Molecular detection of multidrug-resistant Pseudomonas aeruginosa of different avian sources with pathogenicity testing and in vitro evaluation of antibacterial efficacy of silver nanoparticles against multidrug-resistant P. aeruginosa.

Pseudomonas aeruginosa (P. aeruginosa) is a serious zoonotic pathogen threaten the poultry industry causing severe economic losses therefor, this study aimed to isolation, phenotypic, molecular identification of P. aeruginosa from different avian sources (chickens, turkey, pigeons, table eggs, and dead in shell chicken embryos), from different Egyptian governorates (Giza, Qalubia, Beheira, El-Minya, and Al-Sharqia) with applying of antibiotic sensitivity test on all P. aeruginosa isolates. Highly resistant isolates (n = 49) were subjected to molecular identification of P. aeruginosa with detection of resistant genes including carbapenemase-encoding genes blaKPC, blaOXA-48, and blaNDM. On the base of molecular results, a highly resistant P. aeruginosa strain was tested for its pathogenicity on day old specific pathogen free (SPF) chicks. Also, in vitro experiment was adopted to evaluate the efficacy of silver nanoparticles (Ag-NPs) against highly antibiotic-resistant P. aeruginosa strains. The overall isolation percentage was from all examined samples were 36.2% (571/1,576) representing 45.2% (532/1,176) from different birds' tissues and 39/400 (9.7%) from total egg samples. Some of isolated strains showed multidrug resistance (MDR) against kanamycin, amoxicillin, amoxicillin-clavulanic acid, neomycin, chloramphenicol, vancomycin, cefotaxime clavulanic acid, lincomycin-spectinomycin, co-trimoxazole, cefoxitin, gentamycin, and doxycycline. These MDR strains were also molecularly positive for ESBL and carbapenemase-encoding genes. MDR strain showed high pathogenicity with histopathological alterations in different organs in challenged birds. Main histopathological lesions were necrosis of hepatocytes, renal tubular epithelium, and heart muscle bundles. The MDR strain showed in vitro sensitivity to Ag-NPs. In conclusion, MDR P. aeruginosa is a serious pathogen causing high morbidity, mortality, and pathological tissue alterations. Ag NPs revealed a promising in vitro antimicrobial sensitivity against MDR P. aeruginosa and further in vivo studies were recommended.

Interleukin-22 Inhibits Apoptosis of Gingival Epithelial Cells Through TGF-ß Signaling Pathway During Periodontitis.

Periodontitis is a chronic inflammatory disease characterized by the destruction of tooth-supporting tissues. The gingival epithelium is the first barrier of periodontal tissue against oral pathogens and harmful substances. The structure and function of epithelial lining are essential for maintaining the integrity of the epithelial barrier. Abnormal apoptosis can lead to the decrease of functional keratinocytes and break homeostasis in gingival epithelium. Interleukin-22 is a cytokine that plays an important role in epithelial homeostasis in intestinal epithelium, inducing proliferation and inhibiting apoptosis, but its role in gingival epithelium is poorly understood. In this study, we investigated the effect of interleukin-22 on apoptosis of gingival epithelial cells during periodontitis. Interleukin-22 topical injection and Il22 gene knockout were performed in experimental periodontitis mice. Human gingival epithelial cells were co-cultured with Porphyromonas gingivalis with interleukin-22 treatment. We found that interleukin-22 inhibited apoptosis of gingival epithelial cells during periodontitis in vivo and in vitro, decreasing Bax expression and increasing Bcl-xL expression. As for the underlying mechanisms, we found that interleukin-22 reduced the expression of TGF-ß receptor type II and inhibited the phosphorylation of Smad2 in gingival epithelial cells during periodontitis. Blockage of TGF-ß receptors attenuated apoptosis induced by Porphyromonas gingivalis and increased Bcl-xL expression stimulated by interleukin-22. These results confirmed the inhibitory effect of interleukin-22 on apoptosis of gingival epithelial cells and revealed the involvement of TGF-ß signaling pathway in gingival epithelial cell apoptosis during periodontitis.

Simulation of Land Use Pattern Based on Land Ecological Security: A Case Study of Guangzhou, China.

