Integrated omics analysis reveals the immunologic characteristics of cystic Peyer's patches in the cecum of Bactrian camels.

Bactrian camels have specific mucosa-associated lymphoid tissue (MALT) throughout the large intestine, with species-unique cystic Peyer's patches (PPS) as the main type of tissue. However, detailed information about the molecular characteristics of PPS remains unclear. This study applied a transcriptomic analysis, untargeted metabolomics, and 16S rDNA sequencing to compare the significant differences between PPS and the adjacent normal intestine tissues (NPPS) during the healthy stage of three young Bactrian camels. The results showed that samples from PPS could be easily differentiated from the NPPS samples based on gene expression profile, metabolites, and microbial composition, separately indicated using dimension reduction methods. A total of 7,568 up-regulated and 1,266 down-regulated differentially expressed genes (DEGs) were detected, and an enrichment analysis found 994 DEGs that participated in immune-related functions, and a co-occurance network analysis identified nine hub genes (BTK, P2RX7, Pax5, DSG1, PTPN2, DOCK11, TBX21, IL10, and HLA-DOB) during multiple immunologic processes. Further, PPS and NPPS both had a similar pattern of most compounds among all profiles of metabolites, and only 113 differentially expressed metabolites (DEMs) were identified, with 101 of these being down-regulated. Deoxycholic acid (DCA; VIP = 37.96, log2FC = -2.97, P = 0), cholic acid (CA; VIP = 13.10, log2FC = -2.10, P = 0.01), and lithocholic acid (LCA; VIP = 12.94, log2FC = -1.63, P = 0.01) were the highest contributors to the significant dissimilarities between groups. PPS had significantly lower species richness (Chao1), while Firmicutes (35.92% ± 19.39%), Bacteroidetes (31.73% ± 6.24%), and Proteobacteria (13.96% ± 16.21%) were the main phyla across all samples. The LEfSe analysis showed that Lysinibacillus, Rikenellaceae_RC9_gut_group, Candidatus_Stoquefichus, Mailhella, Alistipes, and Ruminococcaceae_UCG_005 were biomarkers of the NPPS group, while Escherichia_Shigella, Synergistes, Pyramidobacter, Odoribacter, Methanobrevibacter, Cloacibacillus, Fusobacterium, and Parabacteroides were significantly higher in the PPS group. In the Procrustes analysis, the transcriptome changes between groups showed no significant correlations with metabolites or microbial communities, whereas the alteration of metabolites significantly correlated with the alteration of the microbial community. In the co-occurrence network, seven DEMs (M403T65-neg, M329T119-neg, M309T38-neg, M277T42-2-neg, M473T27-neg, M747T38-1-pos, and M482t187-pos) and 14 genera (e.g., Akkermansia, Candidatus-Stoquefichus, Caproiciproducens, and Erysipelatoclostridium) clustered much more tightly, suggesting dense interactions. The results of this study provide new insights into the understanding of the immune microenvironment of the cystic PPS in the cecum of Bactrian camels.

Randomized Trial of First-Line Tyrosine Kinase Inhibitor With or Without Radiotherapy for Synchronous Oligometastatic EGFR-Mutated Non-Small Cell Lung Cancer.

