Morphological Characteristics of Various Cells in Esophageal Squamous Dysplasia: Extremely Wide Morphological Spectrum.

The WHO classification of esophageal tumors divides esophageal squamous intraepithelial dysplasia into high and low grades, but does not specify its morphological spectrum. Here, the morphological characteristics of various cells were investigated in esophageal squamous (high-grade) dysplasia, and a morphological spectrum and terminology for this lesion were proposed to avoid misdiagnosis. The clinicopathological data of 540 patients with esophageal squamous dysplasia were analyzed retrospectively. According to the unique cytomorphological characteristics of the lesions and the predominant cell type, the esophageal squamous dysplasia was divided into the following morphological groups: classic type (34.6%, 187/540), basaloid subtype (10.7%, 58/540), spindle-cell subtype (4.6%, 25/540), differentiated subtype (48.9%, 264/540), and verrucous subtype (1.1%, 6/540). Gender, age, and lesions location did not differ among the subtypes (P > 0.05), while Paris classification and lesions diameter significantly differed among the subtypes (P < 0.01). Classic-type cells showed severe atypia. In the basaloid subtype, the cells were small, and resembled basal cells; most of these lesions were of the 0-IIb type with small lesion diameter. In the spindle-cell subtype, the cells and nuclei were spindle-shaped or long and spindle-shaped and arranged in parallel. Differentiated-subtype showed well-to-moderately differentiated cells, and epithelial basal cells were mature. Verrucous-subtype showed well-differentiated cells, and were characterized by verrucous or papillary structures. Esophageal squamous dysplasia has extremely wide morphological spectrum. Awareness of the spectrum of morphological presentations of this lesion, specifically the basaloid subtype, spindle-cell subtype, differentiated subtype, and verrucous subtype, is important for accurate diagnosis.

Long non-coding RNA MALAT1 promotes cell proliferation, migration and invasion by targeting miR-590-3p in osteosarcoma.

Osteosarcoma (OS) is a common malignant bone cancer and commonly occurs in adolescents and children. Long non-coding RNAs (lncRNAs) play major roles in cancer cell proliferation and metastasis. The present study aimed to investigate the potential molecular mechanism of lncRNA MALAT1 in OS. The levels of lncRNA MALAT1 and microRNA-590-3p were detected by reverse transcription-quantitative PCR in OS tissues and cells. Cell Counting Kit-8 and flow cytometry assays were conducted to assess cell proliferation and apoptosis. Cell migration and invasion were examined by Transwell assay. The levels of E-cadherin, N-Cadherin, Vimentin and Snail were measured by western blotting. The target of MALAT1 was predicted using online software and confirmed by luciferase reporter, RNA immunoprecipitation and RNA pull-down assays. The results indicated that MALAT1 was highly expressed in OS tissues and cell lines. MALAT1 knockdown promoted apoptosis and suppressed proliferation, migration, invasion and epithelial- mesenchymal transition (EMT) of OS cells. Overexpression of miR-590-3p increased cell apoptosis and hampered cell proliferation, migration, invasion and EMT in OS cells. In addition, MALAT1 knockdown upregulated the expression of miR-590-3p in OS cells. In conclusion, MALAT1 was demonstrated to suppress cell apoptosis and induce cell proliferation, migration, invasion and EMT by inhibiting miR-590-3p in OS, which indicated that MALAT1 has potential value in the diagnosis and treatment of OS.

Shear strength deterioration effect of rock mass joint surface under cyclic shear load.

Understanding the shear strength degradation mechanism of a rock mass joint surface under cyclic shear load and determining a corresponding analytical model is an important foundation for accurately evaluating the safety of rock mass engineering under seismic loads. It is worth noting that, to date, there has been a dearth of studies on the strength characteristics of joint surfaces that consider the number of loading cycles, normal load, and initial undulant angle of the structural plane. In this study, focused on the behaviour of sandstone, the particle flow code (PFC) modelling framework was used to simulate a joint surface cyclic shear test considering first- and second-order undulations. Based on the experimental results, the comprehensive effects of the number of cyclic shear cycles, the normal stress, first- and second-order undulation and the dilatancy angle on shear stress during cyclic shear were analysed. Formulas for the joint surface shear basic friction angle and dilatancy angle under cyclic shear were proposed, and a method for calculating the joint surface peak shear strength under cyclic shear considering the deterioration of the dilatancy angle and basic friction angle was established. The peak shear strength of a sample after five cycles of shearing was calculated using the proposed formula and compared with the results of numerical simulations, the Barton method, and the Homand method. The results showed that the calculated values have good consistency with the results of the numerical simulations, demonstrating the effectiveness and accuracy of the proposed formula. However, under a low normal stress, there could be errors in estimating the cyclic shear strength of the joint surface.

