Liver-specific Reduction of Mfn2 Protein by RNAi Results in Impaired Glycometabolism and Lipid Homeostasis in BALB/c Mice

Mitofusin-2 （Mfn2） gene expression is positively correlated with insulin sensitivity in patients with type 2 diabetes. However, it is unclear if Mfn2 is involved in carbohydrate metabolism and lipid homeostasis. In order to investigate the specific functions of Mfn2 in glycometabolism and lipid homeostasis in BALB/c mice, a RNA interference technique-mediated hydrodynamic injection was developed, in which short hairpin RNAs （shRNAs） were used to inhibit the Mfn2 expression in vivo. Seventy-two mice were randomly divided into two groups： the Mfn2 reduction group （Mfn2/shRNA） and the negative control group （NC）. Intraperitoneal glucose tolerance tests and intraperitoneal insulin tolerance tests were used to evaluate glycometabolism and insulin sensitivity. D-（3-3H） glucose or 3H2O was injected into the tail vein or intraperitoneally to facilitate the calculation of the rate of hepatic glucose production and fatty acid synthesis in vivo. The results showed that, in Mfn2/shRNA mice, the liver Mfn2 protein was significantly decreased, and fasting blood glucose concentrations were increased by approximately 48%, when compared with the NC mice. In parallel with the changes in fasting glucose levels, hepatic glucose production was significantly elevated in Mfn2/shRNA mice. When insulin was administrated, these mice exhibited impaired insulin tolerance. It was also found that the reduction of Mfn2 markedly decreased the rate of fatty acid synthesis in the liver, and the Mfn2/shRNA mice exhibited hypertriglyceridema. Taken together, our results indicate that Mfn2 plays an important role in maintaining glucose and lipid homeostasis, and in the development of insulin resistance in vivo.

Basal physiological parameters in domesticated tree shrews(Tupaia belangeri chinensis)

Establishing non-human primate models of human diseases is an efficient way to narrow the large gap between basic studies and translational medicine.Multifold advantages such as simplicity of breeding,low cost of feeding and facility of operating make the tree shrew an ideal non-human primate model proxy.Additional features like vulnerability to stress and spontaneous diabetic characteristics also indicate that the tree shrew could be a potential new animal model of human diseases.However,basal physiological indexes of tree shrew,especially those related to human disease,have not been systematically reported.Accordingly,we established important basal physiological indexes of domesticated tree shrews including several factors:(1)body weight,(2)core body temperature and rhythm,(3)diet metabolism,(4)locomotor rhythm,(5)electroencephalogram,(6)glycometabolism and(7)serum and urinary hormone level and urinary cortisol rhythm.We compared the physiological parameters of domesticated tree shrew with that of rats and macaques.Results showed that(a)the core body temperature of the tree shrew was 39.59±0.05°C,which was higher than that of rats and macaques;(b)Compared with wild tree shrews,with two activity peaks,domesticated tree shrews had only one activity peak from 17:30 to 19:30;(c)Compared with rats,tree shrews had poor carbohydrate metabolism ability;and(d)Urinary cortisol rhythm indicated there were two peaks at 8:00 and 17:00 in domesticated tree shrews,which matched activity peaks in wild tree shrews.These results provided basal physiological indexes for domesticated tree shrews and laid an important foundation for diabetes and stress-related disease models established on tree shrews.

Detection of serum Betatrophin in gestational diabetes mellitus patients and its correlation with glucose and lipid metabolism disorders

Objective: To detect the serum Betatrophin level in patients with gestational diabetes mellitus (GDM), and to explore the relationship between Betatrophin level and disorders of glucose and lipid metabolism. Methods: 119 cases of GDM women in our hospital during August 2015 to March 2018 were selected as GDM group, 100 cases pregnant women with normal blood glucose were chosen as Normal control group. Serum level of Betatrophin, glucose metabolism and lipid metabolism between two groups were compared. Pearson test was used to evaluate the relationship between Betatrophin, glucose and lipid metabolism disorders in GDM pregnant women. Results: Serum content of Betatrophin in GDM group was significantly higher than that in Normal control group, levels of HbA1c, FINS and HOMA-IR in peripheral blood were significantly higher than those in Normal control group;contents of TC, LDL-C and Apo-B in peripheral blood were higher than those in Normal control group, contents of HDL-C, Apo-A1 were lower than those in Normal control group. Correlation analysis showed that serum level of Betatrophin in GDM pregnant women was positively correlated with the disorder of glucose and lipid metabolism. Conclusion: Abnormally high expression of Betatrophin in GDM pregnant women may be an important factor in promoting disorder of glucose and lipid metabolism.

