Risk of thromboembolism in non-small-cell lung cancers patients with different oncogenic drivers, including ROS1, ALK, and EGFR mutations.

BACKGROUND: Anaplastic lymphoma kinase-positive (ALK+) and ROS proto-oncogene 1 (ROS1)-positive (ROS1+) lung cancers have been reported to be associated with an elevated risk of thromboembolic events. This study aimed to assess the long-term risk of developing thromboembolism (TE) in ROS1+ lung cancer and to compare it with other oncogenic drivers in the Asian population. MATERIALS AND METHODS: We retrospectively enrolled a cohort of ROS1+ lung adenocarcinoma in a medical center in Taiwan and a comparison cohort of ALK+ and epidermal growth factor receptor-positive (EGFR+) lung cancers. Venous and arterial TEs were identified throughout the cancer course, and the incidence rate was calculated. RESULTS: We enrolled 44 ROS1+, 98 ALK+, and 168 EGFR+ non-small-cell lung cancer (NSCLC) patients. A total of 11 (25%), 36 (36.7%), and 38 (22.6%) patients in the ROS1, ALK, and EGFR cohorts, respectively, were diagnosed with thromboembolic events throughout the follow-up course of the disease (P = 0.042). The incidence rates were 99.0, 91.9, and 82.5 events per 1000 person-years for the ROS1, ALK, and EGFR cohorts, respectively. The majority of thrombosis events in the ROS1 (91.6%) and ALK (85.4%) cohorts were venous. On the contrary, 43.2% of thromboembolic events were arterial in the EGFR cohort. A higher proportion of thromboembolic events were noted during cancer diagnosis in the ROS1 cohort (36.3%) than in the ALK (16.7%) and EGFR (10.5%) cohorts. The stage was the only clinical variable associated with thromboembolic risk. There was a significant difference in survival between patients with and without TE in the EGFR cohort, but not in the ALK and ROS1 cohorts. CONCLUSIONS: Although ROS1+ and ALK+ NSCLCs had a higher cumulative incidence of TE than EGFR+ NSCLC, the person-year incidence rates were similar among the three groups. EGFR-mutated NSCLC had more arterial events. Nevertheless, ALK+ lung cancer had higher venous events than EGFR-mutated lung cancer.

Recurrence of pericardial effusion after different procedure modalities in patients with non-small-cell lung cancer.

BACKGROUND: Lung cancer with related pericardial effusion is not rare. Intervention is a crucial step for symptomatic effusion. It is unknown, however, whether the different invasive interventions for pericardial effusion result in different survival outcomes. This study analyzed the clinical characteristics and prognostic factors for patients with non-small-cell lung cancer (NSCLC) who have undergone different procedures. METHODS: From January 2006 to June 2018, we collected data from patients with NSCLC who have received invasive intervention for pericardial effusions. The patients were divided into three categories: simple pericardiocentesis, balloon pericardiotomy, and surgical pericardiectomy. Kaplan-Meier curve and log-rank test were used to analyze the pericardial effusion recurrence-free survival (RFS) and overall survival (OS). RESULTS: A total of 244 patients were enrolled. Adenocarcinoma (83.6%) was the major NSCLC subtype. Invasive intervention, including simple pericardiocentesis, balloon pericardiotomy, and surgical pericardiectomy, had been carried out on 52, 170, and 22 patients, respectively. The 1-year RFS rates in simple pericardiocentesis, balloon pericardiotomy, and surgical pericardiectomy were 19.2%, 31.2%, and 31.8%, respectively (P = 0.128), and the median RFS was 1.67, 5.03, and 8.32 months, respectively (P = 0.008). There was no significant difference in OS, however, with the median OS at 1.67, 6.43, and 8.32 months, respectively (P = 0.064). According to the multivariable analysis, the gravity in pericardial fluid analysis, receiving systemic therapy after pericardial effusion, and the time period from stage IV lung cancer to the presence of pericardial effusion were independent prognostic factors for pericardial effusion RFS and OS. CONCLUSIONS: Patients who have undergone simple pericardiocentesis alone for the management of NSCLC-related pericardial effusion have lower 1-year RFS rates than those who have undergone balloon pericardiotomy and surgical pericardiectomy. Therefore, balloon pericardiotomy and surgical pericardiectomy should be carried out for patients with NSCLC-related pericardial effusion if tolerable.

[Clinicopathological characteristics and prognosis of diffuse midline gliomas with histone H3K27M mutation: an analysis of 30 cases].

