Background Gastric cancer (GC) is an aggressive malignant tumor with a high degree of heterogeneity, and its immune microenvironment is closely associated with tumor growth, development and drug resistance. Therefore, a classification system of gastric cancer based explicitly on the immune microenvironment context might enrich the strategy for gastric cancer prognosis and therapy. Methods A total of 668 GC patients were collected from TCGA-STAD (n = 350), GSE15459 (n = 192), GSE57303 (n = 70) and GSE34942 (n = 56) datasets. Three immune-related subtypes (immunity-H, -M, and -L) were identified by hierarchical cluster analysis based on the ssGSEA score of 29 immune microenvironment-related gene sets. The immune microenvironment-related prognosis signature (IMPS) was constructed via univariate Cox regression, Lasso-Cox regression and multivariate Cox regression, and nomogram model combining IMPS and clinical variables was further constructed by the “rms” package. RT-PCR was applied to validate the expression of 7 IMPS genes between two human GC cell lines (AGS and MKN45) and one normal gastric epithelial cell line (GES-1). Results The patients classified as immunity-H subtype exhibited highly expressed immune checkpoint and HLA-related genes, with enriched naïve B cells, M1 macrophages and CD8 T cells. We further constructed and validated a 7-gene (CTLA4, CLDN6, EMB, GPR15, ENTPD2, VWF and AKR1B1) prognosis signature, termed as IMPS. The patients with higher IMPS expression were more likely to be associated with higher pathology grade, more advanced TNM stages, higher T and N stage, and higher ratio of death. In addition, the prediction values of the combined nomogram in predicting 1-year (AUC = 0.750), 3-year (AUC = 0.764) and 5-year (AUC = 0.802) OS was higher than IMPS and individual clinical characteristics. Conclusions The IMPS is a novel prognosis signature associated with the immune microenvironment and clinical characteristics. The IMPS and the combined nomogram model provide a relatively reliable predictive index for predicting the survival outcomes of gastric cancer.

Background:Nanoparticle albumin-bound paclitaxel (nab-paclitaxel) is an optimized and improved derivative of paclitaxel with superior efficacy and fewer adverse reactions, and it is widely used in the treatment of advanced gastric cancer. However, there is a paucity of data regarding the safety and efficacy of nab-paclitaxel combined with oxaliplatin (LBP) and tegafur in the treatment of patients with advanced gastric cancer.Methods:This analysis is a prospective, single-center, open-label, historically controlled real-world study designed to include 10 patients with advanced gastric cancer treated with nab-paclitaxel combined with LBP and tegafur gimeracil oteracil potassium. The primary and main efficacy outcomes are safety indicators, including the incidence of adverse drug reactions and adverse events (AEs), as well as the outliers of laboratory indicators and vital signs. The secondary efficacy outcomes are overall survival (OS), objective response rate (ORR), disease control rate (DCR), and proportion of dose suspensions, dose reductions and discontinuations.Discussion:Based on the findings of previous studies, we wished to assess the safety and efficacy of nab-paclitaxel combined with LBP and tegafur in the treatment of advanced gastric cancer. The trial requires constant contact and monitoring. The purpose is to determine a superior protocol in terms of patient survival, and pathological and objective response.Trial Registration:This trial has been registered with the Clinical Trial Registry: NCT05052931 (registration date: 2021/9/12).

