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Abstract Neuronal apoptosis has been well-recognized as a critical mediator in the pathogenesis of depressive disorders. Tissue kallikrein-related peptidase 8 (KLK8), a trypsin-like serine protease, has been implicated in the pathogenesis of several psychiatric disorders. The present study aimed to explore the potential function of KLK8 in hippocampal neuronal cell apoptosis associated with depressive disorders in rodent models of chronic unpredictable mild stress (CUMS)-induced depression. It was found that depression-like behavior in CUMS-induced mice was associated with hippocampal KLK8 upregulation. Transgenic overexpression of KLK8 exacerbated, whereas KLK8 deficiency attenuated CUMS-induced depression-like behaviors and hippocampal neuronal apoptosis. In HT22 murine hippocampal neuronal cells and primary hippocampal neurons, adenovirus-mediated overexpression of KLK8 (Ad-KLK8) was sufficient to induce neuron apoptosis. Mechanistically, it was identified that the neural cell adhesion molecule 1 (NCAM1) may associate with KLK8 in hippocampal neurons as KLK8 proteolytically cleaved the NCAM1 extracellular domain. Immunofluorescent staining exhibited decreased NCAM1 in hippocampal sections obtained from mice or rats exposed to CUMS. Transgenic overexpression of KLK8 exacerbated, whereas KLK8 deficiency largely prevented CUMS-induced loss of NCAM1 in the hippocampus. Both adenovirus-mediated overexpression of NCAM1 and NCAM1 mimetic peptide rescued KLK8-overexpressed neuron cells from apoptosis. Collectively, this study identified a new pro-apoptotic mechanism in the hippocampus during the pathogenesis of CUMS-induced depression via the upregulation of KLK8, and raised the possibility of KLK8 as a potential therapeutic target for depression.

AIM: To investigate the expression and correlation of C1q/tumor necrosis factor related protein 9(CTRP9)levels in the serum of patients with different stages of diabetic retinopathy(DR)and diabetic macular edema(DME).METHODS: A total of 135 patients with type 2 diabetes who were admitted to Gansu Provincial Hospital from April 2021 to April 2022 were selected as the experimental group. According to the results of non-mydriatic fundus photography, they were divided into non-DR(NDR)group(n=45), non-proliferative DR(NPDR)group(n=45), proliferative DR(PDR)group(n=45); according to the results of optical coherence tomography, DR patients were divided into DME group(n=51), non-DME group(n=39). In addition, other 45 healthy subjects who matched the age and sex of the experimental group were selected as normal control group. The clinical data and biochemical index test results of subjects in each group were recorded and compared, the correlation between serum CTRP9 level and other biochemical indexes was analyzed, and the risk factors affecting the occurrence of DR and DME were explored.RESULTS: There were significant differences in serum CTRP9 levels among subjects in normal control group, NDR group, NPDR group and PDR group(P NDR group > NPDR group > PDR group. There was significant difference in serum CTRP9 level between DME group and non-DME group(P DME group. Spearman rank correlation analysis showed that the level of serum CTRP9 in DR patients was negatively correlated with the course of diabetes(rs=-0.251, P

AIM: To prepare the model of diabetic retinopathy(DR)on the basis of the model of type 2 diabetes mellitus(T2DM)in rats, observe the preventive and protective effect of metformin on DR in T2DM rats and the effect on serum Cystatin C(Cys C), and discuss its mechanism.METHODS: A total of 120 male SD rats were selected and 30 rats were randomly divided into blank control group A(10 rats), T2DM group(10 rats)and metformin intervention group A(10 rats). The remaining 90 rats were randomly divided into three groups: blank control group B(30 rats), DR group(30 rats)and metformin intervention group B(30 rats). Except for blank control groups A and B, T2DM models were constructed in all other groups. After modeling, rats in metformin intervention group A were given metformin by gavage, and rats in blank control group A and T2DM group were given normal saline by gavage. After 3mo of intervention, fasting blood glucose(FBG)and fasting serum insulin(FINS)indexes of the three groups were measured respectively, and the homeostasis model assessment of insulin resistance(HOMA-IR)was calculated and analyzed; The serum Cys C, tumor necrosis factor-α(TNF-α), the levels of Interleukin-8(IL-8), vascular endothelial growth factor(VEGF)and reactive oxygen species(ROS)were measured, and we observed the retinal tissue and vascular morphology of rats by FFA, HE staining and transmission electron microscopy; rats in metformin intervention group B began to give metformin by gavage 3mo after the course of T2DM, and other rats were given normal saline. According to the length of intervention, 10 rats were taken from each group and observed at 4, 5 and 6mo respectively. Serum levels of Cys C, TNF-α, IL-8, VEGF and ROS were measured in each group. We observed the retinal tissue and vascular morphology of rats by FFA, HE staining and transmission electron microscopy.RESULTS: The expressions of serum FBG, FINS, HOMA-IR, Cys C, TNF-α, IL-8, VEGF and ROS were statistically significant among the blank control group A, T2DM group and metformin intervention group A(P

