Multifunctional nanocoating for enhanced titanium implant osseointegration.

Preventing bacterial infection and promoting osseointegration are essential for the long-term success of titanium (Ti) implants. In this study, we developed a multifunctional nanocoating on Ti mini-implants to simultaneously address these challenges. The nanocoating consists of self-assembled antimicrobial peptides GL13K and silver nanoparticles, referred to as Ag-GL. Our results showed that the Ag-GL coating did not alter the surface morphology of the mini-implants. Ag-GL coated mini-implants demonstrated a two orders of magnitude reduction in colony-forming unit (CFU) values compared to the noncoated eTi group, resulting in minimal inflammation and no apparent bone destruction in a bacterial infection in vivo model. When evaluating osseointegration properties, micro-CT analysis, histomorphometric analysis, and pull-out tests revealed that the Ag-GL coating significantly enhanced osseointegration and promoted new bone formation in vivo.

Network analysis and experimental approach to investigate the potential therapeutic mechanism of zishen yutai pills on premature ovarian insufficiency.

Background: As society continues to develop, women are more at risk of gonadotoxic substance exposure. Consequently, the incidence of premature ovarian insufficiency (POI) has increased significantly in the past decades. Hormone replacement therapy (HRT) is recommended as the standard treatment to relieve hypoestrogenic symptoms; however, its potential side effects and contraindications have drawn widespread controversy and concern. As such, the Chinese medicine Zishen Yutai Pill (ZSYTP) commonly used for treating miscarriage and menoxenia, is a highly promising alternative drug candidate against POI, however its therapeutic mechanism has not been completely elucidated. Objective: To systematically analyze the potential therapeutic targets of ZSYTP on POI, we combined network pharmacology analysis and molecular docking to predict critical target genes, with experimental validation on POI murine models. Methods: The active compounds of ZSYTP were collected from three online databases, and the candidate targets were predicted based on the chemical structure. The POI-related targets were obtained from four databases. A PPI network was constructed to find the key target genes between ZSYTP and POI, while GO and KEGG enrichment analyses were employed to study the mechanism of ZSYTP against POI. The binding capability of the key co-targets with active components was examined by molecular docking. We used a cyclophosphamide (CTX)-inducible POI mouse model to verify our predictions by histopathological observation, immunohistochemical staining (caspase-3, TUNEL assay), hormone determination (FSH, AMH) and ribonucleic acid sequencing (RNA Seq). Progynova was also used to study the difference between ZSYTP and HRT. Result: We identified 21 target genes as the hub between ZSYTP and POI. The GO and KEGG analyses revealed that the molecular mechanism of ZSYTP against POI were mainly based on the regulation of gene and protein expression. A variety of signaling pathways may be involved in the treatment of ZSYTP against POI, especially PI3K-AKT, HIF-1 and the AGE-RAGE cascades. Docking simulation showed that G1, C1, SR5, and F1 had relatively lower binding energy. In vivo, ZSYTP significantly reversed CTX-induced ovarian damage in follicle number, hormone level and apoptosis, with an overall improved therapeutic effect compared to Progynova. Results from RNA-Seq revealed that the PI3K-AKT, Hippo, AGE-RAGE, and Rap1 signaling pathways and regulation of inflammation, immune response, and lipid metabolism may mediate the protective effects of ZSYTP against POI, which is different than Progynova's mechanism of action. Conclusions: Collectively, this study indicates that ZSYTP could be a highly promising alternative as a non-HRT-based therapy for POI. Its mechanism involves multiple signaling pathways, alleviating ovarian apoptosis and recovering AMH and FSH level. However, the discrepancy between different research techniques highlight the necessity of further experimental verification from other aspects such as translation and posttranslational modification.

CD38 deficiency up-regulated IL-1ß and MCP-1 through TLR4/ERK/NF-κB pathway in sepsis pulmonary injury.

As a disease with high mortality, many cytokines and signaling pathways are associated with sepsis. The pro-inflammatory cytokines and chemokines are participating in the pathogenesis of sepsis, especially in early stage. Moreover, the releases and expressions of cytokines are regulated by numerous signaling pathways, including TLR4/ERK pathway. But despite many studies have expounded the pathogenesis of sepsis and the regulation of cytokines in sepsis, how CD38 influence the expressions of related molecules in sepsis are still unknown. The aim of this study is illuminating the alteration of cytokines and signaling pathways in CD38-/- mice injected with Escherichia coli. Compared with WT mice, E. coli infection results in more severe pulmonary injuries and higher mRNA expressions of cytokines. Compared with E. coli infected WT mice, CD38 knockout leads to aggravated pulmonary injury, increased phosphorylated ERK1/2, p38 and NF-κB p65, and enhanced levels of IL-1ß, iNOS and MCP-1. While compared with E. coli infected CD38-/- mice, TLR4 mutation results in alleviated pulmonary injury, down-regulated phosphorylated ERK1/2 and NF-κB p65, and decreased expressions of IL-1ß and MCP-1. CD38 deficiency increased the expressions of IL-1ß andMCP-1 and aggravated pulmonary injury through TLR4/ERK/NF-κB pathway in sepsis.

