Liraglutide improved the reproductive function of obese mice by upregulating the testicular AC3/cAMP/PKA pathway.

BACKGROUND: The incidence of male reproductive dysfunction is increasing annually, and many studies have shown that obesity can cause severe harm to male reproductive function. The mechanism of male reproductive dysfunction caused by obesity is unclear, and there is no ideal treatment. Identification of effective therapeutic drugs and elucidation of the molecular mechanism involved in male reproductive health are meaningful. In this study, we investigated the effects of the GLP-1 receptor agonist liraglutide on sex hormones, semen quality, and testicular AC3/cAMP/PKA levels in high-fat-diet-induced obese mice. METHODS: Obese mice and their lean littermates were treated with liraglutide or saline for 12 weeks. Body weight was measured weekly. Fasting blood glucose (FBG) was measured using a blood glucose test strip. The serum levels of insulin (INS), luteinizing hormone (LH), follicle stimulating hormone (FSH), testosterone (T), free testosterone (F-TESTO), estradiol (E2), and sex hormone binding globulin (SHBG) were detected using ELISA. The sperm morphology and sperm count were observed after Pap staining. The mRNA and protein expression levels of testicular GLP-1R and AC3 were measured by RT-qPCR and Western blot, respectively. Testicular cAMP levels and PKA activity were detected using ELISA. RESULTS: Liraglutide treatment can decrease body weight, FBG, INS, HOMA-IR, E2 and SHBG levels; increase LH, FSH, T, and F-TESTO levels; increase sperm count; decrease the sperm abnormality rate; and increase GLP-1R and AC3 expression levels and cAMP levels and PKA activity in testicular tissue. CONCLUSIONS: Liraglutide can improve the sex hormone levels and semen quality of obese male mice. In addition to its weight loss effect, liraglutide can improve the reproductive function of obese male mice, which may also be related to the upregulation of AC3/cAMP/PKA pathway in the testis. This work lays the groundwork for future clinical studies.

Evidence basis for using dexmedetomidine to enhance the quality of paravertebral block: A systematic review and meta-analysis of randomized controlled trials.

Background: Dexmedetomidine is considered an adjunct to local anaesthesia (LA) to prolong peripheral nerve block time. However, the results from a previous meta-analysis were not sufficient to support its use in paravertebral block (PVB). Therefore, we performed an updated meta-analysis to evaluate the efficacy of dexmedetomidine combined with LA in PVB. Methods: We performed an electronic database search from the date of establishment to April 2022. Randomized controlled trials (RCTs) investigating the combination of dexmedetomidine and LA compared with LA alone for PVB in adult patients were included. Postoperative pain scores, analgesic consumption, and adverse reactions were analyzed. Results: We identified 12 trials (701 patients) and found that the application of dexmedetomidine as a PVB adjunct reduced the postoperative pain severity of patients 12 and 24 h after surgery compared to a control group. Expressed as mean difference (MD) (95% CI), the results were -1.03 (-1.18, -0.88) (p < 0.00001, I2 = 79%) for 12 h and -1.08 (-1.24, -0.92) (p < 0.00001, I2 = 72%) for 24 h. Dexmedetomidine prolonged the duration of analgesia by at least 173.27 min (115.61, 230.93) (p < 0.00001, I2 = 81%) and reduced postoperative oral morphine consumption by 18.01 mg (-22.10, 13.92) (p < 0.00001, I2 = 19%). We also found no statistically significant differences in hemodynamic complications between the two groups. According to the GRADE system, we found that the level of evidence for postoperative pain scores at 12 and 24 h was rated as moderate. Conclusion: Our study shows that dexmedetomidine as an adjunct to LA improves the postoperative pain severity of patients after surgery and prolongs the duration of analgesia in PVB without increasing the incidence of adverse effects.

Upregulation of circ-0000745 in acute lymphoblastic leukemia enhanced cell proliferation by activating ERK pathway.

