Improving precision management of anxiety disorders: a Mendelian randomization study targeting specific gut microbiota and associated metabolites.

Background: There is growing evidence of associations between the gut microbiota and anxiety disorders, where changes in gut microbiotas may affect brain function and behavior via the microbiota-gut-brain axis. However, population-level studies offering a higher level of evidence for causality are lacking. Our aim was to investigate the specific gut microbiota and associated metabolites that are closely related to anxiety disorders to provide mechanistic insights and novel management perspectives for anxiety disorders. Method: This study used summary-level data from publicly available Genome-Wide Association Studies (GWAS) for 119 bacterial genera and the phenotype "All anxiety disorders" to reveal the causal effects of gut microbiota on anxiety disorders and identify specific bacterial genera associated with anxiety disorders. A two-sample, bidirectional Mendelian randomization (MR) design was deployed, followed by comprehensive sensitivity analyses to validate the robustness of results. We further conducted multivariable MR (MVMR) analysis to investigate the potential impact of neurotransmitter-associated metabolites, bacteria-associated dietary patterns, drug use or alcohol consumption, and lifestyle factors such as smoking and physical activity on the observed associations. Results: Bidirectional MR analysis identified three bacterial genera causally related to anxiety disorders: the genus Eubacterium nodatum group and genus Ruminococcaceae UCG011 were protective, while the genus Ruminococcaceae UCG011 was associated with an increased risk of anxiety disorders. Further MVMR suggested that a metabolite-dependent mechanism, primarily driven by tryptophan, tyrosine, phenylalanine, glycine and cortisol, which is consistent with previous research findings, probably played a significant role in mediating the effects of these bacterial genera to anxiety disorders. Furthermore, modifying dietary pattern such as salt, sugar and processed meat intake, and adjusting smoking state and physical activity levels, appears to be the effective approaches for targeting specific gut microbiota to manage anxiety disorders. Conclusion: Our findings offer potential avenues for developing precise and effective management approaches for anxiety disorders by targeting specific gut microbiota and associated metabolites.

Glioma exosomal microRNA-148a-3p promotes tumor angiogenesis through activating the EGFR/MAPK signaling pathway via inhibiting ERRFI1.

BACKGROUND: Glioma is the most frequent and lethal primary brain malignancy. Amounting evidence has highlighted the importance of exosomal microRNAs (miRNAs or miRs) in this malignancy. This study aimed to investigate the regulatory role of exosomal miR-148a-3p in glioma. METHODS: Bioinformatics analysis was firstly used to predict the target genes of miR-148a-3p. Exosomes were then extracted from normal human astrocytes and glioma cells. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was applied to determine the expression patterns of miR-148a-3p and ERBB receptor feedback inhibitor 1 (ERRFI1). Dual-luciferase reporter gene assay was applied to verify the direct binding between miR-148a-3p and ERRFI1. Cell counting kit-8 and tube formation assays were further conducted to assess the proliferation and angiogenic properties of human umbilical vein endothelial cells (HUVECs) in the co-culture system with exosomes. Lastly, glioma tumor models were established in BALB/c nude mice to study the role of exosomal miR-148a-3p in vivo. RESULTS: miR-148a-3p was highly expressed, while ERRFI1 was poorly expressed in glioma. miR-148a-3p was found to be enriched in glioma cells-derived exosomes and could be transferred to HUVECs via exosomes to promote their proliferation and angiogenesis. ERRFI1 was identified as a target gene of miR-148a-3p. In addition, miR-148a-3p activated the epidermal growth factor receptor (EGFR)/mitogen-activated protein kinase (MAPK) signaling pathway by inhibiting ERRFI1. In the co-culture system, our data demonstrated that glioma cells-derived exosomal miR-148a-3p down-regulated ERRFI1 and activated the EGFR/MAPK signaling pathway, so as to promote cell proliferation and angiogenesis. In vivo experimentation further demonstrated that this mechanism was responsible for the promotive role of exosomal miR-148a-3p in tumorigenesis and angiogenesis. CONCLUSION: Taken together, glioma-derived exosomal miR-148a-3p promoted tumor angiogenesis through activation of the EGFR/MAPK signaling pathway by ERRFI1 inhibition.

