Periodontitis and osteoporosis: a two-sample Mendelian randomization analysis.

The incidences of periodontitis and osteoporosis are rising worldwide. Observational studies have shown that periodontitis is associated with increased risk of osteoporosis. We performed a Mendelian randomization (MR) study to genetically investigate the causality of periodontitis on osteoporosis. We explored the causal effect of periodontitis on osteoporosis by MR analysis. A total of 9 single nucleotide polymorphisms (SNP) were related to periodontitis. The primary approach in this MR analysis was the inverse variance-weighted (IVW) method. Simple median, weighted median, and penalized weighted median were used to analyze sensitivity. The fixed-effect IVW model and random-effect IVW model showed no significant causal effect of genetically predicted periodontitis on the risk of osteoporosis (OR=1.032; 95%CI: 0.923-1.153; P=0.574; OR=1.032; 95%CI: 0.920-1.158; P=0.588, respectively). Similar results were observed in simple mode (OR=1.031; 95%CI: 0.780-1.361, P=0.835), weighted mode (OR=1.120; 95%CI: 0.944-1.328, P=0.229), simple median (OR=1.003; 95%CI: 0.839-1.197, P=0.977), weighted median (OR=1.078; 95%CI: 0.921-1.262, P=0.346), penalized weight median (OR 1.078; 95%CI: 0.919-1.264, P=0.351), and MR-Egger method (OR=1.360; 95%CI: 0.998-1.853, P=0.092). There was no heterogeneity in the IVW and MR-Egger analyses (Q=7.454, P=0.489 and Q=3.901, P=0.791, respectively). MR-Egger regression revealed no evidence of a pleiotropic influence through genetic variants (intercept: -0.004; P=0.101). The leave-one-out sensitivity analysis indicated no driven influence of any individual SNP on the association between periodontitis and osteoporosis. The Mendelian randomization analysis did not show a significant detrimental effect of periodontitis on the risk of osteoporosis.

Periodontitis and osteoporosis: a two-sample Mendelian randomization analysis

Abstract The incidences of periodontitis and osteoporosis are rising worldwide. Observational studies have shown that periodontitis is associated with increased risk of osteoporosis. We performed a Mendelian randomization (MR) study to genetically investigate the causality of periodontitis on osteoporosis. We explored the causal effect of periodontitis on osteoporosis by MR analysis. A total of 9 single nucleotide polymorphisms (SNP) were related to periodontitis. The primary approach in this MR analysis was the inverse variance-weighted (IVW) method. Simple median, weighted median, and penalized weighted median were used to analyze sensitivity. The fixed-effect IVW model and random-effect IVW model showed no significant causal effect of genetically predicted periodontitis on the risk of osteoporosis (OR=1.032; 95%CI: 0.923-1.153; P=0.574; OR=1.032; 95%CI: 0.920-1.158; P=0.588, respectively). Similar results were observed in simple mode (OR=1.031; 95%CI: 0.780-1.361, P=0.835), weighted mode (OR=1.120; 95%CI: 0.944-1.328, P=0.229), simple median (OR=1.003; 95%CI: 0.839-1.197, P=0.977), weighted median (OR=1.078; 95%CI: 0.921-1.262, P=0.346), penalized weight median (OR 1.078; 95%CI: 0.919-1.264, P=0.351), and MR-Egger method (OR=1.360; 95%CI: 0.998-1.853, P=0.092). There was no heterogeneity in the IVW and MR-Egger analyses (Q=7.454, P=0.489 and Q=3.901, P=0.791, respectively). MR-Egger regression revealed no evidence of a pleiotropic influence through genetic variants (intercept: -0.004; P=0.101). The leave-one-out sensitivity analysis indicated no driven influence of any individual SNP on the association between periodontitis and osteoporosis. The Mendelian randomization analysis did not show a significant detrimental effect of periodontitis on the risk of osteoporosis.

Intricate role of intestinal microbe and metabolite in schizophrenia.

