[Analysis of time for diagnosis of nontuberculous mycobacterial lung disease and its associated factors in a tuberculosis-designated hospital in Shanghai].

Objective: To investigate the pathogenic characteristics, bacteriological diagnosis time and its associated factors among patients with nontuberculous mycobacterial (NTM) lung disease in a large tuberculosis-designated hospital in Shanghai from 2020 to 2021, in order to improve diagnosis efficiency and formulate precision treatment. Methods: On the basis of the Tuberculosis Database in Shanghai Pulmonary Hospital, NTM patients diagnosed by the Department of Tuberculosis between January 2020 and December 2021 were screened. Demographic, clinical and bacterial information were retrospectively collected. Chi-square test, paired-sample nonparametric test and logistic regression model were used to analyze the factors associated with the diagnosis time of NTM lung disease. Results: A total of 294 patients with bacteriologically confirmed NTM lung disease were included in this study, 147 males and 147 females with a median age of 61(46, 69) years. Of them, 227 (77.2%) patients had comorbidity of bronchiectasis. Species identification results showed that Mycobacterium Avium-Intracellulare Complex was the main pathogen of NTM lung disease (56.1%), followed by Mycobacterium kansasii (19.0%) and Mycobacterium abscessus (15.3%). Species such as Mycobacterium xenopi and Mycobacterium malmoense were rarely identified, accounting for a total proportion of only 3.1%. Positive culture rates for sputum, bronchoalveolar lavage fluid and puncture fluid were 87.4%, 80.3% and 61.5%, respectively. Paired-sample analysis showed that the positive rate of sputum culture was significantly higher than that of smear microscopy (87.1% vs. 48.4%, P<0.01), while no statistical difference was observed between sputum and bronchoalveolar lavage fluid on positive culture rate (78.7% vs. 77.3%, P>0.05). Patients with cough or expectoration were observed with 4.04-fold (95%CI 1.80-9.05) or 2.95-fold (95%CI 1.34-6.52) higher probability of positive sputum culture, compared to those without. Regarding bronchoalveolar lavage fluid, female or patients with bronchiectasis had a 2.82-fold (95%CI 1.16-6.88) or 2.38-fold (95%CI 1.01-5.63) higher probability to achieve a positive culture. The median time to diagnosis of NTM lung disease was 32 (interquartile range: 26-42) days. The results of multivariable analysis showed that patients with symptom of expectoration (aOR=0.48, 95%CI 0.29-0.80) needed a shorter diagnosis time in comparison with patients without expectoration. With Mycobacterium Avium-Intracellulare Complex as a reference, lung disease caused by Mycobacterium abscessus needed shorter diagnosis time (aOR=0.43, 95%CI 0.21-0.88), whereas those caused by rare NTM species were observed to require a longer diagnosis time (aOR=8.31, 95%CI 1.01-68.6). Conclusion: The main pathogen causing NTM lung disease in Shanghai was Mycobacterium Avium-Intracellulare Complex. Sex, clinical symptoms and bronchiectasis had an impact on the positive rate of mycobacterial culture. The majority of patients in study hospital were timely diagnosed. Clinical symptoms and NTM species were associated with the bacteriological diagnosis time of NTM lung disease.

[The application of quantitative analysis of EBV DNA loads in plasma and peripheral blood mononuclear cells in Epstein-Barr Virus infection-related diseases in children].

