Associations between cord blood metabolic factors and early-childhood growth and overweight and obesity.

Objective: This prospective cohort study was aimed at investigating the associations between cord blood metabolic factors and early-childhood growth, further elucidating the relationships between cord blood metabolites and overweight and obesity in early life. Methods: A total of 2,267 pairs of mothers and offspring were recruited in our study. Cord blood plasma was assayed for triglycerides (TGs), total cholesterol (TC), low-density lipoprotein-cholesterol (LDL-C), high-density lipoprotein-cholesterol (HDL-C), C-peptide, insulin, and glycosylated hemoglobin type A1C (HbA1c) levels. Data of anthropometric measurements were collected from offspring at birth, 6 months, 12 months, and 18 months. Multiple linear regression models were used to evaluate the correlations between cord blood metabolic factors and weight Z-scores, body mass index (BMI) Z-scores, and weight gains at the early stage of life. Forward stepwise logistic regression analyses were applied to explore the associations between cord blood metabolic factors and early-childhood overweight and obesity. Receiver operating characteristic (ROC) curve analyses were applied to determine the optimal cutoff points for cord blood metabolic factors in predicting early-childhood overweight and obesity. Results: After adjustments for covariates, cord blood TG concentrations and TG/TC ratios were negatively associated with weight Z-scores from birth to 18 months. Cord blood C-peptide and HbA1c levels were inversely associated with weight Z-scores at 6 months and 18 months. Cord blood TG concentrations and TG/TC ratios were negatively correlated with BMI Z-scores up to 18 months. Cord blood C-peptide levels and HbA1c levels were inversely correlated with BMI Z-scores at 18 months. Cord blood TG, TG/TC ratios, C-peptide, and HbA1c had negative correlations with weight gains from birth to 6 months, but the correlations attenuated as time went on. Increase in cord blood TG and HbA1c levels and TG/TC ratios were significantly associated with decreased risks of overweight and obesity at 6 months, 12 months, and 18 months. Conclusions: Cord blood metabolic factors were significantly associated with early-childhood growth patterns.

Neural pathway connectivity and discharge changes between M1 and STN in hemiparkinsonian rats.

Alterations of electrophysiological activities, such as changed spike firing rates, reshaping the firing patterns, and aberrant frequency oscillations between the subthalamic nucleus (STN) and the primary motor cortex (M1), are thought to contribute to motor impairment in Parkinson's disease (PD). However, the alterations of electrophysiological characteristics of STN and M1 in PD are still unclear, especially under specific treadmill movement. To examine the relationship between electrophysiological activity in the STN-M1 pathway, extracellular spike trains and local field potential (LFPs) of STN and M1 were simultaneously recorded during resting and movement in unilateral 6-hydroxydopamine (6-OHDA) lesioned rats. The results showed that the identified STN neurons and M1 neurons exhibited abnormal neuronal activity after dopamine loss. The dopamine depletion altered the LFP power in STN and M1 whatever in rest or movement states. Furthermore, the enhanced synchronization of LFP oscillations after dopamine loss was found in 12-35 Hz (beta frequencies) between the STN and M1 during rest and movement. In addition, STN neurons were phase-locked firing to M1 oscillations at 12-35 Hz during rest epochs in 6-OHDA lesioned rats. The dopamine depletion also impaired the anatomical connectivity between the M1 and STN by injecting anterograde neuroanatomical tracing virus into M1 in control and PD rats. Collectively, impairment of' electrophysiological activity and anatomical connectivity in the M1-STN pathway may be the basis for dysfunction of the cortico-basal ganglia circuit, correlating with motor symptoms of PD.

A machine learning-based diagnostic model for children with autism spectrum disorders complicated with intellectual disability.

