Linoleic acid and α-linolenic acid inhibit conjugative transfer of an IncX4 plasmid carrying mcr-1.

AIMS: The aim of this study was to determine the effects of unsaturated fatty acids on clinical plasmids. METHODS AND RESULTS: Two unsaturated fatty acids, linoleic acid (LA) and α-linolenic acid (ALA) at final concentration 0, 0·03, 0·3 and 3 mmol l-1 , respectively, were used to assess the effects on conjugative transfer of a mcr-1-harbouring plasmid pCSZ4 (IncX4) in conjugation experiment. The inhibitory mechanisms were analysed by molecular docking and the gene expression of virB11 was quantitated by qRT-PCR. Target plasmid diversity was carried out by TrwD/VirB11 homology protein sequence prediction analysis. Our results showed that LA and ALA inhibit plasmid pCSZ4 transfer by binding to the amino acid residues (Phe124 and Thr125) of VirB11 with dose-dependent effects. The expression levels of virB11 gene were also significantly inhibited by LA and ALA treatment. Protein homology analysis revealed a wide distribution of TrwD/VirB11-like genes among over 37 classes of plasmids originated from both Gram-negative and Gram-positive bacteria. CONCLUSIONS: This study demonstrates representing a diversity of plasmids that may be potentially inhibited by unsaturated fatty acids. SIGNIFICANCE AND IMPACT OF THE STUDY: Our work reported here provides additional support for application of curbing the spread of multiple plasmids by unsaturated fatty acids.

The functions of caspase in whitefly Bemisia tabaci apoptosis in response to ultraviolet irradiation.

Whiteflies (Bemisia tabaci) are phloem feeders, and some invasive species are composed of cryptic species complexes that cause extensive crop damage, particularly via the direct transmission of plant viruses. Apoptosis is a type of programmed cell death essential for organismal development and tissue homeostasis. The caspases belong to a family of cysteine proteases that play a central role in the initiation of apoptosis in many organisms. Here, we employed a comprehensive genomics approach to identity caspases in B. tabaci Middle East Asia Minor 1 (MEAM1), an invasive whitefly that carries a cryptic species complex that is devastating to crops. Four caspase genes were identified, and their motif compositions were predicted. Structures were relatively conserved in both putative effector and initiator caspases. Expression patterns of caspase genes differed across insect developmental stages. Three caspase genes were induced immediately after ultraviolet (UV) treatment. Expression levels of Bt-caspase-1 and Bt-caspase-3b increased in the midgut and salivary glands during apoptosis induced by UV treatments, whereas silencing of both genes reduced UV-triggered apoptosis. Our study demonstrates that Bt-caspase-1 and Bt-caspase-3b, respectively, act as putative initiator and effector apoptotic caspases in the MEAM1 whitefly.

Horizontal gene transfer from a flowering plant to the insular pine Pinus canariensis (Chr. Sm. Ex DC in Buch).

Horizontal gene transfer (HGT) is viewed as very common in the plant mitochondrial (mt) genome, but, to date, only one case of HGT has been found in gymnosperms. Here we report a new case of HGT, in which a mt nad5-1 fragment was transferred from an angiosperm to Pinus canariensis. Quantitative assay and sequence analyses showed that the foreign nad5-1 is located in the mt genome of P. canariensis and is nonfunctional. An extensive survey in the genus Pinus revealed that the angiosperm-derived nad5-1 is restricted to P. canariensis and present across the species' range. Molecular dating based on chloroplast DNA suggested that the HGT event occurred in the late Miocene after P. canariensis split from its closest relatives, and that the foreign copy became fixed in P. canariensis owing to drift during its colonization of the Canary Islands. The mechanism of this HGT is unclear but it was probably achieved through either direct cell-cell contact or external vectors. Our discovery provides evidence for an important role of HGT in plant mt genome evolution.

