SDF-1 promotes metastasis of NSCLC by enhancing chemoattraction of megakaryocytes through the PI3K/Akt signaling pathway.

Lung cancer (LC) is the leading cause of cancer-associated deaths worldwide, among which non-small-cell lung cancer (NSCLC) accounts for 80%. Stromal cell-derived factor-1 (SDF-1) inhibition results in a significant depletion of NSCLC metastasis. Additionally, SDF-1 is the only natural chemokine known to bind and activate the receptor CXCR4. Thus, we attempted to clarify the molecular mechanism of SDF-1 underlying NSCLC progression. Transwell migration, adhesion, and G-LISA assays were used to assess megakaryocytic chemotaxis in vitro and in vivo in terms of megakaryocytic migration, adherence, and RhoA activation, respectively. Western blotting was used to assess PI3K/Akt-associated protein abundances in MEG-01 cells and primary megakaryocytes under the indicated treatment. A hematology analyzer and flow cytometry were used to assess platelet counts in peripheral blood and newly formed platelet counts in Lewis LC mice under different treatments. Immunochemistry and flow cytometry were used to measure CD41+ megakaryocyte numbers in Lewis LC mouse tissue under different treatments. ELISA was used to measure serum TPO levels, and H&E staining was used to detect NSCLC metastasis.SDF-1 receptor knockdown suppressed megakaryocytic chemotaxis in Lewis LC mice. SDF-1 receptor inhibition suppressed megakaryocytic chemotaxis via the PI3K/Akt pathway. SDF-1 receptor knockdown suppressed CD41+ megakaryocyte numbers in vivo through PI3K/Akt signaling. SDF-1 receptor inhibition suppressed CD41+ megakaryocytes to hinder NSCLC metastasis. SDF-1 facilitates NSCLC metastasis by enhancing the chemoattraction of megakaryocytes via the PI3K/Akt signaling pathway, which may provide a potential new direction for seeking therapeutic plans for NSCLC.

Characteristics and prognosis of EGFR mutations in small cell lung cancer patients in the NGS era.

PURPOSE: Targeted therapy has not been effective for small cell lung cancer (SCLC) patients. Although some studies have reported on EGFR mutations in SCLC, a systematic investigation into the clinical, immunohistochemical, and molecular characteristics and prognosis of EGFR-mutated SCLCs is lacking. METHODS: Fifty-seven SCLC patients underwent next-generation sequencing technology, with 11 in having EGFR mutations (group A) and 46 without (group B). Immunohistochemistry markers were assessed, and the clinical features and first-line treatment outcomes of both groups were analyzed. RESULTS: Group A consisted primarily of non-smokers (63.6%), females (54.5%), and peripheral-type tumors (54.5%), while group B mainly comprised heavy smokers (71.7%), males (84.8%), and central-type tumors (67.4%). Both groups showed similar immunohistochemistry results and had RB1 and TP53 mutations. When treated with tyrosine kinase inhibitors (TKIs) plus chemotherapy, group A had a higher treatment response rate with overall response and disease control rates of 80% and 100%, respectively, compared to 57.1% and 100% in group B. Group A also had a significantly longer median progression-free survival (8.20 months, 95% CI 6.91-9.49 months) than group B (2.97 months, 95% CI 2.79-3.15), with a significant difference (P = 0.043). Additionally, the median overall survival was significantly longer in group A (16.70 months, 95% CI 1.20-32.21) than in group B (7.37 months, 95% CI 3.85-10.89) (P = 0.016). CONCLUSION: EGFR-mutated SCLCs occurred more frequently in non-smoking females and were linked to prolonged survival, implying a positive prognostic impact. These SCLCs shared immunohistochemical similarities with conventional SCLCs, and both types had prevalent RB1 and TP53 mutations.

Endothelial extracellular vesicles induce acute lung injury via follistatin-like protein 1.