Abstract Cancer-derived exosomal miRNAs are implicated in tumorigenesis and development of lung adenocarcinoma (LUAD). The objective of this study is to unravel the biological function of exosomal miR-197-3p in LUAD metastasis. qRT-PCR showed that elevated miR-197-3p in LUAD tissues was positively correlated with LUAD metastasis. CCK-8, tube formation, transwell and wound healing assays revealed that exosomal miR-197-3p from LUAD cells promoted the proliferation, angiogenesis and migration of HUVECs in vitro. LUAD cells-derived exosomal miR-197-3p also facilitated tumor growth and angiogenesis in LUAD cells-derived tumor xenograft model. TIMP2 and TIMP3 were identified as target genes of miR-197-3p in HUVECs by bioinformatics analysis and luciferase reporter assay. Functional studies illustrated that exosomal miR-197-3p promoted angiogenesis and migration via targeting TIMP2 and TIMP3 in HUVECs. In vivo data further supported that exosomal miR-197-3p promoted lung metastasis via TIMP2/3-mediated angiogenesis. In conclusion, LUAD cells-derived exosomal miR-197-3p conferred angiogenesis via targeting TIMP2/3 in LUAD metastasis.

Unfolded protein response (UPR) signaling and endoplasmic reticulum (ER) stress have been linked to pulmonary fibrosis. However, the relationship between UPR status and pulmonary function and prognosis in idiopathic pulmonary fibrosis (IPF) patients remains largely unknown. Through a series of bioinformatics analyses, we established a correlation between UPR status and pulmonary function in IPF patients. Furthermore, thrombospondin-1 (TSP-1) was identified as a potential biomarker for prognostic evaluation in IPF patients. By utilizing both bulk RNA profiling and single-cell RNA sequencing data, we demonstrated the upregulation of TSP-1 in lung fibroblasts during pulmonary fibrosis. Gene set enrichment analysis (GSEA) results indicated a positive association between TSP-1 expression and gene sets related to the reactive oxygen species (ROS) pathway in lung fibroblasts. TSP-1 overexpression alone induced mild ER stress and pulmonary fibrosis, and it even exacerbated bleomycin-induced ER stress and pulmonary fibrosis. Mechanistically, TSP-1 promoted ER stress and fibroblast activation through CD47-dependent ROS production. Treatment with either TSP-1 inhibitor or CD47 inhibitor significantly attenuated BLM-induced ER stress and pulmonary fibrosis. Collectively, these findings suggest that the elevation of TSP-1 during pulmonary fibrosis is not merely a biomarker but likely plays a pathogenic role in the fibrotic changes in the lung.

