Detection of SARS-CoV-2 in different human biofluids using the loop-mediated isothermal amplification assay: A prospective diagnostic study in Fortaleza, Brazil.

We adopted the reverse-transcriptase-loop-mediated isothermal amplification (RT-LAMP) to detect severe acute respiratory syndrome coronavirus 2 (SARS-Cov-2) in patient samples. Two primer sets for genes N and Orf1ab were designed to detect SARS-CoV-2, and one primer set was designed to detect the human gene Actin. We collected prospective 138 nasopharyngeal swabs, 70 oropharyngeal swabs, 69 salivae, and 68 mouth saline wash samples from patients suspected to have severe acute respiratory syndrome (SARS) caused by SARS-CoV-2 to test the RT-LAMP in comparison with the gold standard technique reverse-transcription quantitative polymerase chain reaction  (RT-qPCR). The accuracy of diagnosis using both primers, N5 and Orf9, was evaluated. Sensitivity and specificity for diagnosis were 96% (95% confidence interval [CI]: 87-99) and 85% (95% CI: 76-91) in 138 samples, respectively. Accurate diagnosis results were obtained only in nasopharyngeal swabs processed via extraction kit. Accurate and rapid diagnosis could aid coronavirus disease 2019 (COVID-19) pandemic management by identifying, isolating, and treating patients rapidly.

[Interpretation of BRAF gene mutation detection by real-time PCR: a study of 8 644 thyroid fine-needle aspiration specimens].

Objective: To study common problems in BRAF gene mutation detection, and conditions for repetition testing using thyroid fine needle aspiration specimens. Methods: A total of 8 644 cases of thyroid fine-needle aspiration specimens at China-Japan Friendship Hospital were collected between February, 2012 and July, 2018. BRAF gene mutation was detected by real-time PCR. Repeat testing was performed in 237 cases when the results were inconsistent with clinical or cytological diagnosis or when uncertain results were obtained. Results: The final positive rates of BRAF mutation was 22.0% (1 897/8 625). Nineteen cases were excluded due to inadequate DNA samples. The average Ct value of internal quality control was 16.061, and the average Ct value of the positive samples was 19.147. Among 237 repeat tests, 51.4% (19/37) continued to have poor DNA quality and 48.6% (18/37) had adequate DNA resulting in 1 positive case and 17 negative cases. In 40 repetition of initial negative cases, results were unchanged. In initial positive cases, 40.4% (40/99) with a difference of Ct value (between BRAF gene and internal quality control) between 8 to 12 turned negative after repetition, 69.8% (37/53) of these cases with a difference of more than 12 turned negative after repetition. The sensitivity and specificity of BRAF mutation were 83.97% and 96.94%, respectively. Conclusions: Difference between BRAF gene Ct value and internal quality control Ct value is recommended as a reliability index for the test result. Cases with a difference greater than 8 should be subjected to repeat testing.

[Clinicopathologic features of atypical type A thymoma].

Objective: To study the clinicopathologic features, immunophenotype and differential diagnosis of atypical type A thymoma. Methods: Clinicopathologic and follow-up data of three cases of atypical type A thymoma from 2004 to 2016 were reviewed. Immunohistochemical staining was performed. Results: All three patients were male with average age of 59 years. Clinically, the lesions presented as anterior mediastinal masses. Grossly, the tumor ranged in size from 4 to 6 cm in greatest dimension and partially enclosed within fibrous capsule. The cut surface was homogenously fleshy, tan to brown in color, with multinodular and lobulated configurations. Cystic changes and necrosis were seen. Microscopically, the tumor was composed of plump spindle or oval-shaped cells arranged in storiform, microcystic, glandular, rosettes-like and hemangiopericytoma-like histological patterns. The tumor cells showed mild cytological atypia and mitotic activity ranged from 4 to 5/10HPF. Necrosis was present in all tumors. All tumors showed diffuse CK(AE1/AE3), CK19, p63 and vimentin expression. TdT, CK20, CD20, CD5 and CD117 were negative. The proliferative index, as measured with Ki-67, was 15.2% to 26.4%. None of the cases had recurrence or metastases during the follow-up period (9 to 27 months). Conclusions: Atypical type A thymoma presents atypical features including tumor necrosis and increased mitotic activity. However, the implications of such atypical features to the biological behavior of the tumor remain to be determined.

[Expression of PD-1/PD-L1 in lung adenocarcinoma and correlation with epidermal growth factor receptor mutation status].