Objective: To explore the correlation of EBV DNA load in two different types of plasma and peripheral blood mononuclear cells (PBMCs) in children with Epstein-Barr Virus (EBV) infection diseases. Methods: A retrospective evaluated was performed on EBV DNA quantification in plasma and PBMCs by qPCR between April, 2019 and December, 2020. The samples were collected from children of 456 cases with EBV infection and 2 306 healthy cases. In EBV infection group, boys were 253 and girls were 203, aged from 8 days to months to 16 years. In healthy group, boys were 1 267 and girls were 1 039, aged from 8 days to 16 years. Results: Infectious mononucleosis (IM) was the most common disease associated with EBV infection 73.68%(336/456). The detection rate of plasma and PBMCs in EBV infection group was 91.89% (419/456)and 99.34% (453/456)respectively, and was 100%(456/456) in plasma or PBMCs. The detection rate of plasma and PBMCs in healthy group was 1.13%(26/2 306) and 30.01%(715/2 306), respectively. Levels of EBV DNA in plasma and PBMCs in EBV infection group [IM, acute infections, pneumonia, post-transplantation lymphoproliferative disorder (PTLD), hemophagocytic lymphohistiocytosis, tonsillitis and lymphadenitis] was significantly higher than those in healthy group (In plasma, Z=-47.18,-34.41,-33.40,-31.71,-24.38,-20.86 and -20.59,respectively; In PBMCs, Z=-33.17,-16.45,-11.33,-9.45,-5.57,-5.16 and -5.45, respectively; P<0.05). In IM group, EBV DNA load in plasma and PBMCs in remission stage was significantly lower than those in infection stage (Z=-11.45, -8.53;P<0.05). In PTLD group, there was significant difference in EBV DNA load in plasma between infection and remission stage (Z=-4.13, P<0.05), while there was no significant difference in EBV DNA load in PBMCs (Z=-0.817, P>0.05). Conclusions: EBV infection was mainly caused by IM. Combined detection of plasma and PBMCs in EBV DNA is valuable for improving diagnosis ability of EBV infection-related diseases, and the load of EBV DNA could be used as a marker.

Presence of pldA and exoU in mucoid Pseudomonas aeruginosa is associated with high risk of exacerbations in non-cystic fibrosis bronchiectasis patients.

OBJECTIVES: To explore the association between the virulence genes exoU and pldA in isolated mucoid Pseudomonas aeruginosa and the clinical outcomes of patients with non-cystic fibrosis (CF) bronchiectasis. METHODS: A prospective observational cohort study was performed in the Shanghai Pulmonary Hospital from October 2012 to January 2015. We consecutively enrolled all non-CF bronchiectasis patients with mucoid P. aeruginosa isolates obtained from bronchoalveolar lavage fluid or sputum. The exposure variable was the presence of virulence gene, exoU or pldA, in the strains. The primary outcome was exacerbation of bronchiectasis. Logistic regression analysis was performed to evaluate the association between virulence genes and exacerbation. RESULTS: The final analysis included 147 patients (mean (SD) age, 57.86 (11.43) years, 101 female subjects) with median (interquartile range) follow-up of 18 (13-26) months. The following factors were relative to exacerbations: body mass index ≤18.5 kg/m2 (odds ratio (OR) = 5.05; 95% confidence interval (CI), 1.37-18.57), length of stay ≥8 days (OR = 2.65; 95% CI, 1.14-6.19) and positive for either virulence gene (OR = 6.80; 95% CI, 1.47-31.37). The gene-positive group had more exacerbations per year (mean 2.37, SD 2.10, n = 33 vs. mean 0.79, SD 0.83, n = 114) and a higher proportion of patients with exacerbation (31/33, 93.94% vs. 74/114, 64.91%). The proportion of patients being exoU or pldA positive increased as the exacerbation frequency of bronchiectasis increased. CONCLUSIONS: The virulence genes exoU and pldA in mucoid P. aeruginosa are significant risk factors for exacerbations in patients with non-CF bronchiectasis.

miR-483 is down-regulated in gastric cancer and suppresses cell proliferation, invasion and protein O-GlcNAcylation by targeting OGT.

MicroRNAs (miRNAs) are involved in the gastric carcinogenesis and progression. Here, we confirmed that miR-483 was frequently decreased in gastric cancer patients. The expression levels of miR-483 were negatively correlated with tumor stage, node metastasis and stromal invasion. Log-rank tests demonstrated that low expression of miR-483 was strongly correlated with poor overall survival in patients with gastric cancer. Moreover, ectopic expression of miR-483 remarkably suppressed gastric cancer cell proliferation by enhancing cell apoptosis and significantly inhibited the invasion of gastric cancer cells, while low expression of miR-483 exhibited the opposite effect. Bioinformatics analysis revealed that OGT was a potential target of miR-483, and miR-483 inhibited the expression level of OGT mRNA by direct binding to its 3'-untranslated region (3'UTR). Expression of miR-483 was negatively correlated with OGT in gastric cancer tissues. In addition, modulation of miR-483 expression could affect the global cellular protein O-GlcNAcylation in gastric cancer cells. Furthermore, silencing of OGT counteracted the effects of miR-483 repression, while its overexpression reversed tumor inhibitory effects of miR-483. In conclusion, our study revealed that miR-483 functions as a tumor suppressor by inhibiting proliferation, invasion and protein O-GlcNAcylation of gastric cancer via targeting OGT, and that miR-483 may serve as prognostic or therapeutic target for gastric cancer.