Electroacupuncture inhibits excessive interferon-γ evoked up-regulation of P2X4 receptor in spinal microglia in a CCI rat model for neuropathic pain.

BACKGROUND: Although electroacupuncture (EA) is effective in the relief of neuropathic pain, the underlying mechanisms remain unclear. Previous studies have reported immunomodulatory effects of EA in rats. Since excessive release of interferon-γ (IFN-γ) after nerve injury transforms quiescent spinal microglia into an activated state with more neuropathic pain, associated with purinergic receptor P2X4 expression, it is possible that EA may mediate its analgesic effect by attenuating IFN-γ release and subsequent generation of P2X4R(+) microglia. METHODS: Male rats underwent chronic constriction injury (CCI) or IFN-γ intrathecal injection and von Frey tests were performed to evaluate the effect of EA on pain thresholds. Spinal IFN-γ and P2X4R expression levels were measured by immunohistochemistry, real-time PCR, enzyme immunoassay, and/or western blots. In vitro primary cultures of microglia were used to examine IFN-γ activation of P2X4R(+) cells. RESULTS: In CCI rats, EA treatment significantly increased paw withdrawal threshold relative to control. IFN-γ facilitated P2X4R(+) microglia activation both in vitro and in vivo. EA also down-regulated both P2X4R and IFN-γ expression in the spinal cord after CCI. However, EA did not exert the same analgesic effect after intrathecal IFN-γ injection. CONCLUSIONS: EA ameliorated tactile allodynia after peripheral nerve injury by down-regulating excessive expression of IFN-γ in the spinal cord and subsequently reducing expression of P2X4R.

Development of a DNA microarray-based multiplex assay of avian influenza virus subtypes H5, H7, H9, N1, and N2.

Outbreaks of highly pathogenic avian influenza have caused considerable economic losses in the poultry industry and have also resulted in human deaths since 2004. Rapid subtyping of highly pathogenic avian influenza viruses(HPAIVs) in clinical specimens is a prerequisite of prompt control of disease and prevention of its spreading. In this study, we describe development of a DNA microarray-based detection and subtyping of HPAIVs in field samples. DNA copies of matrix (M) protein genes for the H5, H7, and H9 subtypes of hemagglutinin (HA) and the N1 and N2 subtypes of neuraminidase (NA) were prepared by RT-PCR and specific primers and then spotted onto aldehyde slides to form DNA microarrays. The HPAIV samples to be tested were subjected to total RNA isolation, RT-PCR with universal primers and Cy3 labeling, and the obtained double-stranded DNAs (targets) were finally hybridized with DNA microarrays (probes). A fluorescent spot on the microarray, detected by scanning indicated positive hybridization, i.e. the involved subtype. The assay was specific as various heterologous viruses or HPAIVs of other subtypes tested were negative. No cross-hybridization among different subtypes could be detected. The assay was more sensitive than RT-PCR and chicken embryo inoculation and could be also used for field samples. Summing up, the assay has proved useful for simultaneous detection and differentiation of main epidemic HPAIV subtypes.

Stable tooth contacts in intercuspal occlusion makes for utilities of the jaw elevators during maximal voluntary clenching.

Data are inconsistent concerning whether the level of the surface electromyographic (SEMG) activity of jaw-closing muscles increases when biting forces elevated during maximal voluntary clenching (MVC). In this study, T-Scan III system and BioEMG III system were used to record bite force, occlusal contacts and SEMG activity of the anterior temporalis (TA) and of the masseter muscles (MM) simultaneously. Recordings were obtained from 16 healthy young adult males during different conditions: (i) a fast MVC from resting position to intercuspal position (ICP); (ii) mandibular movements from ICP to protrusive or lateral edge-to-edge positions with teeth in contact with biting; (iii) a fast MVC in protrusive and lateral edge-to-edge positions. A higher level of SEMG activity was associated with a higher bite force during occluding movements (P < 0.05). However, during fast MVC from rest to ICP, the largest number of occlusal contacts was achieved and distributed more symmetrically, the highest level of biting force was obtained, but the SEMG activity of the jaw elevator muscles was reduced compared with its maximum level (P < 0.05). This phenomenon was not observed during the fast MVC in protrusive or lateral edge-to-edge positions. The present results that a lower SEMG activity was associated with the largest number of occlusal contacts and the highest level of bite force during centric MVC demonstrated a complex integration of jaw-closing muscles when a stable occlusion is present.

