[Etiological diagnostic value of metagenomic next-generation sequencing in peritoneal dialysis-related peritonitis].

Objective: To explore the etiological diagnostic value of metagenomic next-generation sequencing (mNGS) in peritoneal dialysis (PD)-related peritonitis. Methods: The study was a retrospective cohort study. The clinical data of patients with PD-related peritonitis who were treated and underwent microbial cultivation and mNGS test at the same time from June 2020 to July 2021 in the Affiliated Drum Tower Hospital, Medical School of Nanjing University were analyzed. The positive rate, detection time and consistency between mNGS test and traditional microbial culture were compared. Results: A total of 18 patients with age of (50.4±15.4) years old and median dialysis time of 34.0 (12.4, 62.0) months were enrolled in the study, including 11 males and 7 females. Pathogenic microorganisms were isolated in 17 patients by mNGS test, with a positive rate of 17/18, which was higher than 13/18 of microbial culture, but the difference was not statistically significant (P=0.219). Both mNGS test and microbial culture isolated positive pathogenic bacteria in 12 patients, and mNGS test isolated the same types of pathogenic bacteria as microbial cultivation did in 11 patients. In five patients with negative microbial culture, mNGS test also isolated pathogenic microorganisms, including 3 cases of Staphylococcus epidermidis, 1 case of Mycobacterium tuberculosis and 1 case of Ureaplasma urealyticum. In 1 patient, microbial culture isolated pathogenic bacteria (Escherichia coli) whereas mNGS test did not. The detection time of mNGS was 25.0 (24.0, 27.0) h, which was significantly shorter than 89.0 (72.8, 122.0) h of microbial culture (Z=3.726, P<0.001). Conclusions: mNGS test can improve the detection rate of pathogenic microorganisms in PD-related peritonitis and greatly shorten the detection time, and has good consistency with microbial culture. mNGS may provide a new approach for pathogen identification of PD-related peritonitis, especially refractory peritonitis.

CXCL12/CXCR4 Mediates Orthodontic Root Resorption via Regulating the M1/M2 Ratio.

Mechanical force-induced external root resorption is a major clinical side effect of orthodontic treatment. Recent work has revealed that M1 macrophages play a vital role in promoting orthodontic root resorption (ORR), but the mechanism of how mechanical force stimulation increases the M1/M2 macrophage ratio in periodontal tissue is poorly understood. In the current study, we showed that C-X-C motif chemokine 12 (CXCL12)+ periodontal ligament cells (PDLCs) and C-X-C chemokine receptor type 4 (CXCR4)+ monocytes in the periodontal ligament (PDL) were significantly increased after force application with ongoing root resorption, and these effects were partially rescued after force removal in mice. The expression of CXCL12 in PDLCs was increased by force stimulation in a time- and intensity-dependent manner in vitro. Blockage of the CXCL12/CXCR4 axis using CXCR4 antagonist AMD3100 was sufficient to alleviate ORR and reverse the force-enhanced M1/M2 macrophage ratio. Further mechanism exploration showed that Ly6Chi inflammatory monocytes homed in a CXCL12/CXCR4 axis-dependent manner. The number and proportion of CD11b+ Ly6Chi inflammatory monocytes in cervical lymph nodes were significantly increased by force loading, accompanied by decreased CD11b+ Ly6Chi monocytes in the blood. These changes were blunted by intraperitoneal injection of AMD3100. In addition, blockage of the CXCL12/CXCR4 axis effectively reversed M2 suppression and promoted M1 polarization. Collectively, results indicate that force-induced CXCL12/CXCR4 axis mediates ORR by increasing the M1/M2 ratio in periodontal tissues through attracting Ly6Chi inflammatory monocytes and modulating macrophage polarization. The results also imply that AMD3100 is potentially inhibitory to root resorption.

Down-regulation of hsa_circ_0092125 is related to the occurrence and development of oral squamous cell carcinoma.