The process of rapid urbanization has intensified the conversion of different land use types, resulting in a substantial loss of ecological land and ecological security being threatened. In the context of China's vigorous advocacy of an ecological civilization, it is important to explore future land use patterns under ecological security constraints to promote sustainable development. The insufficient consideration of land ecological security in existing land use pattern simulation studies makes it difficult to effectively promote improvement in the ecological security level. Therefore, we developed a land use simulation framework that integrates land ecological security. Taking the sustainable development of land ecosystems as the core, the land ecological security index (LESI) and ecological zoning (EZ) were determined by the pressure-state-response (PSR) model and the catastrophe progression method (CPM). Natural development (ND) and ecological protection (EP) scenarios were then constructed taking the LESI and EZ into consideration. The CA-Markov model was used to simulate the land use pattern of Guangzhou for 2030 under the two scenarios. The results showed that (1) the study area was divided into four categories: ecological core zone, ecological buffer zone, ecological optimization zone, and urban development zone, with area shares of 37.53%, 31.14%, 16.96%, and 14.37%, respectively. (2) In both scenarios, the construction land around the towns showed outward expansion; compared with the ND scenario, the construction land in the EP scenario decreased by 369.10 km2, and the woodland, grassland, and farmland areas increased by 337.04, 20.80, and 10.51 km2, respectively, which significantly improved the ecological security level. (3) In the EP scenario, the construction land in the ecological core zone, ecological buffer zone, and ecological optimization zone decreased by 85.49, 114.78, and 178.81 km2, respectively, and no new construction land was added in the ecological core zone, making the land use pattern of the EP scenario more reasonable. The results of the study have confirmed that the land use pattern simulation framework integrating land ecological security can effectively predict land use patterns in different future scenarios. This study can provide suggestions and guidance for managers to use in formulating ecological protection policies and preparing territorial spatial planning.

JMJD3 Promotes Porphyromonas gingivalis Lipopolysaccharide-Induced Th17-Cell Differentiation by Modulating the STAT3-RORc Signaling Pathway.

The immune response mediated by Th17 cells is essential in the pathogenesis of periodontitis. Emerging evidence has demonstrated that lipopolysaccharide from Porphyromonas gingivalis (Pg-LPS) could promote Th17-cell differentiation directly, while the downstream signaling remains elusive. This study was aimed to explore the role of JMJD3 (a JmjC family histone demethylase) and signal transducers and activators of transcription 3 (STAT3) in Th17-cell differentiation triggered by Pg-LPS and clarify the interaction between them. We found that the expression of JMJD3 and STAT3 was significantly increased under Th17-polarizing conditions. Pg-LPS could promote Th17-cell differentiation from CD4+ T cells, with an increased expression of JMJD3 and STAT3 compared to the culture without Pg-LPS. The coimmunoprecipitation results showed that the interactions of JMJD3 and STAT3, STAT3 and retinoid-related orphan nuclear receptor γt (RORγt) were enhanced following Pg-LPS stimulation during Th17-cell differentiation. Further blocking assays were performed and the results showed that inhibition of STAT3 or JMJD3 both suppressed the Th17-cell differentiation, JMJD3 inhibitor could reduce the expression of STAT3 and p-STAT3, while JMJD3 expression was not affected when STAT3 was inhibited. Taken together, this study found that JMJD3 could promote Pg-LPS induced Th17-cell differentiation by modulating the STAT3-RORc signaling pathway.

The Compound Effect of Spatial and Temporal Resolutions on the Accuracy of Urban Flood Simulation.

Flood disaster is one of the critical threats to cities. With the intellectualization tendency of Industry 4.0, refined urban flood models can effectively reproduce flood inundation scenarios and support the decision-making on the response to the flood. However, the spatiotemporal variability of rainfall and the spatial heterogeneity of the surface greatly increase the uncertainties in urban flood simulations. Therefore, it is crucial to account for spatiotemporal variability of rainfall events and grids of the model as accurately as possible to avoid misleading simulation results. This study aims to investigate the effect of temporal resolutions of rainfall and spatial resolutions of the model on urban flood modeling in small urban catchments and to explore a proper combination of spatiotemporal schemes. The IFMS Urban (integrated flood modeling system, urban) is used to construct a one-dimension and two-dimension coupled urban flood model in the typical inundated area in Dongguan, China. Based on five temporal resolutions of rainfall input and four spatial resolutions, the compound effect of spatiotemporal resolutions on the accuracy of urban flood simulations is systematically analyzed, and the variation characteristics are investigated. The results show that the finer the temporal resolution is, the higher the simulation accuracy of the maximum inundated water depth. Considering the spatial resolution, as the spatial grid becomes smaller, the relative error of the maximum inundated water depth decreases, but it also shows some nonlinear characteristics. Therefore, the smaller grid does not always mean a better simulation. The spatial resolution has a greater impact on the flood simulation accuracy than the temporal resolution. The simulation performance reaches the best when the grid interval is 100 m and the rainfall input interval is 5 min, 10 min, or 15 min. Affected by other factors such as terrain slope, the simulation accuracies under different spatiotemporal resolutions present complex nonlinear characteristics. The mechanisms of the compound effect of the spatiotemporal resolutions on the model simulation and the effect of underlying surface and topography on model simulation will be the focus of in-depth exploration for the future urban flood model.