Partial cellular reprogramming stably restores the stemness of senescent epidermal stem cells.

OBJECTIVE: Adult stem cell senescence and exhaustion are important drivers of organismal age. Restored stem cell self-renewal has revealed novel therapeutic targets for decreasing the incidence of age-associated diseases (AADs) and prolonging the human health span. Transient ectopic expression of the reprogramming factors Oct3/4, Sox2, Klf4 and c-Myc (collectively known as OSKM) in somatic cells can induce partial cellular reprogramming and effectively ameliorate their age-associated hallmarks. However, how this form of rejuvenation is applied to senescent stem cells remains unknown. MATERIALS AND METHODS: The Integrin-α6highCD71high epidermal stem cells (ESCs) with low self-renewal ability were sorted by flow cytometry and then treated by the interrupted reprogramming induced by transient expression of OSKM. The ability of secondary clones' generation and self-proliferation in vitro, as well as stem cell marker p63, were detected to determine their self-renewal ability. Besides, gene and protein of epidermal cell markers were detected to determine whether their cell identities were retained. Finally, DNA methylation age (eAge) and DNA dehydroxymethylase/methyltransferase were analyzed to explore the alternation of their global DNA methylation pattern during this rejuvenation. RESULTS: The partial reprogramming restored the youthful self-renewal and proliferation in senescent ESCs, including larger secondary clone generation, higher expression of stem cell marker p63 and proliferation marker Ki67, and faster proliferation speed, in each case without abolishing epithelial cellular identity. Moreover, the rejuvenation of adult stem cells could be maintained for 2 weeks after reprogramming factor withdrawal, which was more stable than that of differentiated somatic cells. Additionally, we found that partial reprogramming counteracted the acceleration of eAge in senescent epidermal stem cells and DNA methyltransferase 1 (DNMT1) may play a crucial role in this process. CONCLUSIONS: Partial reprogramming has high therapeutic potential for reversing adult stem cell age, providing an advanced way to treat AADs.

MicroRNA-23b promotes tolerogenic properties of dendritic cells in vitro through inhibiting Notch1/NF-κB signalling pathways.

BACKGROUND: MicroRNAs (miRNAs) are known to regulate the inflammatory response in various cell types. However, the ability of miRNAs to modulate dendritic cells (DCs) function for allergen immunotherapy is unclear. OBJECTIVE: To assess the role of miR-23b in the regulation of ovalbumin (OVA)-induced DC differentiation and function and to investigate the related molecular mechanisms. METHODS: Bone marrow-derived dendritic cells (BMDCs) were generated from murine bone marrow progenitor cells and subsequently stimulated with OVA to examine the profile of miRNA expression. After transfection with miR-23b reagents, DCs were evaluated for endocytic ability, surface marker expression, cytokine secretion and CD4+ T-cell differentiation. The possible roles of the Notch and NF-κB signalling pathways were also evaluated. Human monocyte-derived dendritic cells (MDDCs) were similarly evaluated as well. RESULTS: Significant upregulation of miR-23b was observed in BMDCs pulsed with OVA. Following miR-23b transfection, BMDCs showed decreased OVA uptake, increased IL-10 production, decreased IL-12 production and an enhanced capacity to promote FoxP3+ CD4+ T regulatory cells (Tregs) differentiation. In addition, inactivation of the Notch1 and NF-κB signalling pathways were observed. Conversely, inhibition of miR-23b in BMDCs resulted in the opposite effects. In human MDDCs, miRNA23b transfection similarly increased IL-10 and decreased IL-12 production, and that treated human MDDCs induced increased FoxP3+ CD4+ T cells. CONCLUSION: Our findings provide evidence that miR-23b is capable of inducing tolerogenic DC activity and Treg responses in vitro through the inhibition of the Notch1 and NF-κB signalling pathways; thus, miR-23b might represent a therapeutic target for the management of allergic diseases.

