A Phase II Study of Neoadjuvant Opnurasib KRAS G12C Inhibitor in Patients With Surgically Resectable Non-Small Cell Lung Cancer (CCTG IND.242A): A Substudy of the IND.242 Platform Master Protocol.

Molecularly targeted agents are increasingly being studied in the treatment of early-stage non-small cell lung cancer (NSCLC) to try and improve cure. However, phase 3 data on neoadjuvant therapy have largely been conducted in a biomarker agnostic manner with inconsistent exclusion of EGFR and ALK alterations. Our objective was to develop IND.242 as a large-scale neoadjuvant platform trial to introduce novel agents into the preoperative window for molecularly-defined NSCLC patient populations. Given that KRAS G12C mutations are common in the overall NSCLC patient population, ranging from 9.4% to 13% of cases across different cohorts, and may be associated to worse prognosis, the initial IND.242A Substudy was designed to test neoadjuvant JDQ443 (opnurasib), a selective KRAS G12C inhibitor. This current trial report describes the multicenter, Canadian Cancer Trials Group (CCTG)-led IND.242 neoadjuvant phase 2 platform master protocol and its first Substudy (IND.242A) of neoadjuvant opnurasib KRAS G12C inhibitor for patients with surgically resectable NSCLC (AJCC 8th edition stage IA2 to IIIA). In IND.242A, a maximum of 27 patients will be accrued in participating Canadian sites. The primary objective is the rate of major pathological response (MPR) following neoadjuvant opnurasib. Secondary objectives include safety and tolerability of the treatment regimen, objective response rate (ORR) by RECIST 1.1 for the neoadjuvant treatment period, pathological complete response (pCR) rate, event-free survival (EFS) at 2 years, and surgical outcomes. Exploratory objectives are to explore patient related outcomes (PROs) and identify potential predictive biomarkers of response and mechanisms of resistance on tissue and peripheral blood samples.

Dietary metabolizable energy and crude protein levels affect pectoral muscle composition and gut microbiota in native growing chickens.

The experiment aimed to study effects of dietary metabolizable energy (ME) and crude protein (CP) levels alone and in interaction on performance, pectoral muscle composition and gut microbiota in native growing chickens. A total of 648 10-wks-old Beijing-You Chicken (BYC) female chickens were randomly allocated to 9 groups with 6 replicates per group and 12 chickens per replicate, and the chickens were fed with a 3 × 3 factorial diets (3 levels of dietary ME: 11.31 MJ/kg, 11.51 MJ/kg, 11.71 MJ/kg; and 3 levels of dietary CP: 14%, 15%, 16%). The results showed that dietary ME and CP levels didn't affect average feed intake (AFI), body weight gain, feed gain ratio (P > 0.05), but ME level significantly affected the AFI (P < 0.05); mortality rate of 11.31 MJ/kg group was the highest (P < 0.05). Dietary ME, CP levels, and the interaction significantly affected pectoral CP and crude fat (CF) content of the growing chickens (P < 0.01). Dietary CP level had opposite effects on pectoral CP and CF content (P < 0.01). The 16% CP increased the pectoral CF content, which may have a negative impact on meat flavor. Dietary ME level affected 11 types of pectoral free amino acids (FAA) contents, including aspartic acid, L-threonine (P < 0.05), also amino acid classification, for example, total amino acid (TAA) and essential amino acid (EAA) content (P < 0.05). The 11.51 MJ/kg group had the highest TAA, EAA, delicious amino acid (DAA) content and EAA percentage (P < 0.05), while 11.31 MJ/kg group had the lowest bitter amino acid (BAA) content and BAA percentage and the highest fresh and sweet amino acid (FSAA) percentage (P < 0.05). Dietary CP level significantly affected glutamine and tyrosine content (P < 0.05). The interaction of dietary ME and CP level affected C20:3n6 content, saturated fatty acid (SFA), and unsaturated fatty acid (UFA) percentage (P < 0.05). The CP level significantly affected SFA percentage (P < 0.05). The 16% CP level increased the diversity of gut microbiota, but at the same time increased the relative abundance of Proteobacteria (P < 0.05), which is a sign of microbiota disorder. The increase of dietary ME level resulted in a gradual decrease in the diversity and relative abundance of gut microbiota. In conclusion, the present study suggested that the medium dietary ME (11.51 MJ/kg) and low CP (14-15%) levels can be helpful for enhancing pectoral muscle composition, increase meat quality such as flavor and nutritional value, and benefit for gut microbiota in native growing chickens.

Dietary calcium and non-phytate phosphorus levels affect the performance, serum biochemical indices, and lipid metabolism in growing pullets.

