Differential airway resistome and its correlations with clinical characteristics in Haemophilus- or Pseudomonas-predominant microbial subtypes of bronchiectasis.

The prevalence and clinical correlates of antibiotic resistance genes (ARGs) in bronchiectasis are not entirely clear. We aimed to profile the ARGs in sputum from adults with bronchiectasis, and explore the association with airway microbiome and disease severity and subtypes. In this longitudinal study, we prospectively collected 118 sputum samples from stable and exacerbation visits of 82 bronchiectasis patients and 19 healthy subjects. We profiled ARGs with shotgun metagenomic sequencing, and linked these to sputum microbiome and clinical characteristics, followed by validation in an international cohort. We compared ARG profiles in bronchiectasis according to disease severity, blood and sputum inflammatory subtypes. Unsupervised clustering revealed a Pseudomonas predominant subgroup (n = 16), Haemophilus predominant subgroup (n = 48), and balanced microbiome subgroup (N = 54). ARGs of multi-drug resistance were over-dominant in the Pseudomonas-predominant subgroup, while ARGs of beta-lactam resistance were most abundant in the Haemophilus-predominant subgroup. Pseudomonas-predominant subgroup yielded the highest ARG diversity and total abundance, while Haemophilus-predominant subgroup and balanced microbiota subgroup were lowest in ARG diversity and total abundance. PBP-1A, ksgA and emrB (multidrug) were most significantly enriched in Haemophilus-predominant subtype. ARGs generally correlated positively with Bronchiectasis Severity Index, fluoroquinolone use, and modified Reiff score. 68.6% of the ARG-clinical correlations could be validated in an independent international cohort. In conclusion, ARGs are differentially associated with the dominant microbiome and clinical characteristics in bronchiectasis.

The airway microbiome mediates the interaction between environmental exposure and respiratory health in humans.

Exposure to environmental pollution influences respiratory health. The role of the airway microbial ecosystem underlying the interaction of exposure and respiratory health remains unclear. Here, through a province-wide chronic obstructive pulmonary disease surveillance program, we conducted a population-based survey of bacterial (n = 1,651) and fungal (n = 719) taxa and metagenomes (n = 1,128) from induced sputum of 1,651 household members in Guangdong, China. We found that cigarette smoking and higher PM2.5 concentration were associated with lung function impairment through the mediation of bacterial and fungal communities, respectively, and that exposure was associated with an enhanced inter-kingdom microbial interaction resembling the pattern seen in chronic obstructive pulmonary disease. Enrichment of Neisseria was associated with a 2.25-fold increased risk of high respiratory symptom burden, coupled with an elevation in Aspergillus, in association with occupational pollution. We developed an individualized microbiome-based health index, which covaried with exposure, respiratory symptoms and diseases, with potential generalizability to global datasets. Our results may inform environmental risk prevention and guide interventions that harness airway microbiome.

Gut Microbiome Signatures in the Progression of Hepatitis B Virus-Induced Liver Disease.

The gut microbiome is associated with hepatitis B virus (HBV)-induced liver disease, which progresses from chronic hepatitis B, to liver cirrhosis, and eventually to hepatocellular carcinoma. Studies have analyzed the gut microbiome at each stage of HBV-induced liver diseases, but a consensus has not been reached on the microbial signatures across these stages. Here, we conducted by a systematic meta-analysis of 486 fecal samples from publicly available 16S rRNA gene datasets across all disease stages, and validated the results by a gut microbiome characterization on an independent cohort of 15 controls, 23 chronic hepatitis B, 20 liver cirrhosis, and 22 hepatocellular carcinoma patients. The integrative analyses revealed 13 genera consistently altered at each of the disease stages both in public and validation datasets, suggesting highly robust microbiome signatures. Specifically, Colidextribacter and Monoglobus were enriched in healthy controls. An unclassified Lachnospiraceae genus was specifically elevated in chronic hepatitis B, whereas Bilophia was depleted. Prevotella and Oscillibacter were depleted in liver cirrhosis. And Coprococcus and Faecalibacterium were depleted in hepatocellular carcinoma. Classifiers established using these 13 genera showed diagnostic power across all disease stages in a cross-validation between public and validation datasets (AUC = 0.65-0.832). The identified microbial taxonomy serves as non-invasive biomarkers for monitoring the progression of HBV-induced liver disease, and may contribute to microbiome-based therapies.