[Effect of occupational exposure on job burnout in nurses].

Objective: To investigate the effect of occupational exposure on job burnout in nurses, and to analyze the mediating effect of negative emotion between occupational exposure and job burnout and the regulatory effect of supervisor support on occupational exposure and negative emotion. Methods: From September to December, 2015, simple random sampling was used to select 543 nurses from six tertiary hospitals in Zhejiang Province, China. The questionnaires consisted of occupational exposure risk questionnaire, negative emotion questionnaire, supervisor support questionnaire, and job burnout questionnaire. Results: The total score of occupational exposure risk in nurses was 11.43±7.19; the score of emotional exhaustion was 3.19±1.24, the score of low sense of personal accomplishment was 3.02±1.21, and the score of sense of working indifference was 2.24±1.06. There were significant differences in occupational exposure score between nurses with different sexes (t=2.61, P<0.01) and working years (F=4.49, P<0.01) . There were significant differences in the scores of emotional exhaustion and low sense of personal accomplishment in nurses with different sexes (t=5.25, P<0.001) and working years (t=-3.48, P<0.01) . Occupational exposure had positive effects on negative emotion (ß=0.41, P<0.05) , emotional exhaustion (ß=0.47, P<0.05) , sense of working indifference (ß=0.42, P<0.05) , and low sense of personal accomplishment (ß=0.17, P<0.05) . Negative emotion had a partial mediating effect between occupational exposure and emotional exhaustion (total effect size 30.5%, P<0.05) and between occupational exposure and sense of working indifference (total effect size 37.1%, P<0.05) . Negative emotion had a complete mediating effect between occupational exposure and low sense of personal accomplishment (ß=0.08, P>0.05) . Supervisor support negatively regulate the effects of occupational exposure and negative emotion (F=21.73, P<0.001) . Conclusion: In nurses, occupational exposure has a direct positive effect on job burnout and indirectly influences job burnout via negative emotion. Supervisor support can reduce the negative impact of occupational exposure on negative emotion.

Identification of amino acid residues involved in hemin binding in Porphyromonas gingivalis hemagglutinin 2.

Porphyromonas gingivalis (P. gingivalis) is a major etiological agent in the development and progression of chronic periodontitis. It produces cysteine proteases (gingipains), including a lysine-specific gingipain and two arginine-specific gingipains. Heme binding and uptake are fundamental to the growth and virulence of P. gingivalis. The recombinant hemagglutinin 2 domain (rHA2) of gingipain binds hemin with high affinity. The aim of the present work was to identify the key residues involved in its hemin-binding activity. A functional rHA2 was expressed and bound to hemin-agarose, and then digested with endopeptidases. The peptides bound to hemin-agarose were identified by mass spectrometry and the amino acids were assessed by mutation and peptide binding inhibition analysis. The DHYAVMISK sequence was identified in peptides derived from both Asp-N and Lys-C endopeptidase digestions of rHA2. A monoclonal antibody, mAb QB, was produced and its epitope was associated with the DGFPGDHYAVMISK peptide within the HA2 domain. Hemin was shown to competitively inhibit the immunoreactivity of rHA2 or the peptide to mAb QB. The peptide DHYAVMISK inhibited hemin-binding activity; although, this inhibition was not seen when the peptide contained the H1001E mutation (DEYAVMISK). Based on these results, we propose that residue His1001 is involved in the hemin-binding mechanism of the P. gingivalis rHA2 and the peptide containing this residue, DHYAVMISK, may be an inhibitor of hemin binding.

A systematic review and meta-analysis on the efficacy of low-level laser therapy in the management of complication after mandibular third molar surgery.