Background: Early detection of children with autism spectrum disorder (ASD) and comorbid intellectual disability (ID) can help in individualized intervention. Appropriate assessment and diagnostic tools are lacking in primary care. This study aims to explore the applicability of machine learning (ML) methods in diagnosing ASD comorbid ID compared with traditional regression models. Method: From January 2017 to December 2021, 241 children with ASD, with an average age of 6.41 ± 1.96, diagnosed in the Developmental Behavior Department of the Children's Hospital Affiliated with the Medical College of Zhejiang University were included in the analysis. This study trained the traditional diagnostic models of Logistic regression (LR), Support Vector Machine (SVM), and two ensemble learning algorithms [Random Forest (RF) and XGBoost]. Socio-demographic and behavioral observation data were used to distinguish whether autistic children had combined ID. The hyperparameters adjustment uses grid search and 10-fold validation. The Boruta method is used to select variables. The model's performance was evaluated using discrimination, calibration, and decision curve analysis (DCA). Result: Among 241 autistic children, 98 (40.66%) were ASD comorbid ID. The four diagnostic models can better distinguish whether autistic children are complicated with ID, and the accuracy of SVM is the highest (0.836); SVM and XGBoost have better accuracy (0.800, 0.838); LR has the best sensitivity (0.939), followed by SVM (0.952). Regarding specificity, SVM, RF, and XGBoost performed significantly higher than LR (0.355). The AUC of ML (SVM, 0.835 [95% CI: 0.747-0.944]; RF, 0.829 [95% CI: 0.738-0.920]; XGBoost, 0.845 [95% CI: 0.734-0.937]) is not different from traditional LR (0.858 [95% CI: 0.770-0.944]). Only SVM observed a good calibration degree. Regarding DCA, LR, and SVM have higher benefits in a wider threshold range. Conclusion: Compared to the traditional regression model, ML model based on socio-demographic and behavioral observation data, especially SVM, has a better ability to distinguish whether autistic children are combined with ID.

Rapid and Visual Detection of Toxoplasma gondii in Blood Samples from Pet Cats and Dogs by Loop-Mediated Isothermal Amplification.

Background: Toxoplasma gondii is an obligate intracellular parasite that invades nearly all nucleated cells of a broad spectrum of vertebrate hosts, and which may cause serious disease in immunocompromised patients, as well as in the immunologically incompetent fetus. This study aimed to establish a loop-mediated isothermal amplification (LAMP) technique to rapidly detect T. gondii in the blood infection by targeting the 529 bp repeat element of T. gondii. Materials and Methods: A turbidity monitoring system, together with visual reagent, was used to test the amplification result of the LAMP assay. In addition, the specificity and sensitivity of the LAMP assay were measured. Results: The results suggest that the successfully established LAMP assay profile can detect the DNA of T. gondii at 67°C within 40 min. The limit of detection of the LAMP assay was 101 copies/µL. No cross reaction occurred with Plasmodium vivax, Toxocara cati, Clonorchis sinensi, Spirometra mansoni or Cryptosporidium parvum. We validated the developed LAMP assay by detecting T. gondii in DNA extracted from 353 blood samples collected from domestic cats and dogs. The percentages of positive results in detecting these blood samples by LAMP and conventional PCR were 5.38% and 2.83%, respectively. Conclusions: Our findings show that the developed LAMP assay offers higher analytical sensitivity than conventional PCR and good analytical specificity, minimizes aerosol contamination, and can be applied to on-site rapid detection of T. gondii.

[Characteristics of Xin'an medical masters in treatment of bi syndrome with acupuncture and moxibustion based on consolidating the root and cultivating the primary].

Based on the theory of consolidating the root and cultivating the primary, the paper collates and reviews the theoretical evidences and the characteristics of Xin'an medical masters in treatment of bi syndrome with acupuncture and moxibustion so as to provide the ideas for further research. Xin'an medical masters thoroughly acquainted with the theory of consolidating the root and cultivating the primary in treatment of bi syndrome with acupuncture and moxibustion, emphasizing the regulation of qi and blood, yin and yang, the nutrient qi and the defensive qi; and replenishing the middle jiao (spleen and stomach) and the lower jiao (kidney). The acupoint selection was distinctive, in which, the acupoints were selected and stimulated in terms of the etiology and the pathogenesis of diseases, as well as the properties of special points. The remarkable therapeutic effect on bi syndrome was ensured through specially selecting he-sea points, ashi points and "yin-yang opposite" points; effectively using the penetrating needling technique and moxibustion method and combining acupuncture with herbal medication.

Intelligence profiles and adaptive behaviors of high-functioning autism spectrum disorder and developmental speech and language disorders.