Different levels of circulating Th22 cell and its related molecules in Graves' disease and Hashimoto's thyroiditis.

PURPOSE: The aim of this study was to make a comparative analysis of the possible different expression of Th22 cells in two subtypes of autoimmune thyroid diseases (AITDs), i.e., Graves' disease (GD) and Hashimoto's thyroiditis (HT). METHODS: We recruited 61 AITDs patients (31 GD and 30 HT) and 22 controls. Serum level of IL-22 was measured by enzyme linked immunosorbent assay (ELISA). The proportion of Th22 cells in peripheral blood mononuclear cells (PBMCs) was analyzed by flow cytometry. The messenger RNA (mRNA) expressions of IL-22, its receptors (IL10R2, IL22R1) and key transcription factor (aryl hydrocarbon receptor, AHR) in PBMCs were assayed by quantitative reverse transcription polymerase chain reaction (qRT-PCR). Several cytokines of the cultured PBMCs were also measured under IL-22 stimulation. RESULTS: The proportion of Th22 cells, serum IL22 level and IL-22 mRNA expression were significantly higher in patients with GD than in healthy controls. Additionally, AHR increased in GD patients compared to healthy controls. However, the elevation of Th22 cells and their relative cytokines was not found in patients with HT. Consistent with specific mRNAs expression of cultured PBMCs, IL-4 increment in supernatant was much higher in GD group than in control group, while IFN-γ levels were decreased under IL-22 stimulation. CONCLUSION: Th22 cells may participate in the pathogenesis of AITDs as a proinflammatory factor, especially in GD, through expressing and secreting IL-22.

Polymorphisms of the TNFAIP3 region and Graves' disease.

Autoimmune thyroid disease (AITD) is a multifactorial organ-specific autoimmune disorder, and both genetic susceptibility and environmental factors are involved in its etiology. TNFAIP3 encodes the ubiquitin-modifying enzyme (A20), a key regulator of inflammatory signaling pathways. The aim of the present study was to evaluate the association between TNFAIP3 gene polymorphisms and AITD in Chinese Han population. Three single nucleotide polymorphisms (SNPs) in TNFAIP3 gene locus (rs598493, rs610604 and rs661561) were detected in a set of 667 patients with AITD and 301 controls in Han Chinese population using the Matrix Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometer (MALDI-TOF-MS) Platform. Compared with those of the controls, the frequencies of GG genotype of rs598493, the AA genotype of rs610604, the allele G and GG genotype of rs661561 were significantly increased in Graves' disease (GD) patients. However, the frequencies of AG genotype of rs598493 and AC genotype of rs610604 were significantly decreased in GD patients. The ATC haplotype (rs598493, rs661561 and rs610604) was associated with a decreased risk of GD. No significant differences in the three SNPs were observed between HT patients and controls. Our study shows a clear association between the polymorphisms of TNFAIP3 gene and GD, not HT, suggesting that TNFAIP3 gene is likely to be a genetic susceptibility factor to GD.

Bipolar transurethral resection of the prostate causes deeper coagulation depth and less bleeding than monopolar transurethral prostatectomy.

OBJECTIVE: To investigate the hemostatic capability of mono- and bipolar transurethral resection of the prostate by comparing the perioperative blood loss with the coagulation depth achieved with mono- and bipolar transurethral resection of the prostate. METHODS: A total of 136 patients with lower urinary tract symptoms associated with benign prostatic hyperplasia were randomized to undergo transurethral resection of the prostate using either a monopolar system (Karl Storz, Co., Tuttlingen, Germany) or a gyrus PlasmaKinetic bipolar system (Gyrus-ACMI Corporation, Maple Grove, MN). The operative time, resected tissue weight, decline in serum sodium and hemoglobin, postoperative bleeding, and the coagulation depth were compared. RESULTS: There were no statistically significant differences in operative time, resected tissue weight, and capsular perforation. The decline in hemoglobin and serum sodium was 1.15 ± 0.53 g/dL and 4.57 ± 0.71 mmol/L in monopolar transurethral resection of the prostate group, respectively, whereas they fell only 0.71 ± 0.42 g/dL and 2.02 ± 0.53 mmol/L in the bipolar transurethral resection of the prostate group, respectively (P <.001). The rate of postoperative bleeding was significantly higher in the monopolar transurethral resection of the prostate group (P = .027). The coagulation depths with mono- and bipolar transurethral resection of the prostate were 127.56 ± 27.76 and 148.48 ± 31.64 µm, respectively (P <.001). CONCLUSION: Our results demonstrate that bipolar transurethral resection of the prostate causes less intraoperative hemoglobin drop and postoperative bleeding than monopolar transurethral resection of the prostate, which may be associated with the deeper coagulation depth of bipolar transurethral resection of the prostate.

