Circ_ANKIB1 stabilizes the regulation of miR-19b on SOCS3/STAT3 pathway to promote osteosarcoma cell growth and invasion.

Osteosarcoma is a highly malignant tumor. The molecular mechanism of its occurrence and development has not yet been clarified. Current studies have found that noncoding RNAs, such as circular RNAs (circRNAs) and microRNAs (miRNAs), play important regulatory roles in the progression of diseases. Our previous studies have shown that miR-19b is an oncogene in osteosarcoma. Further studies have shown that circ_ANKIB1 has binding sites for miR-19b, and both molecules were generally upregulated in osteosarcoma cells. RIP assay, RNA pull down, and dual-luciferase reporter gene assay showed that circ_ANKIB1 could directly bind to miR-19b and act as an miR-19b sponge in osteosarcoma cells. Circ_ANKIB1 promoted miR-19b expression, inhibited the expression of the downstream target gene SOCS3, and then activated the STAT3 pathway. When cotransfected with circ_ANKIB1 siRNA, and miR-19b mimics, the expression of SOCS3 and the phosphorylation level of STAT3 did not change significantly. Continuous detection of cell growth and invasion showed that the downregulation of circ_ANKIB1 or miR-19b significantly inhibited cell proliferation and invasion, but increased the apoptotic level. When cotransfected with circ_ANKIB1 siRNA and miR-19b mimics or SOCS3 siRNA, the cell proliferation, apoptosis, and invasion levels did not change significantly, suggesting that circ_ANKIB1 could affect the STAT3 pathway and osteosarcoma cell growth and invasion by enhancing the regulation of miR-19b on the downstream target gene SOCS3. These findings suggest that circRNAs stabilize miRNA functions, and further studies on this new function of circRNAs will provide a meaningful reference for the diagnosis and treatment of tumors and other diseases.

[Biological effects of root exudates from resistant and susceptible banana varieties on Fusaiurm oxysporum f. sp. cubense and Bacillus subtilis]. / 香蕉抗(感)ç— å“ç§æ ¹ç³»åˆ†æ³Œç‰©å¯¹æž¯èŽç— èŒå’Œæž¯è‰èŠ½å­¢æ†èŒçš„ç”Ÿç‰©æ•ˆåº”.

Root exudates of banana resistant variety ('Nantianhuang') and susceptible variety ('Guijiao No. 6') to Fusarium wilt were collected in vitro by bathing root system to examine the biological effects of root exudates from banana varieties on Fusaiurm oxysporum f. sp. cubense and Bacillus subtilis. We explored the effects of root exudates of different banana varieties on the abundance of soil microorganisms and the growth of F. oxysporum f. sp. cubense and B. subtilis. The results showed that root exudates from resistant variety could significantly reduce the abundance of soil fungi and inhibit the spore germination of F. oxysporum f. sp. cubense. Root exudates from susceptible variety could significantly stimulate mycelia growth and spores germination, whereas root exudates from the tested banana varieties could significantly increase the growth and biofilm formation of B. subtilis. By dealing with the root exudates of resistant and susceptible varieties, the growth rate of mycelia were 11.28 and 12.28 mm·d-1, and the germination rate of spores were 34.6% and 79.5%, respectively. After culturing for 12 h, the growth rates of B. subtilis (OD600) were 1.27 and 1.14, and the biofilm formation (OD570) were 1.11 and 1.30 after static culturing 72 h, respectively. There were significant differences between the values of resistant and susceptible varieties. The colonization amount of B. subtilis in the rhizosphere of susceptible variety was significantly higher than that of resistant variety. The contents of free amino acids and organic acid in root exudates of the resistant variety were higher than that of susceptible variety. The content ratio of acetic acid and proline in the root exudates of resistant variety were 3.7 times and 2.4 times of that of susceptible variety. In conclusion, root exudates of banana resistant variety could inhibit the growth of F. oxysporum f. sp. cubense. Root exudates from susceptible variety could promote the growth of F. oxysporum f. sp. cubense,while that from the tested banana varieties could all significantly enhance growth, biofilm formation and colonization ability of B. subtilis.

Development of a dendrimer PAMAM­based gold biochip for rapid and sensitive detection of endogenous IFN­Î³ and anti­IFN­Î³ IgG in patients with hemophagocytic lymphohistiocytosis.

Hemophagocytic lymphohistiocytosis (HLH) is a rare but severe disease characterized by immune hyperactivation and cytokine storm. Given the high mortality rate of HLH, there is a need for more effective diagnostic tools and treatments. The present study developed a dendrimer­based protein biochip for rapid, sensitive and simultaneous detection of serum interferon (IFN)­Î³ and endogenous anti­IFN­Î³ antibody (Ab) in patients with HLH. A gold biochip was modified with 1, 4­phenylene diisothiocyanate (PDITC), polyamidoamine (PAMAM) or PDITC­activated PAMAM. The optimal immobilization concentration for Ab capture and the reaction concentration for detecting Ab on the PDITC­activated PAMAM­modified biochip were 6.25 and 3.12 µg/ml, respectively; the limit of detection of IFN­Î³ protein was 50 pg/ml. The efficiency of the protein­probed biochip in detecting IFN­Î³ and anti­IFN­Î³ Ab in serum samples from 77 patients with HLH was evaluated; the positive rates for IFN­Î³ and anti­IFN­Î³ IgG Ab were 63.6% (49/77) and 61.0% (47/77), respectively. The present results demonstrated that the PDITC­activated PAMAM­modified biochip might be a sensitive tool for the specific detection of IFN­Î³ and anti­IFN­Î³ Ab in serum, and might have clinical applicability for the diagnosis of HLH.