A transcriptomic analysis based on aberrant methylation levels revealed potential novel therapeutic targets for nasopharyngeal carcinoma.

Background: This study aimed to identify potential novel therapeutic targets for nasopharyngeal carcinoma (NPC) by identifying aberrantly methylated-differentially expressed genes (DEGs) and pathways based on a comprehensive bioinformatics analysis. Methods: Eight gene expression data sets and 2 methylation microarray data sets that included NPC and control groups from the Gene Expression Omnibus were identified. Meta-analyses of the DEGs were performed using the online analysis database "NetworkAnalyst". Aberrantly methylated gene loci were obtained from the GEO2R. Aberrantly methylated DEGs were obtained from Venn diagrams. The enrichment analysis was carried out on the "Metascape" website, and the protein-protein interaction (PPI) network construction, network analysis, and visualization of the analysis results were carried out on the "String" website using "Cytoscape" software. Results: In total, 544 hypomethylation high-expression genes and 164 hypermethylation low-expression genes were obtained. The enrichment and PPI network analyses suggested that several pathways and hub genes with abnormal gene expression accompanied by methylation change, including inositol-trisphosphate 3-kinase B (ITPKB), G protein subunit beta 5 (GNB5), FYN proto-oncogene, Src family tyrosine kinase (FYN), LCK proto-oncogene, Src family tyrosine kinase (LCK), nuclear factor of activated T cells 1 (NFATC1), GNAS complex locus (GNAS), protein kinase C beta (PRKCB), zeta chain of T cell receptor associated protein kinase 70 (ZAP70), lysophosphatidic acid receptor 1 (LPAR1), protein kinase C epsilon (PRKCE), tumor protein p53 (TP53), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), fibronectin 1 (FN1), cyclin D1 (CCND1), vascular endothelial growth factor A (VEGFA), HRas proto-oncogene, GTPase (HRAS), signal transducer and activator of transcription 3 (STAT3), fibroblast growth factor 2 (FGF2), amyloid beta precursor protein (APP), and matrix metallopeptidase 2 (MMP2), may be related to the occurrence of nasopharyngeal carcinoma . Conclusions: The identification of novel and important pathways and hub genes and their roles in the occurrence and development of NPC will guide clinical research and the development of pharmaceutical targets.

[Efficacy evaluation of low-level laser therapy on temporomandibular disorder].

OBJECTIVE: To evaluate effectiveness of low-level laser therapy (LLLT) on temporomandibular joint (TMJ) pain. METHODS: The patients with TMJ pain were randomly assigned laser group (n=21) or control group(n=21), once a day for 6 consecutive days of treatment. TMJ pain and function were measured at baseline, just after treatment course, 1 month and 2 months after the treatment. RESULTS: The changes of visual analogue scale (VAS) were appearing over time in both groups but presented statistically significant differences between groups (P<0.001). VAS of laser group decreased faster than that of control group. The same tendency occurred for painless maximum vertical opening (MVO), left lateral excursion (LLE) and right lateral excursion (RLE), which increased faster in laser group. There were no statistically significant differences between groups and evaluation times for protrusion excursion (PE), but an interaction between group and evaluation times existed and should be explored further. CONCLUSION: LLLT is an appropriate treatment for TMJ pain.

The expression profile of microRNAs in a model of 7,12-dimethyl-benz[a]anthrance-induced oral carcinogenesis in Syrian hamster.

