Microglial SIX2 suppresses lipopolysaccharide (LPS)-induced neuroinflammation by up-regulating FXYD2 expression.

Parkinson's disease (PD) is a severe neurodegenerative disease associated with the loss of dopaminergic (DA) neurons in the substantia nigra (SN). Although its pathogenesis remains unclear, microglia-mediated neuroinflammation significantly contributes to the development of PD. Here we showed that the sine oculis homeobox (SIX) homologue family transcription factors SIX2 exerted significant effects on neuroinflammation. The SIX2 protein, which is silenced during development, was reactivated in lipopolysaccharide (LPS)-treated microglia. The reactivated SIX2 in microglia mitigated the LPS induced inflammatory effects, and then reduced the toxic effect of conditioned media (CM) of microglia on co-cultured MES23.5 DA cells. Using the LPS-stimulated Cx3cr1-CreERT2 mouse model, we also demonstrated that the highly-expressed SIX2 in microglia obviously attenuated neuroinflammation and protected the DA neurons in SN. Further RNA-Seq analysis on the inflammatory activated microglia revealed that the SIX2 exerted these effects via up-regulating the FXYD domain containing ion transport regulator 2 (FXYD2). Taken together, our study demonstrated that SIX2 was an endogenous anti-inflammatory factor in microglia, and it exerted anti-neuroinflammatory effects by regulating the expression of FXYD2, which provides new ideas for anti-neuroinflammation in PD.

Dephosphorylation of Six2Y129 protects tyrosine hydroxylase-positive cells in SNpc by regulating TEA domain 1 expression.

Parkinson's disease (PD) is a neurodegenerative disease characterized by selective loss of dopaminergic (DA) neurons in the substantia nigra pars compacta (SNpc). We recently reported that Six2 could reverse the degeneration of DA neurons in a dephosphorylation state. Here we further identified that Eya1 was the phosphatase of Six2 that could dephosphorylate the tyrosine 129 (Y129) site by forming a complex with Six2 in damaged DA cells. Dephosphorylated Six2 then translocates from the cytoplasm to the nucleus. Using ChIP-qPCR and dual luciferase assay, we found that dephosphorylated Six2 down-regulates TEA domain1 (Tead1) expression, thus inhibiting 6-hydroxydopamine (6-OHDA)-induced apoptosis in DA cells. Furthermore, we showed Six2Y129F/Tead1 signaling could protect against the loss of SNpc tyrosine hydroxylase-positive (TH+) cells and improve motor function in PD model rats. Our results demonstrate a dephosphorylation-dependent mechanism of Six2 that restores the degeneration of DA neurons, which could represent a potential therapeutic target for PD.

Effect of plant-self debris on nitrogen removal, transformation and microbial community in mesocosm constructed wetlands planted with Myriophyllum aquaticum.

Aquatic macrophytes debris decomposition could release pollutants and nutrients into the water of constructed wetlands (CWs), but their role in nitrogen removal and transformation remains poorly understood. The present study investigated the effects of plant-self debris on nitrogen removal and microbial communities in mesocosm CWs planted with Myriophyllum aquaticum. During the 68-day operation, the plant debris addition did not change the mean removal efficiency of ammonium (NH4+-N) and total nitrogen (TN) of CWs but showed significant differences over the operation time. The NH4+-N and organic nitrogen released from the plant debris decomposition affected the nitrogen removal. The plant debris decreased the effluent nitrate concentration and N2O emission fluxes of the CWs with the increased abundance of denitrifying bacterial genera, indicating that plant debris decomposition increased the denitrification activities via dissolved organic carbon release. High-throughput sequencing indicated that the plant debris altered the distribution and composition of the microbial community in the sediments. Proteobacteria was the dominant phylum (28-52%), and the relative abundance of denitrifying bacteria genera was significantly higher in the sediments with debris addition (37-40%) than in the non-addition (6.6-7.7%). The present study provided new insights into the role of macrophytes in pollutant removal and the plant management strategy of CWs.