Effect of continuous blood purification on inflammatory state, immune response and erythrocyte glycometabolism in patients with multiple injury and sepsis

Objective:To analysis the effect of continuous blood purification on inflammatory state, immune response and erythrocyte glycometabolism in patients with multiple injury and sepsis. Methods: A total of 78 patients with multiple injury and sepsis were randomly divided into observation group (n=39) and control group (n=39), control group received routine therapy, observation group received continuous blood purification treatment, and then the differences in inflammatory state, immune response, erythrocyte glycometabolism and other indexes were compared between the two groups after treatment.Results: Inflammatory factor hs-CRP, TNF-α, PCT, sTREM-1 and HBP content in serum of observation group after treatment were significantly lower than those of control group;Th1 cytokines IL-2 and IFN-γ content in serum were lower than those of control group while Th2 cytokines IL-4 and IL-10 content were higher than those of control group;PFK and EGSH content in erythrocyte solution were higher than those of control group while G-6PD, AR and ELPO content were lower than those of control group;fluorescence intensity of CD11a, CD54, CD106 and CD49d in peripheral blood were significantly lower than those of control group.Conclusions: Continuous blood purification can significantly reduce the systemic inflammatory response in patients with multiple injury and sepsis, and promote the immune function and erythrocyte metabolism to return to normal.

Click CAR-T cell engineering for robustly boosting cell immunotherapy in blood and subcutaneous xenograft tumor

The adoptive transfer of chimeric antigen receptor-T(CAR-T)cells has shown remarkable clinical responses in hematologic malignancies.However,unsatisfactory curative results and side effects for tumor treatment are still unsolved problems.Herein we develop a click CAR-T cell engineering strategy via cell glycometabolic labeling for robustly boosting their antitumor effects and safety in vivo.Briefly,paired chemical groups(N3/BCN)are separately incorporated into CAR-T cell and tumor via nondestructive intrinsic glycometabolism of exogenous Ac4GalNAz and Ac4ManNBCN,serving as an artificial ligand-receptor.Functional groups anchored on cell surface strengthen the interaction of CAR-T cell and tumor via bioorthogonal click chemistry,further enhancing specific recognition,migration and selective antitumor effects of CAR-T cells.In vivo,click CAR-T cell completely removes lymphoma cells and minimizes off-target toxicity via selective and efficient bioorthogonal targeting in blood cancer.Surprisingly,compared to unlabeled cells,artificial bioorthogonal targeting significantly promotes the accumulation,deep penetration and homing of CAR-T cells into tumor tissues,ultimately improving its curative effect for solid tumor.Click CAR-T cell engineering robustly boosts selective recognition and antitumor capabilities of CAR T cells in vitro and in vivo,thereby holding a great potential for effective clinical cell immunotherapy with avoiding adverse events in patients.

Validamycin treatment significantly inhibits the glycometabolism and chitin synthesis in the common cutworm,Spodoptera litura

Validamycin,as a broadly applied antibiotic,has been used to control rice sheath blight disease.Furthermore,validamycin was considered as an insecticide to control agricultural pests.Insight into the mechanism of validamycin’s action on insects can provide molecular targets for the control of agricultural pests.In this study,a toxicological test analysis revealed that Spodoptera litura larval growth and development was significantly inhibited and the pupation rate was significantly reduced with the increase of the concentration of validamycin.According to the NMR-based metabolomic analysis,a total of 15 metabolites involved in glycolysis and tricarboxylic acid cycle(TCA)pathways were identified.Additionally,trehalase activities,glucose and chitin contents were significantly downregulated,but the trehalose content was upregulated after exposure to validamycin.Reverse transcription quantitative PCR analysis revealed that the expression level of genes involved in glycolysis,TCA and chitin synthesis were upregulated after treating with validamycin.Further chitin staining also confirmed that chitin content was downregulated at 12 h after validamycin treatment.Our results indicated that validamycin worked via two different molecular mechanisms,one through inhibiting glycometabolism and the other by inhibiting chitin synthesis in S.litura.The information lays a theoretical foundation for further control of S.litura.