BACKGROUND: Non-coding RNA molecules, such as microRNAs, may play an important role in carcinogenesis. Recent studies have indicated that microRNAs are involved in initiation and progression of various malignancies. However, little work has been done to compare the microRNA expression patterns in oral cancer. In this study, we constructed an animal model of oral squamous cell carcinoma to investigate expression profiles of microRNAs in oral carcinogenesis. METHODS: The animal model of oral squamous cell carcinoma was conducted by tri-weekly (Monday, Wednesday, and Friday) painting with 5% DMBA in acetone. Six Syrian hamsters, including three from the treated group and three from the control group, were used as a training group for microRNA microarray analysis. All microarray data were analyzed by Significance Analysis of Microarrays (SAM) and CLUSTER 3.0 software, and this result was further confirmed by qRT-PCR assay. RESULTS: Seventeen microRNAs were differentially expressed in oral squamous cell carcinoma. Five microRNAs (hsa-miR-21, hsa-miR-200b, hsa-miR-221, hsa-miR-338, and mmu-miR-762) were significantly upregulated and twelve microRNAs (hsa-miR-16, hsa-miR-26a, hsa-miR-29a, hsa-miR-124a, hsa-miR-125b, mmu-miR-126-5p, hsa-miR-143, hsa-miR-145, hsa-miR-148b, hsa-miR-155, hsa-miR-199a, and hsa-miR-203) were down-regulated in cancer tissues. The expression levels of hsa-miR-21 and hsa-miR-16 seen with Stem-loop qRT-PCR were also seen in microarray analysis in all samples. CONCLUSION: Our findings identified specific microRNA expression in oral squamous cell carcinoma and suggested that microRNAs have a role in oral carcinogenesis.

[Effect of estrogen on heat shock protein 70 expression in rat masseter muscle].

OBJECTIVE: To investigate the effect of estrogen on heat shock protein (HSP) 70 expression in rat masseter muscle. METHODS: Sixty twelve-week-old female Sprague-Dawley rats were randomly divided into three groups: Sham surgery group (control group), ovariectomy group (OVX group), ovariectomy with estradiol valerate replacement treatment group (OVX/EV group). Half of the animals were sacrificed at 4 and 8 weeks respectively, then the masseter muscle was removed. Immunohistochemistry (IHC) method was employed to study the HSP70 expression in masseter muscle. RESULTS: Compared to control group and OVX/EV group, the expression of HSP70 was significantly lower at 8 weeks in OVX group (P < 0.05). There were no significantly difference between the HSP70 expression of control group and that of OVX/EV group. CONCLUSION: Estrogen may affect HSP70 expression in rat masseter muscle, and estrogen replacement therapy may prevent HSP70 reduction.

[Roles of matrix metalloproteinases and tissue specific inhibitors of metalloproteinases in dentinogenic ghost cell tumor and ghost cell odontogenic carcinoma].

OBJECTIVE: To investigate the roles of matrix metalloproteinases (MMP) and tissue inhibitors of metalloproteinases (TIMP) in dentinogenic ghost cell tumor (DGCT) and ghost cell odontogenic carcinoma (GCOC). METHODS: The expressions of MMP-2, MMP-9, MMP-14, TIMP-1 and TIMP-2 were examined in 15 DGCT cases and 9 GCOC cases by immunohistochemistry. Their mRNA expression in one DGCT case and one GCOC case were investigated by RT-PCT.MMP-2 and MMP-9 protein activities in the two cases were analyzed by gelatin zymography. RESULTS: MMP-9 and TIMP-1 expressions elevated greatly in GCOC, and there was a significant difference (P < 0.05) in TIMP-1 expression between GCOC and DGCT.Pro-MMP-9, MMP-9 activated form, pro-MMP-2, and MMP-2 activated forms were detected in the GCOC case, while pro-MMP-9 and MMP-9 activated form were very faint in the DGCT case. The mRNA level of MMP-9 elevated obviously in the GCOC case, which was similar to that of TIMP-1. CONCLUSIONS: The elevated expression of MMP-9 and TIMP-1 may influence the behaviour of GCOC.

[Effects of 17beta-estradiol on the intracellular calcium of masticatory muscles myoblast in vitro].

