Inhibition of A2AR gene methylation alleviates white matter lesions in chronic cerebral hypoperfusion rats.

OBJECTIVE: Chronic cerebral hypoperfusion (CCH) can cause ischemic cerebral white matter lesions (IWML). The aim of this study was to explore the roles of A2A receptors (A2AR) in IWML and the effect of methylation in A2AR gene. MATERIALS AND METHODS: SD rat model of CCH was constructed by bilateral common carotid artery occlusion (BCCAO) method. The rats were then treated with DNA methyltransferase (DNMT) inhibitor (decitabine), agonist (CGS21680) and A2AR inhibitor (SCH58261). Morris water maze and Kluver-Barrera staining were used to assess spatial learning and reference memory after IWML, respectively. Gene transcription and protein expression were measured by qRT-PCR, Enzyme-linked immunosorbent assay (ELISA) and Western blotting, respectively. The concentration of malondialdehyde (MDA), activity of superoxide dismutase (SOD) and DNMT were detected by assay kit. Methylation of A2AR gene promoter region was detected by bisulfite sequencing PCR (BSP). RESULTS: We found that the down-regulated expression of A2AR in corpus callosum under CCH was associated with IWML and cognitive impairment. We further showed that A2AR agonist can reduce the IWML under CCH, and A2AR inhibitor can aggravate the IWML under CCH. We also found that the expression level of DNMTs in corpus callosum and the methylation level in the promoter region of A2AR gene were increased under CCH. DNMT inhibitors could protect white matter by inhibiting the methylation of A2AR promoter and rescue the downregulation of A2AR under CCH. CONCLUSIONS: Our results demonstrate that the downregulation of A2AR mediates IWML in CCH, and A2AR downregulation is related to the increased methylation of A2AR gene promoter. DNMT inhibitors play a protective role in IWML.

MiRNA-488-3p inhibits malignant progression of NSCLC by modulating ADAM9.

Since this article has been suspected of research misconduct and the corresponding authors did not respond to our request to prove originality of data and figures, "MiRNA-488-3p inhibits malignant progression of NSCLC by modulating ADAM9, by Y. Wu, Y. Wu, X.-Y. Chen, Y.-X. Niu, F.-Z. Lv, W. Gao, published in Eur Rev Med Pharmacol Sci 2020; 24 (17): 8893-8901-DOI: 10.26355/eurrev_202009_22830-PMID: 32964979" has been withdrawn. The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/22830.

LINC00665 facilitates the progression of osteosarcoma via sponging miR-3619-5p.

OBJECTIVE: Long non-coding RNAs (lncRNAs) play vital roles in the pathogenesis and development of multiple cancers, including osteosarcoma (OS). The present study aims to investigate the role of LINC00665 in OS progression. PATIENTS AND METHODS: The expression levels of LINC00665 and miR-3619 were assessed by RT-qPCR. The correlation between LINC00665 and miR-3619 expression was evaluated by Pearson's correlation analysis. The interaction between LINC00665 and miR-3619 was predicted by starBase, which was further confirmed by Luciferase reporter assay and RIP assay. The viability, invasion, and migration of OS cells were analyzed by CCK-8 and transwell assays. RESULTS: LINC00665 expression was upregulated in OS tissues and cell lines, and the high level of LINC00665 was associated with poor prognosis in OS. Moreover, LINC00665 knockdown attenuated the viability, invasion, and migration of OS cells. In addition, miR-3619 was demonstrated to be a target of LINC00665. Overexpression of miR-3619 inhibited OS progression, while this effect was abolished by the upregulation of LINC00665. CONCLUSIONS: We demonstrated that LINC 00665 accelerated OS development by targeting miR-3619. These findings might provide potential treatment strategies for patients with OS.

MiRNA-488-3p inhibits malignant progression of NSCLC by modulating ADAM9.