Effects of male fecundity, interindividual distance and anisotropic pollen dispersal on mating success in a Scots pine (Pinus sylvestris) seed orchard.

Quantifying the effect of pollen dispersal and flowering traits on mating success is essential for understanding evolutionary responses to changing environments and establishing strategies for forest tree breeding. This study examined, quantitatively, the effects of male fecundity, interindividual distance and anisotropic pollen dispersal on the mating success of Scots pine (Pinus sylvestris), utilizing a well-mapped Scots pine seed orchard. Paternity analysis of 1021 seeds sampled from 87 trees representing 28 clones showed that 53% of the seeds had at least one potential pollen parent within the orchard. Pronounced variation in paternal contribution was observed among clones. Variations in pollen production explained up to 78% of the variation in mating success, which was 11.2 times greater for clones producing the largest amount of pollen than for clones producing the least pollen. Mating success also varied with intertree distance and direction, which explained up to 28% of the variance. Fertilization between neighboring trees 2.3 m apart was 2.4 times more frequent than between trees 4.6 m apart, and up to 12.4 times higher for trees downwind of the presumed prevailing wind direction than for upwind trees. The effective number of pollen donors recorded in the seed orchard (12.2) was smaller than the theoretical expectation (19.7). Based on the empirical observations, a mating model that best describes the gene dispersal pattern in clonal seed orchards was constructed.

Silica dust, diesel exhaust, and painting work are the significant occupational risk factors for lung cancer in nonsmoking Chinese men.

BACKGROUND: Few epidemiological studies have explored the associations between occupational exposures and lung cancer in lifelong nonsmoking men. METHODS: We obtained lifetime occupational history and other relevant information for 132 newly diagnosed lung cancer cases among nonsmoking Chinese men and 536 nonsmoking community referents. Unconditional multiple logistic regression analysis was performed to estimate the odds ratio (OR) of lung cancer for specific occupational exposures. RESULTS: Significantly increased lung cancer risk was found for nonsmoking workers occupationally exposed to silica dust (OR=2.58, 95% confidence interval (CI): 1.11, 6.01), diesel exhaust (OR=3.47, 95% CI: 1.08, 11.14), spray painting (OR=2.81, 95% CI: 1.14, 6.93), and nonspray painting work (OR=2.36, 95% CI: 1.04, 5.37). Silica dust exposure was associated with a significantly increased risk of adenocarcinoma (OR=2.91, 95% CI: 1.10, 7.68). We observed a positive gradient of all lung cancers and of adenocarcinoma with duration of employment for workers exposed to silica dust and spray painting. CONCLUSION: This study found an increased risk of lung cancer among nonsmoking Chinese men occupationally exposed to silica dust, diesel exhaust, and painting work.

MiR-296-3p promotes the development and progression of preeclampsia via targeting the CEMIP.

