Therapeutic potential of urine-derived stem cells in renal regeneration following acute kidney injury:A comparative analysis with mesenchymal stem cells

BACKGROUND Acute kidney injury(AKI)is a common clinical syndrome with high morbidity and mortality rates.The use of pluripotent stem cells holds great promise for the treatment of AKI.Urine-derived stem cells(USCs)are a novel and versatile cell source in cell-based therapy and regenerative medicine that provide advantages of a noninvasive,simple,and low-cost approach and are induced with high multidifferentiation potential.Whether these cells could serve as a potential stem cell source for the treatment of AKI has not been determined.METHODS Stem cell markers with multidifferentiation potential were isolated from human amniotic fluid.AKI severe combined immune deficiency(SCID)mice models were induced by means of an intramuscular injection with glycerol.USCs isolated from human-voided urine were administered via tail veins.The functional changes in the kidney were assessed by the levels of blood urea nitrogen and serum creatinine.The histologic changes were evaluated by hematoxylin and eosin staining and transferase dUTP nick-end labeling staining.Meanwhile,we compared the regenerative potential of USCs with bone marrow-derived mesenchymal stem cells(MSCs).RESULTS Treatment with USCs significantly alleviated histological destruction and functional decline.The renal function was rapidly restored after intravenous injection of 5×105 human USCs into SCID mice with glycerol-induced AKI compared with injection of saline.Results from secretion assays conducted in vitro demonstrated that both stem cell varieties released a wide array of cytokines and growth factors.This suggests that a mixture of various mediators closely interacts with their biochemical functions.Two types of stem cells showed enhanced tubular cell prolif-eration and decreased tubular cell apoptosis,although USC treatment was not more effective than MSC treatment.We found that USC therapy significantly improved renal function and histological damage,inhibited inflammation and apoptosis processes in the kidney,and promoted tubular epithelial proliferation.CONCLUSION Our study demonstrated the potential of USCs for the treatment of AKI,representing a new clinical therapeutic strategy.

Analyses of oligodontia phenotypes and genetic etiologies

Oligodontia is the congenital absence of six or more teeth and comprises the more severe forms of tooth agenesis.Many genes have been implicated in the etiology of tooth agenesis,which is highly variable in its clinical presentation.The purpose of this study was to identify associations between genetic mutations and clinical features of oligodontia patients.An online systematic search of papers published from January 1992 to June 2021 identified 381 oligodontia cases meeting the eligibility criteria of causative gene mutation,phenotype description,and radiographic records.Additionally,ten families with oligodontia were recruited and their genetic etiologies were determined by whole-exome sequence analyses.We identified a novel mutation in WNT10A(c.99_105dup)and eight previously reported mutations in WNT10A(c.433 G>A;c.682 T>A;c.318 C>G;c.511.C>T;c.321 C>A),EDAR(c.581 C>T),and LRP6(c.1003 C>T,c.2747 G>T).Collectively,20 different causative genes were implicated among those 393 cases with oligodontia.For each causative gene,the mean number of missing teeth per case and the frequency of teeth missing at each position were calculated.Genotype–phenotype correlation analysis indicated that molars agenesis is more likely linked to PAX9 mutations,mandibular first premolar agenesis is least associated with PAX9 mutations.

The varied rate of response to dietary intervention in autistic children

Exorphins from casein and gluten have been found by HPLC and mass spectroscopy with fragmentation pattern in quickly frozen urine. Removing the proteins that contain these peptides, by dietary intervention has been tried with behavioral effects. We wanted to know how fast such changes take place. Method: Parents and caregivers filled out ATEC scores (Autism treatment evaluation checklist) over time so that changes in scores could be registered. Results: In this group of children who all responded to diet the time required for a positive effect was months rather than weeks. Conclusion: Short term interventions are probably a waste of time and money, and at least 3 - 6 months trials seem to be necessary.

Exorphins in urine from schizoaffective psychotics

Hyperpeptiduria and opioid excess have been reported in schizophrenia. According to Prof. Dr. L. Lindstrom, Sweden opioids may explain the patho-physiology of this syndrome. Therefore it is critical to elucidate the presence and nature of opioids in schizophrenia and diagnostic sub groups. First morning urine from untreated schizoaffective patients (ICD-10: F 25.1) was separated on HPLC and peaks that elute where different opioid standards appear, freeze dried, re-dissolved in methanol/water (50/50) and 10mM formic acid. Mass spectrometry and MS/MS or fragmentation mass spectrometry was performed. We found fragmentation pattern of beta-casomorphin 1-3 and 1-4 (bovine) identical to synthetic standards from Bachem. The aggregation tendency of peptides was much in evidence. The reported exorphins were found in the urine from 8 of 12 untreated schizoaffective patients.