ObjectiveGestational diabetes mellitus (GDM) is a common glucose metabolism disease occurs in pregnancy that affects both maternal and neonatal health. Recently, increasing studies have attached importance to the relationship between growth differentiation factor 15 (GDF-15) and GDM, but the results were inconclusive. Therefore, we conducted a meta-analysis to examine the association between GDF-15 and GDM.Materials and methodsA systematical search was performed in Gene Expression Omnibus (GEO), PubMed and Google Scholar till Oct 27, 2022. We first calculated the mean and standard deviation of GDF-15 expression levels from the included eligible datasets and articles. Then, a meta-analysis was conducted to depict the difference in GDF-15 mRNA or GDF-15 protein expression between case and control groups by using conservative random effect model. Moreover, the potential publication bias was checked with the aid of Begg’s test and Egger’s test. Finally, sensitivity analyses were performed by changing the inclusion criteria.ResultsIn summary, 12 GEO datasets and 5 articles were enrolled in our study, including 789 GDM patients and 1202 non-GDM pregnant women. It was found that the expression levels of GDF-15 mRNA and GDF-15 protein in late pregnancy were significantly higher in GDM patients compared with non-GDM pregnant women, with the standard mean difference (SMD) and 95% confidence interval (95% CI) of 0.48 (0.14, 0.83) and 0.82 (0.32-1.33), respectively. Meanwhile, a slightly weakened association between GDF-15 protein levels and GDM was also observed in the middle pregnancy, with SMD (95% CI) of 0.53 (0.04-1.02).ConclusionIn all, our results suggested that the expression levels of GDF-15 were significantly higher in GDM patients compared with non-GDM pregnant women, especially in the late pregnancy.

Lysyl oxidase(LOX), an amino oxidase with copper binding site outside the cell, has been confirmed that may be involved in the pathogenesis of many diseases by a large number of studies. At present, although the potential mechanism of common variants of LOX in the development of disease is still unknown, but the potential role may be to affect the biological activity of LOX through tissue-specific alternative splicing or expression regulation.However, no matter under normal or pathological conditions, there is still little information about LOX in eye tissue yet.In recent years, it has been found that LOX may be involved in the occurrence and development of many eye diseases in related studies. Based on this, the review will summarize the research progress of eye diseases in which LOX is involved of recent years.

Objective To explore the feasibility and effect of electrocorticography (ECoG) for intraoperative mapping of motor cortex. Methods Eight patients with brain lesions near motor areas were operated under awake craniotomy from June 2017 to June 2019. The motor⁃related cortices were detected by intraoperative direct electrical stimulation (DES). The ECoG signals including μ rhythm and slow cortical potential (SCP) were collected intraoperatively and analyzed by wavelet analysis from patients at rest and hand⁃moving state (grasp and extension motion). The changes of event⁃related desynchronization (ERD) of different rhythms were analyzed by wavelet analysis before and after hand grasping. The threshold values of μ rhythm, SCP and different pattern recognition in combination were compared by using DES as "gold standard". The receiver operating characteristic (ROC) area under the curve (AUC) and the sensitivity and specificity were evaluated. Results When the threshold of ERD of the μ rhythm was chosen as 40%, the sensitivity and specificity of ECoG mapping of motor cortex were 81.08% (30/37) and 83.33% (25/30). The ERD threshold of the SCP was chosen as 1.6, the sensitivity and specificity were 83.78% (31/37) and 80% (24/30) respectively. Combined μ rhythm with SCP mapping revealed high sensitivity (97.29%, 36/37) and moderate specificity (80%, 24/30) as compared with DES when D mode (sensitive priority mode) was chosen, that was either μ rhythm or SCP positive was defined as the motor cortex. Conclusions The ECoG analysis has a higher sensitivity to locate the motor cortex without stimulating the brain. It is expected as a new adjuvant to DES for intraoperative mapping of eloquent cortex. DOI:10.3969/j.issn.1672⁃6731.2020.11.005

The rice mutant ossac4 (starch accumulating 4) was raised from seeds of the rice (Oryza sativa L.) indica maintainer line Xinong 1B treated with ethyl methanesulfonate. The distal and medial portions of the second leaf displayed premature senescence in the ossac4 mutant at the four-leaf stage. Physiological and biochemical analysis, and cytological examination revealed that the ossac4 mutant exhibited the premature leaf senescence phenotype. At the four-leaf stage, the leaves of the ossac4 mutant exhibited significantly increased contents of starch compared with those of the wild type (WT). Quantitative real-time PCR analysis showed that the expression levels of photosynthesis-associated genes were down-regulated and the expression levels of glucose metabolism-associated genes were abnormal. Genetic analysis indicated that the ossac4 mutation was controlled by a single recessive nuclear gene. The OsSAC4 gene was localized to a 322.7-kb interval between the simple-sequence repeat marker XYH11-90 and the single-nucleotide polymorphism marker SNP5300 on chromosome 11. The target interval contained 20 annotated genes. The present results demonstrated that ossac4 represents a novel starch accumulation and premature leaf senescence mutant, and lays the foundation for cloning and functional analysis of OsSAC4.

