Effects of shift work on adipokines, inflammatory factors, and insulin sensitivity in non-overweight/obese women / 中华内分泌代谢杂志

Objective:To observe the effect of shift work on the stability of the circadian clock and insulin sensitivity in non-overweight/obese individuals with normal blood glucose, and explore underlying connection.Methods:Female shift working nurses in the Department of Blood Transplantation and non-shift working nurses in the Health Management Center in the First Affiliated Hospital of Zhengzhou University were divided into shift worker group (SW group) and non-shift worker group (NSW group). Serum inflammatory factors [interleukin-6 (IL-6), tumor necrosis factor-α(TNF-α)], adipokines (adiponectin, leptin, chemerin, visfatin), and melatonin levels were measured using enzyme linked immunosorbent assay (ELISA). Realtime fluorescence quantitative PCR was performed to detect peripheral blood circadian clock genes circadian locomotor output cycles protein kaput(Clock) and brain and muscle ARNT-like protein 1(Bmal1). Cortisol and fasting insulin were measured by chemiluminescent microparticle immunoassay, and HbA 1C was measured by capillary electrophoresis. In addition, visceral fat area (VFA) was assessed with bioelectrical impedance analyzer, and mid-sleep time composite phase deviations (CPD) was calculated based on the International Physical Activity Short Questionnaire. Results:SW group had lower serum level of melatonin ( P=0.023) and higher cortisol ( P=0.001) than the NSW group, and altered mRNA expression of Clock and Bmal1 ( P=0.034, P=0.047). Fasting blood glucose and HbA 1C in the SW group, although in the normal range, had been higher than in the NSW group ( P=0.011, P=0.033). Although body mass index was normal in SW group, VFA had been higher than that of the NSW group ( P=0.010). And homeostasis model assessment for insulin resistance (HOMA-IR), IL-6, TNF-α, leptin, chemerin, and visfatin were significantly higher in the SW group than NSW group ( P=0.033, P=0.012, P=0.001, P=0.011, P=0.021, P=0.007). In addition, adjusting for body mass index and activity factors revealed a significant positive correlation between CPD and VFA ( r=0.434, P=0.049), inflammatory factors IL-6 ( r=0.514, P=0.017) and TNF-α ( r=0.700, P<0.001) and pro-inflammatory adipokines leptin ( r=0.473, P=0.030), chemerin ( r=0.439, P=0.047), visfatin ( r=0.521, P=0.015). Conclusion:Shift work can affect circadian clock, with increased visceral adiposity, pro-inflammatory adipokines, inflammatory factors and decreased insulin sensitivity in women without overweight/obese.

Effects of continuous light exposure on muscle fiber remodeling and lipid metabolism in mice / 中华内分泌代谢杂志

Objective:To observe the effects of continuous light exposure on skeletal muscle fiber type transformation and lipid metabolism, and to explore its internal relationship.Methods:Mice were randomly divided into normal light group and 24-hour continuous light group by random number table. The serum and skeletal muscle lipid content and urine 6-sulfatoxymelatonin(6-SML)level were detected by ELISA. The expression of circadian clock and lipid metabolism related genes mRNA were observed by realtime PCR. The muscle fiber type and lipid deposition were evaluated by tissue immunofluorescence as well as oil red O staining.Results:Compared with the normal light group, the level of 6-SML in urine at night decreased( P<0.05), and the expression level and rhythm of brain and muscle ARNT-like protein 1(Bmal1), circadian locomotor output cycles protein kaput(Clock), and period 2(Per2)mRNA in the skeletal muscle changed in continuous light group. In addition, the body weight, blood lipid, free fatty acid, and triglyceride contents of skeletal muscle in continuous light group increased significantly( P<0.05 or P<0.01), the expression of carnitine palmitoyltransferase 1b (Cpt1b)mRNA, the key enzyme of fatty acid oxidation, decreased significantly( P<0.05), while the expression of stearoyl-CoA desaturase(Scd1)mRNA, a lipid synthesis related gene, increased significantly( P<0.01). Further immunofluorescence analysis showed that the proportion of slow muscle fibers decreased and that of fast muscle fibers increased in continuous light group(both P<0.05). Conclusion:The process of ectopic deposition of lipid in skeletal muscle in mice induced by continuous light exposure may be related to the remodeling of skeletal muscle fibers.

