Advances in understanding the graft healing mechanism: a review of factors and regulatory pathways.

Grafting is a widely used technique for asexual plant reproduction, especially in agriculture and forestry. This procedure is used to shorten the seedling period, improve the structure of scion branches, and help plants adapt to difficult environments. Although grafting has numerous benefits, several obstacles remain to be overcome. The connection between scion and rootstock is regulated by various factors, including phytohormones and molecular mechanisms, which are crucial for graft healing. This review provides an overview of recent advances in the field of grafting, with a specific focus on the factors and regulatory pathways that influence graft healing. The ultimate goal is to aid understanding of how to achieve successful grafting between plants and create desirable grafting chimeras. We provide an overview of the latest developments in plant grafting, covering aspects related to morphology, physiology, and molecular biology. We also discuss research directions in polyploid breeding and long-distance transfer of small molecules in grafted plants.

Canine interleukin-31 binds directly to OSMRß with higher binding affinity than to IL-31RA.

Interleukin-31 (IL-31) is a pro-inflammatory cytokine involved in skin inflammation and tumor progression. The IL-31 signaling cascade is initiated by its binding to two receptors, IL-31 receptor alpha (IL-31RA) and oncostatin M receptor subunit beta (OSMRß). The previous study suggested that human IL-31 (hIL-31) directly interacts with IL-31RA and OSMRß, independently, but the binding ability of hIL-31 to IL-31RA is stronger than to OSMRß. In different to its human ortholog, feline IL-31 (fIL-31) has a higher binding affinity for feline OSMRß. However, the binding pattern of canine IL-31 to its receptors remains to be elucidated. In this study, we purified the recombinant canine IL-31 (rcIL-31) protein and revealed its secondary structure to be mainly composed of alpha-helices. Moreover, in vitro studies show that rcIL-31 has the ability to induce the phosphorylation of signal transducer activator of transcription 3 (STAT3) and STAT5 in DH-82 cells. In the following, the binding efficacies of bioactive rcIL-31 for its individual receptor components have been measured using a flow cytometry assay. The result demonstrates that correctly refolded rcIL-31 binds independently with cIL-31RA and cOSMRß which were expressed on the cell surface. Of note, rcIL-31 has a greater than tenfold higher affinity to OSMRß than to IL-31RA. Additionally, we demonstrated that D1-D4, especially D4 of cOSMRß, is crucial for its binding to cIL-31. Furthermore, this study proved that rcIL-31 has a high binding affinity to the soluble cOSMRß with a KD value of 3.59 × 10-8 M. The results presented in the current study will have a significant implication in the development of drugs or antibodies against diseases induced by cIL-31 signaling.

Risk Factors and Prediction Methods for Endometriosis Combined with Ureteral Stricture Based on Logistic-Regression Analysis.

OBJECTIVE: This study aimed to explore the risk factors of patients with endometriosis (EMS) and ureteral stricture and to establish a prediction model based on logistic-regression analysis. METHODS: The clinical data of 228 EMS patients in Jiaozhou Central Hospital of Qingdao from May 2019 to May 2022 were selected for a retrospective study. According to the results of ureteroscopic biopsy, they were divided into concurrent (n = 32) and nonconcurrent (n = 196) groups. Univariate analysis was performed on the general data and situations of clinical treatment in both groups. Single factor with statistically significant differences was included in unconditional logistic-regression analysis with multiple factors to explore the risk factors of such patients and establish a prediction model. RESULTS: Overt differences were found in previous history of ureter operation (odds ratio (OR) = 3.711, p = 0.006), course of EMS (OR = 3.987, p = 0.007), presence or absence of haematuria (OR = 3.586, p = 0.009) and lateral abdominal pain (OR = 4.451, p = 0.002), and invasion depth of lesion (OR = 7.271, p < 0.001) between the two groups (p < 0.05), without distinct difference in age, menstrual duration, BMI values, history of dysmenorrhea, previous history of drug therapy, smoking history, and drinking history (p > 0.05). Logistic-regression analysis showed that previous history of ureter operation (a1), course of EMS (b2), occurrence of haematuria (c3) and lateral abdominal pain (d4), and invasion depth of lesion ≥5 mm (e5) were risk factors for EMS combined with ureteral stricture (p < 0.05), taking logit (p) = -4.990 + 1.311a1 + 1.383b2 + 1.277c3 + 1.493d4 + 1.984e5 as regression model. The receiver operating characteristic (ROC) curve analysis based on this model showed that the area under the curve (AUC), standard error, and 95% confidence interval (CI) were 0.813, 0.062, and 0.692-0.934, respectively. One hundred EMS patients were re-included, whose values for predictive sensitivity, specificity, and kappa coefficient were 71.40%, 91.10% and 0.615. CONCLUSIONS: Previous history of ureter operation, course of EMS, occurrence of haematuria and lateral abdominal pain, and invasion depth of lesion ≥5 mm were risk factors for EMS combined with ureteral stricture. Therefore, the use of this model has a certain clinical value.