Objective: To assess the anti-inflammatory efficacy of ferruginol on dextran sulfate sodium (DSS) stimulated ulcerative colitis mice. Methods: Ulcerative colitis was induced in C57BL/6J mice by administering 2% of DSS through drinking water for 7 d. The mice in the treatment group were treated with DAA+50 mg/kg/day ferruginol orally. In the positive control group, sulfasalazine (50 mg/kg/day) was used alongside with DSS. After induction, the bodyweight, character of stool and feces occult blood were recorded daily, the disease activity index was calculated, and the colon length, colon weight, and spleen weight were recorded. The myeloperoxidase activity was assayed by spectrophotometry. Interleukin (IL)-6, IL-1β, and tumor necrosis factor-a were determined by ELISA method, and nuclear factor-κB, cyclooxygenase-2, matrix metalloproteinases-9, and inducible nitric oxide synthase by Western blotting assays. Results: Ferruginol significantly increased the bodyweight, colon weight, colon length, and decreased disease activity index and spleen weight. It exhibited anti-inflammatory activity against DSS induced ulcerative colitis in mice by reducing the activities of myeloperoxidase, tumor necrosis factor-α, nuclear factor-κβ, IL-1β, cyclooxygenase-2, matrix metalloproteinases-9, IL-6, and inducible nitric oxide synthase. Conclusions: Ferruginol could be used to treat ulcerative colitis by attenuating the inflammation in colon cells and maintaining colonic mucosal barrier integrity.

This experiment was designed to evaluate the potential for dietary alfalfa saponins (AS) to influence cholesterol (Ch) concentrations and the mRNA expressions of the ATP-binding cassette (ABC) that concludes transporters G5 (ABCG5) and G8 (ABCG8) in the livers of laying hens. Hundred and fifty laying hens (27 weeks old) were divided into five treatment groups randomly. Five replicates were assigned to each treatment group consisting of six birds per replicate. The same basic diet was used for all groups with supplementation of 0 (control), 60, 120, 240, and 480 mg/kg of AS, which was given ad libitum to hens for 60 days. The results showed that AS in the diets of laying hens could effectively reduce egg Ch concentrations, and increase the excretion of bile acid by the liver with the expression levels of ABCG5 and ABCG8 improving. Our study found that dietary AS could effect cholesterol metabolism by up-regulating the expression of G5 and G8 in hens, and that AS is a potential agent for reducing Ch concentrations and 120 mg/kg of AS in the diet was suitable and effective for laying hens.

Background: Salidroside (SDS) is the main effective ingredient of Rhodiola rosea L with a variety of pharmacologic properties. We aim to investigate the effects of SDS on ventilation induced lung injury (VILI) and explore the possible underlying molecular mechanism. Methods: Lung injury was induced in male ICR mice via mechanical ventilation (30 ml/kg) for 4h. The mice were divided in four groups:(1) Control group; (2) Ventilation group; (3) SDS group; (4) Ventilation with SDS group. SDS (50 mg/kg) was injected intraperitoneally 1h before operation. Mouse lung vascular endothelial cells (MLVECs) were subjected to cyclic stretch for 4h. Results: It was found that SDS attenuated VILI as shown in HE staining, cell count and protein content levels in BAL fluid, W/D and Evans blue dye leakage into the lung tissue. SDS treatment inhibited the activation of NLRP3 inflammasome and subsequent caspase-1 cleavage as well as interleukin (IL)-1β secretion both in vivo and in vitro. Moreover, SDS administration up-regulated SIRT1 expression. Importantly, knockdown of SIRT1 reversed the inhibitory effect of SDS on NLRP3 inflammasome activation. Conclusions: Taken together, these findings indicate that SDS may confer protection against ventilation induced lung injury via SIRT1-de-pendent inhibition of NLRP3 inflammasome activation.

