Family intimacy and adaptability and non-suicidal self-injury: a mediation analysis.

BACKGROUND: Current research has been focusing on non-suicidal self-injury (NSSI) behaviors among adolescents with depression. Although family intimacy and adaptability are considered protective factors for NSSI, evidence supporting this relationship is lacking. OBJECTIVE: This study aims to examine the mechanisms operating in the relationship between family intimacy and adaptability and NSSI behaviors among adolescents. METHODS: A self-administered general demographic information questionnaire, the Behavioral Functional Assessment Scale for Non-Suicidal Self-Injury, the Family Intimacy and Adaptability Scale, the Connor-Davidson Resilience Scale, and the Self-Assessment of Depression Scale were distributed among adolescents with depression in three tertiary hospitals in Jiangsu Province. RESULTS: The relationship between family intimacy and adaptability and NSSI was assessed among 596 adolescents with depression. The results revealed the following: (1) Family intimacy and adaptability were negatively correlated with NSSI behavior. (2) Psychological resilience and depression levels acted as chain mediators in the relationship between family intimacy and adaptability and NSSI behavior. CONCLUSIONS: Enhancing psychological resilience, controlling depressive symptoms, and reducing depression severity among adolescents by improving their family intimacy and adaptability are conducive to preventing and mitigating their NSSI behaviors.

Perinatal maternal factors influencing postpartum feeding practices at six weeks.

OBJECTIVE: Despite the World Health Organization's recommendation of exclusive breastfeeding for the initial 6 months, breastfeeding rates decline within the first 6 weeks after delivery. This study aimed to (1) investigate the breastfeeding rate at 6 weeks postpartum and (2) explore the influence of perinatal factors on feeding patterns at 6 weeks postpartum. METHOD: A total of 635 participants were enrolled from February to August 2023 at outpatient clinics in three tertiary hospitals in Nantong City. Variables were collected through questionnaires during the third trimester of pregnancy, including demographic information, pregnancy stress, anxiety, depression, sleep, and resilience. At 6 weeks postpartum, information regarding feeding patterns, delivery and postpartum situations, postpartum stress, anxiety, depression, sleep, and resilience was gathered. Initial single-factor analyses were conducted using feeding pattern as the dependent variable, and variables with significance were chosen as independent variables. The disordered multi-classification logistic regression model was then established using the stepwise forward method. RESULTS: Within the first 6 weeks, 35.28% (224/635) of postpartum women exclusively breastfed their infants. Factors influencing exclusive breastfeeding and formula feeding at 6 weeks postpartum included breast pain, sleep quality, mental resilience, difference between postpartum and late pregnancy anxiety, insufficient milk supply, and maternal herself caring for the infant (P < 0.05). Factors influencing the transition from exclusive to partial breastfeeding were insufficient milk supply and maternal herself caring for the infant (P < 0.05). CONCLUSION: The study reveals a relative low rate of exclusive breastfeeding in China's first 6 weeks postpartum, along with a comparison of perinatal factors affecting three different feeding patterns. Our findings may contribute additional evidence to the association between perinatal factors and feeding patterns. This study guides healthcare professionals in developing strategies to promote exclusive breastfeeding and improve personalized counseling for exclusive breastfeeding and mental health.

Fluorine accumulation characteristics of 85 tea tree (Camellia sinensis) varieties and its potential risk assessment.