Acute lymphoblastic leukemia (ALL) is a common hematologic malignancy. Circular RNAs (circRNAs) have been reported as an important factor in regulating cellular process expressed in all hematopoietic compartment. But the molecular mechanism of circRNAs in ALL remain unclear. In this study, we observed circ-0000745 upregulated in leukemia cells (Kasumi-1 and KG-1). Upregulated the expression of circ-0000745 significantly enhanced the proliferation of Kasumi-1 and KG-1 cells, and reduced the ratio of apoptosis cells. Knock down the endogenous expression of circ-0000745 suppressed the cell proliferation and increased the ratio of apoptosis cells. Furthermore, western blot assay showed that overexpression of circ-0000745 increased the activity of ERK1/2. Altogether, these results revealed that upregulated circ-0000745 in leukemia cells could promoter cell viability by increasing the activation of ERK pathway. This study supported the important of circRNAs in the process of acute lymphoblastic leukemia, and provided a new biomarker on ALL diagnosis and therapy.

Stable knockdown of ZBTB7A promotes cell proliferation and progression in nasopharyngeal carcinoma.

AIMS: Although high expression of ZBTB7A is positively relative to metastasis in nasopharyngeal carcinoma (NPC) patients, the association between its low expression and metastasis of NPC remains unclear. The present study aimed to definitely identify the association. METHODS: The level of ZBTB7A was effectively knocked down by stable transfection of short hair RNA plasmid in NPC cell lines CNE2 and 5-8F (shRNA-CNE2 and shRNA-5-8F), compared with the cells that stably transfected empty plasmid (NC-CNE2 and NC-5-8F). The levels of ZBTB7A were assessed by real-time polymerase chain reaction and Western blot in the cell lines. MTT assay, colorimetric focus-formation assay, flow cytometry, wound healing assay, transwell assays, and xenograft model were performed to analyze cell vitality, proliferation, cell cycle, migration, invasion, and tumorigenicity. RESULTS: The levels of ZBTB7A were effectively reduced in shRNA-CNE2 and shRNA-5-8F. Their carcinogenicity was stronger separately than the abilities of NC-CNE2 and NC-5-8F. NC-CNE2 and shRNA-CNE2 were selected to establish the xenograft model because of their stronger tumorigenicity than NC-6-10B and shRNA-5-8F. The assay showed that shRNA-CNE2 had stronger tumorigenicity than NC-CNE2. CONCLUSIONS: The results demonstrated the reverse association between the expression of ZBTB7A and the tumorigenicity of NPC. We postulate that some oncogenic pathways, which are suppressed by ZBTB7A, will vicariously promote the proliferation and progression of NPC when ZBTB7A is decreased.

Circulating chemerin levels elevated in dilated cardiomyopathy patients with overt heart failure.

BACKGROUND: Recent evidence demonstrated that the circulating concentrations of adipokine are related to the presence of heart failure secondary to ischemic heart disease and dilated cardiomyopathy (DCM). However, the plasma concentrations of chemerin in patients with DCM have yet to be investigated. METHODS: The present study enrolled 109 DCM patients with typical symptoms of heart failure and 60 healthy controls and measured plasma concentrations of chemerin, IL-6 and TNF-α using enzyme-linked immunosorbent assay. Left ventricular end-diastolic diameter (LVEDD) and left ventricular ejection fraction (LVEF) were measured using a GE ViVid E7 ultrasonography machine. RESULTS: Plasma chemerin, IL-6 and TNF-α concentrations were significantly higher in DCM patients compared to the control group. A correlation analysis revealed that plasma chemerin concentrations were positively correlated with the concentrations of IL-6 (R=0.270, P=0.004), TNF-α (R=0.302, P=0.001), C-reactive protein (CRP) (R=0.256, P=0.004), N-terminal pro-brain natriuretic peptide (NT-proBNP) (R=0.386, P=0.000), and LVEDD (R=0.212, P=0.027) but negatively correlated with LVEF (R=-0.543, P=0.000). Furthermore, chemerin (OR 1.102, 95% CI 1.052 to 1.153; p=0.000) was independently associated with the presence of DCM before NT-proBNP was added in the multivariable regression model. CONCLUSIONS: The results indicate that chemerin is a novel biomarker of DCM.

Detection of a novel missense mutation in the mevalonate kinase gene in one Chinese family with DSAP.