Background Breast cancer (BC) is the most common malignant tumor in women and widely known for its poor prognosis. More and more research has discovered that cyclin E1 (CCNE1) plays an important role in progression of various types of cancer. But its specific mechanism in BC progression still needs further research to explore. Methods At first, we determined the expression and prognostic value of CCNE1 through The Cancer Genome Atlas (TCGA) database and The Genotype-Tissue Expression (GTEx) data. Then, we predicted the upstream non-coding RNAs of CCNE1 through StarBase, GEPIA, and Kaplan-Meier plotter database. We further studied the correlation of CCNE1 expression with BC immune cell infiltration, biomarkers of immune cells and immune checkpoints expression through TIMER and GEPIA databases. Results The results suggested that CCNE1 was significantly upregulated in BC and its high expression was correlated with poor prognosis in BC patients. Next, we identified long noncoding RNA (lncRNA) LINC00511 / microRNA-195-5p (miR-195-5p) / CCNE1 axis as the most potential pathway that could regulate CCNE1 expression in BC through StarBase, GEPIA, and Kaplan-Meier plotter database. Furthermore, our in-depth research discovered that CCNE1 expression level was significantly correlated with tumor immune cell infiltration, biomarkers of immune cells, and immune checkpoint expression in BC. conclusions In summary, high expression level of CCNE1 was significantly correlated with poor prognosis, tumor immune infiltration and escape in BC.

BACKGROUND:This study aims to retain normal knee kinematics after knee replacement surgeries by reconstructing anterior cruciate ligament during total knee arthroplasty.METHOD:We use computational simulation tools to establish four dynamic knee models, including normal knee model, posterior cruciate ligament retaining knee model, posterior cruciate ligament substituting knee model, and anterior cruciate ligament reconstructing knee model. Our proposed method utilizes magnetic resonance images to reconstruct solid bones and attachments of ligaments, and assemble femoral and tibial components according representative literatures and operational specifications. Dynamic data of axial tibial rotation and femoral translation from full-extension to 135 were measured for analyzing the motion of knee models.FINDINGS:The computational simulation results show that comparing with the posterior cruciate ligament retained knee model and the posterior cruciate ligament substituted knee model, reconstructing anterior cruciate ligament improves the posterior movement of the lateral condyle, medial condyle and tibial internal rotation through a full range of flexion. The maximum posterior translations of the lateral condyle, medial condyle and tibial internal rotation of the anterior cruciate ligament reconstructed knee are 15.3 mm, 4.6 mm and 20.6 at 135 of flexion.INTERPRETATION:Reconstructing anterior cruciate ligament in total knee arthroplasty has been approved to be an more efficient way of maintaining normal knee kinematics comparing to posterior cruciate ligament retained and posterior cruciate ligament substituted total knee arthroplasty.

Background: A series of epidemiological studies have attempted to evaluate the impact of 309T>G polymorphism in MDM2 gene frequently identified as a susceptibility loci for various cancers on malignant sarcomas, however the reported conclusions remain inconsistent and elusive. We pooled all usable data sets in order to systematically assess the association between 309T>G polymorphism and sarcoma risk. Methods: To identify as many informative studies with complete data as possible, we searched a number of databases (PubMed, EBSCO, BIOSIS, the Cochrane Library, ISI Web of Science, Wiley Online Library and Embase). Inclusion criteria were defined to select the eligible studies. The fixed effects meta-analysis was properly used to calculate the pooled ORs and 95% CIs. Major findings: We eventually identified six studies evaluating the association of sarcoma risk with 309T>G polymorphism. People with 309-GG were found to have 43% greater risk of sarcoma relative to people with 309-TT (OR, 1.43; 95% CI, 1.01 +/- 2.03; Pheterogeneity, 0.45). In the G vs. T genetic model, the risk reduced to 19% (OR, 1.19; 95% CI, 1.01 +/- 1.40; Pheterogeneity, 0.50). Statistical data showed no significant heterogeneity or publication bias in the meta-analysis. Conclusion: These data demonstrate that 309T>G polymorphism located within the MDM2 gene may act as modifier factor for sarcomas. A weakness of this analysis is that the findings cannot be explainable when the subtypes are separated and additional larger investigations are needed to identify the role of 309T>G polymorphism in each form of sarcoma.