The association of periodontal diseases and Sjogren's syndrome: A systematic review and meta-analysis.

Background: The relationship between periodontal diseases and Sjogren's syndrome were found inconsistent in current studies. Our objective is to clarify the relationship between periodontal diseases and Sjogren's syndrome. Methods: A systematic review was performed and reported according to Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA). Electronic databases (EMBASE, PubMed, Web of Science, and Cochrane Library, from inceptions until 24 November 2021) were searched. The Newcastle-Ottawa Scale (NOS) and Agency for Healthcare Research and Quality (AHRQ) were applied to evaluate the quality of studies. Quality assessment of the certainty of evidence was performed based on the Grading of Recommendations Assessment, Development, and Evaluation (GRADE) guidelines. When the output is the ratio, Odds ratio (OR) of periodontal diseases with Sjogren's syndrome were calculated. When the output is the mean, weighted mean difference (WMD) of periodontal diseases with Sjogren's syndrome was calculated. We conducted meta-analysis and estimated the pool sensitivity. Begg's test was used to test the possibility of publication bias. We also carried out meta-regression to clarify the source of heterogeneity (I2 > 50%). Finally, we performed a trial sequential analysis (TSA) to identify the false positive or false negative outcomes that might occur during repeated updates. Results: 21 studies were included in this systematic review, with a total of 11435 subjects. Meta-analysis of 5 studies showed that there is a positive correlation between periodontitis and Sjogren's syndrome (OR = 2.12, 95% CI = 1.43-3.17; 5 studies, 6927 participants; low certainty of evidence). Meta-analysis of 16 studies showed that the periodontal condition of patients with Sjogren's syndrome was worse compared with the control group, and the scores of clinical periodontal parameters were relatively high. Conclusion: Sjogren's syndrome patients seem to be more likely to be diagnosed with periodontal diseases. However, our results should be interpreted with caution considering the high heterogeneity. Systematic review registration: [https://www.crd.york.ac.uk/prospero/], identifier [CRD42021261322].

Porphyromonas gingivalis lipopolysaccharide promotes T-hel per17 cell differentiation by upregulating Delta-like ligand 4 expression on CD14+ monocytes.

BACKGROUD: To investigate the effect and mechanism of Porphyromonas gingivalis (P. gingivalis) lipopolysaccharide (LPS) on Th17 cell differentiation mediated by CD14+ monocytes. METHODS: P. gingivalis LPS-activated CD14+ monocytes were co-cultured with CD4+T cells in different cell ratios. An indirect co-culture system was also established using transwell chambers. Furthermore, anti- Delta-like ligand 4 (Dll-4) antibody was used to investigate the role of Dll-4 in Th17 cell response. The mRNA expression was analyzed using quantitative reverse transcription-polymerase chain reaction, and secreted cytokines in culture supernatant were detected using enzyme-linked immunosorbent assay. Flow cytometry was used to determine the frequencies of Th17 cells. IL-17 protein expression levels were determined using western blotting assay. RESULTS: P. gingivalis LPS increased the expressions of interleukin (IL)-1ß, IL-6, IL-23 and transforming growth factor (TGF)-ß in CD14+ monocytes. Th17 cell frequency upregulated, which is not solely cytokine-dependent but rather requires cell-cell contact with activated monocytes, particularly in the 1:10 cell ratio. Furthermore, P. gingivalis LPS increased t he expression of Dll-4 on CD14+ monocytes, whereas the anti- Dll-4 a ntibody decreased the response of Th17 cells. The results suggest that P. gingivalis LPS enhances Th17 cell response via Dll-4 upregulation on CD14+ monocytes.

