MiR-21 regulates proliferation and apoptosis of oral cancer cells through TNF-α.

OBJECTIVE: MicroRNA (miRNA) can widely regulate gene expression. More importantly, various miRNA molecules have been found with regulatory functions for tumor cell proliferation or apoptosis. The study showed that miR-21 inhibited apoptosis of cultured cancer cells, whilst tumor necrosis factor α (TNF-α) plays important roles in the proliferation of tumor cells. This study manipulated miR-21 expression in cultured oral cancer cells and aimed to investigate its effects on TNF-α expression, and on proliferation or apoptosis of cancer cells. MATERIALS AND METHODS: Specific agonist and antagonist were synthesized based on miR-21 sequence. In vitro cultured oral cancer cell line, SCC-15 was transfected with agonist or antagonist, in parallel with normal cultured cells as negative control group. Quantitative Real-time PCR (qRT-PCR) was used to measure mRNA expression of miR-21 and TNF-α in transfected cells. Western blot was used for measuring TNF-α expression, and 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) or Hoechst-33342 staining was used to measure proliferation and apoptosis of SCC-15 cells. RESULTS: MiR-21 expression was potentiated or depressed with transfection of agonist or antagonist, respectively, illustrating the effectiveness of synthesized sequence in cultured SCC-15 cells. Moreover, TNF-α expression was positively correlated with miR-21, TNF-α up-regulation significantly potentiated the proliferation potency of SCC-15 cells, and TNF-α down-regulation remarkably weakened proliferation potency (p<0.05). The TNF-α expression did not affect apoptosis of SCC-15 cells (p>0.05 compared to the control group). CONCLUSIONS: MiR-21 could participate in the proliferation of cultured SCC-15 cells via targeting TNF-α expression, but without any significant effects on cell apoptosis.

Note: A simple feedback control method based on a real time acquisition voltage to avoid second breakdown of PFL.

To avoid serious damage in the pulse forming line (PFL) caused by repeated breakdown, a simple feedback control method based on a reverse peak voltage of the primary capacitor of the transformer is presented in this paper. The theoretical analysis of the breakdown circuit is carried out. The results indicate that once the breakdown occurs, the reverse peak voltage of the primary capacitor of the transformer increases obviously. A simple comparison function is added in the control system of the accelerator. If the collected reverse peak voltage of the primary capacitor is higher than the reference value, then the accelerator stops working immediately. The experimental result shows that this method can prevent the re-breakdown of the PFL effectively.

Expression and diagnostic value of proteins in Mycobacterium tuberculosis.

We constructed a prokaryotic expression vector expressing the Mycobacterium tuberculosis protein TB16.3, as well as 3 other proteins, including TB15.3, CFP-10, and Rv2626C, which were purified and analyzed for their effectiveness as detection antibodies. The TB16.3 genes of M. tuberculosis H37Rv genomic DNA were amplified by polymerase chain reaction, inserted into the expression vector pET-30a, and expressed in Escherichia coli. An enzyme-linked immunosorbent assay was used to detect the 4 M. tuberculosis antibodies. Engineered E. coli bacteria expressing TB16.3 and the 3 other proteins were constructed and found mainly to be soluble. For recombinant TB16.3 proteins, serum samples of 118 tuberculosis (TB) patients and 96 healthy controls were analyzed. Sensitivity, specificity, and adjusted concordance rate for the TB16.3 antibody were 72.9, 86.5, and 79.6%, respectively. The positive rate of Rv2626C antibody in TB patients (44.1%) was significantly lower than that in normal controls (75.0%, χ(2) = 20.8, P < 0.01). TB15.3 and TB16.3 were used for simultaneous detection and showed sensitivity, specificity, and repeatability rates of 69.4, 96.9, and 83.7%. The antibody positive rate and specificity for patients with lung disease was 9.6 and 90.4%, respectively. TB15.3 and TB16.3 were mixed and detected simultaneously. Combined with the results for TB15.3, the sensitivity, specificity, and concordance rates were 82.2, 95.9, and 88.9%, respectively. The concordance rate was the highest value observed. Target genes were cloned into a host strain and expressed successfully. The TB16.3 recombinant protein may be used as a new serological antigen for tuberculosis diagnosis.

Expression and serological diagnosis of Mycobacterium tuberculosis CFP-10 and Rv2626c proteins.

We constructed a Mycobacterium tuberculosis vector expressing CFP-10 and Rv2626c to examine the expression of these proteins in Escherichia coli as well as their immunoreactivity. The CFP-10 and Rv2626c genes were amplified from tuberculosis H37Rv genomic DNA using polymerase chain reaction. They were ligated into the expression vector PET30a and expressed in E. coli. Histidine tag nickel column chromatography was used to purify the recombinant protein. An enzyme-linked immunosorbent assay (ELISA) was used for detection. In our E. coli-engineered bacteria containing a CFP10 and Rv2626c plasmid, the target protein was found mainly to be in the soluble form. We formed mixed antigens of the recombinant CFP10 and Rv2626c proteins. ELISA results showed that in 214 blood samples, the positive rate was 77.1%. The target gene was successfully expressed in the host strain. Mixed antigens of the recombinant CFP-10 and Rv2626c proteins can be used as a combination antigen in the serological diagnosis of tuberculosis.