Abstract Background Chemotherapy-induced nausea and vomiting adversely affects the quality of life of patients who receive chemotherapy via intravenous infusion or transcatheter arterial chemoembolization (TACE). This study aimed to investigate the clinical effects of transcutaneous electrical acupoint stimulation (TEAS) on nausea and vomiting after TACE. Methods A total of 142 patients who received TACE with cisplatin for primary or metastatic liver cancer were assigned to the active-acupuncture (n = 72) or placebo-acupuncture (n = 70) groups using a covariate-adaptive randomization at a ratio of 1:1. The acupoints Hegu (LI4), Neiguan (P6), and Zusanli (ST36) were stimulated twice daily for 6 days. The effects of TEAS on nausea and vomiting were assessed by using occurrence rate and severity of these symptoms. Anorexia scale and M. D. Anderson Symptom Inventory (MDASI) scores were secondary endpoints and were used to assess the effect of TEAS on patient appetite and quality of life. The safety of the treatments was also monitored. Results Between the two groups, the differences in occurrence rates and severities of nausea and vomiting after TACE were not significant (all P > 0.05). From the second day after TACE, anorexia scores were significantly lower in the active-acupuncture group than in the placebo-acupuncture group and continued to decrease over time with treatment (all P values less than 0.01). On days 0, 1, and 2, the mean MDASI scores for the active-acupuncture group were slightly lower than those for the placebo-acupuncture group, but the differences were not statistically significant (all P > 0.05). No significant differences were found between the two groups in the occurrence rate of any adverse event (P > 0.05). Conclusion TEAS appears to be a safe and effective therapy to relieve patients’ gastrointestinal discomfort after chemotherapy. Trial registration NCT01895010. Registered 21 June 2013

Objective: To study the expression quantity of serum miR-146a and miR-146b in patients with acute cerebral infarction before and after the intervention of rosuvastatin and its correlation with toll-like receptor 2 (TLR2) and TLR4 signaling pathways. Methods: A total of 65 patients with acute cerebral infarction treated in our hospital from December 2015 to August 2016 were selected for prospective study. They were treated with lipid-lowering rosuvastatin, and peripheral blood samples were collected at 8th week before and after treatment, respectively. Serum was separated and expression quantity of miR-146a and miR-146b and contents of TNF-α, interleukin (IL)-1β, IL-6 and IL-17 were determined. Peripheral blood mononuclear cells were isolated and fluorescence intensities of TLR2, TLR4, myeloid differentiation primary response gene 88 (MyD88), interleukin-1 receptor-associated kinase 1 (IRAK-1) and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-kB) were measured. >Results: At 8th week of intervention of rosuvastatin, expression quantity of serum miR-146a [(0.762 ± 0.092) vs. (0.346 ± 0.053)] and miR-146b [(0.714 ± 0.088) vs. (0.317 ± 0.047)] in patients with acute cerebral infarction was significantly higher than those before the intervention. Fluorescence intensities of peripheral blood mononuclear cells such as TLR2 [(10.34 ± 1.27) vs. (16.94 ± 1.94)], TLR4 [(11.37 ± 1.54) vs. (24.35 ± 3.26)], IRAK [(9.34 ± 0.92) vs. (15.32 ± 1.82)], MyD88 [(4.42 ± 0.56) vs. (9.41 ± 1.03)] and NF-kB [(6.65 ± 0.78) vs. (13.49 ± 1.76)] and contents of inflammatory factors such as TNF-α [(64.26 ± 8.29) μg/L vs. (106.39 ± 13.84) μg/L], IL-1β [(37.91 ± 5.24) μg/L vs. (64.23 ± 8.33) μg/L], IL-6 [(34.28 ± 4.85) ng/L vs. (82.46 ± 11.97) ng/L] and IL-17 [(56.75 ± 7.49) ng/L vs. (98.31 ± 11.36) ng/ L] of serum were all significantly lower than those before the intervention. Expression quantity of serum miR-146a and miR-146b had a negative correlation with fluorescence intensities of TLR2, TLR4, IRAK, MyD88 and NF-kB. Fluorescence intensities of TLR2 and TLR4 in peripheral blood mononuclear cells had a positive correlation with contents of TNF-α, IL-1β, IL-6 and IL-17 in serum. >Conclusions: Treatment with rosuvastatin can up-regulate the expression quantity of serum miR-146a and miR-146b in patients with acute cerebral infarction and further inhibit the secretion of IRAK, MyD88, NF-kB, TNF-α, IL-1β, IL-6 and IL-17 mediated by TLR2 and TLR4.

