[Effects of salivary microbiota on tryptophan-aryl hydrocarbon receptor signaling axis in mice with periodontitis].

Objective: To study the effects of salivary microbiota in patients with periodontitis on the tryptophan-aryl hydrocarbon receptor (AhR) signaling axis in mice with periodontitis and to provide theoretical basis as well as new ideas for the influences of periodontitis on systemic metabolism. Methods: Salivary microbiota of 12 healthy individuals and 14 patients with periodontitis were collected in Nanjing Stomatological Hospital, Medical School of Nanjing University from June to December of 2020. According to the random number table method, twenty-four mice were randomly divided into three groups: Sham group (control group), P group (periodontitis patients' salivary microbiota group) and H group (periodontal healthy individuals' salivary microbiota group). The maxillary second molars of all mice were treated with silk thread ligation to induce periodontitis. Phosphate buffer as well as salivary microbiota of periodontal healthy individuals and periodontitis patients were gavaged into periodontitis mice for 2 weeks. The expression of inflammatory factors in mice serum were detected by enzyme linked immunosorbent assay, and the expression of tryptophan and indole metabolites in intestinal tract and serum were detected by liquid chromatography-mass spectrometry. The expression of AhR in intestinal tract of mice was detected by immunohistochemistry and quantitative real time-PCR while gut microbiota constitution was detected by 16S rRNA gene sequencing. The remaining saliva samples of periodontitis patients and periodontal healthy individuals were applied to detect the expression of tryptophan and indole metabolites themselves. Results: The salivary microbiota of periodontitis patients could induce the expression of interleukin-1ß [P group: (162.38±39.46) pg/ml, H group: (82.83±20.01) pg/ml; t=4.40, P=0.001) and tumor necrosis factor-α [P group: (361.16±123.90) pg/ml, H group: (191.66±106.87) pg/ml; t=2.54, P=0.030) in serum of periodontitis mice, and reduce the expression of AhR in colon (P group: 1.18±0.05, H group:1.83±0.47; t=3.09, P=0.015) and ileum (P group: 0.80±0.13, H group: 1.18±0.11; t=4.93, P=0.001). After gavage of salivary microbiota of periodontitis patients to the mice, tryptophan (P group: (18.1±3.8)×107, H group: (26.6±6.6)×107; t=2.49, P=0.037] and indole lactic acid [P group: (1.9±0.7)×107, H group: (3.7±0.6)×107; t=4.49, P=0.002) in serum of periodontitis mice were significantly decreased, but was relatively disorder in intestinal tract. However, the expressions of tryptophan and indole metabolites in saliva of periodontitis patients were higher than those of periodontal healthy individuals. There were significant differences in indole propionic acid [P group: (1 239.39±818.72) nmol/L, H group: (56.96±38.33) nmol/L; t=2.83, P=0.022]. What we find noteworthy was that the expressions of indolelactic acid metabolism in saliva, serum and intestinal were consistent, and salivary microbiota of periodontitis patients could reduce the relative abundance of indolelactic acid-producing bacteria in the gut, suggesting that the salivary microbiota of periodontitis patients might affect the expression of AhR through gut microbiota disorder and indolelactic acid downregulation. Conclusions: Salivary microbiota in patients with periodontitis may affect the systemic inflammatory state through down-regulating the expression of tryptophan-AhR signal axis.

Pemetrexed plus platinum with or without pembrolizumab in patients with previously untreated metastatic nonsquamous NSCLC: protocol-specified final analysis from KEYNOTE-189.

