Human umbilical cord mesenchymal stem cells reverse depression in rats induced by chronic unpredictable mild stress combined with lipopolysaccharide.

BACKGROUND: Inflammation and oxidative stress are considered crucial to the pathogenesis of depression. Rat models of depression can be created by combined treatments of chronic unpredictable mild stress (CUMS) and lipopolysaccharide (LPS). Behaviors associated with depression could be improved by treatment with mesenchymal stem cells (MSCs) owing to immunomodulatory functions of the cells. Therapeutic potentials of the MSCs to reverse pro-inflammatory cytokines, proteins, and metabolites were identified by transcriptomic, proteomic, and metabolomic analysis, respectively. METHODS: A depression model was established in male SD rats by 2 weeks of CUMS combined with LPS. The models were verified by behavioral tests, namely SPT, OFT, EPM, and qRT-PCR for pro-inflammatory cytokines. Such depressed rats were administered human umbilical cord MSCs (hUC-MSCs) via the tail vein once a week for 2 and 4 weeks. The homing capacity was confirmed by detection of the fluorescent dye on day 7 after the hUC-MSCs were labeled with CM-Dil and administered. The expression of GFAP in astrocytes serves as a biomarker of CNS disorders and IBA1 in microglia serves as a marker of microglia activation were detected by immunohistochemistry at 2 and 4 weeks after final administration of hUC-MSCs. At the same time, transcriptomics of rat hippocampal tissue, proteomic and metabolomic analysis of the serum from the normal, depressed, and treated rats were also compared. RESULTS: Reliable models of rat depression were successfully induced by treatments of CUMS combined with LPS. Rat depression behaviors, pro-inflammatory cytokines, and morphological disorders of the hippocampus associated with depression were reversed in 4 weeks by hUC-MSC treatment. hUC-MSCs could reach the hippocampus CA1 region through the blood circulation on day 7 after administration owing to the disruption of blood brain barrier (BBB) by microglial activation from depression. Differentiations of whole-genome expression, protein, and metabolite profiles between the normal and depression-modeled rats, which were analyzed by transcriptomic, proteomics, and metabolomics, further verified the high association with depression behaviors. CONCLUSIONS: Rat depression can be reversed or recovered by treatment with hUC-MSCs.

A grayscale compression method to segment bone structures for 2D-3D registration of setup images in non-coplanar radiotherapy.

Purpose. To evaluate the performance of an automated 2D-3D bone registration algorithm incorporating a grayscale compression method for quantifying patient position errors in non-coplanar radiotherapy.Methods. An automated 2D-3D registration incorporating a grayscale compression method to segment bone structures was proposed. Portal images containing only bone structures (Portalbone) and digitally reconstructed radiographs containing only bone structures (DRRbone) were used for registration. First, the portal image was filtered by a high-pass finite impulse response (FIR) filter. Then the grayscale range of the filtered portal image was compressed. Thresholds were determined based on the difference in gray values of bone structures in the filtered and compressed portal image to obtainPortalbone.Another threshold was applied to generateDRRbonewhen the CT image uses the ray-casting algorithm to generate DRR images. The compression performance was assessed by registering theDRRbonewith thePortalboneobtained by compressing the portal image into various grayscale ranges. The proposed registration method was quantitatively and visually validated using (1) a CT image of an anthropomorphic head phantom and its portal images obtained in different poses and (2) CT images and pre-treatment portal images of 20 patients treated with non-coplanar radiotherapy.Results. Mean absolute registration errors for the best compression grayscale range test were 0.642 mm, 0.574 mm, and 0.643 mm, with calculation times of 50.6 min, 42.2 min, and 49.6 min for grayscale ranges of 0-127, 0-63 and 0-31, respectively. For the accuracy validation (1), the mean absolute registration errors for couch angles 0°, 45°, 90°, 270°, and 315° were 0.694 mm, 0.839 mm, 0.726 mm, 0.833 mm, and 0.873 mm, respectively. Among the six transformation parameters, the translation error in the vertical direction contributed the most to the registration errors. Visual inspection of the patient registration results revealed success in every instance.Conclusions. The implemented grayscale compression method successfully enhances and segments bone structures in portal images, allowing for accurate determination of patient setup errors in non-coplanar radiotherapy.

