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Background To validate tumor volume-based imaging markers for predicting local recurrence-free survival (LRFS) in locoregionally advanced nasopharyngeal carcinoma patients, who underwent induction chemotherapy followed by definitive intensity-modulated radiotherapy. Methods We enrolled 145 patients with stage III–IVA nasopharyngeal carcinoma in this retrospective study. Pre-treatment tumor volume (Vpre) and late-course volume (LCV) were measured based on the MRIs scanned before treatment and during the first 3 days in the sixth week of radiotherapy, respectively. The volume regression rate (VRR) was calculated according to Vpre and LCV. Receiver operating characteristic (ROC) curves were used to identify the cut-off best separating patient subgroups in assessing the prognostic value of Vpre, LCV and VRR. The Kaplan–Meier method was used for survival analysis. Prognostic analyses were performed using univariate and multivariate COX proportional hazard models. Results The LCV was 5.3 ± 0.5 (range 0–42.1) cm3; The VRR was 60.4 ± 2.2% (range 2.9–100.0). The median follow-up period was 36 months (range 6–98 months). The cut-off value of LCV determined by the ROC was 6.8 cm3 for LRFS prediction (sensitivity 68.8%; specificity 79.8%). The combination of LCV and VRR for LRFS prediction (AUC = 0.79, P P = 0.002, 95% CI 0.60–0.88) and Vpre (AUC = 0.71, P = 0.007, 95% CI 0.56–0.85) are better than T category (AUC = 0.64, P = 0.062, 95% CI 0.50–0.79) alone. Patients with LCV ≤ 6.8 cm3 had significantly longer LRFS (P P P = 0.005) than those with LCV > 6.8 cm3. Multivariate Cox regression showed LCV was the only independent prognostic factor for local control (HR = 7.80, 95% CI 2.69–22.6, P Conclusions LCV is a promising prognostic factor for local control and chemoradiosensitivity in patients with locoregionally advanced NPC. The LCV, and the combination of LCV with VRR are more robust predictors for patient survival than T category.

Background and purpose Adiponectin (APN) is an adipokine secreted from adipocytes that binds to APN receptors AdipoR1 and AdipoR2 and exerts an anti-inflammatory response through mechanisms not fully understood. There is a need to develop small molecules that activate AdipoR1 and AdipoR2 and to be used to inhibit the inflammatory response in lipopolysaccharide (LPS)-induced endotoxemia and other inflammatory disorders. Experimental approach We designed 10 new structural analogues of an AdipoR agonist, AdipoRon (APR), and assessed their anti-inflammatory properties. Bone marrow-derived macrophages (BMMs) and peritoneal macrophages (PEMs) were isolated from mice. Levels of pro-inflammatory cytokines were measured by reverse transcription and real-time quantitative polymerase chain reaction (qRT-PCR), enzyme-linked immunosorbent assay (ELISA) and microarray in LPS-induced endotoxemia mice and diet-induced obesity (DIO) mice in which systemic inflammation prevails. Western blotting, immunohistochemistry (IHC), siRNA interference and immunoprecipitation were used to detect signalling pathways. Key results A novel APN receptor agonist named adipo anti-inflammation agonist (AdipoAI) strongly suppresses inflammation in DIO and endotoxemia mice, as well as in cultured macrophages. We also found that AdipoAI attenuated the association of AdipoR1 and APPL1 via myeloid differentiation marker 88 (MyD88) signalling, thus inhibiting activation of nuclear factor kappa B (NF-κB), mitogen-activated protein kinase (MAPK) and c-Maf pathways and limiting the production of pro-inflammatory cytokines in LPS-induced macrophages. Conclusion and implications AdipoAI is a promising alternative therapeutic approach to APN and APR to suppress inflammation in LPS-induced endotoxemia and other inflammatory disorders via distinct signalling pathways.