Tea tree is a fluorine (F)-enriched plant, leading to much concern about the safety of drinking tea from tea tree (Camellia sinensis (L.) Kuntze). Tea tree is a perennial leaf-harvested crop, and tea production in China is generally categorized as spring tea, summer tea and autumn tea in its annual growth rounds. However, the seasonally dynamic changes of F content and accumulation in the leaves and its drinking safety are poorly understood. In this study, 85 tea varieties cultivated under the same conditions were investigated to analyze the seasonal variation of F content and it's relationships with F accumulation, aluminum (Al), calcium (Ca) and manganese (Mn) and hazard quotient (HQ) in young leaves (one bud and two leaves, YL) and mature leaves (canopy leaves, ML). The average F contents and accumulations were 350 mg kg-1 and 203 g ha-1 in YL, and they were 2451 mg kg-1 and 2578 g ha-1 in ML, respectively, with F mainly accumulated in ML. As the growing season progresses, the F content showed a gradual increase in YL, while a decrease in ML, inferring that F may be redistributed from mature leaves to young leaves. Additionally, the F content was quite different among tea varieties which are suitable for processing oolong tea, green tea, and black tea, with higher F accumulation in oolong tea varieties than in green and black tea varieties. Moreover, F content and accumulation could be obviously affected by the geographical origin of the tea tree varieties, with significantly higher F content in the varieties from F rich fluorite belts than other regions. Furthermore, F content and accumulation showed a significant positive correlation with the content of Al and Mn (p < 0.05). Based on a daily tea consumption of 8.7 g, the HQ was investigated to show that the proportion of tea leaves with HQ<1 made from spring, summer and autumn tender leaves of 85 varieties was 100 %, 90.6 % and 50.6 %, respectively, indicating that the tea with the best drinking safety comes from spring, followed by summer, and then autumn. This result suggests that it could be necessary to avoid planting tea trees in fluorite mining areas, choose low F tea tree varieties, and control the tenderness of fresh leaves in order to ensure the safety of tea drinking.

The mediating role of social support and resilience between self-efficacy and prenatal stress: a mediational analysis.

BACKGROUND: Prenatal stress is a highly prevalent mental disorder experienced by pregnant women. This study assessed the prevalence and influencing factors of prenatal stress and investigated the mediating role of social support and resilience between self-efficacy and prenatal stress among pregnant women in China. METHODS: A convenience sample comprising 1071 pregnant women from three hospitals in Nantong, Jiangsu Province, China, was recruited between February and June 2023. These participants completed a set of general survey questionnaires and were assessed using the Pregnancy Pressure Scale, Perceived Social Support Scale, the 10-item Connor-Davidson Resilience Scale, and the Chinese version of the General Self-Efficacy Scale. Furthermore, a hierarchical multiple regression model was employed to investigate the relevant factors and mediators of prenatal stress symptoms. A structural equation model was used to examine the mediating role of social support and resilience in the relationship between self-efficacy and prenatal stress. RESULTS: The results of the multivariate regression analysis indicated significant associations between prenatal stress and parity, self-efficacy, social support, and resilience (P < 0.001). Self-efficacy accounted for 35.33% of the total effect, with a direct effect of -2.5306 (95% confidence interval [CI]: -4.0309 to -1,0303). Further examination through mediation analysis revealed the mediating roles of social support and resilience in the relationship between self-efficacy and prenatal stress. The mediating effect of social support was - 1.5933 (95% CI: -2.2907 to -0.9496), accounting for 22.24% of the total effect. Similarly, resilience exhibited a mediating effect of -3.0388 (95% CI: -4.3844 to -1.7135), accounting for 42.43% of the total effect. CONCLUSION: The mediation analysis revealed that among pregnant women in China, the influence of self-efficacy on prenatal stress is channelled through social support and resilience. Therefore, enhancing social support, resilience, and self-efficacy might alleviate prenatal stress.

Translation and validation of the Chinese version of the maternal postpartum stress scale.