Identification of hepatitis B virus aetiologic antigens, HBx and Pre-S2, in diffuse large B-cell lymphoma.

Diffuse large B-cell lymphoma (DLBCL) has been reported to have a significant association with the hepatitis B virus (HBV) infection. However, there has been no experimental evidence to determine whether the components of the hepatitis B virus are expressed in lymphoid cells. In this study, we used immunohistochemical methods to explore whether the antigens of hepatitis B virus are expressed in DLBCL lymphoma cells in HBsAg-positive DLBCL patients (HBsAg + DLBCL). HBx antigen was detected in 48.9% of HBsAg + DLBCL patients, and the expression rate of the Pre-S2 antigen was 57.2%. HBx expression was significantly associated with high-level expression of c-Myc, while the Pre-S2 antigen was not. In this study, we demonstrated that HBx antigen and Pre-S2 antigen could be detected in lymphoma cells, and HBx expression was related to c-Myc expression. Our findings provide a strong basis for further study of the HBV-infected DLBCL and molecular mechanism underlying the lymphomagenesis.

Nitrogen removal performance and needed area estimation of surface-flow constructed wetlands using a probabilistic approach.

Pollutant concentrations in influents into constructed wetlands (CWs) are highly fluctuating and may vary over several orders of magnitude, leading to large uncertainties in removal performance assessment when using pollutant concentrations in the influent and effluent directly. Incorporating a probabilistic approach into removal performance assessment and needed area estimation of CWs could advantage decision making regarding wastewater treatment and engineering applications. A series of three-stage surface-flow CWs (SFCWs) were constructed for treating ammonium-rich swine wastewater. The surface removal rate and removal efficiency of ammonium nitrogen in the SFCWs using the probabilistic approach were 0.27-3.23 g m-2 d-1 and 43.0-99.9% (95% confidence interval (CI)), which were consistent with the deterministic approach (95% CI: 0.24-3.18 g m-2 d-1 and 70.4-99.9%). The needed SFCW area was estimated as 6.6 (95% CI: 1.4-17.8) to 29.7 (95% CI: 6.4-80.1) m2 for required removal efficiency from 40% to 90% for 0.18 m3 d-1 swine wastewater with different strengthens. For specific removal efficiency of 90%, the needed CW areas was 13.9 (95%CI: 4.9-42.7), 25.1 (95%CI: 5.9-66.0), 33.5 (95%CI: 13.5-87.1), and 40.8 (95%CI: 16.2-89.4) m2 for influent ammonium loading rate of 0.18-2.7, 2.7-14.4, 14.4-36, and 36-60 g d-1, respectively. The first-order removal constant of ammonium nitrogen decreased logarithmically with increasing influent and effluent concentration/loading rate in the SFCW units (p < 0.001), which was responsible for the needed SFCW areas covering a wide range. The reliability analysis confirmed the results from the probabilistic approach were appropriate. The present study shed new lights on the performance evaluation and design of CWs for treating wastewater with highly-fluctuating concentrations using a probabilistic approach.

Efficacy and safety of external application of Traditional Chinese Medicine for the treatment of acute gouty arthritis: a systematic review and Meta-analysis.

OBJECTIVE: To evaluate the clinical efficacy and safety of external application of Traditional Chinese Medicine (EATCM) on acute gouty arthritis (AGA). METHODS: Six electronic data bases were retrieved from their inception to march 2017. 15 randomized controlled clinical trials (RCTs) were included. The quality of the literatures was assessed according to Cochrane review criteria by using RevMan5.3 software; and related data was counted using Stata14.0 software. RESULTS: There was no significant difference in C-reactiveprotein (CRP) between experimental intervention group and control group [(WMD = 1.093, 95% CI (－2.514, 4.701); P = 0.553)]. In terms of overall efficacy, the EATCM's treatment group was significantly superior to control group [(OR = 3.692; 95% CI (2.408, 5.661); P < 0.001)]. The EATCM was better than western medicine treatments or other interventions in reducing the adverse reactions [(OR = 0.135; 95% CI (0.067, 0.274); P < 0.001)]. All these funnel plots showed unlikelihood of publishing bias. But due to excessive heterogeneity, the statistical results of serum uric acid and visual analogue scale and CRP between the two groups became uncertain. CONCLUSION: The results indicate that EATCM may have greater overall efficacy with fewer adverse drug reactions, although the evidence is weak owing to the low methodological quality and the small number of the included trials.