BACKGROUND: In the phase III KEYNOTE-189 study (NCT02578680), pembrolizumab plus pemetrexed and platinum-based chemotherapy (pemetrexed-platinum) significantly improved overall survival (OS) and progression-free survival (PFS) in patients with previously untreated metastatic nonsquamous non-small-cell lung cancer (NSCLC) versus placebo plus pemetrexed-platinum. We report updated efficacy outcomes from the protocol-specified final analysis, including outcomes in patients who crossed over to pembrolizumab from pemetrexed-platinum and in patients who completed 35 cycles (∼2 years) of pembrolizumab. PATIENTS AND METHODS: Eligible patients were randomized 2 : 1 to receive pembrolizumab 200 mg (n = 410) or placebo (n = 206) every 3 weeks (for up to 35 cycles, ∼2 years) plus four cycles of pemetrexed (500 mg/m2) and investigators' choice of cisplatin (75 mg/m2) or carboplatin (area under the curve 5 mg·min/ml) every 3 weeks, followed by pemetrexed until progression. Patients assigned to placebo plus pemetrexed-platinum could cross over to pembrolizumab upon progression if eligibility criteria were met. The primary endpoints were OS and PFS. RESULTS: After a median follow-up of 31.0 months, pembrolizumab plus pemetrexed-platinum continued to improve OS [hazard ratio (HR), 0.56; 95% confidence interval (CI), 0.46-0.69] and PFS (HR, 0.49; 95% CI, 0.41-0.59) over placebo plus pemetrexed-platinum regardless of programmed death-ligand 1 expression. Objective response rate (ORR) (48.3% versus 19.9%) and time to second/subsequent tumor progression on next-line treatment (PFS2; HR, 0.50; 95% CI, 0.41-0.61) were improved in patients who received pembrolizumab plus pemetrexed-platinum. Eighty-four patients (40.8%) from the placebo plus pemetrexed-platinum group crossed over to pembrolizumab on-study. Grade 3-5 adverse events occurred in 72.1% of patients receiving pembrolizumab plus pemetrexed-platinum and 66.8% of patients receiving placebo plus pemetrexed-platinum. Fifty-six patients completed 35 cycles (∼2 years) of pembrolizumab; ORR was 85.7% and 53 (94.6%) were alive at data cut-off. CONCLUSIONS: Pembrolizumab plus pemetrexed-platinum continued to show improved efficacy outcomes compared with placebo plus pemetrexed-platinum, with manageable toxicity. These findings support first-line pembrolizumab plus pemetrexed-platinum in patients with previously untreated metastatic nonsquamous NSCLC.

[Current status and research progress of third-line treatment for patients with gastric cancer in China].

Gastric cancer (GC) is one of the most common malignant tumors in the world. The prognosis of advanced GC is extremely poor, characterized by the high recurrence or disease progression rate after the first-line chemotherapy, and the extremely low long-term survival rate. Meanwhile, the options for subsequent treatment are limited. Studies have shown that the third-line therapy can provide significant survival benefits for selected patients with advanced GC. Currently, a series of randomized controlled trials and real-world studies related to chemotherapy, targeted therapy, and immunotherapy are conducted. In addition, the explorations of combination therapy, and screening the optimal clinical features or predictive biomarkers for the suitable population who might benefit from the third-line regimens are the hot spots for researchers. This article will provide a detailed overview of the current status and progress of the third-line treatment for advanced GC, and to illustrate the characteristics of Chinese GC treatment.

Perioperative outcomes of women with and without class III obesity undergoing hysterectomy for endometrioid endometrial cancer: A population-based study.

OBJECTIVE: Population-based data on perioperative complications among women with endometrial cancer and severe obesity are lacking. We evaluated 30-day complication rates among women with and without class III obesity (body mass index ≥ 40 kg/m2) undergoing primary surgical management for endometrioid endometrial cancer (EEC), and how outcomes differed according to surgical approach (open vs. minimally invasive). METHODS: We performed a retrospective population-based cohort study of women with EEC undergoing hysterectomy in Ontario, Canada, between 2006 and 2015. We evaluated perioperative complications in the whole cohort, and in a 1:1 matched analysis using hard and propensity score matching to ensure similar distributions of patient, tumour, provider and institution-level factors between women with and without class III obesity (identified using a surgical billing code). The primary outcome of interest was the 30-day perioperative complication rate. RESULTS: 12,112 women met inclusion criteria; 2697 (22.3%) had class III obesity. We matched 2320 (86%) women with class III obesity to those without. The composite complication rate was significantly higher among women with class III obesity (23.2% vs. 18.4%, standardized mean difference [SMD] = 0.12), primarily due to wound infection/disruption (12.1% vs. 6.2%). There was no difference in outcomes for women with and without class III obesity when a minimally invasive approach was used. CONCLUSIONS: Wound infection/disruption was increased for women with class III obesity compared to women without. Otherwise, perioperative complications were similar between the matched pairs. When minimally invasive approaches were used, women with class III obesity had a similar risk of complications as women without obesity.