BACKGROUND: Adding radiotherapy (RT) to systemic therapy improves progression-free survival (PFS) and overall survival (OS) in oligometastatic non-small cell lung cancer (NSCLC). Whether these findings translate to epidermal growth factor receptor (EGFR)-mutated NSCLC remains unknown. The SINDAS trial (NCT02893332) evaluated first-line tyrosine kinase inhibitor (TKI) therapy for EGFR-mutated synchronous oligometastatic NSCLC and randomized to upfront RT vs no RT; we now report the prespecified interim analysis at 68% accrual. METHODS: Inclusion criteria were biopsy-proven EGFR-mutated adenocarcinoma (per amplification refractory mutation system or next generation sequencing), with synchronous (newly diagnosed, treatment naïve) oligometastatic (≤5 metastases; ≤2 lesions in any one organ) NSCLC without brain metastases. All patients received a first-generation TKI (gefitinib, erlotinib, or icotinib), and randomization was between no RT vs RT (25-40 Gy in 5 fractions depending on tumor size and location) to all metastases and the primary tumor/involved regional lymphatics. The primary endpoint (intention to treat) was PFS. Secondary endpoints included OS and toxicities. All statistical tests were 2-sided. RESULTS: A total of 133 patients (n = 65 TKI only, n = 68 TKI with RT) were enrolled (2016-2019). The median follow-up was 23.6 months. The respective median PFS was 12.5 months vs 20.2 months (P < .001), and the median OS was 17.4 months vs 25.5 months (P < .001) for TKI only vs TKI with RT. Treatment yielded no grade 5 events and a 6% rate of symptomatic grade 3-4 pneumonitis in the TKI with RT arm. Based on the efficacy results of this prespecified interim analysis, the ethics committee recommended premature cessation of this trial. CONCLUSIONS: As compared with a first-line TKI alone, addition of upfront local therapy using RT statistically significantly improved PFS and OS for EGFR-mutated NSCLC.

Longitudinal changes in the hypothalamic-pituitary-adrenal axis and sympathetic nervous system are related to the prognosis of stroke.

Background and purpose: This study sought to improve methods to identify biomarkers in the neuroendocrine system related to stroke progression to improve the accuracy of traditional tools for evaluating stroke prognosis. Methods: Seventy-four stroke patients and 237 healthy controls were prospectively included. We measured urinary epinephrine (E), noradrenaline (NE), dopamine (DA) and cortisol (F) on days 1, 3, and 5 after stroke onset and plasma F, adrenocorticotropic hormone (ACTH), thyrotropin (TSH), prolactin (PRL), follicle-stimulating hormone (FSH), luteinizing hormone (LH) and growth hormone (GH). The correlation between these hormone levels and 90-day prognosis was analyzed, their value in assessing prognosis was compared with lesion volume and National Institutes of Health Stroke Scale (NIHSS) scores using receiver operating characteristic (ROC) curves, and their correlation with conventional clinical variables was assessed. Results: Levels of F, 24-h urinary free cortisol(UFC), E, NE, DA, and GH on days 1, 3, and 5 were significantly higher in stroke patients than in controls (P < 0.01), while ACTH and TSH decreased, gradually approaching normal within 5 days of onset. Levels of E, NE, F, and 24-h UFC were proportional to severity, and all gradually decreased within 5 days of onset in patients with a good prognosis and gradually increased or remained high in those with a poor prognosis. After adjustment for age, sex, NIHSS, or Glasgow Coma Scale (GCS) score, F > 13.6 µg/dL, ACTH > 22.02 pg/mL and NE > 123.5 µg/ 24 h were identified as risk factors for a poor prognosis 90 days after stroke (P < 0.05). The combination of F, ACTH, NE, white blood cell count (WBC), glucose (Glu), and hemoglobin (Hb) was significantly more accurate than lesion volume (AUC: 0.931 vs. 0.694 P = 0.019) and NIHSS score (AUC: 0.931 vs. 0.746 P = 0.034) in predicting poor prognosis of stroke 1 day after onset. Hormones and traditional clinical variables were correlated to varying degrees, with NE correlating most strongly with 24-h UFC (r = 0.54) and moderately positively with lesion volume (r = 0.40) and NIHSS score (r = 0.45). Conclusions: Stroke causes significant time-phased dynamic changes in the hypothalamic-pituitary-adrenal axis and sympathetic nervous system, and plasma F, ACTH, and urinary NE levels can be used to assess stroke severity and prognosis. Chinese clinical trial registry: Registration Number: ChiCTR1900024992. Registration Date: 2019/8/6.

Challenges for a Cancer Center in the Novel Coronavirus Pneumonia Epidemic.