Translocation of telomerase reverse transcriptase coincided with ATP release in postnatal cochlear supporting cells.

The spontaneous bursts of electrical activity in the developing auditory system are derived from the periodic release of adenosine triphosphate (ATP) by supporting cells in the Kölliker's organ. However, the mechanisms responsible for initiating spontaneous ATP release have not been determined. Our previous study revealed that telomerase reverse transcriptase (TERT) is expressed in the basilar membrane during the first postnatal week. Its role in cochlear development remains unclear. In this study, we investigated the expression and role of TERT in postnatal cochlea supporting cells. Our results revealed that in postnatal cochlear Kölliker's organ supporting cells, TERT shifts from the nucleus into the cytoplasm over time. We found that the TERT translocation tendency in postnatal cochlear supporting cells in vitro coincided with that observed in vivo. Further analysis showed that TERT in the cytoplasm was mainly located in mitochondria in the absence of oxidative stress or apoptosis, suggesting that TERT in mitochondria plays roles other than antioxidant or anti-apoptotic functions. We observed increased ATP synthesis, release and activation of purine signaling systems in supporting cells during the first 10 postnatal days. The phenomenon that TERT translocation coincided with changes in ATP synthesis, release and activation of the purine signaling system in postnatal cochlear supporting cells suggested that TERT may be involved in regulating ATP release and activation of the purine signaling system. Our study provides a new research direction for exploring the spontaneous electrical activity of the cochlea during the early postnatal period.

A Single-Cell Atlas of an Early Mongolian Sheep Embryo.

Cell types have been established during organogenesis based on early mouse embryos. However, our understanding of cell types and molecular mechanisms in the early embryo development of Mongolian sheep has been hampered. This study presents the first comprehensive single-cell transcriptomic characterization at E16 in Ujumqin sheep and Hulunbuir short-tailed sheep. Thirteen major cell types were identified at E16 in Ujumqin sheep, and eight major cell types were identified at E16 in Hulunbuir short-tailed sheep. Function enrichment analysis showed that several pathways were significantly enriched in the TGF-beta signaling pathway, the Hippo signaling pathway, the platelet activation pathway, the riboflavin metabolism pathway, the Wnt signaling pathway, regulation of the actin cytoskeleton, and the insulin signaling pathway in the notochord cluster. Glutathione metabolism, glyoxylate, and dicarboxylate metabolism, the citrate cycle, thyroid hormone synthesis, pyruvate metabolism, cysteine and methionine metabolism, thermogenesis, and the VEGF signaling pathway were significantly enriched in the spinal cord cluster. Steroid biosynthesis, riboflavin metabolism, the cell cycle, the Hippo signaling pathway, the Hedgehog signaling pathway, the FoxO signaling pathway, the JAK-STAT signaling pathway, and the Wnt signaling pathway were significantly enriched in the paraxial mesoderm cluster. The notochord cluster, spinal cord cluster, and paraxial mesoderm cluster were found to be highly associated with tail development. Pseudo-time analysis demonstrated that the mesenchyme can translate to the notochord in Ujumqin sheep. Molecular assays revealed that the Hippo signaling pathway was enriched in Ujumqin sheep. This comprehensive single-cell map revealed previously unrecognized signaling pathways that will further our understanding of the mechanism of short-tailed sheep formation.

Reprogramming efficiency and pluripotency of mule iPSCs over its parents†.

