Deep learning-based facial analysis for predicting difficult videolaryngoscopy: a feasibility study.

While videolaryngoscopy has resulted in better overall success rates of tracheal intubation, airway assessment is still an important prerequisite for safe airway management. This study aimed to create an artificial intelligence model to identify difficult videolaryngoscopy using a neural network. Baseline characteristics, medical history, bedside examination and seven facial images were included as predictor variables. ResNet-18 was introduced to recognise images and extract features. Different machine learning algorithms were utilised to develop predictive models. A videolaryngoscopy view of Cormack-Lehane grade of 1 or 2 was classified as 'non-difficult', while grade 3 or 4 was classified as 'difficult'. A total of 5849 patients were included, of whom 5335 had non-difficult and 514 had difficult videolaryngoscopy. The facial model (only including facial images) using the Light Gradient Boosting Machine algorithm showed the highest area under the curve (95%CI) of 0.779 (0.733-0.825) with a sensitivity (95%CI) of 0.757 (0.650-0.845) and specificity (95%CI) of 0.721 (0.626-0.794) in the test set. Compared with bedside examination and multivariate scores (El-Ganzouri and Wilson), the facial model had significantly higher predictive performance (p < 0.001). Artificial intelligence-based facial analysis is a feasible technique for predicting difficulty during videolaryngoscopy, and the model developed using neural networks has higher predictive performance than traditional methods.

[Effect of dyslipidemia on clinical outcome of infertility patients receiving donor eggs].

Objective: To explore the effect of dyslipidemia on the clinical outcome of intracytoplasmic sperm injection-embryo transfer (ICSI-ET) in infertility patients receiving donor eggs. Methods: A total of 118 patients were selected to receive egg donors and ICSI-ET at the First Affiliated Hospital of Nanjing Medical University between April 2007 and December 2020. According to the levels of triacylglycerol, serum cholesterol, high density lipoprotein (HDL), and low density lipoprotein, they were divided into dyslipidemia group (35 cases) and normal blood lipids group (83 cases). The influence of body mass index (BMI) and age was adjusted by 1∶1 propensity score matching, and the general condition and clinical outcome of the two groups were analyzed retrospectively. Finally, the relationship between lipid composition and clinical outcome was analyzed according to patients' age and BMI. Results: (1) Comparing the pre-matching dyslipidemia group with the normal blood lipids group, the BMI of the dyslipidemia group was significantly higher than that of the normal blood lipids group [(23.5±2.4) vs (22.4±2.7) kg/m2], and the embryo implantation rate was significantly lower than that of the normal blood lipids group [13.6% (8/59) vs 27.3% (36/132)], the differences were statistically significant (both P<0.05). (2) There were no significant differences in years of infertility, number of pregnancies, number of abortions, number of transplanted embryos, protocol of endometrial preparation, endometrial thickness on transplantation day and high quality embryo rate between the two groups, through propensity score matching (all P>0.05). The biochemical pregnancy rate [28.6% (10/35)], embryo implantation rate [13.6% (8/59)] and live birth rate [20.0% (7/35)] in dyslipidemia group were significantly lower than those in the normal blood lipids group (P<0.05). The clinical pregnancy rate was lower than that of the normal blood lipids group (P>0.05). (3) The results of stratified analysis showed that the level of HDL in the clinically non-pregnant group was significantly lower than that in the pregnant group in patients ≤ 35 years old [(1.5±0.3) vs (1.8±0.5) mmol/L; P<0.05]. In the overweight recipient patients, the level of HDL of the clinically non-pregnant group was lower than that of the pregnant group (P>0.05). Conclusions: Dyslipidemia significantly reduces the biochemical pregnancy rate, embryo implantation rate and live birth rate in patients with receiving donor eggs. Especially in patients aged ≤35 years old, the reduction of HDL is closely related to adverse pregnancy outcomes.

[Association between serum uric acid to creatinine ratio and metabolic syndrome based on community residents in Chashan town, Dongguan city].