Bacteriuria after bipolar transurethral resection of the prostate: risk factors and correlation with leukocyturia.

OBJECTIVES: To analyze the risk factors of postoperative bacteriuria and the correlation with leukocyturia after bipolar transurethral resection of the prostate (TURP). METHODS: A total of 121 noncatheterized patients with sterile preoperative urine undergoing bipolar TURP for benign prostatic hyperplasia (BPH) were entered into the prospective study. All patients received antibiotic prophylaxis with ceftriaxone. Two urine specimens of each patient, one for urinalysis (urinary leukocyte count) and one for urine culture, were collected on removal of the catheter, 1 and 4 weeks after surgery. The risk factors of postoperative bacteriuria and correlation with leukocyturia were investigated. RESULTS: The incidence of bacteriuria after bipolar TURP was 18.2% (22/121). Multivariate analysis documented 3 independent risk factors of postoperative bacteriuria: operating time >60 minutes (P = .014), duration of catheterization >3 days(P = .001), and disconnection of the closed urine drainage system (P <10(-3)). The mean leukocyte counts in urine were 405.3, 389.5, and 113.8/µL on removal of the catheter, 1 and 4 weeks after surgery, respectively. Of 363 urine specimens, the mean concentration of leukocytes with and without bacteriuria were 323.9 and 297.6/µL, respectively (P >.05). There was no significant correlation between bacteriuria and leukocyturia (>10 leukocytes/high power field (P >.05). CONCLUSIONS: The results of our study have shown that the operating time, duration of catheterization, and disconnection of the closed urine drainage system may influence the occurrence of bacteriuria after bipolar TURP, and leukocyturia cannot reflect the possibility of postoperative bacteriuria.

[Bipolar transurethral resection of the prostate versus monopolar transurethral prostatectomy: a pathological study in a canine model].

OBJECTIVE: To compare the postoperative depths of the coagulation zones and pathological changes between bipolar transurethral resection of the prostate with plasmakinetic energy (PKRP) and monopolar transurethral prostatectomy (TURP) in canines. METHODS: Twenty-five male dogs were randomly divided into a PKRP group (n = 12), a TURP group (n = 12) and a sham-operation control group (n = 1). The dogs were sacrificed, their prostates harvested at 0 week (immediately after surgery), 1 week, 2 weeks and 8 weeks postoperatively and sectioned for pathologic analysis and measurement of the coagulation zones. RESULTS: At 0, 1 and 2 weeks after the operation, the coagulation depths were (237.73 +/- 20.12) microm, (113.03 +/- 16.65) microm and (106.01 +/- 16.36) microm in the PKRP group, and (200.75 +/-19.34) microm, (129.46 +/- 17.81) microm and (116.04 +/- 25.67) microm in the TURP group (P < 0.01). At 8 weeks, the coagulation zones completely peeled off and the wounds were covered by regenerated urothelial in both of the groups. At 0, 1, 2 and 8 weeks, different inflammatory reactions were observed in the prostates of the PKRP and TURP groups, with some glandular lumens beneath the coagulation zones expanded and epithelia damaged. However, none of these phenomena occurred in the sham-operation control group. CONCLUSION: Pathologically, PKRP and TURP inflicted basically similar effects on the prostate of the canine. However, the coagulation zone was deeper intraoperatively and became thinner postoperatively with the former than with the latter, which suggests that PKRP causes less bleeding and less penetrative thermal damage than TURP.