Objective: To analyze the clinicopathological characteristics and prognosis of diffuse midline glioma (DMG) with H3K27M mutation. Methods: Thirty cases of DMG were collected in Guangdong Sanjiu Brain Hospital from October 2016 to May 2018. The patients' clinicopathological data including age, tumor site and histological grade, treatment and follow-up data were collected and analyzed. Results: There were 21 males and 9 females, with a mean age of 26 years (range 5-53 years). Fourteen tumors were located in thalamus, 12 in brainstem (one involved both thalamus and brainstem), and one each in hypothalamus, fourth ventricle, and sellar region, respectively. Two cases presented as diffuse intracranial lesions. Three cases (10.0%) were of WHO grade Ⅰ, 10 cases (33.3%) were grade Ⅱ, eight cases (26.7%) were grade Ⅲ, and nine cases (30.0%) were grade Ⅳ.All patients with gradeⅠ tumors were older than 20 years. Histologically, all were pilocytic astrocytoma-like. Immunohistochemical staining demonstrated that all tumors were IDH1 negative. Twenty-eight tumors showed diffuse expression of H3K27M, and two showed focal expression. Twenty-one tumors(100.0%, 21/21) showed absent expression of H3K27me3. Sixteen tumors (57.1%, 16/28) showed strongly positive expression of p53, and ATRX was negative in eight tumors (38.1%, 8/21). The Ki-67 proliferation index ranged from 5% to 40%. Eight cases (including two cases of H3K27M expression of individual cells) showed K27M mutation in H3F3A gene. Intracranial and spinal cord dissemination occurred in six cases (20.0%, 6/30). Median progression-free survival (PFS) was 9.5 months and median overall survival (OS) was 34 months. Mean PFS was 11.2 months and mean OS was 24.3 months. Compared with adults (>20 years old), children/adolescents (no more than 20 years old) had significantly shorter median OS (8 months vs. 34 months, P=0.013). There was no significant difference in PFS and OS between DMGs located in the brain stem/thalamus and other sites within midline (P>0.05). There was no significant difference in PFS and OS between WHO grade ⅠDMGs and WHO grade Ⅱ-Ⅳ DMGs (P>0.05). Conclusions: DMGs occur more commonly in children and adolescents with male predominance. DMGs present with WHO Ⅰ-Ⅳ tumors morphologically, and pilocytic astrocytoma-like lesions with WHO Ⅰ are more common in adults. Expression of H3K27M but not H3K27me3 is helpful for diagnosis of DMG. The prognosis of children/adolescents is significantly worse than that of adults, whereas histological grade and tumor location do not affect prognosis.

[Gas chromatography-mass spectrometry based urinary metabolomics in very low birth weight premature infants].

Objective: To investigate the urinary metabolic spectrum and pathways in very low birth weight (VLBW) premature infants. Method: A prospective case-control study was conducted to collect and compare the data of VLBW premature infants and full term infants from the Sixth Affiliated Hospital of Sun Yet-Sen University in 2014. Within 24 hours after birth, urine specimens in each group were collected. Metabolites of urine samples including amino acid, fatty acid and organic acid were detected using the urease pre-processing and gas chromatography mass spectrometry (GC-MS) technology. Using the orthogonal partial least squares discriminant analysis (OPLS-DA), the biomarkers and differences between the two groups were found. The online metabolic pathway website was explored and multivariable analysis was conducted to investigate the valuable pathways and biomarkers related to the prematurity. Result: A total of 20 VLBW premature infants were enrolled, among whom 11 were male, 9 were female; and 20 full term infants were enrolled, among whom 9 were male, 11 were female. The urinary metabolites were established and compared between the VLBW premature and term infants. The investigation showed that the following nine pathways were enriched: amino-acyl-tRNA biosynthesis(P=0.000), lysine degradation(P=0.007), fatty acid biosynthesis(P=0.008), pyrimidine metabolism(P=0.014), pantothenate and CoA biosynthesis(P=0.022), valine, leucine and isoleucine biosynthesis(P=0.022), lysine biosynthesis(P=0.031), glycerolipid metabolism(P=0.046), and valine, leucine and isoleucine degradation(P=0.031). Almost all the metabolites decreased except for the glyceric acid exhibiting a higher content in the VLBW premature infant. 12 potential biomarkers were explored with the most significant covariance and correlation, within which stearic acid, palmiticacid, myristic acid, ß-amino-isobutyric acid, and uric acid were lower, while myo-inositol, mannitol, glycine, glucose1, glucose2, glyceric acid and N-acetyl-tyrosine were higher in the VLBW premature group compared with the control group. Conclusion: There is a significant difference between the VLBW premature infants and full-term infants in the metabolic state and pathways. The urease pre-processing and GC-MS technology followed by the OPLS-DA and multivariable analysis to investigate VLBW premature infants' urinary metabolites is a valuable method to evaluate the patients' metabolism.