Replication Inhibition of Hepatitis B Virus and Hepatitis C Virus in Co-Infected Patients in Chinese Population.

BACKGROUND: Hepatitis B virus (HBV) and hepatitis C virus (HCV) co-infections contributes to a substantial proportion of liver disease worldwide. The aim of this study was to assess the clinical and virological features of HBV-HCV co-infection. METHODS: Demographic data were collected for 3238 high-risk people from an HCV-endemic region in China. Laboratory tests included HCV antibody and HBV serological markers, liver function tests, and routine blood analysis. Anti-HCV positive samples were analyzed for HCV RNA levels and subgenotypes. HBsAg-positive samples were tested for HBV DNA. RESULTS: A total of 1468 patients had chronic HCV and/or HBV infections. Among them, 1200 individuals were classified as HCV mono-infected, 161 were classified as HBV mono-infected, and 107 were classified as co-infected. The HBV-HCV co-infected patients not only had a lower HBV DNA positive rate compared to HBV mono-infected patients (84.1% versus 94.4%, respectively; P < 0.001). The median HCV RNA levels in HBV-HCV co-infected patients were significantly lower than those in the HCV mono-infected patients (1.18[Interquartile range (IQR) 0-5.57] versus 5.87[IQR, 3.54-6.71] Log10 IU/mL, respectively; P < 0.001). Furthermore, co-infected patients were less likely to have detectable HCV RNA levels than HCV mono-infected patients (23.4% versus 56.5%, respectively; P < 0.001). Those HBV-HCV co-infected patients had significantly lower median HBV DNA levels than those mono-infected with HBV (1.97[IQR, 1.3-3.43] versus 3.06[IQR, 2-4.28] Log10 IU/mL, respectively; P < 0.001). The HBV-HCV co-infection group had higher ALT, AST, ALP, GGT, APRI and FIB-4 levels, but lower ALB and total platelet compared to the HBV mono-infection group, and similar to that of the HCV mono-infected group. CONCLUSION: These results suggest that co-infection with HCV and HBV inhibits the replication of both viruses. The serologic results of HBV-HCV co-infection in patients suggests more liver injury compared to HBV mono-infected patients, but is similar to HCV mono-infection.

Factors associated with spontaneous clearance of hepatitis C virus in Chinese population.

Hepatitis C virus (HCV) infections spontaneously clear in approximately 15-45% of infected individuals. Factors which influence spontaneous HCV clearance remain to be identified. The purpose of the present study was to identify variables associated with spontaneous HCV clearance in a referred population of Chinese patients. The prevalence of host, viral, and environmental factors known to influence the outcome of HCV infections was compared in 92 HCV spontaneous clearance subjects and 318 HCV persistent infection subjects. Univariate and multivariate analyses were performed to identify those factors associated with spontaneous HCV clearance. In univariate analysis, female gender, a history of icteric hepatitis, serologic evidence of concurrent HBV infection, and rs12979860 CC genotype were positively associated with spontaneous HCV clearance, while alcohol consumption was negatively associated with clearance. In multivariate analysis, female gender, a history of icteric hepatitis, concurrent HBV infection, and rs12979860 CC genotype remained independent variables associated with spontaneous HCV clearance. Spontaneous HCV clearance is more likely to occur in females, subjects with a history of icteric hepatitis, HBV coinfections, and those with the rs12979860 CC genotype.

[Experimental study on effect of rhizoma drynariae flavone on bone destruction of collagen inducted arthritis rat].

