Complete Genome Sequence Analysis of Bacillus subtilis MC4-2 Strain That against Tobacco Black Shank Disease.

The MC4-2 bacterium strain was isolated and purified from the Periplaneta americana intestine as a biocontrol agent with good antagonistic effect against the pathogens of a soil-borne disease called tobacco black shank. The MC4-2 strain was found to have good broad-spectrum inhibition by plate stand-off test. Based on 16S rRNA and gyrB genes, ANI analysis, and other comparative genomics methods, it was determined that the MC4-2 strain was Bacillus subtilis. The complete genome sequence showed that the genome size was 4,076,630 bp, the average GC content was 43.78%, and the total number of CDSs was 4,207. Genomic prediction analysis revealed that a total of 145 genes were annotated by the CAZy, containing mainly GH and CE enzymes that break down carbohydrates such as glucose, chitin, starch, and alginate, and a large number of enzymes involved in glycosylation were present. A total of ten secondary metabolite clusters were predicted, six clusters of which were annotated as surfactin, bacillaene, fengycin, bacillibactin, subtilosin A, and bacilysin. The present investigation found the biological control mechanism of B. subtilis MC4-2, which provides a strong theoretical basis for the best use of this strain in biological control methods and provides a reference for the subsequent development of agents of this bacterium.

Comparative proteomic analysis reveals insights into the response of Cryptolaemus montrouzieri to bottom-up transfer of cadmium and lead across a multi-trophic food chain.

Contamination of agro-ecosystems with heavy metals can affect the development and reproduction of insect natural enemies. This study reports a detailed Tandem Mass Tag based quantitative proteomic analysis of underlying mechanisms responsible for stress response of Cryptolaemus montrouzieri against heavy metals (cadmium (Cd) and lead (Pb)) transported across a multi-trophic food chain. A total of 6639 proteins were detected under Cd as well as Pb stress. In Pb versus the control cluster, 69 proteins (28 up-regulated and 41 down-regulated) were differentially expressed whereas 268 proteins were differentially expressed under Cd versus the control cluster, having 198 proteins up-regulated and 70 down-regulated proteins. The analysis of differentially expressed proteins showed that 27 proteins overlapped in both clusters representing the core proteome to Pb and Cd stress. The bioinformatics analysis demonstrated that these proteins were mapped to 57 and 99 pathways in Pb versus control and Cd versus control clusters, respectively. The functional classification by COG, GO and KEGG databases showed significant changes in protein expression by C. montrouzieri under Pb and Cd stress. The heavy metal stress (Pb and Cd) induced significant changes in expression of proteins like hexokinase (HK), succinyl-CoA, trypsin like proteins, cysteine proteases, cell division cycle proteins, and yellow gene proteins. The results provide detailed information on the protein expression levels of C. montrouzieri and will serve as basic information for future proteomic studies on heavy metal responses of insect predators within a multi-trophic food chain.

Meta-Analysis of Pulmonary Artery Denervation for Treatment of Pulmonary Hypertension.

INTRODUCTION: Pulmonary artery denervation (PADN) can reduce the sympathetic nervous system (SNS) activity, reduce pulmonary artery pressure (PAP), and improve the quality of life in patients with pulmonary hypertension (PH). We conducted a systematic meta-analysis of the effectiveness of PADN in the treatment of PH patients. METHODS: This is a comprehensive literature search including all public clinical trials investigating the effects of PADN on PH. Outcomes were mean pulmonary artery pressure (mPAP), pulmonary vascular resistance (PVR), cardiac output (CO), right ventricular (RV) Tei index, 6-minute walk distance (6MWD), and New York Heart Association (NYHA) cardiac function grading. RESULTS: A total of eight clinical studies with 213 PH patients who underwent PADN were included. Meta-analysis showed that after PADN, mPAP (mean difference [] -12.51, 95% confidence interval [CI] -17.74 to -7.27, P<0.00001) (mmHg) and PVR ( -5.17, 95% CI -7.70 to -2.65, P<0.0001) (Wood unit) decreased significantly, CO ( 0.59, 95% CI 0.32 to 0.86, P<0.0001) (L/min) and 6MWD ( 107.75, 95% CI 65.64 to 149.86, P<0.00001) (meter) increased significantly, and RV Tei index ( -0.05, 95% CI -0.28 to 0.17, P=0.63) did not change significantly. Also after PADN, the proportion of NYHA cardiac function grading (risk ratio 0.23, 95% CI 0.14 to 0.37, P<0.00001) III and IV decreased significantly. CONCLUSION: This meta-analysis supports PADN as a potential new treatment for PH. Further high-quality randomized controlled studies are needed.