This experiment aimed to study the effects of dietary calcium (Ca) and non-phytate phosphorus (NPP) levels on performance, serum biochemical indices, and lipid metabolism in Beijing You Chicken (BYC), a local chicken. A 3 × 3 factorial design was adopted, dietary Ca levels were 0.66, 0.71, and 0.76%, NPP levels were 0.25, 0.30, and 0.35%. A total of 648 ten-wk-old BYC growing pullets were randomly divided into 9 groups with 6 replicates per group, and 12 birds per replicate. Growth performance, serum biochemical indices, and lipid metabolism indicators from 10 to 16 wk were measured. The results showed as follows: 1) Dietary Ca and NPP alone did not affect growth performance, but the interaction of dietary Ca and NPP affected average feed intake (AFI) of growing pullets (P < 0.05). The AFI was the lowest for the group with 0.71% Ca and 0.25% NPP (3,550.0 g, P = 0.036). 2) Dietary Ca level significantly affected serum P content (P < 0.05); dietary NPP had an influence trend on serum Ca content (P= 0.054). Dietary NPP levels and the interaction of Ca and NPP significantly affected alkaline phosphatase (AKP) activity. 3) Dietary Ca levels significantly affected TC content and HDL-C content (P < 0.05). Dietary NPP level significantly affected TG content (P < 0.05), the TG content in 0.25% and 0.30% NPP groups was significantly lower than that in 0.35% NPP group (P < 0.05). The interaction of dietary Ca and NPP significantly affected TG, TC and HDL-C contents (P < 0.05). TG, TC, and LDL-C levels were lower and HDL-C levels were the highest in the group with 0.66% Ca and 0.25% NPP. In summary, appropriate dietary Ca level can regulate serum TG, TC, and HDL-C content. Dietary Ca and NPP levels can be adjusted in pullet phase to avoid excessive obesity during the egg-laying period. This study recommended that dietary 0.66% Ca and 0.25% NPP benefit for the lipid metabolism of BYC growing pullets without affecting the performance.

Lighting pattern and photoperiod affect the range use and feather cover of native laying hens under free range condition.

The study aimed to investigate the effects of lighting pattern and photoperiod on range use, feather cover and gait score of native laying hens. Six hundred and thirty 19-wks-old Beijing You Chicken (BYC) pullets were randomly allocated to 6 groups with 105 birds each, 3 replicates per group, reared in individually lit indoor pens with separate covered shed and outdoor areas. A 2 × 3 factorial experiment (2 lighting patterns: continuous and intermittent lighting; 3 photoperiods: 16 h, 14 h, 12 h) was arranged indoors, including 16L:8D (6:00-22:00) for group 1; 12L:2D:4L:6D (6:00-18:00,20:00-24:00) for group 2; 14L:10D (6:00-20:00) for group 3; 10L:2D:4L:8D (6:00-16:00,18:00-22:00) for group 4; 12L:12D (6:00-18:00) for group 5, and 8L:4D:4L:8D (6:00-14:00,18:00-22:00) for group 6, respectively. The number of hens in indoor pen, covered shed and outdoor area were counted at 8:00, 9:00, 10:00, 11:00, 12:00, 13:00, 14:00 for consecutive 3 sunny days at 34 wks and 36 wks of age, feather cover and gait score of the laying hens were assessed at the end of 36 wks. The results showed that daily average hen percentage was the highest in continuous 12 h group (63.67%), and the lowest in intermittent 14 h group (58.36%) in indoor pen (P < 0.05); the daily average hen percentage was the lowest in continuous 12 h group (16.05%), and the highest in intermittent 14h group (21.22%) in outdoor area (P < 0.05). Lighting pattern significantly affected hen percentage in indoor pen and outdoor area, the hen percentage in indoor pen was higher in continuous lighting groups than in intermittent lighting groups (62.09% vs. 59.23%) (P < 0.05), the hen percentage in outdoor area was lower in continuous lighting groups than in intermittent lighting groups (16.60% vs. 19.95%) (P < 0.05). Photoperiod had no effect on the hen percentage (P > 0.05), but time of day affected the hen percentage in different areas (P < 0.05). The feather cover score was higher in intermittent lighting groups than in continuous lighting groups (17.43 vs. 15.04, P < 0.05). The average hen percentage in indoor pen is strongly negatively correlated with the feather cover score (r = -0.880, P = 0.050), and the hen percentage in outdoor area is strongly positively correlated with the feather cover score in intermittent lighting condition (r = 0.811, P < 0.05). The present study suggested that more range use is beneficial for the feather cover and physical health of laying hens under free range condition, and intermittent lighting is more conducive to range use and feather cover of native laying hens, which may be related to its affecting hens' rhythmic activities, increasing adaptation to outdoor environment, and reducing the incidence of feather pecking and parasites.

[Incidence and risk factors of thromboembolism in patients with lung cancer receiving immunotherapy].