Effect of interleukin-22 on osteogenic differentiation and the osteoclastogenic response of human periodontal ligament fibroblasts in vitro.

BACKGROUND: Interleukin-22 (IL-22) exerts extensive biological effects, playing both protective and pathological roles in autoimmune and infectious diseases. However, the specific role and mechanism of IL-22 in the pathogenesis of periodontitis have not been clarified. The aim of this study was to analyze the possible roles of IL-22 in the osteoclastogenesis and osteogenesis of periodontitis. METHODS: Human periodontal ligament fibroblasts (hPDLFs) were treated with IL-22 and/or lipopolysaccharide from Porphyromonas gingivalis (Pg-LPS), and the mRNA and protein expression of RANKL and OPG were measured by qRT-PCR and Western blotting, respectively. Western blotting was also used to examine the phosphorylated and total protein expression of MAPK signaling molecules. The role of the MAPK pathway in osteoclastogenesis marker expression was further confirmed by inhibition assays. For osteogenic assays, the mRNA expression of osteoblastic markers was quantified by qRT-PCR, the alkaline phosphatase (ALP) activity of hPDLFs was measured by an ALP assay, and the mineralized nodules formed by hPDLFs were determined by Alizarin Red S staining. RESULTS: IL-22 promoted the expression of RANKL in hPDLFs via the MAPK signaling pathway and further upregulated RANKL expression together with Pg-LPS via the p38 MAPK pathway. IL-22 could enhance the ALP activity and mineralized nodule formation of hPDLFs in the early period of osteogenic induction, while exhibiting no profound effect on the expression of osteoblastic markers. CONCLUSION: IL-22 plays regulatory roles in bone homeostasis, and it is likely to contribute to osteoclastogenesis as a proinflammatory cytokine in the pathogenesis of periodontitis.

Research on the roles of genes coding ATP-binding cassette transporters in Porphyromonas gingivalis pathogenicity.

Porphyromonas gingivalis, as a major pathogen of periodontitis, could rapidly adhere to and invade host gingival epithelial cells (GECs) for the induction of infection. One ATP-binding cassette (ABC) transporter gene was found to be upregulated during this infection process, however, the molecular mechanisms remain unclear. In this study, we systemically investigated the messenger RNA level changes of all ABC transporter family genes in P. gingivalis while being internalized within GECs by real-time polymerase chain reaction. We identified that two ABC transporter genes, PG_RS04465 (PG1010) and PG_RS07320 (PG1665), were significantly increased in P. gingivalis after coculturing with GECs. Mutant strains with knockout (KO) of these two genes were generated by homogenous recombination. PG_RS04465 and PG_RS07320 KO mutants showed no change in the growth of bacteria per se. Knockdown of PG_RS07320, but not PG_RS04465, caused decreased endotoxin level in the bacteria. In contrast, both mutant strains showed decreased Arg- and Lys-gingipains activities, with significantly reduced adhesion and invasion capabilities. Secreted interleukin-1ß (IL-1ß) and IL-6 levels in GECs cocultured with PG_RS04465 or PG_RS07320 KO mutants were also decreased, whereas, only the cells cocultured with PG_RS07320 KO mutants showed significant decrease. In addition, virulence study using mouse revealed that both KO mutant strains infection caused less mouse death than wild-type strains, showing reduced virulence of two KO strains. These results indicated that ABC transporter genes PG_RS04465 and PG_RS07320 are positive regulators of the virulence of P. gingivalis.

HIV incidence and cohort retention among men who have sex with men in Hangzhou, China: A prospective cohort study.

Prospective cohort studies have been conducted to estimate HIV incidence among men who have sex with men (MSM) in first-line megacities cities (>10 million residents) in China, but few in the second-line large- or middle-size cities. This study was to investigate HIV incidence and cohort retention among MSM in a second-line city Hangzhou in eastern China.A total of 523 HIV-seronegative MSM were recruited during September 2014 to September 2015, and were followed up prospectively at 3, 6, 9, and 12 months. Questionnaire interviews were conducted, and laboratory tests were performed to evaluate baseline syphilis infection and HIV seroconversions. Chi-square test and logistic regression model were used to identify factors associated with cohort retention rate and syphilis prevalence.Of 523 participants, 137 (26.2%) completed 6-month follow-up, and use of Internet for recruiting study participants (vs other recruitments: adjusted odds ratio [AOR] = 0.5; 95% confidence interval [CI]: 0.3-0.8) and being homosexual (vs heterosexual or bisexual: AOR = 0.6; 95% CI: 0.4-0.9) were associated with lower cohort retention. The overall HIV incidence during 12 months of follow-up was 6.6 per 100 person-years (95% CI: 3.4-9.8/100 PY). The prevalence of syphilis at baseline was 6.5% (95% CI: 4.4%-8.6%), and disclosing sexual orientation (AOR = 0.4, 95% CI: 0.2-0.9) was associated with lower risk of syphilis infection.HIV is spreading rapidly among MSM in the second-line Chinese city. Effective interventions are needed to target this population in both first-line megacities and second-line large and middle-size cities.