Meta-Analysis of Pulmonary Artery Denervation for Treatment of Pulmonary Hypertension

ABSTRACT Introduction: Pulmonary artery denervation (PADN) can reduce the sympathetic nervous system (SNS) activity, reduce pulmonary artery pressure (PAP), and improve the quality of life in patients with pulmonary hypertension (PH). We conducted a systematic meta-analysis of the effectiveness of PADN in the treatment of PH patients. Methods: This is a comprehensive literature search including all public clinical trials investigating the effects of PADN on PH. Outcomes were mean pulmonary artery pressure (mPAP), pulmonary vascular resistance (PVR), cardiac output (CO), right ventricular (RV) Tei index, 6-minute walk distance (6MWD), and New York Heart Association (NYHA) cardiac function grading. Results: A total of eight clinical studies with 213 PH patients who underwent PADN were included. Meta-analysis showed that after PADN, mPAP (mean difference [MD] -12.51, 95% confidence interval [CI] -17.74 to -7.27, P<0.00001) (mmHg) and PVR (MD -5.17, 95% CI -7.70 to -2.65, P<0.0001) (Wood unit) decreased significantly, CO (MD 0.59, 95% CI 0.32 to 0.86, P<0.0001) (L/min) and 6MWD (MD 107.75, 95% CI 65.64 to 149.86, P<0.00001) (meter) increased significantly, and RV Tei index (MD -0.05, 95% CI -0.28 to 0.17, P=0.63) did not change significantly. Also after PADN, the proportion of NYHA cardiac function grading (risk ratio 0.23, 95% CI 0.14 to 0.37, P<0.00001) III and IV decreased significantly. Conclusion: This meta-analysis supports PADN as a potential new treatment for PH. Further high-quality randomized controlled studies are needed.

Clinical significance of LAG-3 on microvessel density in primary hepatocellular carcinoma.

Aims: We aimed to determine whether lymphocyte activation gene 3 (LAG-3), also known as CD223, is associated with microvessel density (MVD) in primary hepatocellular carcinoma (HCC), as well as their clinical significance in predicting survival. Materials and methods: One hundred and twenty-seven patients were enrolled in the study. Samples were obtained on resection at the Department of Hepatobiliary Surgery of the Qingdao Municipal Hospital from June 2014 to June 2016. Immunohistochemistry was used to determine vessel density and LAG-3 abundance. Statistical analyses were performed to test for correlation of LAG-3 density and other clinicopathological variables with overall survival (OS). Results: High LAG-3 abundance was significantly correlated with increased MVD in primary HCC (P < 0.05). The χ2 test revealed a significant association of LAG-3 with preoperative AFP level, tumor diameter, N stage, and the presence of HBV infection (P < 0.05). Patients with high LAG-3 expression had shorter OS compared to those with low LAG-3 expression (P < 0.05). The Cox proportional hazards model showed that both higher LAG-3 and MVD density, age, the number of tumors, preoperative AFP level, tissue differentiation, Child-Pugh grade, and lymph node metastasis correlated with survival. Conclusions: High expression of LAG-3 is associated with angiogenesis and poor prognosis in HCC patients. With the deepening of research, LAG-3 is likely to become a novel biomarker for clinical diagnosis and prognosis and can even be a therapeutic target of HCC.

ß-aminobutyric acid (BABA)-induced resistance to tobacco black shank in tobacco (Nicotiana tabacum L.).

