Integrated Proteomics and Metabolomics Analysis in Pregnant Rat Hippocampus After Circadian Rhythm Inversion.

To investigate the changes in proteins, metabolites, and related mechanisms in the hypothalamus of pregnant rats after circadian rhythm inversion during the whole pregnancy cycle. A total of 12 Wistar female rats aged 7 weeks were randomly divided into control (six rats) and experimental (six rats) groups at the beginning of pregnancy. The control group followed a 12-h light and dark cycle (6 a.m. to 6 p.m. light, 6 p.m. to 6 a.m. dark the next day), and the experimental group followed a completely inverted circadian rhythm (6 p.m. to 6 a.m. light the next day, 6 a.m. to 6 p.m. dark). Postpartum data were collected until 7-24 h after delivery, and hypothalamus samples were collected in two groups for quantitative proteomic and metabolism analyses. The differential proteins and metabolites of the two groups were screened by univariate combined with multivariate statistical analyses, and the differential proteins and metabolites enriched pathways were annotated with relevant databases to analyze the potential mechanisms after circadian rhythm inversion. A comparison of postpartum data showed that circadian rhythm inversion can affect the number of offspring and the average weight of offspring in pregnant rats. Compared with the control group, the expression of 20 proteins and 37 metabolites was significantly changed in the experimental group. The integrated analysis between proteins and metabolites found that RGD1562758 and lysophosphatidylcholine acyltransferase 1 (LPCAT1) proteins were closely associated with carbon metabolism (choline, NAD+, L-glutamine, theobromine, D-fructose, and pyruvate) and glycerophospholipid metabolism (choline, NAD+, L-glutamine, phosphatidylcholine, theobromine, D-fructose, pyruvate, and arachidonate). Moreover, the Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that the differential metabolites enriched in adenosine triphosphate (ATP)-binding cassette (ABC) transporters. Our study suggested that circadian rhythm inversion in pregnant rats may affect the numbers, the average weight of offspring, and the expressions of proteins and metabolism in the hypothalamus, which may provide a comprehensive overview of the molecular profile of circadian rhythm inversion in pregnant groups.

Reduced Cell-Free Mitochondrial DNA Levels Were Induced by Antipsychotics Treatment in First-Episode Patients With Schizophrenia.

Cell-free mitochondrial DNA (cf-mtDNA) is a damage-associated molecular pattern that boosts the release of cytokines and induces the immune response of the body; therefore, it is closely related to mental diseases. This study aims to evaluate a potential link between cf-mtDNA and clinical progression in first-episode patients with schizophrenia. In this study, plasma cf-mtDNA levels in 34 first-episode patients with schizophrenia before and after 8 weeks of antipsychotic treatment were examined. In addition, the clinical progression of first-episode schizophrenia was assessed using the Positive and Negative Syndrome Scale (PANSS). The copy number changes in the plasma cf-mtDNA (Δcf-mtDNA) were significantly correlated with changes in the PANSS scale scores (ΔPANSS) in first-episode patients with schizophrenia (ΔPANSS total score, P = 0.002; ΔPANSS positive score, P = 0.01). Plasma cf-mtDNA may represent a relevant tool in the future to assist in the assessment of clinical progression in first-episode patients with schizophrenia.

Therapeutic effect of curcumin on experimental colitis mediated by inhibiting CD8+CD11c+ cells.