Objective: To investigate the expression of programmed cell death-1(PD-1) and programmed cell death-ligand 1(PD-L1) in lung adenocarcinoma in correlation with clinical pathological parameters, especially with regard to different epidermal growth factor receptor (EGFR) mutation status. Methods: One hundred and nine cases of lung adenocarcinoma were collected during the period from Aug. 2010 to Jan. 2016, including 51 cases of EGFR wild type and 58 cases of EGFR mutations. Immunohistochemistry was used to detect PD-1/PD-L1 protein expression. Chi-square test was used to analyze the correlation between PD-1 and PD-L1 expression, and in correlation with clinicopathological parameters. All statistical analyses run by SAS 9.1 software. Results: The positive rates of PD-1 and PD-L1 expression were 68.8% (75/109) and 27.5% (30/109), respectively, with significant correlation between the two (P<0.05). PD-1 and PD-L1 expression rates were higher in 51 cases with EGFR wild type status (74.5% and 39.2%) than those in 58 EGFR mutation cases (63.8% and 17.2%); PD-1 expression was significantly associated with age (P<0.05); that of PD-L1 was closely correlated with histological type, tumor size, lymph node metastasis and EGFR status (P<0.05). Conclusions: PD-1 and PD-L1 expression profiles and their correlation with EGFR mutations are different from those with native EGFR. PD-L1 overexpression is closely correlated with larger tumor size and lymph node metastasis, suggesting it is a high-grade marker for lung adenocarcinoma.

Comparative chemometric analyses of geographic origins and compositions of lavandin var. Grosso essential oils by mid infrared spectroscopy and gas chromatography.

Lavandin, a sterile hybrid of Lavandula angustifolia P. Mill. x Lavandula latifolia (L.f.) Medikus (Lamiaceae) is a plant widely cultivated for essential oil production in the South of France. Chemometric treatment by mid-infrared (MID-IR) spectroscopy data was assessed for the differentiation of Grosso Lavandin Essential Oils of Controlled Area (GLEOCA) and results were compared to those obtained by gas chromatography for MID-IR short time technique validation. The quantification of the main 13 hydrocarbons and oxygenated compounds generally controlled by industrial perfumers in GLEOCA samples (n=83) of three geographic origins: "Simiane", "Puimoisson" (with two producers) and "Richerenches" and their classification were successfully obtained by partial least square discriminant analysis (PLS-DA) by comparison with gas chromatography. The best prediction results were obtained using first derivate spectral data in the 1800-700 cm(-1) range. The spectroscopic interpretation of regression vectors showed that each geographic origin was correlated to components of GLEOCA. Chemometric MID-IR spectra treatments allowed us to obtain similar results than those obtained by time consuming analytical techniques such as GC and therefore constitute a robust and help fast method for authentication of GLEOCA and should be extended to other essential oils for authentication of geographic origin.

Epstein-Barr virus (EBV) in Chinese pediatric Hodgkin disease: Hodgkin disease in young children is an EBV-related lymphoma.

BACKGROUND: The Epstein-Barr virus (EBV) is thought to be involved in the pathogenesis of some Hodgkin disease (HD) cases. EBV may be associated particularly with childhood HD, a disease rare in the West compared with developing countries. In this study, a large series of Chinese pediatric HD cases has been examined to determine the age-specific prevalence of EBV. METHODS: Paraffin sections from 104 pediatric and 52 adult Chinese HD cases were examined for EBV-RNA (EBERs) and EBV latent membrane protein-1. RESULTS: Most pediatric cases arose in boys and showed an histology of mixed cellularity. Prominent interfollicular involvement was seen frequently in the childhood cases. EBV was identified in tumor cells in 113 of 156 (72%) HD cases but was more frequent in pediatric cases (93 of 104; 89%) compared with adult cases (20 of 52; 38%) (P < 0.01; chi-square test). EBV was found in 86 out of 91 (95%) cases in children aged 3-10 years and in 7 out of 13 (54%) cases in children aged 11-14 years (P < 0.01; chi-square test). The virus was less frequent in cases in young adults than in old adults, although this trend was not significant (P > 0.05; chi-square test). Pediatric HD was associated with EBV irrespective of histologic subtype. In adults, EBV was associated more frequently with mixed cellularity than with other subtypes. CONCLUSION: To the authors' knowledge, this is to date the largest series of pediatric HD cases studied for EBV. Study findings provided further evidence that HD is etiologically heterogeneous. The authors believe that pediatric HD now should be regarded as a distinctive EBV-related lymphoma.

Epstein--Barr virus gene polymorphisms in Chinese Hodgkin's disease cases and healthy donors: identification of three distinct virus variants.

Epstein--Barr virus (EBV) is associated with several malignancies. Specific EBV gene variants, e.g. the BamHI f configuration, a C-terminal region 30 bp deletion in the latent membrane protein-1 (LMP1) gene (del-LMP) and the loss of an XhoI site in LMP1 (XhoI-loss), are found in Chinese cases of nasopharyngeal carcinoma (NPC), suggesting that EBV sequence variation may be involved in oncogenesis. In order to understand better the epidemiology of these EBV variants, they were studied in virus isolates from EBV-positive Chinese cases of Hodgkin's disease (HD; n=71) and donor throat washings from healthy CHINESE: Sequencing was performed of 15 representative EBV isolates, including the first analysis of the LMP1 promoter in Asian wild-type EBV isolates. The following observations were made. (i) Three EBV LMP1 variants were identified, designated Chinese groups (CG) 1--3. In both EBV-associated HD and in healthy Chinese, CG1-like viruses showing del-LMP1 and XhoI-loss were predominant. (ii) CG1viruses were distinct from European and African variants, suggesting that this profile is useful for epidemiological studies. (iii) Specific patterns of mutations were present in the LMP1 promoter in both CG1 and CG2. (iv) The BamHI f variant was not found in Chinese HD, in contrast to Chinese NPC and European HD. This study confirms that EBV isolates in Chinese HD and other tumours differ from those reported in Western cases. However, this reflects the predominant virus strain present in the healthy Chinese population, suggesting that these are geographically restricted polymorphisms rather than tumour-specific strains.