Objective: To evaluate the incidence of thromboembolism in a cohort of patients with lung cancer who received immune checkpoint inhibitors (ICIs), and explore relevant clinical risk factors. Methods: We retrospectively collected and analyzed the clinical data of patients with confirmed primary lung cancer and treated with ICIs between March 2018 and June 2021 at three hospitals in China (Tongji Hospital of Tongji Medical College of Huazhong University of Science and Technology, Fudan University Shanghai Cancer Center and Zhongshan Hospital of Fudan University). The cumulative incidence and risk factors of thromboembolism in these patients were analyzed using a competitive risk model. Results: A total of 804 patients were enrolled, there were 623 males and 181 females, with a median age of 59 years (ranged 28-86 years). Of these, 62 patients encountered 65 thromboembolic events, including 51 venous thromboembolism events (VTE) and 14 arterial thromboembolism events. The cumulative incidence of thromboembolism events at 3, 6, 12 and 24 months were 4.3%, 6.1%, 10.1% and 16.8%, respectively. And the cumulative incidence of venous thromboembolism events at 3, 6, 12 and 24 months were 3.4%, 4.7%, 9.0% and 13.3%, respectively. Multivariate analysis showed that history of thromboembolism (HR=6.345, 95%CI: 2.917-13.802,P<0.001),liver metastasis (HR=2.249, 95%CI: 1.123-4.502,P=0.022) and peripherally inserted central venous catheter (HR=3.674, 95%CI: 1.751-7.712, P<0.001) were independent risk factors for venous thromboembolism during ICIs therapy in patients with lung cancer. Conclusions: Patients with lung cancer under ICIs therapy are at high risk of thromboembolism. And history of thromboembolism, liver metastasis and peripherally inserted central venous catheter are risk factors of venous thromboembolism.

Effects of light regime on circadian rhythmic behavior and reproductive parameters in native laying hens.

The paper aimed to study the effects of light regime on circadian rhythmic behavior and reproductive parameters in a native laying hen, Beijing You Chicken (BYC) during 22 to 30 wk. A total of 630 19-wk-old BYC female chicken were allocated to 6 light regime groups with 3 replicates per group and 35 birds per replicate, reared in individually lit floor pens with separate outdoor areas. A 2 × 3 factorial experiment (2 lighting patterns: continuous and intermittent lighting; 3 photoperiods: 16 h, 14 h, 12 h) was arranged, including 16L:8D for group 1; 12L:2D:4L:6D for group 2; 14L:10D for group 3; 10L:2D:4L:8D for group 4; 12L:12D for group 5, and 8L:4D:4L:8D for group 6, respectively. The circadian rhythmic behavior of the hens, including feeding, egg-laying and sleeping behaviors were observed by scan sampling and target sampling method for consecutive 3 d every other week during 22 to 30 wk. Infrared cameras were set outside each pen to record the rhythmic behaviors of the birds every other hour (6:00-7:00;8:00-9:00;10:00-11:00;12:00-13:00;14:00-15:00;16:00-17:00;18:00-19:00;20:00-21:00;22:00-23:00;0:00-1:00;2:00-3:00;4:00-5:00). Theovarian weight, oviduct weight, oviduct length, the number of large yellow follicles (LYF), and small yellow follicles (SYF) were measured at the end of 30 wk. The egg-laying rate during 22 to 30 wk was measured. The results showed that the highest feeding frequency was given by the 16L:8D, and the lowest given by the 12L:12D (P < 0.05). The average feeding duration was the longest in the 8L:4D:4L:8D at 6:00 to 8:00 (18.67 min/hen) (P < 0.05), and the shortest in the 8L:4D:4L:8D at 18:00 to 20:00 (3.75 min/hen) (P < 0.05). The highest egg-laying frequency was given by the 8L:4D:4L:8D (0.28), the lowest given by the 10L:2D:4L:8D (0.21) (P < 0.05). The shortest egg-laying duration occurred at 6:00 to 8:00, was given by the 16L:8D (8 min/hen), and the longest egg-laying duration occurred at 18:00 to 20:00, was given by 16L:8D and 8L:4D:4L:8D (>20.5 min/hen) (P < 0.05). The highest sleeping frequency was given by the 12L:2D:4L:6D, the lowest was given by the 12L:12D (P < 0.05). At 6:00 to 8:00, the shortest sleeping duration was given by 12L:2D:4L:6D (1.5 s/hen), at 18:00 to 20:00, the longest given by 8L:4D:4L:8D (14.3 s/hen) (P < 0.05). Lighting pattern and photoperiod alone or in interaction had no effect on egg-laying rate during 22 to 30 wk (P > 0.05), but had significant effects on ovarian weight, oviduct weight, oviduct length, the number of LYF and SYF (P < 0.05), and the continuous groups were all significantly higher than the intermittent groups (P < 0.05). The present study indicated that light regimes affected the frequency and duration of circadian behavior in BYC laying hens, and the continuous light was more beneficial to the reproductive development than the intermittent light in the early laying period.