OBJECTIVE: Preeclampsia (PE) is one of common pregnancy diseases, which has seriously threatened the health of the gravidas. Although upregulated miR-269-3p has been found in the placentas of the patients with PE, the regulation mechanisms of miR-296-3p remain unclear. PATIENTS AND METHODS: In this study, the placentas of the patients and normal gravidas were used to observe the difference in miR-296-3p expression level, and HTR-8/Svneo and JAR cells were used to investigate the role of miR296-3p in trophoblast cells. Besides, qRT-PCR, Western blot, CCK-8 assay, Dual-Luciferase reporter gene assay and transwell assay were used to explore the functions and regulation mechanisms of miR-296-3p on PE. RESULTS: The results showed that miR-296-3p was upregulated in the PE-placentas, and increased miR-296-3p could inhibit the proliferation, invasion and migration of HTR-8/Svneo and JAR cells. Besides, miR-296-3p could directly target the 3'-UTR of CEMIP, and the phenomena induced by increased miR-296-3p, including decreased ß-catenin and p-AKT and weakened proliferation, invasion and migration abilities, could be reversed by upregulating the expression level of CEMIP. CONCLUSIONS: To summarize, this study suggests that miR-296-3p inactivates the Wnt/ß-catenin and PI3K/AKT pathways to promote the progression of PE via targeting the CEMIP.

Ivy Sign in Moyamoya Disease: A Comparative Study of the FLAIR Vascular Hyperintensity Sign Against Contrast-Enhanced MRI.

BACKGROUND AND PURPOSE: The ability of the ivy sign on contrast-enhanced T1-weighted MR imaging (CEMR) to reflect cerebral perfusion and postoperative revascularization in Moyamoya disease remains largely unknown. We aimed to compare the capabilities of CEMR and FLAIR. MATERIALS AND METHODS: CEMR, FLAIR, arterial spin-labeling, and DSA were performed in 44 patients with Moyamoya disease. The ivy sign was scored separately on CEMR and FLAIR using the Alberta Stroke Program Early CT Score. The status of leptomeningeal collaterals was scored on DSA. The postoperative Matsushima grade was evaluated at least 3 months after surgical revascularization. RESULTS: Scoring of the ivy sign on CEMR showed excellent interrater reliability, and FLAIR vascular hyperintensity showed moderate interrater reliability. Correlation analyses revealed that DSA scores were more consistent with the CEMR-based ivy sign score (r = 0.25, P = .03) than with FLAIR vascular hyperintensity (r = 0.05, P = .65). The CEMR-based ivy sign score was significantly correlated with CBF in late-Suzuki stage Moyamoya disease (t = -2.64, P = .02). The CEMR-based ivy sign score at baseline was significantly correlated with the postoperative Matsushima grade (r = 0.48, P = .03). CONCLUSIONS: In this study, CEMR outperformed FLAIR in capturing the ivy sign in Moyamoya disease. In addition, the CEMR-based ivy sign score provided adequate information on hemodynamic status and postoperative neovascularization. The current study suggested that CEMR could be considered as an alternative to FLAIR in future studies investigating leptomeningeal collaterals in Moyamoya disease.

Time course of action and pharmacokinetics of ropivacaine in adult and elderly patients following combined lumbar plexus-sciatic nerve block.

OBJECTIVE: Large doses of local anesthetics are needed in combined lumbar plexus-sciatic nerve block. The safety of local anesthetics is important among elderly patients. This study investigates the pharmacokinetics of ropivacaine in elderly patients with combined lumbar plexus-sciatic nerve block. METHODS: Ropivacaine pharmacokinetics and the time of onset, duration of sensory and motor block were studied in adult patients (18 - 60 y) and elderly patients (>= 60 y). Combined lumbar plexus-sciatic nerve block was performed using a nerve stimulator. 0.375% ropivacaine was injected in patients. Plasma concentration of ropivacaine was determined by high performance liquid chromatography. RESULTS: There was no significant difference in onset time and time to peak (tmax1, tmax2) between elderly patients and adult patients. Half-life time (t1/2alpha, t1/2beta) and the duration of sensory and motor block were longer in elderly patients (p < 0.05). While the peak plasma concentrations of (double peaks, Cmax1, Cmax2) wer lower in elderly patients than that in adult patients. CONCLUSIONS: Age does not affect the onset time of ropivacaine after combined lumbar plexus-sciatic nerve block but can influence the motor and sensory recovery. Local absorption of ropivacaine is slower in elderly patients compared to adults.