Effects of dietary gallic acid supplementation on performance, antioxidant status, and jejunum intestinal morphology in broiler chicks.

Gallic acid (GA), widely distributed in plants and feeds, is known to have a diverse range of activities such as anti-oxidant, anti-inflammatory, antibacterial, anti-allergic, anti-mutagenic, and anti-carcinogenic. The purpose of this study was to investigate the efficacy of inclusion of dietary GA at levels 0, 25, 50, 75, 100, or 150 mg/kg on growth performance, antioxidant status, and jejunum intestinal morphology of broiler chicks. In total, 630 one-day-old Arbor Acres (AA) male broiler chicks were randomly allotted to 6 treatment groups for a period of 6 weeks. The results indicate that dietary addition of GA at 75 to 100 mg/kg improved feed conversion efficiency in both the grower (d 21 to 42, P = 0.045) and overall (d 1 to 42, P = 0.026) periods. Dietary addition of GA at a concentration ≥100 mg/kg was able to exhibit higher breast muscle ratio at 42 d (P = 0.043). Interestingly, dietary GA inclusion level from 50 to 100 mg/kg reduced the crypt depth (P = 0.009) and increased the villus height:crypt depth ratio (VCR) of the birds (P = 0.006). Dietary supplementation of GA at 100 mg/kg decreased plasma malondialdehyde (MDA) content at 42 d of age (P = 0.030). Moreover, dietary addition of GA linearly increased plasma total antioxidant capacity (P = 0.039) and plasma total superoxide dismutase activities (P = 0.049) at 21 d of age. However, analysis of plasma biochemical markers revealed that dietary supplementation of GA did not exhibit beneficial health effects. Overall, we conclude that 75 to 100 mg/kg of GA are suitable for enhanced growth performance and health benefits in a broiler diet.

Benign mixed tumor of the vagina.

Mixed tumor of the vagina is a rare benign tumor containing both epithelial and mesenchymal components. The authors report the case of a 35-year-old woman who presented with a painless mass that had descended from the vagina in the last six months. Physical examination showed the presence of a solitary, non-tender nodule that was 3x3x2 cm in size and seemed to originate from the posterior wall of the lower vagina. The tumor was removed and pathologically examined. The findings were indicative of a mixed tumor of the vagina, and were in agreement with previous cases. No recurrence or progression occurred during the ten-month follow-up. As this tumor shares common features with some other tumors, its differential diagnosis is very important. Moreover, due to the rarity of this disease, gynecologists and pathologists need to familiarize themselves with the features in order to avoid a misdiagnosis.

Effects of dietary octacosanol supplementation on laying performance, egg quality, serum hormone levels, and expression of genes related to the reproductive axis in laying hens.

This experiment was conducted to evaluate the effects of dietary octacosanol supplementation on laying performance, egg quality, serum hormone levels, and gene expression related to reproductive axis in laying hens to confirm the reproduction-promoting function of octacosanol. In total, 360 Hy-Line Brown (67-wk-old) laying hens were randomly assigned to one of three treatments with 0, 5, and 10 mg octacosanol (extracted from rice bran, purity >92%)/kg feed. The feeding trial lasted for 10 weeks. The results showed that the dietary addition of 5 and 10 mg/kg octacosanol improved feed efficiency by 4.9% and 3.4% (P < 0.01), increased the albumen height by 20.5% and 13.3% (P < 0.01), the Haugh unit score by 12.9% and 8.7% (P < 0.01), and the eggshell strength by 39.5% and 24.5% (P < 0.01), respectively, compared with the control diet. Dietary octacosanol addition significantly affected serum triiodothyronine, estradiol, follicle-stimulating hormone levels (P < 0.05), and progesterone and luteinizing hormone level (P < 0.01). Compared with the control, dietary addition of octacosanol at 5 mg/kg promoted the follicle-stimulating hormone receptor (FSHR) mRNA expression in different-sized follicles, and significantly increased the FSHR mRNA expression of granulosa cells from the F2 and F3 follicles (P < 0.05). Dietary supplementation with both 5 and 10 mg/kg octacosanol promoted the mRNA expression of luteinizing hormone receptor and prolactin receptor in different-sized follicles, and significantly up-regulated the expression levels in F1 granulosa cells (P < 0.05). The ovarian weight was significantly increased with the dietary addition of 5 mg/kg octacosanol (P < 0.05). The numbers of small yellow follicles and large white follicles were increased with the addition of dietary 5 and 10 mg/kg octacosanol (P < 0.01). This study provides evidence that octacosanol has the capacity to improve reproductive performance, indicating that it is a potentially effective feed additive in egg production.