Objective: To analyze the association between serum uric acid to creatinine ratio (SUA/Cr) and metabolic syndrome among community residents in Chashan town, Dongguan city. Methods: Participants were from the prospective cohort study of chronic diseases in natural populations in South China conducted in Chashan town, Dongguan city from 2018 to 2019. A total of 11 334 participants with complete data were included by using convenient sampling method. Demographic characteristics, lifestyle and health status were collected through questionnaire and physical examination. The venous blood of the subjects was collected to detect the levels of serum uric acid, creatinine and blood lipid. All participants were divided into four groups (Q1-Q4) according to the quartile of SUA/Cr level. The relationship between SUA/Cr and metabolic syndrome and its components (abdominal obesity, high triglyceride, low level of high density lipoprotein cholesterol, hypertension and abnormal glucose metabolism) were analyzed by using logistic regression model. Results: The mean age of 11 334 participants was (49.52±10.02) years. Male participants accounted for 44.2% (5 015/11 334). The prevalence of metabolic syndrome was 31.2% (3 532/11 334), and the level of SUA/Cr was 5.17±1.53. The prevalence of metabolic syndrome in group Q1-Q4 was 22.3% (631/2 834), 26.5% (752/2 833), 34.9% (988/2 833) and 41.0% (1 161/2 834), respectively. After adjusting for relevant confounding factors, the result of logistic regression model showed that compared with group Q1, the risk of metabolic syndrome in group Q2-Q4 was significantly higher, with OR (95%CI) values about 1.41 (1.23-1.60), 2.19 (1.93-2.49) and 3.01 (2.65-3.42) respectively. The risk of each component of metabolic syndrome in group Q2-Q4 was higher (Ptrend<0.001). The SUA/Cr level of participants with normal uric acid level was significantly positively correlated with metabolic syndrome. The risk of metabolic syndrome increased with the increase of SUA/Cr level, but there was the same trend without significant differences in patients with hyperuricemia (Pinteraction=0.008). Conclusion: There is a positive correlation between SUA/Cr level and the risk of metabolic syndrome among community residents in Chashan town, Dongguan city.

[Relationship of quality and duration of sleep with hypertension among adults in Guangzhou].

Objective: To explore the relationship of sleep quality and sleep duration with hypertension among adults aged 30-79 years old in Guangzhou. Methods: According to multi-stage stratified cluster sampling, 12 747 residents aged 30-79 years old were sampled and surveyed in Guangzhou from January 2018 to March 2019. Data on general demographic characteristics, sleep quality, sleep duration and hypertension were collected through questionnaire survey, the Pittsburgh sleep quality index (PSQI) and physical examination. Multivariate logistic regression models were used to analyze the putative association between sleep quality, sleep duration and hypertension. Restrictive cubic spline curve was used to draw the dose-response relationship curve between sleep quality, sleep time and hypertension. Results: The mean age of the subjects was (52.68±12.17) years, the prevalence of hypertension was 36.6% (4 664/12 747), the average score of PSQI was (4.70±2.88), and the average sleep time was (7.00±1.32) hours. The prevalence of hypertension was positively associated with the PSQI score. Compared to the subjects with a score less than 3, OR (95%CI) of hypertension with a PSQI score of 3-5, 5-8, ≥9 were 1.14 (1.02-1.27), 1.17 (1.03-1.34), 1.41 (1.21-1.64), respectively. The relationship between sleep duration and hypertension appeared U-shaped. Compared with 6 to 8 hours sleep duration, both sleep duration<6 hours with OR(95%CI) of 1.27(1.12-1.43) or >8 hours with OR(95%CI) of 1.20(1.05-1.38) was associated with hypertension. Conclusion: Both poor sleep quality, longer or shorter sleep duration were responsible for increased risk of cognitive impairment in older Chinese.

The synergy of resveratrol and alcohol against Helicobacter pylori and underlying anti-Helicobacter pylori mechanism of resveratrol.