Expression levels of plasma miRNA-21 and NT-proBNP in children with Kawasaki disease and their clinical significance.

OBJECTIVE: The purpose of this study was to detect the expression levels of plasma microRNA-21 (miRNA-21) and NT-proBNP (N-terminal prohormone of brain natriuretic peptide) in children with Kawasaki disease (KD), as well as their clinical significance. PATIENTS AND METHODS: Children with KD (n=100) who were treated in our hospital from June 2017 to May 2019 were included. In the same period, non-KD children with febrile diseases were included as controls. Plasma levels of miRNA-21 and NT-proBNP were detected by reverse transcriptase-polymerase chain reaction (RT-PCR) and electrochemiluminescence, respectively. Then, the relationship between miRNA-21 and NT-proBNP in children with KD was analyzed by Pearson's correlation test. Potential factors influencing KD were analyzed by Multivariate logistic regression test. Finally, receiver operating characteristic (ROC) curves were depicted to assess the diagnostic potentials of miRNA-21 and NT-proBNP in KD. RESULTS: The results showed that miRNA-21 and NT-proBNP levels were higher in children with KD. Plasma level of miRNA-21 was positively correlated with NT-proBNP level in children with KD. Besides, both miRNA-21 and NT-proBNP were risk factors influencing the onset of KD. According to ROC curves, the sensitivity and specificity of miRNA-21 in diagnosing KD was 83% and 89%, respectively (AUC=0.9212, 95% CI: 0.8809-0.9614, cut-off value=1.985). NT-proBNP also displayed diagnostic potential in KD (AUC=0.9788, 95% CI: 0.9630-0.9946, cut-off value=265.6, sensitivity=88%, specificity=95%). CONCLUSIONS: MiRNA-21 and NT-proBNP are upregulated in plasma of children with KD. They are positively correlated with each other and serve as risk factors for KD. Both of them can be utilized as indicators of auxiliary diagnosis in KD.

The roles of smoking and cooking emissions in lung cancer risk among Chinese women in Hong Kong.

BACKGROUND: We conducted this case-control study to evaluate smoking effect on lung cancer conditional on the level of exposure to cooking emissions and to explore whether there is a joint effect of these two risk factors. SUBJECTS AND METHODS: We selected 279 newly diagnosed primary lung cancer cases and 322 community controls from Hong Kong females, frequency matched by age group, and collected relevant data. We applied logistic regression to estimate lung cancer risk related to smoking and cooking fume exposure, expressed as total cooking dish-years, while adjusting for various potential confounding factors. RESULTS: Current smoking was associated with four-fold increased risk, and ex-smoking with two-fold risk, which was not much influenced by cooking dish-years. No increased risk was observed in environmental tobacco smoking. Increasing intakes of yellow/orange vegetables and multivitamins were significant protective factors in all models. In the analysis of joint effect, the combination of smoking and cooking dish-years tended to have a greater risk than exposure to cooking fumes alone. There was a dose-response gradient with total dish-years in nonsmokers, but not in smokers. Smoking was more strongly associated with nonadenocarcinoma, whereas exposure to cooking fumes appeared to be related to both adenocarcinoma and nonadenocarcinoma. CONCLUSION: We confirmed the important roles of smoking and cooking emissions in lung cancer risk among the women. These two major risk factors appeared to act independently.

LncRNA LACAT1 promotes proliferation of oral squamous cell carcinoma cells by inhibiting microRNA-4301.