Objectives The study aimed to explore the effects of mouth breathing and hypoxia on the condyle of temporomandibular joint (TMJ) via two animal models. Methods 24 four-week-old rats were randomly separated into three groups, consisting of eight control rats, eight intermittent hypoxia (IH) rats, and eight intermittent nasal obstruction (INO) rats. We use the IH model and the INO model to simulate children suffering from hypoxia and mouth breathing. After 16 days, the condyle of TMJ and surrounding white adipose tissue (WAT) and skeletal muscle tissue were obtained for further staining and qRT-PCR. Finally, RNA-seq was used to verify the results. Results The intermittent hypoxia cannot significantly change the overall structure in the cause of short-term hypoxia stimulation, but the intermittent nasal obstruction can alter the condyle, WAT, and muscle, while also introducing noticeable structural changes in tissue hypoxia and macrophage infiltration. Sequencing data verified these findings and also suggested that this process might involve the Hif-1α/Vegf axis. Conclusions Our findings reveal the very early structural impact of mouth breathing on condyle reconstruction in rat models, and hypoxia does not induce evident alteration on condyle. However, since these results are mainly focused on rats, further studies are needed to understand its effects on humans.

Macrophages exhibit diverse functions within various tissues during the inflammatory response, and the physical properties of tissues also modulate the characteristics of macrophages. However, the underlying N6-methyladenosine (m

As a precursor to type 2 diabetes mellitus (T2D), obesity adversely alters bone cell functions, causing decreased bone quality. Currently, the mechanisms leading to alterations in bone quality in obesity and subsequently T2D are largely unclear. Emerging evidence suggests that long noncoding RNAs (lncRNAs) participate in a vast repertoire of biological processes and play essential roles in gene expression and posttranscriptional processes. Mechanistically, the expression of lncRNAs is implicated in pathogenesis surrounding the aggregation or alleviation of human diseases. To investigate the functional link between specific lncRNA and obesity-associated poor bone quality and elucidate the molecular mechanisms underlying the interaction between the two, we first assessed the structure of the bones in a diet-induced obese (DIO) mouse model. We found that bone microarchitecture markedly deteriorated in the DIO mice, mainly because of aberrant remodeling in the bone structure. The results of in vitro mechanistic experiments supported these observations. We then screened mRNAs and lncRNAs from DIO bones and functionally identified a specific lncRNA, Gm15222. Further analyses demonstrated that Gm15222 promotes osteogenesis and inhibits the expression of adipogenesis-related genes in DIO via recruitment of lysine demethylases KDM6B and KDM4B, respectively. Through this epigenetic pathway, Gm15222 modulates histone methylation of osteogenic genes. In addition, Gm15222 showed a positive correlation with the expression of a neighboring gene, BMP4. Together, the results of this study identified and provided initial characterization of Gm15222 as a critical epigenetic modifier that regulates osteogenesis and has potential roles in targeting the pathophysiology of bone disease in obesity and potential T2D.