OBJECTIVE: To translate the Maternal Postpartum Stress Scale (MPSS) into Chinese and validate its psychometric properties in postpartum women. METHODS: A total of 406 postpartum women were recruited from six hospitals in Nantong, Jiangsu Province, China. Cronbach's α co-efficient, split-half reliability, and test-retest reliability were used to evaluate the reliability of the translated scale. Exploratory factor analysis (EFA) and confirmatory factor analysis (CFA) were used to evaluate the structural validity of the scale. The Edinburgh Postnatal Depression Scale, Depression Anxiety Stress Scale-21 anxiety dimension, and Perceived Stress Scale were used as calibration scales to measure the correlation of MPSS. All data were analyzed using SPSS 25.0 and Amos 24.0. RESULTS: The Cronbach's α co-efficient of the Chinese version of MPSS and its three dimensions were 0.940 and 0.882-0.911, respectively. The split-half reliability was 0.825, and the test-retest reliability was 0.912. The scale's content validity index was 0.926. Three common factors were extracted from the EFA. The CFA validated the explored 3-factor structure, and the indicators were fitted well (χ2/Df = 2.167, comparative fit index = 0.918, Tucker-Lewis index = 0.907, incremental fit index = 0.919, and root mean square error of approximation = 0.075). CONCLUSION: The translated Chinese version of MPSS had suitable reliability and validity in assessing postpartum stress in Chinese women. The translated scale can also help with the early identification of postpartum stress and provide a scientific basis for the formulation of early personalized intervention measures. Overall, the scale has certain clinical value and practical significance for enhancing the physical and mental health of postpartum women. However, future studies including large, diverse populations are warranted.

The effect of perceived social support on postpartum stress: the mediating roles of marital satisfaction and maternal postnatal attachment.

BACKGROUND: Multiple factors may be responsible for the development of postpartum stress, including perceived social support, marital satisfaction, and maternal postnatal attachment. However, the underlying mediation mechanisms remain unclear. This study examined the complex relationships between perceived social support and postpartum stress among Chinese women. METHODS: A convenience sample comprising 406 postpartum women was recruited from six hospitals in Nantong, Jiangsu Province, China. The participants completed general survey questionnaires and were evaluated using the Maternal Postpartum Stress Scale, the Perceived Social Support Scale, the Maternal Postnatal Attachment Scale, and the Marital Satisfaction Scale. Furthermore, we evaluated the relationship between postpartum stress and the various influencing factors by performing a multiple linear regression analysis. The potential mediating roles of marital satisfaction and maternal and infant attachment in the association between perceived social support and postpartum stress were explored by performing a mediation analysis. RESULTS: According to the multivariate regression analysis, perceived social support, marital satisfaction, and maternal postnatal attachment contributed to postpartum stress levels (P < 0.05). The mediation analysis revealed that marital satisfaction and maternal postnatal attachment played parallel mediating roles in the association between perceived social support and postpartum stress, and the mediating effect of marital satisfaction was - 0.1125 (95% confidence interval [CI]: -0.1784 to -0.0520), accounting for 33.20% of the total effect, and the mediating effect of maternal postnatal attachment was - 0.0847 (95% CI: -0.1304 to -0.0438), accounting for 25.00% of the total effect. CONCLUSION: Our study revealed that perceived social support could influence postpartum stress not only through direct effect (41.80% of the total effect), but also through the indirect effect (mediation effect) of marital satisfaction and maternal postnatal attachment (58.20% of the total effect), suggesting that improving postpartum women's social support, enhancing maternal and infant attachment, and improving their marital satisfaction could help lower postpartum stress.

Effects of gene knockdown of CNP on ventricular remodeling after myocardial ischemia-reperfusion injury through NPRB/Cgmp signaling pathway in rats.