OBJECTIVE: To investigate the expression of lncRNA LACAT1 in oral squamous cell carcinoma (OSCC) tissues, and to further investigate its potential mechanism in OSCC as well as its relationship with clinicopathology and prognosis. PATIENTS AND METHODS: Quantitative Real-time polymerase chain reaction (qRT-PCR) was used to analyze LACAT1 level in 78 pairs of OSCC tumor tissues and adjacent tissues, and the interplay between LACAT1 level and OSCC clinical indices along with patient's prognosis was analyzed. Further, the expression level of LACAT1 in OSCC cell line was verified by qRT-PCR. In addition, after LACAT1 knockdown model was constructed using lentivirus and transfected into OSCC cell lines, cell cloning assay and flow cytometry were used to analyze the impact of LACAT1 on biological functions of OSCC cells, and finally its association with microRNA-4301 was explored. RESULTS: In this experiment, qRT-PCR results revealed that LACAT1 level in OSCC tumor tissue specimens was significantly higher than that in paracancerous tissues. In addition, the pathology stage in patients with higher level of LACAT1 was also higher while the overall survival rate was lower. Compared with sh-NC group, the cell proliferation ability of sh-LACAT1 group was significantly decreased while cell apoptosis was oppositely increased. QRT-PCR results showed that microRNA-4301 level was significantly elevated in cells of sh-LACAT1 group, suggesting that the expression of above two molecules was negatively correlated. Meanwhile, cell reverse experiment also demonstrated that LACAT1 and microRNA-4301 can be mutually regulated, and thereby promote the malignant progression of OSCC. CONCLUSIONS: The LACAT1 level was found significantly increased in OSCC tissues and cells, which resulted in an advanced OSCC pathological stage and a poor prognosis of patients. In addition, it was found that LACAT1 may promote the malignant progression of OSCC by modulating microRNA-4301 activity.

The regulatory role of SLP-2 and mechanism on CCBE1 gene expression in rectal carcinoma and adjacent lymphatic tube tissues.

OBJECTIVE: The incidence of rectal carcinoma (RC) has been increasing recently, and becomes the second most common digestive tumors besides gastric cancer, with a rise in the incidence of RC in younger populations. The early diagnosis and treatment are thus critical for the improvement of survival rate and life quality of patients. Stomatin-like protein 2 (SLP-2) is a type of membrane factor, which is generally found highly expressed in various tumors. Collagen and calcium-binding EGF domain (CCBE1) belongs to lymphatic tube genesis factor. The regulatory role of SLP-2 gene on CCBE1 expression in RC tumor and adjacent lymphatic tube tissues, however, has not been studied. PATIENTS AND METHODS: 52 RC patients were recruited, and tumor and adjacent lymphatic tube tissues were collected. Real-time PCR, western blotting and immunohistochemistry (IHC) staining were used to analyze SLP-2 and CCBE1 expressions. Human lymphatic endothelial cells (LECs) were cultured in vitro and were assigned to control, scramble, and SLP-2 siRNA group. MTT assay was used to detect cell proliferation, while caspase 3 activity was detected. RESULTS: SLP-2 and CCBE1 levels were significantly elevated in tumor lymphatic tissues, compared to that in adjacent tissues. Statistically positive correlation between SLP-1 and CCBE2 was found (p<0.05). The downregulation of SLP-2 by siRNA inhibited cell proliferation, elevated caspase3 activity, and decreased CCBE1 expression (p<0.05 compared to control group). CONCLUSIONS: SLP-2 is up-regulated in RC lymphatic tissues, and is positively correlated with the level of CCBE1, which provides the academic the basis for the development of medicine targeting SLP-2 in the anti-rectal carcinoma therapy.

Association between downexpression of miR-1301 and poor prognosis in patients with glioma.