The successful reduction of postoperative discomfort is of great significance. This review aims to evaluate the efficacy of low-level laser therapy (LLLT) for the reduction of complication caused by impacted mandibular third molars extraction. An extensive literature search up to October 2013 for randomized controlled trials (RCTs) was performed through CENTRAL, PubMed, Embase, Medline, and CNKI. Six RCTs in which involves 193 participants are included in the meta-analysis. Among them, three RCTs exhibit a moderate risk of bias, while the other three show a high bias risk. Compared with placebo laser/control group, pain is significantly reduced with LLLT on the first day (mean difference [MD] = -2.63, 95% confidence interval [CI] -4.46 to -0.79, P = 0.005). The superiority of LLLT in pain control persists on the second day (MD = -2.34, 95% CI -4.61 to -0.06, P = 0.04) and the third day (MD = -3.40, 95% CI -4.12 to -2.68, P < 0.00001). Moreover, LLLT reduces an average of 4.94 mm (MD = 4.94, 95 % CI 1.53 to 8.34, P = 0.004) of trismus compared with placebo laser irradiation in the first 3 days. On the seventh day, the superiority of LLLT also persists (MD = 3.24, 95% CI 0.37 to 6.12, P = 0.03). In the first 3 days after surgery, extraoral irradiation (MD = -0.69, 95% CI -1.30 to -0.08, P = 0.03) and intraoral combined with extraoral irradiation (MD = -0.65, 95% CI -1.15 to -0.15, P = 0.01) reduced facial swelling significantly. On the seventh day, the intraoral combined with extraoral irradiation group (MD = -0.32, 95% CI -0.59 to -0.06, P = 0.02) still showed benefit in relieving facial swelling. However, because of the heterogeneity of intervention and outcomes assessment and risk of bias of included trials, the efficacy is proved with limited evidence. In the future, well-designed RCTs with larger sample size will be required to provide clearer recommendations.

Fibroblasts regulate variable aggressiveness of syndromic keratocystic and non-syndromic odontogenic tumors.

UNLABELLED: Keratocystic odontogenic tumors (KCOTs) are jaw lesions that can be either sporadic or associated with nevoid basal cell carcinoma syndrome, which typically occurs as multiple, aggressive lesions that can lead to large areas of bone destruction and resorption and cause major impairment and even jaw fracture. To clarify the role of fibroblasts in the aggressivness of syndromic (S-) as compared with non-syndromic (NS-) KCOTs, we assessed fibroblasts derived from 16 S- and NS-KCOTs for differences in cell proliferation, multilineage differentiation potential, alkaline phosphatase activity, and osteoclastogenic potential. S-KCOT fibroblasts had proliferative and osteoclastogenic capacity higher than those from NS-KCOTs, as evidenced by higher numbers of tartrate-resistant acid-phosphatase-positive multinuclear cells, expression of cyclooxygenase 2, and ratio of receptor activator of nuclear factor-kappa B ligand to osteoprotegerin. The osteogenic potential was higher for S- than for NS-KCOT fibroblasts and was associated with lower mRNA expression of runt-related transcription factor 2, collagen type I α1, osteocalcin, and osteopontin as well as reduced alkaline phosphatase activity. These results suggest that the distinct characteristics of fibroblasts in KCOTs are responsible for the greater aggressiveness observed in the syndromic subtype. ABBREVIATIONS: AP, alkaline phosphatase; CK, cytokeratin; COL1A1, collagen type I α1; COX-2, cyclooxygenase-2; GM-CSF, granulocyte-macrophage colony-stimulating factor; IL-1α, interleukin 1α; KCOT, keratocystic odontogenic tumor; NBCCS, nevoid basal cell carcinoma syndrome; NS-KCOT, non-syndrome-associated KCOT; OCN, osteocalcin; OPG, osteoprotegerin; OPN, osteopontin; RANKL, receptor activator of nuclear factor-kappa B ligand; Runx2, runt-related transcription factor 2; S-KCOT, syndrome-associated KCOT; TAF, tumor-associated fibroblast; and TRAP, tartrate-resistant acid phosphatase.

First Report of Leaf Spot of Tea Oil Camellia (Camellia oleifera) Caused by Lasiodiplodia theobromae in China.