Disseminated superficial actinic porokeratosis (DSAP) is the most common form of porokeratosis and a severe chronic autosomal dominant cutaneous disorder with high genetic heterogeneity. Recently, the mevalonate kinase (MVK) gene has been identified as a candidate gene responsible for DSAP and multiple mutations have been reported. Here, we report identification of a novel missense mutation in the MVK gene in a Chinese family with DSAP. A 50-year-old male was diagnosed as proband of DSAP based on the clinical and histological findings, which show numerous hyperpigmented macules by physical examination and cornoid lamella by skin biopsy. Similar skin symptoms were also observed in his father, who died many years ago. We prepared genomic DNA from the proband, unaffected individuals from his family members, as well as 100 unrelated healthy controls. PCR was then conducted using the above genomic DNA as template and the MVK gene-specific primers. The PCR product was subjected to direct sequencing and the sequence was compared to that of MVK gene within the NCBI database. We detected a heterozygous C to G transition at nucleotide 643 in exon 7 of MVK gene of the proband. This will result in an amino acid change at codon 215 (P.Arg215Gly.), which is from an arginine codon (CGA) to a Glycine codon (GGA). We did not detect any mutation in the unaffected family members or the 100 unrelated healthy controls, demonstrating that this is a novel missense mutation in MVK gene and therefore, contributes to the molecular diagnosis of DSAP.

Effects of aggregation and the surface properties of gold nanoparticles on cytotoxicity and cell growth.

The effect of gold nanoparticles (Au NPs) on cells remains open for investigation. Here we show that small Au NPs can be endocytosed by cells and form aggregates inside the cell, resulting in cytotoxicity. When the aggregates become too large to enter the cell and instead adhere onto the cell surface, however, the growth rate of HeLa cells increases. Printed patterns of Au NPs fabricated through inkjet printing technology were used to study the effects of Au NP aggregation on human cervical carcinoma (HeLa) cell activity. The growth of the HeLa cells was inhibited on the polymer-coated Au NPs but increased on the silicon substrate. On the uncoated Au NP surface, however, the HeLa cell growth rate was higher than that on the silicon substrate. Experiments with Escherichia coli cells showed a similar effect of the Au NPs. This phenomenon provides a new perspective for research on toxicity in nanoparticle biology. FROM THE CLINICAL EDITOR: Printed patterns of Au NPs fabricated through inkjet printing technology were used to study the effects of Au NP aggregation on human cervical carcinoma (HeLa) cell activity. Small Au NPs can be endocytosed by cells resulting in cytotoxicity; in contrast, large aggregates adhere onto the cell surface and increase the growth rate of HeLa cells.

A single-nucleotide polymorphism of the TNFSF4 gene is associated with systemic lupus erythematosus in Chinese Han population.

Systemic lupus erythematosus (SLE) is a prototypic autoimmune disease with complex genetic inheritance. Recently, single-nucleotide polymorphisms (SNPs) in tumor necrosis factor (TNF) superfamily gene TNFSF4 have been shown to be associated with SLE in European and Hong Kong Chinese populations. But it is unknown whether it is also associated with the disease in Mainland Chinese Han population. We genotyped the SNPs rs1234315 near the TNFSF4 gene in 1,344 SLE patients and 4,315 controls of Chinese Han population and confirmed the association between the SNP and the SLE [odds ratios (ORs) of 1.45 and P values of 1.5 × 10(-16)]. The stratification analyses showed that rs1234315 was more strongly associated with SLE patients with arthritis. Our study not only suggested that the TNFSF4 gene was associated with SLE in Chinese Han population, but also implied that it might be a common genetic factor predisposing to the development of SLE in multiple populations.

A single-nucleotide polymorphism of the TNFAIP3 gene is associated with systemic lupus erythematosus in Chinese Han population.

Systemic lupus erythematosus (SLE) is a complex systemic disease influenced by genetic and environmental factors. The exact pathogenesis of SLE is still unknown. Recently, several genome-wide association studies (GWA) in European population have found many novel susceptibility genes for SLE including TNFAIP3. In order to examine whether TNFAIP3 is associated with SLE in Chinese Han population, we genotyped one of its non-synonymous mutation SNP rs2230926, showing significant association evidence with SLE in European population, with 1,420 cases and 4,461 controls of Chinese Han by using Sequenom MassArray system. Highly significant association between SNP rs2230926 and SLE of Chinese Han was detected [OR = 1.65, 95% confidence interval (CI): 1.392-1.986, P = 2.03 x 10(-8)]. Interestingly, rs2230926 of TNFAIP3 was also associated with arthritis, ANA and some other subphenotypes of the disease. Our findings suggest that SNP rs2230926 in the TNFAIP3 might be a common genetic factor for SLE within different populations in terms of Chinese Han and European population.