BACKGROUND AND OBJECTIVE: Type 2 diabetes (T2D)-associated periodontitis is severe and refractory in many cases. Considered an inflammatory disease, T2D predisposes to periodontitis by increasing whole-body inflammation. One mechanism of increased inflammation is thatT2D is mediated by loss of production or function of the anti-inflammatory hormone adiponectin. In our previous report, AdipoRon, an adiponectin receptor agonist, and AdipoAI, a newly discovered, more specific agonist, attenuated T2D-associated inflammation by inhibiting osteoclastogenesis and LPS-induced endotoxemia. Autophagy plays an important role during osteoclast differentiation and function. The impact of AdipoAI on osteoclast function and autophagy involved in osteoclastogenesis is not known. Here, we compare AdipoRon and AdipoAI potency, side effects and mechanism of action in T2D-associated periodontitis. METHODS: The RAW 264.7 cell line was used for in vitro studies. We analyzed the potential cytotoxicity of AdipoAI using the CCK-8 assay. The anti-osteoclastogenic potential of AdipoAI was studied by real-time qPCR and tartrate-resistant acid phosphatase staining. The actions of AdipoAI involved in autophagy were tested by real-time qPCR, western blot and immunofluorescence staining. In the diet-induced obesity model of T2D, we investigated the impact of AdipoAI on fasting blood glucose, alveolar bone loss, and gingival inflammation in mice with experimental periodontitis. RESULTS: AdipoRon inhibited osteoclastogenesis and AdipoAI inhibited osteoclastogenesis at lower doses than AdipoRon without any cytotoxicity. In DIO mice with experimental periodontitis, AdipoAI reduced mouse body weight in 14 days, reducing fasting glucose levels, alveolar bone destruction, osteoclast number along the alveolar bone surface, and decreased the expression of pro-inflammatory factors in periodontal tissues. AdipoAI and AdipoRon also enhanced LC3A/B expression when cultured with RANKL.3-Methyladenine, a known autophagy inhibitor, decreased LC3A/B expression and reversed the inhibition of osteoclastogenesis during AdipoAI treatment. CONCLUSIONS: Our results demonstrate that AdipoAI ameliorates the severity of T2D-associated periodontitis by enhancing autophagy in osteoclasts at lower doses than AdipoRon without demonstrable side effects. Thus, AdipoAI has pharmaceutical potential for treating diabetes-associated periodontal disease.

Background Long non-coding RNA (lncRNA) NUTM2A antisense RNA 1 (NUTM2A-AS1) has been reported to be abnormally up-regulated in pulpitis tissues. However, the function of NUTM2A-AS1 in pulpitis remains unclear. The aim of this study was to investigate the role and working mechanism of NUTM2A-AS1 in pulpitis using lipopolysaccharide (LPS)-treated human dental pulp cells (HDPCs). Methods 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, flow cytometry and lactate dehydrogenase (LDH) release detection assay were conducted to analyze the viability of HDPCs. Cell inflammatory response was analyzed through measuring the protein levels of interleukin-6 (IL-6) and IL-8. Western blot assay and quantitative real-time polymerase chain reaction (qRT-PCR) were applied to measure protein expression and RNA expression, respectively. Bioinformatic database StarBase was used to predict the possible targets of NUTM2A-AS1 and let-7c-5p, and dual-luciferase reporter assay was conducted to verify these intermolecular interactions. Results LPS stimulation restrained cell viability and induced cell apoptosis and inflammation of HDPCs. LPS exposure up-regulated the expression of NUTM2A-AS1 and High-Mobility Group Box 1 (HMGB1) and down-regulated the level of let-7c-5p. LPS-induced injury in HDPCs was partly attenuated by the silencing of NUTM2A-AS1 or HMGB1. Let-7c-5p was confirmed as a direct target of NUTM2A-AS1, and let-7c-5p bound to the 3′ untranslated region (3′UTR) of HMGB1 messenger RNA (mRNA) in HDPCs. HMGB1 overexpression largely overturned NUTM2A-AS1 silencing-mediated effects in LPS-induced HDPCs. Conclusion NUTM2A-AS1 knockdown attenuated LPS-induced damage in HDPCs partly through targeting let-7c-5p/HMGB1 axis.