A T>G single nucleotide polymorphism (SNP, rs2279744) of the MDM2 gene has been investigated in sarcoma community, but the findings are conflicting. This study was designed to well define the relationship between SNP rs2279744 and sarcoma risk. We did a systematic computerized search of the PubMed, Web of Science, and Science Direct databases to identify the human case–control studies investigating the relationship between SNP rs2279744 and sarcoma risk with complete genetic data. Pooled odds ratios (ORs) were calculated with the Mantel–Haenszel fixed-effect model or the DerSimonian and Laird random effects model to estimate the risk of sarcoma. Overall analysis included five independent studies. On the whole, the T/G genotype or the combined G/G and T/G genotypes appeared to be associated with approximately 1.40-fold higher risk of sarcoma relative to the T/T genotype (T/G vs. T/T: OR 1.33, 95 % CI 1.00–1.77; G/G + T/G vs. T/T: OR 1.42, 95 % CI 1.08–1.85). We noted that the Caucasian populations showed a similarly increased risk of sarcoma ascribed to the carriage of the same genotypes (T/G vs. T/T: OR 1.41, 95 % CI 1.05–1.90; G/G + T/G vs. T/T: OR 1.49, 95 % CI 1.13–1.97). This meta-analysis provides evidence that MDM2 SNP rs2279744 may be significantly associated with increased risk of sarcoma in Caucasian individuals.

Objective To investigate the inhibitory effects of curcumin (Cur) on proliferation and apopotosis of human osteosarcoma cell line MG-63 cells.Methods The cultured MG-63 cells were treated with Cur (12.5,25.0,50.0,100.0 μmol/L) for 12 h.The morphologic changes of MG-63 cells were observed by inverted microscope.The survival rate of MG-63 cells was measured by using methyl thiazol tetrazolium (MTT) assay.Cell counting of kit-8 (CCK-8) assay was used to test the growth inhibitory rate.Annexin V/propidium iodide (PI) staining was applied to detect the apoptosis.The survival ratios and the adhesion capacity of MG-63 were detected except the lactate dehydrogenase (LDH) releasing.Results Different concentrations of Cur (12.5,25.0,50.0,100.0 μmol/L) observed in this study have significant effects on the survival ratio of MG-63.The Cur could significantly inhibit the growth of MG-63 in a dose and time dependent manner and induce G2/M Phase cell cycle arrest.After 12 h,cell inhibition rate of 47.23％,24 h after the cell was 65.23％.Cur 12.5,25.0,50.0,100.0 μmol/L group for 24 h,MG-63 cell apoptosis rate was (11.24 ± 0.07)％,(28.23 ± 0.35)％,(50.21 ± 1.71)％ and (41.22 ±1.85) ％,50.0 μmol/L curcumin could induce apoptosis obviously.Conclusion When Cur increased from 12.5 μmol/L to 25.0 μmol/L,its MG-63 cells at 24h growth inhibition rate (13.56 ± 1.88) ％ to (48.27 ±2.32)％,50.0 μmol/L group 24 h growth inhibition rate to 65.23％.50.0 μmol/L of Cur on the MG-63 cells apoptosis induced by the most obvious,for 24 h,apoptosis rate was (50.21 ± 1.71)％.

A T>G single nucleotide polymorphism (SNP, rs2279744) of the MDM2 gene has been investigated in sarcoma community, but the findings are conflicting. This study was designed to well define the relationship between SNP rs2279744 and sarcoma risk. We did a systematic computerized search of the PubMed, Web of Science, and Science Direct databases to identify the human case–control studies investigating the relationship between SNP rs2279744 and sarcoma risk with complete genetic data. Pooled odds ratios (ORs) were calculated with the Mantel–Haenszel fixed-effect model or the DerSimonian and Laird random effects model to estimate the risk of sarcoma. Overall analysis included five independent studies. On the whole, the T/G genotype or the combined G/G and T/G genotypes appeared to be associated with approximately 1.40-fold higher risk of sarcoma relative to the T/T genotype (T/G vs. T/T: OR 1.33, 95 % CI 1.00–1.77; G/G + T/G vs. T/T: OR 1.42, 95 % CI 1.08–1.85). We noted that the Caucasian populations showed a similarly increased risk of sarcoma ascribed to the carriage of the same genotypes (T/G vs. T/T: OR 1.41, 95 % CI 1.05–1.90; G/G + T/G vs. T/T: OR 1.49, 95 % CI 1.13–1.97). This meta-analysis provides evidence that MDM2 SNP rs2279744 may be significantly associated with increased risk of sarcoma in Caucasian individuals.