Association of HLA class I alleles with keloids in Chinese Han individuals.

Keloids are benign fibroproliferative dermal tumors of unknown etiology. Some studies have suggested that human HLA status might potentiate development of keloids phenotype. No report has been published about HLA class I alleles associated with keloids in Chinese Han individuals. To investigate the etiology of keloids, the polymerase chain reaction-sequence-specific primer (PCR-SSP) method was used to analyze the distribution of HLA class I alleles in 192 patients with keloids and 252 healthy controls in a Chinese Han population. The frequencies of HLA-A*03 (6.77% vs 0%, p(c) < 10(-7)), A*25 (10.16% vs 4.56%, p(c) = 0.0111), B*07 (7.81% vs 2.58%, p(c) = 0.0080), and Cw*0802 (19.79% vs 10.32%, p(c) = 0.0004) were significantly increased in keloid patients, whereas the frequency of HLA-A*01 (18.75% vs 38.10%, p(c) < 10(-7)) was highly decreased, compared with that in healthy controls. The A*03-B*07, A*25-B*07, A*03-Cw*0802, A*25-Cw*0802, and B*07-Cw*0802 were found as high-risk haplotypes in developing keloids in this study. No extended haplotype was found to be significantly related to keloids. Through stratified analysis, the association of subgroups (single site/multiple site, severity, and family history) of keloid patients with specific HLA alleles was identified. Our data suggest these alleles may be keloids susceptibility genes or may be in close linkage with the susceptibility genes.

[Effects of allergen and intranasal glucocorticoid on Th17 and RORgamma t in peripheral blood in patients with allergic rhinitis.].

OBJECTIVE: To study the effect of specific immunotherapy and intranasal glucocorticoid on T help 17 (Th17) cells and RORgammat in peripheral blood in patients with allergic rhinitis (AR). METHODS: Forty patients with allergic rhinitis (group A) were divided randomly into two subgroups (group A1 and A2), and each subgroup had 20 patients. The patients in group A1 were treated with intranasal glucocorticoid (INGS) for one-year. The patients in group A2 were treated with special immunotherapy (SIT) for 4 weeks. Blood samples were respectively taken from 10 healthy individuals (group B), 20 AR patients (group A1) before and after SIT with specific standardized allergen and 20 AR patients (group A2) before and after INGS. The ratio of Th17 cells in peripheral blood monouclear cells (PBMC) were analysed by flow cytometry. The expression of RORgammat mRNA were detected by real-time polymerase chain reaction and the interleukin-23(IL-23), IL-17, IL-6 were detected by enzyme-linked immunosorbent assay. RESULTS: The ratio of Th17 cells in PBMC and the expression of RORgammat mRNA in group A [(18.97 +/- 1.05)% and (0.604 +/- 0.027)] were respectively higher than those in group B [(15.12 +/- 1.09)% and (0.447 +/- 0.024)] and the difference reached statistical significance (t were respectively -10.056 and -17.986, each P < 0.01). The level of IL-6, IL-17 and IL-23 in group A were respectively higher than those in group B and the difference reached statistical significance (t were respectively -41.149, -17.618 and -26.824, all P < 0.01). The ratio of Th17 cells in PBMC, the expression of RORgammat mRNA, the level of IL-6, IL-17 and IL-23 before INGS did not show significant difference from those of after INGS in group A1 (t were respectively 0.298, 0.240, -1.136, 0.283 and -1.670, all P > 0.05). The ratio of Th17 cells in PBMC and the expression of RORgammat mRNA were respectively (18.99 +/- 1.14)% and (0.603 +/- 0.027) before SIT and were respectively (16.30 +/- 1.63)% and (0.429 +/- 0.023) after SIT in group A2, and the difference reached statistical significance (t were respectively 6.035 and 22.015, all P < 0.01). The level of IL-6, IL-17 and IL-23 before SIT were lower respectively than those of after SIT in group A2 and the difference reached statistical significance (t were respectively 9.235, 11.289, 7.267, all P < 0.01). CONCLUSIONS: The ratio of Th17 cells in PBMC, the expression of RORgammat mRNA, the level of IL-6, IL-17 and IL-23 were up-regulated in patients with AR. The treatment of SIT could get the 5 items down and the treatment of INGS couldn't.

