CircRNF20 aggravates the progression of non-small-cell lung carcinoma by activating MAPK9.

OBJECTIVE: To explore the clinical significance of circRNF20 in non-small-cell lung carcinoma (NSCLC), and its regulatory effects on NSCLC cell functions by activating MAPK9. PATIENTS AND METHODS: Relative levels of circRNF20 and MAPK9 in NSCLC tissues were detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). The relationship between circRNF20, MAPK9 and pathological factors in NSCLC patients was analyzed. Prognostic potentials of circRNF20 and MAPK9 in NSCLC were assessed by Kaplan-Meier method. The interaction between circRNF20 and MAPK9 was tested by Dual-Luciferase reporter assay. Regulatory effects of circRNF20 and MAPK9 on proliferative abilities in H358 and SPC-A1 cells were examined by Cell Counting Kit-8 (CCK-8) and colony formation assay. RESULTS: CircRNF20 and MAPK9 were upregulated in NSCLC tissues than normal ones. They were correlated to T stage and poor prognosis in NSCLC patients, while their levels were unrelated to gender, age, and incidences of lymphatic and distant metastasis. Knockdown of circRNF20 attenuated proliferative abilities in H358 and SPC-A1 cells. On the contrary, the overexpression of MAPK9 yielded the opposite results. MAPK9 was the target gene binding circRNF20, which was able to reverse the regulatory effect of circRNF20 on NSCLC proliferation. CONCLUSIONS: CircRNF20 and MAPK9 are upregulated in NSCLC cases, which are closely linked to T stage in NSCLC patients. They are independent prognostic factors for NSCLC. By activating MAPK9, circRNF20 stimulates NSCLC proliferation.

Effects of lncRNA SNHG20 on proliferation and apoptosis of non-small cell lung cancer cells through Wnt/ß-catenin signaling pathway.

OBJECTIVE: To investigate the influence of long non-coding ribonucleic acid (lncRNA) small nucleolar host gene 20 (SNHG20) on the proliferation and apoptosis of non-small cell lung cancer (NSCLC) cells through the Wnt/ß-catenin signaling pathway. PATIENTS AND METHODS: The human NSCLC cells were cultured and lncRNA SNHG20 was inhibited using si-SNHG20 and overexpressed using SNHG20-OE. Then, flow cytometry was used to detect the apoptotic rate. The targets of lncRNA SNHG20 were detected via dual-luciferase reporter gene assay, and the changes in the protein level were detected via Western blotting. RESULTS: LncRNA SNHG20 was highly expressed in the cancer tissues and serum of patients with NSCLC. LncRNA SNHG20 could promote the proliferation and inhibit the apoptosis of NSCLC cells. LncRNA SNHG20 could bind to micro RNA (miR)-197 in a targeted manner. Besides, nuclear translocation of ß-catenin was significantly enhanced after transfection of miR-197. After the down-regulation of miR-197 by small interfering RNA (siRNA), the key molecules TCF and LEF1 of the Wnt/ß-catenin pathway were significantly down-regulated. CONCLUSIONS: LncRNA SNHG20 promotes the proliferation and inhibits the apoptosis of NSCLC cells by targeting miR-197 through the Wnt/ß-catenin signaling pathway.

[A retrospective analysis of 200-cases of otosclerosis].

Objective:To identify the audiological improvement postoperatively and improve the understand of otosclerosis for a better performance of personalized surgical treatment.Method:We retrospectively reviewed a series of 200 cases after surgery for ototsclerosis.The clinical characteristics,pre-and postoperative audiometric results,signs of Carhart notch, Gelle test and the characteristics of high-resolution computed tomography of temporal bone were analyzed retrospectively.Result:73% of patients had tinnitus, while 4% had sense of ear fullness,80.79% had Carhart notch,92.09% had negative result in Gelle test and 45.66% had positive signs on computed tomography. 68% of the patients revealed a type A tympanogram with only 22% type As.Fifty-six cases with laser stapedotomy achieved a the air bone gap at 250 Hz,500 Hz,1 kHz,2 kHz and 4 kHz of 25.54,16.25,13.75,6.34,15.96 dB,respectively. The bone conduction thresholds at 250 Hz,500 Hz,1 kHz,2 kHz improved 2.05,1.51,3.75 and 3.93 dB,respectively. At 4 kHz, bone conduction threshold increased by 1.34 dB.The improvement of bone conduction threshold at 250 Hz,1 kHz,2 kHz was significantly but for the revisions at 500 Hz and 4 kHz.Conclusion:The diagnosis of otosclerosis should be based on the combination of medical history, pure tone audiometry, tympanometry, Carhart notch, Gelle test and high resolution computed tomography of temporal bone.Surgical technique of stapedotomy with Piston artificial auditory ossicle implantation could improve not only the air conduction threshold, but also the bone conduction threshold at 250 Hz,1 kHz,2 kHz.

Infusible platelet membrane microvesicles: a potential transfusion substitute for platelets.

