Drought responses and population differentiation of Calohypnum plumiforme inferred from comparative transcriptome analysis.

Bryophytes, known as poikilohydric plants, possess vegetative desiccation-tolerant (DT) ability to withstand water deficit stress. Consequently, they offer valuable genetic resources for enhancing resistance to water scarcity stress. In this research, we examined the physiological, phytohormonal, and transcriptomic changes in DT mosses Calohypnum plumiforme from two populations, with and without desiccation treatment. Comparative analysis revealed population differentiation at physiological, gene sequence, and expression levels. Under desiccation stress, the activities of superoxide dismutase (SOD) and peroxidase (POD) showed significant increases, along with elevation of soluble sugars and proteins, consistent with the transcriptome changes. Notable activation of the bypass pathway of JA biosynthesis suggested their roles in compensating for JA accumulation. Furthermore, our analysis revealed significant correlations among phytohormones and DEGs in their respective signaling pathway, indicating potential complex interplays of hormones in C plumiforme. Protein phosphatase 2C (PP2C) in the abscisic acid signaling pathway emerged as the pivotal hub in the phytohormone crosstalk regulation network. Overall, this study was one of the first comprehensive transcriptome analyses of moss C. plumiforme under slow desiccation rates, expanding our knowledge of bryophyte transcriptomes and shedding light on the gene regulatory network involved in response to desiccation, as well as the evolutionary processes of local adaptation across moss populations.

Genotoxic mode of action and threshold exploration of 2-methyl furan under 120-day sub-chronic exposure in male Sprague-Dawley rats.

2-Methylfuran (2-MF) is an important member of the furan family generated during food thermal processing. An in-vivo multiple endpoint genotoxicity assessment system was applied to explore the genotoxic mode of action and threshold of 2-MF. Male Sprague-Dawley rats received 2-MF by oral gavage at doses of 0.16, 0.625, 2.5, and 10 mg/kg.bw/day for 120 days. An additional 15 days were granted for recovery. The Pig-a gene mutation frequency of RET and RBC showed significant increases among the 2-MF groups on day 120. After a 15-day recovery period, the Pig-a gene mutation frequency returned to levels similar to those in the vehicle control. The tail intensity (TI) values of peripheral blood cells at a dose of 10 mg/kg.bw/day significantly increased from day 4 and remained at a high level after the recovery period. No statistical difference was found in the micronucleus frequency of peripheral blood between any 2-MF dose group and the corn oil group at any timepoint. 2-MF may not induce the production of micronuclei, but it could cause DNA breakage. It could not be ruled out that 2-MF may accumulate in vivo and cause gene mutations. Hence, DNA, other than the spindle, may be directly targeted. The mode of action of 2-MF may be that it was metabolized by EPHX1 to more DNA-active metabolites, thus leading to oxidative and direct DNA damage. The point of departure (PoD) of 2-MF-induced genotoxicity was derived as 0.506 mg/kg bw/day.

A white-light-emitting sensor array based on cucurbit[8]uril for quantitative detection of multicomponent nitroaniline isomers.

A white-light-emitting supramolecular complex through supramolecular interactions has been assembled; the white luminescent supramolecular complex exhibits two emission spectra. Based on this, a dual-channel white-light array sensor was constructed. The results show that it can quickly identify and detect nitroaniline isomer pollutants (p-nitroaniline, m-nitroaniline, o-nitroaniline). When these three nitroaniline isomers were added to the supramolecular white-light array sensor, the fluorescence intensity of the white-light complex decreased to varying degrees. Linear discriminant analysis (LDA) showed that the supramolecular white-light array sensor could recognize and distinguish three nitroaniline isomers and could classify mixtures containing different concentrations. Factor 1 of the array had a good linear relationship with the concentration of pollutants, and the detection limit (LOD) was as low as 0.7 µM. The method has good reproducibility and stability. In addition, it can also qualitatively detect the nitroaniline isomers in river water and contaminated rice seedling extract. It provides an ideal platform for constructing multiresponse sensors.

Supramolecular fluorescent probe based on a host-guest complex for amino acids recognition and detection in aqueous solution.