The mule is the interspecific hybrid of horse and donkey and has hybrid vigor in muscular endurance, disease resistance, and longevity over its parents. Here, we examined adult fibroblasts of mule (MAFs) compared with the cells from their parents (donkey adult fibroblasts and horse adult fibroblasts) (each species has repeated three independent individuals) in proliferation, apoptosis, and glycolysis and found significant differences. We subsequently derived mule, donkey, and horse doxycycline (Dox)-independent induced pluripotent stem cells (miPSCs, diPSCs, and hiPSCs) from three independent individuals of each species and found that the reprogramming efficiency of MAFs was significantly higher than that of cells of donkey and horse. miPSCs, diPSCs, and hiPSCs all expressed the high levels of crucial endogenous pluripotency genes such as POU class 5 homeobox 1 (POU5F1, OCT4), SRY-box 2 (SOX2), and Nanog homeobox (NANOG) and propagated robustly in single-cell passaging. miPSCs exhibited faster proliferation and higher pluripotency and differentiation than diPSCs and hiPSCs, which were reflected in co-cultures and separate-cultures, teratoma formation, and chimera contribution. The establishment of miPSCs provides a unique research material for the investigation of "heterosis" and perhaps is more significant to study hybrid gamete formation.

Analyzing the Role of Septin9 Gene Methylation in the Diagnosis and Treatment of Primary Liver Cancer in the Elderly.

Context: Because the early symptoms of primary hepatocellular carcinoma (PHC) aren't significant, it's difficult to diagnose it by routine inspection clinically, and if the lesion's diameter is small, less than 2.0 cm, false negatives can occur in pathological examinations. Researchers need to actively search for more diagnostic methods. Objective: The study intended to detect and analyze the value of plasma Septin9 gene methylation for the diagnosis and therapeutic monitoring of PHC in older adults. Design: The research team performed a prospective controlled study. Setting: The study took place at the First Hospital of Qiqihar, an Affiliated Qiqihar Hospital at Southern Medical University in Qiqihar, China. Participants: Participants were 32 patients with PHC and 28 with cholangiocarcinoma (CCA) who had been admitted to the hospital between January 2021 and July 2022 and 40 healthy individuals. Groups: The research team divided participants into three groups: (1) patients with PHC, the PHC group; (2) patients with CCA, the CCA group; and (3) healthy individuals, the control group. Outcome Measures: The research team: (1) determined the positive expression rate of Septin9 gene methylation; (2) measured liver function indicators-alanine aminotransferase (ALT), aspartate aminotransferase (AST), serum total bilirubin (TBIL), direct bilirubin (DBIL), alkaline phosphatase (ALP), γ-glutamyl transpeptidase (GGT), albumin (ALB); and (3) measured tumor markers-alpha-fetoprotein (AFP), carbohydrate antigen (CA) 199, CA125, and CA153. The team also: (1) established a binary logistic regression model based on levels of GGT and plasma Septin9 gene methylation to analyze risk factors and diagnosis accuracy, (2) created a receiver operating characteristic (ROC) curve to analyze diagnostic values; and (3) during followup, analyzed the negative conversion rate of Septin9 gene methylation in participants. Results: The positive expression rate of Septin9 methylation in the PHC group was significantly lower than that that of the CCA group and significantly higher than that of the control group (P < .05). The PHC group's ALT, AST, TBIL, DBIL, ALP, and GGT were significantly higher than those of the control group but significantly lower than those of the CCA group (all P < .05). PHC group's ALB was significantly lower than that of the control group (P < .05). The PHC group's AFP, CA199, and CA125 were significantly higher than those of the control group, and the PHC group's CA199 and CA125 were significantly lower than those in the CCA group (all P < .05). The positive expression of Septin9 gene methylation and the high expression of GGT were risk factors for PHC (OR>1, P < .05). The AUC of the Septin9 gene methylation, the GGT level, and the combined detection of both variables (all AUC > 0.70), suggests that the variables have a diagnostic value in the detection of PHC, with the combined detection having the highest value. The negative conversion rate after surgery of Septin9 gene methylation was 87.10%, for 27 out of 31 participants in the PHC and CCA groups (χ2 = 29.405, P < .001). Conclusion: Plasma Septin9 gene methylation is a sensitive molecular marker for the diagnosis and therapeutic monitoring of older adults with PHC, and combined with the serum GGT level, has a high diagnostic efficiency, which may reflect the treatment status of patients.

