Effects of lncRNA ANRIL on proliferation and apoptosis of oral squamous cell carcinoma cells by regulating TGF-ß/Smad pathway.

OBJECTIVE: To investigate the role of the long non-coding ribonucleic acid (lncRNA) antisense non-coding RNA in the INK4 locus (ANRIL) in the proliferation and apoptosis of the oral squamous cell carcinoma (OSCC) cells by regulating the transforming growth factor-beta (TGF-ß)/Smad pathway. PATIENTS AND METHODS: Human OSCC cells were cultured, and then transfected with small interfering (si)-ANRIL to inhibit the lncRNA ANRIL and ANRIL-OE to overexpress the lncRNA ANRIL. Next, the flow cytometry was carried out to detect the apoptosis rate, the proliferation was determined via methyl thiazolyl tetrazolium (MTT) assay, and the changes in the protein level were detected through Western blotting (WB). RESULTS: The lncRNA ANRIL was highly expressed in the tissues and serum of patients. The proliferation ability of the cells transfected with si-ANRIL was significantly reduced, while that of the cells transfected with ANRIL-OE was overtly increased. The apoptosis rate was (9.21±5.22)%, (22.3±1.34)%, and (13.21±6.22)% in lncRNA ANRIL-OE group, si-ANRIL group and control group, respectively. The protein expression level of the apoptotic protein active caspase-3 was lowered after the treatment with ANRIL-OE, and the key molecules of the TGF-ß/Smad pathway were notably down-regulated after inhibiting ANRIL with si-ANRIL. CONCLUSIONS: The lncRNA ANRIL regulates the TGF-ß/Smad signaling pathway to promote the proliferation and suppress the apoptosis of OSCC cells.

A novel method to detect circulating antigens of Schistosoma japonicum using a gold nanorod optical sensor.

Diagnostic method of Schistosoma japonicum (S.japonicum) is the key of schistosomiasis prevention and control. In China, schistosomiasis reached the stage of transmission control, and almost of the epidemic areas tend to have low infection rate and intensity, but it is difficult for the existing detection methods to achieve accurate monitoring. In this study, a novel method to detect the circulating antigens of S.japonicum using gold nanorods optical sensor was developed. Gold nanorods were prepared by seed-mediated growth followed by deposition onto Indium-Tin-Oxide (ITO) glass to fabricate a solid phase biosensor. In order to assembly between the ITO glass and gold nanorod, hydroxylation and sulfhydrylation were carried out to modify the ITO glass. Surface of gold nanorods was conjugated with an SIEA26-28kDaSjscFv antibody against S.japonicum circulating antigens, and the sensor optical changed upon antigen-antibody recognition. The sensor was used to detect S.japonicum infection in rabbits by testing the serum once a week for 8 weeks. Results revealed different displacement of localized surface plasmon resonance (LSPR) of the gold nanorod optical sensor each week while the control group showed no such change in LSPR. Simultaneously, Indirect Hemagglutination Assay(IHA) and Fast Enzyme-Linked Immuno Sorbent Assay (F-ELISA) method were used to test these samples. Ten human serum samples from S.japonicum infected patients were analyzed using the gold nanorods optical sensor, which revealed that health human serum did not show any spectrum displacement. We developed a specificity gold nanorod optical sensor by combining the SIEA26-28kDaSjscFv, which was used to detect circulating antigens of S.japonicum. This method is expected to overcome the issues pertaining to the testing of circulating antigens of S.japonicum.

HCV-induced miR146a controls SOCS1/STAT3 and cytokine expression in monocytes to promote regulatory T-cell development.

Host innate and adaptive immune responses must be tightly regulated by an intricate balance between positive and negative signals to ensure their appropriate onset and termination while fighting pathogens and avoiding autoimmunity; persistent pathogens may usurp these regulatory machineries to dampen host immune responses for their persistence in vivo. Here, we demonstrate that miR146a is up-regulated in monocytes from hepatitis C virus (HCV)-infected individuals compared to control subjects. Interestingly, miR146a expression in monocytes without HCV infection increased, whereas its level in monocytes with HCV infection decreased, following Toll-like receptor (TLR) stimulation. This miR146a induction by HCV infection and differential response to TLR stimulation were recapitulated in vitro in monocytes co-cultured with hepatocytes with or without HCV infection. Importantly, inhibition of miR146a in monocytes from HCV-infected patients led to a decrease in IL-23, IL-10 and TGF-ß expressions through the induction of suppressor of cytokine signalling 1 (SOCS1) and the inhibition of signal transducer and activator transcription 3 (STAT3), and this subsequently resulted in a decrease in regulatory T cells (Tregs) accumulated during HCV infection. These results suggest that miR146a may regulate SOCS1/STAT3 and cytokine signalling in monocytes, directing T-cell differentiation and balancing immune clearance and immune injury during chronic viral infection.