Objective: The present study was aimed at investigating the intelligence profiles and adaptive behaviors of children with high-functioning autism spectrum disorder (HFASD) and developmental speech and language disorders (DSLDs). We compared the similarities and differences of cognitive capabilities and adaptive functions and explored their correlations in the HFASD and DSLDs groups. Methods: 128 patients with HFASD, 111 patients with DSLDs and 114 typically developing (TD) children were enrolled into our study. Wechsler Intelligence Scale for Children-IV (WISC-IV) and Adaptive Behavior Assessment System-II (ABAS-II) were respectively applied to evaluate intelligence profiles and adaptive behaviors. Intelligence quotient (IQ) scores and adaptive functioning scores among the HFASD, DSLDs and TD groups were compared through one-way ANOVA. Pearson correlation coefficient was applied to examine the relationships between WISC indices and ABAS domains. Results: Outcomes showed significantly poorer intelligence profiles and adaptive behaviors in HFASD and DSLDs groups. Both children with HFASD and DSLDs demonstrated impairments in verbal comprehension and executive functions. Processing speed and working memory were the predominant defects of children with HFASD and DSLDs in the field of executive functions, respectively. Whereas perceptual reasoning was a relative strength for them. Children with DSLDs had balanced scores of all the domains in ABAS-II; nevertheless, HFASD individuals demonstrated striking impairments in Social domain. Correlation analysis showed IQs of children with HFASD were positively correlated with all the domains and General Adaptive Composite (GAC) of ABAS-II. Additionally, IQs were positively correlated with Conceptual domain and GAC for children with DSLDs. Compared with DSLDs group, intelligence displayed stronger correlations with adaptive behaviors in HFASD group. Conclusion: Our study expanded insights regarding intelligence profiles and adaptive behaviors of children with HFASD and DSLDs. Moreover, this study made breakthroughs in discovering positive correlations between IQs and adaptive functions in the two neurodevelopmental disorders.

Absolute Quantification of Allergen Glb33 in Rice by Liquid Chromatography-Mass Spectrometry using Two Isotope-Labeled Standard Peptides.

Allergen Glb33 is an important allergen in rice that can cause allergic reactions such as asthma and atopic dermatitis. However, knowledge of the content in rice is sparse. In the present work, an absolute protein quantification method was established for allergen Glb33 in rice samples using liquid chromatography-tandem mass spectrometry. After extraction of allergen Glb33 from rice grains using salt solution, the isotope-labeled peptide internal standard was added to the extract, followed by enzymatic digestion with trypsin. The signature peptide and its isotope-labeled analogue from the tryptic hydrolysates of allergen Glb33 and the internal standard were detected by liquid chromatography-tandem mass spectrometry. The quantitative bias caused by tryptic efficiency and matrix effect was corrected by using two isotope-labeled standard peptides. The method exhibited good linearity in the range of 1-200 nM, with coefficients of determination of R2 > 0.998. A high sensitivity was observed, with a limit of quantification of 0.97 nM. Mean recoveries obtained from different rice matrices ranged from 82.7%-98.1% with precision <8.5% in intraday trials ( n = 6), while mean recoveries were from 75.1%-107.4% with precision <14.6% in interday trials ( n = 14). The developed method was successfully applied to the analysis of allergen Glb33 in 24 different rice cultivars.

Evaluation of Whole Blood Trace Element Levels in Chinese Children with Autism Spectrum Disorder.

Autism spectrum disorder (ASD) is a neurodevelopmental disorder, which has increased markedly during the last decades. Essential trace elements play an important role in neurological function and their imbalances are common in children with ASD. The objective of the present study was to investigate whole blood levels of trace elements including zinc (Zn), copper (Cu), iron (Fe), and magnesium (Mg) in Chinese children with ASD. In total, 113 children diagnosed with ASD and 141 age-matched and gender-matched neurotypical children, divided into two gender and age groups of preschool age (2-5 years old) and school (6-10 years old) age, were examined. The quantitative analyses of whole blood trace element contents were performed by using flame atomic absorption spectroscopy. In the present study, the children with ASD generally had lower whole blood levels of Zn than the neurotypical controls. No significant differences in the whole blood Cu, Zn/Cu ratio, Fe, or Mg was detected between the ASD group and the control group. It is notable that whole blood Fe level in boys with ASD was significantly higher than in girls with ASD, and was nearly significant when compared with the control level of boys. After stratification for age, a significant 6% decrease in whole blood Zn levels was detected in preschool-aged children with ASD as compared to the control values. However, this significant ASD-related change was not detected in school-aged children. The whole blood Zn level and Zn/Cu ratio were significantly increased in school-aged children than in preschool-aged children in both ASD and control group. In addition, school-aged children with ASD had a significantly higher level of whole blood Fe than preschool-aged children with ASD. The results of the present study suggest an association between whole blood levels of Zn in Chinese children with ASD.