Porphyromonas gingivalis lipopolysaccharide promotes T- helper 17 cell differentiation from human CD4+ naïve T cells via toll-like receptor-2 in vitro.

OBJECTIVES: The persistence of T-helper 17 (Th17) cells has been shown to support chronic inflammation and mediate tissue destruction in periodontitis. However, little is known regarding the underlying mechanisms that regulate Th17 cell differentiation in the periodontal inflammatory context. The objective of this study was to explore the possible effect and mechanism of lipopolysaccharide (LPS) from Porphyromonas gingivalis on Th17 cell differentiation. METHODS: Activated human CD4+CD45RA+ naïve T cells were stimulated with different doses of LPS from virulent and avirulent P. gingivalis strains combined with Th17 driven cytokines in vitro. Flow cytometry was used to analyze the differentiation ratio of Th17 cells. IL-17A protein expression and IL-17, retinoid-related orphan receptor C (RORC) and toll-like receptor 2 (TLR2) mRNA transcription were analysed by ELISA and real-time qPCR, respectively. The role of TLR2 in Th17 cell differentiation was further confirmed by TLR2 blocking assay. RESULTS: LPS from P. gingivalis (Pg-LPS) up-regulated Th17 cell differentiation ratios, expression of IL-17 and RORC mRNA, and IL-17 concentration in culture supernatant, with 0.1 µg/mL LPS from the virulent P. gingivalis strain being the most effectively. Furthermore, Pg-LPS also up-regulated expression of TLR2 on T cells during Th17 differentiation, and the differentiation was attenuated by treatment with TLR2 antibody. CONCLUSIONS: These results suggest that Pg-LPS promotes Th17 cell differentiation in vitro, and TLR2 signalling may be involved in this process. LPS from the virulent P. gingivalis strain up-regulated Th17 cell differentiation more effectively, which may be associated with the pathogenicity of different P. gingivalis strains.

HIV care continuum among newly diagnosed student and non-student youths between 2012 and 2016 in Hangzhou, China.

BACKGROUND: Youths are disproportionally affected by the human immunodeficiency virus (HIV) infection. We aimed to assess anti-retroviral therapy (ART) initiation and viral suppression rates among student and non-student youths in Hangzhou, China. METHODS: Data were taken from the Chinese HIV/acquired immune deficiency syndrome Comprehensive Response Information Management System. Youths aged 15 to 24 years who were newly diagnosed with HIV between 2012 and 2016 and were living in Hangzhou were included in the study. Comparisons between student and non-student youths were made for ART initiation within 30 days, 90 days, and 12 months of HIV diagnosis, and the viral suppression rate at 12 months of HIV diagnosis and at 12 months of ART initiation. RESULTS: A total of 707 cases met inclusion criteria, 29.6% of which were students and 70.4% were non-student youths. The student group had a higher proportion of ART initiation compared with the non-student group within 30 days of diagnosis (45.5% vs. 37.0%, P = 0.044), and a slightly higher but not statistically significant proportion at 90 days (67.0% vs. 62.7%), and 12 months (83.7% vs. 78.5%) of HIV diagnosis. ART initiation within 30 days improved from <15% in 2012 to over 65% in 2016 in both groups, and ART initiation within 90 days improved from <30% in 2012 to >90% in 2016. A smaller proportion of students experienced viral suppression compared with the non-student group (9.6% vs. 17.1%, P = 0.011) at 12 months after HIV diagnosis, but the suppression rate was similar at 12 months of ART initiation (69.9% vs. 71.1%, P = 0.743). CONCLUSIONS: ART initiation in both student and non-student youths has significantly improved between 2012 and 2016. However, the viral suppression rate remained unacceptably low at 12 months of HIV diagnosis in both student and non-student groups. Specific intervention strategies must be taken to address this challenge.