Expression, purification, and biological characterization of recombinant human interleukin-31 protein.

Interleukin-31 (IL-31), belonging to the IL-6 cytokine family, is involved in skin inflammation and pruritus, as well as some tumors' progression. Here, we reported the expression and purification of recombinant human IL-31 (rhIL-31) using a prokaryotic system. This recombinant protein was expressed in the form of inclusion bodies, refolded and purified by size-exclusion chromatography. Circular dichroism analysis revealed that the secondary structure of rhIL-31 was mainly composed of alpha-helix, which is in consistence with the 3D model structure built by AlphaFold server. In vitro studies showed that rhIL-31 exhibited a good binding ability to the recombinant hIL-31 receptor alpha fused with human Fc fragment (rhIL-31RA-hFc) with EC50 value of 16.36 µg/mL in ELISA assay. Meanwhile, flow cytometry demonstrated that rhIL-31 was able to bind to hIL-31RA or hOSMRß expressed on the cell surface, independently. Furthermore, rhIL-31 could induce the phosphorylation of STAT3 in A549 cells. In conclusion, the prepared rhIL-31 in this study possesses the binding ability to its receptors, and can activate the signal pathway of JAK/STAT. Thus, it can be applied in further studies, including investigation of hIL-31-related diseases, structural analysis, and development of therapeutic drugs, and monoclonal antibodies targeting hIL-31.

The Landscape of Accessible Chromatin and Developmental Transcriptome Maps Reveal a Genetic Mechanism of Skeletal Muscle Development in Pigs.

The epigenetic regulation mechanism of porcine skeletal muscle development relies on the openness of chromatin and is also precisely regulated by transcriptional machinery. However, fewer studies have exploited the temporal changes in gene expression and the landscape of accessible chromatin to reveal the underlying molecular mechanisms controlling muscle development. To address this, skeletal muscle biopsy samples were taken from Landrace pigs at days 0 (D0), 60 (D60), 120 (D120), and 180 (D180) after birth and were then analyzed using RNA-seq and ATAC-seq. The RNA-seq analysis identified 8554 effective differential genes, among which ACBD7, TMEM220, and ATP1A2 were identified as key genes related to the development of porcine skeletal muscle. Some potential cis-regulatory elements identified by ATAC-seq analysis contain binding sites for many transcription factors, including SP1 and EGR1, which are also the predicted transcription factors regulating the expression of ACBD7 genes. Moreover, the omics analyses revealed regulatory regions that become ectopically active after birth during porcine skeletal muscle development after birth and identified 151,245, 53,435, 30,494, and 40,911 peaks. The enriched functional elements are related to the cell cycle, muscle development, and lipid metabolism. In summary, comprehensive high-resolution gene expression maps were developed for the transcriptome and accessible chromatin during postnatal skeletal muscle development in pigs.

Integrated Proteotranscriptomics Reveals Differences in Molecular Immunity between Min and Large White Pig Breeds.

