.7° ± 2.0° in Group B, without a significant difference (n. s). The lateral patellar shift was 8.0 ± 2.6 mm in Group A and 8.5 ± 2.4 mm in Group B (n. s). The mean Kujala score was 91.8 ± 3.7 in Group A and 91.5 ± 3.6 in Group B post-operatively (n. s), and the mean Lysholm score was 92.5 ± 3.8 and 90.8 ± 4.9, respectively (n. s). CONCLUSIONS: Double-bundle anatomical MPFL reconstruction with lateral retinaculum plasty is a promising procedure that provides a new option for patellar dislocation. LEVEL OF EVIDENCE: III.

CT-based morphological analysis of the posterior femoral condyle in patients with trochlear dysplasia.

BACKGROUND: The anterior part of the distal femur in trochlear dysplasia has been well investigated; however, to date, posterior morphological characteristics have not been well studied. This study aimed to evaluate whether the posterior femoral condyle in patients with trochlear dysplasia differs from those without trochlear dysplasia. METHODS: Computed tomography scans of 75 knees with trochlear dysplasia and 55 knees with normal anatomy of the patellofemoral joint were analyzed retrospectively. Three observers assessed the width, length, and height of the posterior condyle between the two groups. The intra-class correlation coefficient was used to evaluate inter-observer reliability. The independent Student's t-test was used to assess the statistical significance of the qualitative variables. RESULTS: There was excellent inter-observer reliability (intra-class correlation coefficient 0.91-0.99) for all of the quantitative measurements. There were significant differences between trochlear dysplastic and normal knees. The trochlear dysplasia group had a larger medial posterior condyle and smaller lateral posterior condyle than the control group. Furthermore, proportion of the posterior condyle in the distal femur markedly differed between the two groups: in the trochlear dysplasia group, the medial posterior condyle accounted for a bigger proportion, while the lateral posterior condyle accounted for a smaller proportion. CONCLUSION: Patients with trochlear dysplasia have different posterior femoral condyles compared with those without trochlear dysplasia. Patients with this condition have bigger medial posterior condyles and smaller lateral posterior condyles. A greater amount of attention needs to be paid to this abnormality.

Mcl-1 signals pathway inhibitors in mouse peritoneal macrophage apoptosis infected with the Xinjiang strain of M. tuberculosis.

A present, myeloid cell leukemia-1 (Mcl-1) was suggested as a potential new target for controlling latent TB infection. Therefore, we investigated the role of the Mcl-1 signalling pathway in mouse peritoneal macrophages infected with XJ-MTB, aiming at finding a new strategy for TB management in Xinjiang. We using TUNEL, Immunohistochemical analysis, ELISA, HE, RT-PCR and Western blot detected macrophages apoptosis, the damage of mice tissues and the expression of apoptosis genes and proteins. Results found that inhibition of the Mcl-1 signalling pathway not only reduced the survival of intracellular XJ-MTB, but also increased peritoneal macrophage apoptosis in latent XJ-MTB-infected mouse peritoneal macrophages and relieved the pathological damage of mouse organs infected with XJ-MTB, especially MAPK signalling pathway inhibitor PD98059 (P<0.05). Moreover, after inhibitor PD98059 treated mouse peritoneal macrophages infected with XJ-MTB, Bcl-2, Bax and Mcl-1 were reduced, while Cytochrome-c and Caspase-8 protein levels were significantly increased, and Cytochrome-c protein levels was significant higher than Caspase-8 (P<0.05). In conclusion, the MAPK signalling pathway inhibitor PD98059 down-regulated Mcl-1 expression and effectively increased macrophage apoptosis in mice infected with XJ-MTB. Furthermore, it also relief pathological organ damage and promote the elimination of inflammation. The intrinsic apoptotic pathway plays a predominant role in the regulatory role.

The regulatory role of Mcl-1 in apoptosis of mouse peritoneal macrophage infected with M. tuberculosis strains that differ in virulence.