Consensus for HER2 alterations testing in non-small-cell lung cancer.

Human epidermal growth factor receptor 2 (HER2) is a transmembrane glycoprotein receptor with intracellular tyrosine kinase activity. Its alterations, including mutation, amplification and overexpression, could result in oncogenic potential and have been detected in many cancers such as non-small-cell lung cancer (NSCLC). Such alterations are, in general, considered markers of poor prognosis. Anti-HER2 antibody-drug conjugates, e.g. trastuzumab deruxtecan (T-DXd, DS-8201) and disitamab vedotin (RC48), were recently approved for HER2-positive breast and gastric cancers. Meanwhile, several HER2-targeted drugs, such as T-DXd, neratinib, afatinib, poziotinib and pyrotinib, have been evaluated in patients with advanced NSCLC, with several of them demonstrating clinical benefit. Therefore, identifying HER2 alterations is pivotal for NSCLC patients to benefit from these targeted therapies. Recent guidelines on HER2 testing were developed for breast and gastric cancer, however, and have not been fully established for NSCLC. The expert group here reached a consensus on HER2 alteration testing in NSCLC with the focus on clinicopathologic characteristics, therapies, detection methods and diagnostic criteria for HER2-altered NSCLC patients. We hope this consensus could improve the clinical management of NSCLC patients with HER2 alterations.

Effects of lighting regimes on performance, pineal melanopsin expression and melatonin content in native laying hens aged from 19 to 34 weeks.

Melanopsin, a key light sensitive pigment, plays an important role in the regulation of bio-rhythm and photo-adaptation in poultry. This study aimed to investigate the effects of different lighting regimes on performance, pineal melanopsin expression and melatonin content in a native chicken, Beijing You Chicken (BYC) aged from 19 to 34 wk. A total of 900 nineteen-wk-old BYC female chicken having no significant body weight differences were randomly allocated to 3 groups with 3 replicates each, 100 birds each replicate, reared in individually lit floor pens with separate outdoor areas. Three different lighting regimes were used, including continuous 16 h (16L:8D, 6:00-22:00) for group 1, intermittent 16 h (12L:2D:4L:6D, 6:00-18:00, 20:00-24:00) for group 2, and continuous 12 h (12L:12D, 6:00-18:00) for group 3, respectively. The performance was measured for 19 to 34 wk. Serum melatonin (Mel), prolactin (Prl), luteinizing hormone (LH), and 17-beta estradiol (E2) contents were measured at 24 wk, 29 wk, and 34 wk of age, the relative expression of pineal melanopsin gene (Opn4 mRNA) was measured on 1 d at 9:00, 13:00, 17:00, 21:00, 1:00, and 5:00 at 29 wk of age, and at the end of 29 wk and 34 wk. The results showed that the egg mass, egg-laying rate, and feed egg ratio of BYC were not affected by lighting regimes for 19 to 34 wk (P > 0.05), except for the average feed intake (AFI) (P < 0.05). The AFI in the 12L:12D group was significantly higher than that in the 16L:8D group (P < 0.05), but had no difference with that in the 12L:2D:4L:6D group. The pineal Opn4 mRNA level was significantly upregulated in the 12L:2D:4L:6D group and downregulated in the 12L:12D group when compared with 16L:8D group at 29 and 34 wks of age (P < 0.05). The Mel content in the 16L:8D group was lower than that in the other 2 groups at 29 wk of age (P < 0.05), there was no difference in Mel content between 16L:8D group and 12L:2D:4L:6D group at 34 wk of age (P > 0.05). The present study suggested that the pineal melanopsin expression of the birds in the intermittent 16 h lighting group was higher than in the continuous 16 h and 12 h lighting group, and a significant negative correlation was found between melanopsin expression and Mel content at 34 wk of age, which may interact to promote the photo-adaptation of the native chicken and affect the future laying performance.

Transcriptome profiling reveals morphogenesis-related candidate genes and pathways in the chick embryonic small intestine.