INTRODUCTION: The novel Coronavirus disease 2019 pandemic is sweeping through China, posing the greatest ever threat to its public health and economy. As a tertiary cancer center in Southwest China, we formulated and implemented an anti-infection protocol to prevent the spread of Coronavirus disease 2019 in our department. METHODS: The anti-infection protocol divided patients into 3 categories, namely outpatients, inpatients, and patients receiving radiation therapy at our cancer center, and each category had a distinct anti-infection protocol to minimize the risk of Coronavirus disease 2019 transmission. In each category, the patients were classified into high-, intermediate-, and low-risk groups. Each risk group was managed differently. A survey of patient volume changes prior to and during the Coronavirus disease 2019 outbreak was performed. RESULTS: We carried out the anti-infection protocol at our cancer center during the Coronavirus disease 2019 outbreak. We found that the total volume of both outpatient visits and inpatient treatment declined significantly depending on the conditions of each group. Radiation therapy and palliative service had the lowest and highest volume reductions at 58.3% and 100%, respectively. The decline in outpatient volumes was higher than the decline in inpatient treatment services (78.8% vs 71.8%). There was no Coronavirus disease 2019 cross-infection at our center, or Coronavirus disease 2019-related injury or death. The anti-infection protocol measures continue to be taken at the hospital even today but they have been modified depending on the prevalent local conditions. CONCLUSIONS: Challenges from the Coronavirus disease 2019 pandemic remain in our community. The anti-infection protocol implemented at our cancer center has been effective in preventing cross-infection. Whether our anti-infection protocol experience can be applied to curb the spread of the infection in other parts of the world remains to be tested.

Detection of Asymptomatic Carotid Artery Stenosis in High-Risk Individuals of Stroke Using a Machine-Learning Algorithm.

Objective Asymptomatic carotid stenosis (ACS) is closely associated to the incidence of severe cerebrovascular diseases. Early identifying the individuals with ACS and its associated risk factors could be beneficial for primary prevention of stroke. This study aimed to investigate a machine-learning algorithm for the detection of ACS among high-risk population of stroke based on the associated risk factors.Methods A novel model of machine learning was utilized to screen the associated predictors of ACS based on 30 potential risk factors. The algorithm of this model adopted a random forest pattern based on the training data and then was verified using the testing data. All of the original data were retrieved from the China National Stroke Screening and Prevention Project (CNSSPP), including demographic, clinical and laboratory characteristics. The individuals with high risk of stroke were enrolled and randomly divided into a training group and a testing group at a ratio of 4:1. The identification of carotid stenosis by carotid artery duplex scans was set as the golden standard. The receiver operating characteristic (ROC) curve and the area under the curve (AUC) was used to evaluate the efficacy of the model in detecting ACS.Results Of 2841 high risk individual of stroke enrolled, 326 (11.6%) were diagnosed as ACS by ultrasonography. The top five risk factors contributing to ACS in this model were identified as family history of dyslipidemia, high level of low-density lipoprotein cholesterol (LDL-c), low level of high-density lipoprotein cholesterol (HDL-c), aging, and low body mass index (BMI). Their weights were 11.8%, 7.6%, 7.1%, 6.1%, and 6.1%, respectively. The total weight of the top 15 risk factors was 85.5%. The AUC values of the model for detecting ACS with training dataset and testing dataset were 0.927 and 0.888, respectively.Conclusions This study demonstrated that the machine-learning algorithm could be used to identify the risk factors for ACS among high risk population of stroke. Family history of dyslipidemia may be the most important risk factor for ACS. This model could be a suitable tool to optimize the clinical approach for the primary prevention of stroke.

Indirubin-3'-monoxime suppresses amyloid-beta-induced apoptosis by inhibiting tau hyperphosphorylation.