CD38 Deficiency Downregulates the Onset and Pathogenesis of Collagen-Induced Arthritis through the NF-κB Pathway.

AIM: The RelB gene plays an important role in guiding the progression of arthritis. We have previously demonstrated that the expression of the RelB gene is decreased significantly in bone marrow DCs of CD38-/- mice. In this study, we demonstrate that the cluster of the differentiation (CD38) gene could be a potentially therapeutic target for autoimmune arthritis. METHOD: Collagen-induced arthritis (CIA) models were generated with both the wild-type (WT) C57BL/6 and CD38-/- mice. The expression of the RelB gene and maturation of bone marrow-derived dendritic cells (DCs) from the WT and CD38-/- mice were detected. Antigen-specific T cell responses, joint damage, and expression of proinflammatory cytokines were assessed. The effects of the Nuclear Factor Kappa B (NF-κB) transcription factor and its mechanisms were characterized. RESULTS: We demonstrated that in CD38-/- mice, the expression of the RelB gene and major histocompatibility complex II (MHC II) was decreased, accompanied with the inhibited T cell reaction in a mixed lymphocyte reaction (MLR) in bone marrow-derived DCs. Compared to the serious degeneration of the cartilage and the enlarged gap of the cavum articular in WT CIA mice, joint pathological changes of the CD38-/- CIA mice revealed marked attenuation, while the joint structures were well preserved. The preserved effects were observed by the inhibition of proinflammatory cytokines and promotion of anti-inflammatory cytokines. Furthermore, decreased phosphorylation of NF-κB was also observed in CD38-/- CIA mice. CONCLUSION: We demonstrate that CD38 could regulate CIA through NF-κB and this regulatory molecule could be a novel target for the treatment of autoimmune inflammatory joint disease.

[In vitro evaluation of the effectiveness of XP-endo Finisher file on smear layer removal after root canal instrumentation].

OBJECTIVE: This study aimed to evaluate the effectiveness of XP-endo Finisher (XPF) file and passive ultrasonic irrigation (PUI) in the smear layer removal of the root canal. METHODS: A total of 60 human single-rooted premolars were selected and decoronated to standardize their canal length to 16 mm. Tooth samples were prepared using a S3 rotary system to prepare root canal with the file size of 3S and then randomly divided into 6 groups according to the final irrigation protocol, as follows: XPF 3 mL of 3% NaOCl for 1 min (group A); XPF 3 mL of 3% NaOCl for 1 min, followed by 4 mL of 17% ethylene diamine tetraacetic acid (EDTA) for 1 min (group B); PUI of 3 mL of 3% NaOCl for 1 min (group C); PUI of 3 mL of 3% NaOCl for 1 min, followed by 4 mL of 17% EDTA for 1 min (group D); 3 mL of 3% NaOCl for 1 min by using a syringe and a 30 G side-vented needle (group E); and 3 mL of 3% NaOCl for 1 min by using a syringe and a 30 G side-vented needle, followed by 4 mL of 17% EDTA for 1 min (group F). After the completion of the root canal preparation, the teeth were split into two longitudinally. The mean numbers of the visible open dentinal tubules in the apical and middle thirds of the root canals were evaluated via scanning electron microscope. RESULTS: The whole surfaces of the root canals in groups A, C, and E were covered by a smear layer. Groups A and C possessed significantly higher number of visible open dentinal tubules than in group E (P<0.05), with statistically insignificant difference between groups A and C (P>0.05). The apical third of the samples in groups B and D and in the middle thirds of canals in group F exhibited a small amount of smear layer, and the dentinal tubules were open or semi-open. The root canal surfaces in the apical third of the samples in group F were covered by a smear layer, and the dentinal tubules were sealed or semisealed. The smear layers in the middle third of the samples in groups B and D were removed, and the dentinal tubules were more visibly open than those of the four other groups (P<0.05). The difference between groups B and D were statistically insignificant (P>0.05). CONCLUSIONS: The difference between XPF and PUI in terms of the smear layer removal of the root canals was insignificant. Hence, XPF, as a new irrigation agitation technique, can aid in improving smear layer removal.

Increased TLR4 Expression Aggravates Sepsis by Promoting IFN-γ Expression in CD38-/- Mice.