A supramolecular fluorescent probe based on a host-guest complex has been developed for amino acid recognition and detection in aqueous solution. Cucurbit[7]uril (Q[7]) with 4-(4-dimethylamino-styrene) quinoline (DSQ) formed a fluorescent probe (DSQ@Q[7]). The DSQ@Q[7] fluorescent probe nearly generated changes in fluorescence in response to four amino acids (arginine, histidine, phenylalanine and tryptophan). These changes were attributed to the host-guest interaction between DSQ@Q[7] and amino acids, which occurred as a consequence of the subtle cooperation of ionic dipole and hydrogen bonding. Linear discriminant analysis showed that the fluorescent probe could recognize and distinguish four amino acids, and a mixture with different concentration ratios could be well categorized in ultrapure water and tap water.

Supramolecular Fluorescence Sensor Array Based on Cucurbit[8]uril Complexes Used for the Detection of Multiplex Quaternary Ammonium Pesticides.

The simultaneous detection of multiple quaternary ammonium pesticides (QAPs) in water is a challenge due to their high solubility in water and similar structures. In this paper, we have developed a quadruple-channel supramolecular fluorescence sensor array for the simultaneous analysis of five QAPs, including paraquat (PQ), diquat (DQ), difenzoquat (DFQ), mepiquat (MQ), and chlormequat (CQ). Not only were QAP samples of different concentrations (10, 50, and 300 µM) in water distinguished with 100% accuracy but also single QAP and binary QAP mixed samples (DFQ-DQ) were sensitively quantified. Our experimental interference study confirmed that the developed array has good anti-interference ability. The array can quickly identify five QAPs in river and tap water samples. In addition, it also qualitatively detected QAP residues in Chinese cabbage and wheat seedlings extract. This array has rich output signals, low cost, easy preparation, and simple technology, demonstrating great potential in environmental analysis.

A supramolecular fluorescence array sensor for toxic heavy metal ion detection in environmental water and rice seedling extracts.

A supramolecular fluorescence array sensor based on cucurbituril-dye host-guest complexes (6-QAA@Q[7], PyY@Q[7], and TO@Q[8]) was constructed. The results showed that it can quickly identify and detect toxic heavy metal ions, such as Ag+, Cr3+, Hg2+, Ni2+, and Pb2+. When these five toxic heavy metal ions were added to the supramolecular fluorescence array sensor, different fluorescence responses were produced due to the different binding capacities of the metal ions to the cucurbituril-dye complex. Linear discriminant analysis (LDA) showed that the supramolecular fluorescence array sensor could identify and distinguish these five toxic heavy metal ions and a mixture containing different concentration ratios could be classified. The linear correlation between the metal ion concentration and factor 1 (F1) was strong, and the detection limit (LOD) was as low as 10-6-10-7 mol L-1. These five toxic heavy metal ions in environmental water and rice seedling extracts were identified using the supramolecular fluorescence array sensor. This sensor provides a quick and convenient method for monitoring toxic heavy metal ions in sewage.

Reclaiming Family, Reimaging Queer Relationality.

Family is an important issue in imagining queer modes of existence. In this essay, I argue for a queer relationality that is structured around biogenetic family, a site that is often marginalized and negated in dominant Euro-American queer discourse. Informed by queer of color critique and postcolonial feminism, this essay affirms the relational framework in understanding the everyday struggles of queer subjects. Situated in the context of Chinese society, I investigate a queer relationality that centers ambivalence and inbetweenness as queer modes of positioning, challenging the teleological narrative of queerness that is characterized by oppositionality, singularity, and anti-relationality.

A colorimetric supramolecular sensor array based on charge-transfer complexes for multiplex aniline and phenolic pollutants detection.