OBJECTIVE: The purpose of this study was to investigate the role of microRNA-488-3p in the proliferation, invasion and migration of lung cancer cells and to further explore the potential regulatory mechanisms. PATIENTS AND METHODS: MicroRNA-488-3p expression in 46 pairs of tumor tissue and paracancerous tissue specimens collected from non-small cell lung cancer (NSCLC) patients were measured through quantitative real-time polymerase chain reaction (qRT-PCR) method, and the interplay between microRNA-488-3p expression and some clinical indicators of these subjects was also analyzed. In addition, microRNA-488-3p overexpression models were constructed in NSCLC cell lines, and then Cell Counting Kit-8 (CCK-8) test and transwell assays were carried out to evaluate the effect of microRNA-488-3p on the NSCLC cell functions. Furthermore, bioinformatics analysis and luciferase reporter gene assay were carried out to uncover the potential interaction between microRNA-488-3p and its downstream gene ADAM9. RESULTS: QPCR results revealed that microRNA-488-3p showed a significant lower expression in NSCLC tissue samples than in adjacent normal ones. In comparison to patients with high expression of microRNA-488-3, patients with low expression of microRNA-488-3 exhibited higher incidence of lymph node or distant metastasis and lower survival rate. In vitro cell experiments showed that, in comparison to control group, overexpression of microRNA-488-3p significantly weakened the proliferation ability as well as the invasion and migration of NSCLC cells. Subsequently, a significant increase in ADAM9 expression in NSCLC tissue samples was found, which indicated its negative correlation with microRNA-488-3p. In addition, cell recovery experiment demonstrated that overexpression of ADAM9 could counteract the impact of microRNA-488-3p upregulation on the proliferation and invasion ability of NSCLC cells, and the two may thus together affect the malignant progression of NSCLC. CONCLUSIONS: It can be concluded that microRNA-488-3p, which is associated with the incidence of metastasis in NSCLC patients, can inhibit the malignant progression of NSCLC cells by modulating ADAM9 expression.

LncNORAD interference inhibits tumor growth and lung cancer cell proliferation, invasion and migration by down-regulating CXCR4 to suppress RhoA/ROCK signaling pathway.

OBJECTIVE: Non-small cell lung cancer (NSCLC) accounts for the majority of lung cancer, with an unfavorable prognosis of 5-year survival rates. It is of great clinical significance to further search for more efficacious and novel targets for diagnosis and therapeutic strategies. This study aimed at clarifying the role of long non-coding RNA (lncRNA) NORAD in proliferation, invasion and migration and tumor growth of NSCLC. MATERIALS AND METHODS: In this study, mRNA levels of lncRNA NORAD were examined by RT-PCR. CCK-8 assay was applied to test cell viability. Furthermore, wound healing assay and transwell assay were performed to detect the migration and invasion of A549 cells, respectively. Immunohistochemistry was applied to assess the levels of CXC chemokine receptor (CXCR) 4 and CXC chemokine ligand (CXCL) 12. Mice models of NSCLC in vivo were exploited to further examine the potential role of NORAD in tumor growth. Key proteins related to Ras homolog gene family member A (RhoA) GTPase/Rho-associated kinase (RhoA/ROCK) pathway were determined by Western blot. RESULTS: NORAD has elevated the levels in NSCLC tissues and cells. NORAD interference dramatically inhibited tumor growth and suppressed A549 cell proliferation, migration and invasion by downregulating CXCR4 and CXCL12 expression. RhoA/ROCK signaling pathway was activated in NSCLC. CONCLUSIONS: This study revealed that the downregulation of lncRNA NORAD could slow down the progression of NSCLC by regulating CXCR4 and RhoA/ROCK signaling pathway.

Treatment and recurrence on re-treatment tuberculosis patients: a randomized clinical trial and 7-year perspective cohort study in China.