Rhamnus prinoides (gesho) stem extract prevents co-culture biofilm formation by Streptococcus mutans and Candida albicans.

Streptococcus mutans and Candida albicans exhibit a symbiotic relationship to form polymicrobial biofilms that exacerbate oral infections including early-childhood caries, periodontitis and candidiasis. Rhamnus prinoides (gesho) has traditionally been used for the treatment of a variety of illnesses and was recently found to inhibit Gram-positive bacterial biofilm formation. We hypothesized that Rhamnus prinoides extracts have anti-biofilm activity against S. mutans and C. albicans mono- and dual-species biofilms. Ethanol extracts were prepared from gesho stems and leaves; then anti-biofilm activity was assessed using crystal violet, resazurin and XTT staining. Ethanol extracts significantly inhibited Streptococcus mutans and Candida albicans mono-species biofilm formation up to 97 and 75%, respectively. The stem ethanol extract disrupted S. mutans and C. albicans co-culture synergism, with 98% less polymicrobial biofilm formation than the untreated control. Additionally, this extract inhibited planktonic S. mutans cell growth and decreased biofilm polysaccharide production up to 99%. The reduction in polysaccharide production is likely a contributing factor in the anti-biofilm activity of GSE. These findings indicate that gesho or gesho-derived compounds may have potential as additives to oral hygiene products. SIGNIFICANCE AND IMPACT OF THE STUDY: Oral Streptococcus mutans and Candida albicans biofilms are associated with a variety of illnesses. When occurring together, the resulting infections are especially challenging to treat due to enhanced biofilm formation and antibiotic resistance. More therapeutics that can effectively prevent polymicrobial biofilm formation and disrupt interspecies synergism are needed. Rhamnus prinoides ethanol extracts significantly inhibited dual-species biofilm formation and disrupted interspecies synergism.

The generation of two specific cancer costing algorithms using Ontario administrative databases.

Cancer treatment and management have become increasingly economically burdensome. Consequently, to help with planning health service delivery, it is vital to understand the associated costs. Administrative databases can be used to help understand and generate real-world system-level costs. Using databases to generate costs can take one of two approaches: top-down or bottom-up. Top-down approaches disaggregate the total health care spending from a global health care budget by sector and provider. A bottom-up approach begins with individual-level health care use and its costs, which are then aggregated.

Temperature and excitation wavelength dependence of circular and linear photogalvanic effect in a three dimensional topological insulator Bi2Se3.

The circular (CPGE) and linear photogalvanic effect (LPGE) of a three-dimensional topological insulator Bi2Se3 thin film of seven quintuple layers excited by near-infrared (1064 nm) and mid-infrared (10.6 [Formula: see text]m) radiations have been investigated. The comparison of the CPGE current measured parallel and perpendicular to the incident plane, together with the comparison of the CPGE current under front and back illuminations, indicates that the CPGE under front illumination of 1064 nm light is dominated by the top surface states of the Bi2Se3 thin film. The CPGE current excited by 10.6 [Formula: see text]m light is about one order larger than that excited by 1064 nm light, which may be attributed to the smaller cancelation effect of the CPGE generated in the two-dimensional electron gas when excited by 10.6 [Formula: see text]m light. Under the excitation of 1064 nm light, the LPGE current is dominated by the component which shows an even parity of incident angles, while the LPGE current excited by 10.6 [Formula: see text]m light is mainly contributed by the component which is an odd parity of incident angles. Both of the CPGE and LPGE currents excited by 1064 nm decrease with increasing temperature, which may be owing to the decrease of the momentum relaxation time and the stronger electron-electron scattering with increasing temperature, respectively.

Effects of an Oral-Pharyngeal Motor Training Programme on Children with Obstructive Sleep Apnea Syndrome in Hong Kong: A Retrospective Pilot Study.