This study aimed to explore effects of CNP on ventricular remodeling following myocardial ischemia-reperfusion (I/R) injury through the NPRB/cGMP signaling pathway. Rat cardiomyocytes were assigned into: control, I/R, I/R + CNP, and I/R + 8-Br-cGMP groups. ELISA, qRT-PCR, and Western blotting were used to detect cGMP content and expression, respectively. After model establishment of I/R rats, normal control, CNP-/- control, I/R, and CNP-/- groups were set. Indexes of heart were detected using echocardiography and hemodynamics. ELISA was used to measure serum CNP, cGMP, LDH, cTn I, CK-MB, TNF-α, and IL-6 levels. Myocardial infarct was identified by TTC staining, and apoptosis conditions by TUNEL staining. QRT-PCR and Western blotting were adopted to detect expressions of CNP, NPRB, cGMP, and apoptosis-related genes. Compared with control group, cGMP contents and expression in the I/R, I/R + CNP and I/R + 8-Br-cGMP groups were decreased. Levels of LVEDV, LVESV, LVDS, LVDD, IVSD, LVM, LVEDP, and LVSP were higher in the I/R, CNP-/- control, and CNP-/- groups than normal control group while LVEF, SV, CO, and ±dp/dtmax were lower. Compared with the normal control group, LDH, cTn I, CK-MB, TNF-α, and IL-6 were higher in the I/R, CNP-/- control and CNP-/- groups; pathological changes and myocardial infarction were observed in the I/R, CNP-/- control, and CNP-/- groups; expressions of apoptosis-related genes in those groups were higher; while CNP, NPRB, cGMP, and Bcl-2 expressions were decreased. We came to the conclusion that gene knockdown of CNP blocks the NPRB/cGMP signaling pathway, thereby aggravating myocardial I/R injury and causing ventricular remodeling in rats.

Generation of reporter hESCs by targeting EGFP at the CD144 locus to facilitate the endothelial differentiation.

Reporter embryonic stem cell (ESC) lines with tissue-specific reporter genes may contribute to optimizing the differentiation conditions in vitro as well as trafficking transplanted cells in vivo. To optimize and monitor endothelial cell (EC) differentiation specifically, here we targeted the enhanced green fluorescent protein (EGFP) reporter gene at the junction of 5'UTR and exon2 of the endothelial specific marker gene CD144 using TALENs in human ESCs (H9) to generate a EGFP-CD144-reporter ESC line. The reporter cells expressed EGFP and CD144 increasingly and specifically without unexpected effects during the EC differentiation. The EC differentiation protocol was optimized and applied to EC differentiation from hiPSCs, resulting in an efficient and simplified endothelial differentiation approach. Here we created our own optimized and robust protocol for EC differentiation of hESCs and hiPSCs by generating the lineage-specific site-specific integration reporter cell lines, showing great potential to be applied in the fields such as trafficking gene and cell fate in vivo in preclinical animal models.

Paired CRISPR/Cas9 Nickases Mediate Efficient Site-Specific Integration of F9 into rDNA Locus of Mouse ESCs.

Hemophilia B (HB) is an X-linked recessive bleeding disorder, caused by F9 gene deficiency. Gene therapy combined with the CRISPR/Cas9 technology offers a potential cure for hemophilia B. Now the Cas9 nickase (Cas9n) shows a great advantage in reducing off-target effect compared with wild-type Cas9. In this study, we found that in the multicopy ribosomal DNA (rDNA) locus, the homology directed recombination (HDR) efficiency induced by sgRNA-Cas9n was much higher than sgRNA-Cas9, meanwhile without off-target in six predicted sites. After co-transfection into mESCs with sgRNA-Cas9n and a non-viral rDNA targeting vector pMrnF9, harboring the homology donor template and the human F9 expression cassette, a recombination efficiency of 66.7% was achieved and all targeted clones were confirmed to be site-specific integration of F9 in the rDNA locus by PCR and southern blotting. Targeted mESCs retained the main pluripotent properties and were then differentiated into hepatic progenitor like cells (HPLCs) and mature hepatocytes, which were characterized by hepatic markers and functional assays. Importantly, the differentiated cells could transcribe exogenous F9 and secrete coagulation factor IX (FIX) proteins, suggesting active transcription and stable inheritance of transgenes in the rDNA locus. After intrasplenical transplantation in severe combined immune deficiency (SCID) mice, targeted HPLCs could survive and migrate from spleen to liver, resulting in secretion of exogenous FIX into blood. In summary, we demonstrate an efficient and site-specific gene targeting strategy in rDNA locus for stem cell-based gene therapy for hemophilia B.

Targeting of the human F8 at the multicopy rDNA locus in Hemophilia A patient-derived iPSCs using TALENickases.