Cardiopulmonary bypass has been speculated to elicit systemic inflammation to initiate acute lung injury (ALI), including acute respiratory distress syndrome (ARDS), in patients after cardiac surgery. We previously found that post-operative patients showed an increase in endothelial cell-derived extracellular vesicles (eEVs) with components of coagulation and acute inflammatory responses. However, the mechanism underlying the onset of ALI owing to the release of eEVs after cardiopulmonary bypass, remains unclear. Plasma plasminogen-activated inhibitor-1 (PAI-1) and eEV levels were measured in patients with cardiopulmonary bypass. Endothelial cells and mice (C57BL/6, Toll-like receptor 4 knockout (TLR4-/-) and inducible nitric oxide synthase knockout (iNOS-/-)) were challenged with eEVs isolated from PAI-1-stimulated endothelial cells. Plasma PAI-1 and eEVs were remarkably enhanced after cardiopulmonary bypass. Plasma PAI-1 elevation was positively correlated with the increase in eEVs. The increase in plasma PAI-1 and eEV levels was associated with post-operative ARDS. The eEVs derived from PAI-1-stimulated endothelial cells could recognize TLR4 to stimulate a downstream signaling cascade identified as the Janus kinase 2/3 (JAK2/3)-signal transducer and activator of transcription 3 (STAT3)-interferon regulatory factor 1 (IRF-1) pathway, along with iNOS induction, and cytokine/chemokine production in vascular endothelial cells and C57BL/6 mice, ultimately contributing to ALI. ALI could be attenuated by JAK2/3 or STAT3 inhibitors (AG490 or S3I-201, respectively), and was relieved in TLR4-/- and iNOS-/- mice. eEVs activate the TLR4/JAK3/STAT3/IRF-1 signaling pathway to induce ALI/ARDS by delivering follistatin-like protein 1 (FSTL1), and FSTL1 knockdown in eEVs alleviates eEV-induced ALI/ARDS. Our data thus demonstrate that cardiopulmonary bypass may increase plasma PAI-1 levels to induce FSTL1-enriched eEVs, which target the TLR4-mediated JAK2/3/STAT3/IRF-1 signaling cascade and form a positive feedback loop, leading to ALI/ARDS after cardiac surgery. Our findings provide new insight into the molecular mechanisms and therapeutic targets for ALI/ARDS after cardiac surgery.

Combined small cell lung cancer: current progress and unmet needs.

Combined small cell lung cancer (CSCLC) is a specific subtype of lung cancer characterized by a pathological mixture of small cell lung cancer and any subtype of non-small cell lung cancer components. Currently, our understanding of the clinicopathological features, origin, molecular characterization, treatment, and prognosis of CSCLC remains limited. CSCLCs represent examples of intratumor heterogeneity and pose challenges for accurate diagnosis. Are there any distinct clinicopathologic and molecular differences between pure SCLC and CSCLC? Furthermore, the prognostic outcomes and optimal treatments for CSCLC are urgently needed. This article aims to summarize the current biological features and clinical management of CSCLC, providing a reference for further understanding of this heterogeneous form of small cell lung cancer.

Characterization of Physicochemical Properties of Oil-in-Water Emulsions Stabilized by Tremella fuciformis Polysaccharides.

In this paper, emulsions stabilized by Tremella fuciformis polysaccharides (TFP) were prepared and the physiochemical properties were assessed. Results showed that the TFP emulsions illustrated the highest emulsifying activity (EAI) and emulsifying stability (ESI) when the concentration of TFP and oil were 0.8% and 10% (wt%). The higher pH value was in favor of the emulsifying properties, while the addition of NaCl impaired the stability, and the greater the concentration, the lower the EAI and ESI. Besides, the emulsifying and rheological properties and stability analysis were evaluated in comparison with gum arabic, pectin, and carboxymethyl cellulose emulsions. It was discovered that TFP illustrated better storage and freeze-thaw stability, which was proved by the result of zeta-potential and particle size. The rheological measurement revealed that all the emulsions behaved as pseudoplastic fluids, while TFP displayed a higher viscosity. Meanwhile, TFP emulsions tended to form a more stable network structure according to the analysis of the parameters obtained from the Herschel-Bulkley model. FTIR spectra suggested that the O-H bond could be destructed without the formation of new covalent bonds during the emulsion preparation. Therefore, this study would be of great importance for the research of emulsions stabilized by TFP as a natural food emulsifier.