BACKGROUND: The brain-gut axis has gained increasing attention due to its contribution to the etiology of various central nervous system disorders. This study aims to elucidate the hypothesis that schizophrenia is associated with disturbances in intestinal microflora and imbalance in intestinal metabolites. By exploring the intricate relationship between the gut and the brain, with the goal of offering fresh perspectives and valuable insights into the potential contribution of intestinal microbial and metabolites dysbiosis to the etiology of schizophrenia. METHODS: In this study, we used a 16S ribosomal RNA (16S rRNA) gene sequence-based approach and an untargeted liquid chromatography-mass spectrometry-based metabolic profiling approach to measure the gut microbiome and microbial metabolites from 44 healthy controls, 41 acute patients, and 39 remission patients, to evaluate whether microbial dysbiosis and microbial metabolite biomarkers were linked with the severity of schizophrenic symptoms. RESULTS: Here, we identified 20 dominant disturbances in the gut microbial composition of patients compared with healthy controls, with 3 orders, 4 families, 9 genera, and 4 species. Several unique bacterial taxa associated with schizophrenia severity. Compared with healthy controls, 145 unusual microflora metabolites were detected in the acute and remission groups, which were mainly involved in environmental information processing, metabolism, organismal systems, and human diseases in the Kyoto encyclopedia of genes and genomes pathway. The Sankey diagram showed that 4 abnormal intestinal and 4 anomalous intestinal microbial metabolites were associated with psychiatric clinical symptoms. CONCLUSIONS: These findings suggest a possible interactive influence of the gut microbiota and their metabolites on the pathophysiology of schizophrenia.

Analysis of transcriptome expression profiling data in oral leukoplakia and early and late­stage oral squamous cell carcinoma.

The present study screened, potential prognostic biomarkers for oral carcinogenesis. The GSE85195 dataset, which consisted of oral leukoplakia (OL) and early and late-stage oral squamous cell carcinoma (OSCC) samples, was used. The differentially expressed genes (DEGs) in early OSCC vs. OL, late OSCC vs. OL and late OSCC vs. early OSCC groups were screened using the limma package in R. The Short Time-series Expression Miner software package was used to cluster DEGs with similar expression patterns in the course of disease progression (from OL to early and then late-stage OSCC). Moreover, the Database for Annotation, Visualization and Integrated Discovery online analysis tool was used to perform Gene Ontology functional annotation and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis. A protein-protein interaction (PPI) network was also constructed using the Search Tool for the Retrieval of Interacting Genes/Proteins database. Reverse transcription-quantitative PCR was performed to assess the mRNA expression levels of hub node genes in clinical samples, and receiver operating characteristic curve analysis was performed to assess the prognostic value of the hub genes. A total of 4,595, 6,042 and 2,738 DEGs were screened in the early OSCC vs. OL, late OSCC vs. OL and late OSCC vs. early OSCC groups, respectively. A total of 665 overlapping genes were identified when the screened DEGs were compared. Cluster 1 and cluster 7 were identified as the significant clusters, which contained 496 and 341 DEGs, respectively. A PPI network was constructed with 440 interaction pairs. There were five differentially expressed hub nodes identified in different stages from OL to OSCC. The results of the present study indicated that fibronectin 1, signal transducer and activator of transcription 1, collagen type II α1 chain, collagen type X α1 chain and collagen type IV α6 chain might serve as independent diagnostic factors for OL and OSCC, and as prognostic biomarkers for OL carcinogenesis.

Preparation and Characterization of Nanocomposite Hydrogels Based on Self-Assembling Collagen and Cellulose Nanocrystals.

Collagen (Col) hydrogels are an important biomaterial with many applications in the biomedical sector. However, deficiencies, including insufficient mechanical properties and a rapid rate of biodegradation, hamper their application. In this work, nanocomposite hydrogels were prepared by combining a cellulose nanocrystal (CNC) with Col without any chemical modification. The high-pressure, homogenized CNC matrix acts as nuclei in the collagen's self-aggregation process. The obtained CNC/Col hydrogels were characterized in terms of their morphology, mechanical and thermal properties and structure by SEM, rotational rheometer, DSC and FTIR, respectively. Ultraviolet-visible spectroscopy was used to characterize the self-assembling phase behavior of the CNC/Col hydrogels. The results showed an accelerated assembling rate with the increasing loading of CNC. The triple-helix structure of the collagen was preserved with a dosage of CNC of up to 15 wt%. The CNC/Col hydrogels demonstrated an improvement in both the storage modulus and thermal stability which is attributed to the interaction between the CNC and collagen by the hydrogen bonds.

Crosstalk of Exosomal Non-Coding RNAs in The Tumor Microenvironment: Novel Frontiers.