1. A better understanding of intestinal development is essential for the intestinal health of poultry. Intestinal villification starts on embryo day E15 and is generally completed before hatching (E21). The development of lymphoid organs in the intestine starts during embryogenesis. However, transcriptional information on the processing of intestinal morphogenesis and immune development during chick embryogenesis is limited.2. In this work, RNA-sequencing was performed using 12 biological replicates to investigate Hy-Line brown chick embryonic small intestinal transcription at E15 and E21. Differentially expressed genes (DEGs) between E15 and E21 were identified. GO and KEGG enrichment analyses, based on the DEGs, were performed to identify key GO terms in the biological process category and key KEGG pathways. PPI networks were constructed based on the DEGs in the key pathways to screen hub genes. The embryonic small intestinal morphology and IgA distribution were observed by histological processing. The serum levels of IgA and lysozyme were measured by ELISA.3. A total of 76.38 Gb of high-quality RNA-sequencing data were generated and uploaded. A total of 2,676 DEGs, between E15 and E21, were identified. Structural development and villification of the small intestine at E15 tended to proceed via the expression of nervous system development-related genes. A combination of the histological and serological results with the transcriptome data indicated that the identified genes and pathways may be strong candidates for intestinal morphogenesis-regulation.4. The small intestine appears to have developed a relatively complete morphology and transport, metabolism, digestion and immunity functions by E21. This work provided a transcriptome profile of the chick embryonic small intestine and provided insights into the intestinal development and health of poultry.

[Effect of dissipating phlegm and blood stasis simultaneously on AGEs/RAGE axis and oxidative stress in rats with diabetic myocardial microangiopathy].

OBJECTIVE: To investigate the effect of dissipating phlegm and blood stasis simultaneously for protecting cardiac microvascular endothelial cells (CMECs) against high glucose-induced injury and the role of AGEs/RAGE axis in the underlying mechanism. METHODS: The primary CMECs were isolated from rat heart by enzymatic digestion and identified by immunofluorescence assay. The CMECs exposed to 33 mmol/L glucose for 48 h were divided into model group (MC), resolving phlegm (RP) group, dissipating blood stasis (DBS) group, dissipating phlegm and blood stasis (RPDBS) group and ALT-711 group. After treatment with 10% drug-containing serum and ALT-711 for 48 h, the content of AGEs in the cells were measured with ELISA. The expressions of RAGE mRNA and protein were measured with real-time quantitative PCR, immunofluorescence assay and Western blotting; The activity of NADPH oxidase and ROS level were measured by cytochrome c reduction and fluorescent probe DHE. RESULTS: High glucose exposure significantly increased the content of AGEs, RAGE expressions at the protein and mRNA levels, NADPH oxidase activity and ROS level in the CMECs (P < 0.01). These changes were significantly mitigated by treatments with RP, DBS, RPDBS and ALT-711 (P < 0.01), among which RPDBS caused the most significant decrements in AGEs content, RAGE expression and NADPH oxidase activity (P < 0.01, P < 0.05). The reduction of ROS level in the RPDBS group was significantly greater than that in RP group (P < 0.01), but similar to that in DBS group (P > 0.05). CONCLUSION: Dissipating phlegm and blood stasis simultaneously can be helpful for prevention and treatment of diabetic myocardial microangiopathy by suppressing the excessive activation of AGEs-RAGE signal axis and oxidative stress, thus protecting CMECs against high glucose-induced damage. Dissipating phlegm and blood stasis simultaneously is better than either of the therapy alone.

Effects of light regime on the hatching performance, body development and serum biochemical indexes in Beijing You Chicken.

The paper aimed to study the effects of light regime during the incubation on hatching performance, body development and serum biochemical indexes in Beijing You Chicken (BYC). A total of 1,408 BYC eggs were randomly allocated into 4 groups: 24 h dark as the control (Inc24D); 3 light regimes were 8 h light and 16 h dark group (Inc8L:16D); 12 h light and 12 h dark group (Inc12L:12D); 16 h light and 8 h dark group (Inc16L:8D), respectively. There were 352 eggs in each group, 4 replicates each group and 1 tray each replicate, 88 eggs each tray. Light-emitting diode (LED) strips, white light with temperature of 4,500 to 5,000 K, 150 to 200 lx were set up. The results showed that light regimes had no significant effects on hatching rate of eggs, hatching rate of fertile eggs and healthy rate of chicks (P > 0.05), but the hatching rate of eggs was the lowest in the Inc24D group (87.22%), and the highest in the Inc12L:12D group (93.64%); Lighted incubation significantly affected the incidence of leg problems of 1-day-old chicks (P < 0.05). The incidence rate of leg problems was the highest in the Inc24D group (4.21%), and was decreased in Inc12L:12D and Inc16L:8D groups (P < 0.05). Femur length in the Inc12L:12D group was greater than that in the Inc24D and Inc16L:8D groups (P = 0.011), but there were no differences between Inc8L:16D group and other three groups (P > 0.05). The relative brain weight of 1-day-old chicks was higher in Inc24D and Inc16L:8D groups than in Inc8L:16D group (P = 0.052), but had no difference with Inc12L:12D group. Light regimes during incubation had no effects on serum total protein, albumen, globulin, and urea nitrogen content of 1-day-old chicks (P > 0.05), while the globulin content in Inc12L:12D group was numerically greater than in other three groups (P = 0.063). Lysozyme content in Inc12L:12D group was higher than that in the Inc24D and Inc8L:16D groups (P < 0.05), but had no difference with that in Inc16L:8D group. Light regime had no effect on serum total antioxidant capability (T-AOC) (P > 0.05), but significantly affected the activities of glutathione peroxidase (GSH-Px) and total superoxide dismutase (T-SOD) (P < 0.05). The GSH-Px and T-SOD activities in the Inc12L:12D group were higher than those in the Inc24D and Inc8L:16D groups. The MDA content of Inc24D was the highest (9.67 nmol/mL) compared to the others, which was very close to the significant level (P = 0.056). In conclusion, 12 h light and 12 h dark incubation has the potential to improve the hatching performance of BYC eggs, benefit for the long bone development, improve some serum immune and antioxidant indexes, and reduce the leg problems in 1-day-old chicks.

