TLR4-MyD88-NF-κB signaling imbalances Th17 and Treg cells in thymoma with myasthenia gravis.

OBJECTIVE: Thymus is an immune organ in which pathological changes may cause autoimmune diseases, including myasthenia gravis (MG). Recent studies have focused on Toll-like receptor 4 (TLR4) signaling as the cause of such changes. In our previous study, an imbalance of T helper 17 (Th17) cells and T regulatory (Treg) cells was found in MG thymoma. These results suggest the involvement of TLR4 in the pathogenesis of thymoma MG via an alteration of the Th17/Treg balance. Here, we aimed to assess whether the TLR4-MyD88-NF-κB pathway is upregulated in MG thymoma and its relationship with Th17/Treg cells. PATIENTS AND METHODS: We collect thymoma samples from 54 patients with or without MG, detecting the expression level of TLR4, MyD88, and NF-κB in thymoma tissues. Next, we established an in vitro experiment of coculturing thymoma cells with CD4+ T cells and detected the differentiation of Th17 cells and Treg cells and their marker protein, retinoid-related orphan receptor gamma t (RORγt) and forkhead transcription factor 3 (Foxp3). RESULTS: We found TLR4, MyD88, and NF-κB expressed more in MG thymoma compared with simple thymoma. After the transwell coculturing, we observed an imbalance of Th17/Treg cells after TLR4 stimulation. CONCLUSIONS: TLR4 is stimulated in thymoma, causing an increase of Th17 cells and a decrease of Treg cells, namely an imbalance of Th17/Treg cells, resulting in MG.

Association between TP53 polymorphisms and chronic lymphocytic leukemia.

OBJECTIVE: The aim of this study was to explore the association between TP53 gene polymorphisms (rs8068934 A>G and rs218698 C>T) and chronic lymphocytic leukemia (CLL). PATIENTS AND METHODS: CLL patients who received treatment in our hospital were enrolled in this study as the disease group. Meanwhile, healthy subjects were taken as the control group. Peripheral blood samples were collected to detect TP53 gene polymorphisms at rs8068934 and rs218698, and the haplotype analysis was performed. The expression of TP53 was detected via reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Furthermore, the survival conditions were analyzed. RESULTS: The allele distribution at rs8068934 (p=0.046) and rs218698 (p=0.028) of TP53 gene was different between control group and disease group. A allele frequency at rs8068934 and T allele frequency at rs218698 were significantly higher in disease group (p<0.05). The genotype distribution at rs218698 of TP53 gene in disease group was also different from that in control group (p=0.038). The results demonstrated that CC genotype frequency in disease group was significantly lower than that in control group (p<0.05). Besides, the distribution of dominant model at rs8068934 (p=0.042) and recessive model at rs218698 (p=0.033) in disease group exhibited remarkable differences from control group, in which AA+AG frequency (dominant model) at rs8068934 and CC+CT frequency (recessive model) at rs218698 in disease group were significantly higher. Meanwhile, the distribution of AT (p=0.029) and GC (p=0.007) haplotypes at rs8068934 and rs218698 in disease group was evidently different from that in control group. The results indicated that disease group showed significantly higher frequency of AT haplotype and lower frequency of GC haplotype (p<0.05). Moreover, TP53 gene polymorphisms at rs8068934 were significantly associated with the levels of white blood cells (WBC) (p=0.000) and platelets (PLT) (p=0.035). Patients with GG genotype had significantly higher level of WBC, while those with AG genotype showed significantly lower level of PLT (p<0.05). TP53 gene polymorphisms at rs218698 were associated with the level of red blood cells (RBC) (p=0.000). Patients with CT genotype had a remarkably lower level of RBC (p<0.05). There were significant correlations of TP53 gene polymorphisms at rs8068934 (p=0.000) and rs218698 (p=0.000) with the expression of TP53. The expression of TP53 was lower in people with AA genotype at rs8068934 but higher in people with TT genotype at rs218698 (p<0.05). Furthermore, TP53 gene polymorphisms at rs8068934 (p=0.000) and rs218698 (p=0.000) were markedly associated with patients' survival. CONCLUSIONS: TP53 polymorphisms are significantly correlated with the occurrence and progression of CLL.