Background: Hydrogen sulfide (H2S), known as the third endogenous gaseous transmitter, has received increasing attention because of its diverse effects, including angiogenesis, vascular relaxation and myocardial protection.We aimed to investigate the role of H2S in oxidative/nitrative stress and inflammation in acute lung injury (ALI) induced by endotoxemia. Methods: Male ICR mice were divided in six groups: (1) Control group; (2) GYY4137treatment group; (3) L-NAME treatment group; (4) lipopolysaccharide (LPS) treatment group; (5) LPS with GYY4137 treatment group; and (6) LPS with L-NAME treatment group. The lungs were analysed by histology, NO production in the mouse lungs determined by modified Griess (Sigma-Aldrich) reaction, cytokine levels utilizing commercialkits, and protein abundance by Western blotting. Results: GYY4137, a slowly-releasing H2S donor, improved the histopathological changes in the lungs of endotoxemic mice. Treatment with NG-nitro-L-arginine methyl ester (L-NAME), a nitric oxide synthase (NOS) inhibitor, increased anti-oxidant biomarkers such as thetotal antioxidant capacity (T-AOC) and theactivities of catalase (CAT) and superoxide dismutase (SOD) but decreased a marker of peroxynitrite (ONOO-) action and 3-nitrotyrosine (3-NT) in endotoxemic lung. L-NAME administration also suppressed inflammation in endotoxemic lung, as evidenced by the decreased pulmonary levels of interleukin (IL)-6, IL-8, and myeloperoxidase (MPO) and the increased level of anti-inflammatory cytokine IL-10. GYY4137 treatment reversed endotoxin-induced oxidative/nitrative stress, as evidenced by a decrease in malondialdehyde (MDA), hydrogenperoxide (H2O2) and 3-NT and an increase in the antioxidant biomarker ratio of reduced/oxidized glutathione(GSH/GSSG ratio) and T-AOC, CAT and SOD activity. GYY4137 also attenuated endotoxin-induced lung inflammation. Moreover, treatment with GYY4137 inhibited inducible NOS (iNOS) expression and nitric oxide (NO) production in the endotoxemia lung. Conclusions: GYY4137 conferred protection against acute endotoxemia-associated lung injury, which may have beendue to the anti-oxidant, anti-nitrative and anti-inflammatory properties of GYY4137. The present findings warrant further exploration of the clinical applicability of H2S in the prevention and treatment of ALI.

Objective: To analyze the serum levels of miR-126, miR-146a and its relationship with infarction area, severity of disease and inflammatory reaction degree in patients with acute cerebral infarction (ACI). Methods: A total of 75 cases with ACI treated in our hospital from April 2014 to October 2015 and 80 healthy cases were respectively selected as ACI group and control group for retrospective study. Patients' clinical data were collected, and the serum levels of miR-126, miR-146a, tumor necrosis factor-a (TNF-a), interleukin-1b (IL-1b) and IL-6 were detected. Results: Serum contents of miR-126 (0.286 ± 0.078 vs. 1.000 ± 0.169) and miR-146a (0.337 ± 0.084 vs. 1.000 ± 0.158) in patients of ACI group were significantly lower than those of control group. Contents of IL-1b [(68.4 ± 10.3) vs. (22.7 ± 5.8) ng/L], TNFa [(126.9 ± 22.4) vs. (49.6 ± 8.4) ng/L] and IL-6 [(89.3 ± 14.7) vs. (34.8 ± 5.9) ng/L] were obviously higher than those of control group. The bigger the infarction area was, the more severer the degree of nerve defect could be. The lower the serum levels of miR-126, miR-146a were, the higher the levels of TNF-a, IL-1b, IL-6 could be. Levels of miR-126 and miR-146a were negative correlation with levels of TNF-a, IL-1b and IL-6. Conclusions: An abnormal decrease in serum levels of miR-126 and miR-146a in patients with ACI was closely related to the severity of disease. Through regulating the generation of inflammatory factors TNF-a, IL-1b and IL-6, miR-126 and miR-146a may get involved in the changes of cerebral infarction condition.

