Ameliorating effect of Trigonella foenum-graecum L. (fenugreek) extract tablet on exhaustive exercise-induced fatigue in rats by suppressing mitophagy in skeletal muscle.

OBJECTIVE: Trigonella foenum-graecum L. (fenugreek) is widely used as a leafy vegetable and spice in China and North Africa. Recent studies have reported that fenugreek can reduce fatigue; however, its antifatigue mechanism remains unclear. Therefore, this study aimed to investigate the potential antifatigue effects of fenugreek extract (FE) on mitophagy and the underlying mechanisms. MATERIALS AND METHODS: We evaluated the potential effects of FE tablet on an exhaustive exercise-induced fatigue (EEF) rat model. Oxidative stress indicators and fatigue biomarkers in the serum and skeletal muscle were detected. Mitophagy and mitochondrial morphology were observed using transmission electron microscopy. The expression levels of mitochondrial autophagy-related proteins were detected using western blot and immunofluorescence. RESULTS: Compared with the model group, FE enhanced the activities of the antioxidant enzymes superoxide dismutase and glutathione peroxidase as well as total antioxidant capacity; however, it decreased the level of malondialdehyde in the serum and skeletal muscle after a 7-day treatment. Moreover, certain indicators of mitochondrial function, such as reactive oxygen species levels, ATP levels, cellular and mitochondrial Ca2+ levels, and ATPase activity, were significantly improved in the FE group compared with the model group. Finally, we found that mitophagy was induced by exhaustive exercise and inhibited by FE. Regarding mitochondrial autophagy-related proteins, the expression levels of LC3B, FUNDC1, PGAM5, PARKIN, and PINK1 in the skeletal muscle tissue were increased in the EEF group compared with the control group. After administration of FE and a positive control drug, a significant reversal in the expression of the above-mentioned proteins was noted. CONCLUSIONS: Our findings demonstrate that FE exerted antifatigue effects in the EEF rat model by regulating the mitophagy-related FUNDC1/LC3B signaling pathway rather than the PINK1/PARKIN signaling pathway.

[Establishment of an auxiliary diagnosis system of newborn screening for inherited metabolic diseases based on artificial intelligence technology and a clinical trial].

Objective: To establish a disease risk prediction model for the newborn screening system of inherited metabolic diseases by artificial intelligence technology. Methods: This was a retrospectively study. Newborn screening data (n=5 907 547) from February 2010 to May 2019 from 31 hospitals in China and verified data (n=3 028) from 34 hospitals of the same period were collected to establish the artificial intelligence model for the prediction of inherited metabolic diseases in neonates. The validity of the artificial intelligence disease risk prediction model was verified by 360 814 newborns' screening data from January 2018 to September 2018 through a single-blind experiment. The effectiveness of the artificial intelligence disease risk prediction model was verified by comparing the detection rate of clinically confirmed cases, the positive rate of initial screening and the positive predictive value between the clinicians and the artificial intelligence prediction model of inherited metabolic diseases. Results: A total of 3 665 697 newborns' screening data were collected including 3 019 cases' positive data to establish the 16 artificial intelligence models for 32 inherited metabolic diseases. The single-blind experiment (n=360 814) showed that 45 clinically diagnosed infants were detected by both artificial intelligence model and clinicians. A total of 2 684 cases were positive in tandem mass spectrometry screening and 1 694 cases were with high risk in artificial intelligence prediction model of inherited metabolic diseases, with the positive rates of tandem 0.74% (2 684/360 814)and 0.46% (1 694/360 814), respectively. Compared to clinicians, the positive rate of newborns was reduced by 36.89% (990/2 684) after the application of the artificial intelligence model, and the positive predictive values of clinicians and artificial intelligence prediction model of inherited metabolic diseases were 1.68% (45/2 684) and 2.66% (45/1 694) respectively. Conclusion: An accurate, fast, and the lower false positive rate auxiliary diagnosis system for neonatal inherited metabolic diseases by artificial intelligence technology has been established, which may have an important clinical value.

CircRNA ZNF609 promotes growth and metastasis of nasopharyngeal carcinoma by competing with microRNA-150-5p.