[The role of triangular space of ear canal-parotid-mastoid in the surgery for first branchial cleft anomalies].

Objective:To explore the role of triangular space of ear canal-parotid-mastoid in the operation of the first branchial cleft deformity. Methods:The clinical features and intraoperative characteristics of 25 cases with first branchial cleft anomalies who underwent surgery from September 2011 to September 2019 were analyzed, and the role of the triangular space of ear canal-parotid-mastoid in the surgery was explored. Results:Following dissecting and lesions removel of the triangular space of ear canal-parotid-mastoid, all the lesions were resected completely. Eighteen cases had fistula in the floor wall of ear canal, seven cases had duplicated of external auditory canal in the inferior of the floor wall. The recurrent cases were all attributable to the residual lesions in the triangular space. There was no recurrence, salivary leakage or stenosis of external canal. One case suffered from HB2 level facial paralysis. Conclusion:Surgery is the optimal treatment for first branchial cleft anomalies. Following the active dissection of the ear canal-parotid gland-mastoid space and depending on the microscopic operation, the deep lesions would be exposed clearly and the facial nerve could be marked and protected. Cleaning this triangle space can lead to completely lesion removal, avoid facial paralysis, salivation and recurrence.

Au3+ -Functionalized UiO-67 Metal-Organic Framework Nanoparticles: O2•- and •OH Generating Nanozymes and Their Antibacterial Functions.

The synthesis and characterization of Au3+ -modified UiO-67 metal-organic framework nanoparticles, Au3+ -NMOFs, are described. The Au3+ -NMOFs reveal dual oxidase-like and peroxidase-like activities and act as an active catalyst for the catalyzed generation of O2•- under aerobic conditions or •OH in the presence of H2 O2 . The two reactive oxygen species (ROS) agents O2•- and •OH are cooperatively formed by Au3+ -NMOFs under aerobic conditions, and in the presence of H2 O2. The Au3+ -NMOFs are applied as an effective catalyst for the generation ROS agents for antibacterial and wound healing applications. Effective antibacterial cell death and inhibition of cell proliferation of Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli) bacterial colonies are demonstrated in the presence of the Au3+ -NMOFs. In addition, in vivo experiments demonstrate effective wound healing of mice wounds infected by S. aureus, treated by the Au3+ -NMOFs.

Global Transcriptional Analyses of the Wnt-Induced Development of Neural Stem Cells from Human Pluripotent Stem Cells.

The differentiation of human pluripotent stem cells (hPSCs) to neural stem cells (NSCs) is the key initial event in neurogenesis and is thought to be dependent on the family of Wnt growth factors, their receptors and signaling proteins. The delineation of the transcriptional pathways that mediate Wnt-induced hPSCs to NSCs differentiation is vital for understanding the global genomic mechanisms of the development of NSCs and, potentially, the creation of new protocols in regenerative medicine. To understand the genomic mechanism of Wnt signaling during NSCs development, we treated hPSCs with Wnt activator (CHIR-99021) and leukemia inhibitory factor (LIF) in a chemically defined medium (N2B27) to induce NSCs, referred to as CLNSCs. The CLNSCs were subcultured for more than 40 passages in vitro; were positive for AP staining; expressed neural progenitor markers such as NESTIN, PAX6, SOX2, and SOX1; and were able to differentiate into three neural lineage cells: neurons, astrocytes, and oligodendrocytes in vitro. Our transcriptome analyses revealed that the Wnt and Hedgehog signaling pathways regulate hPSCs cell fate decisions for neural lineages and maintain the self-renewal of CLNSCs. One interesting network could be the deregulation of the Wnt/ß-catenin signaling pathway in CLNSCs via the downregulation of c-MYC, which may promote exit from pluripotency and neural differentiation. The Wnt-induced spinal markers HOXA1-4, HOXA7, HOXB1-4, and HOXC4 were increased, however, the brain markers FOXG1 and OTX2, were absent in the CLNSCs, indicating that CLNSCs have partial spinal cord properties. Finally, a CLNSC simple culture condition, when applied to hPSCs, supports the generation of NSCs, and provides a new and efficient cell model with which to untangle the mechanisms during neurogenesis.