The characteristics and clinical significance of serum TRAb IgG subtype in patients with thyroid associated ophthalmopathy / 中华内分泌代谢杂志

Objective:To explore the distribution of serum TRAb IgG subtypes in patients with thyroid-associated ophthalmopathy(TAO) at different stages and its value in assessing TAO activity.Methods:Forty-three patients with TAO, 30 patients with Graves′ disease (GD group), 19 patients with Hashimoto′s thyroiditis (HT group), and 50 healthy subjects (NC group)admitted to the First Affiliated Hospital of Zhengzhou University were collected from August 2018 to February 2019. According to the clinical activity score (CAS), the patients with TAO were further divided into the active period group (AP group, CAS≥3 points, 22 cases) and the inactive period group (IP group, CAS<3 points, 21 cases). The basic clinical data of subjects in each group were collected. The serum concentrations of FT 3, FT 4, TSH, thyroperoxidase antibody (TPOAb), thyroglobulin antibody (TgAb), and thyroid-stimulating hormone receptor antibody (TRAb) were detected by chemiluminescence immunoassay and the binding rate in percentage (B) of serum TRAb IgG and IgG subtypes were detected by ELISA. The positive rate in each group and relative content of positive samples were compared. Results:(1)Compared with HT group, the positive rates of IgG1 and IgG2 in TAO group and GD group were significantly decreased ( P<0.05). There was no significant difference in the relative content of positive TRAb IgG subtype among these disease groups ( P>0.05). (2)Compared with the IP group, IgG1(B) and the positive rates of IgG1 in the AP group were increased while IgG4(B) and the relative contents of IgG4 were reduced ( P<0.05). (3)IgG1(B)was positively correlated with TAO activity ( B=6.190, P=0.007), and higher IgG4(B)indicated more inclinations to the inactive period ( B=-16.390, P=0.052). (4) The area under the curve of receiver operating characteristic (ROC) for TAO developing into active period assessed by activity rate was 0.859 (95% CI 0.746-0.973, P<0.05). When the activity rate was 4.29, the Jordon index showed the largest, with sensitivity of 81.8% and specificity of 81.0%. Conclusions:Elevated levels of TRAb IgG1 in the patients with TAO indicate a tendency to active period, while elevated levels of TRAb IgG4 indicate a tendency to inactive period. The activity rate can provide a reference for assessing whether TAO is active or not.

Clinical significance of combined detection of PD-L1 protein and CD8 +TILs in resected lung adenocarcinoma / 实用肿瘤学杂志

Objective The prognostic role of PD-L1 expression in surgically resected lung adenocarcinoma(ADC)remains controversial. The aim of present study was to investigate the prognostic value of PD-L1 combined with CD8 +TILs expression in pa-tients with resectable lung adenocarcinoma. Methods A total of 104 patients with lung adenocarcinoma who underwent radical resec-tion were enrolled at the Affiliated Tumor Hospital of Harbin Medical University from January 2009 to December 2012. Immunohisto-chemistry was used to detect the expression of PD-L1 and CD8 +TILs in primary tumor specimens. Its clinical pathological factors and its relationship with prognosis were statistical analysis. Results The positive expression rate of PD -L1 in tumor cells was 38. 5% (40/104),and the positive rate of CD8 +TILs was 44. 2% (46/104). There was a significant correlation between the expres-sion of CD8 +TILs and TNM stages of patients with resectable ADC. The Kaplan-Meier survival analysis showed that low expression of PD-L1 and high expression of CD8 +TILs were significantly associated with disease -free survival ( DFS) and overall survival ( OS),and could be used as an independent predictors for predicting prognosis of patients with resectable ADC. Conclusion The ex-pression of PD-L1 and CD8 +TILs in the postoperative specimens can help to judge the prognosis of patients. This study has certain significance for immunotherapy of PD-1/PD-L1 pathway in patients with surgically resected ADC.