AIMS: To determine individual antibacterial and synergistic antibacterial effects of resveratrol and alcohol against Helicobacter pylori 26695 in vitro, and to elucidate the underlying mechanism of action of resveratrol against H. pylori. METHODS AND RESULTS: The minimum inhibitory concentrations (MICs) and time-killing curve of resveratrol and alcohol were determined. Transcriptome analysis by RNA sequencing was used to elucidate the underlying mechanism of action of resveratrol against H. pylori. Our results showed that the MICs of resveratrol and alcohol against H. pylori 26695 are about 64 µg ml-1 and 4% (v/v) respectively. The synergy was found: resveratrol at concentration of 64 µg ml-1 in combination with alcohol at concentration of 4% (v/v) showed >10 000-fold decrease in the mount of viable bacteria compared with resveratrol and alcohol used alone. Transcriptome analysis showed 152 genes were downregulated and 111 genes were upregulated in the presence of resveratrol. Genes involved in protein translation (17·1%), outer membrane proteins (OMPs) (9·9%) and transports (11·2%) comprise 38·2% of the downregulated genes. In comparison, genes involved in redox (13·5%), pathogenesis and motility (9·9%) and iron homeostasis (4·5%) comprise 27·9% of the upregulated genes. CONCLUSIONS: The synergy of resveratrol and alcohol against H. pylori was found in this study. The underlying mechanism of action of resveratrol against H. pylori may be mainly attributed to its inhibitory effect on translation, OMPs, transports, ATP synthase and possible oxidative damage. SIGNIFICANCE AND IMPACT OF THIS STUDY: Our study provides a global insight into the anti-H. pylori mechanism of resveratrol. Both resveratrol and alcohol can contribute to inhibition of ribosomes, changes in OMPs and oxidative damage, which may be the explanations of synergistic effect against H. pylori elicited by resveratrol and alcohol.

[Analysis of prognosis and pregnancy outcomes of fertility-preserving treatment for patients with stage â a, grade 2 endometrial cancer].

Objective: To investigate the efficacy and pregnancy outcome of fertility-preserving treatment for patients with stage Ⅰa, grade 2 endometrial cancer (EC). Methods: Clinical data was retrospectively collected for EC or atypical endometrial hyperplasia (AEH) patients treated in Peking University People's Hospital, Foshan First People's Hospital of Guangdong Province and First Affiliated Hospital of Sun Yat-sen University, from 2010 to 2019. Inclusion criteria for fertility-preserving treatment included: (1) Age ≤45 years. (2) EC with histological differentiation of G(1), G(2) or endometrial AEH. (3) EC disease should be stage Ⅰa, confined to the endometrium without myometrial invasion, lymph node or extrauterine metastasis. Treatment regimen: patients were given oral progestin therapy and endometrial pathology was evaluated every three months. Patients were divided into three groups as G(2) EC group, G(1) EC group and AEH group based on the histological differentiation. Oncological and pregnancy outcomes were compared among them. Results: (1) Totally 57 eligible patients were included in this study, including 11 cases with G(2) EC, 22 cases with G(1) EC, and 24 cases with AEH. (2) Oncological outcome: among the three groups of G(2) EC, G(1) EC and AH, the complete remission rates (9/11, 91% and 96%, respectively) and recurrence rates (3/9, 30% and 22%, respectively) were not significantly different (all P>0.05). Median remission time was significantly longer in the G(2) EC group than those in the other two groups (8, 6 and 4 months; P=0.046). Among 9 G(2) EC patients who recurred after complete remission, three patients relapsed at 7, 18 and 53 months, respectively. All 3 patients chose fertility-sparing treatment again, and all achieved complete remission after retreatment. (3) Pregnancy outcome: among the three groups, the assisted reproduction technology rates (4/8, 5/18 and 36%, respectively) and pregnancy rates (6/8, 5/18 and 36%, respectively) had no significant difference (P>0.05). However, time interval to pregnancy was shorter in G(2) EC patientsthan the other two groups (4, 9 and 22 months, respectively; P=0.006). Conclusions: Fertility-preserving treatment for patients with stageⅠa, G(2) endometrial cancer, may obtain a relatively high remission rate and an acceptable pregnancy rate. However, further exploration is needed due to the limited number of cases.

[Gait and neuromuscular activity changes in female older adults with knee osteoarthritis].

From November to December of 2018, twenty 65-year-old or older women patients with knee osteoarthritis were recruited from the Department of Physical Therapy, the First Affiliated Hospital of Anhui Medical University. Meanwhile, twenty healthy 65-year-old or older women were recruited from the local community. The results showed that the knee contact angle of the patient group was more flexed (P=0.040), and the minimum angle of the knee joint increased (P=0.008) during the stance period compared to the healthy group. However, there was no significant difference in the maximum contact angle between the angle of hip and ankle joints. In addition, the tibialis anterior muscle of the patients was significantly smaller than the healthy group (P=0.023). Therefore, knee osteoarthritis could change the gait and muscle activity of older women, especially the knee joint.