Gram-negative bacterial sepsis accounts for up to 50% worldwide sepsis that causes hospital mortality. Acute kidney injury (AKI), a common complication of Gram-negative bacterial sepsis, is caused by Toll-like receptor 4 (TLR4) activation. Lipopolysaccharide (LPS) is an endotoxin in Gram-negative bacteria and is recognized specifically by TLR4, which initiates innate immune response. Also, TLR4 signaling pathway activation is essential in response to LPS infection. CD38 is one of the well-known regulators of innate immunity, whose dysregulation contributes to sepsis. Many studies have proven that an attenuated Gram-positive bacterium induces sepsis in a CD38-blocking model. However, the pathogenesis of Gram-negative bacteria-induced sepsis in a CD38-/- mouse model remains unclear. The aim of this study is to investigate whether kidney injury is still attenuated in a LPS-induced CD38-/- sepsis model and identify the potential mechanism. We assess the severity of kidney injury related to proinflammatory cytokine expressions (IFN-γ, TNF-α, IL-1ß, and IL-6) in WT and CD38-/- mice. Our results showed more aggravated kidney damage in CD38-/- mice than in WT mice, accompanied with an increase of proinflammatory cytokine expression. In addition, compared with CD38-/-TLR4mut mice, we found an increase of TLR4 expression and mRNA expression of these cytokines in the kidney of CD38-/- mice, although only increased IFN-γ level was detected in the serum. Taken together, these results demonstrated that an increased TLR4 expression in CD38-/- mice could contribute to the aggravation of AKI through boosting of the production of IFN-γ.

Effects of menstrual blood­derived stem cells on endometrial injury repair.

The present study aimed to investigate the effects of menstrual blood­derived stem cells (MenSCs) on endometrial injury repair. MenSCs were isolated from human menstrual blood and were cultured in vitro. Flow cytometric analysis of cells in the third generation demonstrated that MenSCs exhibited higher expression levels of cluster of differentiation (CD)90 and lower expression levels of CD146, which suggested that the MenSCs were cultured successfully. A mechanical damage model of unilateral (right) endometrium was established in BALB/c nude mice, which were divided into four groups, Normal, negative control (NC), Model and MenSC. MenSCs transfected with adenovirus­enhanced green fluorescent protein were transplanted into the right uterine cavity of mice in the MenSC and NC groups. The protein expression levels of keratin, vimentin, and vascular endothelial growth factor (VEGF) and the average endometrial thickness were measured by immunohistochemistry; the average optical density of vimentin, VEGF and keratin in the MenSC­treated group was significantly higher compared with the untreated Model group. Fertility tests were performed to determine the pregnancy rate of each group; following endometrial damage in BALB/c nude mice, endometrial thickness was decreased in the Model group, whereas model mice treated with MenSC exhibited increased endometrial thickness and increased the pregnancy rates. Therefore, MenSCs may promote the repair of endometrial lesions in mice by promoting the expression of vimentin, VEGF and keratin.

Flagellin-deficient outer membrane vesicles as adjuvant induce cross-protection of Salmonella Typhimurium outer membrane proteins against infection by heterologous Salmonella serotypes.

Salmonella enteric serovar infections result in high morbidity and mortality worldwide. Cross-protective vaccines are an effective strategy in controlling salmonellosis caused by multiple serotypes. In our previous study, outer membrane vesicles (OMVs) derived from flagellin-deficient Salmonella Typhimurium (S. Typhimurium) were proven effective in mediating cross-protection against infection by multiple Salmonella serotypes; OMVs also exhibit potent adjuvant effects. In this study, we further investigated the adjuvant capacities of flagellin-deficient S. Typhimurium OMVs. Our finding showed that outer membrane proteins (OMPs) in combination with flagellin-deficient S. Typhimurium OMVs could function as adjuvants and invoke stronger humoral, cellular, mucosal, and cross-protective immune responses compared to conventional aluminum (alum). Furthermore, as an adjuvant, OMVs could induce significantly higher cellular immune responses and display enhanced cross-protection for OMPs against wild-type virulent Salmonella Choleraesuis and Salmonella Enteritidis challenge. In summary, OMVs function as a potent adjuvant with the capability of conferring greater cross-protection against infection by multiple Salmonella serotypes, and may be of great value as an effective vaccine adjuvant in enteric diseases.

[Study on an intervention model of "schools without infected students with schistosoma japonica" in heavy endemic areas].