Msi1 promotes breast cancer metastasis by regulating invadopodia-mediated extracellular matrix degradation via the Timp3-Mmp9 pathway.

Metastasis is the main cause of death in breast cancer patients. The initial step of metastasis is invadopodia-mediated extracellular matrix (ECM) degradation, which enables local breast tumor cells to invade surrounding tissues. However, the molecular mechanism underlying invadopodia-mediated metastasis remains largely unknown. Here we found that the RNA-binding protein Musashi1 (Msi1) exhibited elevated expression in invasive breast tumors and promoted lung metastasis of mammary cancer cells. Suppression of Msi1 reduced invadopodia formation in mammary cancer cells. Furthermore, Msi1 deficiency decreased the expression and activity of Mmp9, an important enzyme in ECM degradation. Mechanistically, Msi1 directly suppressed Timp3, an endogenous inhibitor of Mmp9. In clinical breast cancer specimens, TIMP3 and MSI1 levels were significantly inversely correlated both in normal breast tissue and breast cancer tissues and associated with overall survival in breast cancer patients. Taken together, our findings demonstrate that the MSI1-TIMP3-MMP9 cascade is critical for invadopodia-mediated onset of metastasis in breast cancer, providing novel insights into a promising therapeutic strategy for breast cancer metastasis.

[Comparative analysis of the hearing of three type reconstruction material in type â ¡ tympanoplasty of 286 cases].

Objective:To compare the effect on hearing of different reconstruction material in type Ⅱ tympanoplasty. Methods:Retrospectively analysis of 286 patients who accepted type Ⅱ tympanoplasty. The air-bone gap of 0.5, 1, 2, 4 kHz was analyzed before and after operation. We compared the hearing change and the complications between each group. Results:In incus group, the manubrium mallei and the head of stapes were connected with shaped incus, PORP group were implanted with PORP during operation, and cartilage group used auricular cartilage to cover the head of stapes. There was no significant difference in 4 kHz air-bone gap（ABG） between the cartilage group and PORP group either before or after the operation （P>0.05）. Air-bone gap of 0.5, 1, 2, 4 kHz of the incus group, and the 0.5, 1, 2 kHz of the cartilage and PORP group were significantly reducedafter the operation（P<0.05）. One patient got severe sensorineural hearing loss in incus group after the operation. The high frequency of bone conduction decreased in 1 patient（2, 4 kHz）.In the incus group, 3 patients had temporary facial paralysis after operation. Incus and cartilage group each have 1 patient with dizziness after the operation. Incus, cartilage and PORP group had 5, 3 and 11 patients with perforation again respectively. There was extrusion occurred in 1 patient of PORP group. Conclusion:Self incus, cartilage and PORP can be used in typeⅡ tympanoplasty, the effect of hearing reconstruction is similar. The first two are more economical, PORP implantation has the lowest technical difficulty and the most widely application, but there is a certain risk of extrusion .

[Evaluation of clinical application of new remote radar monitoring equipment for obstructive sleep apnea].

Objective:The result consistency between the new remote radar monitoring equipment and PSG detection is compared. Methods:The monitoring results of 71 patients were randomly selected to observe and compare the difference of monitoring data of the same patient with two kinds of detection equipment. Results:Using AHI≥5 as the diagnostic criteria, compared with PSG monitoring results, the sensitivity, specificity and Youden index of the new remote radar monitoring equipment were 96.2%, 89.5% and 0.857（P<0.01） respectively. Compared with PSG monitoring results, when LSaO2≤90% in PSG monitoring, the sensitivity, specificity and Youden index of the new remote radar monitoring equipment were 94.3%, 88.9% and 0.832（P<0.01） respectively. Conclusion: There the new remote radar monitoring device can be used to monitor OSA.