The Application of the First Year Inventory for ASD Screening in China.

PURPOSE: The First Year Inventory (FYI) is a parent-report instrument, and is developed to assess behaviors of 12-month-old infants that could suggest risk for an eventual diagnosis of autism. This study was designed to examine the application of the FYI in the Chinese community. DESIGN AND METHODS: FYIs were completed at a community health center by 541 families during the child's physical examination at 12 months of age from 2013 to 2015. The weighted risk scores used in this study were based on US norms, and compared the FYI differences between China and the U.S. RESULTS: The total risk scores ranged from 5 to 42 points; the 95th percentile cutoff was 27.00(9.8 points higher than the 95th percentile cutoff in the US), the 98th percentile cutoff was 29.66(7.04 points higher than the 98th percentile cutoff in the US), and the 99th percentile cutoff was 31.83. Higher risk scores were found for boys than girls. Mothers with a junior college education reported significantly higher FYI risk scores than other three groups including high school, college graduates and post-graduates. CONCLUSIONS: There were no significant effects of birth parity, investigator, or investigation year on risk scores. Large-scale longitudinal research is encouraged in the future to develop an early detection model of autism in China.

Differentially expressed circular RNAs in systemic lupus erythematosus and their clinical significance.

It has been reported that circRNAs are diff ;erentially expressed in many diseases and can be used as new biomarker to facilitate disease diagnosis. Circular RNAs (circRNAs) microarray were used to identify dysregulated circRNAs in plasma of systemic lupus erythematosus (SLE) patients. Then, we confirmed the microarray data by quantitative reverse transcription polymerase chain reaction (qRT-PCR) in both plasma and peripheral blood mononuclear cells (PBMCs) of SLE. One hundred and twelve circRNAs were identified to dysregulated expressed in plasma of SLE as compared to healthy controls. The results of qRT-PCR showed that the levels of hsa_circRNA_407176 and hsa_circRNA_001308 were decreased in both plasma and PBMCs of SLE when compared with healthy controls. The receiver operating characteristic (ROC) curve area of hsa_circRNA_407176 and hsa_circRNA_001308 in plasma were 0.599 and 0.662, respectively. The area under the ROC curves of hsa_circRNA_407176, hsa_circRNA_406567 and hsa_circRNA_001308 in PBMCs were 0.806, 0.744, and 0.722, respectively. Our study illustrated that hsa_circRNA_407176 and hsa_circRNA_001308 in plasma and PBMCs could be potential biomarkers for SLE.

Effects of pharmaceuticals on microbial communities and activity of soil enzymes in mesocosm-scale constructed wetlands.

Cyperus alternifolius based mesocosm-scale constructed wetland was employed to remove pharmaceuticals. We investigated the microbial community composition using phosphor lipid fatty acids (PFLAs) analysis and substrate enzyme activity during long-term exposure to pharmaceuticals in mesocosm-scale constructed wetlands. The results showed that there was no visible inhibition effect of pharmaceuticals on CW substrate enzymes activities in the experimental range (0-500 µg/L). Microbial communities, as revealed by PFLAs, were enhanced by the presence of plants, while the PFLAs content was highest when the pharmaceutical concentration was 10 µg/L or 30 µg/L at CWs. Except for anaerobic bacteria and Saturated fatty acids, the maximum PLFAs levels were reached when the pharmaceuticals were 10 µg/L or 30 µg/L, while Bacteria, G (-), fungal bacteria, Aerobic bacteria and Monounsaturated fatty acids were remarkably affected by high pharmaceuticals (100-500 µg/L). However, the main microbial florae were not changed among the treatments. In this study, the removal efficiencies of the studied pharmaceuticals in Planted (30) was greatest, which could be attributed to the higher microbial biomass. These results indicate that C. alternifolius can phytoremediate pharmaceutical-contaminated waters in CWs. Individual fatty acid cannot be used to represent specific species; therefore, more approaches to species identification such as rRNA-based methods must be included in future studies to better understand the metabolic mechanisms of microorganisms involved in the removal of studied pharmaceuticals and improve the performance of CWs.