Chemical constituents and pharmacological activities of medicinal plants from Rosa genus.

The genus Rosa (Rosaceae family) includes about 200 species spread in the world, and this genus shows unique advantages in medicine and food. To date, several scholars concentrated on compounds belonging to flavonoids, triterpenes, tannins, polysaccharide, phenolic acids, fatty acids, organic acids, carotenoids, and vitamins. Pharmacological effects such as antineoplastic and anti-cancer properties, anti-inflammatory, antioxidant, liver protection, regulate blood sugar, antimicrobial activity, antiviral activity, as well as nervous system protection and cardiovascular protection were wildly reported. This article reviews the chemical constituents, pharmacological effects, applications and safety evaluations of Rosa plants, which provides a reference for the comprehensive utilization of medicine and food resources and gives a scientific basis for the development of medicinal plants of the genus Rosa.

Identification and validation of a ferroptosis-related prognostic risk-scoring model and key genes in small cell lung cancer.

Background: Small cell lung cancer (SCLC) is an aggressive lung malignancy with high relapse rates and poor survival outcomes. Ferroptosis is a recently identified type of cell death caused by excessive intracellular iron accumulation and lipid peroxidation, which may mediate tumor-infiltrating immune cells to influence anti-cancer immunity. But prognostic value of ferroptosis-related genes and its relationship with the treatment response of immunotherapies in SCLC have not been elucidated. Methods: The RNA-sequencing and clinical data of SCLC patients were downloaded from the cBioPortal database. A ferroptosis-related prognostic risk-scoring model was constructed based on univariable and multivariable Cox-regression analysis. Kaplan-Meier (K-M) survival curves and receiver operating characteristics (ROC) curves were constructed to assess the sensitivity and specificity of the risk-scoring model. And the correlations between ferroptosis-related prognostic genes and immune microenvironment were explored. The IC50 values of anti-cancer drugs were downloaded from the Genomics of Drug Sensitivity in Cancer (GDSC) database and the correlation analysis with the key gene thioredoxin-interacting protein (TXNIP) was performed. In addition, immunohistochemistry (IHC) staining was employed to detect the expression of TXNIP in 20 SCLC patients who received first-line chemo-immunotherapy. Immunotherapeutic response according to iRECIST (Response Evaluation Criteria in Solid Tumours for immunotherapy trials) were recorded. Results: We constructed a risk-score successfully dividing patients in the low- and high-risk groups (with better and worse prognosis, respectively). The area under the curve (AUC) of this risk-scoring model was 0.812, showing it had good utility in predicting the prognosis of SCLC. Moreover, ferroptosis-related genes were associated with the degree of immune infiltration of SCLC. Most importantly, we found that the TXNIP expression was highly correlated with the degree of immune invasion and the efficacy of chemotherapy in combination with immunotherapy in SCLC patients. Conclusions: The ferroptosis-related prognostic risk-scoring model proposed in this study can potentially predict the prognosis of SCLC patients. TXNIP may serve as a potential biomarker to predict the prognosis and efficacy of chemotherapy combined with immunotherapy in SCLC patients.

Ulnar shortening osteotomy vs. wafer resection for ulnar impaction syndrome: A systematic review and meta-analysis.