Tobacco black shank is a kind of soil-borne disease caused by the Oomycete Phytophthora parasitica. This disease is one of the most destructive diseases to tobacco (Nicotiana tabacum L.) growth worldwide. At present, various measures have been taken to control this disease, but they still have different challenges and limitations. Studies have shown that ß-aminobutyric acid (BABA), a nonprotein amino acid, can enhance disease resistance in plants against different varieties of pathogens. However, it is unclear whether BABA can induce plants to resist Phytophthora parasitica infection. Therefore, this study aims to explore the effect and related mechanism of BABA against tobacco black shank. Our results showed that 5 mmol.L-1 BABA had an obvious anti-inducing effect on the pathogenic fungus and could effectively inhibit the formation of dark spots in the stems. The results also showed that a large amount of callose deposition was observed in BABA-treated tobacco. Furthermore, the application of BABA induced the accumulation of H2O2 in tobacco and effectively regulated the homeostasis of reactive oxygen in tobacco plants, reducing the toxicity of H2O2 to plants while activating the defense system. In addition, BABA spray treatment could induce an increase in the concentrations of salicylic acid (SA) and jasmonic acid-isoleucine (JA-Ile) in tobacco, and the gene expression results confirmed that BABA upregulated the expression of SA-related genes (PR1, PR2 and PR5), JA-related genes (PDF1.2) and ET-related genes (EFE26 and ACC oxidase) in tobacco plants. Taken together, BABA could activate tobacco resistance to black shank disease by increasing H2O2 accumulation, callose deposition, plant hormone (SA and JA-Ile) production, and SA-, JA-, and ET- signaling pathways.

Integrated analysis of m6A mRNA methylation in rats with monocrotaline-induced pulmonary arterial hypertension.

BACKGROUND: N6-methyladenosine (m6A) modification is one of the most common chemical modifications of eukaryotic mRNAs, which play an important role in tumors and cardiovascular disease through regulating mRNA stability, splicing and translation. However, the changes of m6A mRNA and m6A-related enzymes in pulmonary arterial hypertension (PAH) remain largely unexplored. METHODS: MeRIP-seq was used to identify m6A methylation in lung tissues from control and MCT-PAH rats. Western blot and immunofluorescence were used to evaluate expression of m6A-related enzymes. RESULTS: Compared with control group, m6A methylation was mainly increased in lung tissues from MCT-PAH rats. The up-methylated coding genes in MCT-PAH rats were primarily enriched in processes associated with inflammation, glycolysis, ECM-receptor interaction and PDGF signal pathway, while genes with down-methylation were enriched in processes associated with TGF-ß family receptor members. The expression of FTO and ALKBH5 downregulated, METTL3 and YTHDF1 increased and other methylation modification-related proteins was not significantly changed in MCT-PAH rats lung tissues. Immunofluorescence indicated that expression of FTO decreased and YTHDF1 increased in small pulmonary arteries of MCT-PAH rats. CONCLUSION: m6A levels and the expression of methylation-related enzymes were altered in PAH rats, in which FTO and YTHDF1 may play a crucial role in m6A modification.

A Fifteen-Gene Classifier to Predict Neoadjuvant Chemotherapy Responses in Patients with Stage IB to IIB Squamous Cervical Cancer.

Neoadjuvant chemotherapy (NACT) remains an attractive alternative for controlling locally advanced cervical cancer. However, approximately 15-34% of women do not respond to induction therapy. To develop a risk stratification tool, 56 patients with stage IB-IIB cervical cancer are included in 2 research centers from the discovery cohort. Patient-specific somatic mutations led to NACT non-responsiveness are identified by whole-exome sequencing. Next, CRISPR/Cas9-based library screenings are performed based on these genes to confirm their biological contribution to drug resistance. A 15-gene classifier is developed by generalized linear regression analysis combined with the logistic regression model. In an independent validation cohort of 102 patients, the classifier showed good predictive ability with an area under the curve of 0.80 (95% confidence interval (CI), 0.69-0.91). Furthermore, the 15-gene classifier is significantly associated with patient responsiveness to NACT in both univariate (odds ratio, 10.8; 95% CI, 3.55-32.86; p = 2.8 × 10-5) and multivariate analysis (odds ratio, 17.34; 95% CI, 4.04-74.40; p = 1.23 × 10-4) in the validation set. In conclusion, the 15-gene classifier can accurately predict the clinical response to NACT before treatment, representing a promising approach for guiding the selection of appropriate treatment strategies for locally advanced cervical cancer.

Functional titanium dioxide nanoparticle conjugated with phthalocyanine and folic acid as a promising photosensitizer for targeted photodynamic therapy in vitro and in vivo.