Tea oil camellia (Camellia oleifera Abel.), one of the most famous woody oil plants, is distributed and cultivated widely in central and southern China for its strong adaptability. In September 2013, tea oil camellia plants with severe leaf spots were observed in commercial production fields located in Wenchang, Hainan Province. Spots were initially chlorotic, became necrotic and black with a chlorotic halo, developing to cover the entire width of the leaves, and leading to leaf death. Isolations were performed by excising pieces of symptomatic leaves from the lesion margin, surface sterilized with 90% ethanol and 0.6% sodium hypochlorite, and then placed them on potato dextrose agar (PDA). Plates were incubated in a sterile chamber at 26 ± 2°C for 2 days. A fungus was consistently isolated on PDA from all 23 diseased leaf samples. Pure cultures were obtained by monosporic culture technique. After 2 to 3 days of incubation at 26 ± 2°C with a 12-h photoperiod, the fungus initially produced white colonies with dense aerial mycelia, which later turned black (6 to 7 days). The mycelium was fast spreading, branched, and septate. Pycnidia were black, globose, ostiolate, and produced in stroma on the medium surface after 28 days at the same culture conditions as above. Conidia were initially unicellular, subovoid, hyaline, thick-walled with granular content, and 19.8 to 28.9 × 11.5 to 15.7 µm (avg. 25.1 × 13.5 µm). Mature conidia were one-septate and dark brown with longitudinal striations. These observed morphological features suggested that the fungus possessed the same characteristics as previously described for Lasiodiplodia theobromae (Pat.) Griffon & Maubl (syn = Botryodiplodia theobromae) (2). For molecular identification, the ITS1-5.8S-ITS2 region and fragments of the ß-tubulin and elongation factor 1-alpha (EF1-α) genes were sequenced and BLASTn searches done in GenBank. Accession numbers of gene sequences submitted to GenBank were KF811055 for ITS region; KJ639047 for ß-tubulin; and KJ639048 for EF1-α. For all genes used, sequences were 99 to 100% identical to reference isolate CBS164.96 of L. theobromae reported in GenBank (NR_111174, EU673110, and AY640258). Hence, both morphological and molecular characteristics confirmed the fungus as L. theobromae. To confirm fungal pathogenicity, ten 1-year-old healthy plants of C. oleifera were inoculated with the fungus. Mycelial plugs (5 mm) taken from a 7-day-old colony growing on PDA were deposited on wounds with a sterilized knife on leaves and covered with moist cotton. Ten additional control plants were treated similarly but with sterile PDA plugs. Plants were maintained in a moist chamber at 26 ± 2°C for 3 days and then in a greenhouse at 25°C and 40% relative humidity. All the inoculated plants produced typical leaf spot symptoms 3 weeks after inoculation. The fungus was consistently re-isolated from all inoculated plants. Control plants did not show any symptoms. L. theobromae has been reported to cause cankers and dieback in a wide range of hosts and is common in tropical and subtropical regions of the world (1,2), but not previously reported causing disease on C. oleifera. To our knowledge, this is the first report worldwide of leaf spot of C. oleifera caused by L. theobromae. References: (1) S. Mohali et al. For. Pathol. 35:385, 2005. (2) E. Punithalingam. Page 519 in: CMI Descriptions of Pathogenic Fungi and Bacteria. Commonwealth Mycological Institute, Kew, Surrey, UK, 1976.

First Report of Burkholderia andropogonis Causing Bacterial Leaf Spot of Betel Palm in Hainan Province, China.

