Comparative genome-wide identification and characterization of SET domain-containing and JmjC domain-containing proteins in piroplasms.

BACKGROUND: SET domain-containing histone lysine methyltransferases (HKMTs) and JmjC domain-containing histone demethylases (JHDMs) are essential for maintaining dynamic changes in histone methylation across parasite development and infection. However, information on the HKMTs and JHDMs in human pathogenic piroplasms, such as Babesia duncani and Babesia microti, and in veterinary important pathogens, including Babesia bigemina, Babesia bovis, Theileria annulata and Theileria parva, is limited. RESULTS: A total of 38 putative KMTs and eight JHDMs were identified using a comparative genomics approach. Phylogenetic analysis revealed that the putative KMTs can be divided into eight subgroups, while the JHDMs belong to the JARID subfamily, except for BdJmjC1 (BdWA1_000016) and TpJmjC1 (Tp Muguga_02g00471) which cluster with JmjC domain only subfamily members. The motifs of SET and JmjC domains are highly conserved among piroplasm species. Interspecies collinearity analysis provided insight into the evolutionary duplication events of some SET domain and JmjC domain gene families. Moreover, relative gene expression analysis by RT‒qPCR demonstrated that the putative KMT and JHDM gene families were differentially expressed in different intraerythrocytic developmental stages of B. duncani, suggesting their role in Apicomplexa parasite development. CONCLUSIONS: Our study provides a theoretical foundation and guidance for understanding the basic characteristics of several important piroplasm KMT and JHDM families and their biological roles in parasite differentiation.

Establishment of Serum TSH Reference Intervals by an Indirect Method for Healthy Population in Southwest China.

BACKGROUND: Serum thyroid stimulating hormone (TSH) detection is of great clinical significance in monitoring thyroid function. The aim of the present study was to establish reference intervals (RIs) for serum TSH in healthy Han population in Southwest China using data from routine health check-up individuals. METHODS: Healthy subjects (n = 7,116) were enrolled from January 2019 to September 2020. Serum TSH values were determined in the Beckman Coulter UniCel™ DxI 800 Access® immunoassay system. Outliers were identified and removed using Dixon's interactive principle. The 95th percentile range was calculated as RIs for serum TSH. All the statistical analyses were run on R statistical software version 4.0.3. RESULTS: A total of 6,668 (1,324 female and 5,344 male) suitable individuals were included in this study. Serum TSH results showed a non-Gaussian distribution by Kolmogorov-Smirnov test. According to Mann-Whitney U analysis, the serum TSH values for the female group differ from the male group's (p < 0.05). Besides, Spearman's rank correlation test disclosed that no obvious correlation was observed between serum TSH levels and age (r = -0.0039, p > 0.05). Accordingly, the RIs for serum TSH in Southwest China Han population were 0.64 (95% CI: 0.62 to 0.65) to 4.05 (95% CI: 4.02 to 4.09) mIU/L in male and 0.72 (95% CI: 0.67 to 0.77) to 5.66 (95% CI: 5.58 to 5.75) mIU/L in female, respectively. CONCLUSIONS: In this study, the laboratory specific RIs for serum TSH were successfully established by indirect method using the data from health check-up population. It implies that the indirect method is an easy and lowcost pathway for each laboratory to establish its own RIs.

Comparison of the diagnostic performance of fluorescence in situ hybridization (FISH), nuclear matrix protein 22 (NMP22), and their combination model in bladder carcinoma detection: a systematic review and meta-analysis.

BACKGROUND: Emerging studies reported that combination of fluorescence in situ hybridization (FISH) and nuclear matrix protein 22 (NMP22) could increase the sensitivity and specificity of bladder carcinoma (BC) management. Nevertheless, the reports remain inconsistent. This meta-analysis was undertaken to evaluate the diagnostic performance of FISH, NMP22, and their combination model in BC. MATERIALS AND METHODS: A systematic literature search was carried out in PubMed, Embase, Cochrane Library, Web of Science, Chinese National Knowledge Infrastructure, and Wanfang database dated up to October 2018. Suitable studies were identified and raw data were extracted. Meta-analysis was conducted to calculate the global sensitivities, specificities, likelihood ratio, diagnostic odds ratio (DOR), and the areas under the summary receiver operating characteristic (SROC) curves for FISH, NMP22, and their combination model, separately. All the meta-analysis estimates were derived using STATA (version 12.0) and MetaDisc (version 1.4) software packages. RESULTS: Seven eligible studies were included for analysis. The global sensitivities with 95% CI for FISH, NMP22, and their combination model were 0.79 (95% CI: 0.75-0.83), 0.76 (95% CI: 0.71-0.81), and 0.82 (95% CI: 0.75-0.88); specificities were 0.85 (95% CI: 0.76-0.91), 0.70 (95% CI: 0.55-0.81), and 0.90 (95% CI: 0.70-0.97); DORs were 22.215 (95% CI: 10.695-46.144), 7.365 (95% CI: 3.986-13.610), and 41.940 (95% CI: 13.546-129.853); and the areas under the SROC curves were 0.86 (95% CI: 0.82-0.88), 0.79 (95% CI: 0.76-0.83), and 0.90 (95% CI: 0.87-0.92). CONCLUSION: Our systematic review implied that the diagnostic performance of combination model of FISH plus NMP22 may outperform FISH or NMP22 alone in BC detection.