Previously treated TB patients still pose a serious threat to global control of TB, yet new re-treatment therapies were little studied. This study aimed to examine the therapeutic effects of new re-treatment regimens, and explore risk factors associated with recurrence after successful treatment. We conducted a cohort study in nine regions of China and enrolled previously treated TB patients from October, 2008 to December, 2010. Patients were randomly divided into four treatment regimen groups including standard, high-dose, long-course, and individualized treatment. After treatment, those with successful treatment outcomes were followed up to 7 years. The effects of different regimens and the information of recurrence were recorded. Risk factors to poor treatment outcomes were calculated using logistic regression model, while risk factors to recurrence or death were calculated using Cox model. Four hundred ninety-two participants were enrolled during the study time and 419 patients were included in our analysis of treatment effects. Overall, the treatment success rate is 75.9%, and the recurrence and death rate is 6.9% and 3.8%, respectively. Reduced risks of poor outcomes were observed in patients who were treated with high-dose and individualized regimen compared with standard regimen, and the adjusted ORs were 0.3 (0.1-0.6), 0.2 (0.1-0.5), respectively. In our analysis of factors associated with recurrence, all documented variables were not significant. Revised re-treatment regimen has better therapeutic effects compared to standard regimen, but it was not associated with lower risk of TB recurrence. Further studies are warranted to evaluate the role of other revised re-treatment regimens in recurrence risk. Trial registration: chictr.org Identifier: ChiCTR1800017441.

ARHGAP15 regulates lung cancer cell proliferation and metastasis via the STAT3 pathway.

OBJECTIVE: Lung cancer, which is typically diagnosed at later stages, is a leading cause of cancer death among both males and females given its highly invasive and rapidly metastasizing nature. Rho GTPase activating protein 15 (ARHGAP15) is a member of the RhoGAP family and functions in multiple biological processes, such as cell proliferation and migration. However, the effect of ARHGAP15 in lung cancer and its underlying molecular mechanisms remain unclear. PATIENTS AND METHODS: In this study, immunohistochemistry and Real Time PCR were performed to detect ARHGAP15 expression in lung cancer tissues and cells. Proliferation, transwell, and Western blot assays were further performed to explore the role and underlying mechanism of ARHGAP15 in lung cancer. RESULTS: Reduced ARHGAP15 expression was observed in lung cancer tissues and cells. In vitro upregulation of ARHGAP15 in lung cancer cells strongly suppressed cell proliferation, migration, and invasion and was accompanied by reduced matrix metalloproteinase-2 (MMP2), MMP9, vascular endothelial growth factor (VEGF) expression, and the phosphorylation of the signal transducer and activator of transcription-3 (p-STAT3). In contrast, interleukin-6 (IL-6) had the opposite effect and the induction of IL-6 was counteracted by ARHGAP15 upregulation. In addition, the proliferation, migration, and cell invasion induced by ARHGAP15 silencing were potentially inhibited by the STAT3 inhibitor AG490 (100 µM), MMP2, MMP9, VEGF, and p-STAT3 levels decreased. CONCLUSIONS: These results suggest that ARGFAP15 suppressed the proliferation and metastasis of lung cancer cells, which may occur through the inhibition of MMP2, MMP9, and VEGF expression via the STAT3 pathway inactivation.

Correlation study on ß2-adrenergic receptor gene polymorphisms and asthma susceptibility: evidence based on 57 case-control studies.

