Obesity is a prevalent metabolic disorder, which seriously affects human health and has become the world's public health problem. Kinase S6K1, an important downstream effector of mammalian target of rapamycin (mTOR), influences specific pathological responses, including obesity, type 2 diabetes and cancer. Presently, S6K1 has become an attractive therapeutic target in the treatment of these disorders. Here, the functions of kinase S6K1, its molecular regulation mechanisms, related pathogenesis of disease and relevant small molecular inhibitors are reviewed. Finally, the prospect of research toward S6K1 is expected as well.
Animals ; Diabetes Mellitus, Type 2 ; Humans ; Neoplasms ; Obesity ; Ribosomal Protein S6 Kinases, 70-kDa ; metabolism ; Signal Transduction ; TOR Serine-Threonine Kinases ; metabolism

Objective To study the anti-inflammatory effects of idazoxan (IDA) on endotoxin lipopolysaccharide (LPS) challenged mice in vivo and activated macrophages in vitro,and explore its potential molecular mechanisms.Methods To do the experiments in vivo,30 adult male C57BL/6 mice were randomly divided into control group,model group,and low,medium and high doses IDA groups (IDA-L,IDA-M,and IDA-H groups),n =6 in each group.The inflammatory model was reproduced by intraperitoneal injection of LPS 10 mg/kg,and the control group was injected with the same amount of normal saline.The IDA groups received LPS (10 mg/kg) and IDA 0.3,1.0 and 3.0 mg/kg,respectively.The blood samples of mice in each group were collected at 6 hours after the reproduction of the model.For the in vitro experiments,primary peritoneal macrophages were collected from 20 adult male C57BL/6 mouse cells and they were divided into control group,LPS group (10 mg/L) and LPS+IDA-L,IDA-M,IDA-H groups (10 mg/L LPS + 5,25,100 μmol/L IDA,respectively).Cell culture supernatants were collected at 24 hours after the reproduction of the model.Detection methods:enzyme linked immunosorbent assay (ELISA) was used to determine the levels of serum tumor necrosis factor-α (TNF-α),interleukin-6 (IL-6),monocyte chemotactic protein-1 (MCP-1) and nitric oxide (NO).Western Blot was used to determine the effect of IDA on the expression levels of nuclear factor-κB (NF-κB) in macrophages.Results ① For the in vivo experiment,the serum levels of TNF-α and IL-6 were significantly elevated in the model group as compared with those in the control group [TNF-o (ng/L):403.96 ± 40.98 vs.17.50 ± 8.68;IL-6 (ng/L):61 400.31 ± 7 826.61 vs.2 436.30 ± 448.89;both P ＜ 0.01].IDA treatment could inhibit the elevation of inflammatory cytokines in a dose-dependent manner,with the most significant decrease in LPS+IDA-H group [TNF-α (ng/L):170.09 ± 28.53 vs.403.96 ± 40.98,IL-6 (ng/L):16 570.81 ± 1 083.65 vs.61 400.31± 7 826.61;both P ＜ 0.01].② For the in vitro experiment,the levels of TNF-α,IL-6,MCP-1,and NO secreted by LPS-stimulated macrophages were distinctly higher in the LPS group than those in the control group [TNF-α (ng/L):7 259.14 ± 320.70 vs.28.50±27.08,IL-6 (ng/L):14809.60±5852.73 vs.1 113.47±465.53,MCP-1 (ng/L):20847.37± 1 788.33 vs.447.37± 395.69,NO (μmol/L):1 900.00 ± 144.31 vs.603.03 ± 102.18;all P ＜ 0.01].However,IDA intervention could lower the secretion of TNF-α,IL-6,MCP-1 and NO in a dose-dependent manner,with the most notable decrease in the LPS+IDA-H group [TNF-α (ng/L):784.40±281.90 vs.7259.14±320.70,IL-6 (ng/L):1 802.96± 1 534.18 vs.14 809.60± 5 852.73,MCP-1 (ng/L):2005.26± 1 534.28 vs.20847.37 ± 1 788.33,NO (μ mol/L):654.54± 150.21 vs.1 900.00 ± 144.31;all P ＜ 0.05].In addition,IDA at the concentration of 100 μmol/L could promote the translocation of NF-κBp65 in macrophages into the nucleus 15 minutes early and lead to increased NF-κBp65 expression (gray value:18.70 ± 2.29 vs.1.09 ± 0.36,P ＜ 0.05),hut significantly reduce the expression levels of NF-κBp50 in the nucleus at 45 minutes after treatment (gray value:1.99 ± 0.14 vs.2.94 ± 0.54,P ＜ 0.05).Conclusions IDA could significantly reduce inflammation of mice challenged with LPS and inhibit inflammatory cytokines and mediators secreted by macrophage in a dose-dependent manner.High concentration of IDA (100 μmol/L) exhibited the greatest anti-inflammatory effects.The anti-inflammatory effect of IDA may be worked through NF-κB signaling pathway.