AIM: To verify whether curcumin (Cur) can treat inflammatory bowel disease by regulating CD8+CD11c+ cells. METHODS: We evaluated the suppressive effect of Cur on CD8+CD11c+ cells in spleen and Peyer's patches (PPs) in colitis induced by trinitrobenzene sulfonic acid. Mice with colitis were treated by 200 mg/kg Cur for 7 d. On day 8, the therapeutic effect of Cur was evaluated by visual assessment and histological examination, while co-stimulatory molecules of CD8+CD11c+ cells in the spleen and PPs were measured by flow cytometry. The levels of interleukin (IL)-10, interferon (IFN)-γ and transforming growth factor (TGF)-ß1 in spleen and colonic mucosa were determined by ELISA. RESULTS: The disease activity index, colon weight, weight index of colon and histological score of experimental colitis were obviously decreased after Cur treatment, while the body weight and colon length recovered. After treatment with Cur, CD8+CD11c+ cells were decreased in the spleen and PPs, and the expression of major histocompatibility complex II, CD205, CD40, CD40L and intercellular adhesion molecule-1 was inhibited. IL-10, IFN-γ and TGF-ß1 levels were increased compared with those in mice with untreated colitis. CONCLUSION: Cur can effectively treat experimental colitis, which is realized by inhibiting CD8+CD11c+ cells.

Curcumin Suppressed Activation of Dendritic Cells via JAK/STAT/SOCS Signal in Mice with Experimental Colitis.

Dendritic cells (DCs) play a pivotal role as initiators in the pathogenesis of inflammatory bowel disease and are regulated by the JAK/STAT/SOCS signaling pathway. As a potent anti-inflammatory compound, curcumin represents a viable treatment alternative or adjunctive therapy in the management of chronic inflammatory bowel disease (IBD). The mechanism of curcumin treated IBD on DCs is not completely understood. In the present study, we explored the mechanism of curcumin treated experimental colitis by observing activation of DCs via JAK/STAT/SOCS signaling pathway in colitis mice. Experimental colitis was induced by 2, 4, 6-trinitrobenzene sulfonic acid. After 7 days treatment with curcumin, its therapeutic effect was verified by decreased colonic weight, histological scores, and remitting pathological injury. Meanwhile, the levels of major histocompatibility complex class II and DC costimulatory molecules (CD83, CD28, B7-DC, CD40, CD40 L, and TLR2) were inhibited and followed the up-regulated levels of IL-4, IL-10, and IFN-γ, and down-regulated GM-CSF, IL-12p70, IL-15, IL-23, and TGF-ß1. A key finding was that the phosphorylation of the three members (JAK2, STAT3, and STAT6) of the JAK/STAT/SOCS signaling pathway was inhibited, and the three downstream proteins (SOCS1, SOCS3, and PIAS3) from this pathway were highly expressed. In conclusion, curcumin suppressed the activation of DCs by modulating the JAK/STAT/SOCS signaling pathway to restore immunologic balance to effectively treat experimental colitis.

Curcumin improves regulatory T cells in gut-associated lymphoid tissue of colitis mice.

AIM: To explore the probable pathway by which curcumin (Cur) regulates the function of Treg cells by observing the expression of costimulatory molecules of dendritic cells (DCs). METHODS: Experimental colitis was induced by administering 2, 4, 6-trinitrobenzene sulfonic acid (TNBS)/ethanol solution. Forty male C57BL/6 mice were randomly divided into four groups: normal, TNBS + Cur, TNBS + mesalazine (Mes) and TNBS groups. The mice in the TNBS + Cur and TNBS +Mes groups were treated with Cur and Mes, respectively, while those in the TNBS group were treated with physiological saline for 7 d. After treatment, the curative effect of Cur was evaluated by colonic weight, colonic length, weight index of the colon, and histological observation and score. The levels of CD4(+)CD25+Foxp3(+) T cells (Treg cells) and costimulatory molecules of DCs were measured by flow cytometry. Also, related cytokines were analyzed by enzyme-linked immunosorbent assay. RESULTS: Cur alleviated inflammatory injury of the colonic mucosa, decreased colonic weigh and histological score, and restored colonic length. The number of Treg cells was increased, while the secretion of TNF-α, IL-2, IL-6, IL-12 p40, IL-17 and IL-21 and the expression of costimulatory molecules (CD205, CD54 [ICAM-1], TLR4, CD252[OX40 L], CD256 [RANK] and CD254 [RANK L]) of DCs were notably inhibited in colitis mice treated with Cur. CONCLUSION: Cur potentially modulates activation of DCs to enhance the suppressive functions of Treg cells and promote the recovery of damaged colonic mucosa in inflammatory bowel disease.