[Clinical analysis of 5 cases of acute poisoning by inhalation of hydrochlogen chloride].

Objective: To analyze the clinical characteristics caused by acute poisoning by inhalation of hydrogen chloride (HCl) and to raise awareness and treatment level of the disease. Methods: The clinical manifestations, imaging features, diagnosis, treatment and prognosis of 5 patients with acute HCl poisoning were analyzed retrospectively. Results: Among the 5 cases of HCl poisoning, 2 cases were severe poisoning, 3 cases were moderate poisoning. All patients were treated with corticosteroids and symptomatic treatment, one of them was treated with venovenous extracorporeal membrane oxygenation (VV-ECMO) . All patients were recovered and discharged from hospital. Conclusion: The lung damage of acute poisoning by inhalation of HCl is rapidly progressing, early detection and timely medical treatment can obtain a better prognosis.

[Application of semi-quantitative risk assessment method in occupational health risk assessment of wooden furniture manufacturing enterprises].

Objective: To explore the applicability of semi-quantitative risk assessment methods in wood furniture manufacturing companies. Methods: Two medium-sized wooden furniture manufactures were randomly selected as research objects, namely A company and B company. Used the Semi-Quantitative risk assessment method (the ratio method, the index method and the comprehensive method) in the "Guidelines for occupational health risk assessment of chemicals in the workplace" (GBZ/T 298-2017) to conduct occupational hygiene survey, occupational hazard factor testing and occupational health risk assessments for two wood furniture manufacturers from January to October 2018, and compared and analyzed the applicability of these three semi-quantitative risk assessment methods in wooden furniture manufacturing enterprises. Results: The occupational health status of A enterprises was worse than that of B enterprises, and the occupational health risk level was higher than that of B enterprises, and the risk level of A enterprise is 3~4, and the risk level of B enterprise is 2~3. The occupational health risk level obtained by the index method was consistent with the comprehensive method, while the risk level of some occupational disease hazards used the ratio method was inconsistent with the results of the index method and the comprehensive method. Compared with the index method and the comprehensive method, when E/OEL<0.5 or E/OEL≥2, there might be a certain fluctuation in the occupational health risk level obtained by the ratio method. Conclusion: The semi-quantitative risk assessment is more objective, comprehensive and flexible in the application of occupational health risk assessment, and can assess the occupational health risk level of chemical poisons in wood furniture manufacturing enterprises.

Micro124-mediated AHR expression regulates the inflammatory response of chronic rhinosinusitis (CRS) with nasal polyps.

MicroRNAs represent a component of the innate immune responses that can restrain inflammatory signaling, miR124 is an important member of inflammation-associated miRNAs, and abnormal miR124 expression is observed in many inflammatory diseases and immune disorders. However, the role and signaling pathways of miR124 in chronic rhinosinusitis with nasal polyps (CRSwNPs) have not been studied in detail. The aryl hydrocarbon receptor (AHR) is a ligand-inducible transcription factor that is highly conserved in evolution and plays important roles in the inflammatory response process. In our study, we describe the role of miR124 in the inflammatory response of CRS with nasal polyps. We found that the expression of miR124 was decreased in nasal polyps, and negatively correlated with the expression of AHR. MiR124 can inhibit AHR expression by directly target 3' untranslated region (3'-UTR) of AHR. To further investigate the relationship between miR124, AHR and CRS inflammatory response, we transfect HNEpC cells with miR124 mimic, miR124 inhibitors or siRNA of AHR, then all the results showed that miR124 could regulates cellular inflammatory response through negatively regulating AHR expression. This study demonstrated that the regulation of AHR expression by miR124 is critical to the development of inflammatory response in CRSwNPs.

Genetic variants in IL15 promoter affect transcription activity and intramuscular fat deposition in longissimus dorsi muscle of pigs.

