ACTA ENDOCRINOLOGICA (BUC)

Objective. To observe the impact of quercetin and isoquercitrin on gluconeogenesis in hepatocytes. Methods. Mouse primary hepatocytes were cultured with lactic acid and pyruvic acid. After treatment with quercetin and isoquercitrin for 24 hours, the glucose concentration in the culture supernatant was determined. RT-PCR was used to detect the mRNAs of PEPCK, G6Pase, LKB1, and AMPKα. Protein levels of LKB1, AMPKα, and Thr172 phosphorylation were evaluated by Western blot. Results. The glucose concentration in the gluconeogenesis group (GN) was significantly higher than in the control group (C), but the glucose concentrations in the high level quercetin(group 80Q) and high level isoquercitrin (group 80I) were significantly lower than in the group GN, P<0.01. In the group 80Q, and group 80I, the mRNA levels of PEPCK and LKB1were significantly lower than in the group GN (P<0.01), and the G6Pase mRNA were significantly lower than in the group GN (P<0.05). The protein levels of LKB1 and the phosphorylation of AMPKα Thr172 in the group 80Q, group 40I, and group 80I were higher than in the group GN. The effects of quercetin and isoquercitrin on LKB1 and AMPKα were similar to those of metformin. Conclusions. Quercetin and isoquercitrin inhibit gluconeogenesis in hepatocytes, which may be related to the LKB1 upregulation and phosphorylation of AMPKα.

Objective. This study was to investigate RIP140 expression levels in the pancreas and islet β-cells in mice and rats and the role of RIP140 in cultured β-cells using the mouse pancreatic β-cell line MIN6. Methods. The MIN6 cell line stably overexpressing RIP140 was used. The effects of RIP140 on cell viability, cell cycle, apoptosis, insulin secretion, and its regulated genes were analyzed using flow cytometry, the MTT assay, Western blot analysis, reverse transcriptase (RT)-PCR, and enzyme-linked immunosorbent assay (ELISA). Results. Most of insulin-positive cells in islets expressed RIP140. In MIN6 cells, overexpression of RIP140 inhibited cell viability by reducing the number of cells in S phase and inhibiting proliferating cell nuclear antigen (PCNA) expression. We also found that overexpression of RIP140 inhibited Bcl-2 and mRNA expression of peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α) and uncoupling protein 2 (UCP2) and increased levels of phosphorylated extracellular signalregulated protein kinases 1/2 (p-ERK1/2). However, apoptosis rate and levels of basal level of insulin secretion (BIS) and glucose-stimulated insulin (GSIS) were not significantly altered in MIN6 cells. Conclusions. RIP140 was expressed in the pancreas of mice and rats, particularly in β-cells, and participated in regulating β-cell function and proliferation.

Context. To analyze the dynamic changes of serum thyrotrophin receptor antibody (TRAb) and thyroid peroxidase antibody (TPOAb) in Graves’ disease (GD) patients before and after radioactive iodine (RAI) treatment and to investigate if TRAb and TPOAb play a role in the occurrence of early hypothyroidism after 131I therapy for Graves’ hyperthyroidism. Subjects and Methods. A total of 240 patients newly diagnosed with GD were selected to study. A clinical and laboratory assessment was performed before and at 3, 6, and 12 months after 131I therapy. Chemiluminescent immunoassays were used to detect serum free triiodothyronine (FT3), free thyroxine (FT4), sensitive thyroid-stimulating hormone (TSH) and TPOAb concentration. Radio-receptor assay was used to measure serum TRAb concentration. According to the early onset of hypothyroidism in a year after RAI therapy, patients were divided into early hypothyroidism group (group A) and non-early hypothyroidism group (group B). Results. In both groups, serum TRAb and TPOAb increased at 3 months, reached the highest level at 6 months and returned to the baseline at 12 months after RAI therapy. TRAb showed a significant difference between the two groups at 6 months (P<0.01). Serum TPOAb in group A was higher than that in group B before and at 3, 6, 12 months after RAI therapy (P<0.05). Conclusions. Serum TRAb and TPOAb are closely related to the occurrence of the early hypothyroidism, and play an important role in judging prognosis after 131I treatment in Graves’ disease.