Objective:To evaluate the effect of surgery on 47 patients with moyamoya disease by retrospective analysis.Methods:A total of 47 patients with moyamoya disease were enrolled from August,2010 to According to the improved treatment in August,2013,all cases were divided into two groups:a pre-improved group and a post-improved group.According to different surgical methods,they were divided into two subgroups:an indirect revascularization subgroup and a combined revascularization subgroup.Results:The cerebral ischemia in 77.4％ of patients was relieved after the surgery.There was significant difference in outcomes of patients between the pre-improved group and the post-improved group (P＜0.05),while there was no significant difference between the pre-improved indirect revascularization subgroup and the pre-improved combined revascularization subgroup.There was also no significant difference between the post-improved indirect revascularization subgroup and the post-improved combined revascularization subgroups (P＞0.05).Conclusion:Surgical treatment can improve the outcomes of patients with moyamoya disease,but there is no significant difference in surgical effects between indirect and combined revascularization.

Objective To verify that curcumol can inhibit erastin-induced ferroptosis in HT22 cells and play a role of cell protection.Methods Firstly,the effect of 0.5 μmol·L-1 Erastin at different time gradients(0,4,8,12,24 h)on the expression of ferroptosis-related proteins in HT22 cells were abserved by Western blot,Then HT22 cells were divided into normal control group,DMSO group,Erastin treatment group(0.5 μmol·L-1 Erastin treatment for 24),and curcumol group(0.5 μmol·L-1 Erastin treatment for 24 h+50 μmol·L-1 curcumin treatment for 48 ho).The expression levels of ferroptosis-related proteins in HT22 cells were detected by Western blot,cell iron ion kit and cell ferrous content kit were used to detect the levels of total iron and ferrous iron in cells,cell viability was detected by CCK-8 kit,and cell apoptosis was detected by Annexin V/PI apoptosis kit.Results After Erastin stimulation,the levels of xCT(P<0.05),NCOA4(P<0.001)and ACSL4(P<0.01)were up-regulated,while GPX4(P<0.001),FPN(P<0.01)and FTH1(P<0.05)were decreased in HT22 cells.The ferritin-related protein levels,iron levels,cell viability and apoptosis rate of DMSO group were basically the same as those of normal control group.Compared with Erastin group,the cell viability of neurons(P<0.001)and the GPX4 enzyme level of the cells(P<0.01)in the curcumol treatment group was significantly increased,the apoptosis rate(P<0.001)and the total iron and ferrous iron contents of the cells(P<0.001)were decreased oppositely.In addition,the expression levels of Xct(P<0.001),NCOA4(P<0.001),and ACSL4(P<0.001)were significantly down-regulated,the expression levels of FTH1(P<0.01)and GPX4(P<0.001)were significantly up-regulated(P<0.01).Conclusion Curcumol could protect HT22 cells by inhibiting erastin-induced ferroptosis.