MiR-22-3p regulates the proliferation and invasion of Wilms' tumor cells by targeting AKT3.

OBJECTIVE: In this study, the regulatory mechanism of miR-22-3p/AKT3 in the development of Wilms' tumor (WT) was investigated. PATIENTS AND METHODS: Twenty-seven pairs of surgical tumor specimens and adjacent normal tissues were obtained from Jining No. 1 People's Hospital. The expression level of miR-22-3p in WT tissues and cell lines was measured by quantitative RT-PCR. MTT and transwell assays were performed to analyze cell proliferation and invasion in WT. The relationship between miR-22-3p and AKT3 was verified by a Dual-Luciferase assay. The protein expression of AKT3 was evaluated by Western blotting analysis. RESULTS: MiR-22-3p was downregulated and AKT3 was upregulated in WT. Functionally, overexpression of miR-22-3p inhibited cell proliferation and invasion in WT. Moreover, miR-22-3p directly targets AKT3. The knockdown of AKT3 suppressed cell proliferation and invasion in WT. In addition, upregulation of AKT3 restored the tumor suppressive effect of miR-22-3p in WT. CONCLUSIONS: MiR-22-3p inhibits the proliferation and invasion of WT cells by downregulating AKT3, indicating that miR-22-3p may be developed as a new biomarker for the diagnosis of WT.

Potential effects of IL-17A rs2275913 and IL-17F rs763780 polymorphisms on susceptibility to gastric cancer in Chinese population: a meta-analysis.

OBJECTIVE: This meta-analysis aims to clarify the effect of IL-17 polymorphisms on the susceptibility to GCa in the Chinese population. MATERIALS AND METHODS: Relevant pieces of literature were searched in PubMed, Web of School, VIP, and CNKI using the key words as "IL-17, gastric/stomach cancer" or "IL-17 polymorphisms, gastric/stomach cancer susceptibility". The odds ratio (OR) and 95% confidence interval (CI) in the selected studies were calculated using RevMan5.3 and STATA12.0. RESULTS: A total of 12 investigations reporting mutations in IL-17A rs2275913 and IL-17F rs763780 were enrolled. There were 11 studies reporting rs2275913 G>A, involving 3299 cases of GCa patients and 3339 cases of healthy controls. The random-effects model was performed since the heterogeneity test results of the recessive genetic model (GG&GA vs. AA) and the allelic model (G vs. A) of IL-17A rs2275913 G>A were I2>66%/p=0.001. Meanwhile, the dominant genetic model (GG vs. GA&AA) and the super-dominant genetic model (GA vs. GG&AA) of IL-17A rs2275913 G>A were I2< 50%/p>0.05, and the fixed-effects model was used. The meta-analysis showed that IL-17A rs2275913 G>A was positively correlated with GCa susceptibility under four genetic models (p<0.05). Five studies reporting IL-17F rs763780 T>C were enrolled, including 2535 cases of GCa patients and 2402 cases of healthy controls. The heterogeneity test showed that, except for the super-dominant genetic model, the p-value was <0.00001 in the dominant, recessive, and allelic models, and their I2 values were 87%, 88%, and 93%, respectively. Hence, a random-effects model was selected. IL-17F rs763780 T>C was positively correlated with GCa susceptibility under the super-dominant genetic model (p=0.003), rather than the other three models (p>0.05). CONCLUSIONS: IL-17A rs2275913 G>A polymorphism contributes to susceptibility to GCa in the dominant, recessive, allelic, and super-dominant models. Meanwhile, IL-17F rs763780 T>C polymorphism is positively correlated with GCa susceptibility in the super-dominant model.

Simvastatin promotes osteogenic differentiation of mesenchymal stem cells in rat model of osteoporosis through BMP-2/Smads signaling pathway.