BACKGROUND: This study aimed to investigate the effects of an oropharyngeal motor training programme on children with Obstructive Sleep Apnea Syndrome (OSAS) in Hong Kong. METHODS: In this retrospective study, we reviewed the outcomes of 10 children with OSAS who had received an oropharyngeal motor training programme in Occupational Therapy Department of an acute hospital in Hong Kong over a 1-year programme. Each participant attended an individual oropharyngeal motor training programme plus a follow-up session after 2 months. The training programme consisted of 10 individual mobilization exercises involving the orofacial and pharyngeal area for 45 minutes. Each exercise had to be repeated for 10 times. Three outcome measures were chosen to study the effectiveness of the training programme including tongue strength, tongue endurance level and orofacial function. Tongue strength and tongue endurance level were assessed using the Iowa Oral Pressure Instrument (IOPI). The Nordic Orofacial Test-Screening (NOT-S) Assessment was used to assess the orofacial function. Seven out of 10 participants completed the training programme and attended the follow-up session after two months. RESULTS: The tongue strength and the scores of NOT-S of the 7 participants were found to have significant improvement after training. However, there was no significant difference in tongue endurance level. CONCLUSION: The findings of this study support the role of occupational therapist in oromotor training modalities to improve the respiratory function for children with OSAS in Hong Kong. Copyright © 2017, Hong Kong Occupational Therapy Association. Published by Elsevier (Singapore) Pte Ltd. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

[Second-line treatment for metastatic or locally advanced gastric cancer].

Gastric cancer is one of the major causes of cancer-related deaths. Many patients with metastatic gastric cancer after first-line chemotherapy received salvage chemotherapy in routine clinical practice. Recent phase Ⅲ trials demonstrated substantial prolongation of overall survival to support this chemotherapy or targeted therapy as a second-line treatment. Both ramucirumab monotherapy and ramucirumab plus paclitaxel were approved by FDA in patients with previously treated advanced gastric or gastroesophageal junction adenocarcinoma. In addition, paclitaxel, irinotecan, or docetaxel monotherapy is also recommended for preferred regimens. This review will summarize chemotherapy or targeted therapy as a second-line treatment in advanced gastric cancer.

Radiation costing methods: a systematic review.

OBJECTIVE: Costs for radiation therapy (rt) and the methods used to cost rt are highly diverse across the literature. To date, no study has compared various costing methods in detail. Our objective was to perform a thorough review of the radiation costing literature to identify sources of costs and methods used. METHODS: A systematic review of Ovid medline, Ovid oldmedline, embase, Ovid HealthStar, and EconLit from 2005 to 23 March 2015 used search terms such as "radiation," "radiotherapy," "neoplasm," "cost," " cost analysis," and "cost benefit analysis" to locate relevant articles. Original papers were reviewed for detailed costing methods. Cost sources and methods were extracted for papers investigating rt modalities, including three-dimensional conformal rt (3D-crt), intensity-modulated rt (imrt), stereotactic body rt (sbrt), and brachytherapy (bt). All costs were translated into 2014 U.S. dollars. RESULTS: Most of the studies (91%) reported in the 33 articles retrieved provided rt costs from the health system perspective. The cost of rt ranged from US$2,687.87 to US$111,900.60 per treatment for imrt, followed by US$5,583.28 to US$90,055 for 3D-crt, US$10,544.22 to US$78,667.40 for bt, and US$6,520.58 to US$19,602.68 for sbrt. Cost drivers were professional or personnel costs and the cost of rt treatment. Most studies did not address the cost of rt equipment (85%) and institutional or facility costs (66%). CONCLUSIONS: Costing methods and sources were widely variable across studies, highlighting the need for consistency in the reporting of rt costs. More work to promote comparability and consistency across studies is needed.

[Clinical value of T-cell interferon releases detection of tuberculosis infection assay in quick diagnosis of spinal tuberculosis].

OBJECTIVE: To evaluate the value of T-cell interferon releases detection of tuberculosis infection(T-SPOT.TB)assay in quick diagnosis of spinal tuberculosis. METHODS: From January 2012 to June 2015, a group of 122 diagnosed patients with spinal tuberculosis in the Qingdao Municipal Chest Hospital and a group of 86 patients suspected with spinal tuberculosis in Department of Orthopaedic, the Qingdao Third People's Hospital were accepted to undergone TB-DOT, T-SPOT.TB and TB-DNA PCR tests Department of Clinical Laboratory. RESULTS: The sensitivity of TB-DOT, T-SPOT.TB and TB-DNA PCR tests were 69.7%, 86.1% and 56.6%, respectively.The sensitivity of T-SPOT.TB was significantly higher than TB-DOT and TB-DNA PCR tests (χ(2)=9.51, P<0.05; χ(2)=25.96, P<0.05). The specificity of TB-DOT, T-SPOT.TB and TB-DNA PCR tests were 62.8%, 88.3% and 91.9%, respectively.The specificity of T-SPOT.TB was significantly higher than TB-DOT test (χ(2)=15.25, P<0.05). CONCLUSIONS: T-SPOT.TB assay possesses high sensitivity and specificity in quick diagnosis of patients with spinal tuberculosis, which is valuable in diagnosis of spinal tuberculosis.