Long-term selection or evolution is an important factor governing the development of disease resistance in pigs. To better clarify the molecular mechanisms underlying different levels of disease resistance, we used transcriptomics and proteomics analysis to characterize differences in the immunities between six resistant (Min pig) and six susceptible (Large White, LW) pigs which were raised in the same environment. A total of 135 proteins and 791 genes were identified as being differentially expressed between the Large White and Min pig groups. Protein expression clustering and functional analysis revealed that proteins related to immune system process, humoral immune response, the B cell receptor signaling pathway, lymphocyte-mediated immunity, and innate immune responses were more highly expressed in Min pigs. Transcriptome gene set enrichment analysis was used to reveal that pathways of cell adhesion molecules and antigen processing and presentation are significantly enriched in Min pigs. Integrated proteomics and transcriptomics data analysis identified 16 genes that are differentially expressed at both the mRNA and protein levels. In addition, 13 out of these 16 genes were related to the quantitative trait loci of immune diseases, including neural EGFL-like 2 (NELL2) and lactate dehydrogenase B (LDHB), which are involved in innate immunity. Correlation analysis between the genes/proteins and cytokines shows upregulated proteins in LW pigs in association with immunosuppressive/pro-inflammatory cytokines, such as interleukin (IL) 10, IL6, and tumor necrosis factor alpha. This was further validated using parallel reaction monitoring analysis. In summary, we discovered several potential candidate pathways and key genes/proteins involved in determining differences in disease resistance between the two studied pig breeds, which could provide new insights into the breeding of pigs for disease resistance.

Study of ultrasound-guided ropivacaine combined with butorphanol continuous paravertebral block to prevent pain syndrome by evaluating ccl2 gene expression after radical mastectomy.

This study aimed to investigate the clinical effect of ultrasound-guided ropivacaine combined with butorphanol continuous paravertebral block in preventing postoperative pain syndrome of breast cancer. For this purpose, 100 women treated for breast cancer from April 2018 to July 2019 were enrolled as research objects. Surgical procedures included local sentinel lymph node biopsy, mastectomy, sentinel lymph node biopsy for mastectomy, modified radical mastectomy, and implantation. The selected patients were randomly divided into two groups: control group (routine operation anesthesia; n = 50) and observation group (ultrasound-guided thoracic paravertebral block before induction of ropivacaine+butorphanol anesthesia; n = 50). The Real-time PCR technique was performed to evaluate CCL2 gene expression. VAS scores were recorded during the postoperative period. Compared with the control group, the observation group had lower VAS scores at six h, 24h, and 48h (P<0.05). The pain effect of the observation group was less than that of the control group. The observation group had better analgesic effects after anesthesia. The observation group had a lower incidence of pain syndrome at the 6th, 8th, and 12th months (P<0.05), and the incidence of pain syndrome in the two groups decreased with the extension of time. The observation group had lower levels of related factors (P<0.05), and the observation group had lower traumatic stress responses. The protein expression of IL-6, IL-17, and CRP in the observation group was lower than that in the control group (P<0.05). The results of CCL2 gene expression also showed that gene expression in the control group increased significantly (P=0.0047). Since the expression of this gene is one of the factors that stimulate pain signals in the body, the method used in the present study was able to reduce the amount of pain significantly. Therefore, the combination of ropivacaine combined with butorphanol ultrasound-assisted paravertebral block can reduce the intensity of postoperative pain in patients with breast cancer surgery, decrease the incidence of pain syndrome, and increase pain tolerance.

Development and characterization of a novel mouse anti-canine oncostatin M receptor beta monoclonal antibody.

Oncostatin M receptor beta (OSMRß) mediates signaling of Oncostatin M (OSM) and interleukine-31 (IL-31), two key cytokines involved in many important biological processes including inflammation and cancer progression. More importantly, OSMRß might be a potential biomarker and therapeutic target for some diseases, such as inflammatory bowel disease, pruritus and ovarian cancer. In this study, soluble recombinant canine OSMRß (cOSMRß) was experimentally expressed as a native antigen to develop an effective cOSMRß-specific monoclonal antibody (mAb), 2O2, using hybridoma technology. It was demonstrated that 2O2 is able to detect OSMRß expressed on cell surface using immunofluorescence assay (IFA) and flow cytometry (FACS). This mAb exhibits very high binding affinity to cOSMRß with the KD and half-maximal effective concentration (EC50) values of 2.49 nM and 96.96 ng/ml, respectively. Meanwhile, it didn't show any cross-relativities with feline OSMRß (fOSMRß) and human OSMRß (hOSMRß). Moreover, we determined the binding epitope of 2O2, which localizes in the domain VI (DVI, amino acids 623-734) of cOSMRß. In conclusion, this novel mAb, 2O2, can be used in immunoassays, including IFA, FACS and enzyme-linked immunosorbent assay (ELISA) to facilitate studies in dogs.