The tumor microenvironment (TME) is defined as a complex and dynamic tissue entity composed of endothelial, stromal, immune cells, and the blood system. The homeostasis and evolution of the TME are governed by intimate interactions among cellular compartments. The malignant behavior of cancer cells, such as infiltrating growth, proliferation, invasion, and metastasis, is predominantly dependent on the bidirectional communication between tumor cells and the TME. And such dialogue mainly involves the transfer of multifunctional regulatory molecules from tumor cells and/or stromal cells within the TME. Interestingly, increasing evidence has confirmed that exosomes carrying regulatory molecules, proteins, and nucleic acids act as an active link in cellular crosstalk in the TME. Notably, extensive studies have identified non-coding RNAs (ncRNAs), including long non-coding RNAs (lncRNAs), microRNAs (miRNAs), and circular RNAs (circRNAs), that could be encapsulated by exosomes, which regulate the coordinated function within the TME and thus participate in cancer development and progression. In this review, we summarize recent literature around the topic of the functions and mechanisms of exosomal ncRNAs in the TME and highlight their clinical significance.

Fabrication and Characterization of Gelatin/Polyvinyl Alcohol Composite Scaffold.

In this study, porous scaffold materials based on polyvinyl alcohol (PVA) and gelatin (Gel) were successfully fabricated and characterized. The mechanism of the reaction, morphology, and crystallinity were investigated by Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), X-ray diffraction (XRD), and X-ray photoelectron spectroscopy (XPS). In addition, thermogravimetric analysis (TGA) was performed together with differential scanning calorimetry (DSC) for examining the thermostability and phase transformation of the scaffolds. Degradation and swelling studies of PVA/Gel composite scaffold materials were performed in phosphate-buffered saline. Finally, the mechanical performances had been determined. According to the results, the polymer matrix that was formed by the combination of PVA and gelatin had better thermal stability. The synthesized composite scaffold was amorphous in nature. The addition of gelatin did not affect the fishbone-like microstructure of PVA, which ensures the excellent mechanical properties of the PVA scaffold. The denaturation temperature and elastic modulus of the PVA scaffold were improved by the gelatin addition, but the physical and chemical properties of the PVA scaffold were weakened when the gelatin content exceeded 10%. In addition, the PVA-10G sample has suitable degradability. Therefore, the PVA/Gel composite scaffold might potentially be applied in the field of tissue engineering that demands high strength.

miR-199a Targeting PNRC1 to Promote Keratinocyte Proliferation and Invasion in Cholesteatoma.

INTRODUCTION: miR-199a has been reported as an oncogene of various cancers. However, the biological function and regulatory mechanism of miR-199a in keratinocytes of cholesteatoma are still unclear. METHODS: Detection by qRT-PCR was conducted on miR-199a's expression in both thirty pairs of cholesteatoma tissues and normal skins. For characterizing the function of miR-199a, this research adopted transwell assay, wound healing assay, and CCK8 assays. Under the support of qRT-PCR, efforts were made to investigate the relative expression of candidate target genes. Moreover, the evaluation of the targeting relationship between miR-199a and the candidate target gene was conducted with the dual-luciferase reporter assay. RESULTS: The upregulation of miR-199a was found in cholesteatoma tissues, which facilitated the proliferation, migration, and invasion of HaCaT cells, while its downregulation caused opposite results. CONCLUSIONS: The findings of the present research offer more insights into the molecular mechanism of cholesteatoma progression.

Association Between Abundance of Haemophilus in the Gut Microbiota and Negative Symptoms of Schizophrenia.

