An Introduced RNA-Only Approach for Plasmid Curing via the CRISPR-Cpf1 System in Saccharomyces cerevisiae.

The CRISPR-Cas system has been widely used for genome editing due to its convenience, simplicity and flexibility. Using a plasmid-carrying Cas protein and crRNA or sgRNA expression cassettes is an efficient strategy in the CRISPR-Cas genome editing system. However, the plasmid remains in the cells after genome editing. Development of general plasmid-curing strategies is necessary. Based on our previous CRISPR-Cpf1 genome-editing system in Saccharomyces cerevisiae, the crRNA, designed for the replication origin of the CRISPR-Cpf1 plasmid, and the ssDNA, as a template for homologous recombination, were introduced for plasmid curing. The efficiency of the plasmid curing was 96 ± 4%. In addition, we further simplified the plasmid curing system by transforming only one crRNA into S. cerevisiae, and the curing efficiency was about 70%. In summary, we have developed a CRISPR-mediated plasmid-curing system. The RNA-only plasmid curing system is fast and easy. This plasmid curing strategy can be applied in broad hosts by designing crRNA specific for the replication origin of the plasmid. The plasmid curing system via CRISPR-Cas editing technology can be applied to produce traceless products without foreign genes and to perform iterative processes in multiple rounds of genome editing.

Assessment of the impact of climate change on endangered conifer tree species by considering climate and soil dual suitability and interspecific competition.

Climate change results in the habitat loss of many conifer tree species and jeopardizes species biodiversity and forest ecological functions. Delineating suitable habitats for tree species via climate niche model (CNM) is widely used to predict the impact of climate change and develop conservation and management strategies. However, the robustness of CNM is broadly debated as it usually does not consider soil and competition factors. Here we developed a new approach to combine soil variables with CNM and evaluate interspecific competition potential in the niche overlapping areas. We used an endangered conifer species - Chamaecyparis formosensis (red cypress) - as a case study to predict the impact of climate change. We developed a novel approach to integrate the climate niche model and soil niche model predictions and considered interspecific competition to predict the impacts of climate change on tree species. Our results show that the suitable habitat for red cypress would decrease significantly in the future with an additional threat from the competition of an oak tree species. Our approach and results may represent significant implications in making conservation strategies and evaluating the impacts of climate change, and providing the direction of the refinement of the ecological niche model.

Deletion of NTH1 and HSP12 increases the freeze-thaw resistance of baker's yeast in bread dough.

BACKGROUND: The intracellular molecule trehalose in Saccharomyces cerevisiae may have a major protective function under extreme environmental conditions. NTH1 is one gene which expresses trehalase to degrade trehalose. Small heat shock protein 12 (HSP12 expressed) plays a role in protecting membranes and enhancing freezing stress tolerance. RESULTS: An optimized S. cerevisiae CRISPR-Cpf1 genome-editing system was constructed. Multiplex genome editing using a single crRNA array was shown to be functional. NTH1 or/and HSP12 knockout in S. cerevisiae enhanced the freezing stress tolerance and improved the leavening ability after freezing and thawing. CONCLUSIONS: Deleting NTH1 in the combination with deleting HSP12 would strengthen the freezing tolerance and protect the cell viability from high rates of death in longer-term freezing. It provides valuable insights for breeding novel S. cerevisiae strains for the baking industry through a more precise, speedy, and economic genome-editing system.

A novel diagnostic tool for hyaline vascular Castleman disease versus lymphoma based on contrast-enhanced computed tomography in neck mass.