Astragalus polysaccharide attenuates rat experimental colitis by inducing regulatory T cells in intestinal Peyer's patches.

AIM: To explore probable mechanism underlying the therapeutic effect of Astragalus polysaccharide (APS) against experimental colitis. METHODS: Thirty-two Sprague-Dawley rats were randomly divided into four groups. Colitis was induced with 2, 4, 6-trinitrobenzene sulfonic acid (TNBS). The rats with colitis were treated with 400 mg/kg of APS for 7 d. The therapeutic effect was evaluated by colonic weight, weight index of the colon, colonic length, and macroscopic and histological scores. The levels of regulatory T (Treg) cells in Peyer's patches were measured by flow cytometry, and cytokines in colonic tissue homogenates were analyzed using enzyme-linked immunosorbent assay. The expression of related orphan receptor-γt (ROR-γt), IL-23 and STAT-5a was measured by Western blot. RESULTS: After 7-d treatment with APS, the weight index of the colon, colonic weight, macroscopical and histological scores were decreased, while the colonic length was increased compared with the model group. The expression of interleukin (IL)-2, IL-6, IL-17, IL-23 and ROR-γt in the colonic tissues was down-regulated, but Treg cells in Peyer's patches, TGF-ß and STAT5a in the colonic tissues were up-regulated. CONCLUSION: APS effectively ameliorates TNBS-induced experimental colitis in rats, probably through restoring the number of Treg cells, and inhibiting IL-17 levels in Peyer's patches.

[Regulatory effect of curcumin on subsets and co-stimulatory molecules of dendritic cells in spleen from mice with colitis].

OBJECTIVE: To explore the effect of curcumin (Cur) on the subpopulation and costimulatory molecules of dendritic cells (DCs) in spleen from colitis mice. METHODS: Forty male C57/BL mice were randomly divided into 4 groups: normal group, model group, Cur group and mesalazine group. Colitis was induced by 2, 4, 6-trinitrobenzene sulfonic acid (TNBS). On the 8th day following 7-day treatment with Cur, the mice were sacrificed and the length and mass of mouse colon were measured. And then the spleens were separated to prepare DCs. The numbers of CD205(+), CD4(+)CD205(+), CD8(+)CD205(+) DCs and the levels of major histocompability complex II (MHC-II), Toll-like receptor 2 (TLR2), TLR4 and CD83 expression were detected by flow cytometry. And we observed the pathological injury of colon. RESULTS: Compared with model group, the colonic mass index was reduced, the length of colon was lengthened, and the pathological injury was remitted. Meanwhile, the percents of CD205(+), CD4(+)CD205(+), CD8(+)CD205(+) DCs and the levels of MHC-II, TLR2 and TLR4 were down-regulated, however, the CD83 expression was up-regulated. CONCLUSION: Treatment efficacy of Cur on experimental colitis might be involved in regulating subpopulation and co-stimulatory molecules of dendritic cells in spleen from mice with colitis.

Si Shen Wan Regulates Phospholipase Cγ-1 and PI3K/Akt Signal in Colonic Mucosa from Rats with Colitis.

The present study explored the feasible pathway of Si Shen Wan (SSW) in inhibiting apoptosis of intestinal epithelial cells (IECs) by observing activation of phospholipase Cγ-1 (PLC-γ1) and PI3K/Akt signal in colonic mucosa from rats with colitis. Experimental colitis was induced by 2,4,6-trinitrobenzene sulfonic acid (TNBS) in the Sprague-Dawley rats. After SSW was administrated for 7 days after TNBS infusion, western blot showed an increment in levels of PI3K, p-Akt, and IL-23 and a decrement in levels of PLC-γ1 and HSP70 in colonic mucosal injury induced by TNBS. Meanwhile, assessments by ELISA revealed an increment in concentrations of IL-2, IL-6, and IL-17 and a reduction in level of TGF-ß after TNBS challenge. Impressively, treatment with SSW for 7 days significantly attenuated the expressions of PI3K and p-Akt and the secretion of IL-2, IL-6, IL-17, and IL-23 and promoted the activation of PLC-γ1, HSP70, and TGF-ß. Our previous studies had demonstrated that SSW restored colonic mucosal ulcers by inhibiting apoptosis of IECs. The present study demonstrated that the effect of SSW on inhibiting apoptosis of IECs was realized probably by activation of PLC-γ1 and suppression of PI3K/Akt signal pathway.