PURPOSE: Myeloid cell leukaemia-1 (Mcl-1) is a valuable target in tumour treatments. However, several reports have suggested that Mcl-1 may play a role in tuberculosis infection. Therefore, we investigated the function of Mcl-1 in tuberculosis infection and the underlying regulatory mechanism. METHODS: Mcl-1-shRNA was used to down-regulate Mcl-1 expression in BCG-, H37Ra-, H37Rv- and XJ-MTB-infected mouse peritoneal macrophages. TUNEL staining detected macrophage apoptosis. The colony-forming units (CFUs) were determined to assess the Mycobacterium tuberculosis (MTB) clearance after down-regulating Mcl-1. Immunohistochemical analysis of Mcl-1 expression in mouse peritoneal macrophages was performed. Haematoxylin and eosin staining detected pathologic damage of the liver, spleen, lung, and kidney in mice. Real-time PCR and Western blotting determined the expression of cytochrome-c in Mcl-1-shRNA-treated mouse peritoneal macrophages infected with MTB strains that differ in virulence. RESULTS: Mcl-1-shRNA significantly promoted host macrophage apoptosis and cytochrome-c induction, and the apoptotic induction of the XJ-MTB and H37Rv strains was stronger than the H37Ra and BCG strains (P<0.05). Apoptotic protein cytochrome-c levels continued to increase in mouse peritoneal macrophages infected MTB before and after treatment, Caspase-8 levels only slightly increased after Mcl-1-shRNA-treated (P<0.05), but the increase of Cytochrome-c have no significant differences compared with Caspase-8 levels (P>0.05). CONCLUSION: Mcl-1-shRNA intervention effectively down-regulated Mcl-1 expression, significantly increased host macrophage apoptosis, and induced cytochrome-c expression in mouse peritoneal macrophages infected with MTB strains of different virulence, and these changes were influenced by the virulence of the MTB strains. The mitochondria-mediated intrinsic apoptotic pathway play an important role before Mcl-1-shRNA-trated, and then with the extrinsic apoptotic pathway co-regulate host macrophage apoptosis.

A small hairpin RNA targeting myeloid cell leukemia-1 enhances apoptosis in host macrophages infected with Mycobacterium tuberculosis.

Myeloid cell leukemia-1 (Mcl-1) plays an important role in various cell survival pathways. Some studies indicated that the expression of Mcl-1 was upregulated in host cells during infection with the virulent Mycobacterium tuberculosis strain, H37Rv. The present study was designed to investigate the effect of inhibiting Mcl-1 expression both in vivo and in vitro on apoptosis of host macrophages infected with M. tuberculosis using a small hairpin (sh)RNA. Mcl-1 expression was detected by the real time-polymerase chain reaction, western blotting, and immunohistochemistry. Flow cytometry and transmission electron microscopy were used to measure host macrophage apoptosis. We found elevated Mcl-1 levels in host macrophages infected with M. tuberculosis H37Rv. The expression of Mcl-1 was downregulated efficiently in H37Rv-infected host macrophages using shRNA. Knockdown of Mcl-1 enhanced the extent of apoptosis in H37Rv-infected host macrophages significantly. The increased apoptosis correlated with a decrease in M. tuberculosis colony forming units recovered from H37Rv-infected cells that were treated with Mcl-1-shRNA. Reducing Mcl-1 accumulation by shRNA also reduced accumulation of the anti-apoptotic gene, Bcl-2, and increased expression of the pro-apoptotic gene, Bax, in H37Rv-infected host macrophages. Our results showed that specific knockdown of Mcl-1 expression increased apoptosis of host macrophages significantly and decreased the intracellular survival of a virulent strain of M. tuberculosis. These data indicate that interference with Mcl-1 expression may provide a new avenue for tuberculosis therapy.

Suppression of Mcl-1 induces apoptosis in mouse peritoneal macrophages infected with Mycobacterium tuberculosis.

The effect of myeloid cell leukemia-1 (Mcl-1) inhibition on apoptosis of peritoneal macrophages in mice infected with Mycobacterium tuberculosis was investigated and the primary signaling pathway associated with the transcriptional regulation of Mcl-1 was identified. Real-time PCR and western blotting indicated that Mcl-1 transcript and protein expression are upregulated during infection with virulent M. tuberculosis H37Rv and Xinjiang strains but not with attenuated M. tuberculosis strain H37Ra or Bacillus Calmette-Guérin. Mcl-1 transcript and protein expression were downregulated by specific inhibitors of the Janus kinase/signal transducer and activator of transcription (JAK/STAT), mitogen-activated protein kinase (MAPK) and phosphoinositol 3-kinase (PI3K) pathways (AG490, PD98059 and LY294002, respectively). The strongest inhibitor of Mcl-1 expression was PD98059, the MAPK inhibitor. Flow cytometry demonstrated that the rate of apoptosis in peritoneal macrophages is significantly higher in mice infected with M. tuberculosis and the rate of apoptosis is correlated with the virulence of the strain of M. tuberculosis. Apoptosis was found to be upregulated by AG490, PD98059 and LY294002, whereas inhibition of the MAPK pathway sensitized the infected macrophages to apoptosis. Taken together, these results suggest that specific downregulation of Mcl-1 significantly increases apoptosis of peritoneal macrophages and that the MAPK signaling pathway is the primary mediator of Mcl-1 expression.