The role of G protein-coupled receptor 40 in lipoapoptosis in mouse beta-cell line NIT-1.

Free fatty acids (FFAs) exert divergent effects on beta-cells. Acute exposure to FFAs stimulates insulin secretion, whereas chronic exposure impairs beta-cell function and induces apoptosis. The G protein-coupled receptor 40 (GPR40) is preferentially expressed in beta-cells and is activated by a wide range of FFAs. In this study, we used small interfering RNA technology and apoptosis assay in mouse beta-cell NIT-1 to address the role of GPR40 in beta-cell lipoapoptosis and function. Results showed that palmitate induced beta-cell apoptosis, which was not mediated through GPR40, whereas oleate protected NIT-1 cells from palmitate-induced lipoapoptosis, which was mediated at least in part through GPR40. Moreover, by detecting the activation of the phosphatidylinositol 3-kinase and MAP kinase (MAPK) pathways, we found that oleate promoted the activation of extracellular signal-regulated protein kinase-MAPK pathway mainly via GPR40, increased the expression of early growth response gene-1, leading to the anti-lipoapoptotic effect on NIT-1 cells. It was suggested that GPR40 might be implicated in the control of beta-cell mass plasticity and GPR40 probably provide a link between obesity and type 2 diabetes.

[Genetical diagnosis in a family with X-linked hypohidrotic ectodermal dysplasia].

OBJECTIVE: To identify the mutations of ED1 gene in a family with X-linked hypohidrotic ectodermal dysplasia METHODS: Eight coding exons of ED1 gene of two patients with clinically confirmed X-linked hypohidrotic ectodermal dysplasia, their parents, and 100 unrelated population-matched control were amplified by polymerase chain reaction. The products were further analyzed by direct sequencing. RESULTS: Two patients with X-linked hypohidrotic ectodermal dysplasia in this pedigree showed a point mutation at nucleotide 1 045 ( A > G) . Meanwhile, heterozygous double peaks of nucleotide G and A at the same position were found in their mother, but not in their father and 100 unrelated population-matched controls. CONCLUSION: The c. 1 045A > G mutation of ED1 gene may be the pathologic cause of this Chinese family with X-linked hypohidrotic ectodermal dysplasia.

Estrogen receptor alpha gene polymorphism associated with type 2 diabetes mellitus and the serum lipid concentration in Chinese women in Guangzhou.

BACKGROUND: Estrogen might play an important role in type 2 diabetes mellitus pathogenesis. A number of polymorphisms have been reported in the estrogen receptor alpha (ERalpha) gene (also named ESR1), including the XbaI and PvuII restriction enzyme polymorphisms of ESR1, which may be involved in disease pathogenesis. The aim of this study was to determine whether ERX gene polymorphisms are associated with type 2 diabetes mellitus and serum lipid level. METHODS: Two hundred and ninety-nine patients with type 2 diabetes mellitus were compared with three hundred and forty-one health controls of Guangzhou in China, both were male and postmenopausal female residents at 51 - 70 years. ESR1 genotyping was performed using polymerase chain reaction (PCR) and PvuII and XbaI restriction fragment length polymorphism (PCR-RFLP) analysis. RESULTS: ESR1 allelic frequencies of P, p and X, x alleles were 0.408, 0.592; 0.360, 0.640 in the type 2 diabetes mellitus group and 0.318, 0.682; 0.328, 0.672 in the control group, respectively. In case-control study, there was significant difference in PvuII, but not XbaI, allele frequency between the type 2 diabetes mellitus and control groups (P = 0.001 and P = 0.122). When the group was separated into men and women, the difference was significant in women (P < 0.001) but not in men (P = 0.854) with the PvuII genotype, and the effect of PvuII variant on the development of type 2 diabetes mellitus was improved with aging. In addition, PvuII genotype was associated with blood glucose [fasting blood glucose (FBG), postprandial blood glucose (PBG)] and serum lipid [total cholesterol (TC) and low density lipoprotein (LDL)-c] concentration in healthy women. CONCLUSIONS: PvuII polymorphism of ESR1 increases susceptibility to type 2 diabetes mellitus in Chinese Guangzhou women. ESR1 variants may also impact serum lipid metabolism, which might provide a mechanism connecting ESR1 to type 2 diabetes.