N-Acetylcysteine Amide(NACA) Reduces Cell Death after Oxidative Stress in a Porcine Embryonic Kidney Cell Line

Introduction: Oxidative stress may have detrimental effects on different structures of the cells, such as the DNA. Recently, we have published a study demonstrating that N-Acetylcysteine amide (NACA) displayed anti-inflammatory properties on the brain after exposure to oxidative stress in an established neonatal piglet model, imitating perinatal asphyxia. As different clinical studies have shown an association between the severity of hypoxic-ischemic encephalopathy and damage of the kidneys, we investigated a possible protective effect of NACA against H2O2-induced oxidative stress using a porcine epithelial-like embryonic kidney cell line (EFN-R). Objective: To investigate a potential protective effect of NACA on cells of a porcine embryonic kidney cell line exposed to H2O2. Methods: We subjected the cells to different concentrations of H2O2 for variable time periods, seeking the optimal dose-response for the experiments. Based on the results of these investigations, we exposed the cells to 100 μMol of H2O2 and/or 750 μM of NACA for 24 hours. Some of the cells would receive NACA either one hour before or one hour after exposure to H2O2. Results: The viability of the investigated EFN-R cells revealed that both, the group treated with NACA before exposure to H2O2 and the group treated with NACA after exposure to H2O2, exhibited significantly higher cell viability compared to the H2O2 group (p < 0.001 and p < 0.01, respectively). Discussion: The increased viability of the cells may indicate that NACA could play an important role in reducing oxidative stress. Taking the results from our previous study into consideration, our findings may strengthen the theory that NACA may have organ protective properties for neonates exposed to oxidative stress.

PTEN-induced putative kinase 1 regulates mitochondrial quality control and is essential for the maturation of human induced pluripotent stem cell-derived cardiomyocytes

Human induced pluripotent stem cell-derived cardiomyocytes(hiPSC-CMs)have attracted attention in the field of regenerative medicine due to their potential ability to repair damaged hearts.However,the immature phenotype of these cells limits their clinical application.Cardiomyocyte maturation is accompanied by changes in mitochondrial quality.PTEN-induced putative kinase 1(PINK1)has been linked to mitochondrial quality control.However,whether the changes in mitochondrial quality in hiPSC-CMs are associated with PINK1,and the impact of PINK1 on hiPSC-CMs development are not clear.In this study,we found that knockdown of PINK1 in hiPSC-CMs resulted in mitochondrial fragmentation and impaired mitochondrial functions such as mitophagy and mitochondrial biogenesis.PINK1 deletion also inhibited the maturation of hiPSC-CMs,reverting them to a naive structural and functional state.We found that restoring the mitochondrial structure did not completely rescue the mitochondrial dysfunction caused by PINK1 deletion,while activation of PINK1 kinase activity using kinetin promoted mitochondrial fusion,increased the mitochondrial membrane potential and ATP production,and maintained the development and maturation of hiPSC-CMs.In conclusion,PINK1 regulates the mitochondrial structure and function of hiPSC-CMs,and is essential for the maturation of hiPSC-CMs.

Dedifferentiated human umbilical cord mesenchymal stem cell reprogramming of endogenous hSDF-1a expression participates in neural restoration in hypoxic-ischemic brain damagerats