In order to effectively monitor toxic and harmful substances in sewage discharge, a rapid, highly sensitive detection of complex pollutants with similar structures has become an urgent problem to be solved. In this paper, a supramolecular colorimetric array sensor based on charge-transfer complex was constructed, which can quickly detect aniline and phenol pollutants (such as p-phenylenediamine, m-phenylenediamine, o-phenylenediamine, m-aminophenol, hydroquinone, and resorcinol) with similar structures. When six anilines and phenol isomers with similar structures were added to the supramolecular colorimetric array sensor, different color changes were produced under natural light. Linear discriminant analysis (LDA) showed that the supramolecular colorimetric array sensor could recognize and distinguish these isomers, and a mixture with different concentration ratios could be well categorized. The total Euclidean distance (TED) of an array with pollutant concentrations had a good linear correlation, and the detection limit (LOD) was as low as 10-5-10-6 mol L-1. Six anilines and phenol isomers in real samples were identified by supramolecular colorimetric array sensor. 1H NMR results showed that the formation of charge transfer complexes in Q[8] cavity may be the cause of color change. This work provides a fast and convenient experimental basis for monitoring the complex structure pollutants in sewage discharge.

Lab-on-a-Molecule Probe: Multitarget Detection of Five Aromatic Pesticides Using a Supramolecular Probe under Single Wavelength Excitation.

In order to prevent and control the effects of pesticide residues on human health and the ecological environment, the rapid, highly sensitive, and selective detection of multiple pesticide residues has become an urgent problem to be solved. Herein, a lab-on-a-molecule probe based on a host-guest complex (ThT@Q[8] probe) has been developed to simultaneously analyze multiple aromatic pesticides under single wavelength excitation, such as fuberidazole, thiabendazole, carbendazim, thidiazuron, and tricyclazole. The fluorescence titration spectra of the ThT@Q[8] probe with the five pesticides mentioned above showed that the fluorescence intensity exhibited a good linear correlation with the pesticide concentration and the limit of detection was as low as 10-7 M. Because the ThT@Q[8] probe exhibits diverse fluorescence color changes to the five pesticides studied under a 365 nm ultraviolet lamp, we fabricated a single probe used to detect multiple analytes in the RGB triple channel by extracting the RGB variations. Principal component analysis and linear discriminant analysis proved that the ThT@Q[8] probe can recognize and distinguish five pesticides and can be applied at different concentrations. In real samples, the ThT@Q[8] probe recognized and distinguished five pesticides in tap water and Huaxi River water. The 1H NMR spectra results proved that a charge-transfer complex of ThT and pesticides in the Q[8] cavity may be formed. Moreover, we selected a test strip as a carrier to detect pesticides. The results indicate it can be used to quickly and conveniently detect different pesticides due to the rapid color change. Besides, the ThT@Q[8] probe has good cell permeability and can be used to detect pesticide residues in living cells. This work has laid the foundation for the qualitative and quantitative multitarget detection of pesticide residues.

[Associations between birth outcomes with blood pressure level and risk of high blood pressure in childhood].

OBJECTIVE: To explore the associations between birth outcomes and blood pressure, and study the sex dimorphism of these associations. METHODS: With a multistage cluster random sampling method, 62 168 children were recruited in seven provinces of China in September of 2013, with 32 064 boys and 30 104 girls, median age of 10.74 years and mean birth weight of 3.3 kg, 49 843 single birth(97%), 1339 twin(2.6%), 180 triplet or more(0.4%). Questionnaire investigation and physical examination were conducted in the present study. Weight, height and blood pressure were measured in the physical examination. Demographic characteristics, birth outcomes(including birth weight and number of births), dietary behavior, physical activities were measured by questionnaire. The widely used age-, gender-and height-specific high blood pressure standard developed by American CDC was used for the present study. Multivariate linear and logistic regression analysis were conducted to study the associations between birth outcomes and blood pressure level or high blood pressure(HBP), and also sex dimorphism of these associations was explored. RESULTS: A total of 5933 children were categorized as having high blood pressure in the 62 168 participants(9.5%). With stratified analyses by birth weight category, only in the low-birth-weight strata birth weight was significantly inversely associated with systolic blood pressure(SBP) and diastolic blood pressure(DBP) with potential covariates adjusted(SBP: b=-1.628, 95%CI-2.571--0.685, P=0.001; DBP: b=-1.463, 95%CI-2.186--0.740, P<0.001). While compared with the non-low birth weight children, low birth weight was not associated with higher risk of HBP(P>0.05). Compared with those boys born as singleton, boys born as one of the twins have a 36.4% higher risk of HBP(OR=1.364, 95%CI 1.049-1.774), while in girls no such significant association was found. Additionally, compared with a term birth, overdue birth and preterm birth was not associated with higher risk of HBP(P>0.05). CONCLUSION: Birth weight and singleton or not were associated with childhood blood pressure levels and higher risk of high blood pressure, and some associations were sex specific. Gender differences should be paid attention to in the prevention and control of high blood pressure in children and adolescents in the future, and the prevention and control should be focused on low-birth weight children or twin boys.