Circular RNA plays an important role in regulating tumour development and progression and can serve as a biomarker for cancer. This study was performed to investigate the clinical significance of hsa_circ_0092125 expression in oral squamous cell carcinoma (OSCC). The expression of hsa_circ_0092125 in OSCC tissues and cell lines was determined by reverse transcription-quantitative PCR analysis. The association between hsa_circ_0092125 expression and clinicopathological data was determined by χ2 test. Overall survival (OS) curves were created using Kaplan-Meier survival analysis, and the differences were examined by log-rank test. Moreover, univariate and multivariate Cox analysis were employed to evaluate the risk factors of the OSCC prognosis. The expression of hsa_circ_0092125 was significantly down-regulated in OSCC tissues and cell lines. A low expression of hsa_circ_0092125 was associated with clinicopathological factors in OSCC patients, including tumour size, TNM stage, and lymph node metastasis. Kaplan-Meier survival analysis indicated that the OS time was shorter in OSCC patients with a lower hsa_circ_0092125 expression level than in those with a higher expression level. In addition, univariate and multivariate Cox analysis identified lower hsa_circ_0092125 expression, tumour size, TNM stage, and lymph node metastasis as independent risk factors for the OSCC prognosis. Thus, down-regulated expression of hsa_circ_0092125 might serve as a biomarker of the OSCC prognosis.

The safety of oral fluconazole during the first trimester of pregnancy: a systematic review and meta-analysis.

BACKGROUND: Oral fluconazole is used to treat vulvovaginal candidiasis during pregnancy. However, there are concerns regarding the pregnancy outcomes following exposure to fluconazole. OBJECTIVES: To evaluate the pregnancy outcomes associated with exposure to oral fluconazole during the first trimester of pregnancy. SEARCH STRATEGY: A systematic literature search was conducted to identify relevant studies published from inception until April 2019. SELECTION CRITERIA: Relevant English-language citations using the terms oral fluconazole and pregnancy in humans. DATA COLLECTION: Two reviewers independently abstracted data and assessed study quality. MAIN RESULTS: Oral fluconazole use during the first trimester of pregnancy was marginally associated with an increased risk of congenital malformations (odds ratio [OR] 1.09, 95% CI 0.99-1.2, P = 0.088; n = 6 studies), whereas in the subgroup analysis, this association existed only for high-dose users (>150 mg) (OR 1. 19, 95% CI 1.01-1.4, P = 0.039; n = 2). Exposure to fluconazole also increased the risk of heart malformations (OR 1.31, 95% CI 1.09-1.57, P = 0.003; n = 4), cardiac septal defects (OR 1.3, 95% CI 1.1-1.67, P = 0.047; n = 3), and tetralogy of Fallot (OR 3.39 95% CI 1.71-6.74, P < 0.001; n = 2) in the offspring. In addition, exposure to fluconazole was significantly associated with an increased risk of spontaneous abortion (OR 1.99, 95% CI 1.38-2.88, P < 0.001; n = 3). CONCLUSIONS: Oral fluconazole use during the first trimester of pregnancy appears to be associated with heart malformations and spontaneous abortion, but a causal link cannot be proven. TWEETABLE ABSTRACT: Oral fluconazole during the first trimester of pregnancy may be associated with unfavourable pregnancy outcomes.

Induction of Salivary Gland-Like Cells from Dental Follicle Epithelial Cells.

The dental follicle (DF), most often associated with unerupted teeth, is a condensation of ectomesenchymal cells that surrounds the tooth germ in early stages of tooth development. In the present study, we aim to isolate epithelial stem-like cells from the human DF and explore their potential differentiation into salivary gland (SG) cells. We demonstrated the expression of stem cell-related genes in the epithelial components of human DF tissues, and these epithelial progenitor cells could be isolated and ex vivo expanded in a reproducible manner. The human DF-derived epithelial cells possessed clonogenic and sphere-forming capabilities, as well as expressed a panel of epithelial stem cell-related genes, thus conferring stem cell properties (hDF-EpiSCs). When cultured under in vitro 3-dimensional induction conditions, hDF-EpiSCs were capable to differentiate into SG acinar and duct cells. Furthermore, transplantation of hDF-EpiSC-loaded native de-cellularized rat parotid gland scaffolds into the renal capsule of nude mice led to the differentiation of transplanted hDF-EpiSCs into salivary gland-like cells. These findings suggest that hDF-EpiSCs might be a promising source of epithelial stem cells for the development of stem cell-based therapy or bioengineering SG tissues to repair/regenerate SG dysfunction.