OBJECTIVE: To study an intervention model of "schools without infected students with schistosoma japonica", to control and prevent students from schistosoma infection. METHODS: Twelve primary schools of four heavy endemic counties (districts) with schistosomiasis in the Poyang Lake areas were selected as the study fields, of which, ten schools were the experimental groups, and the other two schools were the control groups by cluster random sampling. All enrolment students were the target population. The baseline survey was carried out in 2005, and an intervention model, "information dissemination + behavior participation + behavior encouragement", was applied in the experiment groups in 2006 - 2008, then the effect of intervention was assessed. RESULTS: Before intervention (2005), the anti-schistosomiasis knowledge awareness rate of experimental and control groups were 14.75% (324/2196) and 16.58% (91/549), and the different was not significant (χ(2) = 1.14, P > 0.05); the rate of accurate attitude of anti-schistosomiasis were 14.71% (323/2196) and 11.84% (65/549) in experimental and control groups, and the difference was not significant (χ(2) = 2.98, P > 0.05); the rate of contacting infected water were 15.44% (18 988/122 976) and 15.03% (4622/30 744) in experimental and control group and the difference was not significant (χ(2) = 3.13, P > 0.05); and the infection rate of schistosomiasis of experiment control groups were 9.65% (212/2196) and 10.56% (58/549), the difference was not significant (χ(2) = 0.41, P > 0.05). After one year intervention (2006), the anti-schistosomiasis knowledge awareness rate of experimental and control groups were 97.79% (2032/2078) and 18.11% (98/541), and the different was significant (χ(2) = 1794.31, P < 0.01); the rate of accurate attitude of anti-schistosomiasis were 99.09% (2059/2078) and 13.49% (73/541) in experimental and control group, and the difference was significant (χ(2) = 2077.45, P < 0.01). After 1 - 3 years intervention (2006 - 2008), there were no any contactors with infected water and infectors with schistosome in students of the experiment group in successive 3 years. While in the control group of the same period, the rate contacting infected water were 16.12% (4884/30 296), 11.11% (3079/27 720) and 12.25% (3451/28 168); the infection rate of schistosomiasis were 8.87% (48/541), 7.47% (37/495) and 7.95% (40/503), respectively. CONCLUSION: The intervention model of health promotion, "information dissemination + behavior participation + behavior encouragement", can effectively control and prevent students from infecting schistosoma japonica in heavy endemic areas with schistosomiasis.

[Establishment and application of school-based health promotion and intervention model of schistosomiasis in lake-type endemic area].

OBJECTIVE: To establish an intervention model of school health promotion, and apply it in developing "schistosomiasis-free schools". METHODS: At the pilot stage, all students of Henghu primary school and Banshan primary school in Xinjian County of Jiangxi Province were selected as experiment group and control group, respectively. A baseline survey covered knowledge and attitude on schistosomiasis control, water contact behaviors and Schistosoma japonicum infection rate. Two health promotion intervention models, i.e. "information communication + training of protection skill + reward & punishment" (model A, 1993-1999) and "information communication + behavior participation + encouragement" (model B, 2000-2007), were implemented in Henghu school. The effect of two models was compared by infection rate. At the application stage, all students of 8 schools in Xinjian County, Nanchang County, and Jinxian County were chosen for evaluation of the effectiveness of Model B with same methods and index. RESULTS: Before intervention there was no significant statistical difference on the passed rate of anti-schistosomiasis knowledge, correct rate of anti-schistosomiasis attitude, frequency of infested water exposure and the infection rate between Henghu and Banshan schools (P > 0.05). In Henghu school, the intervention showed significant effect on the scores of knowledge and attitude after one year (P < 0.01), raised from 9.0% and 55.1% before intervention to 94.4% and 98.9% after intervention, respectively. The frequency of infested water exposure and the infection rate significantly decreased from 14.6% and 13.5% before intervention to 1.9% and 2.3%, respectively (P < 0.01). In 2-7 years after intervention, there were only one or two schistosomiasis cases each year. At the application stage, no schistosomiasis cases were found among Model B target population in two successive years after intervention. CONCLUSION: The practice of Model B can be extended to other schools in endemic area to develop "schistosomiasis-free schools".

The role of health education and health promotion in the control of schistosomiasis: experiences from a 12-year intervention study in the Poyang Lake area.

The objectives of this study were to examine the short-term effects of health education and health promotion in the control of schistosomiasis, and to monitor the long-term impact on re-infection patterns. The study was carried out in six Schistosoma japonicum-endemic villages located in the Poyang Lake area. Three different interventions were implemented, namely (i) health education by means of video tapes, training in prevention of infection, and a 'rewards/punishment' programme for schoolchildren, (ii) promotion of an understanding of schistosomiasis and its control plus training in prevention of infection for women, and (iii) encouragement of compliance with regard to chemotherapy plus training in prevention of infection for men. The 1-year post-intervention follow-up showed that both awareness and appropriate behaviour were strengthened in all three study groups along with a significant increase in the level of knowledge on how to avoid schistosomiasis. For example, the majority of women had abandoned the practice of washing clothes in schistosome-infested water and re-infection rates were sharply reduced as a consequence. In addition, the frequency of water contact among schoolchildren decreased and remained so for the long term. Overall, the approach emphasising health education and health promotion in combination with chemotherapy was highly successful in reducing re-infection rates among inhabitants of S. japonicum-endemic villages and people's compliance with regard to chemotherapy increased significantly over the course of the study.