Prediction of Liver Weight Recovery by an Integrated Metabolomics and Machine Learning Approach After 2/3 Partial Hepatectomy.

Liver has an ability to regenerate itself in mammals, whereas the mechanism has not been fully explained. Here we used a GC/MS-based metabolomic method to profile the dynamic endogenous metabolic change in the serum of C57BL/6J mice at different times after 2/3 partial hepatectomy (PHx), and nine machine learning methods including Least Absolute Shrinkage and Selection Operator Regression (LASSO), Partial Least Squares Regression (PLS), Principal Components Regression (PCR), k-Nearest Neighbors (KNN), Support Vector Machines (SVM), Random Forest (RF), eXtreme Gradient Boosting (xgbDART), Neural Network (NNET) and Bayesian Regularized Neural Network (BRNN) were used for regression between the liver index and metabolomic data at different stages of liver regeneration. We found a tree-based random forest method that had the minimum average Mean Absolute Error (MAE), Root Mean Squared Error (RMSE) and the maximum R square (R2) and is time-saving. Furthermore, variable of importance in the project (VIP) analysis of RF method was performed and metabolites with VIP ranked top 20 were selected as the most critical metabolites contributing to the model. Ornithine, phenylalanine, 2-hydroxybutyric acid, lysine, etc. were chosen as the most important metabolites which had strong correlations with the liver index. Further pathway analysis found Arginine biosynthesis, Pantothenate and CoA biosynthesis, Galactose metabolism, Valine, leucine and isoleucine degradation were the most influenced pathways. In summary, several amino acid metabolic pathways and glucose metabolism pathway were dynamically changed during liver regeneration. The RF method showed advantages for predicting the liver index after PHx over other machine learning methods used and a metabolic clock containing four metabolites is established to predict the liver index during liver regeneration.

Effect of high-quality nursing on psychological status and prognosis of patients undergoing brain tumor surgery.

OBJECTIVE: To investigate the effect of high-quality nursing on psychological status and prognosis of patients undergoing brain tumor surgery. METHODS: One hundred and ten patients undergoing brain tumor surgery were divided into control group (n=55, receiving routine nursing) and observation group (n=55, receiving high-quality nursing). The psychological status (Hamilton Anxiety Rating Scale (HAMA) and Hamilton Depression Rating Scale (HAMD)), quality of life (Generic Quality of Life Inventory-74 Scale (GQOLI-74)), prognosis (Glasgow Outcome Scale (GOS)), self-efficacy (Strategies Used by People to Promote Health Scale (SUPPH)) and complications of patients in both groups were recorded and compared. RESULTS: After intervention, scores of HAMA and HAMD in both groups decreased, while scores of GOS, GQOLI-74 and SUPPH in both groups increased (all P<0.05). Compared with the control group, scores of HAMA and HAMD in the observation group were lower after intervention, while scores of GOS, GQOLI-74 and SUPPH were higher (all P<0.05). There was no significant difference in mortality between the two groups (P>0.05). The overall complication rate in the observation group was lower than that in the control group during hospitalization (P<0.05). CONCLUSION: Perioperative high-quality nursing for patients undergoing brain tumor surgery can significantly alleviate the adverse psychological states, reduce the complication rate and improve the postoperative self-efficacy and quality of life.