RNA detection based on cyclic amplification of graphene oxide fluorescence.

OBJECTIVE: Graphene oxide (GO) was prepared and used to adsorb single-stranded DNA. Based on fluorescence quenching ability of GO, cyclic signal amplification was performed using polymerase. Thus, two RNA sequences were detected by fluorescence assay. METHODS: Klenow fragment was designed for the amplification of fluorescence signals and RNA was detected by fluorescence assay. After amplification with Klenow fragment, the lowest limit of detection was 1.0 × 10-11 M, and the response of fluorescent intensity was linear within the concentration range of 1.0 × 10-11 M -1.0 × 10-9 M. By the modification of DNA terminals with different fluorescent groups, two different RNA were detected. RESULTS: Different fluorescent dyes were used to modify the terminals of DNA, and two RNA sequences were detected based on fluorescence. There was no need for product separation and purification before detection. CONCLUSIONS: Detection of RNA based on cyclic amplification of GO fluorescence is fast and simple. It can be used for the analysis of specific RNA sequences in cancer cells.

Overexpression of a Populus trichocarpa H+-pyrophosphatase gene PtVP1.1 confers salt tolerance on transgenic poplar.

The Arabidopsis vacuolar H(+)-pyrophosphatase (AVP1) has been well studied and subsequently employed to improve salt and/or drought resistance in herbaceous plants. However, the exact function of H(+)-pyrophosphatase in woody plants still remains unknown. In this work, we cloned a homolog of type I H(+)-pyrophosphatase gene, designated as PtVP1.1, from Populus trichocarpa, and investigated its function in both Arabidopsis and poplar. The deduced translation product PtVP1.1 shares 89.74% identity with AVP1. Semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) and quantitative real-time PCR analyses revealed a ubiquitous expression pattern of PtVP1.1 in various tissues, including roots, stems, leaves and shoot tips. Heterologous expression of PtVP1.1 rescued the retarded-root-growth phenotype of avp1, an Arabidopsis knock out mutant of AVP1, on low carbohydrate medium. Overexpression of PtVP1.1 in poplar (P. davidiana × P. bolleana) led to more vigorous growth of transgenic plants in the presence of 150 mM NaCl. Microsomal membrane vesicles derived from PtVP1.1 transgenic plants exhibited higher H(+)-pyrophosphatase hydrolytic activity than those from wild type (WT). Further studies indicated that the improved salt tolerance was associated with a decreased Na(+) and increased K(+) accumulation in the leaves of transgenic plants. Na(+) efflux and H(+) influx in the roots of transgenic plants were also significantly higher than those in the WT plants. All these results suggest that PtVP1.1 is a functional counterpart of AVP1 and can be genetically engineered for salt tolerance improvement in trees.

Effects of hypoxia on the immunomodulatory properties of human gingiva-derived mesenchymal stem cells.

The environment of bone marrow mesenchymal stem cells (MSCs) is hypoxic, which plays an important role in maintaining their self-renewal potential and undifferentiated state. MSCs have been proven to possess immunomodulatory properties and have been used clinically to treat autoimmune diseases. Here, we tested the effects of hypoxia on the immunomodulatory properties of MSCs and examined its possible underlying mechanisms. We found that hypoxic stimulation promoted the immunomodulatory properties of human gingiva-derived mesenchymal stem cells (hGMSCs) by enhancing the suppressive effects of hGMSCs on peripheral blood mononuclear cells (PBMCs). The proliferation of PBMCs was significantly inhibited, while the apoptosis of PBMCs was increased, which was associated with the Fas ligand (FasL) expression of hGMSCs. The in vivo study showed that systemically infused hGMSCs could enhance skin wound repair, and 24-h hypoxic stimulation significantly promoted the reparative capacity of hGMSCs. For mechanism, hGMSC treatment inhibited the local inflammation of injured skin by suppressing the inflammatory cells, reducing the pro-inflammatory cytokine tumor necrosis factor-α (TNF-α), and increasing anti-inflammatory cytokine interleukin-10 (IL-10), which was promoted by hypoxia. Hypoxia preconditioning may be a good optimizing method to promote the potential of MSCs for the future cell-based therapy.