BACKGROUND: Ulnar shortening osteotomy (USO) is the most common operation to treat ulnar impaction syndrome (UIS). An alternative to USO is the arthroscopic wafer procedure (AWP). Few studies have directly compared USO and AWP in patients with UIS. This study compared ulnar shortening osteotomy (USO) versus arthroscopic wafer procedure (AWP) for UIS treatment. MATERIALS AND METHODS: PubMed, EMBASE, the Cochrane Library, Wanfang, and CNKI were systematically searched for reports published before March 2021. The outcomes included Modified Mayo Wrist Score, Darrow's Criteria, Disability of Arm, Shoulder, and Hand (DASH), grip strength, visual analog scale (VAS) score, and time to resume manual occupation. RESULTS: Seven studies (133 and 118 patients with USO and AWP, respectively) were included. There were no differences in combined Darrow's Criteria or Modified Mayo Wrist Score, Modified Mayo Wrist Score, Darrow's Criteria, revision rate, DASH score, VAS score, and time to resume manual occupation. Grip strength was better with AWP (SMD = -0.73, 95%CI: -1.36, -0.11, P = 0.022). Differences were seen for ulna positive variation, favoring USO (WMD = -2.75, 95%CI: -5.17, -0.33, P = 0.026). CONCLUSIONS: In the surgical treatment of UIS, AWP might be associated with improved grip strength, while USO seems to show better results in treating pronounced ulna positive variation. Only evidence of moderate quality could be included in this meta-analysis.

hsa_circ_0000523/miR-let-7b/METTL3 axis regulates proliferation, apoptosis and metastasis in the HCT116 human colorectal cancer cell line.

Circular RNAs (circRNAs/circs) have gained attention as a class of potential biomarkers for the early detection of multiple cancers. However, the functions and mechanisms of circRNAs in the oncogenesis of human colorectal cancer (CRC) remain to be elucidated. The present study aimed to investigate the roles of hsa_circ_0000523 and its parental gene methyltransferase-like 3 (METTL3) in regulating cell proliferation, apoptosis and invasion in the HCT116 human CRC cell line. To uncover the regulated function of hsa_circ_0000523 in HCT116 cells, a dual-luciferase reporter assay, flow cytometry, reverse transcription-quantitative PCR, Cell Counting Kit-8 assay, cell invasion and western blot assay were used. In HCT116 cells, hsa_circ_0000523 indirectly regulated METTL3 expression by suppressing the transcription of microRNA (miR)-let-7b. The expression of METTL3 promoted cell proliferation and suppressed apoptosis. In the present study, it was found that miR-let-7b promoted cell viability and inhibited apoptosis and invasion, while circ_0000523 exerted the opposite effects. Higher levels of METTL3 expression were associated with more aggressive tumor invasion. The present results suggest that circRNAs and METTL3 may be applied for highly sensitive diagnosis of CRC and for predicting prognosis in patients who have undergone therapy.

LncRNA IGBP1-AS1 targets miR-150-5p to increase ZEB1 expression in nasopharyngeal carcinoma.

Background: Nasopharyngeal carcinoma (NPC), which is a form of cancer arising from the epithelium of the nasopharynx, remains highly prevalent, particularly in Southeast Asia and Southern China. Dysregulated long non-coding RNAs (lncRNAs) are involved in several malignancies, including the growth and aggression of tumors. LncRNA IGBP1-AS1 plays an important role in the advancement of breast cancer, but its role in NPC has not yet been explored. Methods: LncRNA IGBP1-AS1 levels in human NPC samples compared with their matched adjacent normal tissues by quantitative reverse transcription polymerase chain reaction (qRT-PCR). The proliferation alteration of NPC cells was tested in vitro. Luciferase reporter assay and RNA immunoprecipitation (RIP) were carried out to reveal the interaction between lncRNA IGBP1-AS1, miR-150-5p, and zinc finger e-box binding homeobox 1 (ZEB1). Results: In the study, we found that IGBP1-AS1 was found to be significantly upregulated in the NPC samples and cell lines. The knockdown of IGBP1-AS1 impeded in-vitro proliferation of the NPC cells. Bioinformatics and reporter assays revealed an association between IGBP1-AS1 and miR-150-5p. The expression of ZEB1 was increased by the microRNA (miRNA) sequestering in human NPC. Conclusions: The progression of NPC tumors is facilitated by lncRNA IGBP1-AS1 via miR-150-5p and regulates ZEB1 expression.