Author Correction: MiR-31 promotes mammary stem cell expansion and breast tumorigenesis by suppressing Wnt signaling antagonists.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

[Analysis of comorbid psychiatric disorders in patients with chronic otitis media associated tinnitus].

Objective:To analyze the distribution of comorbid psychiatric disorders in patients with chronic otitis media associated tinnitus. Method:The data of patients with chronic otitis media associated tinnitus who accepted surgical treatments from July 2017 to September 2018 were retrospectively analyzed. All patients accepted pure tone audiometry and acoustic conductance examination and were requested to fill the tinnitus history questionnaire, THI, TEQ, SAS, SDS and PSQI scales before operation. When the SAS or/and SDS score ≥50 the patient was judged as having comorbid psychiatric disorders. When the PSQI score>6 the patient was judged as having comorbid sleep disorder, and then all the results were analyzed. Result:Sixty-two patients were included in the study, 43 cases were diagnosed as chronic suppurative otitis media, and 19 cases were diagnosed as middle ear cholesteatoma. The average course of chronic otitis media or middle ear cholesteatoma was（14.38±14.06） years, while the average course of tinnitus was（8.39±11.69） years. There were 32 cases with mild to moderate tinnitus（gradeⅠ-Ⅱ）（51.61%） and 30 cases with moderate to severe tinnitus（grade Ⅲ-Ⅴ）（48.39%）. Before operation, there were 4 cases（6.45%） with normal hearing, 38 cases（61.29%） with conductive hearing loss, and 20 cases（32.36%） with mixed hearing loss. There was no significant difference in tinnitus severity between different hearing loss degrees and types（P>0.05）. The average SAS score was 45.10±11.61, and the average SDS score was 43.48±14.67, both higher than the normal modulus（30 points）, among which 27 cases（44.00%） comorbid psychiatric disorders. The THI score in patients with comorbid psychiatric disorders（57.85±21.1） was significantly higher than that in patients without comorbid psychiatric disorders（29.2±17.39）（P<0.05）. The PSQI score in patients with comorbid psychiatric disorders（8.86±3.47） was significantly higher than that of those without comorbid psychiatric disorders（6.24±2.54）（P<0.05）. Fifty-three patients were followed up for 0.5 to 1.8 years after operation, and in 43 cases the tinnitus was reduced or disappeared after operation（the effective rate was 81.13%）. There were no significant difference between patients in tinnitus relief group and those in tinnitus without relief group in age, sex, course of the disease, type of the disease, with or without comorbid psychiatric disorders and/or sleep disorder, postoperative hearing improvement. Conclusion:Comorbid psychiatric disorders are common in patients with chronic otitis media associated tinnitus and the tinnitus in patients with comorbid psychiatric disorders is significantly more serious than that those without. For the treatment of chronic otitis media associated tinnitus, besides surgery, the complications such as psychiatric and sleep disorders and so on should be evaluated and treated accordingly.

[The effect of one-stage tympanoplasty for stapes fixation with tympanosclerosis].

Objective:To evaluate hearing outcome and complications of one-stage tympanoplasty in patients with stapes fixation due to tympanosclerosis. Method:59 patients（sixty-one ears） underwent one-stage tympanoplasty for stapes fixation due to tympanosclerosis were retrospectively analyzed. Stapes fixation due to tympanosclerosis were proved during the surgery in these patients diagnosed with chronic otitis media. For all the patients, tympanosclerotic plaques around stapes were removed for stapes mobilization. Then the ossicular chain was rebuilt by autogenous incus or PORP. The pre-and post-operative audiometric results（500 Hz, 1 kHz, 2 kHz and 4 kHz） were evaluated for each patient. Improvement of pure-tone average more than 10 dB postoperatively were accepted as success criteria. Result:Complications included temporary facial paralysis（1/61）, temporary vertigo（2/61）, mild elevation in bone conduction thresholds（2/61） and delayed healing of tympanic membrane（1/61）. Postoperative（1 and 3 months） bone conduction thresholds improved at frequencies of 1 and 2 kHz（P<0.01）. Postoperative（1 and 2 years） air conduction thresholds improved at all frequencies（P<0.01 or P<0.05）. A gain ≥10 dB in pure-tone average was found in 44（72.13%） patients at 1 year after surgery. The air conduction levels were significantly improved in both autogenous incus and PORP groups（P<0.01）. There was no difference about success rate between these two groups（P>0.05）. Conclusion:For patients with stapes fixation due to tympanosclerosis, one-stage tympanoplasty can improve hearing threshold though ossicular chain reconstruction and stapes release. The major complications such as facial paralysis and sensorineural hearing loss should be avoided by delicate surgical operation.