Diagnostic Value of MicroRNA-18a for Gastric Cancer: a Meta-Analysis.

BACKGROUND: Emerging data demonstrated that circulating microRNA-18a (miR-18a) expression level was significantly different between gastric carcinoma individuals and healthy groups, implying that miR-18a may be a potential biomarker for gastric cancer. Nevertheless, the reports remain inconsistent. This meta-analysis was performed to evaluate the diagnostic value of miR-18a in gastric tumor detection. METHODS: All the relevant papers were searched and collected until July 2017 in PubMed, Cochrane Library, Wiley Online Library, Chinese National Knowledge Infrastructure (CNKI), Technology of Chongqing (VIP), and Wanfang Database. Data was extracted from eligible studies. Diagnostic performance of miR-18a for gastric cancer were evaluated using STATA (version 12.0) and MetaDisc (version 1.4) statistical software. RESULTS: Three studies were included in this meta-analysis with a total of 235 gastric cancer patients and 136 controls enrolled. The pooled sensitivity and specificity, positive likelihood ratio (PLR), negative likelihood ratio (NLR), and diagnostic odds ratio (DOR) of circulating miR-18a to discriminate GC patients were 0.76 (95% confidence interval CI: 0.70, 0.81), 0.73 (95% CI: 0.65, 0.80), 2.76 (95% CI: 2.08, 3.65), 0.32 (95% CI: 0.19, 0.55), and 9.12 (95% CI: 4.36, 19.09), respectively. The area under the summary receiver operator characteristic (SROC) curve was 0.82. CONCLUSIONS: miR-18a could be a promising noninvasive biomarker in gastric carcinoma diagnosis. Further prospective studies should be conducted to highlight the theoretical strengths before its use in clinic.

[Infection of recombinant adenovirus Ad5F35-IL-12 in different kinds of human mononuclear macrophages].

Objective To investigate the efficiencies of transfection and expression of human recombinant adenovirus Ad5F35-IL-12 in the different kinds of human mononuclear macrophages. Methods The human recombinant adenovirus Ad5F35-IL-12 was used to infect human peripheral blood monocytes, pleural fluid macrophages as well as THP-1, U937 monocyte cell lines and their phorbol myristate acetate (PMA)-induced macrophages. 48 h later, green fluorescence was observed under the fluorescence microscope to detect the transfection efficiency. The expressions of IL-12 double-subunits (p35, p40) mRNA were tested by RT-PCR and the level of IL-12p70 protein in the cell culture supernatant was detected with ELISA. Results The human recombinant adenovirus Ad5F35-IL-12 successfully infected the human peripheral blood monocytes, pleural fluid macrophages, THP-1 monocytes, U937 monocytes, and THP-1 and U937 macrophages induced with PMA. All above infected mononuclear macrophages effectively secreted IL-12p70 protein, and they were listed from high to low of IL-12p70 protein level as pleural fluid macrophages, U937 and THP-1 macrophages induced with PMA, U937 monocytes, human peripheral blood monocytes and THP-1 monocytes. Conclusion The human recombinant adenovirus Ad5F35-IL-12 could infect different kinds of mononuclear macrophages, and IL-12 p70 protein could be successfully expressed in cell supernatants.

[Prokaryotic expression and purification of a tandem repeat of ovarian cancer antigen CA125 and preparation of its antiserum].

AIM: To establish a prokaryotic expression system of the tandem repeat of CA125 (CA125R), express and purify the recombinant CA125R protein, prepare its antiserum. METHODS: The full gene sequence of one tandem repeat of CA125 was synthesized and cloned into pET-32a(+) to construct a prokaryotic expression vector of the CA125R protein (pET-CA125R). The pET-CA125R was transformed into E.coli BL21 (DE3) and the soluble expression conditions were optimized; the pure recombinant CA125R protein was prepared by affinity Ni-NTA chromatography and identified by Western blotting. A rabbit was immunized with the pure recombinant CA125R protein to prepare its antiserum. RESULTS: The prokaryotic expression vector of CA125R was successfully constructed. The optimal soluble induction expression conditions were 0.5 mmol/L isopropyl ß-D-1-thiogalactopyranoside (IPTG) at 15DegreesCelsius for 6 h. Western blotting confirmed the pure CA125R recombinant protein of high purity. The prepared antiserum specifically recognized recombinant CA125R protein and natural CA125 glycoprotein. CONCLUSION: We successfully established the efficient prokaryotic expression system of the CA125R, and prepared the recombinant CA125R protein of high purity and its antiserum.