Comparative analysis of three serum-free light-chain detection systems in the diagnosis of multiple myeloma.

OBJECTIVE: To observe the comparability of serum-free light chain (sFLC) detected by Beckman, Siemens, and Binding Site. METHODS: In this study, 110 patients who were diagnosed with multiple myeloma in State Key Laboratory of Experimental Hematology from November 2019 to August 2020. According to the instructions of equipment and reagent manufacturers, three detection systems (Binding Site, Beckman, and Siemens) were used to detect the serum-free light chain of selected analysis samples. Meanwhile, sFLC test results of the three instruments were compared. According to EP9-A3 guide, correlation and consistency were conducted by Bland-Altman and Passing-Bablok regression. RESULTS: The free kappa light-chain serum samples and the free lambda light-chain samples were quantitatively analyzed by the three systems. Binding Site, Beckman, and Siemens free light-chain detection results were as follows: FLC-κ: 42.23 (3.73, 423), 34.55 (6.5, 194), 39.85 (3.73, 423); FLC-λ: 31.46 (1.39, 8180.42), 32 (4.3, 275), 37.65 (2.28, 526); rFLC (FLC-κ/FLC-λ): 1.21 (0, 131.28), 0.96 (0.02, 43.49), 1.04 (0.04, 40.29). The Kappa valuation of FLC-κ between Beckman and Binding Site is 0.97, Kappa valuation of FLC-λ between Beckman and Binding Site is 0.96, Kappa valuation of rFLC between Beckman and Binding Site is 0.97. The Kappa valuation of FLC-κ between Siemens and Binding Site is 0.96, Kappa valuation of FLC-λ between Siemens and Binding Site is 0.88, Kappa valuation of rFLC between Siemens and Binding Site is 0.94. CONCLUSION: All of the three systems can meet the needs of diagnosis and treatment in clinical application. These analyses showed a very good concordance between Beckman, Siemens, and Binding Site.

[Two Serum Free Light Chain Detection Systems in the Diagnosis of Multiple Myeloma].

OBJECTIVE: To investigate the comparability of the Freelite, Binding Site, Beckman and N Latex FLC, Siemens in the detection of serum free light chain (sFLC) . METHODS: Fifty newly diagnosed multiple myeloma (MM) patients in Tianjin Institute of Blood Research from November 2019 to February 2020 were enrolled. The two systems (Freelite, Binding Site, Beckman and N Latex FLC, Siemens) were used to detect the sFLC of the samples. Outlier detection was performed by ESD method, methodological comparison and deviation assessment were performed by Passing-Bablok regression and Bland-Altman regression. RESULTS: Both the systems could quantitatively analyze free kappa light chain serum samples and free lambda light chain samples. Freelite, Binding Site, Beckman and N Latex FLC, Siemens free light chain test showed FLC-κ：36.5 (6.5, 194), 40.5 (6.94, 288), FLC-λ: 30.1 (4.3, 170.5), 35.1 (2.28, 526), rFLC (FLC-κ/ FLC-λ) : 0.82 (0.05, 43.25), 1.03 (0.03, 32.04), dFLC (｜FLC-κ- FLC-λ｜) : -5.8 (-161.97, 183.7), 1.1 (-505.1, 279.01), which existed no outliers. There were systematic differences, and the deviation level was not within the clinically acceptable range. CONCLUSION: Both the systems can meet the needs of clinical diagnosis and treatment, but there is a significant deviation between the two systems, the results are not comparable, and should be analyzed separately. In particular, the same system should be selected for monitoring the prognosis of MM.

Extraction, modification, and property characterization of dietary fiber from Agrocybe cylindracea.