"Turn-Off" Supramolecular Fluorescence Array Sensor for Heavy Metal Ion Identification.

A "turn-off" supramolecular fluorescence array sensor based on the host-guest complexes between fluorescence dyes and cucurbit[n]urils for sensing metal ions was developed. Three fluorescent probes (RhB@Q[7], H33342@2Q[7], and BRE@Q[7]) were used as the sensing units to construct a supramolecular fluorescence array sensor. The binding ability of the metal ions and cucurbituril-dye probes varied; therefore, the probes and metal ions produced different fluorescence responses. When combined with linear discriminant analysis (LDA), the qualitative and quantitative detection of seven metal ions was achieved. In analytical samples, the supramolecular fluorescence array sensor recognized and distinguish seven metal ions. These results provided new research ideas for the rapid analysis and real-time monitoring of different heavy metal ions.

Association Between Eating Speed and Metabolic Syndrome: A Systematic Review and Meta-Analysis.

Objective: This review aimed to systematically summarize and meta-analyze the association between eating speed and metabolic syndrome (MetS). Methods: Following the Preferred Reporting Items for Systematic Reviews, and Meta Analyses (PRISMA) guidelines, four electronic databases (PubMed, Web of Science, MEDLINE, and EMBASE) were searched until March 2021 to identify eligible articles based on a series of inclusion and exclusion criteria. Heterogeneity was examined using I 2 statistics. Using random-effects models, the pooled odds ratios (ORs), and 95% CIs were calculated to evaluate the association between eating speed with MetS and its components, including central obesity, blood pressure (BP), high-density lipoprotein cholesterol (HDL), triglyceride (TG), and fasting plasma glucose (FPG). Results: Of the 8,500 original hits generated by the systematic search, 29 eligible studies with moderate-to-high quality were included, involving 465,155 subjects. The meta-analysis revealed that eating faster was significantly associated with higher risks of MetS (OR = 1.54, 95% CI: 1.27-1.86), central obesity (OR = 1.54, 95% CI: 1.37-1.73), elevated BP (OR = 1.26, 95% CI: 1.13-1.40), low HDL (OR = 1.23, 95% CI: 1.15-1.31), elevated TG (OR = 1.29, 95% CI: 1.18-1.42), and elevated FPG (OR = 1.16, 95% CI: 1.06-1.27) compared to eating slowly. Conclusions: The results of the review indicated that eating speed was significantly associated with MetS and its components. Interventions related to decreasing eating speed may be beneficial for the management of MetS. Systematic Review Registration: https://www.crd.york.ac.uk/prospero/display_record.php?ID=CRD42021242213, identifier: CRD42021242213.

Identification of Risk Factors for Intraoperative Acquired Pressure Injury in Patients Undergoing Neurosurgery: A Retrospective Single-Center Study.

BACKGROUND Risk factors for intraoperative acquired pressure injury (IAPI) in patients undergoing various surgical procedures have been described in previous studies. However, whether risk factors for IAPI in patients undergoing neurosurgery differ remains unknown. The aim of the present study was to explore independent risk factors for IAPI in patients undergoing neurosurgery. MATERIAL AND METHODS Data from 465 patients who underwent neurosurgery between October 2017and December 2018 and who were at high risk of IAPIs were retrospectively analyzed. Independent risk factors for IAPI were evaluated using univariate and multivariate logistic regression models. RESULTS Sixty-nine IAPIs (14.8%) in 465 patients undergoing neurosurgery were assessed. Multivariate logistic regression analyses showed that being overweight (odds ratio [OR] 2.685; 95% confidence interval [CI] 1.206-5.975; P=0.016), prone position (OR 7.502, 95% CI 2.470-22.787. P<0.001), lateral position (OR 15.301, 95% CI 4.903-47.753, P<0.001), use of a head frame (OR 3.716, 95% CI 1.431-9.653, P=0.007), surgical times of 4 to 8 h (OR 7.276, 95% CI 2.249-23.542, P<0.001), and surgical times ≥8 h (OR 173.248, 95% CI 32.629-919.896, P<0.001) all were associated with an increased risk of IAPI in patients undergoing neurosurgery. The factors associated with reduced risk of IAPI were high serum albumin levels (OR 0.099, 95% CI 0.016-0.608, P=0.013) and use of memory sponge pads (OR 0.064, 95% CI 0.020-0.202, P<0.001). CONCLUSIONS The present study indicates that being overweight, prone and lateral positioning, use of a head frame, and longer surgical times are associated with an increased risk of IAPI in patients undergoing neurosurgery. Prospective studies should be conducted to verify these findings and consideration should be given to use of these factors in clinical practice to identify high-risk patients.