OBJECTIVE: By establishing osteoporosis (OP) model in rats, the specific regulatory effect of simvastatin on promoting the differentiation of mesenchymal stem cells (MSCs) into osteoblasts through the bone morphogenetic protein 2 (BMP-2)/Smads signaling pathway was investigated. MATERIALS AND METHODS: A total of 45 Sprague-Dawley rats were selected to establish the OP model by performing ovariectomy. The rats were divided into OP model group (OP group, n=15), 10-7 mmol/L simvastatin treatment group (SIM group, n=15), and normal control group (Control group, n=15). After the experimental period, the enzyme-linked immunosorbent assay (ELISA) was applied to observe the serum levels of tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), and IL-1. Reverse Transcription-Polymerase Chain Reaction (RT-PCR) was adopted to detect the contents of the differentiation-associated genes [runt-related transcription factor 2 (RUNX2) and Osterix (Osx)]. Later, the bone marrow MSCs (BMSCs) were selected and divided into Control group, 10-7 mol/L simvastatin group (SIM group), and osteoinduction medium group (OM group). Cell morphology in each group was observed. The Cell Counting Kit-8 (CCK-8) was performed to determine the proliferation activity of BMSCs. ELISA was performed to measure the level of alkaline phosphatase (ALP). RT-PCR was conducted to examine the levels of key differentiation-associated gene RUNX2 and those in BMP-2/Smads pathway. Moreover, the Western blotting was adopted to analyze the expressions of RUNX2 and genes in BMP-2/Smads pathway. RESULTS: The serum levels of TNF-α, IL-6, and IL-1 in OP group were remarkably higher than those in the Control group, and their levels in the SIM group were close to those in the Control group. The elevated messenger ribonucleic acid (mRNA) levels of the key differentiation-associated factors RUNX2, osteoprotegerin (OPG), osteopontin (OPN), and Osx were observed in the SIM group. In vitro cell culture revealed that the cells were in a favorable growth status in the SIM group and OM group, mostly manifesting in fusiform or spindle shape, and proliferated rapidly. In addition, the ALP level notably increased in the two groups compared with that in the Control group (p<0.05). Both SIM group and OM group had evidently higher mRNA expression levels of RUNX2, OPG, OPN, and Osx than those in the Control group (p<0.05), consistent with the expression trends of the genes in BMP-2/Smads pathway. The Western blotting indicated that the expression levels of RUNX2 and genes in BMP-2/Smads pathway in the SIM group were significantly higher than those in the Control group. CONCLUSIONS: Simvastatin can promote the differentiation of MSCs into osteoblasts in the OP rat model through the BMP-2/Smads signaling pathway.

Dexamethasone alleviates allergic asthma immature rat through Toll like receptor 4.

OBJECTIVE: The allergic asthma model induced by ovalbumin (OVA) was established in the immature rat. Dexamethasone (DXM) was adopted for intervention to analyze the treatment effect and to explore the relationship with toll-like receptor 4 (TLR4). MATERIALS AND METHODS: Immature SD rat was treated by OVA to construct allergic asthma model and intervened by DXM. The rats were randomly divided into model group, experimental group, and control group. The changes in lung tissue were observed by light microscope. The EOS infiltration and reactivity of airway wall were compared. The expressions of TLR2 and TLR4 protein and mRNA in the lung tissue were tested by Western blot and RT-PCR. RESULTS: The lung tissue in the model group was infiltrated by a lot of inflammatory cells, and mucous membrane edema was observed, compared with that in the control group. There were only a few inflammatory cells in the interstitial tissue and pulmonary alveoli in the experimental group compared with that in the model group. EOS count of airway wall and airway reactivity decreased in the experimental group. The levels of TLR2 and TLR4 were significantly elevated in the third week compared with the first week (p<0.05). CONCLUSIONS: The treatment of DXM can alleviate the pathological changes of the lung tissue in SD immature rat with allergic asthma, reduce EOS infiltration in the airway wall, decrease airway reactivity, and elevate expressions of TLR2 and TLR4.

Knockdown of PTTG1 inhibits the growth and invasion of lung adenocarcinoma cells through regulation of TGFB1/SMAD3 signaling.