The Intergroup 0116 study has demonstrated a significant survival benefit for completely resected (R0) gastric cancer patients treated with a fluorouracil/leucovorin chemoradiotherapy regimen. However, this regimen is also toxic and less effective in terms of distant disease control. Therefore, a more efficacious and safer regimen is urgently needed. Patients with R0 resected gastric carcinoma received up to two 21-day cycles of postoperative adjuvant preradiation and postradiation DCF chemotherapy (docetaxel 37.5 mg/m2 on days 1 and 8, cisplatin 25 mg/m2 on days 1 to 3, and a continuous infusion of fluorouracil 750 mg/m2 on days 1 to 5), respectively. Chemoradiotherapy between preradiation and postradiation chemotherapy was initiated on day 43 and consisted of intensity-modulated radiotherapy (45 Gy) plus concurrent docetaxel 20 mg/m2 weekly for 5 weeks. A total of 55 patients were evaluated and 76% (42) of patients completed the prescribed therapy. With a median follow-up of 61 months, the 3- and 5-year progression-free survival rates were 67% (95% confidence interval [CI], 54%-80%) and 59% (95% CI, 46%-72%), respectively; and the 3- and 5-year overall survival rates were 72% (95% CI, 60%-84%) and 61% (95% CI, 48%-74%), respectively. The most common grade 3 or greater toxicity, during the chemotherapy phase, was neutropenia (24%). Common grade 3/4 toxicities during concurrent chemoradiotherapy were nausea (32%), vomiting (26%), fatigue (15%), and anorexia (19%). These results demonstrate that this adjuvant regimen is active with an acceptable toxicity profile. A randomized phase 3 trial comparing the Intergroup 0116 chemoradiotherapy regimen with this regimen is underway. This is an open-access article distributed under the terms of the Creative Commons Attribution-Non Commercial-No Derivatives License 4.0 (CCBY-NC-ND), where it is permissible to download and share the work provided it is properly cited. The work cannot be changed in any way or used commercially without permission from the journal. http://creativecommons.org/licenses/by-nc-nd/4.0/

Chronic periodontitis, one of the most prevalent oral diseases, is associated with Porphyromonas gingivalis (Pg) lipopolysaccharide (LPS) infection and has profound effects on type 2 diabetes mellitus (t2DM). Metformin, a well-known antidiabetic agent, has been reported to exert anti-inflammatory effects on various cells. This study aims to investigate the role of metformin on LPS-influenced inflammatory response in human gingival fibroblasts (HGFs).Dose-dependent additive effects of metformin on LPS-influenced HGFs were detected. Cell-counting assay was used to determine effects of metformin and LPS on viability of HGFs. Enzyme-linked immunosorbent assay and quantitative real-time polymerase chain reaction (qRT-PCR) were applied to detect levels of interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α in differently treated cells. Activating transcription factor-3 (ATF3) small interfering (si)RNA transfection was used to determine the mechanism of metformin action, and the transfection efficiency was observed by fluorescence microscope. Effects of ATF3 knockdown were determined by qRT-PCR and Western blot.Results showed that 5 μg/mL Pg LPS and 0.1, 0.5, and 1 mM metformin exhibited no toxicity to HGFs, and metformin inhibited LPS-influenced IL-1β, IL-6, and TNF-α production in a dose-dependent manner. Metformin and LPS could synergistically facilitate ATF3 expression, and ATF3 knockdown abolished inhibitory effects of metformin on LPS-influenced inflammatory cytokine production in HGFs.The present study confirms that metformin suppresses LPS-enhanced IL-6, IL-1β, and TNF-α production in HGFs via increasing ATF3 expression.

Chronic persistent stress is an important cause of gastritis, but the underlying mechanism remains to be further researched, especially the role of the gastric microbiota in this process. Here, we used the water avoidance stress (WAS) test in mouse models for chronic stress‐induced gastritis to investigate the underlying mechanisms of this disease. The effect of stress on the gastric microbiota was analyzed based on 16S rRNA sequencing; the changes in hydrogen sulfide (H2S) and inflammatory cytokine levels in gastric tissues were detected by Western blotting, ELISA, immunofluorescence, and qRT‐PCR. Hematoxylin and eosin staining was used as an indicator of the gastritis histological score. This finding is consistent with previous studies showing that gastric H2S is negatively associated with the inflammatory index and might protect the gastrointestinal tract from inflammation. WAS‐induced gastritis was associated with a reduction in H2S release, which appeared to affect the homeostasis of the gastric microbiota of mice. Inflammation and microbial dysbiosis were partially reversed by sodium hydrosulfide (NaHS) and vitamin B6 (VB6) supplementation, suggesting the therapeutic potential of VB6 supplementation for the treatment of stress‐induced gastritis. Gastritis has a serious impact on health and quality of life. An increasing number of people are suffering from chronic gastritis linked to a high‐stress lifestyle, and our research provides clues for the prevention and treatment of stress‐induced gastritis., Chronic persistent stress is an important cause of gastritis, but the underlying mechanism remains to be further researched. Here, we used the water avoidance stress test in mouse models for chronic stress‐induced gastritis to investigate the underlying mechanisms of this disease. Stress‐induced gastritis was associated with a reduction in hydrogen sulfide release, which appeared to affect the homeostasis of the gastric microbiota of mice.