Photodynamic therapy (PDT) is a promising cancer treatment that can be implemented using various agents. The conventional photosensitizer Al (III) phthalocyanine chloride tetrasulfonic acid (Pc) has limitations of selectivity in tumor targeting, low affinity to cancer cells, and low two-photon absorption. This study presents a novel photosensitizer FA-TiO2-Pc, which has the TiO2 nanoparticle conjugated with a tumor targeting agent of folic acid (FA), and Pc. FA-TiO2-Pc possessed high targeted photodynamic therapeutic activity and excellent biocompatibility. This promising photosensitizer showed high therapeutic drug efficiency in vitro at a low concentration dose and short incubation time under one-photon excitation (OPE). In vivo, when treated with a low dose of FA-TiO2-Pc and low light irradiation, the tumor growth was depressed in mice bearing HeLa xenograft tumors with minimal side effects. In addition, the two-photon absorption of FA-TiO2-Pc was also enhanced compared to Pc, proving that FA-TiO2-Pc system has a great potential to be used for the therapy of the folate receptor positive cancer cells in both OPE-PDT and two-photon excitation (TPE)-PDT agents.

Identification and preliminary characterization of chemosensory-related proteins in the gall fly, Procecidochares utilis by transcriptomic analysis.

Chemoreception is critical for insect behaviors such as foraging, host searching and oviposition. The process of chemoreception is mediated by a series of proteins, including odorant-binding proteins (OBPs), gustatory receptors (GRs), odorant receptors (ORs), ionotropic receptors (IRs), chemosensory proteins (CSPs) and sensory neuron membrane proteins (SNMPs). The tephritid stem gall fly, Procecidochares utilis Stone, is a type of egg parasitic insect, which is an effective biological control agent for the invasive weed Ageratina adenophora in many countries. However, the study of molecular components related to the olfactory system of P. utilis has not been investigated. Here, we conducted the developmental transcriptome (egg, first-third instar larva, pupa, female and male adult) of P. utilis using next-generation sequencing technology and identified a total of 133 chemosensory genes, including 40 OBPs, 29 GRs, 24 ORs, 28 IRs, 6 CSPs, and 6 SNMPs. The sequences of these candidate chemosensory genes were confirmed by BLAST, and phylogenetic analysis was performed. Quantitative real-time PCR (qRT-PCR) confirmed that the expression levels of the candidate OBPs varied at the different developmental stages of P. utilis with most OBPs expressed mainly in the pupae, female and male adults but scarcely in eggs and larvae, which was consistent with the differentially expressed genes (DEGs) analysis using the fragments per kilobase per million fragments (FPKM) value. Our results provide a significant contribution towards the knowledge of the set of chemosensory proteins and help advance the use of P. utilis as biological control agents.

Influence of Temperature Effect on the Static and Dynamic Performance of Gas-Lubricated Microbearings.

Gas-lubricated microbearings are widely applied in multiple fields due to their advantages of high-speed, low friction level and other features. The operating environment of microbearings is complex, and the difference of temperature has an important influence on their comprehensive performance. In this investigation, FEM (finite element method) is employed to investigate the static, dynamic and limit characteristics of microbearings lubricated by different kinds of gas at different temperatures. The results show that the rise of temperature leads to the decline of equivalent viscosity of gas, which weakens the load capacity of microbearings, and furthermore, affects the operating stability of microbearings. The dynamic performances of microbearings at different temperatures are very different, and the two dynamic limit characteristics are more sensitive to temperature when it changes.

Label-free imaging and spectroscopy for early detection of cervical cancer.

The label-free imaging and spectroscopy method was studied on cervical unstained tissue sections obtained from 36 patients. The native fluorescence spectra of tissues are analyzed by the optical redox ratio (ORR), which is defined as fluorescence intensity ratio between NADH and FAD, and indicates the metabolism change with the cancer development. The ORRs of normal tissues are consistently higher than those of precancer or cancerous tissues. A criterion line of ORR at 5.0 can be used to discriminate cervical precancer/cancer from normal tissues. The sensitivity and specificity of the native fluorescence spectroscopy method for cervical cancer diagnosis are determined as 100% and 91%. Moreover, the native fluorescence spectroscopy study is much more sensitive on the healthy region of cervical precancer/cancer patients compared with the traditional clinical staining method. The results suggest label-free imaging and spectroscopy is a fast, highly sensitive and specific method on the detection of cervical cancer.