Salt stress induces programmed cell death in prokaryotic organism Anabaena.

AIMS: Our main interest is to check if programmed cell death (PCD) can occur in prokaryotic algae and if the morphological and biochemical features of PCD are conserved. METHODS AND RESULTS: Using TUNEL labelling, fluorescence and light microscopy and DNA gel electrophoresis, we found that cell death with features similar to those in metazoan PCD could be induced in different Anabaena strains after exposure to univalent-cation salts at moderate concentration. These features included specific DNA fragmentation, cytoplasmic vacuolation, and the progressive disorganization, fragmentation and subsequent autolysis of the cell corpse. Further analyses of cell viability and proteinase activity revealed that increased protease activities, decreased DNA content, and loss of plasmalemma integrity were related to the PCD process. CONCLUSIONS: The results showed that like PCD in eukaryotes, PCD in Anabaena is an active process, and is an adaptation to adverse environments. The features of PCD shared between eukaryotes and Anabaena suggest that PCD mechanisms are conserved during evolution. SIGNIFICANCE AND IMPACT OF THE STUDY: The results will contribute greatly to our understanding of PCD origin and evolution, and are potentially useful in controlling the deluge of algae in some lakes.

[Detection and analysis of alien genes in transgenic rice by fluorescence in situ hybridization].

Using fluorescence in situ hybridization (FISH) with metaphase preparations, we localized a transferred barnase-psl DNA sequence onto chromosomes in 8 rice transgenic plants. All the tested rice transgenic lines showed hybridization signals on the middle and terminal regions of chromosome arms except for those close to centromeres. In two lines, two different integration sites were identified, and the other lines showed only one integration site. With the aid of Southern analysis and expression detection, we found that the barnase tended to show a higher level expression in the lines whose integration sites near the distal regions of chromosomes, while the expression level became lower in the lines whose integration sites near the centromeres. This result suggested a possible relationship between chromosomal location of transgenes and the expression level. However it showed no obvious relationship between copy numbers and expression levels. In most cases, the results of multi-color FISH showed that barnase-ps1 always integrated at the same position on the chrmosome as the reporter genes(pHctinG).

[Physical mapping of the genes px and cld coding peroxidase and cold-regulated protein in maize (Zea mays L.)].

Peroxidase plays a key role in plant disease resistance, cold stress and some developmental processes, and cold-regulated protein functions necessarily in reaction of plants on cold or heat stress. Recent studies showed that these processes in plant cells were involved in programmed cell death (PCD). Using a biotin-labelled in situ hybridization (ISH) technique, we physically mapped the genes px and cld coding peroxidase and cold-regulated protein respectively onto maize chromosomes. Both DAB and fluorescence detection systems gave the identical results, the probe uaz235 corresponding to gene px was localized onto the long arm of chromosome 2 (2L) and 7L, and csu19 corresponding to gene cld was hybridized onto 4L and 5L. The percentage distances (from the hybridization sites to centromeres) of uaz235 in 2L and 7L were 45.4 +/- 1.3 and 67.4 +/- 3.7 respectively, and those of csu19 in 4L and 5L were 68.6 +/- 2.6 and 58.2 +/- 1.6 respectively. The physical positions of px in 2L and cld in 4L coincide with those in their genetic map pattern. The results also show that both of these genes have duplicated sites in maize genome.

[Salt stress-induced apoptosis in plants--a possible resistance mechanism to salt stress].

Previous study has showed that apoptosis-like cell death can be induced under moderate salt stress in plants. In this paper, we studied the cell death induced by salt stress in maize, rice and tobacco roots utilizing DNA Laddering, paraffin sectioning-based TUNEL and chromosome spreading-based TUNEL simultaneously. The characteristic morphological and biochemical features showed apoptosis induced by salt stress may be a universal phenomenon in plants, but some differences may lie in various species. These results provided a valuable insight into studying the physiological mechanism of stress resistance in plants. In addition, we compared the in situ labelling technique based on chromosome spreading with that based on paraffin sectioning. We proposed according to the results that chromosome spreading-based in situ labeling technique should be suitable to detect individual cell death qualitatively and quantitatively with high efficiency, low background and detailed description of apoptotic changes at chromosome, nuclear and DNA levels.

Right ventricular size and ventricular septal motion after repair of atrial septal defect in children.

An enlarged right ventricle and abnormal ventricular septal motion are characteristic echocardiographic features of atrial septal defect and often persist after the defect has been completely closed, even when the operation clinically is judged to be successful. These features were examined retrospectively 15 to 21 months after operation in a group of children whose atrial septal defect had been closed between January 1976 and July 1979. Despite satisfactory postoperative results in all, about two thirds had an enlarged right ventricular dimension and about the same number had abnormal septal motion when examined echocardiographically an average of 18 months after operation. The best operative strategy seems to be to operate while the right ventricular end-diastolic dimension is still relatively small in echocardiographic terms.