Circular RNA expression profile and potential function of hsa_circ_0045272 in systemic lupus erythematosus.

Circular RNAs (circRNAs) represent a class of non-coding RNAs that form covalently closed RNA circles with extensive expression and conservation in mammals. Circular RNAs regulate gene expression through acting as competitive endogenous RNAs (ceRNAs) and modulating gene transcription. Accumulating evidence supports the implication of circRNAs in a variety of human diseases, but studies of circRNA role in systemic lupus erythematosus (SLE) are lacking. The present study measured the circRNA expression profiles in T cells from patients with SLE and healthy controls with human circRNA microarray and identified 127 differentially expressed circRNAs in SLE patients. Down-regulation of hsa_circ_0045272 in SLE T cells was verified with quantitative PCR. Jurkat cells with stable hsa_circ_0045272 knockdown were generated using specific lentiviral short hairpin RNA for functional studies. Flow cytometric analysis indicated that hsa_circ_0045272 knockdown significantly up-regulated the early apoptosis of Jurkat cells. Meanwhile, ELISA showed that hsa_circ_0045272 knockdown significantly enhanced interleukin-2 production of activated Jurkat cells. Then, ceRNAs were predicted for hsa_circ_0045272 and the significant down-regulation of two mRNAs predicted as ceRNAs, NM_003466 (PAX8) and NM_015177 (DTX4), but not their corresponding proteins, was validated. Furthermore, dual luciferase reporter assay indicated binding of hsa_circ_0045272 with hsa-miR-6127. Circular RNA-mRNA co-expression networks showed the correlation of circRNAs with mRNAs and provided additional clues to circRNA functions. Our study demonstrated dysregulated circRNAs in SLE and revealed the function of hsa_circ_0045272 in negatively regulating apoptosis and interleukin-2 secretion and its potential mechanism. The implication of hsa_circ_0045272 and other abnormal circRNAs in SLE merits further investigation.

Sulfur supply reduces cadmium uptake and translocation in rice grains (Oryza sativa L.) by enhancing iron plaque formation, cadmium chelation and vacuolar sequestration.

Sulfur (S) fertilizer application in rice (Oryza sativa L.) is crucial in determining rice grain productivity and quality. However, little information is available concerning the effect of S supply on cadmium (Cd) uptake and translocation in rice. In this study, both hydroponic and soil experiments were conducted to investigate the influence of S supply on Cd accumulation in rice under two Cd levels (0 and 50 µM), combined with three S concentrations (0, 2.64 and 5.28 mM). The moderate and excessive S supply (2.64 and 5.28 mM) tended to increase plant growth, root length, root and shoot dry weights of rice seedlings, and significantly decreased Cd concentrations in rice plants and grains in the absence or presence of Cd. The subcellular distribution and chemical forms of Cd in roots and shoots also varied with S supply levels. The decreased Cd uptake and translocation in rice grains could be ascribed to the enhanced formation of iron (Fe) plaque on the root surfaces and increased Cd chelation and vacuolar sequestration in roots, since Fe, Mn concentrations in Fe plaque, glutathione and phytochelatins contents, as well as phytochelatin synthase (OsPCS) and tonoplast heavy metal ATPase (OsHMA3) expressions in roots significantly increased with increased S supply. This work provides more insight into the mechanisms of Cd uptake and translocation in rice, and will be helpful for developing strategies to reduce rice grain Cd through S fertilizer application in Cd-contaminated soil.

Xiaochaihutang Inhibits the Activation of Hepatic Stellate Cell Line T6 Through the Nrf2 Pathway.