Dietary fiber (DF) has gained a great attention owing to its potential health benefits. Agrocybe cylindracea is an edible fungus with high protein and low fat contents, which is also an enriched source of DF. However, limited study has been conducted on optimizing the conditions of A. cylindracea-derived DF extraction and modification as well as characterizing its properties. In this study, ultrasound-assisted enzymatic method for DF extraction was optimized as the following conditions: liquid material ratio of 29 ml/g, α-amylase concentration of 1.50%, protamex concentration of 1.20%, and ultrasonic power of 150 W, which improved the DF extraction yield to 37.70%. Moreover, high temperature modification (HTM) and cellulase modification (CEM) were applied to modify A. cylindracea-derived DF. The results showed that HTM had more potential capacity in converting insoluble DF into soluble DF, and DF with HTM exhibited more advantages in its physicochemical properties than DF with CEM. The DF with both HTM and CEM showed antioxidant activities, reflected by the increased reducing power as well as DPPH radical, hydroxyl radical, and ABTS+ scavenging capabilities in vitro. These findings could offer a reference for the extraction, modification, and characterizing various properties of DF from A. cylindracea, which would establish the foundation for the comprehensive application of fungi-derived DF.

Influence of patient characteristics on chimeric antigen receptor T cell therapy in B-cell acute lymphoblastic leukemia.

CD19-specific chimeric antigen receptor T cell (CD19 CAR T) therapy has shown high remission rates in patients with refractory/relapsed B-cell acute lymphoblastic leukemia (r/r B-ALL). However, the long-term outcome and the factors that influence the efficacy need further exploration. Here we report the outcome of 51 r/r B-ALL patients from a non-randomized, Phase II clinical trial (ClinicalTrials.gov number: NCT02735291). The primary outcome shows that the overall remission rate (complete remission with or without incomplete hematologic recovery) is 80.9%. The secondary outcome reveals that the overall survival (OS) and relapse-free survival (RFS) rates at 1 year are 53.0 and 45.0%, respectively. The incidence of grade 4 adverse reactions is 6.4%. The trial meets pre-specified endpoints. Further analysis shows that patients with extramedullary diseases (EMDs) other than central nervous system (CNS) involvement have the lowest remission rate (28.6%). The OS and RFS in patients with any subtype of EMDs, higher Tregs, or high-risk genetic factors are all significantly lower than that in their corresponding control cohorts. EMDs and higher Tregs are independent high-risk factors respectively for poor OS and RFS. Thus, these patient characteristics may hinder the efficacy of CAR T therapy.

miR-301a Suppression within Fibroblasts Limits the Progression of Fibrosis through the TSC1/mTOR Pathway.

Pulmonary fibrosis has been characterized by abnormal proliferation of fibroblasts and massive deposition of the extracellular matrix, which results from a complex interplay of chronic injury and inflammatory responses. MicroRNA-301a (miR-301a) is activated by multiple inflammatory stimulators, contributing to multiple tumorigenesis and autoimmune diseases. This study showed that miR-301a was overexpressed in a bleomycin-induced murine model of pulmonary fibrosis and patients with idiopathic pulmonary fibrosis (IPF). In addition, miR-301a was activated by transforming growth factor ß (TGF-ß) and interleukin 6 (IL-6) in normal and IPF fibroblasts, which was markedly reversed by the signal transducer and activator of transcription 3 (STAT3) inhibitor. The genetic ablation of miR-301a in mice reduced bleomycin-induced lung fibrosis, and the downregulation of miR-301a restrained proliferation and activation of fibroblasts. Furthermore, this study demonstrated that TSC1 was a functional target of miR-301a in fibroblasts, and the negative regulation of TSC1 by miR-301a promoted the severity of pulmonary fibrosis through the mammalian target of rapamycin (mTOR) signaling pathway. The blocking of miR-301a by the intravenous injection of antagomiR-301a inhibited the proliferation of fibroblasts and the structural destruction of lung tissues in the bleomycin-induced lung fibrosis mouse model. The findings revealed the crucial role of the miR-301a/TSC1/mTOR axis in the pathogenesis of pulmonary fibrosis, suggesting that miR-301a might serve as a potential therapeutic target.

Vitexin improves neuron apoptosis and memory impairment induced by isoflurane via regulation of miR-409 expression.