Increasing evidence indicates an interaction between dysbiosis of the microbiota and the pathogenesis of schizophrenia. However, limited information is available on the specific microbial communities associated with symptoms of schizophrenia. Therefore, this study aimed to investigate gut microbiota dysbiosis and its relationship with psychopathologies in schizophrenia. We recruited 126 participants and divided them into three groups according to the Diagnostic and Statistical Manual of Mental Disorders, Fifth Edition, criteria-acute group (patients with acute schizophrenia), remission group (patients with schizophrenia in remission), and control group (healthy controls). Psychotic symptoms were evaluated using the Positive and Negative Syndrome Scale. Microbiota compositions, diversity and community structure were evaluated using 16S rRNA sequencing. Pearson's correlation analysis was used to evaluate the association between bacterial taxa and psychotic symptoms. The beta-diversity of microbiota composition in the acute group was distinct from that in the remission and control groups (PC1 = 21.11% vs. PC2 = 12.86%, P = 0.021). Furthermore, Pearson's correlation analysis revealed that abundance of Haemophilus was positively correlated with negative psychiatric symptoms (r = 0.303, P = 0.021), while abundance of Coprococcus was negatively correlated with negative psychiatric symptoms (r = -0.285, P = 0.025). Moreover, abundance of Haemophilus was positively correlated with cognition (r = 0.428, P = 0.009), excitement (r = 0.266, P = 0.037), and depression (r = 0.295, P = 0.020). The study findings suggest that alterations in certain gut microbiota may interfere with psychological symptoms in schizophrenia. Our results provide evidence that may help in the development of therapeutic strategies using microbial-based targets. The data that support the findings of this study have been deposited in the NCBI (https://submit.ncbi.nlm.nih.gov/) with accession number SUB9453991.

Robust Hydrogel Adhesive with Dual Hydrogen Bond Networks.

Hydrogel adhesives are attractive for applications in intelligent soft materials and tissue engineering, but conventional hydrogels usually have poor adhesion. In this study, we designed a strategy to synthesize a novel adhesive with a thin hydrogel adhesive layer integrated on a tough substrate hydrogel. The adhesive layer with positive charges of ammonium groups on the polymer backbones strongly bonds to a wide range of nonporous materials' surfaces. The substrate layer with a dual hydrogen bond system consists of (i) weak hydrogen bonds between N,N-dimethyl acrylamide (DMAA) and acrylic acid (AAc) units and (ii) strong multiple hydrogen bonds between 2-ureido-4[1H]-pyrimidinone (UPy) units. The dual hydrogen-bond network endowed the hydrogel adhesives with unique mechanical properties, e.g., toughness, highly stretchability, and insensitivity to notches. The hydrogel adhesion to four types of materials like glass, 316L stainless steel, aluminum, Al2O3 ceramic, and two biological tissues including pig skin and pig kidney was investigated. The hydrogel bonds strongly to dry solid surfaces and wet tissue, which is promising for biomedical applications.

Polyethyleneimine-functionalized Fe3O4/attapulgite particles for hydrophilic interaction-based magnetic dispersive solid-phase extraction of fluoroquinolones in chicken muscle.

Fluoroquinolone (FQ) residues in foods of animal origin may threaten public health but are challenging to determine because of their low contents and complex matrices. In this study, novel polyethyleneimine-functionalized Fe3O4/attapulgite magnetic particles were prepared by a simple co-mixing method and applied as hydrophilic sorbents for the magnetic dispersive solid-phase extraction (MSPE) of three FQs, i.e., ciprofloxacin, norfloxacin, and enrofloxacin, from chicken muscle samples. The preparation of the magnetic particles was of high reproducibility and the products could be reused many times with high adsorption capacity. The key experimental factors possibly influencing the extraction efficiencies, including sample solution, extraction time, sample loading volume, desorption solution, desorption time, and elution volume were investigated. Under optimum MSPE conditions, the analytes in chicken muscle samples were extracted and then determined by RPLC-MS/MS in MRM mode. Good linearity was obtained for the analytes with correlation coefficients ranged from 0.9975 to 0.9995. The limits of detection were in the range of 0.02-0.08 µg kg-1, and the recoveries of the spiked FQs in chicken muscle samples ranged from 83.9 to 98.7% with relative standard deviations of 1.3-6.8% (n = 3). Compared with the traditional MSPE methods based on hydrophobic mechanism, this hydrophilic interaction-based method significantly simplifies the sample pretreatment procedure and improves repeatability. This method is promising for accurate monitoring of FQs in foods of animal origin.

Ultrasonic assisted microwave synthesis of poly (Chitosan-co-gelatin)/polyvinyl pyrrolidone IPN hydrogel.