Effects of dietary grape proanthocyanidins on the growth performance, jejunum morphology and plasma biochemical indices of broiler chicks.

Grape proanthocyanidins (GPCs) are a family of naturally derived polyphenols that have aroused interest in the poultry industry due to their versatile role in animal health. This study was conducted to investigate the potential benefits and appropriate dosages of GPCs on growth performance, jejunum morphology, plasma antioxidant capacity and the biochemical indices of broiler chicks. A total of 280 newly hatched male Cobb 500 broiler chicks were randomly allocated into four treatments of seven replicates each, and were fed a wheat-soybean meal-type diet with or without (control group), 7.5, 15 or 30 mg/kg of GPCs. Results show that dietary GPCs decrease the feed conversion ratio and average daily gain from day 21 to day 42, increase breast muscle yield by day 42 and improve jejunum morphology between day 21 and day 42. Chicks fed 7.5 and 15 mg/kg of GPCs show increased breast muscle yield and exhibit improved jejunum morphologies than birds in the control group. Dietary GPCs fed at a level of 15 mg/kg markedly increased total superoxide dismutase (T-SOD) activity between day 21 and day 42, whereas a supplement of GPCs at 7.5 mg/kg significantly increased T-SOD activity and decreased lipid peroxidation malondialdehyde content by day 42. A supplement of 30 mg/kg of GPCs has no effect on antioxidant status but adversely affects the blood biochemical indices, as evidenced by increased creatinine content, increased alkaline phosphatase by day 21 and increased alanine aminotransferase by day 42 in plasma. GPC levels caused quadratic effect on growth, jejunum morphology and plasma antioxidant capacity. The predicted optimal GPC levels for best plasma antioxidant capacity at 42 days was 13 to 15 mg/kg, for best feed efficiency during grower phase was 16 mg/kg, for best jejunum morphology at 42 days was 17 mg/kg. In conclusion, GPCs (fed at a level of 13 to 17 mg/kg) have the potential to be a promising feed additive for broiler chicks.

miR-508-5p acts as an anti-oncogene by targeting MESDC1 in hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is the third leading cause of cancer associated mortality. Accumulating evidence has shown that microRNAs (miRNAs) act as critical factors for tumor recurrence and metastasis. MiR-508-5p has been reported as a down-regulated miRNA in the primary gastric cancer tissues. However, the role of miR-508-5p on HCC has not been well elucidated. In this study, we observed that miR-508-5p was downregulated in HCC tissues when compared to the non-tumorous tissues. We then demonstrated that overexpression of miR-508-5p attenuated HepG2 cells proliferation and invasion and induced cell apoptosis in vitro. Furthermore, our further investigations revealed that mesoderm development candidate 1 (MESDC1) is a potential target of miR-508-5p, as well as miR-508-5p overexpression downregulated MESDC1 expression. Overexpression of MESDC1 promoted HepG2 cells migration, invasion and proliferation in vitro. In addition, miR-508-5p markedly suppressed the tumor growth in xenograft model, while MESDC1 promoted the tumor growth in xenograft model. This study provides new insight into molecular mechanisms that miR-508-5p acts as a tumor suppressor by targeting MESDC1 in HCC progression.

Dietary supplementation of pyrroloquinoline quinone disodium protects against oxidative stress and liver damage in laying hens fed an oxidized sunflower oil-added diet.

