ID3 regulates progesterone synthesis in bovine cumulus cells through modulation of mitochondrial function.

DNA binding inhibitory factor 3 (ID3) has been shown to have a key role in maintaining proliferation and differentiation. It has been suggested that ID3 may also affect mammalian ovarian function. However, the specific roles and mechanisms are unclear. In this study, the expression level of ID3 in cumulus cells (CCs) was inhibited by siRNA, and the downstream regulatory network of ID3 was uncovered by high-throughput sequencing. The effects of ID3 inhibition on mitochondrial function, progesterone synthesis, and oocyte maturation were further explored. The GO and KEGG analysis results showed that after ID3 inhibition, differentially expressed genes, including StAR, CYP11A1, and HSD3B1, were involved in cholesterol-related processes and progesterone-mediated oocyte maturation. Apoptosis in CC was increased, while the phosphorylation level of ERK1/2 was inhibited. During this process, mitochondrial dynamics and function were disrupted. In addition, the first polar body extrusion rate, ATP production and antioxidation capacity were reduced, which suggested that ID3 inhibition led to poor oocyte maturation and quality. The results will provide a new basis for understanding the biological roles of ID3 as well as cumulus cells.

TBX2 affects proliferation, apoptosis and cholesterol generation by regulating mitochondrial function and autophagy in bovine cumulus cell.

BACKGROUND: T-box transcription factor 2 (TBX2) is a member of T-box gene family whose members are highly conserved in evolution and encoding genes and are involved in the regulation of developmental processes. The encoding genes play an important role in growth and development. Although TBX2 has been widely studied in cancer cell growth and development, its biological functions in bovine cumulus cells remain unclear. OBJECTIVES: This study aimed to investigate the regulatory effects of TBX2 in bovine cumulus cells. METHODS: TBX2 gene was knockdown with siRNA to clarify the function in cellular physiological processes. Cell proliferation and cycle changes were determined by xCELLigence cell function analyzer and flow cytometry. Mitochondrial membrane potential and autophagy were detected by fluorescent dye staining and immunofluorescence techniques. Western blot and quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) were used to detect the expression changes of proliferation and autophagy-related proteins. Aadenosine triphosphate (ATP) production, glucose metabolism, and cholesterol synthesis of cumulus cells were measured by optical density and chemiluminescence analysis. RESULTS: After inhibition of TBX2, the cell cycle was disrupted. The levels of apoptosis, ratio of light chain 3 beta II/I, and reactive oxygen species were increased. The proliferation, expansion ability, ATP production, and the amount of cholesterol secreted by cumulus cells were significantly decreased. CONCLUSIONS: TBX2 plays important roles in regulating the cells' proliferation, expansion, apoptosis, and autophagy; maintaining the mitochondrial function and cholesterol generation of bovine cumulus cells.

Prevalence and spatial distribution patterns of human echinococcosis at the township level in Sichuan Province, China.

BACKGROUND: Echinococcosis is a global zoonotic parasitic disease caused by Echinococcus larvae. This disease is highly endemic in Sichuan Province, China. This study investigates the prevalence and spatial distribution characteristics of human echinococcosis at the township level in Sichuan Province, geared towards providing a future reference for the development of precise prevention and control strategies. METHODS: Human prevalence of echinococcosis was evaluated using the B-ultrasonography diagnostic method in Sichuan Province between 2016 and 2019. All data were collected, collated, and analyzed. A spatial distribution map was drawn to intuitively analyze the spatial distribution features. Eventually, the spatial autocorrelation was specified and local indicators of spatial association (LISA) clustering map was drawn to investigate the spatial aggregation of echinococcosis at the township level in Sichuan Province. RESULTS: The prevalence of echinococcosis in humans of Sichuan Province was 0.462%, among which the occurrence of cystic echinococcosis (CE) was 0.221%, while that of alveolar echinococcosis (AE) was 0.244%. Based on the results of the spatial distribution map, a predominance of echinococcosis in humans decreased gradually from west to east and from north to south. The Global Moran's I index was 0.77 (Z = 32.07, P < 0.05), indicating that the prevalence of echinococcosis in humans was spatially clustered, exhibiting a significant spatial positive correlation. Further, the findings of local spatial autocorrelation analysis revealed that the "high-high" concentration areas were primarily located in some townships in the northwest of Sichuan Province. However, the "low-low" concentration areas were predominantly located in some townships in the southeast of Sichuan Province. CONCLUSIONS: Our findings demonstrated that the prevalence of echinococcosis in humans of Sichuan Province follows a downward trend, suggesting that the current prevention and control work has achieved substantial outcomes. Nevertheless, the prevalence in humans at the township level is widely distributed and differs significantly, with a clear clustering in space. Therefore, precise prevention and control strategies should be formulated for clusters, specifically strengthening the "high-high" clusters at the township level.