Shear characteristics and shear strength model of rock mass structural planes.

Accurately determining the shear strength of structural planes is crucial for evaluating the stability of rock masses. The shear test using the sawtooth structural plane usually captures the main influencing factors of its shear characteristics. In this study, the two-dimensional particle flow code (PFC2D) numerical simulation method was used to conduct shear tests on the sawtooth structural planes of rock masses with undulant angles of 10°, 20°, and 30°, respectively. With the increase in normal stress and the undulant angle, the shear failure of the structural planes was found to no longer be pure slip failure or shear failure but accompanied by a compression-induced fracture phenomenon. Based on the analysis of the shear test results, a peak shear strength model considering different undulant angles and normal stresses was proposed, and the hyperbolic function post-peak shear strength model was improved. The peak shear strength obtained from the physical direct shear tests was compared with those calculated using the proposed model, Parton model, and Shen model. The calculation error under low and high normal stress of the proposed method was found to be within an acceptable range. Additionally, when calculating the peak shear strength of a structural plane under high normal stress, applying the calculation method proposed in this study is a better option than applying the other models. Furthermore, although the variation trend of the post-peak shear strength was similar to that of the experimental results, the values obtained using the hyperbolic variation model were too large. The variation trend of the post-peak shear strength obtained using the improved function was essentially consistent with the experimental results, and the calculated values were close to the experimental results. The systematic research on the shear strength calculation model of rock mass structural planes contributes to the theoretical research of rock mass mechanics, and this study can act as a guide for landslide prediction and control projects.

Method of calculating shear strength of rock mass joint surface considering cyclic shear degradation.

When a rock mass shears along a joint surface, the shear resistance is affected by joint surface undulations and friction between the contact regions. During an earthquake, the seismic load causes dynamic deterioration of the joint surface mechanical properties, mostly reflected as follows. (1) The peak shear strength of the joint surface decreases with an increase in the shear rate. (2) Under a seismic load cyclic shear, the undulant angle αk decreases. (3) Under a dynamic load, the friction coefficient of the joint surface is reduced. By studying the cyclic shear test of the joint surface, the strength deterioration effect of the joint surface under cyclic shearing is first analysed, and the equations of the dilatation angle and the basic friction angle of the joint surface under the cyclic shearing load are proposed. Then, starting with the effect of cyclic shear deterioration on the joint surface in the rock mass and the reduction in the dynamic friction coefficient between sliding rock blocks caused by relative velocity, an equation for calculating the shear strength of a rock mass joint surface under cyclic shear loading is recommended. Through two case calculations, the shear strength obtained using the proposed method is compared with the experimental results. The results show that the model proposed in this study is in good agreement with the experimental results and can also be used to calculate the structural surface shear strength of the asperity-rich sample. However, when the calculation equation is used to estimate the cyclic shear strength of the joint surface where the sum of the initial undulation angle and the basic friction angle is greater than 70°, there may be some errors in the calculation results.

Deep learning for automatic diagnosis of gastric dysplasia using whole-slide histopathology images in endoscopic specimens.

BACKGROUND: Distinguishing gastric epithelial regeneration change from dysplasia and histopathological diagnosis of dysplasia is subject to interobserver disagreement in endoscopic specimens. In this study, we developed a method to distinguish gastric epithelial regeneration change from dysplasia and further subclassify dysplasia. Meanwhile, optimized the cross-hospital diagnosis using domain adaption (DA). METHODS: 897 whole slide images (WSIs) of endoscopic specimens from two hospitals were divided into training, internal validation, and external validation cohorts. We developed a deep learning (DL) with DA (DLDA) model to classify gastric dysplasia and epithelial regeneration change into three categories: negative for dysplasia (NFD), low-grade dysplasia (LGD), and high-grade dysplasia (HGD)/intramucosal invasion neoplasia (IMN). The diagnosis based on the DLDA model was compared to 12 pathologists using 100 gastric biopsy cases. RESULTS: In the internal validation cohort, the diagnostic performance measured by the macro-averaged area under the receiver operating characteristic curve (AUC) was 0.97. In the independent external validation cohort, our DLDA models increased macro-averaged AUC from 0.67 to 0.82. In terms of the NFD and HGD cases, our model's diagnostic sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were significantly higher than junior and senior pathologists. Our model's diagnostic sensitivity, NPV, was higher than specialist pathologists. CONCLUSIONS: We demonstrated that our DLDA model could distinguish gastric epithelial regeneration change from dysplasia and further subclassify dysplasia in endoscopic specimens. Meanwhile, achieved significant improvement of diagnosis cross-hospital.