Indirubin-3'-monoxime is an effective inhibitor of cyclin-dependent protein kinases, and may play an obligate role in neuronal apoptosis in Alzheimer's disease. Here, we found that indirubin-3'-monoxime improved the morphology and increased the survival rate of SH-SY5Y cells exposed to amyloid-beta 25-35 (Aß25-35), and also suppressed apoptosis by reducing tau phosphorylation at Ser199 and Thr205. Furthermore, indirubin-3'-monoxime inhibited phosphorylation of glycogen synthase kinase-3ß (GSK-3ß). Our results suggest that indirubin-3'-monoxime reduced Aß25-35-induced apoptosis by suppressing tau hyperphosphorylation via a GSK-3ß-mediated mechanism. Indirubin-3'-monoxime is a promising drug candidate for Alzheimer's disease.

A clinical survey of pain in Parkinson's disease in Eastern China.

BACKGROUND: This is a case-control study to investigate the prevalence, characteristics, and risk factors of pain in patients with Parkinson's disease (PD). METHODS: A total of 200 PD patients from eastern China were enrolled in our study. Accordingly, 200 healthy elderly adults were recruited as controls. The characteristics of pain were collected by using the Visual Analog Scale, Brief Pain Inventory (BPI), SF-36 Bodily Pain Scale, Unified Parkinson's Disease Rating Scale, Hoehn-Yahr Scale (H-Y), Hamilton Depression Scale, and Leeds Assessment of Neuropathic Symptoms and Signs. RESULTS: Of the 200 PD patients, pain was complained by 106 patients (53%). According to the SF-36 Bodily Pain Scale, pain morbidity in PD patients was significantly higher than in the control group. The average pain during last 24 h measured by the BPI was 2.67. About 76% of PD patients were found to have one pain type, 21.7% were having two pain types, and 1.9% had three pain types. Further, 69.8% of these patients were presented with musculoskeletal pain, 4.7% with dystonic pain, 22.6% with radicular-neuropathic pain, 20.8% with central neuropathic pain, and 9.4% with akathisia pain. The onset age and depression were the most significant predictors of pain in PD patients (p < 0.05). However, there was no significant association between pain and gender, age, disease duration, or severity of the disease. Only 5.7% of PD patients with pain received treatment in this study. CONCLUSIONS: Pain is frequent and disabling, independent of demographic and clinical variables, and is significantly more common in PD patients.

[Novel hybrids of (phenylsulfonyl)furoxan and N-benzyl matrinol as anti-hepatocellular carcinoma agents].

N-Benzyl matrinol was obtained by hydrolysis, benzylation and reduction reaction from matrine. A series of hybrids (8a-8n) from (phenylsulfonyl)furoxan and N-benzyl matrinol were synthesized and biologically evaluated as anti-hepatocellular carcinoma agents. All target compounds were evaluated for anti-proliferative activity against human hepatocellular Bel-7402, SMMC-7721, Bel-7404, and HepG2 cells in vitro by MTT method. The results indicated that all of these compounds had potent anti-proliferative activity which were more potent than their parent compound and 5-FU, especially 8a-8h and 8j showed the strongest anti-HCC HepG2 cell activity with IC50 values of 0.12-0.93 µmol x L(-1).

Comparative Effectiveness of Risk-adapted Surveillance vs Retroperitoneal Lymph Node Dissection in Clinical Stage I Nonseminomatous Germ Cell Testicular Cancer: A Retrospective Follow-up Study of 81 Patients.