New types of drug use and risks of drug use among men who have sex with men: a cross-sectional study in Hangzhou, China.

BACKGROUND: The use of new types of drugs has become more common among men who have sex with men (MSM). The aim of this study was to describe the patterns of the use of new types of drugs, such as methamphetamine, ketamine, ecstasy, and rush poppers, and to examine the factors associated with drug use and HIV infection among MSM in Hangzhou, China. METHODS: This cross-sectional study was conducted between August 2015 and April 2016. We used snowball sampling to recruit MSM; participants were recruited from voluntary counseling and testing centers, baths, bars, Blued (an app for the gay community), QQ groups, clubs, and other types of venues. MSM were included if their previous HIV test results were negative or unknown, or they had not been tested for HIV. MSM were excluded if they were known to be HIV positive before the survey. Face-to-face questionnaires were conducted and a venous blood specimen was drawn from each participant following the interview. RESULTS: In total, 555 MSM were included; 18.2% (101/555) of the participants had used new types of drugs in the past 3 months. Among the users, 65.3% used single-use rush poppers, while the remainder used ketamine, methamphetamine, ecstasy, or other mixed combinations of drugs. The HIV positivity rate was 14.8% (82/555). Factors associated with increased odds of using new types of drugs in the past 3 months were higher education levels (adjusted odds ratio [AOR] 4.45, 95% confidence interval [CI] 2.12-9.37), having multiple sexual partners (AOR 1.76, 95 CI 1.02-3.05), alcohol use before sexual intercourse (AOR 33.44, 95% CI 10.80-103.50), and seeing friends using new types of drugs. CONCLUSION: We revealed the widespread use of new types of drugs, as well as a high diagnosis rate of new HIV infection, among MSM in Hangzhou. The use of new types of drugs was associated with an increased number of sexual partners among MSM; the high-risk sexual behaviors increased the risk of HIV infection. Attention should be given to the use of new types of drugs in MSM, and supervision programs should be strengthened to combat drug use.

Quantitative Evaluation of MMP-9 and TIMP-1 Promoter Methylation in Chronic Periodontitis.

In this study, we investigated the promoter DNA methylation (DNAm) status of the MMP-9 and TIMP-1 genes in patients with chronic periodontitis to evaluate disease progression. Using pyrosequencing technology, DNAm levels of MMP-9 and TIMP-1 CpG islands were measured in 88 chronic periodontitis patients and 15 healthy controls. We found a positive correlation between methylation levels of MMP-9 CpG islands and the severity of chronic periodontitis. Methylated CpG islands were also closely associated with the duration of chronic periodontitis. Moreover, female patients exhibited lower methylation levels of MMP-9 but higher methylation levels of TIMP-1 compared with male patients, and the methylation levels of TIMP-1 gradually decreased with age. The findings of gender disparity in the DNAm of MMP-9 and TIMP-1 genes provide novel insights into chronic periodontitis.

Quercetin Inhibits LPS-Induced Inflammation and ox-LDL-Induced Lipid Deposition.

Aberrant activation of inflammation and excess accumulation of lipids play crucial role in the occurrence and progression of atherosclerosis (AS). Quercetin (QCT) has been tested effectively to cure AS. It is widely distributed in plant foods and has been proved to have potential antioxidative and anticancer activities. However, the underlying molecular mechanisms of OCT in AS are not completely understood. In the present study, we stimulated murine RAW264.7 cells with lipopolysaccharide (LPS) or oxidized low-density lipoproteins (ox-LDL) to mimic the development of AS. The data show that QCT treatment leads to an obvious decrease of multiple inflammatory cytokines in transcript level, including interleukin (IL)-1α, IL-1ß, IL-2, IL-10, macrophage chemoattractant protein-1 (MCP-1), and cyclooxygenase-2 (COX-2) induced by LPS. Moreover, expressions of other factors that contribute to the AS development, such as matrix metalloproteinase-1 (MMP-1) and suppressor of cytokine signaling 3 (SOCS3) induced by LPS are also downregulated by QCT. Furthermore, we found that QCT suppressed LPS-induced the phosphorylation of STAT3. Meanwhile, QCT could ameliorate lipid deposition and overproduction of reactive oxygen species induced by ox-LDL, and block the expression of lectin-like oxidized LDL receptor-1 (LOX-1) in cultured macrophages. Taken together, our data reveal that QCT has obvious anti-inflammatory and antioxidant virtues and could be a therapeutic agent for the prevention and treatment of AS.