OBJECTIVE: Previous studies have indicated that ß2-adrenergic receptor (ADRB2) genetic polymorphism is related to the risk of asthma, but the results still have some controversy and uncertainty. To this end, the meta-analysis was performed, including all studies that can be used to assess the correlation between ADRB2 polymorphism and asthma susceptibility. MATERIALS AND METHODS: The related papers on ADRB2 polymorphisms and asthma were systematically reviewed in databases of PubMed, EMBASE, Cochrane Library, and WanFang, Odds ratios (ORs) and corresponding 95% confidence intervals (95% CIs) were measured. The sensitivity analysis and publication bias were evaluated to investigate the correlation. RESULTS: This meta-analysis included 57 papers in total involving 11,157 cases and 12,281 controls. Results illustrated that the C79G variant genotypes owned a reduced effect on asthma susceptibility (G vs. C: OR=0.94, p=0.037). In the age stratification analysis, C79G polymorphism owned a reduced effect on asthma risk for children (GG vs. CC: OR=0.69, p=0.002; GG vs. CC+CG: OR=0.65, p<0.001). Furthermore, in the ethnic stratification analysis, the C79G variant genotypes also owned a reduced effect on asthma in Asians (GG vs. CC: OR=0.80, p=0.027; GG vs. CC+CG: OR=0.81, p=0.02). Besides, for A46G polymorphism, the ethnic stratification analysis demonstrated that the A46G variant owned an increased effect on asthma susceptibility among Caucasians (G vs. A: OR=1.15, p=0.043). For C491T polymorphism, a considerable reduced effect was found between C491T and asthma susceptibility for children (CT vs. CC: OR=0.70, p=0.03). In the ethnic stratification analysis, the effect was also considerable in the Caucasian subjects. CONCLUSIONS: The present meta-analysis demonstrated that C79G and C491T polymorphism may be a defensive factor for asthma, while A46G polymorphism may be a risk factor for asthma among the Caucasian population.

Vitamin D receptor BsmI polymorphism may be associated with an decreased osteoporosis risk in South China.

Although many epidemiology studies have evaluated the correlation between vitamin D receptor (VDR) BsmI polymorphism and osteoporosis, the current results remain inconclusive. This meta-analysis updated and reevaluated the possible association between VDR BsmI polymorphism and osteoporosis in Chinese populations. Studies were identified using PubMed and Chinese databases through December 2016. The associations were assessed with pooled odds ratios (ORs) and 95% confidence intervals (CIs). A total of nine studies with 688 osteoporosis cases and 730 controls were included in this meta-analysis. Overall, no significant association was found between VDR BsmI polymorphism and osteoporosis when all studies in Chinese populations were pooled into this meta-analysis. In subgroup analyses, significantly decreased risk was found in South China (bb vs. BB: OR = 0.22, 95% CI = 0.06-0.76; bb+Bb vs.BB: OR = 0.27, 95% CI = 0.09-0.81). This meta-analysis suggests that VDR BsmI polymorphism may have a protective role against the development of osteoporosis in South China.

Adenosine A2A receptor inactivation alleviates early-onset cognitive dysfunction after traumatic brain injury involving an inhibition of tau hyperphosphorylation.

Tau is a microtubule-associated protein, and the oligomeric and hyperphosphorylated forms of tau are increased significantly after neurotrauma and considered important factors in mediating cognitive dysfunction. Blockade of adenosine A2A receptors, either by caffeine or gene knockout (KO), alleviates cognitive dysfunction after traumatic brain injury (TBI). We postulated that A2AR activation exacerbates cognitive impairment via promoting tau hyperphosphorylation. Using a mouse model of moderate controlled cortical impact, we showed that TBI induced hyperphosphorylated tau (p-tau) in the hippocampal dentate gyrus and spatial memory deficiency in the Morris water maze test at 7 days and 4 weeks after TBI. Importantly, pharmacological blockade (A2AR antagonist ZM241385 or non-selective adenosine receptor antagonist caffeine) or genetic inactivation of A2ARs reduced the level of tau phosphorylation at Ser404 and alleviated spatial memory dysfunction. The A2AR control of p-tau is further supported by the observations that a KO of A2AR decreased the activity of the tau phosphorylation kinases, glycogen synthase kinase-3ß (GSK-3ß) and protein kinase A (PKA) after TBI, and by that CGS21680 (A2AR agonist) exacerbated okadaic acid-induced tau hyperphosphorylation in cultured primary hippocampal neurons. Lastly, CGS21680-induced neuronal tau hyperphosphorylation and axonal injury were effectively alleviated by individual treatments with ZM241385 (A2AR antagonist), H89 (PKA antagonist) and SB216763 (GSK-3ß antagonist), or by the combined treatment with H89 and SB216763. Our findings suggest a novel mechanism whereby A2AR activation triggers cognitive dysfunction by increasing the phosphorylation level of tau protein after TBI and suggest a promising therapeutic and prophylactic strategy by targeting aberrant A2AR signaling via tau phosphorylation.

Entecavir maleate versus entecavir in Chinese chronic hepatitis B predominantly genotype B or C: Results at week 144.

Reports on the efficacy and safety of long-term entecavir treatment in chronic hepatitis B (CHB) predominantly genotype B or C are insufficient. This study presents the efficacy and safety of entecavir maleate in Chinese CHB patients. Patients were randomly assigned to receive 48-week treatment with either 0.5 mg/day entecavir (group A) or 0.5 mg/day entecavir maleate (group B), and then all patients received treatment with 0.5 mg/day entecavir maleate from week 49. Two hundred and seventy-five patients with CHB (HBeAg-positive: 218) were analysed, predominantly (98.5%) with genotype B or C. Baseline characteristics were balanced. For the HBeAg-positive CHB patients, the mean HBV DNA level decreased similarly (A: by 6.36 log10 IU/mL vs B: by 6.31 log10 IU/mL) between groups at week 144. The percentages of patients who achieved undetectable HBV DNA were similar (A: 70.59% vs B: 66.67%) between groups. Similar HBeAg loss rates (A: 43.53% vs B: 40.23%; P>.05) and HBeAg seroconversion rates (A: 21.52% vs B: 21.18%) were achieved. For the HBeAg-negative CHB patients, similar reductions in HBV DNA levels from baseline (A: by 6.13 log10 IU/mL vs B: by 5.65 log10 IU/mL) and percentages of patients who achieved undetectable HBV DNA (A: 100% vs B: 100%) were achieved. The overall incidence of adverse events was comparable between groups. In conclusions, 48-week administration of entecavir maleate and entecavir showed similar efficacy and safety in Chinese patients with CHB. Long-term entecavir maleate treatment was effective and safe in CHB patients.

Baseline hepatitis B core antibody predicts treatment response in chronic hepatitis B patients receiving long-term entecavir.

Studies regarding the clinical significance of quantitative hepatitis B core antibody (anti-HBc) in patients with chronic hepatitis B receiving first-line nucleos(t)ide analogues is limited. The aim of this study was to determine the performance of anti-HBc as a predictor for hepatitis B e antigen (HBeAg) seroconversion in HBeAg-positive CHB patients treated with entecavir. This was a retrospective cohort study consisting of 139 Chinese patients enrolled in a multicenter clinical trial treated with entecavir or entecavir maleate for up to 240 weeks. Anti-HBc evaluation was conducted for all the available samples using a newly developed double-sandwich anti-HBc immunoassay. At week 240, 35 (25.2%) patients achieved a serological response (HBeAg seroconversion) and these patients at week 240 had significantly higher levels of anti-HBc (P<.01). We defined 4.65 log10  IU·mL-1 , with a maximum sum of sensitivity and specificity, as the optimal cut-off value of baseline anti-HBc level to predict seroconversion. Patients with baseline anti-HBc ≥4.65 log10  IU·mL-1 had 28.0% (26/93) and 35.5% (33/93) chance of seroconversion at weeks 144 and 240, respectively. The baseline anti-HBc level was the strongest predictor for seroconversion at week 144 (OR: 5.78, 95% confidence interval [CI]: 2.05-16.34, P=.001). The baseline anti-HBc level was a strong predictor for seroconversion at week 240 (OR: 5.36, 95% CI: 2.17-13.25, P<.001). Hence, baseline anti-HBc titre is a useful predictor of long-term entecavir therapy efficacy in HBeAg-positive CHB patients, which could be used to optimize antiviral therapy.

Investigating ego modules and pathways in osteosarcoma by integrating the EgoNet algorithm and pathway analysis

Osteosarcoma (OS) is the most common primary bone malignancy, but current therapies are far from effective for all patients. A better understanding of the pathological mechanism of OS may help to achieve new treatments for this tumor. Hence, the objective of this study was to investigate ego modules and pathways in OS utilizing EgoNet algorithm and pathway-related analysis, and reveal pathological mechanisms underlying OS. The EgoNet algorithm comprises four steps: constructing background protein-protein interaction (PPI) network (PPIN) based on gene expression data and PPI data; extracting differential expression network (DEN) from the background PPIN; identifying ego genes according to topological features of genes in reweighted DEN; and collecting ego modules using module search by ego gene expansion. Consequently, we obtained 5 ego modules (Modules 2, 3, 4, 5, and 6) in total. After applying the permutation test, all presented statistical significance between OS and normal controls. Finally, pathway enrichment analysis combined with Reactome pathway database was performed to investigate pathways, and Fisher's exact test was conducted to capture ego pathways for OS. The ego pathway for Module 2 was CLEC7A/inflammasome pathway, while for Module 3 a tetrasaccharide linker sequence was required for glycosaminoglycan (GAG) synthesis, and for Module 6 was the Rho GTPase cycle. Interestingly, genes in Modules 4 and 5 were enriched in the same pathway, the 2-LTR circle formation. In conclusion, the ego modules and pathways might be potential biomarkers for OS therapeutic index, and give great insight of the molecular mechanism underlying this tumor.

Resveratrol and STAT inhibitor enhance autophagy in ovarian cancer cells.

Autophagic activity reflects cellular response to drug treatment and can be regulated by STAT3 signaling. Resveratrol inhibits STAT3 activation and causes remarkable growth arrest and cell death of ovarian cancer (OC) cells. However, the autophagic status and its relevance with resveratrol's anti-OC effects remain unclear. We analyzed the states of autophagic activities, the nature of autophagosomes and the levels of autophagy-related proteins (LC-3, Beclin 1 and STAT3) in resveratrol-treated CAOV-3 and OVCAR-3 OC cells using multiple approaches. We elucidated the correlation of STAT3 inhibition with autophagic activity by treating OC cells with an upstream inhibitor of STAT proteins, AG490. Resveratrol efficiently suppressed growth, induced apoptosis and inactivated STAT3 signaling of the two OC cell lines. We found enhanced autophagic activity accompanied with Beclin-1 upregulation and LC3 enzymatic cleavage in resveratrol-treated OC cells. Immunofluorescent (IF) microscopic and IF-based confocal examinations demonstrated the accumulation of cytoplasmic granules co-labeled with LC3 and cytochrome C in resveratrol- or AG490-treated OC cells. Using electron microscopy, we confirmed an increase in autophagosomes and mitochondrial spheroids in either resveratrol- or AG490-treated OC cells. This study demonstrates the abilities of resveratrol to enhance apoptotic and autophagic activities in OC cells, presumably via inactivating STAT3 signaling. Resveratrol or the selective JAK2 inhibitor also leads to mitochondrial turnover, which would be unfavorable for OC cell survival and sensitize OC cells to resveratrol.

Whole genome sequencing identified new somatic mutations for chronic myelomonocytic leukemia.

OBJECTIVE: We aimed to gain new insight into the molecular alterations of Chronic Myelomonocytic Leukemia (CMML). PATIENTS AND METHODS: We performed whole-genome sequencing (WGS) and subsequent Sanger sequencing validation analysis in three individuals with CMML. Genomic DNA samples from bone marrow and matching buccal mucosa samples were sequenced. RESULTS: For all six samples, a total of 806.43 Gb data were generated, achieving a minimum mean depth of 30.76. A total of 22 somatic variants were found to be protein-altering, including 1 exonic frame shift indel, 18 missense SNVs, 2 stop gain SNVs, and 1 stop loss SNV. We focused on the five novel variants which have not been reported in known databases and successfully validated three missense SNVs in AKAP4, COL2A1, and MAML1, respectively. CONCLUSIONS: WGS analyzes provided us a new insight into the molecular events governing the pathogenesis of CMML. The somatic variants we reported here may provide new targets for further therapeutic studies.

Construction of a GeogDetector-based model system to indicate the potential occurrence of grasshoppers in Inner Mongolia steppe habitats.

Grasshopper plagues have seriously disturbed grassland ecosystems in Inner Mongolia, China. The accurate prediction of grasshopper infestations and control of grasshopper plagues have become urgent needs. We sampled 234, 342, 335, and 369 plots in Xianghuangqi County of Xilingol League in 2010, 2011, 2012, and 2013, respectively, and measured the density of the most dominant grasshopper species, Oedaleus decorus asiaticus, and the latitude, longitude, and associated relatively stable habitat factors at each plot. We used Excel-GeogDetector software to explore the effects of individual habitat factors and the two-factor interactions on grasshopper density. We estimated the membership of each grasshopper density rank and determined the weights of each habitat category. These results were used to construct a model system evaluating grasshopper habitat suitability. The results showed that our evaluation system was reliable and the fuzzy evaluation scores of grasshopper habitat suitability were good indicators of potential occurrence of grasshoppers. The effects of the two-factor interactions on grasshopper density were greater than the effects of any individual factors. O. d. asiaticus was most likely to be found at elevations of 1300-1400 m, flat terrain or slopes of 4-6°, typical chestnut soil with 70-80% sand content in the top 5 cm of soil, and medium-coverage grassland. The species preferred temperate bunchgrass steppe dominated by Stipa krylovii and Cleistogenes squarrosa. These findings may be used to improve models to predict grasshopper occurrence and to develop management guidelines to control grasshopper plagues by changing habitats.

PTEN inactivation by germline/somatic c.950_953delTACT mutation in patients with Lhermitte-Duclos disease manifesting progressive phenotypes.

Lhermitte-Duclos disease (LDD), a neurological manifestation of Cowden syndrome (CS), is a rare and benign cerebellar disorder, featured by dysplastic cerebellar ganglion cells which replace granular and Purkinje cells. Phosphatase and Tensin Homolog (PTEN) is confirmed as the susceptibility gene for CS which represents the most complex features and is not easily recognizable. We reported two index patients with LDD diagnosed either in an isolated form or coexist with CS. These two patients displayed progressive though comparable phenotypes and were found to carry an identical PTEN c.950_953delTACT mutation in either germline or somatic sources of DNA, respectively. Negative or moderate expression levels of PTEN were validated by immunohistochemistry in the corresponding patients' affected tissues. This study has revealed a novel pathogenicity locus to LDD/CS as a candidate for early molecular diagnosis. In addition, the differential PTEN mutation status with corresponding LDD phenotypes suggests a potential correlation between germline or somatic mutation and coexisting LDD/CS or isolated LDD, respectively. Furthermore, our data could lend some reference to the underlying molecular mechanism of LDD pathogenesis in the future.

Application of magnifying endoscopy with narrow-band imaging in diagnosing gastric lesions: a prospective study.