Observation of Efficacy of Internet-Based Chronic Disease Management Model Combined with Modified Therapy of Bushenyiliu Decoction in Treating Patients with Type 2 Diabetes Mellitus and Prostate Cancer and Its Effect on Disease Control Rate.

OBJECTIVE: To explore the efficacy of Internet-based chronic disease management model combined with the modified therapy of Bushenyiliu decoction in treating patients with type 2 diabetes mellitus (T2DM) and prostate cancer and its effect on disease control rate (DCR). METHODS: 120 patients with T2DM and prostate cancer admitted to the Affiliated Hospital of Yangzhou University, Yangzhou First People's Hospital, from February 2019 to February 2020, were retrospectively analyzed and equally divided into the experimental group and the control group according to their admission order. Conventional treatment combined with the modified therapy of Bushenyiliu decoction was performed on all patients for 3 months, and the Internet-based chronic disease management model was adopted for patients in the experimental group additionally, so as to compare their short-term effect, survival time, disease progression, blood glucose indicators, immune function indicators, and type 2 Diabetes Self-Care Scale (2-DSCS) scores. RESULTS: Compared with the control group, the experimental group obtained significantly higher DCR and objective remission rate (ORR) (P < 0.05), higher survival time and disease progression (P < 0.001), better blood glucose indicators and immune function indicators (P < 0.001), and higher 2-DSCS scores (P < 0.001) after treatment. CONCLUSION: Combining the Internet-based chronic disease management model with the modified therapy of Bushenyiliu decoction can effectively enhance the self-care ability of patients with T2DM and prostate cancer, improve their blood glucose level, promote their body immunity, and comprehensively optimize the cancer control effect, which should be promoted in practice.

Development of bispecific anti-c-Met/PD-1 diabodies for the treatment of solid tumors and the effect of c-Met binding affinity on efficacy.

Although the blockade of the programmed cell death protein 1/programmed cell death ligand 1 (PD-1/PD-L1) pathway has become a promising treatment strategy for several types of cancers, the constitutive activation of c-Met in tumors may cause a low overall response rate to PD-1 inhibitors. Increasing evidence indicates that the dual inhibition of c-Met and PD-1 could improve the efficacy of anti-PD-1/PD-L1 monoclonal antibodies for tumor immunotherapy. In this study, we developed two bispecific single-chain diabodies targeting c-Met and PD-1 for the treatment of solid tumors based on protein homology modeling, and we identified that the binding affinity of diabody-mp to c-Met was 50-folds higher than that of diabody-pm. The results of in vitro studies revealed that both diabodies suppressed HGF-induced proliferation, migration, and invasion of tumor cells, inhibiting the activation of c-Met signaling by antagonizing HGF binding to c-Met. Moreover, they promoted T cell activation by blocking the PD-1 pathway, mediating tumor cellular cytotoxicity through T cell engagement. In vivo studies with mice models demonstrated that diabody-mp exhibited higher therapeutic efficacy than other structural antibodies, greatly enhancing the survival of c-Met-positive tumor-bearing mice compared to single or combined c-Met and PD-1 blockade therapy. Furthermore, diabody-mp, which had a higher c-Met binding affinity, showed better anti-tumoral activity than diabody-pm, which had a lower c-Met binding affinity. In conclusion, bispecific anti-PD-1/c-Met diabody-mp, with high c-Met-associated affinity, inhibited tumor growth by activating T cells, suggesting its therapeutic potential for c-Met-positive solid tumors.

Dual-function chimeric antigen receptor T cells targeting c-Met and PD-1 exhibit potent anti-tumor efficacy in solid tumors.