A case of intravascular large B cell lymphoma presenting as nodular goiter.

BACKGROUND: Intravascular large B-cell lymphoma (IVLBCL) is a subtype of diffuse large B-cell lymphoma (DLBCL) that is rare and highly aggressive and that may progressively involve many organs. CNS (central nervous system), BM (bone marrow) and skin are the most common systems involved. To date, only 2 cases of IVLBCL involving the thyroid have been reported. CASE PRESENTATION: Here, we report a case of IVLBCL involving the thyroid and accompanied by bilateral nodular goiter. In this case, a thyroid mass was identified in a physical examination of a 68-year-old male who initially presented with dyspnea accompanied by intermittent headache for approximately 1 month. Computed tomography scans revealed that the left lobar thyroid was occupied by a large, slightly lower density mass (5.8 × 4.7 × 8.4 cm). However, the patient had no hyperthyroidism or hoarseness. Levels of thyroid hormones and anti-thyroid autoantibodies in the serum were normal preoperatively. Thyroid mass resection was performed to establish a diagnosis and to relieve symptoms. CONCLUSIONS: Pathological results of the surgical specimen revealed that large atypical lymphoma cells filled the capillaries in the lesion area. Immunohistochemical staining revealed that the large-sized tumor cells were positive for CD20, PAX-5, MUM-1 and BCL-2, and were negative for CD3, CD5, CD43, CD10, CD23, CyclinD1, CD138, CD30, ALK, CD56, MPO, S-100, TTF-1, TG (thyroglobulin) and CT (calcitonin). The Ki-67 index was estimated to be approximately 85%. The patient was subsequently diagnosed as "Classical" IVLBCL non-germinal center B-cell type. The patient declined chemotherapy and died in the fifth month after operation.

Thiol Specific and Tracelessly Removable Bioconjugation via Michael Addition to 5-Methylene Pyrrolones.

5-Methylene pyrrolones (5MPs) are highly thiol-specific and tracelessly removable bioconjugation tools. 5MPs are readily prepared from primary amines in one step. 5MPs exhibit significantly improved stability under physiologically relevant conditions and cysteine specificity compared to commonly used analogues, maleimides. Michael addition of thiol to 5MPs occurs rapidly, cleanly, and does not generate a stereocenter. The conjugates efficiently release thiols via retro-Michael reaction in alkaline buffer (pH 9.5) or via thiol exchange at pH 7.5. This unique property makes 5MPs valuable for the controlled release of conjugated cargo and temporary thiol protection. The utilization of 5MPs for protein immobilization and pull-down of active complexes is illustrated using E. coli. acetohydroxyacid synthase isozyme I.

Self-assembled controllable virus-like nanorods as templates for construction of one-dimensional organic-inorganic nanocomposites.

Virus-like assemblies with controllable size and surface groups can be used as efficient templates for the controllable synthesis of CdS nanorods, which represents a new strategy for controllable preparation of one-dimensional (1D) organic-inorganic nanocomposites.

The relationship between osteoprotegerin gene polymorphisms and bone mineral density in Chinese postmenopausal women.

Previous evidence supports that the osteoprotegerin (OPG) gene is one of the most important candidate genes for influencing the pathogenesis of osteoporosis. The objective of this study was to investigate the relationship between OPG gene polymorphisms and osteoporosis in Chinese postmenopausal women. A total of 764 subjects were included in this study. The bone mineral density (BMD) in the lumbar spine (L2-4), neck hip and total hip was determined by dual-energy X-ray absorptiometry (DEXA). The g.19190C>A and g.25602A>G SNPs were detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), created restriction site PCR (CRS-PCR) and DNA sequencing methods. As for g.19190C>A, our data suggested that the BMD value of lumbar spine (L2-4), neck hip and total hip for subjects with CC genotype was significantly higher than that of CA and AA genotypes (P<0.05). No significant difference was detected between the association of g.25602A>G genotypes with spine BMD and neck hip BMD, while total hip BMD almost reached the significant level (P=0.063). These findings provide more evidence that the SNPs in OPG gene could affect BMD and osteoporosis, and the allele-A of g.19190C>A and allele-G of g.25602A>G genetic variants are associated with increased risk for osteoporosis in Chinese postmenopausal women.