PPARγ-dependent hepatic macrophage switching acts as a central hub for hUCMSC-mediated alleviation of decompensated liver cirrhosis in rats.

BACKGROUND: Decompensated liver cirrhosis (DLC), a terminal-stage complication of liver disease, is a major cause of morbidity and mortality in patients with hepatopathies. Human umbilical cord mesenchymal stem cell (hUCMSC) therapy has emerged as a novel treatment alternative for the treatment of DLC. However, optimized therapy protocols and the associated mechanisms are not entirely understood. METHODS: We constructed a DLC rat model consistent with the typical clinical characteristics combined use of PB and CCL4. Performing dynamic detection of liver morphology and function in rats for 11 weeks, various disease characteristics of DLC and the therapeutic effect of hUCMSCs on DLC in experimental rats were thoroughly investigated, according to ascites examination, histopathological, and related blood biochemical analyses. Flow cytometry analysis of rat liver, immunofluorescence, and RT-qPCR was performed to examine the changes in the liver immune microenvironment after hucMSCs treatment. We performed RNA-seq analysis of liver and primary macrophages and hUCMSCs co-culture system in vitro to explore possible signaling pathways. PPARγ antagonist, GW9662, and clodronate liposomes were used to inhibit PPAR activation and pre-exhaustion of macrophages in DLC rats' livers, respectively. RESULTS: We found that changing the two key issues, the frequency and initial phase of hUCMSCs infusion, can affect the efficacy of hUCMSCs, and the optimal hUCMSCs treatment schedule is once every week for three weeks at the early stage of DLC progression, providing the best therapeutic effect in reducing mortality and ascites, and improving liver function in DLC rats. hUCMSCs treatment skewed the macrophage phenotype from M1-type to M2-type by activating the PPARγ signaling pathway in the liver, which was approved by primary macrophages and hUCMSCs co-culture system in vitro. Both inhibition of PPARγ activation with GW9662 and pre-exhaustion of macrophages in DLC rats' liver abolished the regulation of hUCMSCs on macrophage polarization, thus attenuating the beneficial effect of hUCMSCs treatment in DLC rats. CONCLUSIONS: These data demonstrated that the optimal hUCMSCs treatment effectively inhibits the ascites formation, prolongs survival and significantly improves liver structure and function in DLC rats through the activation of the PPARγ signaling pathway within liver macrophages. Our study compared the efficacy of different hUCMSCs infusion regimens for DLC, providing new insights on cell-based therapies for regenerative medicine.

A straight-forward seed production technology system for foxtail millet (Setaria italica).

For autogamous crops, a precondition for using heterosis is to produce sufficient pure male-sterile female parents that can be used to produce hybrid seeds. To date, cytoplasmic male sterility (CMS) and environment-sensitive genic male sterility (EGMS) have been used commercially to exploit heterosis for autogamous species. However, neither CMS nor EGMS has been established for foxtail millet (Setaria italica). Here, we report on the establishment and application of a seed production technology (SPT) system for this crop. First, we established a DsRed-based SPT system, but found that it was unsuitable because it required the use of a fluorescent device for seed sorting. Instead, we constructed an SPT system with de novo betalain biosynthesis as the selection marker. This allowed us to distinguish transgenic seeds with the naked eye, thereby facilitating the identification of SPT maintainer line seeds. In this system, a seed sorter was not required to obtain sufficient seeds. The key point of the strategy is that the seed pool of the SPT maintainer line is propagated by artificial identification and harvesting of male-fertile individuals in the field, and the male-sterile line seed pool for hybrid production is produced and propagated by free pollination of male-sterile plants with the SPT maintainer line. In a field experiment, we obtained 423.96 kg male-sterile line seeds per acre, which is sufficient to plant 700.18 acres of farmland for hybrid seed production or male-sterile line reproduction. Our study therefore describes a powerful tool for hybrid seed production in foxtail millet, and demonstrates how the SPT system can be used for a small-grained crop with high reproduction efficiency.

Therapeutic efficacy of human adipose mesenchymal stem cells in Crohn's colon fibrosis is improved by IFN-γ and kynurenic acid priming through indoleamine 2,3-dioxygenase-1 signaling.

BACKGROUND: Inflammatory bowel diseases (IBD) are chronic relapsing-remitting inflammatory diseases of the gastrointestinal tract that are typically categorized into two subtypes: Crohn's disease (CD) and ulcerative colitis (UC). Although MSCs therapy has achieved encouraging outcomes in IBD therapy, objective responses are limited in colon fibrosis stenosis owing to the complicated microenvironment of CD and MSCs heterogeneity of quality. Here, we chose IFN-γ and kynurenic acid (KYNA) to overcome the low response and heterogeneity of human adipose-derived MSCs (hADSCs) to treat IBD and expand the therapeutic effects based on the excellent ability of IFN-γ and KYNA to promote indoleamine 2,3-dioxygenase-1 (IDO-1) signaling, providing a potential protocol to treat IBD and fibrosis disease. METHODS: hADSCs were isolated, cultured, and identified from human abdominal adipose tissue. The CD pathology-like acute colitis and chronic colon fibrosis rat model was induced by 2,4,6-trinitrobenzen sulfonic acid (TNBS). hADSCs were pretreated in vitro with IFN-γ and KYNA and then were transplanted intravenously at day 1 and 3 of TNBS administration in colitis along with at day 1, 15, and 29 of TNBS administration in chronic colonic fibrosis. Therapeutic efficacy was evaluated by body weights, disease activity index, pathological staining, real-time PCR, Western blot, and flow cytometry. For knockout of IDO-1, hADSCs were transfected with IDO-1-targeting small gRNA carried on a CRISPR-Cas9-lentivirus vector. RESULTS: hADSCs treated with IFN-γ and KYNA significantly upregulated the expression and secretion of IDO-1, which has effectively ameliorated CD pathology-like colitis injury and fibrosis. Notably, the ability of hADSCs with IDO-1 knockout to treat colitis was significantly impaired and diminished the protective effects of the primed hADSCs with IFN-γ and KYNA. CONCLUSION: Inflammatory cytokines IFN-γ- and KYNA-treated hADSCs more effectively alleviate TNBS-induced colitis and colonic fibrosis through an IDO-1-dependent manner. Primed hADSCs are a promising new strategy to improve the therapeutic efficacy of MSCs and worth further research.