The hydrogels based on gelatin cross-linked with chitosan (CS) and polyvinyl pyrrolidone (PVP) were synthesized using microwave and ultrasonic coupling technique in this study. This interpenetrating polymer network (IPN) hydrogels were cross-linked by glutaraldehyde and 1,2-Epoxy-4-vinylcyclohexane. The presence of function groups in the structure of hydrogel films were confirmed using Fourier transform infrared spectroscopy (FT-IR), thermal stability was measured by DSC, and the swelling behaviors were measured gravimetrically in distilled water at the temperature of 27°C. At last, the mechanical properties were tested. The results showed that the hydrogel prepared with microwave and ultrasonic exhibited the highest tensile strength (86.68MPa), comparing with the hydrogel prepared with traditional method and only microwave reactive field. The FT-IR and XRD results showed that the chemical reactions occurred between the NH2 of chitosan and the COOH of gelatin, and the introduction of ultrasound can improve the reaction rate. The hydrogel film gained in microwave and ultrasonic coupling field has the best combination properties. Therefore, the new microwave-ultrasonic coupling technique is the potential technology to prepare the new hydrogel due to less synthesis time.

Multimetallic synergic sedation of a labile sodium atrane: synthesis and characterization of a tetranuclear sodium atrane cation complex.

A series of sodium and aluminum atrane complexes of Na(3)L(THF)(5) (1), [AlLMe][Na(4)L(THF)(6)] (2), AlL(THF) (3), AlNaLMe(THF)(2) (4), and AlNaLOBn(THF)(2) (5), wherein L = tris(2-oxy-4,6-di-tert-butyl-benzyl)amine, were synthesized and characterized by NMR, X-ray crystallography, and elemental analysis. The trinuclear sodium atrane complex of Na(3)L(THF)(5) (1) is labile at room temperature; however, the tetranuclear sodium atrane cation in complex 2 can be stabilized by a multimetallic synergetic effect due to a firm interaction ring of -[Na-O-benzene](3)-. Complex 2 is also the first example of a sodatrane and alumatrane ion-paired complex in which both the cationic and anionic moieties contain an atrane ligand.

Acetonitrile-triaqua-[3-eth-oxy-1,8-(3,6,9-trioxaundecane-1,11-diyldi-oxy)-9H-xanthen-9-one]terbium(III) tris-(perchlorate).

In the title compound, [Tb(CH(3)CN)(C(23)H(26)O(8))(H(2)O)(3)](ClO(4))(3), the Tb(3+) atom is eight-coordinated by one N atom of an acetonitrile molecule, three water O atoms and four ligand O atoms. The Tb(3+) atom is located on one side of the macrocycle and the carbonyl oxygen coordinated to the terbium [Tb1-O1= 2.210 (3) Å] is bent out of the xanthone plane by 0.514 (3) Å. The geometry around terbium is a distorted two-capped trigonal prism.

Rapid detection of intestinal pathogens in fecal samples by an improved reverse dot blot method.

AIM: To develop a new, rapid and accurate reverse dot blot (RDB) method for the detection of intestinal pathogens in fecal samples. METHODS: The 12 intestinal pathogens tested were Salmonella spp., Brucella spp., Escherichia coli O157:H7, Clostridium botulinum, Bacillus cereus, Clostridium perfringens, Vibrio parahaemolyticus, Shigella spp., Yersinia enterocolitica, Vibrio cholerae, Listeria monocytogenes and Staphylococcus aureus. The two universal primers were designed to amplify two variable regions of bacterial 16S and 23S rDNA genes from all of the 12 bacterial species tested. Five hundred and forty fecal samples from the diarrhea patients were detected using the improved RDB assay. RESULTS: The methods could identify the 12 intestinal pathogens specifically, and the detection limit was as low as 103 CFUs. The consistent detection rate of the improved RDB assay compared with the traditional culture method was up to 88.75%. CONCLUSION: The hybridization results indicated that the improved RDB assay developed was a reliable method for the detection of intestinal pathogen in fecal samples.

Bis[2-(1,3-benzothia-zol-2-ylsulfan-yl)eth-yl] ether.

The complete molecule of title compound, C(18)H(16)N(2)OS(4), is generated by crystallographic twofold symmetry, with the O atom lying on the rotation axis. The dihedral angle between the ring systems is 80.91 (2)°. In the crystal, adjacent mol-ecules are connected through π-π stacking inter-actions [centroid-centroid distance = 3.882 (2) Å], forming a three-dimensional network.

(E)-N-[2-(3,5-Di-tert-butyl-2-hydroxy-benzyl-ideneamino)cyclo-hexyl]-4-methyl-benzene-sulfonamide.