PURPOSE: To retrospective assess the potential predictors for relapse and create an effective clinical mode for surveillance after orchidectomy in clinical stage I non-seminomatous germ cell testicular tumors (CSI-NSGCTs). MATERIALS AND METHODS: We analyzed data for CSI-NSGCTs patients with non-lymphatic vascular invasion, %ECa < 50% (percentage of embryonal carcinoma < 50%), and negative or declining tumor markers to their half-life following orchidectomy (defined as low-risk patients); these patients were recruited from four Chinese centers between January 1999 and October 2013. Patients were divided into active surveillance group and retroperitoneal lymph node dissection (RPLND) group according to different therapeutic methods after radical orchidectomy was performed. The disease-free survival rates (DFSR) and overall survival rates (OSR) of the two groups were compared by Kaplan-Meier analysis. RESULTS: A total of 121 patients with CSI-NSGCT were collected from four centers, and 81 low-risk patients, including 54 with active surveillance and 27 with RPLND, were enrolled at last. The median follow-up duration was 66.2 (range 6-164) months in the RPLND group and 65.9 (range 8-179) months in the surveillance group. OSR was 100% in active surveillance and RPLND groups, and DFSR was 89.8% and 87.0%, respectively. No significant difference was observed between these two groups (X2=0.108, P=0.743). No significant difference was observed between the patients with a low percentage of embryonal carcinoma (<50%) and those without embryonal carcinoma (87.0% and 91.9%, X2=0.154, P=0.645). No treatment-related complications were observed in the active surveillance group whereas minor and major complications were observed in 13.0% and 26.1% of the RPLND group, respectively. CONCLUSIONS: Active surveillance resulted in similar DFSR and OSR compared with RPLND in our trial. Patients with low-risk CSI-NSGCTs could benefit from risk-adapted surveillance after these patients were subjected to radical orchidectomy.

[Changes of phenols and lignin contents in alfalfa leaf damaged by Odontothrips loti].

In order to investigate the changes of secondary metabolites content of alfalfa induced by thrips Odontothrips loti damaging, two alfalfa strains, one resistant to thrips (R-1) and the other susceptible to thrips (I-1) , were chosen to measure the phenols and lignin contents of alfalfa leaves under infestation with thrips at different densities (0, 1, 3, 5, 7 thrips x branch(-1), and 0 thrip x branch(-1) as control). After infestation 7 days, the polyphenols, tannin and condensed tannin contents increased in both leaves of R-l and I-i with the increasing thrips density, the simple phenols content had no significant difference, while the lignin content increased significantly compared with the control. After infestation 14 days, the polyphenols, tannin, condensed tannin and lignin contents in both leaves of R-1 and I-1 increased obviously with the increasing thrips density, while the simple phenols content had no significant difference. The lignin content increased significantly, and was significantly higher under 7 thrips x branch(-1) than under the control. After infestation 21 days, the polyphenols, tannin, and lignin contents in both leaves of R-1 and I-1 increased obviously with the increasing thrips density, and were the highest under 7 thrips x branch(-1). Simple phenols content of I-1 strain was increased significantly, but that of R-1 strain had no significant change. The condensed tannin content in both leaves of R-1 and I-1 was not obvious compared with the control. Phenols and lignin contents in R-1 and I-1 leaves increased obviously after thrips infestation, and the polyphenols, tannin and lignin contents increased faster in R-1 strain than in I-1 strain. Thrips infestation had inductive effects on phenols and lignin contents of alfalfa, which could be used to evaluate the resistance of alfalfa.

Neuronal and astroglial TGFß-Smad3 signaling pathways differentially regulate dendrite growth and synaptogenesis.