Mammalian diabetes mellitus which contains altered microenvironment always impairs diverse cellular processes such as osteogenesis, angiogenesis and tissue regeneration via different mechanisms. For researches in materials science, modifying the ability of osteogenesis and angiogenesis in dental implants shows its significant importance. Nano-structure designing is considered as a facile strategy to improve the surface bioactivity of the implants. In this study, the nanorod-structured hydroxyapatite (HA) coatings on Ti-6Al-4V implants were facilely designed by the combination of plasma-spraying and hydrothermal treatment via varying reaction media. Intriguingly, hydrothermal treatment eliminated the glassy phase and impurity phases of HA coatings, and nanorod-structured surface was successfully constructed under hydrothermal treatment in Na₃PO₄ solution. Additionally, the HA coatings with nanorod-structured surface effectively promoted the adhesion and proliferation and further enhanced osteogenic differentiation of DM-rBMSCs in vitro. Moreover, the osseointegration of Ti-6Al-4V implants with nanorod-structured HA coating was also enhanced in diabetes mellitus rabbit model in vivo. Therefore, the nano-structured surface modification of HA coating on Ti-6Al-4V implants could target pathological bone loss via strengthening osteogenesis and angiogenesis and further potentially used as a therapeutic coating to promote diabetic osteointegration.

Purpose Local persistence and relapse of disease in the gross tumor volume (GTV) account for the majority of treatment failures after standard chemoradiation therapy. The primary objective of this phase 1 trial was to define the maximum tolerated dose (MTD) of a hyperfractionated radiation therapy (HFRT) boost to the GTV with concurrent weekly paclitaxel and carboplatin after standard-dose chemoradiation therapy, using image guided intensity modulated radiation therapy techniques. Methods and Materials Eligible patients were given weekly doses of paclitaxel (45 mg/m2) and carboplatin (area under the curve 1.5) for 5 weeks with concurrent radiation therapy (50 Gy), immediately followed by an HFRT boost to the GTV with the same chemotherapy regimen. The boost doses were escalated in increments of 7.2 Gy delivered in 6 twice-daily fractions of 1.2 Gy using a modified Fibonacci design. Once the MTD was established, additional patients were treated at that dose to determine the safety. Results Thirty-one patients fulfilled the inclusion criteria. The incidence of dose-limiting toxicity was 0 of 3, 0 of 3, 0 of 3, 1 of 6 (grade 4 esophagitis), 0 of 3, and 2 of 3 (1 case each of grade 5 esophageal fistula and grade 3 pneumotitis) at 7.2, 14.4, 21.6, 28.8, 36, and 43.2 Gy, respectively, indicating an MTD of 36 Gy. Ten patients treated with this MTD showed no dose-limiting toxicities. The most common acute grade 3 or greater toxicities were esophagitis (26%) and neutropenia (19%). Late toxicity of grade 2 esophageal stricture occurred in 4 patients. The overall response rate was 84% (95% confidence interval, 42%-93%) in the entire cohort. The 1-year local control rate was 100% among those receiving a cumulative dose of the MTD or greater. Conclusions The MTD of the HFRT boost after standard chemoradiation therapy in the setting of concurrent chemotherapy was 36 Gy, resulting in the cumulative tumor dose of 86 Gy in patients primarily with advanced intrathoracic/cervical esophageal squamous cell carcinomas and not adenocarcinomas of the gastroesophageal junction. A phase 2 study to further evaluate this regimen is underway.