Identification of a new locus conferring antixenosis to the brown planthopper in rice cultivar Swarnalata (Oryza sativa L.).

The brown planthopper [Nilaparvata lugens (Stål); BPH] has caused severe damage to rice production. The identification of resistance genes and the development of BPH-resistant varieties are economical and effective ways to manage this pest. Using an F2 population from a cross between the Indica cultivars 93-11 and Swarnalata, we mapped the Qbph-8 locus to a 7.3-cM region on chromosome 8 in two tests, flanked by the markers RM339 and RM515. In this population, Qbph-8 explained 7.7 and 6.6% of the phenotypic variance of BPH preference in both tests. In the BPH host choice test, the average number of settled BPHs on the Qbph-8 plants was less than that on the 93-11 plants over the 24- to 120-h observation period. Furthermore, less BPH insects were observed on the BPH6+Qbph-8 plant compared with the BPH6 plant or Qbph-8 plant, indicating a stronger antixenotic effect shown in the gene pyramiding plants. Hence, this locus can be pyramided with other BPH resistance genes and applied to breed-resistant varieties, which possibly can improve the resistance level and durable resistance to the BPH.

An hsp70 fusion protein vaccine potentiates the immune response against Japanese encephalitis virus.

To evaluate the possibility of developing an effective subunit vaccine against Japanese encephalitis virus (JEV), mice were intraperitoneally immunized with either a neutralizing epitope (a 27-amino-acid region of the JEV E protein), or with a fusion protein between this region and a Mycobacterium tuberculosis hsp70. Both antigens were heterologously expressed in Escherichia coli as fusion proteins with thioredoxin. The fusion protein antigen elicited a higher titer of anti-thioredoxin-neutralizing epitope antibodies and a stronger proliferation of lymphocytes than did either the neutralizing epitope (irrespective of the presence of mineral oil as an adjuvant), or the conventional JEV SA14-14-2 vaccine. Assays of antibody isotype and IFN-gamma and IL-4 content in post-immunization serum showed that the fusion protein elicited a higher IgG2a titer and higher levels of IFN-gamma suggesting a potentiation of the Th1 immune response. The fusion protein antigen elicited a long-lived immune response, and the antibodies were able to neutralize JEV in vitro more strongly than did those elicited by the JEV SA14-14-2 vaccine. Immunization with the fusion protein generated both humoral and cellular immune responses to JEV, and the fusion protein appeared to be a more efficient protectant than the JEV SA14-14-2 vaccine.

[Heat transfer analysis of liquid cooling garment used for extravehicular activity].

Brief description was given about the construction and function of the LCG (liquid cooling garment) used for EVA (extravehicular activity). The heat convection was analyzed between ventilating gas and LCG, the heat and mass transfer process was analyzed too, then a heat and mass transfer mathematical model of LCG was developed. Thermal physiological experimental study with human body wearing LVCG (liquid cooling and ventilation garment) used for EVA was carried out to verify this mathematical model. This study provided a basis for the design of liquid-cooling and ventilation system for the space suit.

Preventive and therapeutic effects of nitrendipine on hypoxic right ventricular hypertrophy.

AIM: To assess whether nitrendipine (Nit) can be used to prevent and treat the hypoxic right ventricular hypertrophy (RVH). METHODS: Rats were exposed to a simulated altitude of 5000 m (barometric pressure = 54 kPa) for 30-60 d. Nit (10-20 mg.kg-1.d-1) was administered via gavage. The therapeutic efficacy was evaluated with right ventricular weight index (RVWI), right ventricular systolic pressure (RVSP), and myocardial ultrastructure. RESULTS: Chronic intermittent hypoxia for 30 d (8 h.d-1) resulted in an increase of RVSP and RVWI as well as in the changes of RV myocardial ultrastructure. As the hypoxic time was prolonged to 60 d, RVWI and RVSP were not further augmented. Nit (20 mg.kg-1.d-1, i.g.), when administered from the beginning of hypoxia, reduced RVSP (8.1 +/- 1.1 vs 6.0 +/- 1.0 kPa, P < 0.05) and RVWI (1.014 +/- 0.012 vs 0.915 +/- 0.049 mg/g body weight, P < 0.05). After development of hypoxic RVH, Nit (20 mg.kg-1) also decreased RVSP (7.9 +/- 1.0 vs 6.2 +/- 0.8 kPa, P < 0.05) and RVWI (1.02 +/- 0.13 vs 0.88 +/- 0.12 g/kg body weight, P < 0.05). Myocardial blood flow was increased and myocardial ultrastructure became nearly normal in rats treated with Nit. CONCLUSION: Nit prevented and lessened the hypoxic right ventricular hypertrophy.