Intramuscular fat (IMF) content is a key aspect of pork quality. Elucidation of intramuscular adipocyte regulation mechanisms is important for improving IMF content. Intramuscular adipocytes are dispersed among muscle fibers, so they are inclined to be affected by muscle-derived factors. Interleukin-15 is a major muscle-secreted factor. In this study, the genetic and physiological impacts of IL15 on adipogenesis is investigated. The promoter region of IL15 was scanned by comparative sequencing using two DNA pools of high- and low-IMF individuals. Two SNPs, c.-342C>T (ss2137497757) and c.-334G>A (ss2137497756) (the translation start site is designated as +1), were identified with reverse allele distribution in these two groups. Genotyping by allele-specific PCR revealed that the two SNPs were completely linked. The IMF content of TA/TA individuals was lower than that for CG/CG ones, whereas the IL15 expression level was higher in T-A/T-A individuals. Luciferase assaying also revealed that the T-A haplotype promoter had higher transcription activity. Meanwhile, the effect of interleukin-15 on adipocyte differentiation was further assessed in vitro. Results showed that interleukin-15 suppressed preadipocyte proliferation in a dose-dependent manner. The cell cycle of preadipocytes was arrested, and apoptosis was induced. Oil Red O staining and triglyceride quantification indicated that adipocyte differentiation was also inhibited by interleukin-15. The mRNA levels of PPARG and FABP4 decreased markably upon interleukin-15 treatment. Taken together, we identified two completely linked SNPs in the porcine IL15 promoter region that could alter IL15 transcription activity. As interleukin-15 can inhibit porcine adipocyte differentiation, these promoter mutations could affect IMF deposition by producing differential levels of muscle-derived interleukin-15.

Molecular cloning, alternative splicing and mRNA expression analysis of MAGI1 and its correlation with laying performance in geese.

1. In a previous study, a laying-related single nucleotide polymorphism (SNP) Record-106975 (A-G) was found in the intron of Membrane-associated Guanylate Kinase with Inverted Domain Structure-1 (MAGI1) gene by restriction site-associated DNA sequencing. This study was performed to further identify the function of MAGI1 on laying performance in Yangzhou geese. 2. The complete MAGI1 cDNA sequence was cloned from the ovary of the Yangzhou goose. A total of 4 alternative splice variants were identified, and the characterisation of their nucleic acid and amino acid (AA) sequences were analysed. The AA sequence of goose MAGI1 was highly conserved with other avian species. 3. MAGI1 was ubiquitously expressed in 11 tested tissues, with greater amounts of mRNA in the granulosa cell layer and ovary. GG (n = 14) and GA (n = 6) genotypes of SNP Record-106975 showed significantly lower MAGI1 mRNA levels than those of the AA genotype. 4. Over-expression and knockdown experiments revealed that MAGI1 mRNA expression was associated with caspase-3 mRNA levels in granulosa cells, and may play a role in regulating apoptosis of granulosa cells.

[Study on the choice of medication time and with drawal time of intermittent endocrine therapy for prostate cancer].

Objective: To determine the best timing of discontinuance of intermittent endocrine therapy for prostate cancer (PCa). Methods: From February 2012 to September 2014, 120 PCa patients confirmed by biopsy at the Department of Urology Surgery, Beijing Shijitan Hospital Affiliated to Capital Medical University, were included. They were divided into high-, moderate- and low-risk groups according to EAU Guidelines in 2015. All patients received intermittent endocrine therapy. When the serum level of prostate-specific antigen (PSA) dropped to 0.2 µg/L, the medication was not discontinued until 3 months later for the observation group; however, the medication was immediately discontinued after reaching the above-mentioned threshold for the control group. When the PSA level during the period of medication discontinuance increased to 4 µg/L, another course of intermittent endocrine therapy was administered until the occurrence of the failure event. The Cox proportional regression model was used to compare the difference between the two groups in the androgen-independent signs, progression of PCa and severe adverse events. Result: After correction for multiple confounding factors, including severity grading before treatment, average duration of medication, average duration of medication discontinuance and the number of courses of therapy, it was found that the medication scheme adopted in the observation group was a protective factor against androgen-independent signs in the high-risk PCa patients (HR: 0.535, 95% CI: 0.458-0.612; P<0.001); it was also a protective factor against disease progression for the high- and moderate-risk patients (high-risk group, HR: 0.387, 95% CI: 0.297-0.477; P<0.001; moderate-risk group, HR: 0.697, 95% CI: 0.581-0.813; P<0.001). However, there was no significant difference in the incidence of severe adverse events among PCa patients treated by different medication schemes (HR: 1.003, 95% CI: 0.906-1.100; P=0.798). The medication scheme adopted in the observation group was an independent risk factor for severe adverse events in the low-risk PCa patients (HR: 1.489, 95% CI: 1.305-1.673; P<0.001). Conclusion: As compared with the scheme of immediately discontinuing the medication after reaching the indicator threshold, maintenance medication for another 3 months was beneficial for the high- and moderate-risk PCa cases. The maintenance medication could reduce the androgen-independent signs and disease progression, without increasing the risk of severe adverse events. However, for the low-risk patients, there was no major difference between the two medication schemes, immediate discontinuance and maintenance medication for another 3 months. Instead, further medication may bring about a higher risk of severe adverse events.