Prophylactic Central Neck Dissection for cN1b Papillary Thyroid Carcinoma: A Systematic Review and Meta-Analysis.

BACKGROUND: The value of prophylactic central neck dissection (PCND) for papillary thyroid carcinoma (PTC) with clinically evident lateral cervical lymph node metastases (cN1b) remains unclear. Therefore, a systematic review and meta-analysis was conducted to assess the efficacy and safety of PCND. METHODS: A comprehensive systematic search was conducted on PubMed, Web of Science, Cochrane library and Embase databases up to September 2021 to identify eligible studies. Controlled clinical trials assessing therapeutic effects and safety of PCND for cN1b PTC patients were included. The risk of bias for each cohort study was assessed using the Newcastle-Ottawa Scale (NOS). The primary outcomes were indexes related to the locoregional recurrence (LRR) and surgical complications. Review Manager software V5.4.0 was used for statistical analysis. A fixed effects model was adopted for the data without heterogeneity, otherwise a random effects model was used. RESULTS: We included 4 retrospective cohort studies, which comprised 483 PTC patients. There was no statistically significant difference in the central neck recurrence (CNR) (10.2% vs. 3.8%, relative risk (RR) = 1.82; 95%CI 0.90-3.67; P = 0.09), lateral neck recurrence (LNR) (5.1% vs. 7.7%, RR = 0.47; 95% CI 0.13-1.74; P = 0.26), and overall recurrence (OR) (18.9% vs. 16.9%, RR = 0.77; 95%CI 0.34-1.76; P = 0.54), between LND + PCND group and LND group. Simultaneously, PCND increased the risk of permanent hypoparathyroidism (11.4% vs. 4.5%, RR = 2.70, 95%CI 1.05-6.94; P = 0.04) and overall complications (17.0% vs. 5.3%, RR = 3.28; 95%CI 1.37-7.86; P = 0.008). CONCLUSIONS: This meta-analysis showed that PCND did not have any advantage in preventing LRR for cN1b PTC. Meanwhile, PCND may result in the increased rate of surgical complications. However, the current evidence is limited and more clinical trials are still needed to further clarify the true role of PCND. SYSTEMATIC REVIEW REGISTRATION: https://www.crd.york.ac.uk/PROSPERO/, CRD42021281825.

Carnosic acid improves porcine early embryonic development by inhibiting the accumulation of reactive oxygen species.

Carnosic acid (CA), a natural catechol rosin diterpene, is used as an additive in animal feeds and human foods. However, the effects of CA on mammalian reproductive processes, especially early embryonic development, are unclear. In this study, we added CA to parthenogenetically activated porcine embryos in an in vitro culture medium to explore the influence of CA on apoptosis, proliferation, blastocyst formation, reactive oxygen species (ROS) levels, glutathione (GSH) levels, mitochondrial membrane potential, and embryonic development-related gene expression. The results showed that supplementation with 10 µM CA during in vitro culture significantly improved the cleavage rates, blastocyst formation rates, hatching rates, and total numbers of cells of parthenogenetically activated porcine embryos compared with no supplementation. More importantly, supplementation with CA also improved GSH levels and mitochondrial membrane potential, reduced natural ROS levels in blastomeres, upregulated Nanog, Sox2, Gata4, Cox2, Itga5, and Rictor expression, and downregulated Birc5 and Caspase3 expression. These results suggest that CA can improve early porcine embryonic development by regulating oxidative stress. This study elucidates the effects of CA on early embryonic development and their potential mechanisms, and provides new applications for improving the quality of in vitro-developed embryos.