Comparison of the effects of different mechanical ventilation modes on the incidence of ventilation-associated pneumonia: a case study of patients undergoing thoracic surgery.

OBJECTIVE: To investigate the effects of different mechanical ventilation modes on the incidence of ventilator-associated pneumonia (VAP) in patients undergoing thoracic surgery. METHODS: From June 2019 to December 2021, the clinical data of 96 patients undergoing thoracic surgery in Cangzhou Central Hospital were retrospectively analyzed. A total of 44 patients who underwent constant flow mode were included in the control group (CG), and 52 patients who underwent auto flow mode were included in the observation group (OG). The respiratory mechanics, hemodynamics, blood gas analysis and serum levels of lung injury markers at different time points were compared between the two groups, and the incidence of VAP was analyzed. RESULTS: At 1 hour and 4 hours of ventilation, the peak airway pressure (Ppeak), Pmean mean airway pressure (Pmean) and airway resistance (Raw) of the OG were lower than those of the CG, and the dynamic lung compliance (Cdyn) was higher than that of the CG (P<0.05). There were no statistically significant differences in mean arterial pressure (MAP), heart rate (HR), blood oxygen saturation (SpO2), PH, arterial partial pressure of oxygen (PaO2), arterial partial pressure of carbon dioxide (PaCO2) between the OG and CG at 1 hour and 4 hours of ventilation respectively (P>0.05). The serum levels of pulmonary surfactant associated protein A (SP-A), human Clara cell protein (CC16) and serum ferritin (SF) in the OG were lower than those in the CG (P<0.05). The incidence of VAP in the OG (3.85%) was lower than that in the CG (15.91%) (P<0.05). CONCLUSION: In mechanical ventilation, auto flow mode can reduce the incidence of VAP, improve respiratory mechanics, and reduce lung injury in patients undergoing thoracic surgery, but has no significant effect on hemodynamics and blood gas analysis.

Experimental studies of hydrothermal liquefaction of kitchen waste with H+, OH- and Fe3+ additives for bio-oil upgrading.

Kitchen waste (KW) has gradually become a prominent problem in municipal solid waste treatment. Hydrothermal liquefaction (HTL) is a promising method used to make fuel oil from food and KW. However, the upgrading of bio-oil is particularly important for the sake of industrial reuse. In this study, the KW from university restaurants was subjected to HTL experiments in order to study theoretical feasibility. With the change of conversion temperature and residence time, the optimal conversion working conditions in this study were determined according to the quality and yield of the bio-oil. Moreover, the bio-oil upgrading effects of different additives (hydrogen chloride, sodium hydroxide, and iron(III) chloride) on the HTL of KW were studied. Alkaline additives have an inhibitory effect on the bio-oil yield and positive effect on coke yield. Acidic additives and iron (Fe)-containing additives can promote bio-oil yield. As an important aspect of upgrading, the effect on the nitrogen content of bio-oil with additives was revealed. The alkaline and Fe-containing additives have little effect on reducing the viscosity of the bio-oil while with the appropriate ratio (2.5 mol•kg-1) of acidic additives to the raw material, the static and dynamic fluidity of the oil phase products are reduced to about 0.1 Pa•s.

MiR-21-3p Centric Regulatory Network in Dairy Cow Mammary Epithelial Cell Proliferation.