The nuclear-encoded light-harvesting chlorophyll a/b-binding proteins (LHCPs) are specifically translocated from the stroma into the thylakoid membrane through the chloroplast signal recognition particle (cpSRP) pathway. The cpSRP is composed of a cpSRP43 protein and a cpSRP54 protein, and it forms a soluble transit complex with LHCP in the chloroplast stroma. Here, we identified the YGL9 gene that is predicted to encode the probable rice cpSRP43 protein from a rice yellow-green leaf mutant. A phylogenetic tree showed that an important conserved protein family, cpSRP43, is present in almost all green photosynthetic organisms such as higher plants and green algae. Sequence analysis showed that YGL9 comprises a chloroplast transit peptide, three chromodomains and four ankyrin repeats, and the chromodomains and ankyrin repeats are probably involved in protein-protein interactions. Subcellular localization showed that YGL9 is localized in the chloroplast. Expression pattern analysis indicated that YGL9 is mainly expressed in green leaf sheaths and leaves. Quantitative real-time PCR analysis showed that the expression levels of genes associated with pigment metabolism, chloroplast development and photosynthesis were distinctly affected in the ygl9 mutant. These results indicated that YGL9 is possibly involved in pigment metabolism, chloroplast development and photosynthesis in rice.

Oxidative stress plays a pivotal role in the initiation and progression of cardiac diseases. Estrogens have been demonstrated to exert pleiotropic cardioprotective effects, among which antioxidative stress is one of the key effects linking estrogens to cardioprotection. By using a microRNAs (miRs) microarray screening approach, we discovered an increase in miR-494, which is known to exert cardioprotective effects, in estrogen-treated cardiomyocytes. We hypothesized that the upregulation of miR-494 might contribute to estrogen-mediated cardioprotection against oxidative stress. We found that E2 stimulates miR-494 expression via ERα in both cardiomyocytes and the myocardium of female mice. The miR-494 inhibitor attenuated the protective effect of 17β-estradiol (E2) against oxidative stress-induced injury in cardiomyocytes. By contrast, the miR-494 mimic protected cardiomyocytes against oxidative stress-induced cardiomyocyte injury. Using real-time PCR, western blot and dual-luciferase reporter gene analyses, we identified nuclear factor kappa B (NF-κB) repressing factor (NKRF) as the miR-494 target in cardiomyocytes. E2 was found to inhibit NKRF, thus activating NF-κB through a miR-494-dependent mechanism. In addition, the protective effects of E2 and miR-494 against oxidative stress in cardiomyocytes were eliminated by the NF-κB inhibitor. In summary, this study demonstrates for the first time that estrogen inhibits NKRF expression through ERα-mediated upregulation of miR-494 in cardiomyocytes, leading to the activation of NF-κB, which in turn results in an increase in antioxidative defense. ERα-mediated upregulation of miR-494 may contribute to estrogen protection of cardiomyocytes against oxidative stress.

BACKGROUND: Hepatocellular carcinoma (HCC) can be diagnosed by noninvasive approaches with serum α-fetoprotein (AFP) levels >200 ng/ml and/or a radiological imaging study of tumor mass >2 cm in patients with chronic liver disease. Percutaneous fine needle aspiration (FNA) under ultrasound (US) guidance has a diagnostic specificity of 95% and is superior to radiological imaging studies. AIM: The aim of this study is to elucidate the effectiveness and complications of fine needle aspiration in a Chinese population with primary liver cancer and AFP levels ≤200 ng/ml. MATERIALS AND METHODS: A retrospective study was conducted over a period of 28 years. This selection period included patients with a suspected diagnosis of primary liver cancer whose AFP levels were ≤200 ng/ml and who underwent US-FNA. This data was then analyzed with cytomorphological features correlating with medical history, radiological imaging, AFP, and follow-up information. RESULTS: Of the 1,929 cases with AFP ≤200 mg/ml, 1,756 underwent FNA. Of these, 1,590 cases were determined malignant and the remaining 166 were determined benign. Further, 1,478 malignant cases were diagnosed by FNA alone, and of these, 1,138 were diagnosed as PLC. The sensitivity, specificity, positive predictive value, negative predictive value, and overall accuracy of the diagnoses were 92.96%, 100%, 100%, 59.71%, and 93.62% respectively. There was no significant difference in the sensitivity, specificity, PPV and NPV between the subgroups with tumor size