OBJECTIVE: This study aims to explore the biological function of circular RNA ZNF609 (circ-ZNF609) in regulating the occurrence and progression of nasopharyngeal carcinoma (NPC), and to investigate the possible underlying mechanism. PATIENTS AND METHODS: The expression levels of circ-ZNF609, microRNA-150-5p and Sp1 in NPC tissues and normal nasopharyngeal epithelial tissues were detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Circ-ZNF609 expression was silenced in NPC cell lines (5-8F and HONE-1). Cellular behaviors of NPC cells were determined using Cell Counting Kit-8 (CCK-8), wound healing and transwell assay. The binding relationship among microRNA-150-5p, circ-ZNF609 and Sp1 was detected by Dual-Luciferase reporter gene assay. In addition, the protein expression of Sp1 after altering expression of circ-ZNF609 or microRNA-150-5p was detected by Western blot. RESULTS: The expression levels of circ-ZNF609 and Sp1 in NPC tissues were markedly higher than those of normal nasopharyngeal epithelial tissues. However, the expression of microRNA-150-5p was significantly lower in NPC tissues. The knockdown of circ-ZNF609 in NPC cells 5-8F and HONE-1 significantly inhibited the proliferative, migratory and invasive behaviors of NPC cells. Meanwhile, microRNA-150-5p knockdown in NPC cells showed the opposite effect on cellular behaviors of NPC cells. Dual-Luciferase reporter gene assay revealed that circ-ZNF609 could bind to microRNA-150-5p, and Sp1 was a target gene of microRNA-150-5p. Western blot results showed that circ-ZNF609 could stabilize the expression of Sp1, while microRNA-150-5p degraded Sp1 expression. Furthermore, the knockdown of Sp1 in NPC cells reversed the carcinogenic effect of circ-ZNF609. CONCLUSIONS: Highly expressed circ-ZNF609 adsorbs microRNA-150-5p to upregulate Sp1 expression, thereby promoting the proliferation and metastatic ability of NPC cells.

Risk of bone fracture associated with sodium-glucose cotransporter-2 inhibitor treatment: A meta-analysis of randomized controlled trials.

AIM: To evaluate the association between sodium-glucose cotransporter-2 (SGLT2) inhibitors and risk of bone fractures in patients with type 2 diabetes mellitus (T2DM). METHODS: A systematic literature search conducted of PubMed, Embase, the Cochrane Library and Web of Science from inception up to 31 August 2018 identified all eligible randomized controlled trials (RCTs). The following data were extracted from each study: first author; year of publication; sample size; patient characteristics; study design; intervention drug; control drug; follow-up durations; and incident bone-fracture events. A meta-analysis was performed using Review Manager 5.3 software to calculate odds ratios (ORs) and 95% confidence intervals (CI) for dichotomous variables. RESULTS: A total of 30 studies involving 23,372 patients with T2DM were included in our analysis. There were 387 incident bone-fracture cases (245 in the SGLT2 inhibitor group, 142 in the control group). Compared with patients who received placebo, those receiving SGLT2 inhibitor treatment had a pooled OR of bone fracture of 0.86 (95% CI: 0.70-1.06). Also, there was no evidence that individual SGLT2 inhibitors across different doses were associated with any increased risk of bone fracture. After stratification by follow-up duration, an SGLT2 inhibitor treatment period of ≤ 52 weeks appeared to have beneficial effects against bone fracture; however, when the treatment period exceeded 52 weeks, these beneficial effects for preventing bone fracture disappeared. CONCLUSION: Our meta-analysis has indicated that SGLT2 inhibitors do not increase risk of bone fracture compared with placebo in patients with T2DM. However, these findings now need to be confirmed in well-designed RCT studies.

[Efficacy and safety of endonasal phototherapy in the treatment of adult allergic rhinitis: A Meta-analysis].