Immunomodulatory properties of dental tissue-derived mesenchymal stem cells.

In addition to their well-established self-renewal and multipotent differentiation properties, mesenchymal stem cells (MSCs) also possess potent immunomodulatory functions both in vitro and in vivo, which render them a potential novel immunotherapeutic tool for a variety of autoimmune and inflammation-related diseases. The major mechanisms may involve (1) the secretion of an array of soluble factors such as prostaglandin E2 (PGE2 ), indoleamine 2, 3-dioxygenase (IDO), transforming growth factor-ß (TGF-ß), and human leukocyte antigen G5 (HLA-G5); (2) interactions between MSCs and immune cells such as T cells, B cells, macrophages, and dendritic cells. Recently, increasing evidence has supported that MSCs derived from dental tissues are promising alternative sources of multipotent MSCs. We here provide a thorough and extensive review about new findings in the immunomodulatory functions of MSCs derived from several dental tissues, including dental pulp, periodontal ligament, gingiva, exfoliated deciduous teeth, apical papilla, and dental follicle, respectively. The immunomodulatory properties of dental MSCs place them as a more accessible cell source than bone marrow-derived MSCs for cell-based therapy of immune and inflammation-related diseases.

Atmospheric organic nitrogen deposition: analysis of nationwide data and a case study in Northeast China.

The origin of atmospheric dissolved organic nitrogen (DON) deposition is not very clear at present. Across China, the DON deposition was substantially larger than that of world and Europe, and we found significant positive correlation between contribution of DON and the deposition flux with pristine site data lying in outlier, possibly reflecting the acute air quality problems in China. For a case study in Northeast China, we revealed the deposited DON was mainly derived from intensive agricultural activities rather than the natural sources by analyzing the compiled dataset across China and correlating DON flux with NH4(+)-N and NO3(-)-N. Crop pollens and combustion of fossil fuels for heating probably contributed to summer and autumn DON flux respectively. Overall, in Northeast China, DON deposition could exert important roles in agro-ecosystem nutrient management and carbon sequestration of natural ecosystems; nationally, it was suggested to found rational network for monitoring DON deposition.

Comparison of methods for localization of impacted maxillary canines by panoramic radiographs.

OBJECTIVES: The objective of this study was to compare three methods for localization of impacted maxillary canines using only conventional panoramic radiographs. METHODS: The panoramic radiographs of 94 patients (102 impacted maxillary canines) were reviewed and evaluated using the methods magnification, angulation and superimposition. The actual positions of them were decided with cone beam CT images. The predicted positions of impacted canines from the magnification and angulation methods were compared using the McNemar χ(2) test. Sensitivity, specificity, accuracy, positive-likelihood ratio and negative-likelihood ratio were calculated. The canine-incisor index values and α angles of palatally and bucally non-rotated impacted canines were compared using the Mann-Whitney U test. RESULTS: The statistical analysis revealed that there was a significant difference between the magnification and angulation methods (p < 0.01). Using the magnification method, 68.00% of buccal canines and 69.57% of palatal canines could be localized correctly. The results of the angulation method were 28.57% and 84.91%, respectively. The sensitivity of the angulation method for buccal canines was very low. In the superimposition method, 82.98% of the superimposing samples were palatal. CONCLUSIONS: The magnification and angulation methods were not reliable methods for locating the impacted canine with a single panoramic radiograph. Magnification was more successful than the angulation method. Further research is needed on the magnification method. The image superimposition method could be used as an adjunct to others.

Are current definitions of sarcopenia applicable for older Chinese adults?