[Distribution of electroencephalograph power density in patients with severe obstructive sleep apnea during different sleep stages].

Objective: To investigate the variation of electroencephalograph(EEG) power density during different sleep stages in OSA for understanding of the mechanisms underlying the brain dysfunction in OSA as well as its earlier diagnosis and treatment. Methods: Sixteen-channel EEGs from OSA patients and normal controls in stage wake, sleep stage 1, sleep stage 2, sleep stage 3 and rapid eye movement stage were analyzed by time-frequency analysis method. The EEG power density in different frequency bands (including δ, θ, α, σ, ß and γ) was respectively compared between the 2 groups. The correlation between the variation in the EEG power and primary indices of polysomnography was further analyzed. Results: The EEG power density in δ band in stage wake [OSA: (0.82±0.13) µV(2)/Hz, Control: (0.66±0.02) µV(2)/Hz, t=4.309, P<0.05], stage 1 [OSA: (1.28±0.07) µV(2)/Hz, Control: (0.92±0.04) µV(2)/Hz, t=-3.369, P<0.05] and stage 3 [OSA: (2.74±0.22) µV(2)/Hz, Control: (2.04±0.07) µV(2)/Hz, t=-2.669, P<0.05] was significantly higher in OSA, compared with that in the control. Statistical analysis showed that the EEG power density was significantly higher in frontal and central regions in stage wake [frontal: OSA: (0.90±0.02) µV(2)/Hz, Control: (0.66±0.02) µV(2)/Hz, t=8.539, P<0.01; central: OSA: (1.15±0.06) µV(2)/Hz, Control: (0.72±0.02) µV(2)/Hz, t=6.669, P<0.01] and stage 1 [frontal: OSA: (1.23±0.03) µV(2)/Hz, Control: (0.99±0.03) µV(2)/Hz, t=5.983, P<0.01; central: OSA: (1.52±0.05) µV(2)/Hz, Control: (1.14±0.04) µV(2)/Hz, t=5.714, P<0.01], as well as central region in stage 3 [OSA: (3.24±0.17) µV(2)/Hz, Control: (2.71±0.08) µV(2)/Hz, t=2.707, P<0.05]. The correlation analysis showed that the power density in central region in stage 1 and stage 3 was positively correlated with arousal index (r=0.877 in stage 1, 0.656 in stage 3), implying that sleep fragmentation was closely related to the variation of EEG power density during nocturnal sleep in OSA. Conclusions: The feature stages for OSA are stage wake, stage 1 and stage 3. The EEG power density in OSA (δ band) was significantly higher than that in the control. The EEG power density in OSA and the control shows differences in frontal and central regions in stage wake and stage 1, as well as in central region in stage 3. The results indicate that low-frequency EEG power density giving priority to frontal area and central area has improved in severe OSA, which may be related to the neurologic deficits in corresponding brain areas.

Gene Expression Characteristics and Regulation Mechanisms of Superoxide Dismutase and Its Physiological Roles in Plants under Stress.

Superoxide dismutases (SODs) are key enzymes functioning as the first line of antioxidant defense by virtue of the ability to convert highly reactive superoxide radicals to hydrogen peroxide and molecular oxygen. SOD plays a central role in protecting plants against the toxic effects of reactive oxygen species generated during normal cellular metabolic activity or as a result of various environmental stresses. Our review focuses on the characteristics of expression of SOD genes, the mechanisms regulating expression of SOD genes at transcriptional, posttranscriptional, and translation levels, and their functional role(s) during development and in response to biotic or abiotic stresses. We propose two important research directions: studying SOD at the genome-wide or proteome-wide level, and improving plant stress tolerances by selecting varieties using transgenic technology.

Molecular characterization, alternative splicing and expression analysis of ACSF2 and its correlation with egg-laying performance in geese.