The transplantation of human umbilical cord mesenchymal stem cells(hUC-MSCs)can promote hypoxic-ischemic brain damage(HIBD)nerve repair,but finding suitable seed cells to optimize transplantation and improve treatment efficiency is an urgent problem to be solved.In this study,we induced hUC-MSCs into dedifferentiated hUC-MSCs(De-hUC-MSCs),and the morphology,stem cell surface markers,proliferation and tri-directional differentiation ability of the De-hUC-MSCs and hUC-MSCs were detected.A whole-gene chip was utilized for genome cluster,gene ontology and KEGG pathway analyses of differentially expressed genes.De-hUC-MSCs were transplanted into HIBD rats,and behavioral experiments and immunofluorescence assays were used to assess the therapeutic effect.A lentivirus vector for human stromal cell-derived factor-1(hSDF-1a)was constructed,and the role of hSDF-1a in the neuroprotective effect and mechanism of De-hUC-MSCs was verified.De-hUC-MSCs displayed similar cell morphology,stem cell surface marker expression,cell proliferation and even three-dimensional differentiation ability as hUC-MSCs but exhibited greater treatment potential in vivo.The reprogramming mechanism of hSDF-1a participated in the dedifferentiation process.By successfully constructing a stable hSDF-1a cell line,we found that De-hUC-MSCs might participate in nerve repair through the hSDF-1a/CXCR4/PI3K/Akt pathway.De-hUC-MSCs reprogramming of endogenous hSDF-1a expression may mediate the hSDF-1a/CXCR4/PI3K/Akt pathway involved in nerve repair in HIBD rats.

PGC-1α promotes mitochondrial respiration and biogenesis during the differentiation of hiPSCs into cardiomyocytes

Although it is widely accepted that human induced pluripotent stem cell-derived cardiomyocytes(hiPSC-CMs)are readily available,robustly reproducible,and physiologically appropriate human cells for clinical applications and research in the cardiovascular field,hiPSC-CMs cultured in vitro retain an immature metabolic phenotype that limits their application,and little is known about the underlying molecular mechanism controlling mitochondrial metabolic maturation during human induced pluripotent stem cells(hiPSCs)differentiation into cardiomyocytes.In this study,we found that peroxisome proliferator-activated receptor g coactivator-1α(PGC-1α)played an important role in inducing mitochondrial biogenesis and establishing oxidative phosphorylation(OXPHOS)during the cardiac differentiation of hiPSCs.Knocking down PGC-1α by siRNA impaired mitochondrial respiration,while upregulating PGC-1α by ZLN005 promoted mitochondrial biosynthesis and function by regulating the expression of downstream genes involved in mitochondrial dynamics and oxidative metabolism in hiPSCCMs.Furthermore,we found that estrogen-related receptor a(ERRa)was required for the induction of PGC-1α stimulatory effects in hiPSC-CMs.These findings provide key insights into the molecular control of mitochondrial metabolism during cardiac differentiation and may be used to generate more metabolically mature cardiomyocytes for application.

欧洲新生儿呼吸窘迫综合征防治共识指南：2016版

新生儿呼吸窘迫综合征（RDS）是早产儿的重要疾病,尽管治疗手段不断成熟,低龄早产儿存活率逐渐增高,但支气管肺发育不良（BPD）发病率也随之升高,其中部分原因是减少了出生后激素的使用.2006年以来,来自欧洲许多国家的新生儿专家每3年一次回顾最新文献,就RDS或有RDS风险早产儿的防治达成共识,立志于改善欧洲新生儿的预后.欧洲RDS防治指南2007年开始发布,2010年和2013年进行了更新,期间获得了欧洲围产医学学会的大力支持.RDS是由于肺表面活性物质（PS）缺乏导致生后早期出现呼吸窘迫,典型临床表现随着防治手段的改进发生了巨大变化.

欧洲新生儿呼吸窘迫综合征防治指南-2010版

呼吸窘迫综合征（RDS）是由于肺表面活性物质（PS）缺乏及肺结构发育不成熟所致，多见于早产儿，自然病程为生后当时或很快发病，并在生后2d内进行性恶化，如不及时治疗，因进行性缺氧和呼吸衰竭而死亡，存活者，在生后2—4d病情开始改善。胎龄越小，RDS发生率越高，2006年EuroNeoStat的数据显示：胎龄23～25周的早产儿RDS发生率为91％，

欧洲早产儿呼吸窘迫综合征防治共识指南（2013版）

本次指南更新包括了自2010年以来Cochrane系统评价和医学文献中的最新证据，并采纳了一种新的证据评估分级系统（GRADE）。以往关于早期肺表面活性物质和持续气道正压通气治疗的推荐目前拥有了更坚定的循证依据。对产房复苏、稳定生命体征等内容进行了较大扩充。生命体征稳定后的氧疗目前仍存在一定的争议，但是在获得更多的证据之前，血氧饱和度的目标范围不应低于90％，支持治疗对于超早早产儿仍然极为重要。指南对其他注意事项进行了缩减。

Novel biallelic TRNT1 mutations lead to atypical SIFD and multiple immune defects