Increased expression of pituitary tumor-transforming gene 1 (PTTG1) is expressed in many tumors and regulates tumor growth and progression. However, the precise function of PTTG1 in the tumorigenesis of lung adenocarcinoma (LAC) is not defined yet. Here, we examined the expression of PTTG1 in human LAC tissues by immunohistochemical assay using a tissue microarray procedure. A loss-of-function experiment was carried out to investigate the effects of lentiviral vector-mediated PTTG1 shRNA (shPTTG1) on cell growth and invasive potential in LAC cell lines (A549 and LETPα-2), assessed by MTT and Transwell assays. As a consequence, we found that the expression of PTTG1 protein was markedly upregulated in LAC tissues compared with the adjacent non-cancerous tissues (ANCT) (54.0% vs. 28.0%, P = 0.008), and was positively associated with the lymphatic invasion of the tumor (P = 0.01). Moreover, knockdown of PTTG1 expression inhibited tumor proliferation and invasion of LAC cells, companied by the decreased expression of CyclinD1 and MMP-2 and increased expression of p-TGFß1 and p-SMAD3. Collectively, our findings indicate that high expression of PTTG1 is correlated with the tumor metastasis of LAC patients, and knockdown of PTTG1 suppresses the growth and invasion of LAC cells through upregulation of the TGFß1/SMAD3 signaling, suggesting that PTTG1 may be a potential target for developing an effective immunotherapeutic strategy for LAC.

Melting of multilayer colloidal crystals confined between two walls.

Video microscopy is employed to study the melting behaviors of multilayer colloidal crystals composed of diameter-tunable microgel spheres confined between two walls. We systematically explore film thickness effects on the melting process and on the phase behaviors of single crystal and polycrystalline films. Thick films (>4 layers) are observed to melt heterogeneously, while thin films (≤4 layers) melt homogeneously, even for polycrystalline films. Grain-boundary melting dominates other types of melting processes in polycrystalline films thicker than 12 layers. The heterogeneous melting from dislocations is found to coexist with grain-boundary melting in films between 5- and 12-layers. In dislocation melting, liquid nucleates at dislocations and forms lakelike domains embedded in the larger crystalline matrix; the "lakes" are observed to diffuse, interact, merge with each other, and eventually merge with large strips of liquid melted from grain boundaries. Thin film melting is qualitatively different: thin films homogeneously melt by generating many small defects which need not nucleate at grain boundaries or dislocations. For three- and four-layer thin films, different layers are observed to have the same melting point, but surface layers melt faster than bulk layers. Within our resolution, two- to four-layer films appear to melt in one step, while monolayers melt in two steps with an intermediate hexatic phase.

Involvement of tumor suppressor in lung cancer 1 gene expression in cervical carcinogenesis.

Tumor suppressor in lung cancer 1 (TSLC1) is a tumor suppressor gene in non-small cell lung cancer, and loss of TSLC1 gene expression has been observed in a number of epithelial carcinomas and cancer-derived cell lines. We analyzed TSLC1 gene expression by real-time reverse transcription-polymerase chain reaction in 39 invasive cervical carcinomas, 34 cervical intraepithelial neoplasia (CIN) IIIs, 35 CIN IIs, 32 CIN I, 36 inflammation cervical tissues, and 30 normal cervix samples. Loss of TSLC1 gene expression was observed in 30 of 39 (77%) cervical carcinomas, 25 of 34 (73%) CIN IIIs, 9 of 35 (26%) CIN IIs, and 7 of 32 (22%) CIN Is but was not found in inflammation and normal cervix samples. Compared to normal cervical tissue, loss of TSLC1 gene was significantly high in CIN IIIs and cervical cancer (P = 0.00). Moreover, loss of TSLC1 gene expression is observed at a significantly higher frequency in CIN IIIs and cervical cancers than in CIN IIs (P < 0.05). The results show that loss of TSLC1 gene expression is an early event in cervical carcinogenesis and often accompanies invasive cervical cancers.