This study aims to explore whether the current definitions of sarcopenia are applicable to the older Chinese population. Participants were 783 Chinese adults recruited from four regions in Mainland China: Jinan, Guangzhou, Xi'an, and Chengdu. Body composition was measured by dual energy x-ray absorptiometry. Handgrip strength, body weight, and height were measured by trained technicians, and demographic data were collected through questionnaires. Relative appendicular skeletal muscle, skeletal muscle index (SMI=100 × skeletal muscle mass/ body mass) and residuals methods were applied to identify sarcopenia. Compared with young adults, no significant decrease was found in the relative appendicular skeletal muscle (ASM/height2) in older adults. If the criterion of two standard deviations below the mean value of ASM/height2 in young adults is used, none of older adults in this study could be diagnosed with sarcopenia. In addition, compared with the ASM/height2 and residuals methods, SMI shows higher discriminating power in the identifying persons with low handgrip strength. The data suggest that ASM/height2 method may not be appropriate for diagnosis of sarcopenia in Chinese population. However, whether SMI is a better choice remains inconclusive. Prospective studies are needed to clearly define sarcopenia in Chinese population.

Addition of phenolic acids on the reduction of methanol content in wine.

Utilization of phenolic acids, including gallic acid, coumaric acid, caffic acid, cinnamic acid, and ferulic acid, for methanol reduction in wine was investigated. Enzyme activities of pectinesterase and pectin lyase decreased significantly when 0.1 mg/L of gallic acid, coumaric acid, caffic acid, cinnamic acid, or ferulic acid was added. However, no inhibition on polygalacturonase activity was observed when 0.5 mg/L of phenolic acid was added. Methanol content in commercial pectic enzyme (CPE) group increased from 11.53 +/- 1.34 to 56.67 +/- 3.75 ppm in the final products. Adding gallic acid or coumaric acid with CPE inhibited the increase of methanol production. In addition, when 0.2 mg/L of phenolic acid (gallic acid or coumaric acid) was added, the amount of total phenolic acid released from CPE + gallic acid or CPE + coumaric acid groups became higher than CPE group by approximately 466 and 539 mg/L, respectively. In conclusion, the values of lightness, red content, yellow content, total pigment, and total phenolic acid increased in the presence of gallic acid or coumaric acid with CPE, suggesting that adding gallic acid or coumaric acid into winemaking process is a potential method for reducing methanol content, improving wine quality, as well as increasing healthy compounds in wine production.

Determination of random- and blockwise-type de-esterified pectins by capillary zone electrophoresis.

Capillary zone electrophoresis (CZE) was employed to determine the correlations between migration time and degree of esterification (DE) of pectinesterase-de-esterified pectins (PDPs) and alkaline-de-esterified pectins (ADPs) using 50 mM phosphate buffer (pH 6.5) as carrier electrolyte solution and 15 KV as applied voltage. Results showed that pectins with higher DEs exhibited shorter migration times. Linear correlation (r = 0.995) between migration time and DE of ADPs was observed, whereas down-curve correlation in PDPs was observed, regardless of the capillary length used (effective length, 30 and 60 cm). In addition, PDP appeared to migrate faster than ADP with the same DE under the same experimental conditions.

Basic fibroblast growth factor as a growth factor for SRV-2-infected simian retroperitoneal fibromatosis cells, an animal model for AIDS related Kaposi's sarcoma.

Basic fibroblast growth factor (bFGF) and vascular endothelial cell growth factor (VEGF) were demonstrated to be important factors sustaining the growth of Kaposi's sarcoma. RF cells were used to provide a model to study the pathogenesis of Kaposi's sarcoma. In this paper, we demonstrated that bFGF is present in the RF cells, cultured media, and tissues from monkey. The biological activities of bFGF on RF cells were also studied in vitro with serum-free media. The bFGF from serum-free-conditioned media is biologically active to stimulate RF cells in certain media condition. The mitogenic effect was abrogated by sheep neutralizing anti-bFGF antibody. Furthermore, the effect of antibody was reversed by the addition of exogenous bFGF. ELISA measurements indicating the growth potency of conditioned media correlated with the amount of bFGF in the conditioned media. The data from flow cytometry demonstrated the co-existence of SRV-2 and bFGF among RF cells and RF tissues. Immunohistochemical staining of RF tissue blocks for bFGF revealed that bFGF was present in the tumor and the presence of bFGF was not caused by the artifact of tissue culture. These results indicate that bFGF is an important growth factor to promote RF cell growth in vitro and RF tumor in vivo. Further studies are required to determine the relationship between the interaction of bFGF, SRV-2, and VEGF. This model also provides an adequate alternative to the model induced by simian immunodeficiency virus (SIV) to study the Kaposi's sarcoma.