We aimed to evaluate the expression of microRNA-182 (miR-182) in triple-negative breast cancer (TNBC) tissues and the TNBC cell line MDA-MB-231 and to investigate the effects of mirR-182 on the cellular behavior of MDA-MB-231 and the expression of the target gene profilin 1 (PFN1), thus providing new methods and new strategies for the treatment of TNBC. Quantitative real-time PCR (qRT-PCR) was utilized to evaluate the expression of miR-182 in TNBC tissues, relatively normal tissues adjacent to TNBC and the TNBC cell line MDA-MB-231. Forty-eight hours after the MDA-MB-231 cells were transfected with the miR-182 inhibitor, qRT-PCR was utilized to detect the changes in miR-182 expression levels, and an MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay was utilized to determine the effects of miR-182 on cell viability. Flow cytometry was adopted to determine whether miR-182 affects the proliferation rates and apoptosis levels of the MDA-MB-231 cells. The transwell migration assay method was used to investigate the effects of miR-182 on the migration of the MDA-MB-231 cells. A luciferase reporter gene system was applied to validate that PFN1 was the target gene of miR-182. Western blot was used to measure the effects of miR-182 on the PFN1 protein expression levels in the cells. qRT-PCR results showed that compared with the relatively normal tissues adjacent to TNBC, miR-182 expression was significantly increased in the TNBC tissues and the MDA-MB-231 cells (p<0.01). Compared with the control group, MDA-MB-231 cells transfected with the miR-182 inhibitor and incubated for 48 h showed significantly decreased miR-182 expression (p<0.01). The results of an MTT assay showed that inhibition of miR-182 in MDA-MB-231 cells led to significantly reduced cell viability (p<0.05). Flow cytometry analysis indicated that inhibition of miR-182 expression resulted in significantly decreased cell proliferation (p<0.05) and significantly increased levels of apoptosis (p<0.05). The results of a transwell migration assay showed that after inhibited of miR-182 expression, the number of cells passing through the transwell membranes was significantly decreased (p<0.05). The results from a luciferase reporter gene system showed that compared with the control group, the relative luciferase activity of the group transfected with the miR-182 inhibitor was significantly increased (p<0.05). Western blot analysis showed that compared with the control group, PFN1 protein expression levels were significantly increased in the MDA-MB-231 cells transfected with the miR-182 inhibitor and incubated for 48 h (p<0.05). In conclusion, miR-182 is upregulated in TNBC tissues and cells. It promotes the proliferation and invasion of MDA-MB-231 cells and could negatively regulate PFN1 protein expression. Treatment strategies utilizing inhibition of miR-182 expression or overexpression of the PFN1 gene might benefit patients with TNBC.

Some recent,large-scale, forward-looking studies have ascertained that enhanced blood sugarreducing treatments can effectively decrease likelihood of long-term complications of jiabetes and lengthen prospected life spans of diabetic patients. With glycemic control reaching targets,risks of hypoglycaemia increase.Hypoglycaemia severely hinders good control of blood sugar and serious low blood sugar levels lead to death of diabetes. Thus treatment of diabetes not only need make glycemic control meet targets but also need reduce occurrence of hypoglycaemia. This paper addresses causes of hypoglycaemia in Type 2 diabetes therapy, its clinical symptoms and types as well as workable solutions.