Hemophilia A (HA) is a monogenic disease due to lack of the clotting factor VIII (FVIII). This deficiency may lead to spontaneous joint hemorrhages or life-threatening bleeding but there is no cure for HA until very recently. In this study, we derived induced pluripotent stem cells (iPSCs) from patients with severe HA and used transcription activator-like effector nickases (TALENickases) to target the factor VIII gene (F8) at the multicopy ribosomal DNA (rDNA) locus in HA-iPSCs, aiming to rescue the shortage of FVIII protein. The results revealed that more than one copy of the exogenous F8 could be integrated into the rDNA locus. Importantly, we detected exogenous F8 mRNA and FVIII protein in targeted HA-iPSCs. After they were differentiated into endothelial cells (ECs), the exogenous FVIII protein was still detectable. Thus, it is showed that the multicopy rDNA locus could be utilized as an effective target site in patient-derived iPSCs for gene therapy. This strategy provides a novel iPSCs-based therapeutic option for HA and other monogenic diseases.

Path analysis of influencing factors for maternal antenatal depression in the third trimester.

Maternal antenatal depression (AD) is a nonpsychotic depressive episode during pregnancy that can harm both the pregnant woman and the fetus. This study aimed to investigate the intrinsic interrelationships between AD and its influencing factors by constructing a path model. This survey-based cross-sectional study included 1071 pregnant women who underwent pregnancy examinations in three hospitals in Nantong City, China, between February and June 2023. General information and information regarding maternal AD, pregnancy stress, prenatal anxiety, social support, marital satisfaction, sleep quality, and resilience were collected. Multiple linear regression analysis using SPSS 25.0 was employed to determine the factors influencing pregnancy depression, and Amos25.0 was used to construct a structural equation model. AD incidence was 19.4% (208/1071). The independent risk factors affecting AD in pregnant women have been integrated into the established path analysis model. The model demonstrated a good fit (χ2/DF = 1.238, comparative fit index = 0.999, goodness-of-fit index = 0.998, normed fit index = 0.996, adjusted goodness-of-fit index = 0.990, incremental fit index = 0.999, and root mean square error of approximation = 0.015). While prenatal anxiety (0.230) and hyperthyroidism (0.048) only had direct effects on AD, mental resilience was the biggest factor affecting AD, followed by pregnancy stress, marital satisfaction, prenatal anxiety, sleep quality, social support, and hyperthyroidism. Improved mental resilience, social support, sleep quality, and marital satisfaction; reduced pregnancy stress and prenatal anxiety; and effective hyperthyroidism treatment might reduce AD. This study underscored the significance of delivering actionable strategies and tangible assistance to pregnant women to reduce AD.

A novel variation in DEPDC5 causing familial focal epilepsy with variable foci.

Background: Disheveled, EGL-10, and pleckstrin (DEP) domain-containing protein 5 (DEPDC5) is a component of GTPase-activating protein (GAP) activity toward the RAG complex 1 (GATOR1) protein, which is an inhibitor of the amino acid-sensing branch of the mammalian target of rapamycin complex 1 (mTORC1) pathway. GATOR1 complex variations were reported to correlate with familial focal epilepsy with variable foci (FFEVF). With the wide application of whole exome sequencing (WES), more and more variations in DEPDC5 were uncovered in FFEVF families. Methods: A family with a proband diagnosed with familial focal epilepsy with variable foci (FFEVF) was involved in this study. Whole exome sequencing (WES) was performed in the proband, and Sanger sequencing was used to confirm the variation carrying status of the family members. Mini-gene splicing assay was performed to validate the effect on the alternative splicing of the variation. Results: A novel variant, c.1217 + 2T>A, in DEPDC5 was identified by WES in the proband. This splicing variant that occurred at the 5' end of intron 17 was confirmed by mini-gene splicing assays, which impacted alternative splicing and led to the inclusion of an intron fragment. The analysis of the transcribed mRNA sequence indicates that the translation of the protein is terminated prematurely, which is very likely to result in the loss of function of the protein and lead to the occurrence of FFEVF. Conclusion: The results suggest that c.1217 + 2T>A variations in DEPDC5 might be the genetic etiology for FFEVF in this pedigree. This finding expands the genotype spectrum of FFEVF and provides new etiological information for FFEVF.