OBJECTIVE: Through establishing the rat model of CIA to evaluate the effect and mechanism of Rhizoma Drynariae Flavone on bone destruction of CIA rat. METHODS: Subcutaneous injection of bovine type II collagen was used to induce Wistar rats to fall ill, and then established the rat model of CIA. The rats whose inflammation scores reached to two points or above were randomly divided into four groups, and were treated accordingly. The effect of Rhizoma Drynariae Flavone on bone destruction was evaluated. RESULTS: At 12 weeks after treatment, bone trabecular area percentage and bone trabecular number in Rhizoma Drynariae Flavone group, Rhizoma Drynariae Flavone-1/2 Etanercept group, Etanercept group was obviously higher than that of sterilization water group (P < 0.05); and the trabecular resolving power of these groups was obviously less than that of sterilization water group (P < 0.05). CONCLUSION: Rhizoma Drynariae Flavone can obviously inhibit inflammation of joint bone destruction of CIA rats,the effect may be related with bone trabecular number reduction and trabecular resolving power increasing.

[Effect of Bushen Qianggu decoction on the proliferation of synovial fibroblasts and expression of PCNA and Bcl-2].

OBJECTIVE: To observe the effects of Bushen Qianggu decoction proliferation and PCNA and Bcl-2 expression. METHODS: Serum containing BQD was made and synovial fibroblasts were separated and cultured and passaged in vitro. Four groups were divided as 20% blank control group, serum containing 20% Tripterygium wilfordii multi-glycosides drug (TWMD), 20% of serum containing high and low of BQD, respectively. Serum containing drugs of different concentration were added into the synovial fibroblasts of the third generation, and then the synovial fibroblasts were cultured continued. The effects of different drugs on synovial fibroblasts and PCNA and Bcl-2 expression were observed. RESULTS: Compared with the control serum, BQD-containing serum promoted the apoptosis of synovial fibroblasts (P < 0.000 1); especially, high dose could inhibit proliferation. The expression of PCNA and Bcl-2 was significantly lower in BQD-containing serum (P < 0.000 1 vs control group). CONCLUSION: BQD can promote the apoptosis of synovial fibroblasts by improving of expression of PCNA and Bcl-2, which may be one of the mechanisms of BQD in preventing and treating osteoporosis of rheumatoid arthritis.

[Effects of Drynaria total flavonoids on apoptosis of osteoblasts mediated by tumor necrosis factor-α].

OBJECTIVE: To investigate the influence of Drynaria total flavonoids on proliferation and apoptosis of osteoblasts in tumor necrosis factor-α (TNF-α)- mediated medium, so as to explore the mechanism of Drynaria total flavonoids in preventing and treating osteoporosis of rheumatoid arthritis. METHODS: Twenty Wistar rats with average weight of (200±20) g were randomly divided into two groups: blank control group and Qianggu capsule (Drynaria total flavonoids) group. Rats in Qianggu capsule group were fed with 75 mg Qianggu capsule daily for continuous 3 d. One hour after the last feed, blood samples were collected. The in vitro experiment of four groups was designed: blank control serum group, Drynaria total flavonoids-containing serum group, blank control serum plus TNF-α group and Drynaria total flavonoids-containing serum plus TNF-α group. Methyl thiazolyl tetrazolium method was used to detect the proliferation of osteoblasts. Flow cytometry was used to detect the apoptosis of osteoblasts and real-time fluorescent quantitative polymerase chain reaction to detect the expressions of Bcl-2 and Bax mRNAs in osteoblasts. RESULTS: Compared with the control serum, Drynaria total flavonoids-containing serum promoted the proliferation and decreased the apoptosis of osteoblasts in TNF-α-mediated inflammatory environment (P<0.05), and increased the ratio of Bcl-2 mRNA to Bax mRNA. CONCLUSION: In TNF-α-mediated inflammatory environment, Drynaria total flavonoids can promote the proliferation and decrease the apoptosis of osteoblasts by improving the ratio of Bcl-2 mRNA to Bax mRNA, which may be one of the mechanisms of Drynaria total flavonoids in preventing and treating osteoporosis of rheumatoid arthritis.