The protective effects of dietary pyrroloquinoline quinone disodium (PQQ.Na2) supplementation against oxidized sunflower oil-induced oxidative stress and liver injury in laying hens were examined. Three hundred and sixty 53-week-old Hy-Line Gray laying hens were randomly allocated into one of the five dietary treatments. The treatments included: (1) a diet containing 2% fresh sunflower oil; (2) a diet containing 2% thermally oxidized sunflower oil; (3) an oxidized sunflower oil diet with 100 mg/kg of added vitamin E; (4) an oxidized sunflower oil diet with 0.08 mg/kg of PQQ.Na2; and (5) an oxidized sunflower oil diet with 0.12 mg/kg of PQQ.Na2. Birds fed the oxidized sunflower oil diet showed a lower feed intake compared to birds fed the fresh oil diet or oxidized oil diet supplemented with vitamin E (P=0.009). Exposure to oxidized sunflower oil increased plasma malondialdehyde (P<0.001), hepatic reactive oxygen species (P<0.05) and carbonyl group levels (P<0.001), but decreased plasma glutathione levels (P=0.006) in laying hens. These unfavorable changes induced by the oxidized sunflower oil diet were modulated by dietary vitamin E or PQQ.Na2 supplementation to levels comparable to the fresh oil group. Dietary supplementation with PQQ.Na2 or vitamin E increased the activities of total superoxide dismutase and glutathione peroxidase in plasma and the liver, when compared with the oxidized sunflower oil group (P<0.05). PQQ.Na2 or vitamin E diminished the oxidized sunflower oil diet induced elevation of liver weight (P=0.026), liver to BW ratio (P=0.001) and plasma activities of alanine aminotransferase (P=0.001) and aspartate aminotransferase (P<0.001) and maintained these indices at the similar levels to the fresh oil diet. Furthermore, oxidized sunflower oil increased hepatic DNA tail length (P<0.05) and tail moment (P<0.05) compared with the fresh oil group. Dietary supplementation of PQQ.Na2 or vitamin E decreased the oxidized oil diet induced DNA tail length and tail moment to the basal levels in fresh oil diet. These results indicate that PQQ.Na2 is a potential antioxidant and is as effective against oxidized oil-related liver injury in laying hens as vitamin E. The protective effects of PQQ.Na2 against liver damage induced by oxidized oil may be partially due to its role in the scavenging of free radicals, inhibiting of lipid peroxidation and enhancing of antioxidant defense systems.

Treatment outcome of patients with isoniazid mono-resistant tuberculosis.

Isoniazid mono-resistance is the most common first-line drug resistance in tuberculosis (TB), but its treatment outcome remains unclear. From January 2004 to October 2011, 425 (5.1%) of 8414 patients with culture-confirmed pulmonary TB from four hospitals in Taiwan were identified as having isoniazid mono-resistant TB. Among them, 395 (92.9%) were included and followed up for 2 years after complete treatment. Although 328 (83.0%) patients were successfully treated, 67 (17.0%) had unfavourable outcomes, including death in 56 (14.2%) and treatment failure in 11 (2.8%). The treatment success rate was similar in patients with high-level and low-level isoniazid-resistant TB (82.2% versus 83.4%, p 0.785) and among those taking anti-TB treatment with and without isoniazid (83.1% versus 83.0%, p 1.000). Patients without rifampicin interruption had lower risk of unfavourable outcome (14.3% versus 37.0%, p <0.001), especially those with low-level isoniazid resistance (11.5% versus 56.5%, p <0.001). Supplementation with a new-generation fluoroquinolone improved treatment success (60.0% versus 12.5%, p 0.003). The presence of cavitary lesions was significantly associated with a higher relapse rate (4.1% versus 0.0%, p 0.006) and extended treatment of 7-9, 10-12 and >12 months had less relapse than 6-month treatment (3.2%, 0%, 3.7% and 25.0%, respectively, p 0.037). Multivariate Cox proportional hazards analysis revealed that co-morbidity with cancer (hazard ratio, 2.43) and rifampicin interruption (hazard ratio 1.91) were independent factors associated with unfavourable outcomes. Treatment throughout with rifampicin and extended treatment for cavitary disease are crucial for improving outcomes in patients with isoniazid mono-resistant TB.

Effects of dietary pyrroloquinoline quinone disodium on growth performance, carcass yield and antioxidant status of broiler chicks.

Pyrroloquinoline quinone (PQQ), a putative essential nutrient and redox modulator in microorganisms, cell and animal models, has been recognized as a growth promoter in rodents. Growth performance, carcass yield and antioxidant status were evaluated on broiler chickens fed different levels of PQQ disodium (PQQ.Na2). A total of 784 day-old male Arbor Acres (AA) broilers were randomly allotted into seven dietary groups: negative control group (NC) fed a basal diet without virginiamycin (VIR) or PQQ.Na2; a positive control group (PC) fed a diet with 15 mg of VIR/kg diet; and PQQ.Na2 groups fed with 0.05, 0.10, 0.20, 0.40 or 0.80 mg PQQ.Na2/kg diet. Each treatment contained eight replicates with 14 birds each. The feeding trial lasted for 6 weeks. The results showed that chicks fed 0.2 mg PQQ.Na2/kg diet significantly improved growth performance comparable to those in PC group, and the feed efficiency enhancement effects of dietary PQQ.Na2 was more apparent in grower phase. Dietary addition of PQQ.Na2 had the potential to stimulate immune organs development, and low level dietary addition (<0.1 mg/kg) increased plasma lysozyme level. Broilers fed 0.2 mg PQQ.Na2/kg diet gained more carcasses at day 42, and had lower lipid peroxide malondialdehyde content and higher total antioxidant power in plasma. The results indicated that dietary PQQ.Na2 (0.2 mg/kg diet) had the potential to act as a growth promoter comparable to antibiotic in broiler chicks.