Disease burden of echinococcosis in Tibetan communities-A significant public health issue in an underdeveloped region of western China.

Echinococcosis is considered by the World Health Organization (WHO) to be a neglected zoonotic disease in the world. Some Tibetan communities were found to be highly endemic for echinococcosis just 20 years ago. Until recently, we were able to understand the overall disease burden of echinococcosis in Tibetan communities after prevalence data being available from nationwide investigations from 2012 to 2016. Data were abstracted from 9 publications regarding to echinococcosis prevalence between 2016-2018; from 10 data bases on echinococcosis prevalence for 151 Tibetan counties; and statistics of population, Gross Domestic Product (GDP) and health staff from 44 local statistic bureaus and government websites at provincial, prefecture and county level, and 2 books of provincial yearly statistics. These data were used to estimate the Disability Adjusted Life Years (DALYs) due to cystic echinococcosis (CE) and alveolar echinococcosis (AE). The distribution of DALYs was presented geographically and economically. The echinococcosis DALYs in the Tibetan communities were estimated to be 126,159 (95%UI 122,415-137,675) annually using the method recommended by WHO. AE DALYs were estimated to be 105,829 (95%UI 101,969-117,090), which were more than CE DALYs of 20,330 (95%UI 19,690-21,581). Echinococcosis affects people more in underdeveloped areas. There was a tendency that a higher echinococcosis DALYs were usually correlated a higher altitude. Health services are also poorly provided in terms of number of health staff of 5.05 per 1000 population in comparison with the national average of 5.8 per 1000 population. The data suggest that the echinococcosis burden in the center region of Qinghai-Tibet plateau is higher than that of other regions, and consequently more control and health services should be provided to the region.

Self-Assembled Peptide Nanostructures for Photoelectrochemical Bioanalysis Application: A Proof-of-Concept Study.

Currently, one of important research directions of photoelectrochemical (PEC) bioanalysis is to exploit innovative photoactive species and their elegant implementations for selective detection and signal transduction. Different from existing candidates for photoelectrode development, this study, exemplified by the cationic dipeptide nanoparticles (CDNPs), reports the first demonstration of self-assembled peptide nanostructures (SAPNs) for the PEC bioanalysis. Specifically, the CDNPs were prepared as representative materials and then immobilized onto the indium tin oxide (ITO) electrode for the PEC differentiation of several commonly involved biomolecules such as ascorbic acid (AA) and l-cysteine. Significantly, the experimental results disclosed that the CDNPs possessed unique photocathodic responses and good analytical performance toward AA detection in terms of rapid response, high stability, and excellent selectivity. This work demonstrates the great potential of the large SAPN family for the future PEC bioanalysis development and has not been reported to our knowledge.

A multiplex PCR for differential detection of Echinococcus granulosus sensu stricto, Echinococcus multilocularis and Echinococcus canadensis in China.