Association of the thyroglobulin gene polymorphism with autoimmune thyroid disease in Chinese population.

OBJECTIVE: This study was performed to identify the presence of previously reported thyroglobulin (Tg) gene single nucleotide polymorphisms (SNPs) in Han Chinese Asians, and to investigate their potential relation to autoimmune thyroid disease (AITD). METHODS: Polymorphisms were determined by polymerase chain reaction-restriction fragment length polymorphism analysis (PCR-RFLP) in 228 Chinese patients with AITD (146 with Graves' disease and 82 with Hashimoto's thyroiditis) and 131 healthy Chinese controls. RESULTS: (1) The occurrence of four common Tg gene SNPs (E10SNP24 T/G and E10SNP158 T/C in exon 10, E12SNP A/G in exon 12, and E33SNP C/T in exon 33) was confirmed in this Chinese population. No differences in allele and genotype frequencies were found between AITD patients and control subjects, or between male and female individuals in any group. Neither were differences in allele frequencies observed when Graves' disease (GD) or Hashimoto's thyroiditis (HT) patients were analyzed separately. (2) Haplotype analysis of these four SNPs revealed that the G-C-A-C haplotype was significantly associated with HT (P < 0.01, OR = 3.06, OR 95% CI [1.326-7.089]) and with serum anti-Tg antibody (Tg-Ab) positive AITD patients (P = 0.028, OR = 3.34). CONCLUSION: Our study confirms the existence of four SNPs among Han Chinese. In addition, the association of one SNP haplotype with HT suggests that Tg may be an AITD susceptibility gene.

[Association of polymorphism of protein tyrosine phosphatase nonreceptor-22 gene with AITD].

AIM: To evaluate the association of PTPN22 gene polymorphism with autoimmune thyroid disease (AITD) in Chinese people and to analyze the relationship between SNP of CTLA-4 gene and SNP of PTPN22 gene. METHODS: 149 patients with Graves' disease (GD) and 82 patients with Hashimoto's thyroiditis (HT) as well as 131 healthy people as controls were investigated. PTPN22 gene polymorphism +1858 C>T and CTLA-4 gene polymorphism 49A>G were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). PTPN22 gene polymorphism -1123G>C at promoter was genotyped by single allele-specific primer polymerase chain reaction (SASP-PCR). RESULTS: (1) +1858C>T for PTPN22 gene was not polymorphic enough in patients and controls. (2) Statistic differences in alleles and genotype frequency of -1123G>C were observed between GD patients and controls (P=0.040, 0.013; OR=1.44, 2.33, respectively). (3) Differences in alleles and genotype frequency of 49A>G for CTLA-4 gene were observed in patients and controls. (4) Individuals with PTPN22 CC genotype and CTILA-4 G alleles had an increased risk of developing GD (OR=3.31, 95%CI: 2.69-8.89) compared with those with PTPN22 G alleles and CTLA-4 AA genotype. CONCLUSION: -1123 G>C SNP of PTPN22 gene is associated with GD. There is coordination between PTPN22 CC genotype and CTLA-4 G alleles in the development of GD.

[Effects of experimental varicocele on the apoptosis of epididymis epithelium and synthesizing function of the epididymis in rats].

OBJECTIVE: To study the relationships of experimental varicocele to the apoptosis of epididymis epithelium and changes of the contents of alpha-1,4-glucosidase and sialic acid from the unilateral epididymis in adolescent rats, and to investigate the effects of varicocele on the unilateral epididymis epithelium. METHODS: Experimental left varicocele models of 16 adult male Sprague-Dawley rats were obtained by partial ligation of the left renal vein. The epididymides were collected for detecting the apoptosis of epididymis epithelium and the contents of alpha-1,4-glucosidase and sialic acid by using spectrophotometry. RESULTS: Seven days after the establishment of the left varicocele model, the index of the apoptosis of the left epididymis epithelium was significantly higher (P < 0.001) and the contents of alpha-1,4-glucosidase and sialic acid significantly lower (P < 0.05, P < 0.005) in the experimental group than in the control. CONCLUSION: The results suggest that unilateral varicocele may increase the apoptosis of epididymis epithelium and the contents of alpha-1,4-glucosidase and sialic acid and subsequently affect the synthesizing and secretory function of the epididymis.