Purpose: This article compares the dosimetric differences between jaw tracking and no jaw tracking technique in static intensity-modulated radiation therapy plans of large and small tumors. Methods: Eight plans with large tumor (nasopharyngeal carcinoma, volume range: 510.9 to 768.0 cm3) and 8 plans with small tumor (single brain metastasis, volume range: 5.3 to 9.9 cm3) treated with jaw tracking on Varian EDGE LINAC were chosen and recalculated with no jaw tracking to study the dosimetric differences. We compared the differences of organ-at-risk doses (Dmax, Dmean), monitor units, and γ passing rate of plan verification (3mm/3%, threshold 10%; 2mm/2%, threshold 10%) between the 2 techniques. Results: The organ-at-risk doses of nasopharyngeal carcinoma cases having jaw tracking are all less than those with no jaw tracking. The Dmax and Dmean of organ-at-risks reduced 0.61% to 17.65% and 2.17% to 19.32%, P < .05, respectively. In cases with single brain metastasis, the organ-at-risk doses with jaw tracking were also lower than no jaw tracking. The Dmax and Dmean of organ-at-risk doses reduced 0.84% to 1.52% and 0.90% to 1.86%, P < .05, respectively. The monitor units for the large tumor and small tumor were increased by 2.41% and 1.1%, respectively. The γ passing rates (3mm/3%, th10%; 2mm/2%, th10%) of nasopharyngeal carcinoma plans are 99.89% ± 0.06% (jaw tracking) versus 99.56% ± 0.19% (no jaw tracking; P = .127); 97.15% ± 0.98% (jaw tracking) versus 91.90% ± 1.40% (no jaw tracking; P = .000), and the γ passing rates (3mm/3%, th10%; 2mm/2%, th10%) of brain metastasis plans are 99.97% ± 0.05% (jaw tracking) versus 99.44% ± 1.24% (no jaw tracking; P = .251), 98.65% ± 1.27% (jaw tracking) versus 93.35% ± 2.72% (no jaw tracking; P = .000). Conclusion: Jaw tracking can reduce the dose of organ-at-risks compared to no jaw tracking, and the effect is more significant for plans with large tumor. The γ passing rate of plans with jaw tracking is also higher than the plans with no jaw tracking. Although the monitor units in plans of jaw tracking will increase slightly, it is recommended to use jaw tracking in static intensity-modulated radiation therapy both in large and in small tumors.

Background: Chronic persistent stress is an important cause of gastritis, but its mechanism still needs research. Methods: Here we have used a mouse model and the water avoidance stress (WAS) test as a model for chronic stress-induced gastritis to investigate underlying mechanisms of this disease; The effect of stress on gastric flora was analyzed based on 16sRNA sequencing; the changes of hydrogen sulfide (H2S) and inflammatory cytokines from gastric tissues were detected by western blotting, Elisa, immunofluorescence and qRT-PCR; H&E (Haemotoxylin and Eosin) staining was used for the gastritis histology score. Findings: Consistent with the results of previous studies, gastric H2S was negatively associated with the inflammatory index and may protect the gastrointestinal tract from inflammation. WAS-induced gastritis was associated with a reduction in H2S release, which appeared to affect the homeostasis of the gastric microbiota of mice. Inflammation and microbial dysbiosis were partially reversed by sodium hydrosulfide (NaHS) and vitamin B6 (VB6) supplementation, suggesting that VB6 supplementation may have a therapeutic potential for stress-induced gastritis. Interpretation: The secretory disease gastritis can have a serious effect on health and quality of life, with an increasing number of people suffering from chronic gastritis that is linked to a high-stress lifestyle. Our research provides clues to the prevention and treatment of strss-induced gastritis. Funding: The present research was supported by the National Natural Science Foundation of China (No. 81671978, 81471991, 81772143, 81670486, and 81374101). Declaration of Interest: The authors declare no conflicts of interest. Ethical Approval: All animal experiments were approved by the Animal Care Committee and Ethics Commission of Shandong University (LL-201602047).