Dual role of the integrated stress response in medulloblastoma tumorigenesis.

In response to endoplasmic reticulum (ER) stress, activation of pancreatic ER kinase (PERK) coordinates an adaptive program known as the integrated stress response (ISR) by phosphorylating translation initiation factor 2α (eIF2α). Phosphorylated eIF2α is quickly dephosphorylated by the protein phosphatase 1 and growth arrest and DNA damage 34 (GADD34) complex. Data indicate that the ISR can either promote or suppress tumor development. Our previous studies showed that the ISR is activated in medulloblastoma in both human patients and animal models, and that the decreased ISR via PERK heterozygous deficiency attenuates medulloblastoma formation in Patched1 heterozygous deficient (Ptch1+/-) mice by enhancing apoptosis of pre-malignant granule cell precursors (GCPs) during cell transformation. We showed here that GADD34 heterozygous mutation moderately enhanced the ISR and noticeably increased the incidence of medulloblastoma in adult Ptch1+/- mice. Surprisingly, GADD34 homozygous mutation strongly enhanced the ISR, but significantly decreased the incidence of medulloblastoma in adult Ptch1+/- mice. Intriguingly, GADD34 homozygous mutation significantly enhanced pre-malignant GCP apoptosis in cerebellar hyperplastic lesions and reduced the lesion numbers in young Ptch1+/- mice. Nevertheless, neither GADD34 heterozygous mutation nor GADD34 homozygous mutation had a significant effect on medulloblastoma cells in adult Ptch1+/- mice. Collectively, these data imply the dual role of the ISR, promoting and inhibiting, in medulloblastoma tumorigenesis by regulating apoptosis of pre-malignant GCPs during the course of malignant transformation.

PERK Activation Promotes Medulloblastoma Tumorigenesis by Attenuating Premalignant Granule Cell Precursor Apoptosis.

Evidence suggests that activation of pancreatic endoplasmic reticulum kinase (PERK) signaling in response to endoplasmic reticulum stress negatively or positively influences cell transformation by regulating apoptosis. Patched1 heterozygous deficient (Ptch1(+/-)) mice reproduce human Gorlin's syndrome and are regarded as the best animal model to study tumorigenesis of the sonic hedgehog subgroup of medulloblastomas. It is believed that medulloblastomas in Ptch1(+/-) mice results from the transformation of granule cell precursors (GCPs) in the developing cerebellum. Here, we determined the role of PERK signaling on medulloblastoma tumorigenesis by assessing its effects on premalignant GCPs and tumor cells. We found that PERK signaling was activated in both premalignant GCPs in young Ptch1(+/-) mice and medulloblastoma cells in adult mice. We demonstrated that PERK haploinsufficiency reduced the incidence of medulloblastomas in Ptch1(+/-) mice. Interestingly, PERK haploinsufficiency enhanced apoptosis of premalignant GCPs in young Ptch1(+/-) mice but had no significant effect on medulloblastoma cells in adult mice. Moreover, we showed that the PERK pathway was activated in medulloblastomas in humans. These results suggest that PERK signaling promotes medulloblastoma tumorigenesis by attenuating apoptosis of premalignant GCPs during the course of malignant transformation.

Risk factors for HIV and syphilis infection among male sex workers who have sex with men: a cross-sectional study in Hangzhou, China, 2011.