Purpose Programmed cell death 1 (PD-1), which is upregulated under the continuous induction of the tumor microenvironment, causes chimeric antigen receptor (CAR)-T cell hypofunction via interaction with programmed death ligand 1 (PD-L1). This study aimed to construct CAR-T cells that are resistant to PD-1 inhibition to improve the effect of CAR-T cells in solid tumors. Methods We constructed a type of dual-function CAR-T cell that targets tumor-associated antigen c-Met and blocks the binding of PD-1 with PD-L1. The expression of c-Met, PD-L1, and inhibitory receptors was measured using flow cytometry. The cytotoxicity, cytokine release, and differentiation level of CAR-T cells were determined using lactate dehydrogenase release assay, enzyme-linked immunosorbent assay, and flow cytometry, respectively. The levels of p-Akt, p-MAPK, caspase-3, and Bcl2 were detected by western blot. The in vivo anti-tumor effect was evaluated using tumor xenograft models. Results Dual-function CAR-T cells could mediate enhanced active signals upon encountering target antigens and had targeted cytotoxicity to target cells. However, the cytotoxicity of c-Met-CAR-PD-1+ T cells was impaired due to the interaction of PD-1 with PD-L1. By blocking the binding of PD-1 and PD-L1, the novel dual-function CAR-PD-1+ T cells could maintain cytotoxicity to PD-L1+ tumor cells. In tumor tissue, the dual-function CAR-T cells showed lower inhibitory receptor expression and lower differentiation characteristics, which resulted in potent anti-tumor effects and prolonged survival in PD-L1+ tumor xenograft models compared to single-target CAR-T cells. Conclusion These results confirm that the novel dual-function CAR-T cells exhibit stronger anti-tumor activity against solid tumors than traditional single-target CAR-T cells and present a new approach that enhance the activity of CAR-T cells in solid tumors.

A novel tetravalent bispecific antibody targeting programmed death 1 and tyrosine-protein kinase Met for treatment of gastric cancer.

Background Redirecting T cells to tumor cells using bispecific antibodies (BsAbs) is emerging as a potent cancer therapy. The main concept of this strategy is to cross-link tumor cells and T cells by simultaneously binding to cell surface tumor-associated antigen (TAA) and the CD3ƹ chain. However, immune checkpoint programmed cell death ligand-1 (PD-L1) on tumor cells or other myeloid cells upreglulated remarkablely after the treatment of CD3-binding BsAbs, leads to the generation of suppressed microenvironment for immune evasion and tumor progression. Although this resistance could be partially reversed by anti-PD-L1 treatment, targeting two pathways through one antibody-based molecule may provide a strategic advantage over the combination of BsAbs and immune checkpoint inhibitors. Methods We developed two novel BsAbs PD-1/c-Met DVD-Ig and IgG-scFv both targeting PD-1 to restore the immune effector function of T cells and engaging them to tumor cells via binding to cellular-mesenchymal to epithelial transition factor (c-Met). Binding activities, T cell activation and proliferation were analyzed by flow cytometry. Cell Cytotoxicity and cytokine release were measured using LDH release assay and ELISA, respectively. Anti-tumor response in vivo was evaluated by generate xenograft models in NOD-SCID mice. Results These bispecific antibodies exhibited effective antitumor activity against high- and low- c-Met-expressing gastric cancer cell lines in vitro and mediated strong tumor growth inhibition in human gastric cancer xenograft models. Conclusion The engagement of the PD-1/PD-L1 blockade to c-Met-overexpressing cancer cells is a promising strategy for the treatment of gastric cancer and potentially other malignancies.

The degree of overlap between the follicular dendritic cell meshwork and tumor cells in mantle cell lymphoma is associated with prognosis.