MicroRNA-mediated gene regulation is important for the development of the mammary gland and the lactating process. A previous study has shown that the expression of microRNA-21 (miR-21) is different in the dry and early lactation period of the dairy cow mammary gland, but the molecular mechanisms underlying the lactation cycle are not fully understood. Here, the function of miR-21-3p on bovine mammary gland epithelial cells (BMECs) was detected by MTT assay and flow cytometry analysis, which showed that miR-21-3p significantly promoted the cell viability and proliferation. Then, the regulating mechanism of miR-21-3p on cell viability and proliferation was elucidated. Dual luciferase assay, RT-qPCR, and Western blot results revealed that IGFBP5 was a target gene of miR-21-3p. It was known that lncRNA could act as a competing endogenous RNA to sequester miRNAs and reduce the regulatory effect of miRNA-targeted genes. Based on our previous lncRNA-seq data and bioinformatics analysis, lncRNA NONBTAT017009.2 was potentially associated with miR-21-3p, and its expression was specifically inhibited with the transfection of miR-21-3p mimic into BMECs. Inversely, the overexpression of NONBTAT017009.2 significantly decreased the expression level of miR-21-3p in BMECs, while the expression of IGFBP5, the target gene of miR-21-3p, was significantly upregulated. In addition, the promoter region of miR-21 contained two STAT3 binding sites, and the dual luciferase reporter assays revealed that the overexpression of STAT3 significantly reduced the promoter activity of miR-21, implying that the transcription factor STAT3 may act as an upstream regulator affecting the regulation process of miR-21-3p. The overexpression of STAT3 significantly inhibited the expression of miR-21-3p, while the mRNA expression of IGFBP5 was significantly increased compared with the control group. Besides, there are no STAT3 binding sites in the promoter region of IGFBP5 as we predicted by gene-regulation and JASPAR software. Therefore, it could infer that STAT3 might regulate the expression of IGFBP5 by miR-21-3p. Taken together, these results established a regulatory network of miR-21-3p to illustrate the regulating mechanism on promoting cow mammary epithelial cell proliferation.

Hypomethylation of the GSTM1 promoter is associated with ovarian endometriosis.

STUDY QUESTION: Is the methylation status of the glutathione S-transferase M1 (GSTM1) promoter region altered in patients with ovarian endometriosis, and does this affect the expression of GSTM1 in their endometrial tissues? SUMMARY ANSWER: The promoter region of GSTM1 was significantly hypomethylated in the ectopic and eutopic endometrium of patients with ovarian endometriosis and this was associated with higher expression of GSTM1 mRNA. WHAT IS KNOWN ALREADY: GSTM1, a member of the glutathione S-transferase family, is primarily known as a detoxification enzyme, but it has also been shown to negatively regulate apoptosis-related signalling cascades through protein-protein interactions with apoptosis signal-regulating kinase-1. STUDY DESIGN, SIZE, DURATION: This is a case-control study between September 2013 and December 2016, involving 65 patients with ovarian endometriosis and 53 women without endometriosis. We analysed the methylation status and expression levels of GSTM1 in the ectopic and eutopic endometrium of patients with ovarian endometriosis and the endometrium of women without endometriosis. In addition, we collected endometrial samples from 12 women without endometriosis for endometrial epithelial cell cultures. PARTICIPANTS/MATERIALS, SETTING, METHODS: Methylation levels of the GSTM1 promoter region in the ectopic and eutopic endometrial tissues of patients with ovarian endometriosis and the endometrial tissues of women without endometriosis were analysed by pyrosequencing. The expression of GSTM1 mRNA and protein in endometrial tissues was investigated by RT-qPCR and immunohistochemistry, respectively. Primary cell culture, gene transfection, Cell Counting Kit-8 assay and flow cytometry were used to analyse the effect of GSTM1 on viability and apoptosis in endometrial epithelial cells. MAIN RESULTS AND THE ROLE OF CHANCE: Compared with that in the endometrium of women without endometriosis, the GSTM1 promoter region was significantly hypomethylated in the ectopic and eutopic endometrium of patients with ovarian endometriosis. Additionally, GSTM1 mRNA and protein levels were significantly higher in the ectopic and eutopic endometrium than in the control endometrium. Moreover, the methylation levels of the GSTM1 promoter region were significantly negatively correlated with the mRNA expression of GSTM1. Furthermore, in vitro results suggested that the over-expression of GSTM1 could significantly increase viability and inhibit apoptosis in endometrial epithelial cells following hormone treatment and withdrawal. LIMITATIONS, REASONS FOR CAUTION: Due to restrictions in the isolation and culture of pure populations of endometrial epithelial cells, as well as limitations in the number of passages possible in primary cells, we could not explore the underlying molecular mechanism by which GSTM1 modulates apoptosis in endometrial cells. WIDER IMPLICATIONS OF THE FINDINGS: This study provides new evidence to support the notion that endometriosis may be an epigenetic disease. STUDY FUNDING/COMPETING INTEREST(S): This work was supported by grants from the Natural Science Foundation of Hebei Province (Grant number: H2018206200) and the Department of Education of Hebei Province (Grant number: CXZZBS2017114). The authors have no conflicts of interest to declare.