OBJECTIVE: MiR-1301 has been shown to be frequently down-regulated in various tumors. However, the clinical significance of miR-1301 in human glioma is still unclear. The aim of this study was to evaluate the prognostic impact of the expressions of miR-1301 in patients with glioma. PATIENTS AND METHODS: Quantitative Real-time PCR was used to determine the expression miR-1301 in glioma tissues and pair-matched adjacent normal tissues. The relationships between miR-1301 expression and clinicopathological parameters were examined by X2 test. Kaplan-Meier curves and multivariate Cox proportional models were used to study the impact on clinical outcome. RESULTS: We observed that miR-1301 expression was significantly lower in glioma tissues compared with adjacent non-cancerous tissues (p<0.01). Also, low expressions of miR-1301 were significantly associated with high WHO grade (p<0.006), low Karnofsky performance score (KPS) (p=0.001), and large tumor size (p=0.004). Furthermore, the data of Kaplan-Meier survival analysis showed that low miR-1301 expression significantly associated with a worse overall survival (p=0.003) and disease-specific survival (p=0.001). Finally, the univariate and multivariate analysis showed that the miR-1301 expression was an independent predictor for both overall survival and disease-specific survival in glioma. CONCLUSIONS: Our findings suggested lower miR-1301 expression resulted in poorer survival in patients with glioma, which may provide important indicators for further research.

MiR-616 promotes proliferation and inhibits apoptosis in glioma cells by suppressing expression of SOX7 via the Wnt signaling pathway.

OBJECTIVE: Accumulating evidence has indicated that miR-616 exerts tumor promoter roles in several types of cancer. However, the expression pattern and roles of miR-616 in glioma progression remain unknown. This study aimed to reveal the role of miR-616 in glioma cell proliferation and its potential mechanisms. PATIENTS AND METHODS: Real-time polymerase chain reaction was used to assay the expression of miR-616 in glioma tissue samples and glioma cell lines. MTT proliferation assay and flow cytometry analysis were performed to test the apoptosis and proliferation of glioma cell after down-regulation of miR-616. The target of miR-616 was predicted by TargetScan and confirmed by luciferase reporter assay. Changes in Wnt signaling markers expression were assessed using Western blotting. RESULTS: We found that the expression of miR-616 was increased in glioma tissues and cell lines. MTT and low cytometry analysis indicated that down-regulation of miR-616 significantly inhibited proliferation and promoted apoptosis in glioma cells. Moreover, SOX7 was confirmed to be a direct target of miR-616 in glioma cells using luciferase assay and Western blotting. Finally, it was found that down-regulation of miR-616 or upregulation of SOX7 could suppress the activity of Wnt/ß-catenin signaling in glioma cells. CONCLUSIONS: Our findings indicated that miR-616 acted as a tumor promoter in glioma, and its oncogenic roles were involved in the regulation of SOX7 and Wnt/ß-catenin signaling. Moreover, knockdown of miR-616 may provide a potential therapeutic strategy for glioma.

MicroRNA-202 induces cell cycle arrest and apoptosis in lung cancer cells through targeting cyclin D1.

OBJECTIVE: MicroRNAs are critical regulators in tumorigenesis. This study is aimed at investigating the function of miR-202 in the proliferation of human lung cancer cells. PATIENTS AND METHODS: Lung cancer tissues and paired adjacent normal tissues were collected from 20 patients; the expression of miR-202 was tested by Realtime PCR; cell proliferation and cell cycle distribution were determined by CCK-8 assay and flow cytometry, respectively; apoptosis was determined by TUNEL assay and Western blot analysis of cleaved-PARP; the relationship between miR-202 and cyclin D1 mRNA 3'UTR was determined by luciferase activity assay. RESULTS: MiR-202 was significantly reduced in lung cancer tissues compared with the adjacent normal tissues. Overexpression of miR-202 inhibited cell proliferation and induced a G0/G1 cell cycle arrest and apoptosis. Luciferase activity assay and Western blot analysis together showed that miR-202 can bind to the 3'UTR of cyclin D1 mRNA directly and inhibits cyclin D1 expression at the protein level. In addition, restoration of cyclin D1 expression partially abolished cell cycle arrest and apoptosis induced by miR-202. CONCLUSIONS: MiR-202 is constantly downregulated in lung cancer and functions as a tumor suppressive gene via targeting cyclin D1. Modulating the level of miR-202 may be a novel therapeutic method for lung cancer.