BACKGROUND: Castleman disease and lymphoma each have a distinct treatment plan; however, they share the same features on contrast-enhanced computed tomography. METHODS: To assess the quantitative outcomes of Castleman disease versus lymphoma using contrast-enhanced computed tomography based on Hounsfield units (HU). We retrospectively reviewed eight patients with unicentric Castleman disease and 30 patients with lymphoma based on pathological diagnosis at China Medical University Hospital between 2015 and 2020. Preoperative computed tomography with contrast scans was reviewed, and the HU of each tumor were measured. RESULTS: This study included eight patients with unicentric Castleman disease (four men and four women; mean age, 33 years) and 25 patients with lymphoma (11 men and 14 women; mean age, 53 years). There was no significant difference in heterogeneity between the two diseases (0.161 ± 0.052 vs 0.239 ± 0.063, p = 0.22); however, enhancement in Castleman disease was higher than that in lymphoma (126.40 ± 31.90 vs 74.19 ± 7.11, p < 0.001), providing a very good diagnostic tool (cutoff point at 88.5-91.3, sensitivity 0.86/specificity 0.88). Furthermore, we found a highly linear relationship in Castleman disease, which was not noted in lymphoma. CONCLUSION: The value of HU provides a good diagnostic tool for the differential diagnosis of Castleman disease versus lymphoma in the neck lymph nodes. Considering the linear relationship in Castleman disease, an increasingly accurate differential diagnosis can be made.

Food Safety Assessment of Commercial Genetically Modified Soybeans in Rats.

Although the safety of commercial genetically modified (GM) soybeans has been well evaluated and GM soybeans are legally sold under government management, some consumers still have concerns about their safety. The objective of this study was to evaluate the safety of commercial GM soybeans sold in markets as a food source. In the present study, two commercial GM (GM-1 and -2) soybeans and one non-GM soybean were randomly purchased and subjected to a whole food toxicity assessment. Rats (SD), male and female, were divided into six groups (10/sex/group). Two dosages of 1 g/kg/day and 5 g/kg/day of soybeans were selected for the low- and high-dose groups. Rats were administered the soybeans via daily oral fed for 90 days. The results indicate that the body weight, organ weight, biochemistry, hematology, and urology showed no biologically adverse effects. At necropsy, no significant differences between organ weights were noted between the non-GM- and GM soybeans-treated groups. Moreover, no gross or histopathological lesions were observed in the high-dosage (5 g/kg/day) fed groups of the non-GM and GM soybean fed rats. In conclusion, this food safety assessment revealed that commercial GM soybeans are substantially equivalent to non-GM soybeans in rats.

Increased Risk of Postpartum Depression in Women With Allergic Rhinitis During Pregnancy: A Population-Based Case-Control Study.

OBJECTIVES: Allergic rhinitis (AR) is associated with increased risk of major depression in the general population, however, no previous study has evaluated its role among pregnant women. We aimed to investigate the potential impact of AR during pregnancy on the development of postpartum depression (PPD). METHODS: This is a population-based case-control study. Data were retrieved from the National Health Insurance Research Database (NHIRD). Medical records of a total of 199 470 deliveries during 2000 and 2010 were identified. Among which, 1416 women with PPD within 12 months after delivery were classified as the case group, while 198 054 women without PPD after delivery formed the control group. Univariate and multivariate regression analyses were conducted to determine the associations between AR during pregnancies and other study variables with PPD. RESULTS: AR during pregnancy was found in 9.53% women who developed PPD and 5.44% in women without PPD. After adjusting for age at delivery, income level, various pregnancy and delivery-related conditions, asthma, atopic dermatitis and other medical comorbidities in the multivariate analysis, AR was significantly associated with increased odds of PPD (aOR: 1.498, 95% CI: 1.222-1.836). CONCLUSION: AR during pregnancy was independently and significantly associated with an approximately 50% increased risk of PPD among women giving birth. Closely monitoring of AR is warranted in the future in order to optimize mother and child outcomes after delivery.

Comparison of Findings between Clinical Examinations and Drug-Induced Sleep Endoscopy in Patients with Obstructive Sleep Apnea Syndrome.

The Velum, Oropharynx, Tongue base and Epiglottis (VOTE) classification on drug-induced sleep endoscopy (DISE) is used widely for obstructive sleep apnea (OSA) syndrome, though research into comparative physical examinations with VOTE on DISE is still limited. The aim of this study was to evaluate the relationship between the findings of physical examinations and DISE in patients with OSA. Fifty-five patients with OSA were enrolled in this retrospective study. All of the patients received clinical explorations including a Brodsky classification, a modified Mallampati score (MMS), a modified Friedman's staging system, and a Muller's test. Drug-induced sleep endoscopy was further evaluated in the operating room. There were significant relationships between Brodsky classification, modified Friedman's staging system, Muller's test and oropharynx collapse during DISE (p < 0.05). Brodsky classification, MMS, modified Friedman's staging system and retropalatal lateral-to-lateral (L-L) collapse of Muller's test were significantly correlated with VOTE count (p < 0.05). The concordance between VOTE under DISE and Brodsky classification or modified Friedman's staging system was moderate. In contrast, the concordance between VOTE under DISE and MMS or Muller's test was slight. The study revealed that Brodsky classification and Friedman staging had a significant relationship with DISE on the velum and oropharynx, but the level of tongue base is uncertain between DISE and MMS. Correlation of awake evaluation of tongue base is still not correlated to the DISE findings. Pre-treatment evaluation of DISE is still warranted.