This study concerning mantle cell lymphoma (MCL) investigated retrospectively an association between patient prognosis and the percentage of the total number of lymphoma cells found in the follicular dendritic cell (FDC) meshwork, that is, the degree of overlap of lymphoma cells. Two hundred and nine MCL patients were apportioned to grades I-III, in which the CD21-positive FDC meshwork covered ≤50%, 51%-89%, and ≥90% of the tumor area, respectively. Significant differences among the grades (all, P < 0.01) were found in the following: duration of disease (from onset of clinical manifestation to diagnosis); clinical staging; extranodal involvement (non-lymphoid organs); histological subtype; and Ki-67 proliferation index (PI). After removing the aggressive variants, the overall survival rates of grade I (n = 92) and II (n = 57) patients were similar. The overall survival rates of grade III (n = 46) patients differed from that of grade I + II patients (P < 0.01). The grades negatively correlated with the Ki-67 PI value (r = -0.234, P = 0.001). At each grade the OSR of patients with Ki-67 PI ≤30% was similar to that of patients with Ki-67 >30%. In the Ki-67 PI ≤30% group, the OSRs of the patients differed significantly among the grades. In the Ki-67 >30% group the OSRs of the grades were similar. The results of multivariate Cox regression analysis showed that the degree of overlap, age and Ki-67 PI was the independent prognostic factors of the OSRs of MCL patients. Our data suggests that MCL patients in whom there was a high degree of overlap between the FDC meshwork and tumor area have a better clinical prognosis. The degree of overlap correlates well with the Ki-67 PI, which can be used to predict the prognosis of patients.

Generation and Characterization of a Bispecific Antibody Targeting Both PD-1 and c-MET.

BACKGROUND: Bispecific antibodies, BsAbs, are molecules with the ability to bind to two different epitopes on the same or different antigens. c-MET, cellular-mesenchymal to epithelial transition factor, is deregulated in many types of human malignancies. Abnormal c-MET activation in cancer correlates with poor prognosis. PD-1, programmed death-1, is an additional inhibitory receptor expressed by T cells. Blocking the interactions between PD-1 and PD-L1 has emerged as a promising immunotherapy for treating cancer. OBJECTIVES: The goal of this study was to identify a novel bispecific antibody targeting both c-MET and PD-1 as an anti-cancer therapeutic candidate. METHODS: The BsAb was produced using 293E expression system and purified by Protein A affinity chromatography. Then the binding specificity and affinity of the BsAb was examined by FACS and biolayer light interferometry. The ability of the BsAb to inhibit the proliferation of tuman cells was measured using the CellTiter 96 Aqueous One Solution Cell Proliferation Assay kit; the potential signaling pathway involved was identified by Western Blot. Cytokine secreted by PHA-L stimulated PBMC was measured by ELISA. Effects of BsAb on PBMC-mediated lysis of MKN45 cells was measured by LDH cytotoxicity assay. RESULTS: Based on the original sequences of PD-1 and c-MET mAb, a BsAb gene was designed, cloned into pCEP4 vector for expression in 293E cells. The BsAb was obtained after purification of the cell culture supernatant. It can bind to PD-1 and c-MET simultaneously, the calculated affinity was 11.5 nM for PD-1 and 9.09 nM for c-MET. The BsAb enhanced IFN-γ production over control IgG by 2-3 folds. It also inhibit the c-MET pathway activation and the proliferation of tumor cells significantly, comparable to JnJ-38877605. The BsAb showed dose-dependent cytotoxic activity against MKN45 cells. CONCLUSION: Our results indicated that a novel BsAb recognizing PD-1 and c-MET was successfully generated. It could redirect T cells to kill tumor cells, while retaining its inherent ability to restore T cells and inhibit tumor cells. With this potential, this BsAb could be developed as a therapeutic candidate for the treatment of various solid tumors.

Primary hemosiderotic fibrolipomatous tumor in bone: a case report and review of the literature.

Background and aim: Hemosiderotic fibrolipomatous tumor (HFLT) is a locally invasive tumor composed of mature adipocytes accompanied by spindle cells containing hemosiderin deposition. In 2013, it was categorized by WHO as a soft tissue tumor with uncertain differentiation. So far, the literature has reported 60 cases but primary HFLT in bone has never before been reported. We set out to investigate the clinicopathological features of primary HFLT in bone. Methods: We retrospectively reviewed the clinical, imaging, histological, and immunophenotypic features and treatment of 1 case of primary HFLT in bone, and combined this with literature discussion. Results: HFLT occurred in the lateral femoral condyle of a 50-year-old male patient, which might have been overlooked were it not for the knee-joint pain and dysfunction. CT and MRI showed osteolytic bone destruction with a clear 4.0-cm diameter boundary, diagnosed as cystic damage of the lateral condyle of the left femur. SPECT metabolism was not active. Histologically, the lesion was composed of different proportions of mature fat cells, spindle cells, and hemosiderin. Immunohistochemistry revealed spindle cells expressing vimentin, p63, but not CD34, calponin, and others. The tumor tissue was thoroughly removed by curetting, and a bone graft was carried out after immersion in anhydrous ethanol. At the 11-month postoperative follow-up, the patient was recovering well. Conclusions: Primary HFLT in bone is extremely rare. In imaging, it can easily be misdiagnosed as a bone cyst. Histological morphology of the current case is similar to that of soft tissue HFLT.