Mutations in the gene encoding transfer RNA(tRNA)nucleotidyltransferase,CCAadding 1(TRNT1),an enzyme essential for the synthesis of the 30-terminal CCA sequence in tRNA molecules,are associated with a rare syndrome of congenital sideroblastic anemia,B cell immunodeficiency,periodic fevers,and developmental delay(SIFD).Clinical manifestations and immunological phenotypes were assessed in a Chinese patient with novel compound heterozygous mutations in TRNT1.The patient required multiple hospitalizations starting at the age of 2 years for recurrent fevers without an infective cause.During the febrile episode,the patient was found to have microcytic hypochromic anemia,B cell lymphopenia,and hypogammaglobulinemia.Targeted gene sequencing identified novel compound heterozygous mutations in the TRNT1 gene(c.525delT,p.Leu176X;c.938T>C,p.Leu313Ser).Immunophenotyping revealed increased CD8^+T cells,CD4^+ terminally differentiated effector memory helper T lymphocytes(CD4 TEMRA),and CD4^+ effector memory lymphocytes(CD4 EM).Analysis of CD4^+T subsets identified decreased T follicular helper cells(Tfh)with a biased phenotype to Th2-like cells.The patient also showed a lower percentage of switched memory B(smB)cells.Additionally,defects in the cytotoxicity of the patient’s NK andγτT cells were shown by CD107alpha expression.In conclusion,TRNT1 mutations may lead to multiple immune abnormality especially humoral and cytotoxicity defects,which indicate that SIFD is not only suffered‘Predominantly antibody deficiencies’in IUIS classification system,and further studies are needed to understand the pathogenesis of immunodeficiency in these patients.

Bone morphogenetic protein 4 (BMP4) promotes hepatic glycogen accumulation and reduces glucose level in hepatocytes through mTORC2 signaling pathway

Liver is an important organ for regulating glucose and lipid metabolism.Recent studies have shown that bone morphogenetic proteins(BMPs)may play important roles in regu-lating glucose and lipid metabolism.Inour previous studies,we demonstrated that BMP4 signif-icantly inhibits hepatic steatosis and lowers serum triglycerides,playing a protective role against the progression of non-alcoholic fatty liver disease(NAFLD).However,the direct impact of BMP4 on hepatic glucose metabolism is poorly understood.Here,we investigated the regulatory roles of BMP4 in hepatic glucose metabolism.Through a comprehensive analysis of the 14 types of BMPs,we found that BMP4 was one of the most potent BMPs in promoting hepatic glycogen accumulation,reducing the level of glucose in hepatocytes and effecting the expression of genes related to glucose metabolism.Mechanistically,we demonstrated that BMP4 reduced the hepatic glucose lewels through the activation of mTORC2 signaling pathway in vitro and in wivo.Collectively,our findings strongly suggest that BMP4 may play an essential role in regulating hepatic glucose metabolism.This knowledge should aid us to understand the molecular pathogenesis of NAFLD,and may lead to the development of novel therapeutics by exploiting the inhibitory effects of BMPs on hepatic glucose and lipid metabolism.

Publishing clinical prActice GuidelinEs(PAGE):Recommendations from Editors and Reviewers

Transparency Ecosystemfor Research and Journals inMedicine(TERM)Working Group summarized the essential recommendations that should be considered to review and publish a high-quality guideline.These recommendations fromeditors and reviewers included the 10 components of essential requirements:systematic review of existing relevant guidelines,guideline registration,guideline protocol,stakeholders,conflicts of interest,clinical questions,systematic reviews,recommendation consensus,guideline reporting,and external review.TERMWorking Group abbreviates them as PAGE(essential requirements for Publishing clinical prActice GuidelinEs),recommends guideline authors,editors,and peer reviewers use them for high-quality guidelines.

Publishing clinical prActice GuidelinEs(PAGE):Recommendations from editors and reviewers

Transparency Ecosystem for Research and Journals in Medicine(TERM)working group summarized the essential recommendations that should be considered to review and publish a high-quality guideline.These recommendations from editors and reviewers included 10 components of essential requirements:systematic review of existing relevant guidelines,guideline registration,guideline protocol,stakeholders,conflicts of interest,clinical questions,systematic reviews,recommendation consensus,guideline reporting and external review.TERM working group abbreviates them as PAGE(essential requirements for Publishing clinical prActice GuidelinEs),and recommends guideline authors,editors,and peer reviewers to use them for high-quality guidelines.