Bullying Victimization and Life Satisfaction Among Rural Left-Behind Children in China: A Cross-Sectional Study.

Objectives: This study aimed to evaluate the associations between bullying victimization and life satisfaction in primary school children and also investigate the interactive effects of left-behind status and bullying victimization on life satisfaction. Materials and Methods: Bullying victimization was measured using the Chinese version of the revised Olweus Bully/Victim Questionnaire. Life satisfaction was assessed using the Multidimensional Students' Life Satisfaction Scale (MSLSS). Life satisfaction is composed of five domains, namely, family, school, friends, environment, and self-satisfaction. Left-behind status of rural children was defined as one or both their parents migrating to working in cities. The data were analyzed using Mann-Whitney U tests, Chi-square tests, and multivariate linear and logistic regression analyses. Results: A total of 810 primary school children were involved, of which 8.5% reported bullying victimization, and 44.3% were left-behind children (LBC). We found that bullying victimization was negatively associated with all domains of life satisfaction (all p < 0.05). With further left-behind status-stratified analysis, we found that negative association between bullying victimization and friend satisfaction was more profound in the LBC group than in the non-LBC group [b(SE)= -0.133 (0.03) vs. -0.061 (0.026) for LBC and non-LBC, respectively, p < 0.05]. When further interaction analysis was conducted, we identified interaction effects between left-behind status and bullying victimization on friend satisfaction (p interaction = 0.048). Similar interaction effect between bullying victimization and left-behind status on school satisfaction was also found (p interaction = 0.004). Conclusions: Bullying victimization was associated with low life satisfaction (including lower family, friends, school, self, and environment satisfaction). There were significant interactions between left-behind status and bullying victimization on friend satisfaction, as well as school satisfaction. Left-behind status of children may exaggerate the impact of bullying victimization on friends/school satisfaction rating.

The effect of combined supplementation with vitamin D and omega-3 fatty acids on blood glucose and blood lipid levels in patients with gestational diabetes.

BACKGROUND: To investigate the effects of vitamin D and omega-3 fatty acids on glucose and blood lipid metabolism in gestational diabetes (GDM) women. METHODS: A total of 150 patients with GDM aged 18-40 who were admitted to our hospital from May 2019 to December 2020 were enrolled in this study. The subjects were divided into test and control groups according to whether they took vitamin D and omega-3 fatty acids. The test group took 40,000 IU of vitamin D and 8,000 mg of omega-3 fatty acids twice a day. Comparative analysis of the changes in blood glucose and blood lipid levels of the two groups of patients was performed after 6 weeks. The t-test was used to compare the differences between groups, and the chi-square test was used to assess percentage differences. Repeated measures variance was used to analyze the effects of vitamin D and omega-3 fatty acids on insulin metabolism markers and blood lipid profiles. RESULTS: After adjusting for baseline age and weight, it was found that the fasting blood glucose (FBG), fasting insulin, homeostasis model assessment of insulin resistance (HOMA-IR), triglycerides (TGs), total cholesterol, low-density lipoprotein (LDL), and very-low-density lipoprotein (VLDL) in the test group were decreased by 0.3±0.2 mmol/L, 1.0±0.6 uIU/mL, 0.2±0.1, 0.3±0.1 mmol/L, 0.5±0.2 mmol/L, 1.1±0.4 mmol/L, and 0.03±0.01 mmol/L, respectively, while homeostasis model assessment of beta cell (HOMA-ß) was increased by 0.4±0.1. Compared to the placebo group, the test group's FBG, insulin, HOMA-IR, TGs, total cholesterol, LDL, and VLDL were all significantly decreased, and HOMA-ß was markedly improved. However, no notable statistical difference was observed in the change of high-density lipoprotein (HDL) (P>0.05). CONCLUSIONS: Combined supplementation with vitamin D and omega-3 fatty acids for 6 weeks in patients with GDM can effectively reduce blood sugar and blood lipids, improve HOMA-ß and insulin resistance, and ultimately effectively improve the glucose and lipid metabolism of patients.