Xiaochaihutang (XCHT) is one of classic prescriptions in Treatise on Febrile Diseases in China which was reported to have the effect of anti-hepatic fibrosis in vivo. Activation of hepatic stellate cells (HSCs) is now well established as a central driver of fibrosis in liver injury. Nuclear factor erythroid 2-related factor 2 (Nrf2) is an important element for anti-oxidative damage which is one of the key factors responsible for occurrence. This study was to investigate the effect of XCHT compound serum on HSCs activation and focus on the Nrf2 pathway. Rats in treatment groups were given the appropriate doses of XCHT granules (5 g/kg) and Silybin (50 mg/kg) for 6 days, and the serum were obtained. The compound serum was used to intervene HSCs. The results found that XCHT compound serum significantly inhibited the proliferation of HSCT6 cells. The number of α-SMA positive stained cells in HSCT6 cells and the content of Collagen type I (collagen-I) in supernatant were significantly decreased indicating suppression of activated HSCs. Compared with the control group, the nuclear transcription of Nrf2 and the expressions of Nqo1, GCLC, and GCLM were significantly increased in XCHT group. However, the effects of XCHT were inhibited in Nrf2-siRNA transfected HSCT6 cells. These studies demonstrated that XCHT could inhibit HSCT6 cell proliferation and activation. The mechanism might be related to up-regulation of the Nrf2 pathway against oxidative stress.

Jun dimerization protein 2 controls hypoxia-induced replicative senescence via both the p16Ink4a-pRb and Arf-p53 pathways.

The main regulators of replicative senescence in mice are p16Ink4a and Arf, inhibitors of cell cycle progression. Jun dimerization protein 2 (JDP2)-deficient mouse embryonic fibroblasts are resistant to replicative senescence through recruitment of the Polycomb repressive complexes 1 and 2 to the promoter of the gene that encodes p16Ink4a and inhibits the methylation of lysine 27 of the histone H3 locus. However, whether or not JDP2 is able to regulate the chromatin signaling of either p16Ink4a-pRb or Arf-p53, or both, in response to oxidative stress remains elusive. Thus, this study sought to clarify this point. We demonstrated that the introduction of JDP2 leads to upregulation of p16Ink4a and Arf and decreases cell proliferation in the presence of environmental (20% O2), but not in low (3% O2) oxygen. JDP2-mediated growth suppression was inhibited by the downregulation of both p16Ink4a and Arf. Conversely, the forced expression of p16Ink4a or Arf inhibited cell growth even in the absence of JDP2. The downregulation of both the p53 and pRb pathways, but not each individually, was sufficient to block JDP2-dependent growth inhibition. These data suggest that JDP2 induces p16Ink4a and Arf by mediating signals from oxidative stress, resulting in cell cycle arrest via both the p16Ink4a-pRb and Arf-p53 pathways.

Direct Determination of Six Cytokinin Nucleotide Monophosphates in Coconut Flesh by Reversed-Phase Liquid Chromatography-Tandem Mass Spectrometry.

Coconut contains many uncharacterized cytokinins that have important physiological effects in plants and humans. In this work, a method based on liquid chromatography-tandem mass spectrometry was developed for identification and quantification of six cytokinin nucleotide monophosphates in coconut flesh. Excellent separation was achieved using a low-coverage C18 bonded-phase column with an acidic mobile phase, which greatly improved the retention of target compounds. To enable high-throughput analysis, a single-step solid-phase extraction using mixed-mode anion-exchange cartridges was employed for sample preparation. This proved to be an effective method to minimize matrix effects and ensure high selectivity. The limits of detection varied from 0.06 to 0.3 ng/mL, and the limits of quantification ranged from 0.2 to 1.0 ng/mL. The linearity was statistically verified over 2 orders of magnitude, giving a coefficient of determination (R2) greater than 0.9981. The mean recoveries were from 81 to 108%; the intraday precision (n = 6) was less than 11%; and the interday precision (n = 11) was within 14%. The developed method was applied to the determination of cytokinin nucleotide monophosphates in coconut flesh samples, and four of them were successfully identified and quantified. The results showed that trans-zeatin riboside-5'-monophosphate was the dominant cytokinin, with a concentration of 2.7-34.2 ng/g, followed by N6-isopentenyladenosine-5'-monophosphate (≤12.9 ng/g), while the concentrations of cis-zeatin riboside-5'-monophosphate and dihydrozeatin riboside-5'-monophosphate were less than 2.2 and 4.9 ng/g, respectively.

Insights into the molecular mechanism of the responses for Cyperus alternifolius to PhACs stress in constructed wetlands.