Objective:To explore the effects of neutrophilic granule protein (NGP) on the expression of lipocalin 2 (LCN2) in inflammatory macrophages and its mechanism.Methods:NGP-high-expressed RAW264.7 cells (NGP/RAW cells) and negative control RAW264.7 cells (NC/RAW cells) were cultured in vitro. Primary peritoneal macrophages of NGP-high-expressed mice and wild-type C57BL/6 mice were extracted, then cultured in vitro. The cell inflammatory model was established by stimulating with 10 mg/L lipopolysaccharide (LPS, LPS group), and the phosphate buffer solution (PBS) control group was set up. Enzyme-linked immunosorbent assay (ELISA) was used to detect the level of LCN2 in different types of cells. The protein expression of phosphorylated signal transduction and activator of transcription 1 (p-STAT1) was detected with Western blotting. Other NGP/RAW cells and NC/RAW cells were treated with 10 mg/L LPS, 5 mg/L STAT1 pathway inhibitor (fludarabine)+10 mg/L LPS, respectively. The PBS control group was set up. ELISA was used to detect the level of LCN2. Results:In different types of cells, the levels of LCN2 were increased significantly after LPS stimulation in the LPS group as compared with those in the PBS control group, and peaked at 24 hours (μmol/L: 25.61±1.02 vs. 0.46±0.02 in NC/RAW cells, 74.51±2.14 vs. 0.25±0.04 in NGP/RAW cells, 10.13±0.22 vs. 0.01±0.01 in primary macrophages of wild-type C57BL/6 mice, 28.35±0.61 vs. 0.08±0.01 in primary macrophages of NGP-high-expressed mice, all P < 0.05), indicating that the expression of LCN2 in macrophages altered during inflammation reaction. The level of LCN2 in NGP/RAW cells was found significantly increased at different time points after LPS stimulation comparing with that in NC/RAW cells (μmol/L: 8.32±0.22 vs. 3.12±0.11 at 6 hours, 23.12±0.86 vs. 8.12±0.32 at 12 hours, 74.51±2.14 vs. 25.61±1.02 at 24 hours, all P < 0.05), along with the expression of p-STAT1 was significantly up-regulated. The level of LCN2 in the primary macrophages of NGP-high-expressed mice was also significantly increased at 24 hours after LPS stimulation comparing with that in the primary macrophages of wild-type C57BL/6 mice (μmol/L: 28.35±0.61 vs. 10.13±0.22, P < 0.05). However, after pretreated with STAT1 pathway inhibitors, the production of LCN2 in NGP/RAW cells was decreased significantly comparing with that in the LPS group (μmol/L: 6.81±0.19 vs. 22.54±0.58, P < 0.05). But the inhibitors had no significant effect on LCN2 production in NC/RAW cells showing no significant difference as compared with LPS group (μmol/L: 8.04±0.20 vs. 7.86±0.15, P > 0.05), indicating that NGP could up-regulate the expression of LCN2 in macrophages stimulated by LPS by promoting STAT1 activation. Conclusion:NGP could positively regulate LCN2 expression in inflammatory macrophages by activating STAT1 pathway.

Objective:To explore the clinical features and risk factors of in-hospital mortality in idiopathic inflammatory myopathies (IIM) patients.Results:We retrospectively analyzed clinical records of polymyositis (PM), classic dermatomyositis (CDM) and clinically amyopathic dermatomyositis (CADM) patients admitted to the First Affiliated Hospital of Zhejiang University from February 2011 to February 2019. The deceased group was defined as the patients who died in hospital or within 2 weeks after hospital discharge, while the survival group was defined as the survival patients. The clinical features were described. Risk factors for deceased patients were identified by logistic regression analysis.Results:The in-hospital mortality rate of IIM patients ( n=424) was 9.4%. The hospitalization time was longer in deceased group ( n=40) [0.9(0.5, 1.0) m vs 0.6(0.4, 1.0) m, Z=-2.159, P<0.05]. Ferritin [1170.8(757.6, 3 759.9) μg/L vs 374.9(182.1, 993.4) μg/L, Z=-4.665], red blood cell distribution width (RDW) [15.2(14.5, 16.3)% vs 14.4(13.5, 15.2)%, Z=-3.066], CRP con-centrations [11.3(4.4, 36.9) mg/L vs 5.1(1.8, 17.2) mg/L, Z=-2.667] and neutrophil-to-lymphocyte ratio (NLR) [10.1(5.5, 18.9) vs 4.2(2.6, 6.5), Z=-5.108] were higher in deceased group ( P<0.05). Proportion of patients with high levels of CEA (45.0% vs 12.5%, χ2=15.745), glutamyl transpeptidase (γ-GT) (55.0% vs 23.8%, χ2=11.578), fucosidase (AFU) (35.0% vs 10.0%, χ2=10.902) and with complications [including pro-gressive in-terstitial lung disease (ILD) (60.0% vs 16.3%, χ2=23.934), pulmonary infection (72.5% vs 20.0%, χ2=31.360), hemophagocytic lymphohistiocytosis (35.0% vs 1.3%, χ2=27.771) and low T3 syndrome (50.0% vs 17.5%, χ2=16.644) were higher in deceased group ( P<0.05). Steroid pulse therapy and intravenous immuno-globulin therapy were more common in deceased group. Higher on-admission disease activity [ OR=1.593, 95% CI(1.255, 2.022), P<0.001], progressive ILD [ OR=5.600, 95% CI(1.510, 20.772), P=0.010] and pulmonary infection [ OR=6.771, 95% CI(2.031, 22.574), P=0.002] were independent risk factors for death in IIM patients. In su-bsection analysis, pulmonary infection and respiratory failure were short-term adverse prognostic factors for IIM patients with progressive ILD, while heliotrope rash, progressive ILD and increased steroid dose therapy were short-term adverse prognostic factors for IIM patients with pulmonary infection. Conclusion:High disease activity at admission, progressive ILD and pulmonary infection are the independent risk factors for death in IIM patients. Therefore, it is necessary to closely monitor above indicators during hospitalization.