[Effect of minimally invasive plate osteosynthesis through two approaches on bone metabolic activity and radial nerve injury in patients with humeral midshaft fracture].

OBJECTIVE: To compare application of minimally invasive plate osteosynthesis (MIPO) through two approaches for patients with humeral midshaft fracture, and analyzes the effect on bone metabolic activity and radial nerve injury. METHODS: From April 2014 to May 2017, 76 patients with humeral midshaft fracture treated by MIPO were selected and randomly divided into group A (anterior approache group) and group B (lateral approach group) according to random number stable. In group A, there were 38 patients including 22 males and 16 females, aged from 18 to 74 years old with an average age of(48.21±5.79) years old; 24 patients were caused by traffic accidents, 6 patients were caused by heavy object crushing, 8 patients were caused by falling down; 20 patient were on the right side, 18 patients were on the left side; 15 patients were type A, 17 patients were type B, 6 patients were type C according to AO classification; the patients were treated by MIPO through anterior approach. In group B, there were 38 patients including 23 males and 15 females, aged from 20 to 73 years old with an average age of(48.40±5.81) years old; 26 patients were caused by traffic accidents, 5 patients were caused by heavy object crushing, 7 patients were caused by falling down; 17 patients were on the right side, 21 patients were on the left side; 15 patients were type A, 18 cases were type B, 5 cases were type C according to AO classification; the patients were treated by MIPO through lateral approach. Intraoperative blood loss, operative time, hospital stays, fracture healing time between two groups were compared. Bone gla protein (BGP), collagen C-terminal peptide (CTX) and osteoprotegerin (OPG) were tested before and after operation. The incidence of radial nerve injury after operation was observed. RESULTS: All patients were followed-up from 12 to 18 months with an average of (15.4±2.1) months. There were no statistical differences in operative time, intraoperative blood loss between two groups. Hospital stays, fracture healing time in group A were(6.52±1.81) d, (13.27±3.01) weeks respectively, while in group B were(9.61±1.99) d, (14.83±3.08) weeks; and had differences between two groups. There were no differences in BGP, OPG and CTX, BGP between two groups and OPG in group A at 1 month after operation were(7.10±0.58) ng/ml, (173.67±9.12) pg/ml and higher than that of group B(6.63±0.62) ng/ml, (152.80±9.23) pg/ml; while CTX in group A (224.52±12.67) µg/ml was lower than that of group B(259.13±13.54) µg/ml(P<0.05). No patient occurred radial nerve injury in group A, 4 patients occurred radial nerve injury in group B, and had statistical differences between two groups(χ²=4.220, P<0.05). CONCLUSIONS: Compared with lateral approach, anterior approach has much more effective in minimally invasive MIPO for humeral shaft fractures, which could improve bone metabolism and reduce risk of radial nerve injury.

A biochip-based combined immunoassay for detection of serological status of Borrelia burgdorferi in Lyme borreliosis.

BACKGROUND: Dithiobis (succinimidyl undecanoate) modified gold surface biochip were used as a combined immunoassay platform for concurrently detecting immune responses to Borrelia burgdorferi (B. burgdorferi) sensu lato antigens, flagellin, outer surface protein C, variable major protein-like sequence proteins, and 3 VlsE protein IR6 peptides. The peptides represented intrinsic Borrelia genospecies: B. burgdorferi sensu stricto, B. garinii, and B. afzelii, respectively. METHODS: Fourier transform infrared spectroscopy was utilized to validate the surface chemical characteristics on the modified gold surface. RESULTS: The limits in detection of IgG antibody on the biochips were as little as 0.39µg/ml for anti-VlsE and 0.78µg/ml for anti-flagellin and anti-OspC, respectively. Samples from 56 neuroborreliosis (NB) patients and 114 healthy individuals were analyzed by the combined biochip. We found that the seroprevalences of IgM or IgG antibody against the 6 antigens were contributed to increased overall sensitivity by the multiplex immunobiochip assay. Serum combined positive rates of the 6 antigens in the patients were 92.86% for IgM antibody and 91.07% for IgG antibody. Part of the patients bore antibody responses against the 3 VlsE IR6 variant peptides, indicating that Lyme borreliosis would attribute to consequence of multiple infections by one or more Borrelia burgdorferi strains. Concurrent assessment for both IgM and IgG antibodies against the protein antigens and B. burgdorferi IR6 peptides in the sera of NB patients was beneficial from the biochip format, enabling detection of expanded serologic infection status and therapy strategy-making more efficiently. CONCLUSIONS: The combined biochip-based immunoassay, as a potential substitution of ELISA, provided a promising approach to extend the detection spectrum of infectious antibodies against a panel of Borrelia antigens.