BACKGROUND: Echinococcosis caused by Echinococcus is one of the most major infectious diseases in north-west highland of China. E. granulosus sensu strict, E. multilocularis, and E. canadensis are known to be the only three species related to human health transmitting in the areas. To achieve targeted treatment and control of echinococcosis, the accurate identification and discrimination of the species are important. However, currently the available diagnostic approaches do not present ideal results either in accuracy or efficiency. METHODS: In the study, a set of primers were designed to aim at the three human-pathogenic Echinococcus species in China. The one-step multiplex PCR assay was developed and evaluated for the specificity and sensitivity. A total of 73 parasitic lesions and 41 fecal materials obtained from human and various animals collected in the clinic and the field were tested to assess the applicability of this method. RESULTS: The multiplex PCR effectively detected the individual DNA from the targeted species and their random mixtures generating with distinguishable expected size of products. The detection limit of the assay for each of the three species was 5 pg/µl when they were tested separately. When DNA mixtures of the targeted species containing the same concentration were used as templates, the lowest amount of DNA which can be detected was 50 pg/µl, 10 pg/µl and 5 pg/µl for E. granulosus s. s., E. multilocularis, and E. canadensis respectively. No cross-reactivity was observed when DNA from eight genetically close species was used as control templates. The multiplex PCR identifications of all samples were in line with the original sequencing results except for those infected with E. shiquicus, which showed negative signals in the developed assay. Of all the tested stool materials, 16 were previously found positive for Echinococcus by visual and microscopic examination. Among these 16 samples, 13 were confirmed by the multiplex PCR, and the other three tested negative. Additionally, the multiplex PCR identified another 14 positive feces from the remained 25 stool samples which absence of worms. CONCLUSIONS: The developed multiplex PCR shows advantages in fast diagnosis and large-scale epidemiological investigation, which proven to be a promising tool utilized in clinic and surveillance system.

Geographic distribution of echinococcosis in Tibetan region of Sichuan Province, China.

BACKGROUND: Echinococcosis is a parasitic zoonosis caused by Echinococcus larvae parasitism causing high mortality. The Tibetan Region of Sichuan Province is a high prevalence area for echinococcosis in China. Understanding the geographic distribution pattern is necessary for precise control and prevention. In this study, a spatial analysis was conducted to explore the town-level epidemiology of echinococcosis in the Sichuan Tibetan Region and to provide guidance for formulating regional prevention and control strategies. METHODS: The study was based on reported echinococcosis cases by the end of 2017, and each case was geo-coded at the town level. Spatial empirical Bayes smoothing and global spatial autocorrelation were used to detect the spatial distribution pattern. Spatial scan statistics were applied to examine local clusters. RESULTS: The spatial distribution of echinococcosis in the Sichuan Tibetan Region was mapped at the town level in terms of the crude prevalence rate, excess hazard and spatial smoothed prevalence rate. The spatial distribution of echinococcosis was non-random and clustered with the significant global spatial autocorrelation (I = 0.7301, P = 0.001). Additionally, five significant spatial clusters were detected through the spatial scan statistic. CONCLUSIONS: There was evidence for the existence of significant echinococcosis clusters in the Tibetan Region of Sichuan Province, China. The results of this study may assist local health departments with developing better prevention strategies and prompt more efficient public health interventions.

Identification of Polar Constituents in the Decoction of Juglans mandshurica and in the Medicated Egg Prepared with the Decoction by HPLC-Q-TOF MS².

As a folk medicinal plant, Juglans mandshurica has been used for the treatment of cancer in China and Korea. Traditionally, J. mandshurica is decocted together with chicken eggs. Both the decoction and medicated eggs possess anti-tumor properties. Clarifying the constituents of the decoction and absorbed by the medicated eggs is essential for the investigation of the active principles of J. mandshurica. Herein, the medicated eggs were prepared by decocting raw chicken eggs, having unbroken shells, with the decoction of J. mandshurica. A systematic investigation of the chemical profile of the J. mandshurica decoction and the medicated egg extraction was conducted by HPLC-Q-TOF-MS². In total, 93 peaks, including 45 tannins, 14 naphthalene derivatives, 17 organic acids, 3 diarylheptanoids, 4 lignans, 3 anthraquinones, 1 flavonoid glycoside, 3 amino acids, and 3 nitrogenous compounds, were tentatively identified in the decoction. In the medicated egg extraction, 44 peaks including 11 organic acids, 3 amino acids, 3 nitrogenous compounds, 8 naphthalene derivatives, 3 diarylheptanoids, 15 tannins, and 1 lignan were tentatively identified. The chemical profile presented provided a detailed overview of the polar chemical constituents in J. mandshurica and useful information for the research of bioactive compounds of this plant.