Plasma GPI and PGD are associated with vascular normalization and may serve as novel prognostic biomarkers for lung adenocarcinoma: Multi-omics and multi-dimensional analysis.

The aim of this study was to explore potential novel plasma protein biomarkers for lung adenocarcinoma (LUAD). A plasma proteomics analysis was carried out and candidate protein biomarkers were validated in 102 LUAD cases and 102 matched healthy controls. The same LUAD tumor tissues were detected to explore the correlation between the expression of candidate proteins in tissues and plasma and vascular normalization. A LUAD active metastasis mice model was constructed to explore the role of candidate proteins for lung metastasis. GPI and PGD were verified to be upregulated in plasma from LUAD patients, and the expression of GPI in tumor tissue was positively correlated with the expression of GPI in plasma and negatively correlated with the normalization of tumor blood vessels. Meanwhile, a negative correlation between the expression of GPI and PGD in plasma and tumor vascular normalization was discovered. In the LUAD active metastasis model, the lowest levels of vascular normalization and the highest expression of GPI and PGD were found in mice with lung metastases. This study found that GPI and PGD may be potential plasma biomarkers for LUAD, and monitoring those may infer the risk of metastasis and malignancy of the tumor. SIGNIFICANT: We identified GPI and PGD as potential novel diagnostic and prognostic biomarkers for LUAD. PGD and GPI can be used as diagnostic biomarkers in combination with other available strategies to assist in the screening and diagnosis of LUAD, and as prognostic biomarkers aid in predict the risk of tumor metastasis and malignancy in patients with LUAD.

Postpartum stress in the first 6 months after delivery: a longitudinal study in Nantong, China.

OBJECTIVES: The objective is to to explore the longitudinal change trajectories of postpartum stress and its related factors. DESIGN: A longitudinal study with follow-ups from 42 days to 6 months after delivery. SETTINGS AND PARTICIPANTS: A total of 406 postpartum women were recruited at baseline (42 days after delivery) from 6 hospitals in Nantong, Jiangsu Province, China, and followed up at 3 and 6 months. After the follow-ups, 358 postpartum women were retained for further analysis. METHODS: Postpartum stress was evaluated using the Maternal Postpartum Stress Scale (MPSS) at baseline (42 days) and 3 and 6 months after delivery. MPSS has three dimensions, such as: personal needs and fatigue, infant nurturing and body changes and sexuality. Postpartum depression and anxiety were measured using the Edinburgh Postnatal Depression Scale and the short-form Depression, Anxiety and Stress Scale, respectively. The MPSS scores were normalised using a rank-based inverse normal transformation. RESULTS: Postpartum stress decreased significantly after 3 months, and postpartum stress reduced further after 6 months. Additionally, the scores for all three dimensions reduced after 6 months, while infant nurturing reduced after both 3 and 6 months. Older age (ß=0.028, p=0.049), higher education level (ß=0.153, p=0.005) and higher body mass index (BMI) (ß=0.027, p=0.008) of the postpartum women were significantly associated with higher postpartum stress levels in corresponding dimensions at 42 days. Older age was also associated with higher postpartum stress at 3 (ß=0.030, p=0.033) and 6 months (ß=0.050, p<0.001) in the dimension of personal needs and fatigue. Postpartum stress levels were significantly higher in women with depression or anxiety symptoms. CONCLUSIONS: Postpartum stress continuously declined from 42 days to 6 months after delivery. Postpartum women with older age, higher education levels, higher BMI and anxiety or depression symptoms should be the target population for early intervention.

Gene Spectrum and Clinical Traits of Nine Patients With Oocyte Maturation Arrest.