Selection and implementation of single nucleotide polymorphism markers for parentage analysis in crossbred cattle population.

Crossbreeding is an essential way of improving herd performance. However, frequent parentage record errors appear, which results in the lower accuracy of genetic parameter estimation and genetic evaluation. This study aims to build a single nucleotide polymorphism (SNP) panel with sufficient power for parentage testing in the crossbred population of Simmental and Holstein cattle. The direct sequencing technique in PCR products of pooling DNA along with matrix-assisted laser desorption/ionization time-of-flight MS method for genotyping the individuals was applied. A panel comprising 50 highly informative SNPs for parentage analysis was developed in the crossbred population. The average minor allele frequency for SNPs was 0.43, and the cumulative probability of exclusion for single-parent and both-parent inference met 0.99797 and 0.999999, respectively. The maker-set for parentage verification was then used in a group of 81 trios with aid of the likelihood-based parentage-assignment program of Cervus software. Reconfirmation with on-farm records showed that this 50-SNP system could provide sufficient and reliable information for parentage testing with the parental errors for mother-offspring and sire-offspring being 8.6 and 18.5%, respectively. In conclusion, a set of low-cost and efficient SNPs for the paternity testing in the Simmental and Holstein crossbred population are provided.

Effects of laying breeder hens dietary ß-carotene, curcumin, allicin, and sodium butyrate supplementation on the jejunal microbiota and immune response of their offspring chicks.

Our long-term goal is to improve chick health and reduce the use of antibiotics in the poultry industry via maternal effects. To link jejunal microbes with chicks' different immune levels and growth performance in our previous research, this study investigated jejunal microbes, jejunal inflammation, and immune responses based on a comparison between different groups. Newly hatched Hy-Line chicks were allotted into 3 groups: a chick control group (cCON), a ciprofloxacin lactate treatment group (Cipro)-the chicks of the cCON and Cipro groups were hatched from laying breeder hens given a basal diet-and a 5-wk ß-carotene, curcumin, allicin, and sodium butyrate supplementation group (cCCAB), wherein chicks hatched from laying breeder hens. All groups were fed the same diet for 4 wk; the Cipro group was given ciprofloxacin lactate in drinking water continuously. At the end of the experiment, the results demonstrated that the jejunal microbes of the Cipro group showed significant changes in alpha and beta diversity, and in taxonomy at phylum and genus levels. Statistically, a total of 67 significantly enriched (P < 0.05) taxa were identified between groups by linear discriminant analysis effect size; Firmicutes was significantly enriched (P < 0.05) in the cCCAB group, 65 taxa were significantly enriched (P < 0.05) in the Cipro group, and 32 of the 65 enriched (P < 0.05) taxa were in the Proteobacteria phylum of the Cipro group. Levels of lipopolysaccharide in jejunal content, and nuclear factor kappa-B, and tumor necrosis factor-α in jejunums of the Cipro and cCCAB groups were increased (all P < 0.05) compared to those in the cCON group. There was obvious neutrophil infiltration and upregulated (all P < 0.05) IL-6 mRNA in the Cipro group jejunums compared to the cCON and cCCAB groups. The expression of PSME3 and PSME4 genes was upregulated (all P < 0.05) in the cCCAB group compared to the cCON and Cipro groups. In conclusion, ciprofloxacin lactate administration led to potential hazards in health and growth in chicks via microbial disturbances-induced jejunal inflammation, and laying breeder hens dietary supplementation with ß-carotene, curcumin, allicin, and sodium butyrate could enhance jejunal immunity of their offspring via the interaction between host innate immunity selected microbial colonization and microbiota educated adaptive immunity.

Canadian consensus: a new systemic treatment algorithm for advanced EGFR-mutated non-small-cell lung cancer.