Although more than 40 beta-defensins have been identified in rat epididymis, little is known about their regulation or their relation to male infertility caused by inflammation. Using a rat model of epididymitis induced by lipopolysaccharide (LPS), we examined expression of SPAG11E (also known as Bin1b), a caput epididymis-specific beta-defensin in rat. Unlike the expression of other beta-defensins in various epithelial cells with upregulated expression after LPS stimulation, expression of SPAG11E was significantly decreased by LPS at the mRNA and protein levels. LPS treatment also significantly decreased both sperm binding to SPAG11E and sperm motility, and supplementation of the spermatozoa with recombinant SPAG11E in vitro remarkably increased both SPAG11E binding and motility of sperm. To clarify whether decreased expression is a common pattern of epididymal beta-defensins after LPS stimulation, we examined the expression of another 12 epididymal beta-defensins expressed in the caput epididymis. For nine of these beta-defensins, expression was decreased, but for the other three, expression remained unaffected. These findings demonstrate that LPS-induced epididymitis can decrease the expression of epididymal beta-defensins and that disruption of SPAG11E expression is involved in the impairment of sperm motility.

Few studies have stressed on the sensitivity of stress distribution in different mechanical properties of the articular cartilage and subchondral bone. The purpose of this study was to establish parametric variations of mechanical factors individually and examine how these biomechanical effects influenced the cartilage and subchondral bone plate stress fields in the hip joint. A finite element model including acetabulum and proximal femur was established to study the stress change associated with the thinning of cartilage, the increasing of subchondral bone modulus and the thickening of subchondral bone plate individually. The stress distributions in bone/cartilage interface were evaluated. Sensitivity of the stress magnitudes to the parametric changes was also analyzed. The results indicated that cartilage thinning has more significant effect than subchondral bone modulus increasing or thickening on the shear stress levels in subchondral bone/cartilage interface. Subchondral bone plate modulus increasing has mild effect on the shear stress in subchondral bone/cartilage interface. Cartilage thinning acts as a major influence on the development of the articular cartilage damage.

Objective:To investigate the regulatory effect of dexamethasone on NF-κB and its target gene cyclinD1 in human prostate cancer cell line PC-3,so as to explore the anti-proliferation mechanism of dexamethasone.Methods: The inhibitory effects of dexamethasone on growth of PC-3 cells were determined by MTT.The basic transcriptional activity of NF-κB in many kinds of tumor cells and the effects of dexamethasone on the activation of PC-3 cell NF-κB were determined by reporter gene assay.The expression of cyclinD1 protein was determined by Western blots.Results: Dexamethasone inhibited the growth of PC-3 cells and NF-κB was constitutively activated in the PC-3 cells.Dexamethasone also inhibited the activation of NF-κB and down-regulated the expression of cyclinD1 protein in PC-3 cells. Conclusion: Dexamethasone can inhibit cell growth of prostate cancer cell line PC-3,which might be related to the inhibitory effect of dexamethasone on the transcriptional activation of NF-κB and cyclinD1 protein.

Objective: To investigate the role of CD44v3 in adhesion and invasion behavior of human lung carcinoma cell A549 induced by hyaluronan in vitro. Methods: The RNA interference plasmid expressing shRNA (short hairpin RNA) against CD44v3 was designed and constructed, and was used for transfection of the A549 cell line. The expression of CD44v3 before and after transfection of the A549 cell line was examined by the reverse transcription polymerase chain reaction (RT-PCR) and western blot methods. Both Adhesive and invasive ability before and after transfection of the A549 cell line were determined by the plate adhesion model and Boyden chamber model. Results: Compared to pre-transfection, the A549 cell CD44v3 mRNA and expression of proteins transfected with RNA interference plasmid as well as the cell population sticking to the flat plate and penetrating septum of Boyden chamber decreased significantly (P<0.01). Conclusions: CD44v3 mediates the in vitro adhesion and invasive behavior of hyaluronic acid-induced human lung carcinoma cell line A549.