To address the role of the transforming growth factor beta (TGFß)-Smad3 signaling pathway in dendrite growth and associated synaptogenesis, we used small inhibitory RNA to knockdown the Smad3 gene in either cultured neurons and or primary astrocytes. We found that TGFß1 treatment of primary neurons increased dendrite extensions and the number of synapsin-1-positive synapses. When Smad3 was knockdown in primary neurons, dendrite growth was inhibited and the number of synapsin-1-positive synapses reduced even with TGFß1 treatment. When astrocyte-conditioned medium (ACM), collected from TGFß1-treated astrocytes (TGFß1-stimulated ACM), was added to cultured neurons, dendritic growth was inhibited and the number of synapsin-1-positive puncta reduced. When TGFß1-stimulated ACM was collected from astrocytes with Smad3 knocked down, this conditioned media promoted the growth of dendrites and the number of synapsin-1-positive puncta in cultured neurons. We further found that TGFß1 signaling through Smad3 increased the expression of chondroitin sulfate proteoglycans, neurocan, and phosphacan in ACM. Application of chondroitinase ABC to the TGFß1-stimulated ACM reversed its inhibitory effects on the dendrite growth and the number of synapsin-1-positive puncta. On the other hand, we found that TGFß1 treatment caused a facilitation of Smad3 phosphorylation and translocation to the nucleus induced by status epilepticus (SE) in wild-type (Smad3(+/+)) mice, and this treatment also caused a promotion of γ-aminobutyric acid-ergic synaptogenesis impaired by SE in Smad3(+/+) as well as in Smad3(-/-) mice, but more dramatic promotion in Smad3(+/+) mice. Thus, we provide evidence for the first time that TGFß-Smad3 signaling pathways within neuron and astrocyte differentially regulate dendrite growth and synaptogenesis, and this pathway may be involved in the pathogenesis of some central nervous system diseases, such as epilepsy.

High serum interleukin-6 level is associated with increased risk of delirium in elderly patients after noncardiac surgery: a prospective cohort study.

BACKGROUND: The relationship between inflammation and delirium remains to be determined. The purposes of this study were to investigate the association between serum interleukin-6 levels and the occurrence of delirium in elderly patients after major noncardiac surgery. METHODS: A total of 338 elderly patients (60 years of age and over) undergoing major noncardiac surgery were enrolled. Blood samples were obtained before anesthesia and in the first postoperative morning and serum interleukin-6 concentrations were measured. Delirium was assessed twice daily by the confusion assessment method for the Intensive Care Unit during the first three postoperative days. Survival analyses were performed to assess the relationship between the serum IL-6 level and the occurrence of postoperative delirium. RESULTS: Postoperative delirium occurred in 14.8% (50 of 338) of patients. High serum interleukin-6 levelsafter surgery were significantly associated with increased risk of the occurrence of postoperative delirium (hazard ratio 1.514, 95% confidence interval 1.155-1.985, P = 0.003). Other independent predictors of delirium included increasing age, poor preoperative New York Heart Association classification, low preoperative Mini-Mental State Examination score, and high total postoperative Visual Analogue Scale pain score. Patients who developed delirium had a prolonged hospital stay after surgery. CONCLUSIONS: Delirium is a frequent complication in elderly patients after noncardiac surgery. High serum interleukin-6 level after surgery is associated with increased risk of the occurrence of postoperative delirium.

Solid lipid nanoparticles as delivery systems for Gambogenic acid.

In this study, Gambogenic acid loaded by solid lipid nanoparticles (GNA-SLNs) was explored to reduce toxicity and improve therapeutic efficacy. GNA-SLNs were prepared by emulsification and low temperature solidification methods, and the freeze-dried powders were then developed to improve the stability. The physical-chemical properties of the products in terms of particle size, zeta potential, morphology and entrapment efficiency were well evaluated. The results revealed that the mean diameter, polydispersivity index (PI), zeta potential, and the entrapment efficiency of the nanoparticles were 163.3 nm, 0.203, -16.9 mV and 61.2%, respectively. In comparion with GNA-SLNs, the freeze-dried solid lipid nanoparticles (SLNs) showed a slight augmentation in the mean particle size (from 163.3 to 173 nm) and PI (from 0.203 to 0.253), and no significant modification in the zeta potential, entrapment efficiency and drug loading. In vitro release kinetics based on a dialysis method demonstrated that Gambogenic acid (GNA) was released in a prolonged fashion for 96 h and followed Higuchi equation unitarily. The release profile did not show any significant modification after the freeze-drying process. The Pharmacokinetic study was carried out, the i.p. administration of GNA formulations to rats at doses of 2.5mg/kg. AUC((0-t)) was increased (up to 3.1-fold) and clearance was decreased (up to 3.03-fold) when GNA entrapped in SLNs. In conclusions, the freeze-dried powders form could enhance the long-term stability of SLN, and solid lipid nanoparticles encapsulation could effectively strategy to change the poor aqueous solubility and prolong the half-life of GNA.