Background: Multiple clinical trials for the treatment of advanced EGFR-mutated non-small-cell lung cancer (nsclc) have recently been reported. As a result, the treatment algorithm has changed, and many important clinical questions have been raised:■ What is the optimal first-line treatment for patients with EGFR-mutated nsclc?■ What is preferred first-line treatment for patients with brain metastasis?■ What is the preferred second-line treatment for patients who received first-line first- or second-generation tyrosine kinase inhibitors (tkis)?■ What is the preferred treatment after osimertinib?■ What evidence do we have for treating patients whose tumours harbour uncommon EGFR mutations? Methods: A Canadian expert panel was convened to define the key clinical questions, review recent evidence, and discuss and agree on practice recommendations for the treatment of advanced EGFR-mutated nsclc. Results: The published overall survival results for osimertinib, combined with its central nervous system activity, have led to osimertinib becoming the preferred first-line treatment for patients with common EGFR mutations, including those with brain metastasis. Other agents could still have a role, especially when osimertinib is not available or not tolerated. Treatment in subsequent lines of therapy depends on the first-line therapy or on T790M mutation status. Treatment recommendations for patients whose tumours harbour uncommon EGFR mutations are guided mainly by retrospective and limited prospective evidence. Finally, the evidence for sequencing and combining tkis with chemotherapy, angiogenesis inhibitors, checkpoint inhibitors, and other new therapeutics is reviewed. Conclusions: This Canadian expert consensus statement and algorithm were driven by significant advances in the treatment of EGFR-mutated nsclc.

Effects of indoor stocking density on performance, egg quality, and welfare status of a native chicken during 22 to 38 weeks.

This study investigated the effects of indoor stocking density on performance, egg quality, and welfare status of a native chicken, Beijing You Chicken (BYC), during 22 to 38 wk. A total of 1,040 19-wk-old BYC pullets were randomly allocated to 4 groups (2 replicates each) and reared in 8 individual floor pens with separate covered shed and uncovered outdoor areas. The indoor stocking densities were 5, 6, 7, and 8 hens/m2, and the birds were fed corn-soybean based diets. The performance was calculated for 22 to 30, 30 to 38, and 22 to 38 wk, and egg quality indices were measured at the end of weeks 26, 29, and 36. The feather cover and gait score of the birds were assessed at the end of weeks 29 and 36. The results showed that average feed intake (AFI) and mortality rate of BYC in the 8 hens/m2 group were higher than other groups during 22 to 30 wk (P = 0.001 and P = 0.005); the egg mass and egg-laying rate were higher in groups with lower stocking density, in contrast to the feed egg ratio during 30 to 38 wk. The AFI, mortality rate, and 38-wk body weight were affected by stocking density during 22 to 38 wk (P < 0.05). Most of egg quality indices were not changed by stocking density (P > 0.05), except for individual indicators, such as Haugh unit at week 26 (P = 0.012) and egg grade at week 29 (P = 0.026). The feather cover and gait scores of birds were affected by indoor stocking density at 36 wk of age, with the 8 hens/m2 group having lower scores than the 5, 6, and 7 hens/m2 groups (P = 0.042 and P = 0.039), whereas the 7 hens/m2 groups having no difference with the 5 and 6 hens/m2 groups (P > 0.05). Overall, the results suggested that the performance and egg quality of BYC were not significantly affected by equal to or less than 7 hens/m2 under this system (P > 0.05), but 8 hens/m2 had an adverse effect on the performance and welfare status of the native chicken during 22 to 38 wk.

Clinical characteristics of COVID-19-infected cancer patients: a retrospective case study in three hospitals within Wuhan, China.

BACKGROUND: Cancer patients are regarded as a highly vulnerable group in the current Coronavirus Disease 2019 (COVID-19) pandemic. To date, the clinical characteristics of COVID-19-infected cancer patients remain largely unknown. PATIENTS AND METHODS: In this retrospective cohort study, we included cancer patients with laboratory-confirmed COVID-19 from three designated hospitals in Wuhan, China. Clinical data were collected from medical records from 13 January 2020 to 26 February 2020. Univariate and multivariate analyses were carried out to assess the risk factors associated with severe events defined as a condition requiring admission to an intensive care unit, the use of mechanical ventilation, or death. RESULTS: A total of 28 COVID-19-infected cancer patients were included; 17 (60.7%) patients were male. Median (interquartile range) age was 65.0 (56.0-70.0) years. Lung cancer was the most frequent cancer type (n = 7; 25.0%). Eight (28.6%) patients were suspected to have hospital-associated transmission. The following clinical features were shown in our cohort: fever (n = 23, 82.1%), dry cough (n = 22, 81%), and dyspnoea (n = 14, 50.0%), along with lymphopaenia (n = 23, 82.1%), high level of high-sensitivity C-reactive protein (n = 23, 82.1%), anaemia (n = 21, 75.0%), and hypoproteinaemia (n = 25, 89.3%). The common chest computed tomography (CT) findings were ground-glass opacity (n = 21, 75.0%) and patchy consolidation (n = 13, 46.3%). A total of 15 (53.6%) patients had severe events and the mortality rate was 28.6%. If the last antitumour treatment was within 14 days, it significantly increased the risk of developing severe events [hazard ratio (HR) = 4.079, 95% confidence interval (CI) 1.086-15.322, P = 0.037]. Furthermore, patchy consolidation on CT on admission was associated with a higher risk of developing severe events (HR = 5.438, 95% CI 1.498-19.748, P = 0.010). CONCLUSIONS: Cancer patients show deteriorating conditions and poor outcomes from the COVID-19 infection. It is recommended that cancer patients receiving antitumour treatments should have vigorous screening for COVID-19 infection and should avoid treatments causing immunosuppression or have their dosages decreased in case of COVID-19 coinfection.

