MLL-AF9 regulates transcriptional initiation in mixed lineage leukemic cells.

MLL-fusion proteins (MLL-FPs) are believed to maintain gene activation and induce mixed lineage leukemia (MLL) through aberrantly stimulating transcriptional elongation, but the underlying mechanisms are incompletely understood. Here we show that both MLL1 and AF9, one of the major fusion partners of MLL1, mainly occupy promoters and distal intergenic regions, exhibiting chromatin occupancy patterns resembling that of RNA polymerase II (Pol II) in HEL, a human cell line without MLL1 arrangement (MLLr). MLL1 and AF9 only co-regulate over a dozen genes despite of their co-occupancy on thousands of genes. They do not interact with each other, and their chromatin occupancy is also independent of each other. Moreover, AF9 deficiency in HEL cells decreases global TBP occupancy while decreases CDK9 occupancy on a small number of genes, suggesting an accessory role of AF9 in CDK9 recruitment and a possible major role in transcriptional initiation via initiation factor recruitment. Importantly, MLL1 and MLL-AF9 occupy promoters and distal intergenic regions, exhibiting identical chromatin occupancy patterns in MLL cells, and MLL-AF9 deficiency decreased occupancy of TBP and TFIIE on major target genes of MLL-AF9 in iMA9, a murine acute myeloid leukemia (AML) cell line inducibly expressing MLL-AF9, suggesting that it can also regulate initiation. These results suggest that there is no difference between MLL1 and MLL-AF9 with respect to location and size of occupancy sites, contrary to what people have believed, and that MLL-AF9 may also regulate transcriptional initiation in addition to widely-believed elongation.

AFF4 globally affects the release of paused RNA polymerase II in HEL cells.

P-TEFb, a heterodimer of the kinase CDK9 and Cyclin T1, is a critical regulator of promoter-proximal pause release of Pol II in metazoans. It is capable of forming three larger complexes, including the super elongation complex (SEC), the BRD4/P-TEFb complex and the 7SK snRNP. In the SEC or the BRD4/P-TEFb complex, P-TEFb is enzymatically active, while in the 7SK snRNP, its activity is inhibited. The SEC consists of AFF1 or 4, ENL or AF9, ELL1, 2 or 3 and EAF1 or 2 in addition to P-TEFb, the only subunit with catalytic activity, and the noncatalytic subunits have been found to be able to regulate pause release through P-TEFb. We and others recently found that AFF1, ENL and AF9 are capable of regulating transcriptional initiation, but it is unknown yet whether AFF4 is also capable of doing so. With respect to the gene regulation selectivity of the SEC and the BRD4/P-TEFb complex, one recent study showed that in human DLD-1 cells, the SEC only regulates pause release of heat shock (HS) genes, whereas the BRD4/P-TEFb complex regulates pause release of the rest of the genes. However, it is unclear whether those mechanisms are general. In this study for the purpose of further understanding the role of AFF4 in transcriptional regulation, we found that AFF4 knockdown by RNA interference in human HEL cells decreased not only cellular level but also global chromatin occupancy of CTD serine 2 phosphorylated Pol II. Direct target genes of AFF4 were identified by RNA-seq and CUT&Tag. Notably, we found by ChIP-seq and PRO-seq that AFF4 loss also increased promoter-proximal pause of Pol II on several hundred HS and thousands of non-HS genes. