Distinct metabolites affect the phloem fungal communities in ash trees (Fraxinus spp.) native and nonnative to the highly invasive emerald ash borer (AGRILUS PLANIPENNIS).

Emerald ash borer (EAB, Agrilus planipennis) is an invasive killer of ash trees (Fraxinus spp.) in North America and Europe. Ash species co-evolved with EAB in their native range in Asia are mostly resistant, although the precise mechanism(s) remain unclear. Very little is also known about EAB or ash tree microbiomes. We performed the first joint comparison of phloem mycobiome and metabolites between a native and a nonnative ash species, infested and uninfested with EAB, in conjunction with investigation of larval mycobiome. Phloem mycobiome communities differed between the tree species, but both were unaffected by EAB infestation. Several indicator taxa in the larval gut shared a similarly high relative abundance only with the native host trees. Widely targeted metabolomics revealed 24 distinct metabolites in native trees and 53 metabolites in nonnative trees, respectively, that differed in relative content between infested and uninfested trees only in one species. Interestingly, four metabolites shared a strong relationship with the phloem mycobiomes, majority of which affected only the native trees. Collectively, our results demonstrate a complex interplay between host tree chemistry and mycobiome, and suggest the shared relationships between the mycobiomes of the native host tree and EAB may reflect their shared co-evolution.

Decreased dynamic variability of the cerebellum in the euthymic patients with bipolar disorder.

BACKGROUND: Bipolar disorder (BD) is a complex mental illness characterized by different mood states, including depression, mania/hypomania, and euthymia. This study aimed to comprehensively evaluate dynamic changes in intrinsic brain activity by using dynamic fractional amplitude of low-frequency fluctuations (dfALFF) and dynamic degree centrality (dDC) in patients with BD euthymia or depression and healthy individuals. METHODS: The resting-state functional magnetic resonance imaging data were analyzed from 37 euthymic and 28 depressed patients with BD, as well as 85 healthy individuals. Using the sliding-window method, the dfALFF and dDC were calculated for each participant. These values were compared between the 3 groups using one-way analysis of variance (ANOVA). Additional analyses were conducted using different window lengths, step width, and window type to ensure the reliability of the results. RESULTS: The euthymic group showed significantly lower dfALFF and dDC values of the left and right cerebellum posterior lobe compared with the depressed and control groups (cluster level PFWE < 0.05), while the latter two groups were comparable. Brain regions showing significant group differences in the dfALFF analysis overlapped with those with significant differences in the dDC analysis. These results were consistent across different window lengths, step width, and window type. CONCLUSIONS: These findings suggested that patients with euthymic BD exhibit less flexibility of temporal functional activities in the cerebellum posterior lobes compared to either depressed patients or healthy individuals. These results could contribute to the development of neuropathological models of BD, ultimately leading to improved diagnosis and treatment of this complex illness.

Search for serum biomarkers in patients with bipolar disorder and major depressive disorder using metabolome analysis.

Objective: Bipolar disorder (BD) and major depressive disorder (MDD) are two common psychiatric disorders. Due to the overlapping clinical symptoms and the lack of objective diagnostic biomarkers, bipolar disorder (BD) is easily misdiagnosed as major depressive disorder (MDD), which in turn affects treatment decisions and prognosis. This study aimed to investigate biomarkers that could be used to differentiate BD from MDD. Methods: Nuclear magnetic resonance (NMR) spectroscopy was performed to assess serum metabolic profiles in depressed patients with BD (n = 59), patients with MDD (n = 14), and healthy controls (n = 10). Data was analyzed using partial least squares discriminant analysis, orthogonal partial least squares discriminant analysis and t-tests. Different metabolites (VIP > 1 and p < 0.05) were identified and further analyzed using Metabo Analyst 5.0 to identify relevant metabolic pathways. Results: The metabolic phenotypes of the BD and MDD groups were significantly different from those of the healthy controls, and there were different metabolite differences between them. In the BD group, the levels of 3-hydroxybutyric acid, n-acetyl glycoprotein, ß-glucose, pantothenic acid, mannose, glycerol, and lipids were significantly higher than those in the healthy control group, and the levels of lactate and acetoacetate were significantly lower than those in the healthy control group. In the MDD group, the levels of 3-hydroxybutyric acid, n-acetyl glycoprotein, pyruvate, choline, acetoacetic acid, and lipids were significantly higher than those of healthy controls, and the levels of acetic acid and glycerol were significantly lower than those of healthy controls. Conclusion: Glycerolipid metabolism is significantly involved in BD and MDD. Pyruvate metabolism is significantly involved in MDD. Pyruvate, choline, and acetate may be potential biomarkers for MDD to distinguish from BD, and pantothenic acid may be a potential biomarker for BD to distinguish from MDD.

Identifying potential insecticide resistance markers through genomic-level comparison of Bemisia tabaci (Gennadius) lines.

The invasive whitefly (Bemisia tabaci) MED is one of the most economically damaging plant pests. The extensive use of insecticide over decades has led to that the invasive B. tabaci MED has developed resistance to a wide range of insecticide classes, but little is known about the genetic background associated with resistance. To this end, we conducted a comparative genome-wide analysis of single-base nucleotide polymorphisms between MED whitefly lines collected from fields that were recently infested and an insecticide-susceptible MED whitefly line collected in 1976. First, low-coverage genome sequencings were conducted on DNA isolated from individual whiteflies. The sequencing results were evaluated using an available B. tabaci MED genome as a reference. Significant genetic differences were discovered between MED whitefly lines collected from fields that were recently infested and an insecticide-susceptible MED whitefly line based on the principal component analyses. Top GO categories and KEGG pathways that might be involved in insecticide resistance development were identified, and several of them have not been previously associated with resistance. Additionally, we identified several genetic loci with novel variations including Cytochrome P450 monooxygenases (P450s), UDP-glucuronosyltransferases (UGTs), Glutathione S-transferases (GSTs), esterase, carboxyl-esterases (COE), ABC transporters, fatty acyl-CoA reductase, voltage-gated sodium channels, GABA receptor, and cuticle proteins (CPs) that were previously reported to have close associations with pesticide resistance in well-studied insect groups that provide an essential resource for the design of insecticide resistance-linked loci arrays insecticide. Our results was obtained solely on resequencing genome data sets, more pesticide bio-assays combined with omics datasets should be further used to verify the markers identified here.

Core of sensory gating deficits in first-episode schizophrenia: attention dysfunction.

Background: Sensory gating deficits are a common feature of schizophrenia and may be indicative of higher-order psychopathological impairments. It has been proposed that incorporating subjective attention components into prepulse inhibition (PPI) measures may improve the accuracy of assessing these deficits. This study aimed to investigate the relationship between modified PPI and cognitive function, with a specific focus on subjective attention, to gain a better understanding of the underlying mechanisms of sensory processing deficits in schizophrenia. Methods: Fifty-four unmedicated first-episode schizophrenia (UMFE) patients and 53 healthy controls participated in this study. The modified Prepulse Inhibition paradigm, including Perceived Spatial Separation PPI (PSSPPI) and Perceived Spatial Colocation PPI (PSCPPI), was used to evaluate sensorimotor gating deficits. Cognitive function was assessed in all participants using the Chinese version of the MATRICS Consensus Cognitive Suite Test (MCCB). Results: UMFE patients had lower MCCB scores and deficient PSSPPI scores than healthy controls. PSSPPI was negatively correlated with total PANSS scores and positively correlated with the speed of processing, attention/ vigilance, and social cognition. Multiple linear regression analysis showed that the PSSPPI at 60 ms had a significant effect on attentional/ vigilance and social cognition, even after controlling for gender, age, years of education, and smoking. Conclusion: The study revealed notable impairments in sensory gating and cognitive function in UMFE patients, best reflected by the PSSPPI measure. Specifically, PSSPPI at 60 ms was significantly associated with both clinical symptoms and cognitive performance, suggesting that PSSPPI at 60 ms may capture psychopathological symptoms related to psychosis.

Common and unique alterations of functional connectivity in major depressive disorder and bipolar disorder.

OBJECTIVE: Major depressive disorder (MDD) and bipolar disorder (BD) are considered whole-brain disorders with some common clinical and neurobiological features. It is important to investigate neural mechanisms to distinguish between the two disorders. However, few studies have explored the functional dysconnectivity between the two disorders from the whole brain level. METHODS: In this study, 117 patients with MDD, 65 patients with BD, and 116 healthy controls completed resting-state functional magnetic resonance imaging (R-fMRI) scans. Both edge-based network construction and large-scale network analyses were applied. RESULTS: Results found that both the BD and MDD groups showed decreased FC in the whole brain network. The shared aberrant network across patients involves the visual network (VN), sensorimotor network (SMN), dorsal attention network (DAN), and ventral attention network (VAN), which is related to the processing of external stimuli. The default mode network (DMN) and the limbic network (LN) abnormalities were only found in patients with MDD. Furthermore, results showed the highest decrease in edges of patients with MDD in between-network FC in SMN-VN, whereas in VAN-VN of patients with BD. CONCLUSIONS: Our findings indicated that both MDD and BD are extensive abnormal brain network diseases, mainly aberrant in those brain networks correlated to the processing of external stimuli, especially the attention network. Specific altered functional connectivity also was found in MDD and BD groups, respectively. These results may provide possible trait markers to distinguish the two disorders.

White matter microstructural abnormalities in individuals with attenuated positive symptom syndromes.

White matter (WM) microstructural alterations have been extensively studied in patients with psychosis, but research on the microstructure of WM in individuals with attenuated positive symptom syndrome (APSS) is currently limited. To improve the understanding of the neuropathology in APSS, this study investigated the WM of individuals with APSS using diffusion tensor and T1-weighted imaging. Automated fiber quantification was used to calculate the diffusion index values along the trajectories of 20 major fiber tracts in 42 individuals with APSS and 51 age-and sex-matched healthy control (HC) individuals. The diffusion index values in each of fiber tracts were compared node-by-node between the 2 groups. Compared with the HC group, the APSS group showed differences in the diffusion index values in partial segments of the callosum forceps minor, left and right cingulum cingulate, inferior fronto-occipital fasciculus, right corticospinal tract, left superior longitudinal fasciculus, and arcuate fasciculus. Notably, in the APSS group positive associations were found between the axial diffusivity values of the partial nodes of the left and right cingulum cingulate and the current Global Assessment of Functioning scores, as well as between the axial diffusivity values of the partial nodes of the right corticospinal tract and negative symptoms scores and reasoning and problem-solving scores. These findings suggest that individuals with APSS exhibit reduced WM integrity or possible impaired myelin in certain segments of WM tracts involved in the frontal- and limbic-cortical connections. Additionally, abnormal WM tracts appear to be associated with impaired general function and neurocognitive function. This study provides important new insights into the neurobiology of APSS and highlights potential targets for future intervention and treatment.

Efficacy of preemptive intercostal nerve block on recovery in patients undergoing video-assisted thoracic lobectomy.

BACKGROUND: Preemptive intercostal nerve block (pre-ICNB) achieves the same analgesic effects as postoperative ICNB (post-ICNB) remains unclear. This study aimed to evaluate the efficacy of preemptive ICNB on perioperative outcomes for patients undergoing video-assisted thoracic surgery (VATS). METHODS: This was a randomized, open-label study (ChiCTR2200055667) from August 1, 2021, to December 30, 2021. Eligible patients scheduled for lobectomy for lung cancer were allocated into the pre-ICNB group and the post-ICNB group. The postoperative pain evaluation, patient rehabilitation, and opioid consumption were observed. RESULTS: A total of 81 patients were included. When compared with the post-ICNB group, the pre-ICNB group had a lower proportion of hypertension comorbidity (P = 0.023), significantly lower total consumption of morphine milligram equivalents (MMEs) (P = 0.016), shorter extubation time (P = 0.019). The pre-ICNB group has similar Numeric Rating Scales (NRS) scores of dynamic pain in the post-anesthesia care unit (PACU), postoperative 6 h, 12 h, 24 h, and 48 h (P > 0.05), and had simialr scores of Bruggrmann Comfort Scale (BCS) in postoperative 6 h, 12 h, 24 and 48 h (P > 0.05). The scores of the Mini-mental state examination (MMSE) and Ramsay in the pre-ICNB group were comparable to those in the post-ICNB group, except the scores of MMSE and Ramsay in postoperative 6 h were lower (P = 0.048 and P = 0.019). The pain evaluation in the 1-month follow-up was comparable with that in the post-ICBN group (P > 0.05). CONCLUSIONS: Pre- ICNB is equally efficacious in perioperative pain management as post-ICNB, and pre-ICNB significantly reduces intra-operative opioid consumption, providing faster recovery in PACU. TRIAL REGISTRATION: Registered in the Chinese Clinical Trial Register (ChiCTR2200055667).

Altered Dynamic Functional Connectivity in Early Psychosis Between the Salience Network and Visual Network.

In early psychosis there are alterations in the static functional interaction between the salience network (SN) and higher-order cognitive networks. It is unclear whether these changes extend to the dynamic functional connectivity (dFC) of the SN, and whether the dFC between the SN and low-order networks (e.g., sensory networks) is affected. This study examined the temporal properties of the functional connectivity of the SN in individuals with early psychosis. In this study, selected core regions of the SN included the left and right anterior insula (AIs) and anterior cingulate cortex (ACC), based on independent component analysis of 26 and 20 subjects with first-episode schizophrenia (FES) or at clinical high risk for psychosis (CHR), and 37 healthy controls (HC). The dFC of each region was investigated via sliding window-based analyses with whole brain voxels. We found compared with the HC, in the FES and CHR groups the bilateral AI and ACC showed less variability in dFC with regions in the visual network; the variability between the ACC and visual regions in the FES group was less than that of the CHR; and in the FES and CHR groups the variability in dFC was higher between the right AI and the left precuneus (a core region of the default mode network). This study confirmed abnormality of dynamic functional interaction between the SN and the DMN in psychosis. More importantly, the disruption of communication between the SN and the lower-order brain network is another important aspect of the neural basis of psychosis.

Metabolomic analysis to detect urinary molecular changes associated with bipolar depression.

Bipolar disorder (BD) is a debilitating mental disorder with complex clinical manifestations and low diagnostic accuracy. Depressive episodes are most common in the course of BD with high comorbidity and suicide rates, which present greater clinical challenges than mania and hypomania episodes. However, there are no objective biomarkers for bipolar depression. The aim of this study was to detect urinary metabolite biomarkers that could be useful for the diagnosis of bipolar depression. Nuclear magnetic resonance spectroscopy was used to profile urine samples of patients with bipolar depression (n = 37) and healthy volunteers (n = 48). Data were analyzed using Orthogonal Partial Least Square Discriminant Analysis and t-test. Differential metabolites were identified (VIP > 1 and p < 0.05), and further analyzed using Metabo Analyst 3.0 to identify associated metabolic pathways. In total, we identified seven metabolites differentially expressed in patients with BD and healthy controls. Compared with healthy group, the levels of betaine, glycerol, hippuric acid, indole sulfate, trimethylamine oxide, and urea in urine samples of BD patients were significantly higher, while the level of inositol was significantly lower. Most of these small molecules are related to lipid metabolism and gut microbiota metabolism. These differential metabolites could provide critical insight into the pathological mechanisms of bipolar depression. The results of this study provide a meaningful reference for similar and further studies in the future.

Expression And Biological Interaction Network Of RHOC For Hepatic Carcinoma With Metastasis In PBMC Samples.

OBJECTIVES: Hepatic carcinoma with metastasis remains incurable, and clinical diagnostic methods lacked adequate sensitivity and specificity. Therefore, seeking effectively diagnostic biomarkers is still essential for it. RHOC was reported to be linked to metastasis of hepatic carcinoma. However, almost all of the studies used tissues as detection samples, which was not ideal for clinical course minoring. Therefore, here, it was aimed to use PBMC samples that were not only easily accessible but also minimally invasive to determine the expression and biological interaction network of RHOC for hepatic carcinoma with metastasis. METHODS: PBMC samples were isolated. Then, RNA-seq was performed to identify the DEGs between hepatic carcinoma with metastasis and hepatic carcinoma with solitary tumor. Subsequently, q-RT-PCR was used to verify the expression level of RHOC. Finally, bioinformatic analysis was used to present the biological interaction network of RHOC for hepatic carcinoma with metastasis in PBMC samples. RESULTS: The results of both RNA-seq and q-RT-PCR showed that the expression level of RHOC was significantly higher in the PBMC samples of hepatic carcinoma with metastasis than in those of hepatic carcinoma with solitary tumor. By using variety of bioinformatic analysis platforms, in PBMCs, 18 co-expression genes with RHOC were identified and their interaction network showed that MYL9 and RHOC had the highest edge evidence, and were involved in some cell migration-related pathways. CONCLUSION: Our results indicated that RHOC in PBMCs could be potentially minimally invasive indicators for the diagnosis and clinical course supervision of hepatic carcinoma with metastasis, and its biological interaction network determined based on bioinformatic methods would lay a foundation for further study of the role of RHOC in tumor invasion and metastasis.

Effect of different thoracic anesthesia on postoperative cough.

BACKGROUND: The objective of the study is to retrospectively analyze the cough status after double lumen tube (DLT) and spontaneous respiration thoracic anesthesia, to compare the degree of influence of anesthesia and surgical factors, and to investigate whether spontaneous respiration anesthesia can reduce the incidence of cough. METHODS: Postoperative follow-ups were performed on 1,162 patients from July 2011 to December 2015 who meet the selected conditions, whose surgical approach is limited to VAST bullectomy, wedge resection, segmentectomy, or lobectomy. Patients' probability of cough in 1st day (T1), 2nd days (T2), 3rd days (T3), 1st month (T4), 3rd months (T5), 6th months (T6) and 12th months (T7) after thoracoscopic surgery were recorded, as well as the Leicester cough questionnaire (LCQ) survey results, visual cough score (VAS), and cough symptom scores. All cases were divided into double-lumen endotracheal tubes anesthesia group (group T, n=925 cases) and spontaneous respiratory anesthesia group (group S, n=456 cases), and group S was further divided into intravenous composite intercostal nerve block anesthesia group (group SB, n=157 cases) and intravenous combined epidural anesthesia group (group SE, n=299 cases). RESULTS: The probability of cough decreases with the increasing of postoperative time (P<0.05). The probability of cough is similar between group SE and group SB (P>0.05). The probability of cough in group T is significantly higher than other groups at any time point (P<0.05). In group T, the symptom of cough is the most severe, the scores of physiological, psychological, and social parts of LCQ are the lowest, and the VAS score is the highest (P<0.05), but all these are similar in group SE and group SB (P>0.05). The duration of antibiotic application, the days of chest drainage tube indwelling, and the days of hospital stay are all lower in group S than in group T (P<0.05). CONCLUSIONS: There is a correlation between pulmonary surgery and postoperative cough. The probability of postoperative cough is higher in the more invasive patients. The probability of coughing is approximately 27% to 36% at 3 months after surgery, and approximately 2.6% to 7.9% in one year after surgery. The combination of surgery and anesthesia methods increases the probability of cough from 48.9% to 65.1% at 3 months after surgery, and about 20.5% to 22.8% in 1 year after surgery. Spontaneous respiration anesthesia can significantly reduce the probability of cough, improve postoperative recovery, and improve postoperative quality of life.

Impact of RNA integrity and blood sample storage conditions on the gene expression analysis.

BACKGROUND: The reliability of RNA sequencing (RNA-seq) output is affected by the quality of RNAs, which is in turn dependent on the quality of samples. Therefore, the purposes of this study were to reconsider the threshold of the RNA integrity number (RIN) and propose a simple and efficient storage scheme of blood samples for RNA-seq. PATIENTS AND METHODS: The RNAs were extracted from blood samples that were stored at different conditions and used for sequencing. The bioinformatic analyses were performed to evaluate the impact of RNA integrity and blood sample storage conditions on the gene expression analysis. RESULTS: Our outcomes showed that the samples with RIN values more than 5.3 scarcely affected the quantitative results of RNA-seq, and the influence of inherent cellular physiological processes on RNA-seq output could be negligible. CONCLUSION: The blood samples stored at 4°C within 7 days with RIN values more than 5.3 were available for RNA-seq.

Metatranscriptomics analysis of cyanobacterial aggregates during cyanobacterial bloom period in Lake Taihu, China.

Molecular mechanism of interaction between the bloom-forming cyanobacterial species and attached microbios within cyanobacterial aggregates has not been elucidated yet and understanding of which would help to unravel the cyanobacteria bloom-forming mechanism. In this study, we profiled the metabolically active community by high-throughput metatranscriptome sequencing from cyanobacterial aggregates during cyanobacterial bloom period in Lake Taihu, China. A total of 308 million sequences were obtained using the HiSeq 2500 sequencing platform, which provided a great sequence coverage to carry out the in-depth taxonomic classification, functional classification, and metabolic pathway analysis of the cyanobacterial aggregates. The results show that bacteria dominated in cyanobacterial aggregates, accounting for more than 96.66% of total sequences. Microcystis was the most abundant genus, accounted for 26.80% of total assigned sequences at the genus level in cyanobacterial aggregates community; however, Proteobacteria (46.20%) was found to be as the most abundant active bacterial populations at the phylum level. More importantly, nitrogen, phosphonate, and phosphinate metabolism which associated with eutrophication were found in this study. Especially, the enzymes and organisms relating to denitrification and anammox of nitrogen metabolism, which reduced nitrogen concentration by reducing nitrate to nitrogen to inhibit the eutrophication, were first discovered in Lake Taihu during cyanobacterial bloom period. The present study provides a snapshot of metatranscriptome for cyanobacterial aggregates in Lake Taihu and demonstrates that cyanobacterial aggregates could play a key role in the nitrogen cycle in eutrophic water.

Detection of 22 common leukemic fusion genes using a single-step multiplex qRT-PCR-based assay.

BACKGROUND: Fusion genes generated from chromosomal translocation play an important role in hematological malignancies. Detection of fusion genes currently employ use of either conventional RT-PCR methods or fluorescent in situ hybridization (FISH), where both methods involve tedious methodologies and require prior characterization of chromosomal translocation events as determined by cytogenetic analysis. In this study, we describe a real-time quantitative reverse transcription PCR (qRT-PCR)-based multi-fusion gene screening method with the capacity to detect 22 fusion genes commonly found in leukemia. This method does not require pre-characterization of gene translocation events, thereby facilitating immediate diagnosis and therapeutic management. METHODS: We performed fluorescent qRT-PCR (F-qRT-PCR) using a commercially-available multi-fusion gene detection kit on a patient cohort of 345 individuals comprising 108 cases diagnosed with acute myeloid leukemia (AML) for initial evaluation; remaining patients within the cohort were assayed for confirmatory diagnosis. Results obtained by F-qRT-PCR were compared alongside patient analysis by cytogenetic characterization. RESULTS: Gene translocations detected by F-qRT-PCR in AML cases were diagnosed in 69.4% of the patient cohort, which was comparatively similar to 68.5% as diagnosed by cytogenetic analysis, thereby demonstrating 99.1% concordance. Overall gene fusion was detected in 53.7% of the overall patient population by F-qRT-PCR, 52.9% by cytogenetic prediction in leukemia, and 9.1% in non-leukemia patients by both methods. The overall concordance rate was calculated to be 99.0%. Fusion genes were detected by F-qRT-PCR in 97.3% of patients with CML, followed by 69.4% with AML, 33.3% with acute lymphoblastic leukemia (ALL), 9.1% with myelodysplastic syndromes (MDS), and 0% with chronic lymphocytic leukemia (CLL). CONCLUSIONS: We describe the use of a F-qRT-PCR-based multi-fusion gene screening method as an efficient one-step diagnostic procedure as an effective alternative to lengthy conventional diagnostic procedures requiring both cytogenetic analysis followed by targeted quantitative reverse transcription (qRT-PCR) methods, thus allowing timely patient management.

No Significant Difference between Plasma miRNAs and Plasma-Derived Exosomal miRNAs from Healthy People.

BACKGROUND: MicroRNAs (miRNAs) are small noncoding RNAs that are involved in many biological regulation processes. Studies have reported that miRNAs are enriched in human plasma and plasma-derived exosome as novel diagnostic biomarkers. The aim of this study was to determine whether the miRNA expression levels are different between plasma and plasma-derived exosome. METHODS: We sequenced and quantified the miRNAs in plasma and exosome from healthy blood samples and validated three miRNAs in the two groups of lung cancer samples by qRT-PCR. RESULTS: The sequencing results showed that only several of miRNAs were differential, while the qRT-PCR further validated that most of them did not have the consistent differences. However, the levels of two upregulated miRNAs (miR-181b-5p and miR-21-5p) in lung cancer were significantly higher in exosomes than plasma. CONCLUSIONS: To the best of our knowledge, this study is the first to compare the expression levels of miRNAs between plasma and exosome in healthy blood samples. Our data suggested that the miRNA levels were similar in the two parts of the healthy people, whereas the two onco-miRNAs were significantly enriched in the exosome of lung cancer patients.

Construction and optimization of an efficient breathing-based isothermal emulsion amplification method.

The reaction temperature is one of the main factors that affect the stability of emulsion PCR (emPCR). Focusing on this point, we applied the "DNA breathing" mechanism in BEAMing (Bead, Emulsion, Amplification, and Magnetic) and proposed a more stable emulsion amplification method. Compared to the conventional emPCR, this method provided excellent results. Firstly, more stable emulsion system resulted in higher percentage of single-molecular amplifications (73.17%). Secondly, an ordinary temperature-controlling device was enough. Our outcome showed that the reaction temperature of this method was not strict so that the ordinary temperature-controlling device was enough for it (the heat block sets vs. the PCR instrument: 13.140 ± 0.110 vs. 13.008 ± 0.039, P = 0.120). Thirdly, the single-biotinylated emP1 coated streptavidin beads were stable enough to be used for this method (the control temperature vs. the reaction temperature: 2967.91 ± 409.045 vs. 3026.22 ± 442.129, P = 0.334), which could replace the double-biotinylated emP1 coated beads and was benefit for saving cost. In conclusion, the method presented here with stable emulsion system, simplified temperature-controlling device, and decreased investment would be a highly streamlined and inexpensive option for future single-molecular amplification based researches.

[Effects of Atorvastatin on Proliferation and Apoptosis of Leukemia Cell Line HL-60 and Its Mechanism of Signal Pathway].

OBJECTIVE: To investigate the effect of atorvastatin on proliferation and apoptosis of leukemia cell line HL-60 and its mechanism of signal pathway. METHODS: The leukemia HL-60 cells in logarithmic growth phase were seeded in 96 well plates and were treated with 1, 5 and 10 mol/L atorvastatin, then were cultured in the incubator (at 37 °C, 5% CO2) for 12 h, 24 h, 48 h. MTT colorimetric method was used to detect the proliferation leukemia cells, the apoptosis of leukemia cells was detected by flow cytometry; the expresion levels of phosphatidylinositol 3-kinase(PI3K), serine threonine protein kinase(ATK) and mTOR at mRNA and protein levels were detected by RT-PCR and Western blot respectively. The experiments included blank control group, the negative control group and drug-treated group. RESULTS: Atorvastatin could inhibit the proliferation of HL-60 cells. The treatment of HL-60 cells with 10 mol/L atorvastatin for 48 hours showed the strongest inhibition rate (39.78±3.00)% which was statistically significant different from negative control group (t=4.015, P<0.05) and the strongest induction-apoptosis effect on HL-60 cells (43.30±3.92)%, that was statistically significantly different from negative control group (t=3.624, P<0.05). After treatment with atorvastatin for 48 hours, the expression levels of PI3K,ATK and mTOR were decreased, in which the effect of 10 mol/L atorvastatin was the most obvious; The expression levels of PI3K,ATK and mTOR were decreased by (37.04±4.15)%, (53.81±3.25)% and (40.62±2.41) respectively, significantly different from the negative control (t=4.806,3.800,4.313, P<0.05). CONCLUSION: Atorvastatin may inhibit the proliferation of HL-60 cells and induce apoptosis by inhibiting the PI3K/ATK/mTOR signaling pathway.

Screening effective differential expression genes for hepatic carcinoma with metastasis in the peripheral blood mononuclear cells by RNA-seq.

Tumor metastasis is a multistep process involving a number of genetic alterations so that the genetic diagnosis is got increasingly attentions today. The aim of this study was to use RNA-seq to screen the effective differential expression genes in the peripheral blood mononuclear cells for the hepatic carcinoma with metastasis. The results showed that hepatic carcinoma samples gathered according to different metastasis. CCL3, CCL3L1, JUN, IL8, and IL1B were identified in inflammation mediated by chemokine and cytokine signaling pathway (P00031) in the hepatic carcinoma samples with metastasis, and subsequently confirmed by quantitative real-time polymerase chain reaction. In conclusions, CCL3, CCL3L1, JUN, IL8, and IL1B have the potential to be considered as candidates for future molecular diagnosis of the hepatic carcinoma with metastasis. This work may provide us with new visions into the metastasis process and potential efficient clinical diagnosis in the future.

Targeted radionuclide therapy for lung cancer with iodine-131-labeled peptide in a nude-mouse model.

Integrin α3ß1 has been shown to be a novel candidate target for the imaging and specific therapy of non-small-cell lung cancer. We have previously reported on a peptide containing a novel motif of NGXG that specifically binds to the integrin α3 receptor on lung cancer cells using a one-bead one-peptide combinatorial library. In this study, we developed the peptide cNGEGQQc-based therapeutic agent labeling with radionuclide iodine-131 (I) and evaluated its characteristics including stability, biodistribution, antitumor activity, and safety. The results showed that I-cNGEGQQc was stable in serum. Furthermore, the biodistribution of I-cNGEGQQc was determined in normal mice and rabbits. In-vivo biodistribution studies showed that radiolabeled peptide in the kidney was significantly higher than that in other organs. Nude mice bearing lung cancer cell xenografts (H1975 and L78) were used as an in-vivo model for tumor-inhibition efficacy studies with I-cNGEGQQc. The tumor growth decreased significantly in mice receiving I-labeled peptide compared with the controls and the effect of I-labeled peptide can be blocked by unlabeled cNGEGQQc. Safety studies showed that I-cNGEGQQc was relatively safe for animals without significant toxicity. Our data suggest that I-cNGEGQQc has potential as a targeted radiotherapeutic agent for non-small-cell lung cancer.