Background. In vitro studies of the changes about osteoblastogenesis and adipogenesis potential of BMSCs were not clear. As it is the critical pathway for osteogenic differentiation and bone formation, whether or not Wnt/β- catenin signalling is involved in the changes of osteogenic and adipogenic potential of BMSCs and participates in bone content decrease of ovariectomized (OVX)osteoporosis rats has been rarely reported. Material/Methods. BMSCs from femurs of ovariectomzed rats were isolated and cultured in vitro. The proliferation potential of BMSCs was analysed by CCK-8 assays . Osteoblastic and adipogenic differentiation potential of the BMSCs was assessed by ALP activity assay, Alizarin red S staining, Oil red O staining and RT-PCR analysis. Results. The results demonstrated that BMSCs from bilateral ovariectomization rats were endowed with lower proliferation and osteoblastic differentiation potential but higher adipogenic potential than the control group in vitro. In addition, β-catenin was found to have been decreased in OVX BMSCs, indicating that Wnt/β-catenin signalling pathways were suppressed in OVX BMSCs . Conclusions. Results suggested that changes in the Wnt canonical signalling pathway may be related to imbalances of osteogenic and adipogenic potential of BMSCs, and this may be an important factor related to bone content decrease in ovariectomized osteoporosis rats.

Background & Aims. Results from observational studies on low carbohydrate and high protein (LCHP) diets on all-cause mortality and cause-specific mortality are inconsistent. The objective of the present meta-analysis was to investigate LCHP diets and mortality, and cause-specific mortality. Methods. Medline, EMBASE and web of science database were searched for cohort studies published from January 2000 to April 2013. A database was developed on the basis of 7 cohorts from 6 studies, including 279,332 individuals with an average 13.2 years of follow-up. Results. A significantly increased risk of all-cause mortality was observed among those who adhere to LCHP diets [(HR (95% CI): 1.06 (1.04, 1.09); p = 0.001; I2 = 8.2% (p = 0.336)]. However, there were no significant associations between LCHP diets and mortalities from cancer [(HR (95% CI): 1.02 (0.98, 1.05); p = 0.097; I2 = 59.9% (p = 0.041)] and CVD [(HR (95% CI): 1.01 (0.98, 1.03); p = 0.428; I2 = 24.0% (p = 0.0.254)]. Furthermore, a significantly decreased risk of all-cause mortality among those who adhere to high carbohydrate diets [(HR (95% CI): 0.97 (0.94, 0.98); p <0.001; I2 = 5.7% (p = 0.364)] was observed. Conclusion. Present results support that long-term LCHP diets could cause increased mortality of all-cause, while high carbohydrate diets have a protective effect for all-cause mortality. Our analysis provides health professionals with an incentive to pay attention to the adverse effects of LCHP diets on health.

Objectives. To assess the effects of electroacupuncture (EA) at the Zusanli (ST36), Guanyuan (CV4), Zhongwan (CV12), and Fenglong (ST40) acupoints on sirtuin 1 (SIRT1) and glucose transporter type 4 (GLUT4) expression in high-fat diet (HFD)-induced insulin-resistant (IR) rats. Methods. Wistar rats were divided into normal control (NC), HFD, and HFD+EA groups. NC rats were fed a standard chow diet and did not receive EA. After being fed an HFD for eight weeks, rats in the HFD+EA group received EA at 2 Hz five times a week for eight weeks. Rats in the HFD group did not receive EA. Results. In HFD-induced IR rats, EA inhibited body weight increase and water intake, which were observed in HFD rats. EA had no effect on fasting blood glucose and postprandial blood sugar levels. Intraperitoneal insulin tolerance testing revealed that EA enhanced insulin sensitivity in HFD-induced IR rats. Compared with NC rats, SIRT1 and GLUT4 were downregulated in the quadriceps femoris of HFD-fed rats but were increased after eight weeks of EA stimulation. Conclusions. EA enhanced HFD-induced insulin resistance by activating SIRT1 and GLUT4 in the quadriceps femoris. These results provide powerful evidence supporting the beneficial effects of EA on HFD-induced insulin resistance.

Context. The 131I activity for treating Graves’ disease (GD) is usually determined based on physician’s experience. Objective. This study aimed to design an empirical method that was not only personalized and quantitative, but also simple, convenient, and easy to grasp. Subjects and Methods. The study population comprised patients with GD, selected between May 2013 and May 2016, who received 131I therapy in the Outpatient Department of Shanghai Ninth People’s Hospital. The firstvisit patients of physician 1 were placed in the traditional group: the activity of 131I (mCi) was calculated using the routine formula: [empirical activity (0.07–0.12 mCi/g) × thyroid mass]/[24-h thyroid 131I uptake]. The first-visit patients of physician 2 were placed in the personalized group. The activity of 131I (mCi) was calculated in two steps. First, the initial activity was calculated: 0.1 mCi/g × thyroid mass (g), and then a personalized and quantitative calibration table of 131I activity was used to obtain a final 131I activity. The cure rate with a single activity of 131I was recorded 1 year later. Results. The traditional and personalized groups included 241 and 282 patients, respectively. Interestingly, the personalized group achieved a higher cure rate [86.5% (244/282) versus 73.4% (177/241), P = 0.000] with a relatively higher 131I activity for the first treatment [8.7 (7, 3.5-30) mCi versus 6.7(6, 2.5-30) mCi, P = 0.000] compared with the traditional group, while the incidence rate of permanent hypothyroidism was not significantly different between the two groups (P = 0.175). Conclusion. The empirical method designed in this study was reliable.