ACSF2 (encoded by acyl-CoA synthetase family member 2) belongs to the acyl-CoA synthetase (ACS) family, activating fatty acids by forming a thioester bond with CoA. In our previous study, a SNP residing in the intron of ACSF2 was identified to be linked to goose egg-laying performance. But the structure of goose ACSF2 as well as its role in reproduction remains unknown. In this study, we cloned and characterized ACSF2 in Yangzhou geese. A total of four alternative splice variants, designated as ACSF2-1, ACSF2-2, ACSF2-3 and ACSF2-4 respectively, were identified in the ovary. The coding regions of the four variants are 1770, 1692, 1599 and 1917 bp in length, respectively encoding 589, 563, 532 and 638 amino acids with conserved AMP-binding sites. All ACSF2 variants were widely expressed in 11 tested tissues in geese, except that the ACSF2-2 transcript was not detected in hypothalamus, pituitary gland and granulosa cells. Subcellular localization revealed that ACSF2 is a mitochondrial matrix protein. ACSF2 mRNA level was compared between high egg production (HEP; n = 8) and low egg production (LEP; n = 10) groups and showed a lower (P < 0.05) mRNA level in the HEP group. Further experiments indicated that overexpressing ACSF2 resulted in a significant increase of caspase-3 mRNA levels and that ACSF2 knockdown triggered a decrease in the caspase-3 mRNA level in granulosa cells. Similarly, the lower caspase-3 mRNA levels were identified in ovaries of the HEP group with lower ACSF2 mRNA levels. The research showed that the ACSF2 mRNA levels had a positive correlation with caspase-3 mRNA levels in vivo (R = 0.86, P < 0.01). Our results suggest that lower ACSF2 expression promotes the laying performance of goose possibly by inhibiting granulosa cell apoptosis and facilitating follicular development.

Decreased expression of miR-874 and its tumor suppressive function in human colorectal cancer.

Dysregulation of miRNAs is associated with cancer development and progression. For example, aberrant expression of miR-874 has been found in some types of cancer. However, miR-874 expression and its clinical significance in colorectal cancer (CRC) have not yet been explored. The aim of the current study was to explore the effects of miR-874 in CRC tumorigenesis and development. Quantitative reverse-transcription PCR was performed to evaluate miR-874 levels in CRC cell lines and in 135 pairs of primary human CRC specimens and adjacent noncancerous tissues. The association of miR-874 expression with clinicopathological factors and prognosis was also analyzed. Furthermore, the effects of miR-874 on the biological behavior of CRC cells in vitro were investigated. Our results revealed that miR-874 expression was significantly downregulated in CRC cancer tissues and cell lines. Decreased miR-874 expression was significantly associated with larger tumor size, deeper invasion depth, and advanced TNM stage in vivo. Additionally, low miR-874 expression in CRC was an independent predictor of poor survival. Moreover, overexpression of miR-874 inhibited cell proliferation, invasion, and migration, and promoted cell apoptosis of the SW620 CRC cell line in vitro. Taken together, these findings indicate that miR-874 may act as a tumor suppressor in CRC, and may serve as a novel therapeutic target for miR-based therapy.

Overexpression of an endo-1,4-ß-glucanase V gene (EGV) from Trichoderma reesei leads to the accumulation of cellulase activity in transgenic rice.

The ectopic expression of cellulase in biomass can reduce the cost of biofuel conversion. This trait modification technique is highly beneficial for biofuel production. In this study, we isolated an endo-1,4-beta-glucanase gene (EGV) from Trichoderma reesei and inserted this gene downstream of a fragment encoding the signal peptide Apo-SP in a modified pCAMBIA1301 vector to obtain an Apo-SP and AsRed fusion protein. Transient expression of this fusion protein in onion epidermal cells showed that the Apo-SP signal was localized to the plastids. EGV transgenic rice plants that did not carry screening marker genes were obtained through overexpression of the pDTB double T-DNA vector. Western blotting showed that EGV was expressed in the dry straw of T0 generation transgenic rice plants and in fresh leaves of the T1 generation. More importantly, our results also showed that the peptide product of EGV in the transgenic plants folded correctly and was capable of digesting the cellulase substrate CMC. Additionally, cellulase activity remained stable in the straw that had been dried at room temperature for three months. This study presents an important technical approach for the development of transgenic rice straw that has stable cellulase activity and can be used for biofuel conversion.