Clarithromycin (CLA) is a commonly recommended drug for Helicobacter pylori eradication. However, the prevalence of CLA-resistant H. pylori is increasing. Although point mutations in the 23S rRNA are key factors for CLA resistance, other factors, including efflux pumps and regulation genes, are also involved in the resistance of H. pylori to CLA. Guanosine 3′-diphosphate 5′-triphosphate and guanosine 3′,5′-bispyrophosphate [(p)ppGpp)], which are synthesized by the bifunctional enzyme SpoT in H. pylori , play an important role for some bacteria to adapt to antibiotic pressure. Nevertheless, no related research involving H. pylori has been reported. In addition, transporters have been found to be related to bacterial drug resistance. Therefore, this study investigated the function of SpoT in H. pylori resistance to CLA by examining the shifts in the expression of transporters and explored the role of transporters in the CLA resistance of H. pylori . A Δ spoT strain was constructed in this study, and it was shown that SpoT is involved in H. pylori tolerance of CLA by upregulating the transporters HP0939, HP1017, HP0497, and HP0471. This was assessed using a series of molecular and biochemical experiments and a cDNA microarray. Additionally, the knockout of genes hp0939 , hp0471 , and hp0497 in the resistant strains caused a reduction or loss (the latter in the Δ hp0497 strain) of resistance to CLA. Furthermore, the average expression levels of these four transporters in clinical CLA-resistant strains were considerably higher than those in clinical CLA-sensitive strains. Taken together, our results revealed a novel molecular mechanism of H. pylori adaption to CLA stress.

Porphyromonas gingivalis (P. gingivalis) lipopolysaccharide (LPS) can induce the host immune response in periodontitis patients. Human gingival fibroblasts (GFs) play an important role in regulating the host immune response in periodontitis. However, whether GFs isolated from healthy subjects (HGFs) and inflamed ones (IGFs) can modulate different inflammatory response remains problematic. The aim of this study was to investigate the expression of different inflammatory cytokines between HGFs and IGFs after P. gingivalis LPS stimulation. In this study, hematoxylin and eosin (HE) staining was used to assess the inflammation status of gingiva. HGFs and IGFs were stimulated with 1, 5, and 10 μg/ml P. gingivalis LPS for 6, 12, and 24 h. The amount of inflammatory cytokines, interleukin (IL)-1β, IL-6, IL-8 and tumor necrosis factor (TNF)-α, was determined by enzyme-linked immunosorbent assay (ELISA) and quantitative real-time polymerase chain reaction (qRT-PCR). The results showed that gingiva from periodontitis patients presented epithelial hyperkeratosis and abundant inflamed cells in the connective tissue. HGFs participated in the overproduction of IL-8 and IL-1β in a dose- and time-dependent manner; however, IL-6 and TNF-α just showed a dose-response change when stimulated with LPS after 24 h. In IGFs, IL-6, IL-8, IL-1β, and TNF-α could be induced by lower LPS with shorter time stimulation and the dose-response phenomenon was observed in mRNA levels. In conclusion, the resident IGFs do not exhibit LPS tolerance and play an important role in modulating host immune response, which are critical in the immunopathogenesis of periodontal disease.