SHP-2-upregulated ZEB1 is important for PDGFRα-driven glioma epithelial-mesenchymal transition and invasion in mice and humans.

Gliomas are highly malignant brain tumors that are highly invasive and resistant to conventional therapy. Receptor tyrosine kinases (RTKs) such as PDGFRα (platelet-derived growth factor receptor-α), which show frequent aberrant activation in gliomas, are associated with a process of epithelial-mesenchymal transition (EMT), a cellular alteration that confers a more invasive and drug-resistant phenotype. Although this phenomenon is well documented in human cancers, the processes by which RTKs including PDGFRα mediate EMT are largely unknown. Here, we report that SHP-2 (encoded by PTPN11) upregulates an EMT inducer, ZEB1, to mediate PDGFRα-driven glioma EMT, invasion and growth in glioma cell lines and patient-derived glioma stem cells (GSCs) using cell culture and orthotopic xenograft models. ZEB1 and activated PDGFRα were coexpressed in invasive regions of mouse glioma xenografts and clinical glioma specimens. Glioma patients with high levels of both phospho-PDGFRα (p-PDGFRα) and ZEB1 had significantly shorter overall survival compared with those with low expression of p-PDGFRα and ZEB1. Knockdown of ZEB1 inhibited PDGFA/PDGFRα-stimulated glioma EMT, tumor growth and invasion in glioma cell lines and patient-derived GSCs. PDGFRα mutant deficient of SHP2 binding (PDGFRα-F720) or phosphoinositide 3-kinase (PI3K) binding (PDGFRα-F731/42), knockdown of SHP2 or treatments of pharmacological inhibitor for PDGFRα-signaling effectors attenuated PDGFA/PDGFRα-stimulated ZEB1 expression, cell migration and GSC proliferation. Importantly, SHP-2 acts together with PI3K/AKT to regulate a ZEB1-miR-200 feedback loop in PDGFRα-driven gliomas. Taken together, our findings uncover a new pathway in which ZEB1 functions as a key regulator for PDGFRα-driven glioma EMT, invasiveness and growth, suggesting that ZEB1 is a promising therapeutic target for treating gliomas with high PDGFRα activation.

Expression Patterns of Three Genes Under Short and Long Term Cold Exposure in Thitarodes pui (Lepidoptera: Hepialidae), A Host of Ophiocordyceps sinensis.

　　BACKGROUND: Thitarodes larvae are the host of the caterpillar fungus Ophiocordyceps sinensis. Low temperature is the main environmental limitation for larvae growth. OBJECTIVE: To better understand the cold adaption process in T. pui larvae, the expression patterns of trehalose-6-phosphate synthase (TpTPS), heat shock protein 70 (TpHSP70), and heat shock protein 90 (TpHSP90) were investigated upon short and long-term exposure to 0°C. MATERIALS AND METHODS: The 6th instar T. pui larvae were collected in July 2013. TpTPS was firstly sequenced and expression patterns of TpTPS, TpHSP70 and TpHSP90 were investigated using quantitative PCR. RESULTS: Full-length cDNA of TpTPS was 3,012 bp, with an open reading frame of 2,472 bp and an encoding protein of 823 amino acids. TpTPS up-regulation was induced by cold exposure. TpHSP70 expression is altered by cold exposure, but remained low. TpHSP90 expression was obviously up regulated in long-term cold stimulation. CONCLUSION: All three genes (TpTPS, TpHSP70 and TpHSP90) have likely contributed to cold tolerance in T. pui larvae, TpTPS and TpHSP90 potentially being more important.

Molecular cloning and characterization of GbMECT and GbMECP gene promoters from Ginkgo biloba.