Relapse of hyperthyroidism after hemithyroidectomy in concurrent medullary thyroid cancer and Graves' disease.

We present a rare case of concurrent medullary thyroid cancer (MTC) and relapse of Graves' disease (GD). A 26-year-old Chinese female suffered from GD for 1 year. Physical examination demonstrated mildly diffuse goiter with a palpable 1.0 cm nodule in the right thyroid lobe and exophthalmos. Before hemithyroidectomy, calcitonin was elevated and thyroglobulin was normal. After surgery, histopathological findings proved coexistence of MTC and GD. 16 months later, GD relapsed and radioactive iodine 131 therapy was given. Meanwhile, postoperative monitoring showed well controlled calcitonin and carcinoembryonic antigen. Our lesson demonstrates although concomitant of MTC and GD is very rare, yet after hemithyroidectomy for the single nodular MTC lesion, remnant thyroid lobe could be the source of GD relapse very soon. More importantly, the relapse of GD and the following radioiodine therapy and radiation exposure could have been prevented if a better therapy plan of total thyroidectomy was adopted in the first place, instead of hemithyroidectomy (Tab. 1, Fig. 2, Ref. 11).

A comparison of quality of life and satisfaction of women with early-stage breast cancer treated with breast conserving therapy vs. mastectomy in southern China.

BACKGROUND: The purpose of this study was to compare the quality of life (QoL) and overall satisfaction with treatment of women with stage T1-2N0M0 breast cancer treated with breast conserving therapy (BCT) or mastectomy (MAS) in southern China. METHODS: Functional assessment of cancer therapy-breast, traditional Chinese version 4 (FACT-B), was administered to 180 patients with stage T1-2N0M0 breast cancer (82 BCT and 98 MAS) treated between July 2000 and July 2008. RESULTS: The two groups differed in tumor pathology and how axillary lymph nodes were treated (sentinel node biopsy vs. dissection), while other disease and socioeconomic characteristics were similar. The median follow-up after completion of radiotherapy was 60 months in the BCT group, and 65 months in the MAS group. The scores of the physical, functional, and emotional domains and breast-specific concerns of FACT-B were not significantly different between the groups. The social domain score of the BCT group was significantly greater than those of the MAS group. CONCLUSIONS: Patients who underwent BCT did not report better QoL than those who received MAS, but BCT patients experienced easier social adjustment.

RNA is favourable for analysing EGFR mutations in malignant pleural effusion of lung cancer.

Malignant pleural effusion (MPE) is a useful specimen allowing for the evaluation of EGFR status in nonsmall cell lung cancer (NSCLC). However, direct sequencing of genomic DNA from MPE samples was found not to be sensitive for EGFR mutation detection. To test whether EGFR analysis from RNA is less prone to interference from nontumour cells that have no or lower EGFR expression, we compared three methods (sequencing from cell-derived RNA versus sequencing and mass-spectrometric analysis from genomic DNA), in parallel, for EGFR mutation detection from MPE samples in 150 lung adenocarcinoma patients receiving first-line tyrosine kinase inhibitors (TKIs). Among these MPE samples, EGFR mutations were much more frequently identified by sequencing using RNA than by sequencing and mass-spectrometric analysis from genomic DNA (for all mutations, 67.3 versus 44.7 and 46.7%; for L858R or exon 19 deletions, 61.3 versus 41.3 and 46.7%, respectively). The better mutation detection yield of sequencing from RNA was coupled with the superior prediction of clinical efficacy of first-line TKIs. In patients with acquired resistance, EGFR sequencing from RNA provided satisfactory detection of T790M (54.2%). These results demonstrated that EGFR sequencing using RNA as template greatly improves sensitivity for EGFR mutation detection from samples of MPE, highlighting RNA as the favourable source for analysing EGFR mutations from heterogeneous MPE specimens in NSCLC.

Comparison of lipid oxidation, messenger ribonucleic acid levels of avian uncoupling protein, avian adenine nucleotide translocator, and avian peroxisome proliferator-activated receptor-γ coactivator-1α in skeletal muscles from electrical- and gas-stunned broilers.