Seasonal pattern of Echinococcus re-infection in owned dogs in Tibetan communities of Sichuan, China and its implications for control.

BACKGROUND: Human cystic echinococcosis (CE) and alveolar echinococcosis (AE) are highly endemic in Tibetan communities of Sichuan Province. Previous research in the region indicated that domestic dog was the major source of human infection, and observations indicated that domestic dog could have more access to intermediate hosts of Echinococcus spp.: both domestic livestock (CE) viscera and small mammals (AE), in early winter and again in spring. We hypothesized that there would therefore be a significant increase in the risk of canine infection with Echinococcus spp. in these two seasons and conducted a reinfection study to investigate this further. METHODS: Faecal samples were collected from owned dogs in seven townships in Ganze Tibetan Autonomous Prefecture (Sichuan Province, China), and Echinococcus spp. infection status was determined using copro-antigen ELISA. Dogs were sampled in April (spring), July (early summer), September/October (autumn/early winter) and December (winter) in 2009; and in April (spring) 2010. Dogs were treated with praziquantel following each of the five sample collections to eliminate any tapeworms. Information on dog sex, age and body weight was also collected. The t-test, Fisher's exact test, Poisson regression and logistic regression were used to compare means and prevalences, and to identify factors associated with infection status. RESULTS: The proportion of female dogs was significantly lower than that of male dogs; female dogs had significantly higher (22.78 %) baseline copro-ELISA prevalence than males (11.88 %). Dog body weight, sex, age, county and previous infection status at any sampling point had no influence on the re-infection prevalence in general. Poisson regression did not found a significant influence on the re-infection prevalence due to different deworming/sampling time spans. Dogs exhibited significantly higher re-infection prevalences in spring and early summer of 2009 and in early winter between September/October and December of 2009, suggesting a higher infection pressure in these seasons comparing with other seasons. CONCLUSION: Following praziquantel treatment, dog body weight, sex, age, county, deworming time span and previous infection status at any sampling point had no influence on the re-infection prevalence in the region in general. The differences between re-infection prevalences were probably due to the seasonality in Echinoccocus spp. infection pressure in the region. Early winter, spring and early summer should be important seasons for optimal dog deworming intervention in these Tibetan communities.

[Comparison of biological behavior between triple-negative breast cancer and non-triple- negative breast cancer].

OBJECTIVE: To summarize the epidemiological and biological features of triple-negative breast cancer (TNBC) and non-triple-negative breast cancer (non-TNBC) to provide reference for devising individualized therapy and making prognostic evaluation. METHODS: The 5-year follow-up data were collected from 231 patients with pathologically established diagnosis of breast cancer treated in the Affiliated Hospital of Ningxia Medical University and Yinchuan People's Hospital between Jan. 2002 and Dec. 2004. The epidemiological and clinicopathological characteristics as well as the recurrence, metastasis and 5-year survival were compared between TNBC group and non-TNBC group. RESULTS: TNBC accounted 17.3% of the total breast cancer cases enrolled in this study. The tumor size and rates of recurrence and metastasis (especially visceral metastasis) were significantly greater in TNBC group than in non-TNBC group (P<0.05). The TNBC patients showed significantly lower 3- and 5-year survival rates than the non-TNBC patients (P<0.05), and TNBC patients with positive lymph nodes in clinical stage II had also a lower 5-year survival (P<0.05). Cox regression model analysis identified the patients' age, primary tumor size, clinical stages and triple-negativity as the independent risk factors for breast cancer. CONCLUSION: Compared to non-TNBC patients, patients with TNBC have higher rates of local recurrence and invasion, visceral metastasis and poorer prognosis, and a lower rate of 5-year survival. The triple negativity represents an independent factor for prognosis evaluation of breast cancer.