The Msi1-mTOR pathway drives the pathogenesis of mammary and extramammary Paget's disease.

Mammary and extramammary Paget's Diseases (PD) are a malignant skin cancer characterized by the appearance of Paget cells. Although easily diagnosed, its pathogenesis remains unknown. Here, single-cell RNA-sequencing identified distinct cellular states, novel biomarkers, and signaling pathways - including mTOR, associated with extramammary PD. Interestingly, we identified MSI1 ectopic overexpression in basal epithelial cells of human PD skin, and show that Msi1 overexpression in the epidermal basal layer of mice phenocopies human PD at histopathological, single-cell and molecular levels. Using this mouse model, we identified novel biomarkers of Paget-like cells that translated to human Paget cells. Furthermore, single-cell trajectory, RNA velocity and lineage-tracing analyses revealed a putative keratinocyte-to-Paget-like cell conversion, supporting the in situ transformation theory of disease pathogenesis. Mechanistically, the Msi1-mTOR pathway drives keratinocyte-Paget-like cell conversion, and suppression of mTOR signaling with Rapamycin significantly rescued the Paget-like phenotype in Msi1-overexpressing transgenic mice. Topical Rapamycin treatment improved extramammary PD-associated symptoms in humans, suggesting mTOR inhibition as a novel therapeutic treatment in PD.

Secreted stromal protein ISLR promotes intestinal regeneration by suppressing epithelial Hippo signaling.

The Hippo-YAP signaling pathway plays an essential role in epithelial cells during intestinal regeneration and tumorigenesis. However, the molecular mechanism linking stromal signals to YAP-mediated intestinal regeneration and tumorigenesis is poorly defined. Here, we report a stroma-epithelium ISLR-YAP signaling axis essential for stromal cells to modulate epithelial cell growth during intestinal regeneration and tumorigenesis. Specifically, upon inflammation and in cancer, an oncogenic transcription factor ETS1 in stromal cells induces expression of a secreted protein ISLR that can inhibit Hippo signaling and activate YAP in epithelial cells. Deletion of Islr in stromal cells in mice markedly impaired intestinal regeneration and suppressed tumorigenesis in the colon. Moreover, the expression of stromal cell-specific ISLR and ETS1 significantly increased in inflamed mucosa of human IBD patients and in human colorectal adenocarcinoma, accounting for the epithelial YAP hyperactivation. Collectively, our findings provide new insights into the signaling crosstalk between stroma and epithelium during tissue regeneration and tumorigenesis.

Mucoadhesive-to-penetrating controllable peptosomes-in-microspheres co-loaded with anti-miR-31 oligonucleotide and Curcumin for targeted colorectal cancer therapy.

