Revealing the dynamic whole transcriptome landscape of Clonorchis sinensis: Insights into the regulatory roles of noncoding RNAs and microtubule-related genes in development.

Clonorchis sinensis is a significant zoonotic food-borne parasite that causes a range of hepatobiliary diseases, which in severe cases can even lead to cholangiocarcinoma. To explore new diagnostic and treatment strategies, the dynamic RNA regulatory processes across different developmental stages of C. sinensis were analyzed by using whole-transcriptome sequencing. The chromosomal-level genome of C. sinensis was used for sequence alignment and annotation. In this study, we identified a total of 59,103 RNAs in the whole genome, including 2,384 miRNAs, 25,459 mRNAs, 27,564 lncRNAs and 3,696 circRNAs. Differential expression analysis identified 6,556 differentially expressed mRNAs, 2,231 lncRNAs, 877 miRNAs and 20 circRNAs at different developmental stages. Functional enrichment analysis highlighted the critical role of microtubule-related biological processes in the growth and development of C. sinensis. And coexpression analysis revealed 97 lncRNAs and 85 circRNAs that were coexpressed with 42 differentially expressed mRNAs that associated with microtubules at different developmental stages of C. sinensis. The expression of the microtubule-related genes dynein light chain 2 (DLC2) and dynein light chain 4 (DLC4) increased with C. sinensis development, and DLC2/4 could be inhibited by albendazole. Finally, by constructing competing endogenous RNA (ceRNA) networks, the lncRNA-miRNA-mRNA and circRNA-miRNA-mRNA regulatory relationships were constructed, and the ceRNA networks of MSTRG.14258.5-novel_miR_2287-newGene_28215 and MSTRG.14258.5-novel_miR_2216-CSKR_109340 were verified. This study suggests, through whole transcriptome sequencing, that the context of microtubule regulation may play an essential role in the development and growth of C. sinensis.

Agarose Hydrogel-Boosted One-Tube RPA-CRISPR/Cas12a Assay for Robust Point-of-Care Detection of Zoonotic Nematode Anisakis.

Rapid and accurate detection of the zoonotic nematode Anisakis is poised to control its epidemic. The clustered regularly interspaced short palindromic repeats (CRISPR)/Cas-associated assay shows great potential in the detection of pathogenic microorganisms. The one-tube method integrated the CRISPR system with the recombinase polymerase amplification (RPA) system to avoid the risk of aerosol pollution; however, it suffers from low sensitivity due to the incompatibility of the two systems and additional manual operations. Therefore, in the present study, the agarose hydrogel boosted one-tube RPA-CRISPR/Cas12a assay was constructed by adding the CRISPR system to the agarose hydrogel, which avoided the initially low amplification efficiency of RPA caused by the cleavage of Cas12a and achieved reaction continuity. The sensitivity was 10-fold higher than that of the one-tube RPA-CRISPR/Cas12a system. This method was used for Anisakis detection within 80 min from the sample to result, achieving point-of-care testing (POCT) through a smartphone and a portable device. This study provided a novel toolbox for POCT with significant application value in preventing Anisakis infection.

Relationship between urodynamic patterns and lower urinary tract symptoms in Chinese women with a non-neurogenic bladder.