3D Printing Mini-Capsule Device for Islet Delivery to Treat Type 1 Diabetes.

Transplantation of encapsulated islets has been shown to hold a promising potential treatment for type 1 diabetes (T1D). However, there are several obstacles to overcome, such as immune rejection by the host of the grafts, sustainability of islet function, and retrievability or replacement of the encapsulated system, hinder their clinical applications. In this study, mini-capsule devices containing islets were fabricated by using digital light processing (DLP) 3D printing. To ensure a high survival rate and low immunogenicity of the fabricated islets, 20s was selected as the most suitable printing condition. Meanwhile, the mini-capsule devices with a groove structure were fabricated to prevent islet cells leakage. Subcutaneous transplantations of encapsulated islets in immunocompetent C57BL/6 mice indicated significant improvement in the symptoms of streptozotocin-induced hyperglycemia without any immunosuppression treatment for at least 15 weeks. In vivo intraperitoneal glucose tolerance tests (IPGTT) performed at different time points demonstrated therapeutically relevant glycemic ameliorate of the device. The implants retrieved after 15 weeks still contained viable and adequate numbers of islet cells. The results of this study indicate that the proposed mini-capsule device can deliver sufficient islet cell mass, prevent islet cells leakage, and maintain long-term cell survival while allowing easy retrieval. Furthermore, the proposed encapsulated islets may help with T1D cellular treatment by overcoming the obstacles of islet transplantation.

Cp1-11 peptide/insulin complex loaded pH-responsive nanoparticles with enhanced oral bioactivity.

To improve the oral efficiency of insulin, a novel oral insulin delivery system was developed, to protect the insulin from destruction, and to deliver monomeric insulin with higher bioactivity. The oral insulin delivery system was developed by using chitosan/alginate nanoparticles as carriers for the oral delivery of Cp1-11 peptide/insulin complex (Cp1-11 peptide/Insulin Loaded Nanoparticle, CILN). There is a supramolecular interaction between the insulin and the Cp1-11 peptide, which could inhibit insulin aggregation and improve its bioactivity. In vitro release studies showed that the pH-responsive CILN system could retain insulin in a simulated gastric buffer solution, and exhibited sustained release of the insulin in simulated intestinal buffer solution. The stability studies indicated that CILN could also protect insulin against degradation by proteases in the gastrointestinal tract. Moreover, the insulin oral delivery system appeared to show excellent hypoglycemic effect, and led to higher pharmacological availability of insulin, compared with free insulin loaded nanoparticles, in diabetic rats. Thus, this study has suggested that insulin complexed with Cp1-11 peptide, followed by encapsulating in chitosan/alginate nanoparticles, providing an effective strategy for an oral insulin delivery system to treat diabetes in the future.

[Comparison of Relationship Between Conduction and Algal Bloom in Pengxi River and Modao River in Three Gorges Reservoir].

Pengxi River is the largest tributary in the northern bank of Yangtze River in the Three Gorges Reservoir (TGR) region, and serious algal bloom has been reported since 2003 when the TGR dam started impounding water. On the other hand, Modao River, a tributary in TGR locating in the same county with Pengxi River, was rarely reported with algal bloom in the same time period. In this study, water samples were collected on the same day in both rivers in spring and summer, the most likely blooming seasons in TGR, to compare the water quality parameters and Chlorophyll-a(Chl-a) concentration dynamics in order to understand the mechanism of algal bloom in TGR. The results showed that the maximum value of Chl-a in Pengxi river reached 60.5 µg·L-1 in spring, and was only 7.8 µg·L-1 in summer, while in Modao river the Chl-a content was 2.92 µg·L-1 in spring and 7.48 µg·L-1in summer. Both rivers stratified during samplings, but having layers of thermocline and hypolimnion with the lack of epilimnion, and most of Chl-a content was located in thermocline. Also in both rivers, TN and TP exceeded the international threshold for eutrophication, with the average TN and TP values of 2.305 mg·L-1 and 0.053 mg·L-1in Pengxi River in spring, and 1.673 mg·L-1 and 0.097 mg·L-1 in summer, respectively, while in Modao River the values were 1.875 mg·L-1 and 0.075 mg·L-1 in spring, and 1.79 mg·L-1 and 0.054 mg·L-1 in summer respectively. Nutrients concentrations showed no significant correlation with Chl-a. On the other hand, conductivity value and trend were totally different between the two rivers:in Modao River in spring, the conductivity in upstream was only 75% of that in the main stream of the Yangtze River, and the backwater from the main stream reached to the middle in Modao river, where the highest Chl-a among all the river sampling sites was detected; summer conductivity distribution was similar with that in spring. Different from Modao River, the conductivity in upstream of Pengxi River in spring was 150% of that in the main stream of Yangtze, the backwater from the main stream reached area between sampling sites of PX04 and PX05 (upper than the middle reach); its upstream had significantly high content of Chl-a and conductivity, and these two factors were significantly positively correlated. The study showed that other than N and P, other ions in the Pengxi River played an important role in bursting "bloom", and need to be considered regarding bloom control.