Fragile X syndrome (FXS) is a neurodevelopmental synaptopathy caused by loss of fragile X mental retardation protein (FMRP); abnormal synaptic plasticity is the leading pathological cause of cognitive impairment, fear and anxiety, hyperactivity and stereotyped behavior in FXS patients. In recent years, breakthroughs have been made in functional study of synaptic plasticity in FXS, providing a new theoretical basis for FXS. This article mainly summarizes the dysregulation and influencing factors of synaptic plasticity in FXS, as well as the strategy of targeted synaptic plasticity in FXS, so as to deepen the understanding of medical workers.

Objective:To explore the expression pattern of aryl hydrocarbon receptor (AhR) in mice peritoneal macrophages (PMs) after major trauma and analyze the effects of enhanced AhR expression on the inflammatory cytokine level and bactericidal ability after trauma.Methods:The experimental study method was used. Forty 6-8-week-old male C57BL/6J mice (the same mouse age, sex, and strain below) were divided into control group, post trauma hour (PTH) 2 group, PTH 6 group, and PTH 12 group according to the random number table (the same grouping method below), with 10 mice in each group. Mice in the latter 3 groups were constructed as severe trauma model with fracture+blood loss, while mice in control group were left untreated. The primary PMs (the same cells below) were extracted from the mice in control group, PTH 2 group, PTH 6 group, and PTH 12 group when uninjured or at PTH 2, 6, and 12, respectively. Then the protein and mRNA expressions of AhR were detected by Western blotting and real-time fluorescence quantitative reverse transcription polymerase chain reaction, respectively, and the gene expressions of AhR signaling pathway related molecules were analyzed by transcriptome sequencing. Twenty mice were divided into control group and PTH 6 group, with 10 mice in each group, and the PMs were extracted. The level of ubiquitin of AhR was detected by immunoprecipitation. Twelve mice were divided into dimethyl sulfoxide (DMSO) alone group, PTH 6+DMSO group, MG-132 alone group, and PTH 6+MG-132 group, with 3 mice in each group. After the corresponding treatment, PMs were extracted, and the protein expression of AhR was detected by Western blotting. Twenty mice were constructed as PTH 6 model. Then, the PMs were extracted and divided into empty negative control adenovirus (Ad-NC) group and AhR overexpression adenovirus (Ad-AhR) group. The protein expression of AhR was detected by Western blotting at 36 h after some PMs were transfected with the corresponding adenovirus. The rest cells in Ad-NC group were divided into Ad-NC alone group and Ad-NC+endotoxin/lipopolysaccharide (LPS) group, and the rest cells in Ad-AhR group were divided into Ad-AhR alone group and Ad-AhR+LPS group. The expressions of interleukin-6 (IL-6) and tumor necrosis factor α (TNF-α) in the cell supernatant were detected by enzyme-linked immunosorbent assay at 12 h after the corresponding treatment ( n=6). Twenty mice were obtained to extract PMs. The cells were divided into control+Ad-NC group, PTH 6+Ad-NC group, control+Ad-AhR group, and PTH 6+Ad-AhR group, and the intracellular bacterial load was detected by plate spread method after the corresponding treatment ( n=6). Data were statistically analyzed with one-way analysis of variance, least significant difference test, analysis of variance for factorial design, and independent sample t test. Results:Compared with 1.16±0.28 of control group, the protein expressions of AhR in PMs in PTH 2 group (0.59±0.14), PTH 6 group (0.72±0.16), and PTH 12 group (0.71±0.17) were all significantly decreased ( P<0.05). The overall comparison of the difference of AhR mRNA expression in PMs among control group, PTH 2 group, PTH 6 group, and PTH 12 group showed no statistical significance ( P>0.05). The AhR signaling pathway related molecules included AhR, AhR inhibitor, cytochrome P450 family member 1b1, cytochrome P450 family member 11a1, heat shock protein 90, aryl hydrocarbon receptor-interaction protein, and heat shock protein 70 interaction protein. The heat shock protein 90 expression of PMs in PTH 2 group was higher than that in control group, while the expressions of other molecules did not change significantly after trauma. Compared with that in control group, the level of ubiquitin of AhR in PMs in PTH 6 group was increased. Compared with that in DMSO alone group, the protein expression of AhR in PMs in PTH 6+DMSO group was decreased, while that in PMs in MG-132 alone group had no significant change. Compared with that in PTH 6+DMSO group, the protein expression of AhR in PMs in PTH 6+MG-132 group was up-regulated. At transfection hour 36, compared with that in Ad-NC group, the protein expression of AhR in PMs in Ad-AhR group was increased. At treatment hour 12, compared with those in Ad-NC+LPS group, the expressions of IL-6 and TNF-α in PM supernatant of Ad-AhR+LPS group were significantly decreased (with t values of 4.80 and 3.82, respectively, P<0.05). The number of intracellular bacteria of 1×10 6 PMs in control+Ad-NC group, PTH 6+Ad-NC group, control+Ad-AhR group, and PTH 6+Ad-AhR group was (3.0±1.8), (41.8±10.2), (1.8±1.2), and (24.2±6.3) colony forming unit, respectively. Compared with that in PTH 6+Ad-NC group, the number of intracellular bacteria of PMs in PTH 6+Ad-AhR group was significantly decreased ( t=3.61, P<0.05). Conclusions:Ubiquitin degradation of AhR in PMs of mice after major trauma results in decreased protein expression of AhR. Increasing the expression of AhR in post-traumatic macrophages can reduce the expressions of LPS-induced inflammatory cytokines IL-6 and TNF-α, and improve the bactericidal ability of macrophages after trauma.