OBJECTIVE: To investigate the urodynamic study (UDS) patterns, obstruction status, continence status, and their correlations among neurologically intact women with lower urinary tract symptoms (LUTS) through an epidemiological and logistic regression analyses. METHODS: We retrospectively analyzed the UDS data of 3265 neurologically intact women with LUTS (2002-2014). Five UDS patterns were identified: normo-active detrusor/sphincter (NA, or DSI, detrusor/sphincter intact), idiopathic detrusor overactivity (IDO), idiopathic sphincter overactivity (ISO), IDO + ISO, and detrusor underactivity (DUA). Analyses of UDS pattern distribution and stratification were performed (based on a modification of the European Urological Association-Madersbacher classification system), and their correlations with bladder outlet obstruction (BOO) and stress urinary incontinence (SUI) status were evaluated via logistic regression analysis. RESULTS: NA, IDO, IDO + ISO, ISO, and DUA were noted in 927 (28.4%), 678 (20.8%), 320 (9.8%), 689 (21.1%), and 651 (19.9%) cases, respectively. Moreover, storage, storage + voiding, and voiding symptoms were noted in 62.4%, 21.1%, and 16.5% cases, respectively, whereas BOO and SUI were observed in 12.1% and 29.0% cases, respectively. The risk factors for BOO included NA, IDO, ISO, and IDO + ISO, whereas the protective factors against BOO included storage symptoms, SUI, storage + voiding symptoms, and complaint duration within 1-12 months. NA was the only risk factor for SUI, whereas BOO, storage + voiding symptoms, IDO, and storage symptoms were protective factors for SUI. CONCLUSION: Five UDS patterns were identified among neurologically intact women with LUTS. Functional abnormalities of the detrusor and/or sphincter were the main causes of LUTS, and were correlated with the BOO or SUI status. Thus, the UDS pattern can provide additional information regarding the risk factors for BOO or SUI status, as compared to symptomatic typing.

Urodynamic pattern distribution among aged male patients with lower urinary tract symptoms suggestive of bladder outlet obstruction.

OBJECTIVE: To develop a urodynamic study (UDS) pattern system for aged male patients who complained of non-neurogenic lower urinary tract symptoms (LUTS) to create a reference guideline for their diagnosis and treatment by a retrospective analysis. METHODS: A retrospective analysis of UDS data was carried out in 1984 male patients neurologically intact with symptoms suggestive of bladder outlet obstruction (BOO) aged older than 45 years (2002-2013). On the basis of their UDS characteristic findings, the patients were classified into 1 of 7 subgroups: equivocal or mild BOO with sphincter synergia with or without idiopathic detrusor overactivity (pattern A); equivocal or mild BOO with idiopathic sphincter overactivity (B); classic BOO with sphincter synergia (C) or overactivity (D); BOO with only detrusor low compliance (E); BOO with both detrusor underactivity and low compliance (F); and equivocal BOO with detrusor underactivity (G). The follow-up data were reviewed and analyzed thereafter. RESULTS: The feasibility and rationality of this system were confirmed. The distribution of 7 patterns (pattern, case number, %) was A 158, 8%; B 59, 3%; C 1059, 53.3%; D 277, 14%; E 120, 6%; F 93, 4.7%; and G 218, 11%. A-G numbers in pattern C, D, and E were 103.1-141.4, higher than other patterns (P <.001), and functional pressure lengths of pattern C and D were 7.0-7.2 cm, longer than other patterns (P <.001). CONCLUSION: A practical UDS pattern system for aged male patients with lower urinary tract symptoms suggestive of BOO was constructed, which can be used to optimize the diagnosis and treatment of these patients.

Low-frequency electrotherapy for female patients with detrusor underactivity due to neuromuscular deficiency.

INTRODUCTION AND HYPOTHESIS: The aim of the study was to assess the efficacy of low-frequency electrotherapy (LFE) for female patients with early-stage detrusor underactivity (DUA) due to neuromuscular deficiency. METHODS: A total of 102 female patients were divided randomly into four groups: LFE-NC (normal compliance), LFE-LC (low compliance), CON (control)-NC and CON-LC. Patients in the LFE-NC and LFE-LC groups received LFE, and those in the CON-NC and CON-LC groups received conservative treatment. Urodynamic evaluation was performed before and after treatment. RESULTS: After treatment, 82 % of the LFE-NC regained detrusor contractility, whereas only 2 (8 %) of the CON-NC had normal detrusor contraction. None of LFE-LC or CON-LC regained detrusor contractility (p < 0.01). The per cent of LFE-NC who relied on catheterization for bladder emptying decreased by 43 % (p < 0.01). Those in the LFE-LC, CON-NC and CON-LC groups decreased by only 4, 12 or 0 % (p > 0.05). CONCLUSIONS: LFE was more effective for DUA patients with normal compliance; these patients benefited from LFE, but DUA patients with low compliance did not.