BACKGROUND: Magnifying endoscopy with narrow-band imaging (ME-NBI) can more clearly assess the surface pattern and microvascular architecture of gastric lesions. OBJECTIVE: To evaluate the diagnostic efficacy of ME-NBI in patients with early gastric cancer. DESIGN: Prospective study. SETTING: Single academic center. PATIENTS: This study involved 164 suspected gastric lesions in 146 consecutive patients who underwent ME-NBI for additional differential diagnosis before treatment. INTERVENTION: ME-NBI findings were classified into 3 groups based on irregularities, absence of surface pattern, and microvascular architecture. All lesions were treated endoscopically or surgically, and ME-NBI diagnosis was compared with histopathological findings. MAIN OUTCOME MEASUREMENTS: Sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of real-time ME-NBI diagnosis were determined. RESULTS: The sensitivity, specificity, and accuracy of ME-NBI were 97.3%, 84.4%, and 90.2%, respectively, in distinguishing between cancerous and noncancerous lesions and were 92.3%, 89.7%, and 90.4%, respectively, in distinguishing undifferentiated from differentiated adenocarcinoma. ME-NBI accurately predicted depth of invasion in 37 of 39 differentiated adenocarcinomas (95%). LIMITATIONS: The sample size was relatively small. CONCLUSIONS: ME-NBI can successfully distinguish between cancerous and noncancerous lesions and between undifferentiated and differentiated adenocarcinomas. Of the 3 patterns on ME-NBI, type A is mainly characteristic of noncancerous lesions, type B is a good indicator of differentiated adenocarcinoma and intramucosal/superficially invasive cancers, and type C is indicative of undifferentiated adenocarcinoma or differentiated cancer with deep submucosal invasion.

Modulation of the activity of globus pallidus by dopamine D1-like receptors in parkinsonian rats.

Morphological studies have shown that the globus pallidus receives dopaminergic innervation from the collaterals of nigrostriatal fibers. Dopamine D1-like receptors are expressed at both pre- and postsynaptic membrane. In the present study, we investigate the in vivo electrophysiological and behavioral effects of pallidal dopamine D1-like receptors in parkinsonian rats. On the lesioned side of 6-hydroxydopamine (6-OHDA) parkinsonian rats, micropressure ejection of dopamine D1-like receptor agonist, SKF38393, increased (88.2 ± 18.6%) the firing rate in 10 out of the 32 pallidal neurons, but decreased (49.5 ± 6.1%) the firing rate in 14 out of the 32 neurons. Furthermore, on the unlesioned side of parkinsonian rats, SKF38393 increased (43.0 ± 6.3%) the firing rate in 9 out of the 30 pallidal neurons, but decreased (47.1 ± 4.8%) the firing rate in 13 out of the 30 neurons. In behaving rats, unilateral microinjection of SKF38393 led to contralateral deflection in the presence of systemic haloperidol administration. The selective dopamine D1-like receptor antagonist, SCH23390, blocked both SKF38393-induced electrophysiological and behavioral effects. Combining electrophysiological and behavioral findings, we concluded that activation of dopamine D1-like receptors modulates the activity of globus pallidus neurons in rats.

Use of chromosome walking in discovery of single-nucleotide polymorphism in noncoding regions of a candidate actin gene in Pinus radiata.

Untranslated regions (UTRs) of eukaryotic mRNAs play crucial roles in post-transcriptional regulation of gene expression via the modulation of nucleocytoplasmic mRNA transport, translation efficiency, subcellular localization, and message stability. Single-nucleotide polymorphisms (SNPs) in UTRs of a candidate gene may also change the post-transcriptional regulation of a gene or function by nucleotide mutation. For species that have not been entirely sequenced genomically, new methods need to be devised to discover SNPs in noncoding regions of candidate genes. In this study, based on the expressed sequence tag (EST) of Pinus radiata (Monterey pine), we obtained all the sequences of UTRs of the actin gene by using a chromosome walking method. We also detected all the SNPs in and around the coding region of the actin gene. In this way, the full genomic sequence (2154 bp) of the actin gene was identified, including the 5’UTR, introns, the coding sequence, and the 3’UTR. PCR amplification and DNA fragment sequencing from 200 unrelated P. radiata trees revealed a total of 21 SNPs in the actin gene, of which 3 were located in the 5’UTR, 3 in the introns, 10 in the coding sequence, and 5 in the 3’UTR. We show that chromosome walking can be used for obtaining the sequence of UTRs, and then, based on this sequence, to discover SNPs in the noncoding regions of candidate genes from this species without an entire genomic sequence.