Change in particle size of pectin reacted with pectinesterase isozymes from pea (Pisum sativum L.) sprout.

Four pectinesterase (PE) isozymes were isolated by CM-Sepharose CL-6B chromatography from etiolated pea (Pisum sativum L.) sprouts and then reacted with citrus pectin (degree of esterification = 68%, 30-100 kDa) to observe the change in pectin particle size using a laser particle size analyzer. After incubation of a pectin-PE mixture (pH 6.5) at 30 degrees C for 4 h, PE 1 was observed to catalyze the transacylation reaction most remarkably, increasing the particle size from approximately 50-70 to approximately 250-350 nm, followed by PE 3, PE 2, and PE 4.

Quantification of L-ascorbic acid and total ascorbic acid in fruits and spinach by capillary zone electrophoresis.

A standard curve for the quantification of L-ascorbic acid (L-AA) by capillary zone electrophoresis (CZE) was established, and the quantification of ascorbic acid and total ascorbic acid in fruits (lemon, Sunkist, and pineapple) and spinach were performed using D-isoascorbic acid (D-IAA) as an internal standard. The minimum detection limits (MDLs) for L-AA and D-IAA were determined to be 1 and 2 microg/mL, respectively, at 265 nm. Dehydroascorbic acid (DHAA) in fruits and spinach was quantified in the presence of DL-homocysteine. The recoveries for L-AA in these juices were between 95 and 105%.

[Clinical observation on effect of chemotherapy combined with Chinese medicine in treating advanced tumor patients and on immunologic parameters].

OBJECTIVE: To observe the effects of chemotherapy combined with Chinese herbal medicine (Spleen-Kidney tonifying) in treating advanced tumor patients and on immune parameters. METHODS: One hundred and one advanced tumor patients were randomly divided into the treated group (54 patients) treated by Chinese medicine combined with chemotherapy, and the control group (47 patients) treated by chemotherapy alone. After being treated for 8 weeks (2 treatment courses), the changes of tumor size, body weight, Karnofsky scores, immunologic parameters, peripheral blood cells, as well as the toxic and side-effects were also studied. RESULTS: Improvement of various degree was obtained on the immunologic parameters such as CD3, CD4, CD4/CD8, NK cells, and the quality of life, as Karnofsky score in the treated group, in comparing with those in the control group the difference was significant (P < 0.05, P < 0.01). But on behalf of the toxic and side-effects in advanced patients, there was no significant difference between the two groups. CONCLUSION: Chemotherapy combined with Chinese Spleen-Kidney tonifying drugs could improve the immunologic functions in the advanced tumor patients.

Development of mustard oil-induced hyperalgesia in rats.

Age-dependent changes in nociceptive responses were investigated using either the electromyogram (EMG) recorded from the hamstring muscle in response to electrical stimulation of the hind foot in spinal transected rats or measurement of the tail-flick (TF) reflex latency in intact rats. The development of hyperalgesia produced by topical application of mustard oil was subsequently studied. In experiments involving EMG recordings, rats were tested from day 2 to day 34 after birth (4-day interval) and as adults. In experiments involving measurement of the TF reflex, rats were tested from day 5 to day 30 after birth (5-day interval) and as adults. It was found that the latency and the duration of an early component of the EMG decreased with an increase in animal age, and was similar to adult animals at approximately 18 days after birth. The thermal tail withdraw threshold was lower in pups in comparison with older rats, and took more than 30 postnatal days to become similar to that of adult rats. Although nociceptive behaviors such as biting, body movement, and vocalization could be produced in intact rats by mustard oil in rats as young as 5 days old, the intensities of these responses were subjectively less than those of adult rats. Mustard oil application enhanced significantly the EMG response to electrical stimulation and the effect increased with increasing age. Similarly, mustard oil applied to a hind leg facilitated the TF reflex (decreased response latency). In both experiments, it took approximately 34-40 postnatal days for mustard oil-produced hyperalgesia to become similar to that of adult rats. These data confirm that nociceptive processing is not mature in the young animal and that a developmental period after birth is required for hyperalgesia-related mechanisms to mature.