A novel bispecific c-MET/PD-1 antibody with therapeutic potential in solid cancer.

The bispecific antibody is a novel antibody, which can target two different antigens and mediate specific killing effects by selectively redirecting effector cells to the target cells. Here, we designed and synthesized a bispecific antibody (BsAb) that can bind cellular-mesenchymal to epithelial transition factor (c-MET, overexpressed in several human solid tumor), and programmed death-1 (PD-1, involved in cancer cell immune evasion) with high affinity and specificity. We found that BsAb can induce the degradation of c-MET protein in cancer cells, including MKN45, a gastric cancer cell line, and A549, a lung cancer cell line. BsAb inhibited hepatocyte growth factor (HGF)-mediated proliferation, migration, and antiapoptosis, and downregulated HGF-stimulated phosphorylation of c-MET, protein kinase B (AKT), and extracellular signal-regulated kinase (ERK1/2). BsAb can also rescue T cell activation. Furthermore, xenograft analysis revealed that BsAb markedly inhibits the growth of subcutaneously implanted tumors and chronic inflammation. On the basis of these results, we have identified a potential bispecific drug, which can effectively target c-MET and PD-1 for the treatment of human solid cancers.

Bromodomain-containing protein 2 induces insulin resistance via the mTOR/Akt signaling pathway and an inflammatory response in adipose tissue.

Insulin resistance is a major metabolic abnormality in a large majority of patients with type II diabetes. Bromodomain-containing protein 2 (Brd2), a transcriptional co-activator/co-repressor with switch mating type/sucrose non-fermenting (SWI/SNF)-like functions that regulates chromatin, suppresses adipocyte differentiation and regulates pancreatic ß-cell biology. However, the effects of Brd2 on insulin resistance remain unknown. Here, overexpression of Brd2 in white adipose tissue of wild-type (WT) mice led to insulin resistance. Brd2 overexpression induced the expression of nuclear Factor-κΒ (NF-κΒ) target genes, mainly involving proinflammatory and chemotactic factors, in adipocytes. Furthermore, it decreased the expression of DEP domain containing mTOR-interacting protein (Deptor) to enhance mechanistic target of rapamycin (mTOR) signaling, thus blocking insulin signaling. Collectively, these results provided evidence for a novel role of Brd2 in chronic inflammation and insulin resistance, suggesting its potential in improving insulin resistance and treating metabolic disorders.

Characterization and high-yield production of non-N-glycosylated recombinant human BCMA-Fc in Pichia pastoris.

B-cell maturation antigen (BCMA) fused at the C-terminus to the Fc portion of human IgG1 (BCMA-Fc) blocks B-cell activating factor (BAFF) and proliferation-inducing ligand (APRIL)-mediated B-cell activation, leading to immune disorders. The fusion protein has been cloned and produced by several engineering cell lines. To reduce cost and enhance production, we attempted to express recombinant human BCMA-Fc (rhBCMA-Fc) in Pichia pastoris under the control of the AOX1 methanol-inducible promoter. To produce the target protein with uniform molecular weight and reduced immunogenicity, we mutated two predicted N-linked glycosylation sites. The secretory yield was improved by codon optimization of the target gene sequence. After fed-batch fermentation under optimized conditions, the highest yield (207 mg/L) of rhBCMA-Fc was obtained with high productivity (3.45 mg/L/h). The purified functional rhBCMA-Fc possessed high-binding affinity to APRIL and dose-dependent inhibition of APRIL-induced proliferative activity in vitro through three-step purification. Thus, this yeast-derived expression method could be a low-cost and effective alternative to the production of rhBCMA-Fc in mammalian cell lines.

Association of Cytotoxic T-lymphocyte Antigen-4 Polymorphisms with Malignant Bone Tumor Risk: A Meta-analysis.