OBJECTIVE:To observe the effect of dexamethasone (Dex) on the proliferation of human ovarian cancer cells of the line HO-8910, and explore the role of RhoB signaling pathway in this process.METHODS:Human ovarian cancer cells of the line HO-8910he were cultured in culture fluids with or without different concentrations of Dex. The cell growth levels in anchor-dependent and anchor-independent manner were detected by MTT and soft agar assay. Another HO-8910 cells were inoculated in gel with different concentrations of Dex. HO-8910 was transfected with the eukaryotic expression plasmid RhoB-wt, blank plasmids pcDNA3 and RhoB-RNAi, and then the mRNA expression of RhoB, a small GTPase gene, was examined by semi-quantitative RT-PCR. and the protein expressions of RhoB, p-Akt, and p21(cip1/waf1) and p27, both cyclin kinase inhibitors (CDIs), were detected by Western blotting. HO-8910 cells were co-transfected with the reporter gene p21-luc containing p21 promoter and marker reporter gene pRL-tk-luc, then treated with Dex for 24 h. Western blotting was used to detect the transcription of p21(cip1/waf1) gene.RESULTS:The RhoB mRNA expression was significantly increased 2 hours after the treatment of 100 nM Dex, and peaked 4 hours later as high as 2.5 times that of the control group. Western blotting showed that the RhoB protein expression increased along the increase of the Des concentration. The protein expression of RhoB in the HO-8910 cells transfected with RhoB-wt was 2.02 times that in the HO-8910 cells transfected with blank plasmid, and the protein expression of RhoB in the HO-8910 cells transfected with RhoB-RNAi was 36% of that of the blank plasmid group (P < 0.01). The HO-8910 cell proliferation of the RhoB-RNA1 group was not significantly different from that of the control group, however, the proliferation of the HO-8910 cell treated by 100 nM Dex for 6 days was significantly inhibited with an inhibition rate of 13% (P < 0.01). Western blotting showed that Dex down-regulated the p-Akt protein expression. Dex time and dose-dependently up-regulated the protein expression of p21(cip1/waf1) and p27. The HO-8910 cells co-transfected with p21-luc and pRL-tk-luc and then treated with Dex for 24 h showed an higher p21-luc activity, 1.72 times that of the control group (P < 0.05).CONCLUSION:The mechanism of inhibiting the proliferation by Dex in ovarian cancer cells may involve the depression of PI3K/p-Akt, and then up-regulation of RhoB and its downstream signal molecules p21(cip1/waf1) and p27 proteins.

A simple method was advanced to wipe starch off RNA and obtain purer total RNA. This method was successfully applied in Northern blot and RNA-array experiments to study the expression of starch branching enzyme gene (Rbe1) and Wx gene, which were important for starch synthesis. And the results indicate that on the whole, the expression of Wx gene was linear with amylose content of rice endosperm, and there were two Rbe1 homogenes in rice endosperm, the longer expressed only in endosperm and the shorter in all tissues.

The recently discovered aging-dependent large accumulation of point mutations in the human fibroblast mtDNA control region raised the question of their occurrence in postmitotic tissues. In the present work, analysis of biopsied or autopsied human skeletal muscle revealed the absence or only minimal presence of those mutations. By contrast, surprisingly, most of 26 individuals 53 to 92 years old, without a known history of neuromuscular disease, exhibited at mtDNA replication control sites in muscle an accumulation of two new point mutations, i.e., A189G and T408A, which were absent or marginally present in 19 individuals younger than 34 years. These two mutations were not found in fibroblasts from 22 subjects 64 to 101 years of age (T408A), or were present only in three subjects in very low amounts (A189G). Furthermore, in several older individuals exhibiting an accumulation in muscle of one or both of these mutations, they were nearly absent in other tissues, whereas the most frequent fibroblast-specific mutation (T414G) was present in skin, but not in muscle. Among eight additional individuals exhibiting partial denervation of their biopsied muscle, four subjects >80 years old had accumulated the two muscle-specific point mutations, which were, conversely, present at only very low levels in four subjects ≤40 years old. The striking tissue specificity of the muscle mtDNA mutations detected here and their mapping at critical sites for mtDNA replication strongly point to the involvement of a specific mutagenic machinery and to the functional relevance of these mutations.