[A comparative study of Bolton index in the youth and middle-aged and elderly people].

PURPOSE: To Investigate the changes of Bolton index, maxillary and mandibular teeth size and teeth size discrepancy (TSD) in the youth and middle- aged and elderly people. METHODS: One hundred ninety-one youth dental plaster casts from malocclusion patients and 63 middle-aged and elderly people with snoring or obstructive sleep apnea-hypopnea syndrome (OSAHS) patients were selected. 12 teeth size between the upper and lower first molar, width of maxillary lateral incisor, Bolton anterior and overall ratios and teeth size discrepancy were measured and calculated. Statistical analysis was performed using SPSS 13.0 software package. RESULTS: There was no significant difference between the 2 groups in Bolton anterior and overall ratios (P<0.05 ).The mean size of the upper lateral tooth in the youth was larger than the middle-aged and elderly people(P<0.01).There was no significant difference in 6 upper or lower anterior teeth size between canine and 12 mandibular teeth size between first molar in 2 groups, but the mean teeth size in the middle-aged and elderly people was smaller.The prevalence of anterior and overall TSD in the youth was 31.4% and 42.4%,while it was 19.05% and 15.9% in the middle-aged and elderly group. There was significant difference(P<0.05) between 2 groups in width of upper lateral incisor, which was 7.46mm in the youth and 7.20 mm in the middle-aged and elderly group. Prevalence of laminal width of upper lateral incisor in patients with teeth size discrepancy was low. CONCLUSIONS: Bolton index does not change by age. Although the teeth size in middle-aged and elderly group are smaller than the youth group, the total reduction is little without clinical significance. The laminal width of upper lateral incisor isn't the main cause for anterior or overall dental size discrepancy. Supported by Research Fund from Department of Public Health of Guangxi Zhuang Autonomous Region (GWZ2013405 and GWZ2007149).

Si Shen Wan Inhibits mRNA Expression of Apoptosis-Related Molecules in p38 MAPK Signal Pathway in Mice with Colitis.

Si Shen Wan (SSW) is used to effectively treat ulcerative colitis (UC) as a formula of traditional Chinese medicine. To explore the mechanism of SSW-inhibited apoptosis of colonic epithelial cell, the study observed mRNA expression of apoptosis-related molecules in p38 MAPK signal pathway in colonic mucosa in colitis mice treated with SSW. Experimental colitis was induced by 2,4,6-trinitrobenzene sulfonic acid (TNBS) in mice; meanwhile, the mice were administrated daily either SSW (5 g/kg) or p38 MAPK inhibitor (2 mg/kg) or vehicle (physiological saline) for 10 days. While microscopical evaluation was observed, apoptosis rate of colonic epithelial cell and mRNA expression of apoptosis-related molecules were tested. Compared with colitis mice without treatment, SSW alleviated colonic mucosal injuries and decreased apoptosis rate of colonic epithelial cell, while the mRNA expressions of p38 MAPK, p53, caspase-3, c-jun, c-fos, Bax, and TNF- α were decreased in the colonic mucosa in colitis mice treated with SSW, and Bcl-2 mRNA and the ratio of Bcl-2/Bax were increased. The present study demonstrated that SSW inhibited mRNA expression of apoptosis-related molecules in p38 MAPK signal pathway to downregulate colonic epithelial cells apoptosis in colonic mucosa in mice with colitis.

Impact of catechol-o-methyltransferase polymorphisms on risperidone treatment for schizophrenia and its potential clinical significance.