// Zhekai Hu 1 , Yu Zhang 1 , Zhiyu Li 2 , Yuedi Yu 3, 4 , Wenyan Kang 1, 5 , Yingnan Han 2 , Xiwen Geng 2 , Shaohua Ge 1, 5 , Yundong Sun 2 1 Shandong Provincial Key Laboratory of Oral Tissue Regeneration, School of Stomatology, Shandong University, Jinan, Shandong 250012, People’s Republic of China 2 Department of Microbiology, Key Laboratory for Experimental Teratology of Chinese Ministry of Education, School of Medicine, Shandong University, Jinan, Shandong 250012, People’s Republic of China 3 Shanghai Southwest Weiyu Middle School, Shanghai 200233, People’s Republic of China 4 Laboratory of Oral Tumor Biology, Shanghai Research Institute of Stomatology Ninth People Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, People’s Republic of China 5 Department of Periodontology, School of Stomatology, Shandong University, Jinan, Shandong 250012, People’s Republic of China Correspondence to: Shaohua Ge, email: shaohuage@sdu.edu.cn Yundong Sun, email: syd@sdu.edu.cn Keywords: Helicobacter pylori, Wnt5a, IL-8, chronic periodontitis, inflammation Received: April 30, 2016 Accepted: August 11, 2016 Published: August 20, 2016 ABSTRACT Helicobacter pylori (H. pylori), a pathogen inducing peptic disease, is recently found to be binding to the progress of periodontitis. Most previous studies are case-controlled, and they investigate the risk of H. pylori infection in disease the development of while few studies evaluate the correlation between H. pylori and periodontal pathogens. Therefore, we investigated the correlation between H. pylori infection with periodontal parameters, periodontal pathogens and inflammation. The results indicated that patients with H. pylori showed significantly higher probing depth and attachment loss than those without ( p < 0.05). Among 28 subgingival plaque samples from 14 patients, the frequencies of Porphyromonas gingivalis, Prevotella intermedia, Fusobacterium nucleatum and Treponema denticola were significantly higher with H. pylori infection than those without H. pylori infection ( p < 0.05). However, the frequency of Aggregatibacter actinomycetemcomitans was lower ( p < 0.05). Furthermore, after human acute monocytic leukemia cell line (THP-1) was stimulated with cagA -positive standard strains ( cagA + H. pylori 26695), the expression of periodontitis-related molecules Wnt5a, interleukin 8 (IL-8), interleukin 6 (IL-6) and interferon gamma (IFN-γ) significantly increased ( p < 0.05). Conversely, the expression of tumor necrosis factor alpha (TNF-α) was almost stable. Meanwhile, cagA + H. pylori promoted significantly higher expression of IL-8 and Wnt5a than isogenic cagA mutants strains ( cagA − H. pylori 26695) did. Taken together, our data suggested that H. pylori might promote the growth of some periodontal pathogens and aggravate the progress of chronic periodontitis.

A new type of water phantom which would be specialised for the absorbed dose measurement in total body irradiation (TBI) treatment is developed. Ten millimetres of thick Plexiglas plates were arranged to form a square cube with 300 mm of edge length. An appropriate sleeve-type piston was installed on the side wall, and a tabular Plexiglas piston was positioned inside the sleeve. By pushing and pulling the piston, the length of the self-made water phantom could be varied to meet the required patients' physical sizes. To compare the international standard water phantom with the length-adjustable and the Plexiglas phantoms, absorbed dose for 6-MV X ray was measured by an ionisation chamber at different depths in three kinds of phantoms. In 70 cases with TBI, midplane doses were metered using the length-adjustable and the Plexiglas phantoms for simulating human dimensions, and dose validation was synchronously carried out. There were no significant statistical differences, p > 0.05, through statistical processing of data from the international standard water phantom and the self-designed one. There were significant statistical differences, p < 0.05, between the two sets of data from the standard and the Plexiglas one. In addition, the absolute difference had a positive correlation with the varied depth of the detector in the Plexiglas phantom. Comparing the data of clinical treatment, the differences were all