Ginkgolides are key pharmaceutical components in Ginkgo biloba. Using the cDNA sequence of the MECP and MECT genes to design primers, we obtained the promoters of these genes from Ginkgo genomic DNA using the genome walking method. The two promoters were 744 and 982 bp in length, respectively. The cis-elements of the GbMECPs and GbMECT promoters were predicted and analyzed using the plant cis-acting regulatory element database. We found major cis-elements in the sequence of the GbMECT and GbMECPs promoters. The GbMECP promoter contains six TATA boxes and eight CAAT boxes. The GbMECT contains five TATA boxes and seven CAAT boxes. Furthermore, some cis-elements in the promoters of GbMECPs and GbMECT included hormone and light-regulated elements, UB-B-induced elements, and stress-related dehydration-responsive elements. Expression analysis results showed that the MECP gene is mainly involved in responses to CCC (cycocel) and UV-B, and that MECT is mainly involved in responses to wounding treatment. These results also showed that the expression model was consistent with the cis-elements present. During the annual growth cycle, the level of GbMECPs was significantly correlated with terpene lactones accumulation in leaves. A fitted quadratic curve showed the best model for correlating GbMECPs with terpene lactones in leaves. These results will help us to understand the transcriptional regulatory mechanisms involved in key gene expression and ginkgolide accumulation in G. biloba.

Dietary Supplementation of Magnesium Sulfate during Late Gestation and Lactation Affects the Milk Composition and Immunoglobulin Levels in Sows.

This experiment was conducted to investigate the effects of dietary supplementation of magnesium sulfate (MgSO4) during late gestation and lactation on sow and litter performance, fecal moisture, blood biochemistry parameters, immunoglobulin levels and milk composition in sows. Forty-eight sows (Yorkshire×Landrace, 4th to 5th parity) were randomly allocated to 1 of 4 dietary treatments supplemented with 0, 200, 400, or 600 mg/kg MgSO4 (n = 12). The experiment started on day 90 of gestation and continued through day 21 of lactation. Blood samples were collected on day 107 of gestation, day 0 (farrowing) and 21 (weaning) of lactation for the analyses of the blood biochemistry parameters and immunoglobulin levels. The colostrum and milk samples were obtained on day 0 and 14 of lactation, respectively. Fecal samples were collected from the sows on day 107 of gestation as well as day 7 and 20 of lactation to determine fecal moisture content. The results showed that the survival percentage of piglets and the litter weight at weaning were decreased linearly (p<0.05) and other parameters of the sow or litter performance were not influenced (p>0.05) by MgSO4 supplementation. The fecal moisture content of the sows were increased (p<0.05) linearly as dietary MgSO4 increased on day 7 and 20 of lactation. Supplementation with MgSO4 increased the plasma magnesium (Mg) level linearly (p<0.05) and had a trend to increase total protein level (p>0.05 and p<0.10). However, an increase in the dietary MgSO4 level resulted in a linear decrease in the colostrum fat content (p<0.05). Dietary MgSO4 supplementation enhanced the immunoglobulin G (IgG) level (linear, p<0.05) in plasma on day of farrowing and immunoglobulin A (IgA) level in colostrum (quadratic, p<0.05) and milk (linear, p<0.05) of the sows. These results indicated that supplementation with MgSO4 during late gestation and lactation may have the potential to prevent sow constipation, but may also result in some negative effects.

Functional characterization of the Ginkgo biloba chalcone synthase gene promoter in transgenic tobacco.

The regulative sequence (2273 bp) of the chalcone synthase gene promoter of biloba was cloned by genomic walking. A 2273-bp promoter 5' upstream translation start site of GbCHS was cloned and designated as GbCHSP. pBI121+CHSP:GUS and pBI121-35S:GUS were constructed and transformed into tobacco by LBA4404. We found that GbCHSP could drive transient expression of GUS in tobacco and differentially expressed in root, stem and leaf tissues of this plant. GUS activity regulated by the CHSP promoter were located in tissues (apical meristems) at the growing points of roots and stems. pBI121+CHSP:GUS could be induced by wounding, copper, UV-B, abscisic acid, and ethephon treatments of transgenic seedlings. This activity was weakly inhibited by gibberellin. Deletion analysis of the CHSP promoter in transgenic tobacco showed that CHSP1 complete promoter conferred a GUS expression and activity similar to that of 35 S(CaMV). GUS activity dropped dramatically when there were CHSP4, CHSP5 constructs and was almost totally absent when the CHSP6 construct was present. We conclude that the upstream sequence -1548 to -306 of GbCHSP is the main region for transcriptional regulation of the CHS gene and that it is activated by hormone and stress factors in G. biloba. These results will help us to understand the transcriptional regulatory mechanisms involved in GbCHS expression and flavonoid accumulation in G. biloba.