The immunomodulator AS101 restores T(H1) type of response suppressed by Babesia rodhaini in BALB/c mice.

The immunomodulator AS101 has been previously shown to confer protection upon BALB/c mice infected with the intraerythrocytic parasite Babesia rodhaini (B. rodhaini). The present study focuses on the effect of AS101 administration on the acute phase of babesial infection where T helper cell subset patterns-TH1/TH2-were assessed in heavily infected mice. Secretion of cytokines of the TH1 subset (IL-2, IFN-gamma, IL-12) and of the TH2 subset (IL-10, IL-4) as well as TGF-beta was measured following the administration of AS101 2 weeks before parasite infection. Our results demonstrate that the parasites suppress IL-2 protein and IL-12 mRNA and that AS101 upregulates their secretion: IL-2, 8 u/ml vs 34 u/ml, respectively; IFN-gamma protein, 2370 pg/ml vs 4777 pg/ml, respectively. Conversely, babesial infection results in the upregulation of IL-10 and IL-4 proteins and TGF-beta transcripts, whereas AS101 downregulates their production: IL-10, 1800 pg/ml vs 360 pg/ml, respectively; IL-4, 58.3 pg/ml vs 24.5 pg/ml, respectively. A possible escape mechanism induced by B. rodhaini is suggested, starting with IL-10 inhibition of macrophage activities leading to a suppression of the TH1 response and of IL-2 in particular. It is therefore possible that AS101 may protect infected mice by activating cellular-mediated immunity and concurrently balancing the TH subset responses. It is suggested that AS101 may be effective as an antiparasitic drug.

Delay in the onset of systemic lupus erythematosus following treatment with the immunomodulator AS101: association with IL-10 inhibition and increase in TNF-alpha levels.

It has recently been found that in systemic lupus erythematosus (SLE), a multisystem inflammatory disorder characterized by autoantibody production and decreased cellular immune response, increased spontaneous production of IL-10 occurs. The immunomodulator AS101 (ammonium trichloro(dioxoethylene-0,0')tellurate) was previously shown to significantly decrease IL-10 levels in cancer patients and in murine models. This study shows that AS101 inhibits the development of SLE-related autoimmune pathological manifestations. AS101 decreased the spontaneous IL-10 production by mononuclear cells from SLE patients in vitro. In vivo, systemic injection of AS101 to SCID mice transplanted with mononuclear cells from SLE patients significantly decreased serum human IL-10 levels. There was also a decrease in all serum human Ig isotypes, in anti-dsDNA, and in anti-Sm Igs. In the New Zealand Black/New Zealand White/F1 model, AS101 significantly increased serum TNF-alpha and IFN-gamma while decreasing IL-10 levels; these changes were accompanied by a rapid decrease in anti-dsDNA and anti-ssDNA Igs. More importantly, continuous treatment of New Zealand Black/New Zealand White/F1 mice with AS101 for 6 mo led to the development of proteinuria in 30% of the treated mice compared with 100% in PBS-treated mice (p < 0.001). AS101 treatment reduced the level of immmune complex deposition in the glomeruli, prevented glomerular hypercellularity and mesangial expansion and led to a much smaller mean glomerular volume in treated mice (185 +/- 6 vs 428 +/- 47.103 microm3; p < 0.01). We suggest that treatment with a nontoxic immunomodulator such as AS101, previously used in phase II trials on cancer patients, may be an effective therapeutic approach for controlling SLE.

[Primary anorectal malignant melanoma--report of 7 cases and review of literature].

From 1956 to 1985, 7 patients with primary anorectal malignant melanoma were studied by clinicopathology and observed by light and electron microscopes. The sex ratio of male and female was 2:5. The mean age was 59.4 years. The main presenting symptoms were bloody stool and a palpable mass in the anus. Because these symptoms are non-specific, early diagnosis is very difficult. The disease course ranged from 1.5 to 21.5 months with an average of 6.7 months. At present, the combination treatment is advocated, and radical operation is still the treatment of choice. This tumor has a very poor prognosis. Grossly, the tumor often showed nodular or polypoid shape. Histologically, there were various forms, such as epithelial, spindle and balloon-like cells etc.. Ultrastructurally, melanin corpuscles and vimentin (intermediate filaments) were found in all. In this paper, pathogenesis, morphology and prognosis are discussed.