Objective:To perform a systematic review of the efficacy and safety of endonasal phototherapy in the treatment of AR, and to provide evidence of evidence-based medicine for clinical application. Method:Databases were from PubMed, Embase, Cochrane Library, Chinese Academic Journal Full-text Database (CNKI) and Wanfang database. A randomized controlled trial (RCT) or clinical study literature on intranasal phototherapy for adult AR is available, and the deadline is March 2017. Based on the literature inclusion and exclusion criteria, the related literatures were selected and the quality was evaluated by using the Cochrane inclusion bias assessment table. Meta-analysis was performed with Revman 5.3 software. For continuous outcomes, the weighted mean difference (WMD) and its 95% confidence intervals (CI) were calculated, forest maps and funnel plots were drew. For uncontinuous outcomes, the odds ratio (OR) and its 95%CI were calculated, and forest maps and funnel plots were drew. The efficacy included total nasal symptom scores (TNSS), rhinoconjunctivitis quality of life questionnaire (RQLQ) and palate itching scores, and the safety was assessed by incidence rate of severe drying and mild drying of nasal mucosa. Result:A total of 12 articles were selected, including 5 RCT and 7 clinical studies, and there were 615 adult AR patients. The Meta-analysis shows that endonasal phototherapy significantly reduced the TNSS, RQLQ and palate itching scores, the incidence rate of mild drying of nasal mucosa was higher in comparison with pretreated values, and the difference was statistically significant (P<0.05). The rate of severe drying of nasal mucosa was higher, but the difference was not statistically significant (P>0.05). It also shows that endonasal phototherapy significantly reduced the TNSS, RQLQ and palate itching scores, and the rate of mild drying of nasal mucosa was higher in comparison with placebo and antihistamines groups, and the difference was statistically significant. The rate of severe drying of nasal mucosa was higher, but the difference was not statistically significant. Conclusion:Endonasal phototherapy can improve the symptoms of TNSS, RQLQ and palate itching score in patients, and the safety is also confirmed.Patients who are not satisfied with symptom relief may choose to use it.

Hepatic dysfunction related to thyrotropin receptor antibody in patients with Graves' disease.

BACKGROUND: Hepatic dysfunction is a common phenomenon in patients with Graves' disease (GD). However, its pathogenesis is not fully understood. We aimed to determine the correlation between the thyrotropin receptor antibody (TRAb) and liver biochemical abnormalities in patients with GD. METHODS: A total of 236 consecutive unrelated inpatients with newly diagnosed and untreated GD were included. Clinical characteristics (age, gender, disease duration) were collected. The liver biochemical values were tested and serum thyroid hormones, anti-thyroid antibodies and thyroid volumes were also evaluated. The patients were divided into hepatic dysfunction (HDF) and normal hepatic function (NHF) groups according to liver biochemical values. RESULTS: We found that 77.9% untreated patients with GD had at least one liver function test abnormality. The levels of TRAb in patients of HDF group were significantly increased compared with those in patients of NHF group, P < 0.001. Linear regression suggested that TRAb has significant correlation with AST, ALP, γ-GTP, TB and DB. Logistic regression concluded that GD patients with high levels of TRAb had a greater possibility of developing liver biochemical abnormalities (OR = 1.069, 95% CI 1.019-1.113). CONCLUSIONS: Hepatic dysfunction is common in patients with GD, and elevation of TRAb may contribute to hepatic dysfunction in patients with GD.

Genome-wide analysis of DNA methylation variations caused by chronic glucolipotoxicity in beta-cells.

There is a growing body of literature suggesting the role of interactions between genes and the environment in development of type 2 diabetes mellitus (T2DM). However, the interplay between environment and genetic in developing and progressing T2MD is not fully understood. To determine the effects of high-glucose-lipid on the status of DNA methylation in beta cells, and clarify the mechanism of glucolipotoxicity on beta-cell deterioration, the DNA methylation profile was detected in beta-cells cultured with high-glucose-lipid medium.We utilized a high throughput NimbleGen RN34 CpG Island & Promoter Microarray to investigate the DNA methylation profile in beta-cells cultured with high-glucose-lipid medium. To validate the results of microarray, the immunoprecipitation (MeDIP) PCR was used to test the methylation status of some selected genes. The mRNA and protein expression of insulin and Tcf7l2 in these cells were quantified by RT-PCR and western blot, respectively.We have identified a lot of loci which experienced aberrant DNA methylation in beta-cells cultured with high-glucose-lipid medium. The results of MeDIP PCR were consistency to the microarray. An opposite regulation in transcription and translation of Tcf7l2 gene was found. Furthermore, the insulin mRNA and protein expression in beta-cells also decreased after cultured with high-glucose-lipid medium compared with the control cells.We conclude that chronic glucolipotoxicity could induce aberrant DNA methylation of some genes and may affect these genes expression in beta-cells, which might contribute to beta-cell function failure in T2DM and be helpful to explain, at least partially, the mechanism of glucolipotoxicity on beta-cells deterioration.

[Effect of pulse current iontophoretic transdermal delivery of insulin on blood glucose in diabetic rats].