BACKGROUND: Several substitutes for intact, viable platelets have been used for transfusion, both to people and in animal models, with varied success. Infusible platelet membrane (IPM) is prepared from human platelets. IPM retains the glycoprotein (GP)lb receptor and has platelet factor 3 activity (procoagulant activity). However, factor V, serotonin, a cytoplasmic marker enzyme (purine nucleotide phosphorylase), GPIIb/IIIa complex, and HLA class I and II antigens are all absent in IPM. STUDY DESIGN AND METHODS: IPM is prepared from outdated platelets. The platelets were disrupted by freezing and thawing; they were washed and heated to inactivate possible viral contaminants, and then the sonicated membrane microvesicle fraction was separated and lyophilized. The hemostatic activity of IPM was measured by its ability to reduce the prolonged bleeding time in thrombocytopenic rabbits. RESULTS: Administration of IPM at a dose of 2 mg per kg results in a substantial reduction in the bleeding time. In a series of 23 experiments, a median preinjection bleeding time of 15 minutes was reduced to 6 minutes within 4 hours after IPM administration. Administration of IPM did show a mild enhancement in the thrombogenicity index, as measured in the Wessler rabbit model. This enhancement is, however, not significant, as a thrombogenicity index value of up to 0.6 is clinically acceptable. CONCLUSION: IPM may have clinical potential as a substitute for platelets in the treatment of bleeding due to thrombocytopenia.

The use of diaminodithiol for labeling small molecules with technetium-99m.

To investigate the labeling of small molecules with 99mTc by the bifunctional chelate approach, we have synthesized both a fatty acid and an estrone derivative containing a chelator of the N2S2 type. In the case of the fatty acid, this was a diaminodithiol (DADT) while for the estrone, a diaminodisulfide (DADS) was attached. The estrone derivative (5-(2-methylene estrone 3-methyl ether)-3,3,10,10-tetramethyl-1,2-dithia-5,8-diazacyclodecane hydrochloride, DADS-E) was prepared by alkylation of DADS while the fatty acid derivative (N-(11-undecanoic acid)-N,N'-bis(2-methyl-2-mercaptopropyl) ethylenediamine hydrochloride, DADT-FA) was synthesized by alkylation of DADS followed by reduction. DADS-E was labeled in ethanol at elevated temperatures while DADT-FA was labeled at room temperature, both by stannous reduction. Paper chromatography showed both to be labeled and reverse-phase HPLC showed multiple peaks for both. Serum stability studies were performed by incubation at 37 degrees C with aliquots removed at 1 min and 1 day for analysis by size-exclusion HPLC. Initially, little pertechnetate or binding to serum proteins was observed whereas after 1 day the majority of activity in both cases was protein bound with 20 and 38% pertechnetate appearing for DADT-FA and DADS-E respectively. In conclusion, small biologically active molecules may be labeled with 99mTc through an attached diaminodithiol or diaminodisulfide group.

A new synthetic method for 99mTc labeled N-pyridoxyl-5-methyltryptophan as a hepatoma imaging agent.

N-Pyridoxyl-5-methyltryptophan (5-PMT) was synthesized by a simplified method using sodium borohydride for the reduction of a Schiff base of pyridoxal and 5-methyltryptophan. Lyophilized kits containing 5-PMT, stannous chloride and L-(+)-ascorbic acid were prepared and labeled to afford 99mTc-5-PMT with 96% or higher radiochemical purity analysed by two thin-layer chromatographic solvent systems. 99mTc-5-PMT showed a rapid blood clearance, a faster hepatobiliary transit and a lower renal retention in comparison with 99mTc-5-EHIDA in rats. Eleven (61%) of 18 patients with histologically confirmed hepatocellular carcinoma showed positive images at 2 to 5 h after i.v. injection. The smallest tumor that could be identified was 2 cm in diameter with the best tumor/liver ratio of 4. In conclusion, 99mTc-5-PMT synthesized by sodium borohydride reduction shows great promise as a useful hepatoma imaging agent.

Serum stability and non-specific binding of technetium-99m labeled diaminodithiol for protein labeling.

Previously we investigated the use of DTPA-coupled proteins to simplify labeling with 99mTc but especially to improve the stability of the label. These investigations have now been extended to include several N2S2 ligands such as N,N'-bis(2-methyl-2-mercaptopropyl)ethylenediamine (DADT) and a novel ligand of similar structure with a propylene bridge between two amines, 2-hydroxy-N,N'-bis(2-methyl-2-mercaptopropyl)propylenediamine++ + (DADT-3C-2OH). The condition of labeling of free ligand (pH, buffer and tin concentration) was optimized to provide 100% chelation with 99mTc at reasonable ligand concentrations (100 micrograms/mL or less). Labeling was determined by paper chromatography, reverse-phase and size-exclusion HPLC. After incubation in fresh serum, 37 degrees C for 24 h, repeat analysis showed less than 5% dissociation of the chelate. By contrast, the DTPA chelate shows instability towards oxidation during this period. DADT derivatized on an ethylene carbon showed almost identical serum stability as DADT itself whereas when derivatized on a nitrogen greater instabilities were apparent. Using identical labeling conditions, free DADT was chelated in the presence of IgG at different ligand: protein molar ratios. Non-specific binding of 99mTc to IgG at a 10:1 DADT-HM:IgG molar ratio was as little as 5% and was essentially zero at a 2:1 DADT:IgG molar ratio when labeling was by transcomplexation from 99mTc-EDTA. The DADT-3C-2OH ligand showed superior performance both in regard to serum stability and the absence of non-specific binding. In conclusion, the N2S2 ligands form more stable chelates with 99mTc than does DTPA with reduced non-specific binding and may therefore represent an attractive alternative for labeling proteins with 99mTc by the bifunctional chelate approach.