Aim. To establish a nomogram combining preoperative ultrasonic and clinical features for predicting lymph nodes posterior to the right recurrent laryngeal nerve (LN-prRLN) metastasis in papillary thyroid carcinoma (PTC) patients. Methods. Preoperative ultrasonic and clinical variables of patients with PTC from 2014 to 2021 were retrospectively analyzed. The risk factors associated with LN-prRLN metastasis were identified and validated through a developed nomogram model based on univariate and multivariate logistic regression analysis. Results. A total of 615 patients (690 lesions) were enrolled for the training dataset and 207 patients (226 lesions) for the validation dataset with 54 (6.57%) patients developing LN-prRLN metastasis. Multivariate logistic regression analysis demonstrated that the preoperative ultrasound measurement of larger tumors (≥20 mm), higher TI-RADS category (category 5), and higher thyroglobulin level (9.86 ng/mL) in patients with PTC were predictive factors for LN-prRLN metastasis. The nomogram model was established and verified yielding a relatively good predictive performance in the training and validation dataset (AUC: 0.868 vs. 0.851). Conclusions. The nomogram combining preoperative ultrasonography with clinical features in this study is highly predictive of LN-prRLN metastasis in patients with PTC, which may provide more personalized recommendations for clinicians in preoperative decisionmaking for complete dissection of LN-prRLN.

Context. Diabetic peripheral neuropathy (DPN) is a common complication associated with long-term type 2 diabetes mellitus, although early diagnosis can improve prognosis. Objective. Our objective was to develop a simple protocol for early diagnosis of DPN in Chinese type 2 diabetic patients. Subjects and Methods. A total of 209 type 2 diabetic patients were included; these patients were categorized as symptomatic and asymptomatic group based on their symptoms. Clinical data of these patients were recorded and they were screened for DPN by vibration perception threshold test (VPT), 10-G nylon monofilament test, temperature identification, and the tendon reflex test. Results. The total combined rate of patients who were tested positive for DPN with all four screening methods was 68.7%. Patients tested positive for DPN were significantly older and had a longer disease duration than those who were tested negative (p<0.01); however, glycated hemoglobin levels, presence of hypertension, and gender did not differ significantly between them (p>0.05). Among screening methods, the highest positive rate observed among patients screened with the VPT test was 63.64% as compared to other tests. The total positive rate for temperature discrimination, 10-G monofilament and tendon reflex test were 26.79%, 11.96 % and 17.22 % respectively. In asymptomatic group VPT showed the highest positive rate for DPN (48.41%). Conclusions. The combination of four simple methods can improve the detection rate of DPN and identify subclinical cases. Abnormal vibration perception was the most common feature of DPN and it was associated with both disease duration and the age of the patient.

Cimicifuga racemosa (CR) is widely used in the treatment of menopausal symptoms, including hormone-related affective disorders in women. Mechanistic studies suggest that unlike hormone-replacement therapy, CR does not stimulate cancerous growth.\r\nObjective: to evaluate CR safety and the antidepressant-like effects compared to estrogen treatment.\r\nMaterials and Methods: we used the forced swimming test (FST) to address the hypotheses that CR alters behavior of ovariectomized (OVX) rats in the FST and the FST-induced expression of c-fos in the rat paraventricular nucleus (PVN). After the FST, all animals were perfused, and the brains were processed for c-fos immunocytochemistry; the plasma was processed for radioimmunoassay of plasma estrogen levels.\r\nResults: OVX group animals spent significantly (P<0.05) more time struggling than CR and E2 groups. The CR and E2 groups animals spent little time in struggle time decreased. CR and E2 groups animals spent significantly (P<0.05) more time swimming than OVX group animals.\r\nConclusion: CR was demonstrated to have antidepressant effect, and establishing the safety symptoms in women in whom hormone-replacement therapy is contraindicated.