Iontophoresis of pulse current with various current intensity, frequency, on/off ratio and duration of treatment was used to facilitate the transdermal delivery of insulin in order to control blood glucose levels in diabetic rats. Male Sprague-Dawley rats were made diabetic by i.p. injection of streptozotocin (65 mg . kg-1 body weight). After two days, diabetic rats were anaesthetized with urethane (as 25% aqueous solution, 1.5 mg . g-1). Both the reservoir electrodes and the receptor electrodes were applied on the abdominal site of the diabetic rats. Iontophoresis was carried out using a prototype transdermal periodic iontophoretic system in order to provide the required direct current with desired pulse modes. The extent of reduction of blood glucose levels was found to be positively correlated with the current intensity, frequency and duration of treatment to some extent. But when the current intensity was over 0.8 mA/cm2 and the frequency was over 3000 Hz, the reduction of blood glucose levels did not continuously increase. Blood glucose levels were found to be better controlled when the on/off ratio of 1 : 1 was used.

[Facilitated transdermal delivery of insulin by pulse current iontophoresis].

Facilitated transdermal delivery of insulin by pulse current iontophoresis was investigated. It was found that pulse current iontophoresis can increase the transdermal permeation rate, and there was a positive correlation between reservoir insulin concentration and skin permeation rate of insulin. Moreover, when the reservoir solution pH (3.6) was below the isoelectric point of insulin (pI 5.2), the transdermal permeation rate of insulin was the highest, 324.3 +/- 33.4 microU/(cm2.h). When the pH of the reservoir solution was brought up to 7.4, the transdermal permeation rate of insulin declined markedly to 143.7 +/- 27.3 microU/(cm2.h). When the reservoir solution pH was close to the isoelectric point of insulin, the transdermal permeation rate of insulin was the lowest, 78.4 +/- 21.9 microU/(cm2.h).

[Effects of silybin on red blood cell sorbitol and nerve conduction velocity in diabetic patients].

The effects of silybin on red blood cell (RBC) sorbitol and nerve conduction velocity in 14 non-insulin dependent diabetic patients (female 9, male 5; average age 58.2 years) were reported. Their RBC sorbitol levels averaged 72.55 +/- 21.61 nmol/g.Hb, a value almost two times of non-diabetic controls (33.31 +/- 7.82 nmol/g.Hb). After 4 weeks of silybin (231 mg/d) therapy, RBC sorbitol dropped to 39.53 +/- 14.94 nmol/g.Hb, a highly significant reduction than that before silybin therapy. Silybin treatment had no effect on fasting blood glucose. In addition, silybin treatment slightly improved nerve conduction velocity, but statistically not significant. This report suggests that silybin may be a potent aldose reductase inhibitor, and valuable in the prophylaxis and treatment of diabetic complications.

Effects of nicotinamide on cardiac contraction force and slow inward current.

The effects of nicotinamide (Nic) on cardiac contraction force and on slow inward current (Isi) in guinea pigs were studied with atrial strips and voltage clamp techniques. Verapamil (Ver), MnCl2, nifedipine (Nif), and propranolol (Pro) depressed markedly the positive inotropic effect induced by Nic in a noncompetitive manner. The pD2 values of Ver, MnCl2, Nif, and Pro were 6.19, 3.41, 5.00, and 6.43, respectively. The action of Nic was reduced by a low Ca2+ Tyrode solution and disappeared in a Ca(2+)-free solution. Nic 33 mmol.L-1 elevated the Isi from 6.5 +/- 1.3 microA to 10.3 +/- 2.2 microA. The results suggest that Nic promotes the Ca2+ influx and its site of action is different from that of both Pro and the calcium antagonists.

[Inhibition of aldose reductase by Chinese herbal medicine].

Seven Chinese herbal drugs were screened for experimental inhibition of lens aldose reductase activity, among which quercetin exhibited potent enzyme-inhibitory activities in vitro. Its IC50 value was 3.44 x 10(-7) mol/L. It may be helpful in the prophylaxis and treatment of diabetic complications.

[Effects of 2[p-(dimethylamino)styryl] pyridine methiodide on mouse and rabbit].

The effects of 2[p-(dimethylamino)styryl] pyridine methiodide on mouse and rabbit ECG and on the contraction of isolated rabbit atrial muscles were studied. DSPM produced long-lasting bradycardia and A-V block in a dose-dependent manner, but did not effect intraventricular conduction. DSPM antagonized the positive chronotropic and dromotropic effects of isoprenaline (Iso) in vivo, and antagonized the inotropic effects of Iso and CaCl2 non-competitively in vitro. The pD'2 were 4.49 and 4.52, respectively. It is suggested that DSPM may be a Ca2+ antagonist.