MiR-222 promotes the progression of polycystic ovary syndrome by targeting p27 Kip1.

Polycystic ovary syndrome (PCOS) is one of the most complex and common reproductive and endocrinologic disorders in the child-bearing age of women. Recently, miR-222 were reported to be associated with the etiology of PCOS. However, the function of miR-222 during the pathogenesis of PCOS remains unclear. In the present study, we aimed to investigate the role of miR-222 in PCOS. Firstly, miR-222 expression was examined by quantitative real-time PCR (qRT-PCR) in PCOS. The effects of miR-222 on proliferation, apoptosis and cell cycle in KGN cells were analyzed by CCK-8 assay and flow cytometry analysis, respectively. In addition, bioinformatics analysis was used to predict the target genes of miR-222, and dual-luciferase reporter assay was applied to verified the interaction between miR-222 and p27 Kip1 in KGN cells. Moreover, the expressions of p27 Kip1 in KGN cells treated with miR-222 mimics or miR-222 inhibitor were evaluated by qRT-PCR and western blot assays. The results showed that the expression of miR-222 was remarkably upregulated in PCOS tissues compared with corresponding normal tissues. In the gain-of-function and loss-of-function assays, we revealed that miR-222 mimics significantly promoted cell proliferation, while miR-222 inhibitor induced cell apoptosis and cell cycle arrested. Furthermore, p27 Kip1 was identified as a target gene of miR-222, and could be negatively regulated by miR-222 mimics in KGN cells. In conclusion, our findings suggested that miR-222 may promote the progression of PCOS by targeting p27 Kip1.

PhiC31 integrase induces efficient site-specific recombination in the Capra hircus genome.

Streptomyces phage φC31 integrase induces efficient site-specific recombination capable of integrating exogenous genes at pseudo attP sites in human, mouse, rat, rabbit, sheep, Drosophila, and bovine genomes. However, the φC31-mediated recombination between attB and the corresponding pseudo attP sites has not been investigated in Capra hircus. Here, we identified eight pseudo attP sites located in the intron or intergenic regions of the C. hircus genome, and demonstrated different levels of foreign gene expression after φC31 integrase-mediated integration. These pseudo attP sites share similar sequences with each other and with pseudo attP sites in other mammalian genomes, and these are associated with a neighboring consensus motif found in other genomes. The application of the φC31 integrase system in C. hircus provides a new option for genetic engineering of this economically important goat species.

A highly efficient site-specific integration strategy using combination of homologous recombination and the ΦC31 integrase.

The introduction of double-strand breaks (DSBs) at target sites could greatly enhance homologous recombination, and engineered nucleases, such as zinc finger and transcription activator-like effector nucleases, have been successfully developed for making such breaks. In this study, we present a highly efficient site-specific integration strategy based on homologous recombination and ΦC31 integrase. An attB sequence was introduced at the homologous arm of an insertion targeting vector. DSBs at the target locus and donor were then simultaneously generated by the ΦC31 integrase when co-transfected with the donor vector, consequently stimulating homologous recombination. The results demonstrated that our strategy is feasible and the efficiency at the BF4 target site, which we previously identified in the bovine genome, was as high as 93%. The frequency at another site (BF10) was almost two-fold greater in comparison to the vector without homologous arms. This technology requires no sophisticated nuclease design efforts, and the off-target effect is reduced by ΦC31 integrase compared to the use of engineered nucleases, thereby offering a simple and safe way to effectively express a donor gene at a desired locus. This development has great potential value, especially in transgenesis or gene therapy applications.