BACKGROUND: Previous studies have assessed the association between the Cytotoxic T-lymphocyte Antigen- 4(CTLA-4) polymorphism with the risk of malignant bone tumor, but the conclusions were inconsistent. We aimed to clarify association of cytotoxic T-lymphocyte antigen-4 polymorphisms with malignant bone tumors risk by performing a meta-analysis. MATERIALS AND METHODS: The databases including PubMed, EMBase databases and the Cochrane Library were searched to identify the eligible studies prior to January 30 2016. Odds ratio (OR) with 95% confidence interval (95%CI) were used to estimate the strengths of the association between the CTLA-4 polymorphism and the malignant bone tumor risks. The meta-analysis was performed by STATA 12.0. RESULTS: Four individual studies with a total of 1003 cases with malignant bone tumors and 1162 controls were included in our meta-analysis. The results of meta-analysis on those data demonstrated that CTLA-4 +49G>A polymorphism was associated with the risk of Ewing's sarcoma and osteosarcoma strongly (A vs. G: : OR=1.36, 95%CI:1.20-1.54, p=0.000; AA+AG vs. GG: OR=1.35, 95%CI:1.14-1.61, p=0.001; AA vs. GG: OR=2.24, 95%CI:1.67-2.99, p=0.000; AA vs. AG+GG: OR=2.00, 95%CI:1.53-2.62, p=0.000), but CTLA-4 -318C/T polymorphism was not associated with the risk of malignant bone tumor (C vs. T: OR=0.76, 95%CI:0.76-1.08, p= 0.262; CC+CT vs. TT: OR=0.70, 95%CI:0.41-1.20, p=0.198; CC vs. TT: OR=0.69, 95%CI:0.40-1.19, p= 0.183; CC vs. CT+TT: OR=0.92, 95%CI:0.75-1.13, p= 0.419). Subgroup analysis showed that there are significantly positive correlations between CTLA-4 +49G>A polymorphism and increased risks of malignant bone tumors in large size of sample (A vs. G: OR=1.347, 95%CI: 1.172,1.548, p=0.000; AA vs. GG: OR=2.228, 95%CI: 1.608,3.085, p=0.000), Ewing's Sarcoma or Osteosarcoma (A vs. G: OR=1.361, 95%CI: 1.201,1.540, p=0.000; AA vs. GG: OR=2.236, 95%CI: 1.674,2.986, p=0.000), and PCR-RFLP or Sequencing(A vs. G: OR=1.361, 95%CI: 1.201,1.540, p=0.000; AA vs. GG: OR=2.236, 95%CI: 1.674,2.986, p=0.000), but CTLA-4 -318C/T polymorphism was not associated with the risk of malignant bone tumors in diagnosis, genotype method, and sample size (all p>0.05). CONCLUSIONS: CTLA-4 +49A/G variant was associated with an increased risk of developing the malignant bone tumors, such as Ewing's sarcoma and osteosarcoma. However, it failed to show any association between CTLA-4 -318C/T polymorphism and the risk of malignant bone tumors. Future large-scale studies remain to be done to confirm our conclusions.

A rabbit model with acute thrombo-embolic pulmonary hypertension created with echocardiography guidance.

A rabbit model of acute thrombo-embolic pulmonary hypertension was developed by infusing self-thrombi into the right femoral vein and simultaneously measuring the pulmonary artery pressure via a right heart catheter and echocardiography. The model was used to explore the usefulness of an ultrasound-guided protocol. In the present study, acute thrombo-embolic pulmonary hypertension was produced in 25 of 30 healthy New Zealand rabbits; the success rate was 83%. A significant and positive correlation between the right ventricle-right atrial pressure gradient, an estimate of the pulmonary artery systolic pressure derived from tricuspid regurgitation and the pulmonary artery systolic pressure measured using the right heart catheter (r=0.765, P=0.002) was noted. During the process of establishing a rabbit model of acute thrombo-embolic pulmonary hypertension, it was demonstrated that echocardiography can be used to guide the right heart catheter to obtain pulmonary artery systolic pressure measurements, to quantify the tricuspid regurgitation jet to assess the pulmonary artery systolic pressure and to observe cardiac morphologic changes so as to evaluate cardiac function. Based on the present study, it is clear that echocardiography is valuable in improving the success rate of producing the animal model of acute thrombo-embolic pulmonary hypertension. This could ultimately facilitate preclinical research and clinical research in humans.