In this study, we attempted to establish a culture system for in vitro spermatogenesis from spermatogonial stem cells (SSCs) of Bama mini-pig. Dissociated testicular cells from 1-month-old pigs were co-cultured to mimic in vivo spermatogenesis. The testicular cells were seeded in minimum essential medium alpha (α-MEM) supplemented with Knockout serum replacement (KSR). Three-dimensional colonies formed after 10 days of culture. The colonies showed positive staining for SSC-associated markers such as UCHL1, PLZF, THY1, OCT4, Dolichos biflorus agglutinin, and alkaline phosphatase. Induction of SSCs was performed in α-MEM + KSR supplemented with retinoic acid, bone morphogenetic protein 4, activin A, follicle-stimulating hormone, or testosterone. The results showed that STRA8, DMC1, PRM1, and TNP1 were upregulated significantly in the colonies after induction compared to that in testis from 1-month-old pigs, while expression levels of those genes were significantly low compared to those in 2-month-old testis. However, upregulation of ACROSIN was not significant. Replacement of α-MEM and KSR with Iscove's modified Dulbecco's medium and fetal bovine serum did not upregulate expression of these genes significantly. These results indicate that SSCs of Bama mini-pig could undergo differentiation and develop to a post-meiotic stage in α-MEM supplemented with KSR and induction factors.
Acrosin ; Activins ; Alkaline Phosphatase ; Bone Morphogenetic Protein 4 ; Dolichos ; Follicle Stimulating Hormone ; Humans ; In Vitro Techniques ; Infant ; Infant, Newborn ; Spermatogenesis ; Stem Cells ; Swine ; Testis ; Testosterone ; Tretinoin ; Up-Regulation

Sepsis remains the leading cause of late death in trauma patients. Many bottleneck problems in the field of post-traumatic sepsis research have not been solved, which are mainly reflected in the lag of early warning research, limited preventive interventions and unclear diagnostic criteria. The authors focus on the early warning of sepsis, preventive intervention measures and early diagnostic criteria to guide the clinical positive response so as to reduce the late mortality caused by post-traumatic sepsis and improve the overall level of trauma treatment.