Genome-wide identification of RNA editing in seven porcine tissues by matched DNA and RNA high-throughput sequencing.

BACKGROUND: RNA editing is a co/posttranscriptional modification mechanism that increases the diversity of transcripts, with potential functional consequences. The advent of next-generation sequencing technologies has enabled the identification of RNA edits at unprecedented throughput and resolution. However, our knowledge of RNA editing in swine is still limited. RESULTS: Here, we utilized RES-Scanner to identify RNA editing sites in the brain, subcutaneous fat, heart, liver, muscle, lung and ovary in three 180-day-old Large White gilts based on matched strand-specific RNA sequencing and whole-genome resequencing datasets. In total, we identified 74863 editing sites, and 92.1% of these sites caused adenosine-to-guanosine (A-to-G) conversion. Most A-to-G sites were located in noncoding regions and generally had low editing levels. In total, 151 A-to-G sites were detected in coding regions (CDS), including 94 sites that could lead to nonsynonymous amino acid changes. We provide further evidence supporting a previous observation that pig transcriptomes are highly editable at PRE-1 elements. The number of A-to-G editing sites ranged from 4155 (muscle) to 25001 (brain) across the seven tissues. The expression levels of the ADAR enzymes could explain some but not all of this variation across tissues. The functional analysis of the genes with tissue-specific editing sites in each tissue revealed that RNA editing might play important roles in tissue function. Specifically, more pathways showed significant enrichment in the fat and liver than in other tissues, while no pathway was enriched in the muscle. CONCLUSIONS: This study identified a total of 74863 nonredundant RNA editing sites in seven tissues and revealed the potential importance of RNA editing in tissue function. Our findings largely extend the porcine editome and enhance our understanding of RNA editing in swine.

Aberrant methylation of the IL-12B promotor region contributes to the risk of developing ovarian endometriosis.

Studies have shown that aberrant expression of IL-12p40, which is encoded by the interleukin-12B (IL-12B) gene, may be involved in the development of endometriosis. In this study, we investigated the role of aberrant methylation of the IL-12B promoter region and its associated expression in the development of ovarian endometriosis. By using pyrosequencing, we analyzed the methylation level of the IL-12B promoter region in eutopic and ectopic endometrium of patients with ovarian endometriosis and normal endometrium of control women. The expression of IL-12B mRNA was detected by quantitative real-time PCR. The results showed that the methylation level of the IL-12B promoter region in ectopic and eutopic endometrium of patients with ovarian endometriosis was significantly lower than that in endometrium of women without endometriosis ( p < 0.001 and p = 0.041, respectively). In contrast, mRNA levels were significantly increased in ectopic and eutopic endometrium of patients with ovarian endometriosis compared to those in endometrium of women without endometriosis ( p < 0.001 and p = 0.042, respectively). Correlation analysis showed that the methylation level of the IL-12B promoter region was negatively correlated with mRNA levels of IL-12B ( p < 0.001). Our data suggested that aberrant methylation of the IL-12B promoter region may be responsible for aberrant IL-12B mRNA expression in endometrium tissue of women, which may be associated with the development of ovarian endometriosis in northern Chinese women.

BMP4 Cooperates with Retinoic Acid to Induce the Expression of Differentiation Markers in Cultured Mouse Spermatogonia.