The aim of this study was to compare the effects of stunning methods [electrical stunning (ES) vs. gas stunning (GS)] on lipid oxidation in broiler meat and to investigate possible mechanisms of lipid oxidation by measuring plasma variables, muscle reactive oxygen species (ROS), and TBA reactive substance (TBARS) concentrations, muscle fiber ratios, and mRNA levels of avian uncoupling protein (avUCP), avian adenine nucleotide translocator, and avian peroxisome proliferator-activated receptor-γ coactivator-1α (avPGC-1α). Arbor Acres broilers (n = 36) were not stunned (control) or were exposed to the following stunning treatments: 40% CO(2) + 21% O(2) + N(2); 60% CO(2) + 21% O(2) + N(2); 35 V, 47 mA, 400 Hz; 50 V, 67 mA, 160 Hz; and 65 V, 86 mA, 1,000 Hz. The ROS level in tibialis anterior (TA; P < 0.05) and the TBARS concentration in pectoralis major (PM; P < 0.01) were decreased in the GS groups compared with the ES groups at 45 min postmortem. However, the TBARS concentrations at 24 h postmortem in the PM and TA groups were not affected by stunning method (ES or GS). Compared with ES, GS caused greater expression of avUCP mRNA (1.47-fold in PM, and 2.41-fold in TA) and avPGC-1α mRNA (1.42-fold in PM, and 2.08-fold in TA). In conclusion, the upregulation of avUCP and avPGC-1α reduced ROS accumulation and lipid oxidation at 45 min postmortem in the skeletal muscles of broilers stunned with hypercapnic moderate oxygenation GS. However, these changes were not sufficient to cause a difference in meat lipid oxidation at 24 h postmortem between broilers stunned with hypercapnic moderate oxygenation GS and those stunned with low-current, high-frequency ES.

Effects of dietary oxidized oil on laying performance, lipid metabolism, and apolipoprotein gene expression in laying hens.

We studied the effects of dietary oxidized oils on serum lipid metabolic indices, estradiol level, and the gene expression of apolipoprotein B-100 and apolipoprotein VLDL-II in laying hens. Hy-Line Grey hens (280 ± 10 d old; average egg production, 90.0 ± 2.5%) were allotted to 1 of 4 dietary treatments, which were supplemented with 0 (control group), 1% (low oxidized group), 2% (moderately oxidized group), or 4% (highly oxidized group) thermally oxidized soybean oil. Each treatment contained 6 replicates, with 12 birds each. The feeding trial lasted for 30 d. Laying performance data were recorded weekly. Other indices were measured on d 0, 2, 6, 14, and 30 of the feeding trial. Hens in the moderately and highly oxidized groups had significantly lowered feed conversion ratios (P < 0.05). Those in the highly oxidized group also had decreased concentrations of serum very low density lipoprotein cholesterol on d 30 (P < 0.05) compared with the very low density lipoprotein cholesterol of hens in the moderately oxidized group. Hens in the moderately oxidized group had significantly increased expression of apolipoprotein B-100 (P < 0.05) from d 6 to 30. Consequently, hepatic triglyceride increased in this group on d 30 (P < 0.05). Serum triglyceride decreased in the moderately oxidized group on d 30 (P < 0.05), which may have been caused by the activation of peroxisome proliferator-activating receptor α. Serum estradiol levels were not significantly affected by oxidized oils at any time of measurement, but were significantly different between d 0 and 30 within the moderately oxidized group. This fact indicated that the effect of oxidized oils on apolipoprotein B-100 might partially be a cumulative result of the increasing secretion of estradiol. The results suggested that oxidized oil may affect the performance of laying hens through the regulation of apolipoproteins and estradiol.

Comparison of blood variables, fiber intensity, and muscle metabolites in hot-boned muscles from electrical- and gas-stunned broilers.

The aim of this study was to compare the effects of gas stunning (GS) and electrical stunning (ES) on energy metabolism in Arbor Acres broilers. Thirty-six birds were slaughtered without stunning (control) or after stunning with the following treatments: 40% CO(2) + 21% O(2) + N(2) (G40%); 60% CO(2) + 21% O(2) + N(2) (G60%); 35 V, 47 mA, 400 Hz (E35V); 50 V, 67 mA, 160 Hz (E50V); and 65 V, 86 mA, 1,000 Hz (E65V). Muscle samples were obtained from the pectoralis major (breast) and tibialis anterior (leg) muscles in ambient temperature within 45 min postmortem and stored at -80°C. Blood pH decreased consistently with GS (G40% and G60%) compared with ES and the control (P < 0.01). No consistent differences were observed between GS and ES in the plasma variables, glycolytic potential, adenosine phosphates, or fiber intensities. Plasma lactate increased with G40% and E35V (P < 0.05), whereas plasma uric acid and urea nitrogen increased with E35V (P < 0.05) compared with the control. Compared with the control, the intensity of type IIB fibers decreased in broilers stunned with E35V and E50V (P < 0.05) and glycolytic potential increased (P < 0.01) with G60% in the breast muscle and decreased (P < 0.01) in the leg muscle with all the stunning treatments except for E50V. Energy decreased (lower adenosine triphosphate, higher adenosine monophosphate, and adenosine monophosphate:adenosine triphosphate ratio, P < 0.05) in breast muscle with G40% compared with ES at high currents (E50V and E65V). However, the adenosine phosphates with GS were not significantly different (P > 0.05) from ES at low current (E35V) in either breast or leg muscle. In conclusion, no essential difference in energy metabolism was found in broilers stunned with ES and GS when ES was based on low current and high frequency and GS was based on hypercapnic moderate oxygenation. This study indicated that G40% was potentially a superior stunning variable.