In May 2009, a severe bacterial disease of arecanut (Areca catechu L.) with an incidence of 100% was observed in a plantation of about 8,400 plants in Wenchang City, Hainan Province, China (19°47.171' N, 110°54.335' E). Symptoms consisted of small circular to elongated brown lesions, ranging from 1 to 105 mm in length and 1 to 21 mm in width, surrounded by yellow halos. White colonies, without fluorescent or diffusible pigments, were consistently recovered on King's B Medium plates from lesions surface-sterilized in 70% ethyl alcohol for 1 min. All isolates were gram-negative and each had a single, polar, sheathed flagellum. Isolates were identified as a Burkholderia sp. based on physiological and biochemical tests: oxidase and catalase positive, negative for arginine dihydrolase, gelatin hydrolysis and starch hydrolysis, and negative for acid production from levan (1,3). Sequences (approx. 1,400 bp each) of the 16S rRNA gene amplified from four isolates using primer pair 27F/1492R (2) (GenBank Accession Nos. JX415481, JX415479, JX415482, and JX415483) shared 99% sequence identity with that of Burkholderia andropogonis strain 6369 (DQ786951). Representative isolates Y11 (China General Microbiological Culture Collection Center No. CGMCC 1.12337), Y30 (CGMCC 1.12338), W15, and W20 were compared with B. andropogonis strain NCPPB No. 1012 and all caused a hypersensitive reaction on leaves of Nicotiana benthamiana. Isolate pathogenicity was tested twice with a total of three replications per isolate. Two young leaves each of 2-year-old arecanut plants were infiltrated with a bacterial suspension of 108 CFU/ml, then covered individually with plastic bags for 48 h, and incubated at 100% relative humidity with 16 h of daylight at 25°C by day and 8 h of darkness at 20°C by night. After 7 days, small water-soaked spots with yellow halos were observed and 60 days after inoculation, lesions developed similar to those caused by B. andropogonis in the field. Koch's postulates were fulfilled by reisolating bacteria from typical lesions on inoculated plants. These bacteria were identical to inoculated strains in colony morphology and sequences of the 16S ribosomal RNA gene. To our knowledge, this is the first report of B. andropogonis infection on betel in Hainan Province, mainland China. This disease was first reported in Taiwan, a province of China. Conditions of high humidity and high temperature support disease outbreaks and infection can result in severe economic losses. In 2012, this disease also appeared on a number of plantations located in other counties. As betel is, economically, the second most important crop in Hainan Province, measures should be required to control this disease, especially in typhoon seasons. References: (1) S. H. Hseu et al. Plant Pathol. Bull. 16:131, 2007. (2) D. J. Lane. In: E. Stackebrandt, et al. Nucleic acid techniques in bacterial systematics. John Wiley & Sons, Chichester, United Kingdom, pp. 115-175, 1991. (3) X. Li and S. H. De Boer. Plant Dis. 89:1132. 2005.

Production and characterization of a monoclonal anti-anti-idiotype antibody against fumonisin B(1).

A monoclonal anti-anti-idiotype antibody (mAb3) against fumonisin B(1) (FmB1) was produced from the hybridoma cell line 7C7F4, which was generated by the fusion of P3/NS-1/1-AG4-1 myeloma cells with spleen cells isolated from a Balb/c mouse that had been immunized with the Fab fragments of affinity-purified anti-idiotype antibodies. The mAb3 belongs to the immunoglobulin M, kappa light chain. A direct competitive enzyme-linked immunosorbent assay (dc-ELISA) and an indirect competitive ELISA (idc-ELISA) were established for antibody characterization and toxin analysis. In an idc-ELISA using FmB1-ovalbumin (OVA) as the coating antigen, the concentrations causing 50% inhibition of binding (IC(50)) of mAb3 to the solid-phase FmB1-OVA by free FmB1, FmB2, and FmB3 were found to be 75, 95, and 450 ng/mL, respectively. In the dc-ELISA, the concentration causing IC(50) of FmB1-horseradish peroxidase to the solid-phase mAb3 by free FmB1 was found to be 233 ng/mL. Analysis of 12 samples naturally contaminated with fumonisins with mAb3-based idc-ELISA and polyclonal-based dc-ELISA showed a good correlation between these two methods with a correlation coefficient of 0.76 at p < 0.02. The linear regression slope was found to be y[polyclonal ELISA] = 0.87x[mAb3 ELISA] - 52 ppb.

[Study on drawing aseptic gas in diluting drugs].

In this study, the gas was drawn from sealed aseptic bottles, the blue flame of an alcohol lamp, and the air of the same treatment room. And the gas was put into aseptic solutions of 10% glucose separately and dripped. Then the samples were taken for bacteriaculture at appointed time-points. Meanwhile, the gases were drawn and put into aseptic solutions of 10% glucose separately. Then deactived penicillines were diluted with the solutions separately. Finally, the penicillines were mixed with 10% glucose and dripped. The samples were taken for bacteria-culture in the same way. The results showed that there was no colony existed in the gas from the sealed aseptic bottles and the flame of the alcohol lamp. However, colonies existed in the samples from the air of the treatment room. It is suggested that sealed aseptic gas should be drawn and kept for use in diluting drugs.