Background: Oocyte maturation arrest is a disease that produces immature oocytes and cannot be mature after culturing in vitro, which leads to female primary infertility. We aimed to summarize nine representative patients in our center to retrospectively analyze the genetic variants and clinical characteristics of oocyte maturation arrest. Methods: This study examined and analyzed nine families with oocyte maturation arrest. Whole-exome sequencing (WES) of the probands was performed to detect the pathogenic variants. Sanger sequencing verified the WES findings in patients and available parents. ExAC database was used to search the variant frequency. The variants were assessed by pathogenicity and conservational property prediction analysis and according to the American College of Medical Genetics and Genomics (ACMG). Phenotypes of oocytes were evaluated by a light microscopy, and the phenotype-genotype correlation was also evaluated. Results: Nine pathogenic variants in five genes were detected in nine patients, of which three were novel variants, including PATL2 [c.1374A > G (p. Ile458Met)] and [1289-1291del TCC (p. Leu430del)] and ZP2 [c.1543C > T (p. Pro515Ser)]. Nine variants were predicted to be pathogenic, resulting in different types of oocyte maturation arrest and clinical phenotypes. Conclusion: Three novel pathogenic variants were identified, enabling the expansion of the gene variant spectrum. The related pathogenic mutations of the PATL2, TUBB8, and ZP1∼3 genes were highly suggestive of being causative of oocyte maturation arrest.

Restoration of SMN expression in mesenchymal stem cells derived from gene-targeted patient-specific iPSCs.

Spinal muscular atrophy (SMA) is primarily a neurodegenerative disease caused by the homozygous deletion of the survival motor neuron 1 (SMN1) gene, thereby reducing SMN protein expression. Mesenchymal stem cells (MSCs) have been implicated in the treatment of SMA. In the present study, we overexpressed exogenous SMN1 at the ribosomal DNA (rDNA) locus of induced pluripotent stem cells (iPSCs) generated from a SMA patient using an rDNA-targeting vector. The gene-targeted patient iPSCs differentiated into MSCs (SMN1-MSCs). A 2.1-fold higher expression level of SMN protein was detected in SMN1-MSCs than that detected in MSCs derived from patient iPSCs, and the results of the immunofluorescence analysis showed no difference in the quantity of SMN nuclear structures (gems) between SMN1-MSCs and MSCs derived from normal human iPSCs (h-MSCs). These findings provide a novel strategy for obtaining gene-targeted MSCs for potential clinical applications in autologous cell-based therapy.

Enhanced tumor growth inhibition by mesenchymal stem cells derived from iPSCs with targeted integration of interleukin24 into rDNA loci.

Induced pluripotent stem cells (iPSCs) are a promising source of mesenchymal stem cells (MSCs) for clinical applications. In this study, we transformed human iPSCs using a non-viral vector carrying the IL24 transgene pHrn-IL24. PCR and southern blotting confirmed IL24 integration into the rDNA loci in four of 68 iPSC clones. We then differentiated a high expressing IL24-iPSC clone into MSCs (IL24-iMSCs) that showed higher expression of IL24 in culture supernatants and in cell lysates than control iMSCs. IL24-iMSCs efficiently differentiated into osteoblasts, chondrocytes and adipocytes. Functionally, IL24-iMSCs induced in vitro apoptosis in B16-F10 melanoma cells more efficiently than control iMSCs when co-cultured in Transwell assays. In vivo tumor xenograft studies in mice demonstrated that IL24-iMSCs inhibited melanoma growth more than control iMSCs did. Immunofluorescence and histochemical analysis showed larger necrotic areas and cell nuclear aggregation in tumors with IL24-iMSCs than control iMSCs, indicating that IL24-iMSCs inhibited tumor growth by inducing apoptosis. These findings demonstrate efficient transformation of iPSCs through gene targeting with non-viral vectors into a rDNA locus. The ability of these genetically modified MSCs to inhibit in vivo melanoma growth is suggestive of the clinical potential of autologous cell therapy in cancer.