OBJECTIVE: To investigate the prevalence and risk factors of HIV and syphilis infection among men who have sex with men (MSM) in male sex workers (MSW). DESIGN: Cross-sectional survey. SETTING: Hangzhou, China. PARTICIPANTS: 259 MSW in MSM were recruited by respondent-driven sampling from May 2011 to December 2011.The inclusion criteria were: (1) age ≥18 years; (2) engaging in sex with men in the previous year and (3) willing to cooperate in the implementation of the study. OUTCOME MEASURES: HIV-related knowledge, high-risk behaviour and condom use. RESULTS: Among these MSW in MSM, 23.2% were infected with HIV and/or syphilis, 8.9% were infected only with HIV, 12.7% only with syphilis and 1.5% with HIV/syphilis co-infection; 96.6% sold sex to males, 8.9% bought sex from males and 15.4% sold sex to females; 49.0% had non-commercial sex behaviours with males and 24.3% with females. The rate of condom use while having commercial sex with clients was 86.9% and 53.3% (selling anal and oral sex to males, respectively), 95.5% (buying sex from males) and 77.5% (selling sex to females), respectively. Regarding their non-commercial sex behaviour, the rate of condom use was 77.2% (with males) and 49.2% (with females), respectively. Multivariate analysis showed that age >30 years (OR 1.055; 95% CIs 1.015 to 1.095) and having ≥10 non-commercial male sex partners (OR, 1.573; 95% CI 1.018 to 2.452) were significantly associated with HIV/syphilis infection, while heterosexuality (OR, 0.238; 95% CI 0.066 to 0.855) was significantly associated with a low HIV/syphilis infection rate. CONCLUSIONS: The MSW in MSM population in Hangzhou has a high prevalence of HIV/syphilis infection, poor perceived risks of HIV and more engagement in unsafe sex with its clients and partners, in addition to a low rate of condom use. These risk factors may account for their relatively high infection rate of HIV/syphilis.

Attenuation of Porphyromonas gingivalis oral infection by α-amylase and pentamidine.

The Porphyromonas gingivalis bacterium is one of the most influential pathogens in oral infections. In the current study, the antimicrobial activity of α-amylase and pentamidine against Porphyromonas gingivalis was evaluated. Their in vitro inhibitory activity was investigated with the agar overlay technique, and the minimal inhibitory and bactericidal concentrations were determined. Using the bactericidal concentration, the antimicrobial actions of the inhibitors were investigated. In the present study, multiple techniques were utilized, including scanning electron microscopy (SEM), general structural analysis and differential gene expression analysis. The results obtained from SEM and bactericidal analysis indicated a notable observation; the pentamidine and α-amylase treatment destroyed the structure of the bacterial cell membranes, which led to cell death. These results were used to further explore these inhibitors and the mechanisms by which they act. Downregulated expression levels were observed for a number of genes coding for hemagglutinins and gingipains, and various genes involved in hemin uptake, chromosome replication and energy production. However, the expression levels of genes associated with iron storage and oxidative stress were upregulated by α-amylase and pentamidine. A greater effect was noted in response to pentamidine treatment. The results of the present study demonstrate promising therapeutic potential for α-amylases and pentamidine. These molecules have the potential to be used to develop novel drugs and broaden the availability of pharmacological tools for the attenuation of oral infections caused by Porphyromonas gingivalis.

Effects of cordycepin on HepG2 and EA.hy926 cells: Potential antiproliferative, antimetastatic and anti-angiogenic effects on hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is a hypervascular tumor and accumulating evidence suggests that angiogenesis plays an important role in HCC development. Cordycepin, also known as 3'-deoxyadenosine, is a derivative of adenosine, and numerous cellular enzymes cannot differentiate the two. The aim of the present study was to determine whether cordycepin regulates proliferation, migration and angiogenesis in a human umbilical vein endothelial cell line (EA.hy926) and in a hepatocellular carcinoma cell line (HepG2). MTT was used to assess cell proliferation. Apoptosis was analyzed by flow cytometry (propidium iodide staining). Transwell and wound healing assays were used to analyze the migration and invasion of HepG2 and EA.hy926 cells. Angiogenesis in EA.hy926 cells was assessed using a tube formation assay. Cordycepin strongly suppressed HepG2 and EA.hy926 cell proliferation in a dose- and time-dependent manner. Cordycepin induced EA.hy926 cell apoptosis in a dose-dependent manner (2,000 µg/ml: 50.20±1.55% vs. 0 µg/ml: 2.62±0.19%; P<0.01). Cordycepin inhibited EA.hy926 cell migration (percentage of wound healing area, 2,000 µg/ml: 3.45±0.29% vs. 0 µg/ml: 85.48±0.84%; P<0.05), as well as tube formation (total length of tubular structure, 1,000 µg/ml: 107±39 µm vs. 0 µg/ml: 936±56 µm; P<0.05). Cordycepin also efficiently inhibited HepG2 cell invasion and migration. High-performance liquid chromatography analysis of the cytosol from EA.hy926 cells showed that cordycepin was stable for 3 h. In conclusion, cordycepin not only inhibited human HepG2 cell proliferation and invasion, but also induced apoptosis and inhibited migration and angiogenesis in vascular endothelial cells, suggesting that cordycepin may be used as a novel anti-angiogenic therapy in HCC.