Spermatogenesis is sustained by the proliferation and differentiation of spermatogonial stem cells (SSCs). However, the molecules controlling these processes remain largely unknown. Here, we developed a simplified high concentration serum-containing system for the culture of mouse SSCs. Analysis of SSCs markers and transplantation results revealed that the cultured spermatogonia retained stem cell characteristics after long-term in vitro propagation. Using this culture system, the expression and function of bone morphogenetic protein 4 (BMP4) were explored. Immunostaining showed that BMP4 was predominantly expressed in germ cells and that its level increased as spermatogenesis progresses. BMP4 receptors BMPR1A and BMPRII were present in spermatogonia, spermatocytes, and round spermatids. Moreover, despite the mRNAs of these two genes being present in mouse Sertoli cells, only BMPRII was detected by using Western blotting assays. While exogenous BMP4 by itself did not induce the expression of Stra8 and c-Kit, two marker genes of differentiating spermatogonia, a significant cooperative effect of BMP4 and retinoic acid (RA) was observed. Moreover, pretreatment of cultured spermatogonia with the BMP4 antagonist Noggin could inhibit RA-induced expression of these two marker genes. In conclusion, BMP4 may exert autocrine effects and act cooperatively with RA to induce the differentiation of spermatogonia in vivo.

Mensenchymal stem cells can delay radiation-induced crypt death: impact on intestinal CD44(+) fragments.

Intestinal stem cells are primitive cells found within the intestinal epithelium that play a central role in maintaining epithelial homeostasis through self-renewal and commitment into functional epithelial cells. Several markers are available to identify intestinal stem cells, such as Lgr5, CD24 and EphB2, which can be used to sort intestinal stem cells from mammalian gut. Here, we identify and isolate intestinal stem cells from C57BL/6 mice by using a cell surface antigen, CD44. In vitro, some CD44(+) crypt cells are capable of forming "villus-crypt"-like structures (organoids). A subset strongly positive for CD44 expresses high levels of intestinal stem-cell-related genes, including Lgr5, Bmi1, Hopx, Lrig1, Ascl2, Smoc2 and Rnf43. Cells from this subset are more capable of developing into organoids in vitro, compared with the subset weakly positive for CD44. However, the organoids are sensitive to ionizing irradiation. We investigate the specific roles of mesenchymal stem cells in protecting organoids against radiation-induced crypt death. When co-cultured with mesenchymal stem cells, the crypt domains of irradiated organoids possess more proliferative cells and fewer apoptotic cells than those not co-cultured with mesenchymal stem cells. Cd44v6 continues to be expressed in the crypt domains of irradiated organoids co-cultured with mesenchymal stem cells. Our results indicate specific roles of mesenchymal stem cells in delaying radiation-induced crypt death in vitro.

Copy number variation-based genome wide association study reveals additional variants contributing to meat quality in Swine.

Pork quality is important both to the meat processing industry and consumers' purchasing attitude. Copy number variation (CNV) is a burgeoning kind of variants that may influence meat quality. In this study, a genome-wide association study (GWAS) was performed between CNVs and meat quality traits in swine. After false discovery rate (FDR) correction, a total of 8 CNVs on 6 chromosomes were identified to be significantly associated with at least one meat quality trait. All of the 8 CNVs were verified by next generation sequencing and six of them were verified by qPCR. Only the haplotype block containing CNV12 is adjacent to significant SNPs associated with meat quality, suggesting the effects of those CNVs were not likely captured by tag SNPs. The DNA dosage and EST expression of CNV12, which overlap with an obesity related gene Netrin-1 (Ntn1), were consistent with Ntn1 RNA expression, suggesting the CNV12 might be involved in the expression regulation of Ntn1 and finally influence meat quality. We concluded that CNVs may contribute to the genetic variations of meat quality beyond SNPs, and several candidate CNVs were worth further exploration.

Protein disulfide isomerase homolog TrPDI2 contributing to cellobiohydrolase production in Trichoderma reesei.