BackgroundNitrilase is an important member of the nitrilase superfamiliy. It has attracted substantial interest from academia and industry for its function of converting nitriles directly into the corresponding carboxylic acids in recent years. Thus nitrilase has played a crucial role in production of commercial carboxylic acids in chemical industry and detoxification of nitrile-contaminated wastes. However, conventional studies mainly focused on the bacterial nitrilase and the potential of fungal nitrilase has been far from being fully explored. Research on fungal nitrilase gene expression will advance our understanding for its biological function of fungal nitrilase in nitrile hydrolysis.Methodology/principal findingsA fungal nitrilase gene from Gibberella intermedia was cloned through reverse transcription-PCR. The open reading frame consisted of 963 bp and potentially encoded a protein of 320 amino acid residues with a theoretical molecular mass of 35.94 kDa. Furthermore, the catalytic triad (Glu-45, Lys-127, and Cys-162) was proposed and confirmed by site-directed mutagenesis. The encoding gene was expressed in Escherichia coli Rosetta-gami (DE3) and the recombinant protein with His(6)-tag was purified to electrophoretic homogeneity. The purified enzyme exhibited optimal activity at 45°C and pH 7.8. This nitrilase was specific towards aliphatic and aromatic nitriles. The kinetic parameters V(max) and K(m) for 3-cyanopyridine were determined to be 0.81 µmol/min·mg and 12.11 mM through Hanes-Woolf plot, respectively. 3-Cyanopyridine (100 mM) could be thoroughly hydrolyzed into nicotinic acid within 10 min using the recombinant strain with the release of about 3% nicotinamide and no substrate was detected.Conclusions/significanceIn the present study, a fungal nitrilase was cloned from the cDNA sequence of G. intermedia and successfully expressed in E. coli Rosetta-gami (DE3). The recombinant strain displayed good 3-cyanopyridine degradation efficiency and wide substrate spectrum. This fungal nitrilase might be a potential candidate for industrial applications in carboxylic acids production.

BackgroundHuman uterus undergoes distinct molecular and functional changes during pregnancy and parturition. Hydrogen sulfide (H(2)S) has recently been shown to play a key role in the control of smooth muscle tension. The role of endogenous H(2)S produced locally in the control of uterine contractility during labour is unknown.Methodology/principal findingsHuman myometrium biopsies were obtained from pregnant women undergoing cesarean section at term. Immunohistochemistry analysis showed that cystathionine-γ-lyase (CSE) and cystathionine-β-synthetase (CBS), the principle enzymes responsible for H(2)S generation, were mainly localized to smooth muscle cells of human pregnant myometrium. The mRNA and protein expression of CBS as well as H(2)S production rate were down-regulated in labouring tissues compared to nonlabouring tissues. Cumulative administration of L-cysteine (10(-7)-10(-2) mol/L), a precursor of H(2)S, caused a dose-dependent decrease in the amplitude of spontaneous contractions in nonlabouring and labouring myometrium strips. L-cysteine at high concentration (10(-3) mol/L) increased the frequency of spontaneous contractions and induced tonic contraction. These effects of L-cysteine were blocked by the inhibitors of CBS and CSE. Pre-treatment of myometrium strips with glibenclamide, an inhibitor of ATP-sensitive potassium (K(ATP)) channels, abolished the inhibitory effect of L-cysteine on spontaneous contraction amplitude. The effects of L-cysteine on the amplitude of spontaneous contractions and baseline muscle tone were less potent in labouring tissues than that in nonlabouring strips.Conclusion/significanceH(2)S generated by CSE and CBS locally exerts dual effects on the contractility of pregnant myometrium. Expression of H(2)S synthetic enzymes is down-regulated during labour, suggesting that H(2)S is one of the factors involved in the transition of pregnant uterus from quiescence to contractile state after onset of parturition.