OBJECTIVES: The main aim was to study the effects of COMT polymorphisms on response of risperidone treatment for schizophrenia and investigate the correlation between memory function of schizophrenia patients and COMT polymorphisms. DESIGN AND METHODS: Subjects were 83 schizophrenic patients who were antipsychotic drug free at the initiation of this study. Peripheral blood samples were obtained to identify COMT polymorphisms by using a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. Clinical Global Impressions (CGI), Positive and Negative Syndrome Scale (PANSS) and Wechsler Memory Scale (WMS) test were used to assess the effect of risperidone treatment. RESULTS: The Val/Met carriers showed a significant increase in change of P300 during treatment (P=0.032). Association of Val/Met carriers performed better than other genotypes (P=0.028). The mean plasma concentration of prolactin of Val/Val carriers was significantly lower (P=0.017). CONCLUSIONS: The COMT polymorphisms may be a potential biomarker for clinical risperidone treatment in schizophrenia.

[Treatment of skeletal Class II adult patients with microscrew implant anchorage and multi-loop edgewise arch wire].

OBJECTIVE: To evaluate the effects and mechanisms of the microscrew implant anchorage (MIA) combined with multi-loop edgewise arch wire (MEAW) technique in the treatment of skeletal Class II adult patients. METHODS: Eleven adult patients with skeletal Class II high-angle malocclusions were treated with fixed appliances. The spaces were closed by the springs from the MIA to the hook on the archwire. The height of the hook and the direction of the force were different according to the intrusion and retraction of upper anterior teeth. In the finishing stage, MEAW technique and modified class II elastics (from the first loop of MEAW to the MIA) were used for final detailing. Cephalometric analysis was used to evaluate the effect after treatment. RESULTS: After treatment, the decrease of SNA, ANB and FMA were (2.86 +/- 1.05) degrees , (2.82 +/- 0.96) degrees and (2.95 +/- 1.35) degrees , respectively. The torque control of upper anterior teeth was good. The protrusion of lower incisors and the molar extrusion were avoided. The upper molars were moved distally by (3.00 +/- 2.19) mm. CONCLUSIONS: The treatment of adult patients with skeletal Class II high angle malocclusions with MIA and MEAW technique could not only improve the facial esthetics but also avoided the common side effects of traditional Class II elastics.

[A study of Bolton tooth-size discrepancies of malocclusion patients].

OBJECTIVE: To analysis the sum and frequencies of Bolton tooth-size discrepancies Angle Class I, II, III malocclusion patients. METHODS: Measured each tooth crown mesial-distal size between the first molar of 439 dental plaster casts of malocclusion patients and obtained the sums of six anterior tooth-size and twelve tooth-size between the first molar of upper or lower and calculated the anterior and total tooth-size discrepancies by Bolton rate standards, then statistic analyses were done. RESULTS: Patients whose sums of anterior tooth-size discrepancy were pass 1.5 mm or less than -1.5 mm was 14.02% in Angle Class I malocclusion patients and 9.49% in Class II and 19.32% in Class III. Patients whose sums of total tooth-size discrepancy were pass 1.5 mm or less than -1.5 mm was 19.63% in Angle Class I malocclusion patients and 15.33% in Class II and 20.45% in Class III. The upper sum of anterior tooth-size of Class I malocclusion patients whose sums of anterior tooth-size discrepancies were pass 1.5 mm or less than -1.5 mm was always smaller than normal and the lower sum was always larger. The sums of anterior tooth-size discrepancy of Class I, II, III and total tooth-size discrepancy of Class I were always between 1.5 mm-2.5 mm or -2.5 mm(-)-1.5 mm. Patients whose sums of anterior tooth-size discrepancy were pass 3.5 mm or less than -3.5 mm was 2.34% in Class I malocclusion patients and 0 in class II and 0 in class III. Patients whose sums of total tooth-size discrepancy were pass 3.5 mm or less than -3.5 mm was 4.21% in Class I and 1.46% in Class II and 4.54% in Class III. CONCLUSION: Tooth-size discrepancy of malocclusion patients was not the general cause of malocclusion.