The effect of resveratrol on the expression of adiponectin receptors in intermittent high glucose-cultured rat mesangial cells / 中华内分泌代谢杂志

Objective To study the effects and mechanisms of resveratrol (Resv) on the expression of adiponectin receptors ( AdipoR1 and AdipoR2 ) in rat mesangial cells (MCs) cultured in intermittent high glucose medium.Methods The MCs cultured in normal glucose ( 5.6 mmol/L) medium were transfected by the plasmid vector of the FoxO1 short hairpin RNAs ( FoxO1 shRNA ),and then cultured in intermittent high glucose medium (5.6mmol/L or 30 mmol/L,alternately every 8 hours ) and resveratrol ( Resv 20 μmol/L).The MCs cultured in normal glucose (5.6 mmol/L) medium served as normal glucose control group (NG),and the MCs exposed to iutermittent high glucose medium were divided into five groups:intermittent high glucose group (IHG),IHG+Resv group,IHG+FoxO1 shRNA group,IHG+ Resv + FoxO1 shRNA group,IHG + Negative control group ( shNC ),each group was cultured for 72 h.The level of reactive oxygen species (ROS) was assessed by Fluorescence microplate reader.The mRNA levels of Sirt1,Foxo1,AdipoR1,and AdipoR2 were assessed by RT-RCR.The protein levels of FoxO1,phosphorylation FoxO1 ( p-FoxO1 ),AdipoR1,and AdipoR2 were assessed by Western blotting.Results ( 1 )Compared with NG,intermittent high glucose significantly decreased the mRNA expression of Sirt1,but markedly increased the level of p-FoxO1.Furthermore,the mRNA and protein expression levels of AdipoR1 were obviously decreased while the level of ROS was enhanced in IHG ( all P＜0.05 ).(2) Compared with IHG,the mRNA and protein expression levels of FoxO1 and AdipoR1 were inhibited,but the level of ROS was enhanced in group IHG+FoxO1 shRNA ( all P＜0.05 ).( 3 ) Compared with IHG,after treating with Resv,the mRNA expression level of Sirtl was increased,whereas the level of p-FoxO1 was decreased.Moreover,the mRNA and protein levels of AdipoR1 was increased while the level of ROS was lowered in group IHG+Resv (all P＜0.05 ).(4) Compared with group IHG+Resv,the mRNA and protein levels of FoxO1 and AdipoR1 were inhibited,but the level of ROS was increased in group IHG+Resv+FoxO1 shRNA (all P＜O.05).(5) In addition,the mRNA and protein expression level of AdipoR2 showed no significant difference among these groups ( P＞0.05 ).Conclusion ( 1 ) Resveratrol significantly increases the expression of AdipoR1 in MCs induced by intermittent high glucose.( 2 ) FoxO1 plays an important role in regulating the expression of AdipoR1 by resveratrol.

Improvement of mouse spermatogonial cells transplantation method / 解剖学报

Objective To improve the current method of spermatogonial cells transplantation, and make the new method become more operative and easy to carry out. Methods The spermatogonial cells in rC57BL6/tg14 (act-EGFO-Osb Y01) mice that expresse GFP protein were collected as the donor cells, and by using a modified syringe, and then were injected into the seminiferous tubules of pretreated wild type GFP-null mice through testicular efferent duct. The transplantation outcome was evaluated by trypan blue stainting and fluorescent microscopic examination and pathohistological analysis of transplanted testis. Results The transplanted testis of recipient mice showed green fluorescence signal, and the signal of seminiferous tubules was found significantly higher than that of the surrounding tissues. The transplanted GFP-positive cells generated colonies and spermatogenesis but pretreated wild type GFP-null mice were not found. Conclusion The expressed GFP protein spermatogonial cells in rC57BL6/tg14 mice were successfully transplanted in the wild type GFP-null recipient mice, fourthermore the improved transplantation method simplified the triditional one and achieved the same transplantation results.

Molecular imaging of human breast cancer xenografts in nude mice by epidermal growth factor labelled by near infrared fluorescent dye / 中华放射学杂志

can specifically combined with EGFR, which may be applied to noninvasive NIRF imaging of tumors highly expressed EGFR in vivo.