[Status, problems and warranty strategy of quality uniformity for traditional Chinese medicine preparations].

The quality uniformity of traditional Chinese medicine (TCM) preparation is the base for guaranteeing the safety and effectiveness of clinical medication. At present, the quality of TCM preparation is uneven. At present, the same TCM preparation in different manufacturers, TCM preparations in the same manufacturer, and even different batches of a same TCM preparation in the same manufacturer have great differences in quality, which can not reach stability and uniformity. This paper would discuss the possible factors that influence the uniformity of quality in the whole process of pharmacy by means of consulting relevant literature on quality control of Chinese herbal preparations and analyzing the present situation and problems of the quality of TCM preparation. In addition, some strategies such as standardization of cultivation of TCM, processing standardization, standardization of pharmaceutical equipment, mixed batch feeding, and Quality by Design would be also put forward to provide references for the quality uniformity of TCM preparation.

Isolation of 16 polymorphic microsatellite markers from an endangered and endemic species, Podocarpus nakaii (Podocarpaceae).

PREMISE OF THE STUDY: Sixteen polymorphic microsatellite markers were isolated and characterized from the endangered evergreen tree Podocarpus nakaii to evaluate the population structure for conservation efforts. METHODS AND RESULTS: Based on a modified amplified fragment length polymorphism and magnetic bead enrichment method, 16 polymorphic primer sets were developed for this endangered insular species. Allele numbers ranged from five to seven, with an observed heterozygosity ranging from 0.29 to 0.88. Most primers were able to amplify DNA from the endemic P. fasciculus and the widely distributed P. macrophyllus var. macrophyllus, P. macrophyllus var. maki, and P. costalis. CONCLUSIONS: The results reported here indicate the usefulness of codominant markers for future studies of the population genetics of P. nakaii. In addition, the markers are useful for further phytogeographic and speciation studies in P. fasciculus, P. macrophyllus var. macrophyllus, and P. macrophyllus var. maki, which are closely related species.

SOCS6, down-regulated in gastric cancer, inhibits cell proliferation and colony formation.

Members of the suppressor of cytokine-induced signaling (SOCS) family are negative regulators of cytokine signaling pathways. By mRNA differential display, we showed that SOCS6 was frequently down-regulated in gastric cancer (GC). Our data showed that allelic loss and promoter hypermethylation may account for the major mechanisms leading to SOCS6 inactivation. Ectopic expression of SOCS6 suppressed cell growth and colony formation, in part through eliciting intrinsic apoptotic pathway, accompanied with decreased mitochondrial membrane potential. Taken together, this study provides molecular and functional data supporting the importance of loss-of-function of SOCS6 as a frequent event in gastric tumorigenesis.

Telemetric electrochemical sensor.

A telemetric system was designed and constructed to sense pH and ethanol variation in aqueous solutions. The measured signals were transferred by software digitally and transmitted wirelessly by the telemeter, personal digital assistant (PDA), through the General Packet Radio Service (GPRS) protocol. The pH sensing electrode was designed to measure a chemical potential induced by a proton concentration gradient on the electrode's surface which exhibits internal Donnon diffusion behavior, and a linear relationship between the electrical potential and pH was found. The result shows that the wireless sensing system allowed not only long-term usage and long-distance transmission but also with high accuracy (e.g. S.D. less than +/-2%). The telemetric system can also be modified to measure ethanol concentration in aqueous solution amperometrically. It was found that the sensitivity of that ex situ measurements matched those of in field measurements with negligible deviation, less than 4%.