Cyperus alternifolius has been widely reported to be an effective phytoremediation plant in constructed wetland systems (CWs). In this context, an integrated biochemical and proteomic analysis of C. alternifolius leaves exposed to pharmaceutically active compounds (PhACs) in CWs was conducted to understand the mechanism of phytoremediation. The obtained results showed the antioxidant enzyme activities were induced throughout the experiment; however over time, the malondialdehyde content is not significantly different from the control and the photosynthetic pigment contents in plant were subsequently slowly recovered. Therefore, we concluded that reactive oxygen species could be effectively counteracted by the enhanced antioxidant enzyme activities, and therefore the photosynthetic pigments were ultimately restored. Leaf extract proteome maps were obtained through 2-DE, and an average of 55, 49, and 24 spots were significantly altered by 30, 100, and 500 µg/L of PhACs over the control, respectively. Protein expression patterns showed that proteins in C. alternifolius leaves are associated with photosynthesis, energy metabolism, defense, and protein synthesis. Moreover, the most relevant pathways modulated by PhACs were photosynthesis and energy metabolism. The protein expression involved in antioxidant defense and stress response generally increased in all the PhAC treatments. The regulated proteins may favor PhAC degradation in CWs; however, the role of these proteins in degrading PhACs remains unknown; further biochemical studies should be conducted. This study indicated that C. alternifolius can tolerate multiple PhACs.

Production of germline transgenic pigs co-expressing double fluorescent proteins by lentiviral vector.

Genomic integration of transgene by lentiviral vector has been proved an efficient method to produce single-transgenic animals. But it failed to create multi-gene transgenic offspring. Here, we have exploited lentivirus to generate the double-transgenic piglets through the female germline. The recombinant lentivirus containing fluorescent proteins genes (DsRed1 and Venus) were injected into the perivitelline space of 2-cell stage in vitro porcine embryos. Compared to control group, there was no significantly decreased in the proportion of blastocysts, and the two fluorescent protein genes were co-expressed in almost all the injected embryos. Total of 32 injected in vitro embryos were transferred to 2 recipients. One recipient gave birth of three live offspring, and one female piglet was identified as genomic transgene integration by PCR analysis. Subsequently, the female transgenic founder was mated naturally with a wild-type boar and gave birth of two litters of total 23 F(1) generation piglets, among which Venus and DsRed1 genes were detected in 11 piglets and 10 kinds of organs by PCR and RT-PCR respectively. The co-expression of two fluorescent proteins was visible in four different frozen tissue sections from the RT-PCR positive piglets, and 3 to 5 copies of the transgenes were detected to be integrated into the second generation genome by southern blotting analysis. The transgenes were heritable and stably integrated in the F(1) generation. The results indicated for the first time that lentiviral vector combined with natural mating has the potential to become a simple and practical technology to create germline double-transgenic livestock or biomedical animals.

Independent and Combined Effects of Maternal Prepregnancy Body Mass Index and Gestational Weight Gain on Offspring Growth at 0-3 Years of Age.

Background. The objective of this study was to investigate the independent and combined effects of maternal prepregnancy body mass index (BMI) and gestational weight gain (GWG) on offspring growth at 0-3 years old. Methods. A total of 826 pairs of nondiabetic mothers and their offspring were recruited in this study. Maternal information was abstracted from medical records and questionnaires. Offspring growth trajectories of weights and BMIs were depicted based on anthropometric measurements. Results. Offspring of mothers who were prepregnancy overweight/obese or obtained excessive GWGs continuously had greater weight and BMI Z-scores throughout the first 3 years of life. Children of prepregnancy overweight/obese mothers with excessive GWGs had a phenotype of higher weight and BMI Z-scores than those prepregnancy overweight/obese ones with nonexcessive GWGs from birth to 18 months. Maternal excessive GWGs increased offspring's risk of overweight/obesity at 12 months (AOR = 1.43, 95% CI: 1.03-2.00) and 24 months (AOR = 1.51, 95% CI: 1.02-2.25). Combination of excessive prepregnancy BMIs and GWGs was significantly associated with offspring's overweight/obesity at 30 months (AOR = 2.98, 95% CI: 1.36-6.53). Conclusions. Maternal prepregnancy overweight/obesity and excessive GWG are both significantly associated with rapid offspring growth from birth to 3 years old. Excessive GWGs strengthen the effects of high maternal prepregnancy BMIs on excessive offspring growth during their early life.