Gambogenic acid-induced time- and dose-dependent growth inhibition and apoptosis involving Akt pathway inactivation in U251 glioblastoma cells.

Glioblastoma multiforme is the most common and aggressive type of primary brain tumor. Uncontrolled activation of the PI3K/Akt signaling pathway resulting from genetic alterations in phosphatase and tensin homolog deleted on chromosome 10 (PTEN) and epidermal growth factor receptor (EGFR) correlates with poor prognosis and resistance to chemotherapy and radiotherapy of glioblastomas. In this study, we found that gambogenic acid (GNA), a polyprenylated xanthone isolated from the traditional medicine gamboge, efficiently arrested the cell cycle at the G(0)/G(1) phase by specifically repressing the expression of cyclin D1 and cyclin E, suppressed cell proliferation, colony formation and cell migration, and induced caspase-dependent apoptosis in U251 glioblastoma cells in a time- and dose-dependent manner. The pro-apoptotic effect of GNA on U251 cells was shown to be mediated through inactivation of the Akt pathway, because GNA efficiently suppressed the expression level of EGFR and reduced the phosphorylation of Akt (T308) and GSK3ß (S9). Furthermore, the combined treatment with LY294002, a specific inhibitor of the PI3K/Akt kinase pathway, and GNA showed a synergistic or additive effect on the growth of U251 cells. Our results showed that GNA is a promising therapeutic agent for glioblastomas.

Gambogenic acid inhibits proliferation of A549 cells through apoptosis inducing through up-regulation of the p38 MAPK cascade.

Gamboge is a dry resin secreted from Garcinia hanburryi, and gambogenic acid (GNA) is one of the main active compounds of gamboge. We have previously demonstrated the anticancer activity of GNA in A549 cells and pointed out its potential effects in anticancer therapies. Previous studies reported that GNA induced apoptosis in many cancer cell lines and inhibited A549 tumor growth in xenograft of nude mice in vivo. However, the anticancer mechanism of GNA has still not been well studied. In this paper, we have investigated whether GNA-induced apoptosis is critically mediated by the p38 mitogen-activated protein kinase (MAPK) pathway. Our findings revealed that GNA could induce apoptosis, inhibit proliferation, down-regulate the expression of p38 and MAPK, increase the activations of caspase-9, caspase-3, and cytochrome c release. Furthermore, using SB203580, an adenosine triphosphate-competitive inhibitor of p38 MAPK, inhibit the expression of p-p38 and the experimental results show that it may promote the occurrence of apoptosis induced by GNA. Taken together, these results suggested that up-regulation of the p38 MAPK cascade may account for the activation of GNA-induced apoptosis.

[Amelioration of myocardial ischemia/reperfusion injury by leptin pretreatment and ischemic preconditioning in mouse].