Background: Accumulating evidences indicate that nanomedicines greatly decrease the side effects and enhance the efficacy of colorectal cancer (CRC) treatment. In particular, the use of rectal delivery of nanomedicines, with advantages such as fast therapeutic effects and a diminishing hepatic first-pass effect, is currently emerging. Method: We established a CRC targeted delivery system, in which α-lactalbumin peptosomes (PSs) co-loaded with a microRNA (miR)-31 inhibitor (miR-31i) and curcumin (Cur) were encapsuslated in thiolated TEMPO oxidized Konjac glucomannan (sOKGM) microspheres, referred as sOKGM-PS-miR-31i/Cur. The CRC targeting capability, drug release profiles, mucoadhesive-to-penetrating properties and therapeutic efficacy of sOKGM-PS-miR-31i/Cur delivery system were evaluated in colorectal cancer cells and azoxymethane-dextran sodium (AOM-DSS) induced tumor models. Results: sOKGM-PS-miR-31i/Cur delivery system were stable in the harsh gastrointestinal environment after rectal or oral administration; and were also mucoadhesive due to disulfide bond interactions with the colonic mucus layer, resulting in an enhanced drug retention and local bioavailability in the colon. Concomitantly, the released PS-miR-31i/Cur PSs from the microsphere was mucus-penetrating, efficiently passing through the colonic mucus layer, and allowed Cur and miR-31i specifically target to colon tumor cells with the guide of CD133 targeting peptides. Consequently, rectal delivery of sOKGM-PS-miR-31i/Cur microspheres suppressed tumor growth in an azoxymethane-dextran sodium sulfate (AOM-DSS)-induced tumor model. Conclusion: sOKGM-PS-miR-31i/Cur microspheres are effective rectal delivery system with combined advantages of mucoadhesive and mucus-penetrating properties, representing a potent and viable therapeutic approach for CRC.

Fufang Xueshuantong protects retinal vascular endothelial cells from high glucose by targeting YAP.

Fufang Xueshuantong (FXST), a Chinese patent medicine, is composed of Panax notoginseng, Salviae miltiorrhizae, Astragali Radix and Radix Scrophulariae and has been found to prevent diabetic retinopathy. Yes-associated protein (YAP) participates in the pathophysiology of retinal disease and promotes endothelial cell proliferation and angiogenesis. Although it is known that YAP activity is altered by FXST, the role of YAP in mediating the effect of FXST remains unclear. In high glucose-treated retinal vascular endothelial cells (RVECs), FXST significantly reduced cell viability, the number of migrating cells and tube length in the present study. Moreover, FXST decreased the levels of YAP mRNA and protein and inhibited the expression of vascular endothelial growth factor (VEGF). Transfection of sh-YAP into the cells decreased the ability of FXST to modulate cell migration and tube formation. The effect of FXST on VEGF expression was also decreased. Similar results were obtained when the cells were stimulated with a YAP inhibitor in combination with FXST. Thus, FXST is shown to protect high glucose-injured RVECs via YAP-mediated effects.

[Overexpression of CGRP improves peripheral neuropathy in diabetic mice].

Objective To observe the effect of calcitonin gene-related peptide (CGRP) on peripheral neuropathy in diabetic mice, and to explore whether overexpressed CGRP improves peripheral neuropathy in diabetic mice. Methods Diabetic mouse model was induced by intraperitoneal injection of streptozotocin (STZ). The mice were randomly divided into 4 groups according to their body mass and blood glucose: normal control group, model group, Ad-CGRP group and Ad negative control group. The mice in the Ad-CGRP group and the Ad-negative control group were intramuscularly injected with adenovirus in the muscles of left hind limb. The mice in the normal group and the model group were injected with equal volume of normal saline. After 4 weeks of administration, the expression of CGRP mRNA was detected by real-time quantitative PCR. The expression of CGRP protein was detected by Western blot analysis. The latency of pain was measured by hot plate test. The motor nerve conduction velocity (MNCV) was measured by physiological signal acquiring system. The nerve blood flow was monitored with Laser Doppler Flowmetry (LDF). Results Compared with the normal group, the mRNA and protein levels of CGRP in the model group were reduced, the level of glucose was significantly raised, the latency of pain was extended, the MNCV and the nerve blood flow were decreased. Compared with the model group, the mRNA and protein levels of CGRP in the Ad-CGRP group were significantly increased, the latency of pain was shortened, the MNCV and the nerve blood flow was increased. There was no significant change in the Ad-negative control group. Conclusion Overexpression of CGRP may ameliorate diabetic peripheral neuropathy by expanding blood vessel and increasing blood flow.