In the crystal structure of the title compound, C(28)H(40)N(2)O(3)S, there are two mol-ecules per asymmetric unit; in each of these mol-ecules, the cyclo-hexyl rings adopt chair conformations. The dihedral angles between the benzene rings are 16.89 (9) and 34.11 (9)°. Each mol-ecule contains an intra-molecular O-H⋯N hydrogen bond, and inter-molecular N-H⋯O hydrogen bonds are also present. In both mol-ecules, the methyl groups of one tert-butyl group are disordered over two positions; the site-occupancy factors in both cases are ca 0.6 and 0.4.

[Detection and identification of seven clinical common pathogenic bacteria by oligonucleotide microarray].

OBJECTIVE: Using 16S rDNA and 23S rDNA genes as the target sequences to develop a system based on oligonucleotide microarray and to detect the seven clinical pathogenic bacteria, commonly seen. METHODS: Double polymerase chain reaction (PCR) was applied to amplify the segments of 16S rDNA and 23S rDNA genes of the target bacteria. An oligonucleotide microarray was constructed to simultaneously detect EHEC O157:H7, Vibrio parahaemolyticus, Salmonella sp., Vibrio cholerae, Listeria monocytogenes, Campylobacter jejuni and Shigella sp. Specificity, sensitivity and reproducibility of the microarray during detection were checked. And then microarray was used to detect the microbes in stool specimens of 81 patients with diarrhea and vomiting. RESULTS: The double PCR method could simultaneously amplify the target sequences of 16S rDNA and 23S rDNA genes of the seven pathogens. The sensitivity of the developed oligonucleotide microarray could reach 10(3) cfu/ml but no positive results were presented for non-targeted bacteria. The coefficients of differentiation in one lot or among different lots of the microarray slices were 3.89% - 5.81%. The positive detection rate of the stool specimens by oligonucleotide microarray was 39.5% (32/81), with a coincidence of 96.3% (78/81) for the patients and another coincidence of 96.8% (31/32) for bacterial genus or species identification, when comparing to the results by routine bacteriological examinations. CONCLUSION: The established assay in this study based on oligonucleotide microarray to detect the seven pathogenic bacteria has many advantages such as convenient, rapid, accurate, stable and high flux, which is suitable for clinical specimen examination and epidemiological field investigation.

Selective colorimetric and fluorescent detection of HSO4- with sodium(I), magnesium(II) and aluminium(III) xanthone-crown ether complexes.

Xanthone-crown ether (1) reacts with NaClO(4), Mg(ClO(4))(2) and Al(ClO(4))(3) forming the one-dimensional chain dinuclear polymer [Na(2).1.(ClO(4))(2)] (2), the mononuclear complex [Mg.1.(H(2)O)(2)](ClO(4))(2) (3) and an interesting sandwich complex [Al.(1)(2).(H(2)O)(6)](ClO(4))(3) (4) with different ratios of metal-to-ligand, respectively. The anion recognition experiment results show that the magnesium complex (3) is a good colorimetric and fluorescent detector for HSO(4)(-) with high sensitivity and selectivity.

[Detection and identification of human metapneumovirus by real time reverse transcription PCR].

OBJECTIVE: To develop a rapid, sensitive and specific real time reverse transcription PCR for detecting and identifying human metapneumovirus. METHODS: The Hmpv-L gene of human metapneumovirus was chosen as target gene, the primers and TaqMan probe were designed, and the PCR reaction was optimized systematically. The total RNA was extracted from respiratory specimens, and reverse transcription was performed through random primer. The cDNA was detected by using real time PCR. The specificity, sensitivity and reproducibility of real time PCR were estimated. The real time PCR was applied to detect 180 clinical respiratory specimens. RESULTS: The human metapneumovirus can be detected using real time reverse transcription PCR accurately and quickly, and the sensitivity was 1 copy/microl. The coefficient of variation of intra-assay and inter-assay was less than 5%. Among those 180 specimens, 28 (15.56%) were positive for human metapneumovirus, the clinical diagnoses for these 28 patients were pneumonia (15.60%, 17/109) and bronchiolitis (15.49%, 11/71). 21 positive specimens were from patients under 2 years of age, and 6 positive specimens were from patients between 2 and 5 years of age, only 1 positive specimens was from patients over 5 years. CONCLUSION: It is demonstrated that real time reverse transcription PCR is a reliable, accurate and feasible assay for human metapneumovirus, which has become one of the most important pathogens induced acute respiratory infections in pediatric patients.