OBJECTIVE: To study the effects and role of leptin pretreatment and ischemic preconditioning (IPC) on the reduction of myocardial ischemia/reperfusion injury (MIRI) in mouse. METHODS: Thirty-six male C57BL/6 mice were randomized into five groups: (1)sham operation group (n=12); (2) brief ischemia/reperfusion (I/R) treatment group (n=6), in which the mice were subjected to three cycles of a 3-minute regional ischemia followed by a 5-minute reperfusion (I/R cycle); (3) MIRI group (n=6), in which MIRI was established in the mice by blocking anterior descending branch of left coronary artery for 30 minutes, followed by 120 minutes reperfusion; (4) IPC group (n=6), in which three I/R cycles were performed on the mice followed by the MIRI protocol; (5) leptin pretreatment group (leptin group, n=6), in which 50 microg/kg of leptin was injected intraperitoneally to the mice 30 minutes before myocardial ischemia. From 6 mice of the sham operation group and from the mice of brief I/R group, serum samples were collected at different time points (0, 5, 30 and 120 minutes) after reperfusion to measure changes in serum leptin level. From the rest of the mice, blood and heart samples were harvested at 120 minutes after reperfusion to analyze the myocardial function and infarct size, leptin, myeloperoxidase (MPO), tumor necrosis factor-alpha (TNF-alpha), and interleukin-6 (IL-6) levels. RESULTS: IPC procedure resulted in an increase in serum leptin level shortly after reperfusion. The leptin level increased significantly than that of sham operation group [(6.24+/-2.34) microg/L vs. (1.35+/-0.45) microg/L] at 5 minutes after I/R, and reached the peak value of (12.36+/-1.33) microg/L at 30 minutes after I/R. Then it gradually decreased to its original (0 minute after reperfusion) value [(1.96+/-1.33) microg/L] at 120 minutes after I/R. There was no difference compared with sham operation group [(1.16+/-0.25) microg/L, P>0.05]. Administration of leptin or IPC before MIRI significantly reduced infarct size [(11.50+/-2.26)%, (9.00+/-1.90)% vs. (37.00+/-2.53)%], the myocardial leptin levels [(8.36+/-3.42) microg/g, (6.71+/-2.03) microg/g vs. (15.51+/-3.92) microg/g], MPO [(17.10+/-3.95) microg/g, (13.33+/-2.88) microg/g vs. (30.83+/-4.06) microg/g], serum leptin levels [(15.03+/-1.87) microg/L, (11.85+/-0.72) microg/L vs. (29.55+/-2.31) microg/L], serum TNF-alpha [(35.10+/-10.12) ng/L, (27.04+/-5.18) ng/L vs. (81.34+/-14.20) ng/L], and IL-6 levels [(167.39+/-72.83) ng/L, (149.13+/-37.69) ng/L vs. (477.30+/-29.09) ng/L, all P<0.01]. CONCLUSION: Pretreatment using leptin results in preconditioning-like tolerance against infarction dysfunction. This cardiac protection effect is mediated, in part, via suppression of inflammation in preconditioned myocardium.

Time trends of co-existing atopic conditions in Singapore school children: prevalence and related factors.

Atopic diseases are known to co-exist and are interrelated. Their co-existence in a patient increases disease burden and morbidity. We sought to study the prevalence of co-existing atopic conditions over time and its associated risk factors among Singapore children. Data was collected from two respective The International Study of Asthma and Allergies in Childhood surveys conducted in 1994 and 2001. Data on asthma, rhinitis and eczema were obtained from the responses to questions about wheezing, exercise-induced wheezing, nocturnal cough, rhinitis symptoms, rash, and diagnoses of asthma, rhinitis and eczema in the last 12 months. Data on demographic and known risk factors were also obtained. The prevalence of children who have had more than one atopic disorder increased significantly from 6.0% in 1994 to 10.2% in 2001 (p < 0.001). Parental history of atopic disease, maternal educational level, male gender and smoke exposure were significantly associated with co-existent atopic symptoms in the child. Among Singapore schoolchildren, there was a significant prevalence of co-existent atopic disease and this prevalence was rising. The influences of both genetic and environmental factors were likely to have contributed to these observations.

[Analysis of xanthones in gamboge by HPLC-PDA-ESI/MS].

The MS and multi-MS spectra of gambogic acid and gambogenic acid in positive ion detection mode were analyzed by electrospray ion trap mass spectrometry (ESI-QITMS) and their cleavage patterns were summarized. Gamboge samples were separated by a Kromasil C18 column and analyzed by HPLC-PDA and MS. Sixteen xanthones could be separated and detected, A collision induced dissociation (CID) experiment was carried out. Molecular weight and UV spectra with of these compounds were obtained. Ten xanthone compounds in Gamboge were identified by online photodiode array detection-MS(n) and by comparing with data from literature. It is expected to develop a comprehensive quality control method for this kind of compounds in commonly used herbal preparation especially in structure analysis of trace substances.