[Changes of nuclear factor-kappa gene binding expression in and apoptosis of spermatogenic epithelial cells in the restored testis after torsion: experiment with rats].

OBJECTIVE: To investigate the changes of nuclear factor-kappa gene binding (NF-kappaB) expression in and apoptosis of spermatogenic epithelial cells in the restored testis after torsion and analyze the relationship between them. METHODS: Sixteen male SD rats underwent torsion of the left testis clockwise at an angle of 720 degrees for 2 hours and then the testis was restored to the original position and fixed. Then the 16 rats were randomly divided into 2 equal group: Group I in which salicylazosulfapyridium (SASP) suspension was infused intra-gastrically 5 h after operation and then once a day for 4 times, and Group II in which normal saline (NS) was infused in the same manner. Eight rats (Group III) underwent sham operation and then infused with NS in the same manner as that of Group II. Three days after operation the rats were killed and the samples of the testes at the torsion side were taken out and the seminiferous tubules were isolated. Western blotting was used to detect the NF-kappaB expression in the cytoplasm and nucleus of spermatogenic epithelial cells. Immunohistochemistry was used to detect the in situ expression of NF-kappaB. The apoptosis of the spermatogenic epithelial cells was examined by TUNEL method. RESULTS: Western blotting showed that the NF-kappaB expression in the cytoplasm of spermatogenic epithelial cells of Group II was 9.4 +/- 2.68, somewhat lower, but not significantly, than those of Group I and III (12 +/- 2.2 and 11.1 +/- 3 respectively, both P > 0.05). The NF-kappaB expression in the nucleus of spermatogenic epithelial cells of Group II was 21.1 +/- 3.6, significantly higher than those of Group I and III (8.4 +/- 3.1 and 6.0 +/- 2.3 respectively, both P < 0.05). However, there were no significant differences in the NF-kappaB expression in the cytoplasm and nucleus of spermatogenic epithelial cells between Groups I and III. The NF-kappaB activity coefficient of spermatogenic epithelial cells of Group II was 2.32 +/- 0.4, significantly higher than those of Groups I and III (0.68 +/- 0.3 and 0.52 +/- 0.1 respectively, both P < 0.01). However, there was no significant difference in the NF-kappaB activity coefficient of spermatogenic epithelial cells between Groups I and III (P > 0.05). The NF-kappaB positive cell rate of Group II was 66.1% +/- 3.8%, significantly higher than those of Groups I and III (15.6% +/- 2.6% and 10.8% +/- 2.7%, both P < 0.01). The apoptotic cell rate of Group II was 37.2% +/- 3.3%, significantly higher than those of Groups I and III (7.7% +/- 2.0% and 5.9% +/- 1.7%, both P < 0.01). CONCLUSION: After the torsion of testis, NF-kappaB was activated and released from the nucleus into the cytoplasm, thus initiating the apoptosis of spermatogenic epithelial cells.

Sulfasalazine prevents apoptosis in spermatogenic cells after experimental testicular torsion/detorsion.

AIM: To determine whether sulfasalazine can prevent apoptosis in spermatogenic cells by preventing the activation of NF-kappaB in spermatogenic epithelium in experimental testicular torsion. METHODS: Thirty-two adult male Sprague-Dawley rats were subjected to unilateral 720 degree testicular torsion for durations of 0 h and 2 h, then the torsion was relieved. The ischemic/reperfused testes were collected for the detection of NF-kappaB expression with Western blotting and immunohistochemistry techniques, and detection of apoptosis with TUNEL techniques. RESULTS: The NF-kappaB coefficient of spermatogenic epithelium and the apoptosis index of spermatogenic cells were significantly different in the operation and the sham-operation groups after experimental testicular torsion (P<0.01). CONCLUSION: NF-kappaB activation of spermatogenic epithelium is related to apoptosis of spermatogenic cells. Sulfasalazine can prevent apoptosis in spermatogenic cells after the experimental testicular torsion through prevention of NF-kappaB activation.