Biopsy on progression in patients with EGFR mutation-positive advanced non-small-cell lung cancer-a Canadian experience.

Background: Epidermal growth factor receptor (egfr) tyrosine kinase inhibitors (tkis) are standard therapy for patients with advanced or metastatic non-small-cell lung cancer harbouring an EGFR mutation. Upon progression, 50%-60% develop a secondary T790M mutation. Recent trials demonstrated outcome improvement with osimertinib compared with standard platinum-based chemotherapy as second-line therapy for patients with secondary T790M mutation. To identify T790M, a biopsy of the tumour or, more recently, plasma is necessary. This retrospective study aimed to evaluate biopsy procedures and mutational analysis at 2 Canadian cancer centres. Methods: In a retrospective review of patients who were approached to enrol in the aura2, aura3, or astris studies, demographics, eligibility for rebiopsy upon progression after an egfr tki, rebiopsy methods and complications, number of rebiopsies, and incidence of the T790M mutation were collected. Results: Of 84 patients considered for trial enrolment, 80 signed a consent. In 78 patients who underwent rebiopsy, computed tomography or ultrasonography guidance were the most common methods used. The most common biopsy sites were lung and lymph nodes. The median number of rebiopsies performed to find a T790M mutation was 2. Only 9% of patients experienced complications. Of samples obtained, 74% were adequate for testing after initial rebiopsy. A T790M mutation was found in 47 patients, of whom 44 were enrolled on a trial. After multiple rebiopsies, only 5% of samples were inadequate for molecular analysis. Conclusions: In the Canadian setting, the acceptance of rebiopsy on progression was high. Multiple rebiopsies were clinically feasible and could increase the yield for T790M mutation. The incidence of complications was low despite the most common site for rebiopsy being lung.

Genomic origin and EGFR-TKI treatments of pulmonary adenosquamous carcinoma.

BACKGROUND: Adenosquamous carcinoma (ASC) of the lung is a heterogeneous disease that is composed of both adenocarcinoma components (ACC) and squamous cell carcinoma components (SCCC). Their genomic profile, genetic origin, and clinical management remain controversial. PATIENTS AND METHODS: Resected ASC and metastatic tumor in regional lymph nodes (LNs) were collected. The ACC and SCCC were separated by microdissection of primary tumor. The 1021 cancer-related genes were evaluated by next-generation sequencing independently in ACC and SCCC and LNs. Shared and private alterations in the two components were investigated. In addition, genomic profiles of independent cohorts of adenocarcinomas and squamous cell carcinomas were examined for comparison. We have also carried out a retrospective study of ASCs with known EGFR mutation status from 11 hospitals in China for their clinical outcomes. RESULTS: The most frequent alterations in 28 surgically resected ASCs include EGFR (79%), TP53 (68%), MAP3K1 (14%) mutations, EGFR amplifications (32%), and MDM2 amplifications (18%). Twenty-seven patients (96%) had shared variations between ACC and SCCC, and pure SCCC metastases were not found in metastatic LNs among these patients. Only one patient with geographically separated ACC and SCCC had no shared mutations. Inter-component heterogeneity was a common genetic event of ACC and SCCC. The genomic profile of ASC was similar to that of 170 adenocarcinomas, but different from that of 62 squamous cell carcinomas. The incidence of EGFR mutations in the retrospective analysis of 517 ASCs was 51.8%. Among the 129 EGFR-positive patients who received EGFR-TKIs, the objective response rate was 56.6% and the median progression-free survival was 10.1 months (95% confidence interval: 9.0-11.2). CONCLUSIONS: The ACC and SCCC share a monoclonal origin, a majority with genetically inter-component heterogeneity. ASC may represent a subtype of adenocarcinoma with EGFR mutation being the most common genomic anomaly and sharing similar efficacy to EGFR TKI.