BACKGROUND: Anesthetics, such as isoflurane, sevoflurane, ketamine, and desflurane, are commonly used in clinics. Specifically, isoflurane is one of the most commonly used inhalational anesthetics, which can be used in surgery patients of all ages, including children. OBJECTIVES: The aim of the study was to investigate the mechanisms of vitexin against isoflurane-induced neurotoxicity. MATERIAL AND METHODS: Reference memory testing was performed for 5 days (4 trials, 2 per day) before anesthesia. Reversal testing was performed on the 3rd day after anesthesia. The cell viability and apoptosis of PC-12 cells were detected using MTT and TUNEL assays, respectively. Enzyme-linked immunosorbent assay (ELISA) kits were used to measure serum tumor necrosis factor α (TNF­α), interleukin 6 (IL­6), glutathione (GSH), and superoxide dismutase (SOD) concentrations. The concentration of reactive oxygen species (ROS) was detected using ROS measurement. Expression of miR-409 was determined using quantitative reverse-transcription polymerase chain reaction (qPT-PCR). Protein expression levels were detected using western blotting. RESULTS: Rats treated with isoflurane showed significant increases in the escape latency periods (ELP) and the apoptosis of hippocampus neuron cells; this effect was reversed by 3 mg/kg or 10 mg/kg of vitexin (p < 0.05). Further testing showed that isoflurane could significantly decrease the cell viability and increase the apoptosis of PC-12, the expression of inflammatory cytokines (TNF­α and IL­6) and ROS (p < 0.05). However, these results were reversed by 10/100 µM of vitexin. In addition, vitexin could significantly increase the expression of miR-409 (p < 0.05). Further studies showed that overexpression of miR-409 could significantly promote the effect of vitexin on isoflurane-induced neurotoxicity (p < 0.05). Finally, overexpression miR-409 could significantly increase the expression of p-AMPK/t-AMPK and p-GSK3ß/t-GSK3ß. CONCLUSIONS: Vitexin has protective effects against isoflurane-induced neurotoxicity by targeting miR-409 and the AMPK/GSK3ß pathway.

Enhanced lincomycin production by co-overexpression of metK1 and metK2 in Streptomyces lincolnensis.

Streptomyces lincolnensis is generally utilized for the production of lincomycin A (Lin-A), a clinically useful antibiotic to treat Gram-positive bacterial infections. Three methylation steps, catalyzed by three different S-adenosylmethionine (SAM)-dependent methyltransferases, are required in the biosynthesis of Lin-A, and thus highlight the significance of methyl group supply in lincomycin production. In this study, we demonstrate that externally supplemented SAM cannot be taken in by cells and therefore does not enhance Lin-A production. Furthermore, bioinformatics and in vitro enzymatic assays revealed there exist two SAM synthetase homologs, MetK1 (SLCG_1651) and MetK2 (SLCG_3830) in S. lincolnensis that could convert L-methionine into SAM in the presence of ATP. Even though we attempted to inactivate metK1 and metK2, only metK2 was deleted in S. lincolnensis LCGL, named as ΔmetK2. Following a reduction of the intracellular SAM concentration, ΔmetK2 mutant exhibited a significant decrease of Lin-A in comparison to its parental strain. Individual overexpression of metK1 or metK2 in S. lincolnensis LCGL either elevated the amount of intracellular SAM, concomitant with 15% and 22% increase in Lin-A production, respectively. qRT-PCR assays showed that overexpression of either metK1 or metK2 increased the transcription of lincomycin biosynthetic genes lmbA and lmbR, and regulatory gene lmbU, indicating SAM may also function as a transcriptional activator. When metK1 and metK2 were co-expressed, Lin-A production was increased by 27% in LCGL, while by 17% in a high-yield strain LA219X.

Effects of glenohumeral internal rotation deficit on baseball pitching among pitchers of different ages.