Succinyl-ß-cyclodextrin modified gold biochip improved seroimmunological detection sensitivity for Lyme disease.

Cyclodextrin (CD) is a kind of cyclic oligosaccharides, which forms host-guest interactions with hydrophobic molecules and is widely applied in capillary electrophoresis and pharmaceutical engineering. In this study, we established a succinyl-ß-CD modified gold biochip for improvement of seroimmunological detection sensitivity of Lyme disease. We found that the CD modified biochip platform presented a stronger affinity property for VlsE protein in conjugation with >0.000475 µg/mL of antigen immobilization concentration, which was sensitive enough for fluorescence based assay. Detection limit for anti-VlsE IgG antibody was 0.39 µg/mL. Specificity of VlsE assay on the succinyl-ß-CD modified biochip was successfully confirmed by an immunological block assay. Furthermore, the correlation coefficient (R2) between the fluorescence values by the biochip and the OD values by ELISA assay was 0.904, indicating this biochip-based immunological assay might be a potential substitute with the ELISA-based approach. This biochip platform would be not suitable for loading of flagellin and OspC.

Development of a novel protein biochip enabling validation of immunological assays and detection of serum IgG and IgM antibodies against Treponema pallidum pathogens in the patients with syphilis.

In this study, we developed a novel protein biochip methodology that was characterized by dithiobis (succinimidyl undecanoate) (DSU) and specialized for detection of serum IgG and IgM antibodies against Treponema pallidum pathogens in the patients with syphilis, respectively. The biochips were validated by a dimension of atomic force microscope (AFM). The visualized detection limit of IgG antibody on the biochip was 0.39µg/ml. Finally, 286 serum samples from the patients with syphilis were simultaneously tested on the rTpN15-17-47 coated biochips. The results were evaluated in comparison with the assays of T. pallidum particle agglutination (TPPA) and the toluidine red unheated serum test (TRUST). The result demonstrated that the relative positive rate in the 286 patients by biochip was 99.0%, similar to that by TPPA (97.9%, P>0.05) and higher than that by TRUST, (76.2%, P<0.01). The detection specificities were 100% for the biochip and the TPPA and 97.0% for the TRUST. Thus, the protein biochip would provide a useful platform not only for enabling concurrent detection of the infectious antibodies directed against T. pallidum on a larger scale, but also for monitoring therapy modality of the disease.

Genetic association of osteopontin (OPN) and its receptor CD44 genes with susceptibility to Chinese gastric cancer patients.

PURPOSE: (1) To investigate associations between single nucleotide polymorphisms (SNPs) in osteopontin (OPN) and its receptor-cluster of differentiation 44 (CD44) genes and gastric cancer susceptibility. (2) To explore the correlation of OPN and CD44 expression of gastric cancer. METHODS: We detected 26 SNPs of the genes in gastric cancer patients from the Chinese Han population by Sequenom technique and performed expression of OPN in combination with CD44 in 243 tissues samples of the cases by tissue microarray and immunohistochemistry (IHC). RESULTS: We found that the minor alleles of OPN rs4754C>T and OPN rs9138C>A remained strongly associated with decreased gastric cancer risk (P = 1.53 × 10(-4), odds ratio (OR) 0.642, 95 % confidence interval (CI) 0.511-0.808 and P = 1.59 × 10(-4), OR 0.642, 95 %CI 0.510-0.809). OPN variant rs1126772A>G and CD44 variant rs353639A>C significantly contributed to elevated risk of gastric cancer (P = 0.042, OR 1.279, 95 % CI 1.008-1.622 and P = 0.047, OR 1.334, 95 % CI 1.003-1.772). Haplotypes of OPN and CD44 variants significantly influenced risk of gastric cancer. Clinical data indicated that rs4754 and rs9138 of OPN were significantly associated with smoking (P = 0.029, OR 0.343, 95 % CI 0.127-0.926 and P = 0.029, OR 0.343, 95 %CI 0.127-0.926) and OPN rs1126772 revealed associations with tumor-node-metastasis (TNM) stage (P = 0.025, OR 1.765, 95 % CI 1.073-2.905) and tumor differentiation (P = 0.031, OR 1.722, 95 % CI 1.049-2.825). OPN expression was observed in 133 of the 243 cases (54.7 %) by IHC and was correlated with serosa invasion (P = 0.013), TNM stage (P = 0.003) and lymph node metastasis (P = 0.002). CD44 expression was found in 92 of the 243 cases (37.9 %) and was associated with tumor size (P = 0.005) and lymph node metastasis (P = 0.023), respectively. The OPN expression displayed a positive association with CD44 (P = 0.01, r s = 0.164). CONCLUSIONS: We found that the polymorphisms rs4754, rs9138 and rs1126772 of OPN gene and rs353639 of CD44 gene were significantly associated with gastric cancer. Our IHC data indicated that interaction of OPN and CD44 protein would promote progression and metastasis of gastric cancer.