The majority of the cysteine residues in the secreted proteins form disulfide bonds via protein disulfide isomerase (PDI)-mediated catalysis, stabilizing the enzyme activity. The role of PDI in cellulase production is speculative, as well as the possibility of PDI as a target for improving enzyme production efficiency of Trichoderma reesei, a widely used producer of enzyme for the production of lignocellulose-based biofuels and biochemicals. Here, we report that a PDI homolog, TrPDI2 in T. reesei exhibited a 36.94% and an 11.81% similarity to Aspergillus niger TIGA and T. reesei PDI1, respectively. The capability of TrPDI2 to recover the activity of reduced and denatured RNase by promoting refolding verified its protein disulfide isomerase activity. The overexpression of Trpdi2 increased the secretion and the activity of CBH1 at the early stage of cellulase induction. In addition, both the expression level and redox state of TrPDI2 responded to cellulase induction in T. reesei, providing sustainable oxidative power to ensure cellobiohydrolase maturation and production. The results suggest that TrPDI2 may contribute to cellobiohydrolase secretion by enhancing the capability of disulfide bond formation, which is essential for protein folding and maturation.

Cucurbitacin E suppresses cytokine expression in human Jurkat T cells through down-regulating the NF-κB signaling.

Cucurbitacin E (CucE), a triterpenoid isolated from Cucurbitaceae plants, has been shown to possess an anti-inflammatory or immunosuppressive activity in vitro and in vivo, yet the underlying mechanism has been incompletely understood. The aim of the present study was to explore its effect on cytokine expression and the underlying mechanism in human Jurkat T cells as a cellular model. The results showed that CucE significantly inhibited the production of interleukin-2, tumor necrosis factor-α, and interferon-γ in culture medium of cells treated with phorbol 12,13-dibutyrate (PDB) plus ionomycin (Ion). Furthermore, the mRNA levels of these cytokines in activated Jurkat T cells were also decreased upon CucE treatment, suggesting a potential modulatory effect on the critical signaling pathways for cytokine expression, including nuclear factor-κB (NF-κB) or mitogen-activated protein kinases (MAPKs). In support of its effect on the NF-κB signaling pathway, CucE decreased the phosphorylation levels of inhibitor of κB (IκB) and NF-κB/p65 in PDB + Ion-stimulated cells. Further supporting this, the nuclear translocation of NF-κB/p65 was significantly suppressed in response to PDB plus Ion stimulation in the presence of CucE. The phosphorylation of p38MAPK, c-Jun N-terminal kinase (JNK), and Erk1/2, however, was not decreased or slightly increased at some time points by CucE treatment. Collectively, these data suggest that CucE may exhibit immunosuppressive effect by attenuating critical cytokine expression through down-regulating the NF-κB signaling pathway.

Transversus abdominis plane block versus local anaesthetic wound infiltration for postoperative analgesia: A systematic review and meta-analysis.

BACKGROUND: Transversus abdominis plane (TAP) block and local anaesthetic wound infiltration can provide effective pain relief at the wound site after surgery. However, the relative efficacy of two techniques for postoperative analgesia remains controversial. METHODS: We searched PUBMED, EMBASE and CENTRAL databases for randomized controlled trials (RCTs) comparing TAP block with wound infiltration for pain relief after surgery. The primary outcomes were pain scores at rest and on movement at 1, 8 and 24 hours postoperatively and cumulative morphine consumption over 24 hours. The secondary outcomes were time to first rescue analgesic, number of rescue analgesic use and opioids-related side-effects. RESULTS: Nine RCTs with a total of 500 participants were included. TAP block was associated with significant lower rest and dynamic pain scores at 8 hour [MD = -1.08, 95% CI (-1.89-0.26), P = 0.009] and 24 hour [MD = -0.83, 95% CI (-1.60, -0.06), P = 0.03] postoperatively than wound infiltration, but no significant difference was found at 1 hour [MD = -0.94, 95% CI (-1.97, 0.09), P = 0.08] postoperatively. In adults, TAP block significantly reduced 24-hour overall morphine consumption by 3.85 mg [MD = -3.85, 95% CI (-7.47, -0.22), P = 0.04] compared with wound infiltration. Subgroup analysis showed that adults received TAP block appeared to have lower rest pain scores at 24 hour than children (P = 0.008). CONCLUSION: TAP block provides superior analgesia compared with wound infiltration in the setting of a multimodal analgesic regimen. Subgroup analysis indicated that adults may have benefits additional to the analgesic effect than children.