BACKGROUND: Previous studies have reported that glenohumeral internal rotation deficit (GIRD) may increase the risk of shoulder injury. However, the effects of GIRD on baseball pitching among pitchers of different age groups are still unclear. METHODS: The study participants were 24 high school and 24 university pitchers. For each age group, the pitchers were evenly divided into a GIRD group and a normal group. The pitching motion of each participant was captured using a motion analysis system at a sampling frequency of 300 Hz. The kinematics and kinetics of the throwing shoulder and trunk were quantified, and statistical differences between the groups were examined by 2-sample t tests. RESULTS: For both age groups, significant differences were observed in shoulder external rotations of the GIRD and normal groups. Compared with the university pitchers in the normal group, the university pitchers with GIRD exhibited a greater shoulder loading and did more internal rotation work in the acceleration phase. The high school pitchers with GIRD showed a larger trunk tilt and less trunk rotation than the university pitchers with GIRD. However, the university pitchers with GIRD exhibited a larger shoulder posterior force and horizontal adduction torque than the high school pitchers with GIRD. CONCLUSION: Pitchers with GIRD do change their pitching motions, and the greater resulting shoulder joint loading predisposes them to a greater risk of shoulder injury, especially among university pitchers.

Iodine-131 induces apoptosis in human cardiac muscle cells through the p53/Bax/caspase-3 and PIDD/caspase-2/ t­BID/cytochrome c/caspase-3 signaling pathway.

The aim of this study was to elucidate the effects of iodine-131 on the induction of apoptosis in human cardiac muscle cells and the underlying molecular mechanisms. We found that iodine-131 reduced cell proliferation, induced apoptosis, induced p53, PIDD, t-BID (mitochondria) protein expression, suppressed cytochrome c (mitochondria) protein expression, and increased Bax protein expression, and promoted caspase-2, -3 and -9 expression levels in human cardiac muscle cells. Meanwhile, si-p53 inhibited the effects of iodine-131 on the reduction in cell proliferation and induction of apoptosis in human cardiac muscle cells through regulation of Bax/cytochrome c/caspase-3 and PIDD/caspase­2/t-BID/cytochrome c/caspase-3 signaling pathway. After si-Bax reduced the effects of iodine-131, it reduced cell proliferation and induced apoptosis in human cardiac muscle cells through the cytochrome c/caspase-3 signaling pathway. However, si-caspase-2 also reduced the effects of iodine-131 on the reduction of cell proliferation and induction of apoptosis in human cardiac muscle cells through the t-BID/cytochrome c/caspase-3 signaling pathway. These findings demonstrated that iodine-131 induces apoptosis in human cardiac muscle cells through the p53/Bax/caspase-3 and PIDD/caspase-2/t-BID/cytochrome c/caspase-3 signaling pathway.

Characterization of an Lrp/AsnC family regulator SCO3361, controlling actinorhodin production and morphological development in Streptomyces coelicolor.

Lrp/AsnC family regulators have been found in many bacteria as crucial regulators controlling diverse cellular processes. By genomic alignment, we found that SCO3361, an Lrp/AsnC family protein from Streptomyces coelicolor, shared the highest similarity to the SACE_Lrp from Saccharopolyspora erythraea. Deletion of SCO3361 led to dramatic reduction in actinorhodin (Act) production and delay in aerial mycelium formation and sporulation on solid media. Dissection of the mechanism underlying the function of SCO3361 in Act production revealed that it altered the transcription of the cluster-situated regulator gene actII-ORF4 by directly binding to its promoter. SCO3361 was an auto-regulator and simultaneously activated the transcription of its adjacent divergently transcribed gene SCO3362. SCO3361 affected aerial hyphae formation and sporulation of S. coelicolor by activating the expression of amfC, whiB, and ssgB. Phenylalanine and cysteine were identified as the effector molecules of SCO3361, with phenylalanine reducing the binding affinity, whereas cysteine increasing it. Moreover, interactional regulation between SCO3361 and SACE_Lrp was discovered for binding to each other's target gene promoter in this work. Our findings indicate that SCO3361 functions as a pleiotropic regulator controlling secondary metabolism and morphological development in S. coelicolor.