Transport stress in broilers. II. Superoxide production, adenosine phosphate concentrations, and mRNA levels of avian uncoupling protein, avian adenine nucleotide translocator, and avian peroxisome proliferator-activated receptor-gamma coactivator-1alpha in skeletal muscles.

The effect of transport stress on superoxide production and adenosine phosphate concentration in addition to avian uncoupling protein (avUCP), avian adenine nucleotide translocator, and avian peroxisome proliferator-activated receptor-gamma coactivator-1alpha mRNA levels of skeletal muscles in broilers was investigated. Arbor Acres chicks (n = 360, 46 d old, males) were randomly allotted to 1 of 5 treatments: unstressed control, 45-min (short-term) transport with 45-min (short-term) recovery, 45-min transport with 3-h (long-term) recovery, 3-h (long-term) transport with 45-min recovery, and 3-h transport with 3-h recovery. Each treatment consisted of 6 replicates with 12 birds each. All birds (except control group) were transported according to a designed protocol. Transport time affected reactive oxygen species production in the thigh muscle (P < 0.05), adenosine triphosphate (ATP) content and energy charge (EC) in both breast and thigh muscles (P < 0.05 for all 4 comparisons), ATP:adenosine diphosphate (ADP) ratio in the breast muscle (P < 0.05), and avUCP mRNA levels in the thigh muscle (P < 0.05). Long-term transport increased (P < 0.05) reactive oxygen species production, ATP content, ATP:ADP ratio, and EC in the thigh muscle, but it decreased ATP content, ATP:ADP ratio, and EC in the breast muscle. Long-term transport increased avUCP mRNA in the thigh muscle (P < 0.05). Long-term recovery increased the ATP (P < 0.05) and ADP (P < 0.05) concentrations, avian adenine nucleotide translocator mRNA (P < 0.05), and avian peroxisome proliferator-activated receptor-gamma coactivator-1alpha mRNA (P < 0.05) in the thigh muscle, whereas EC decreased (P < 0.05) in the breast muscle. There were interactions between transport and recovery time on ATP (P < 0.05), EC (P < 0.05), and avUCP mRNA level (P < 0.05) in the thigh muscle. This study suggests that long-term transport accelerates muscle energy metabolism and lipid peroxidation. A long-term recovery may help alleviate cellular damage and maintain meat quality by reducing the rate of energy metabolism and scavenging of free radicals formed.

Frequent epidermal growth factor receptor gene mutations in malignant pleural effusion of lung adenocarcinoma.

Malignant pleural effusions (MPEs) are often observed in lung cancer, especially adenocarcinoma. Epidermal growth factor receptor (EGFR) gene mutations are usually detected in lung adenocarcinoma. The purpose of the present study was to investigate the EGFR mutation rate in MPEs of lung adenocarcinoma. Between June 2005 and December 2006, 136 MPEs from lung adenocarcinoma were collected for EGFR mutation detection. In addition, between April 2001 and November 2004, 91 surgically resected specimens of lung adenocarcinoma from patients without MPEs were assessed for EGFR mutation. The EGFR mutation rate was significantly higher in the patients with MPEs than in the patients without (68.4% versus 50.5%). The EGFR mutation rate in patients with MPEs was not associated with sex, smoking history, age or cancer stage. By multivariate analysis, an age of <65 yrs, never smoking, Eastern Cooperative Oncology Group performance status 0-1, and EGFR mutation were significantly associated with a longer overall survival for lung adenocarcinoma patients with MPEs. The patients with malignant pleural effusions related to lung adenocarcinoma had a higher epidermal growth factor receptor gene mutation rate than the patients from whom surgically resected specimens were taken. Epidermal growth factor receptor tyrosine kinase inhibitors may be the treatment of choice for lung adenocarcinoma with malignant pleural effusions in east Asia.