OBJECTIVE: To observe the effects of 17beta-estradiol on the intracellular calcium of masticatory muscles myoblast. METHODS: Myoblasts from maxillofacial skeletal muscle of one week old female Sprague-Dawley rats were cultured. Fluo-4-AM as the Ca2+ indicator and the laser confocal microscope system were used to observe the effects of estrogen on the cytoplasmic Ca2+ concentration in the normal pH condition and the acid condition (pH = 6.7). RESULTS: In the normal pH condition, when 17beta-estradiol (10(-9), 10(-8), 10(-7) mol/L) were added to cells cytoplasmic Ca2+ immediately increased then decreased right away, and in the end came into a new Ca2+ homeostasis in the base line. In the acid condition, 17beta-estradiol (10(-9), 10(-8), 10(-7) mol/L) made the cytoplasmic Ca2+ decreased immediately then came into a new Ca2+ homeostasis under the base line. CONCLUSION: The results suggest that estrogen may maintain the skeletal cytoplasmic Ca2+ concentration in a lower level and reduce the cytoplasmic Ca2+ accumulation to keep the normal functions of masticatory muscles myoblast.

[Regulation of PTHrP in proliferation and differentiation of chondrocytes of condyle in fetal mouse].

OBJECTIVE: To investigate the regulatory mechanism of parathyroid hormone-related protein (PTHrP) in proliferation and differentiation of chondrocytes of condyle in fetal mouse. METHODS: Chondrocytes of condyle in fetal mouse were separated and cultured in vitro, the influence of PTHrP on proliferation and differentiation was observed. RESULTS: After two weeks' culture in 0.01 nmol/L, 0.1 nmol/L, 1 nmol/L, 10 nmol/L human PTHrP, there was significant difference in the number of cartilage nodule formed between experiment group and control group (P<0.05), and there was no significant difference in 0.01 nmol/L group (P>0.05). Alkaline phosphatase (ALP) activity was significantly intensified in experiment group and control group (P<0.05). Meanwhile, it was found that this function of promotion was lessened after anti-PTHR antibody used. CONCLUSION: It can be seen that PTHrP, via its receptor, can promote proliferation and differentiation of chondrocytes of condyle, which resemble its modulation mechanism in epiphyseal growth plate cartilage intramembrane in mandibule.

[The correlation between neck lymphy node metastasis and matrixmetalloproteinase-2 expression at the invasive tumor front of oral squamous cell carcinomas].

OBJECTIVE: To explore the correlation between neck lymph node metastasis and matrix metalloproteinase-2 expression at the invasive tumor front of oral squamous cell carcinomas(OSCC). METHODS: Immunohistochemistry LsAB technique was used to observe the expression of MMP-2 at the invasive tumor front and center of OSCC, and the correlation between the expression of MMP-2 in OSCC and neck lymph node metastasis were respectively analyzed by statistics. RESULTS: The results demonstrated that MMP-2 existed in all 71 cases, which the expression of MMP-2 at the OSCC front was more significant than that of MMP-2 at the OSCC center (P < 0.01), and related to neck lymph node metastasis (P < 0.05). CONCLUSION: The expression of MMP-2 at the OSCC front could be considered as an index of judging the present of neck lymph node metastasis of OSCC.

[Infection of human papillomavirus in oral benign epithelial proliferation in children].

OBJECTIVE: To investigate the presence of HPV infection of oral mucosa proliferative lesions in children and determine the associations of HPV types with oral mucosa lesions in children. METHODS: Immunohistochemical method and in situ hybridization techniques were applied to detect human papillomavirus (HPV) infection in biopsies taken from clinical lesions in oral mucosa of 30 children. RESULTS: The most frequent lesions detected were SCP (66.7%), followed by CA and FEH. The HPV viral antigen was present in 73.3% (22/30) of the oral benign epithelial proliferative lesions in children. A high frequency HPV was found in CA (6/6) and SCP (15/20) by means of IHC. In the ISH positive case, high risk HPV 16/18 was observed in 77.3% (17/22). CONCLUSION: This study demonstrates a high prevalence of HPV infection in children's oral mucosa proliferative lesions, and high-risk HPV16/18 are predominant in children's oral mucosa proliferative lesions.