EGFRvIII stimulates glioma growth and invasion through PKA-dependent serine phosphorylation of Dock180.

Glioblastomas (GBMs), the most common and malignant brain tumors, are highly resistant to current therapies. The failure of targeted therapies against aberrantly activated oncogenic signaling, such as that of the EGFR-PI3K/Akt pathway, underscores the urgent need to understand alternative downstream pathways and to identify new molecular targets for the development of more effective treatments for gliomas. Here, we report that EGFRvIII (ΔEGFR/de2-7EGFR), a constitutively active EGFR mutant that is frequently co-overexpressed with EGFR in clinical GBM tumors, promotes glioma growth and invasion through protein kinase A (PKA)-dependent phosphorylation of Dock180, a bipartite guanine nucleotide exchange factor (GEF) for Rac1. We demonstrate that EGFRvIII induces serine phosphorylation of Dock180, stimulates Rac1 activation and glioma cell migration. Treatments of glioma cells using the PKA inhibitors H-89 and KT5720, overexpression of a PKA inhibitor (PKI), and in vitro PKA kinase assays show that EGFRvIII induction of serine phosphorylation of Dock180 is PKA-dependent. Significantly, PKA induces phosphorylation of Dock180 at amino acid residue S1250 that resides within its Rac1-activating DHR-2 domain. Expression of the Dock180(S1250L) mutant, but not wild type Dock180(WT), protein in EGFRvIII-expressing glioma cells inhibited receptor-stimulated cell proliferation, survival, migration in vitro and glioma tumor growth and invasion in vivo. Together, our findings describe a novel mechanism by which EGFRvIII drives glioma tumorigenesis and invasion through PKA-dependent phosphorylation of Dock180, thereby suggesting that targeting EGFRvIII-PKA-Dock180-Rac1 signaling axis could provide a novel pathway to develop potential therapeutic strategies for malignant gliomas.

Effectiveness of maifanite in reducing the detrimental effects of cadmium on growth performance, cadmium residue, hematological parameters, serum biochemistry, and the activities of antioxidant enzymes in pigs.

This study was conducted to investigate the toxicity of cadmium and to evaluate the effectiveness of maifanite in preventing cadmium-induced adverse effects. Thirty-two crossbred pigs (Duroc × Landrace × Large white, sex balanced, 17.25 ± 0.07 kg average body weight) were randomly allotted to one of four dietary treatments in a 2 × 2 factorial arrangement, with eight replicates per treatment and one pig per replicate. The dietary treatments included two cadmium (as CdCl2) doses (0.32 and 30.49 mg/kg) and two maifanite doses (0 and 1%). The results showed that pigs treated with cadmium decreased their average daily feed intake (P < 0.05) and increased (P < 0.05) the feed/gain ratio. Cadmium was found in the tissues of pigs that were fed with cadmium-contaminated diets, but the level of cadmium was much lower when maifanite was added to the cadmium-contaminated diets. Ingestion of diets that were artificially contaminated with cadmium (30.49 mg/kg of cadmium) reduced (P < 0.05) the number of lymphocytes, the total erythrocyte count, the hemoglobin level, and the hematocrit. However, the activities of serum aspartate aminotransferase and gamma glutamyltransferase were increased (P < 0.05). The total protein level was lower (P < 0.05) in pigs fed with cadmium-contaminated diets. The contents of malondialdehyde increased (P < 0.05), while the total antioxidant capacity and the activities of total superoxide dismutase, glutathione peroxidase, glutathione S-transferase, and catalase decreased (P < 0.05) in pigs fed with cadmium-contaminated diets. Dietary addition of maifanite can, to some extent, prevent the negative effects associated with feeding cadmium diets (30.49 mg/kg of cadmium) to pigs.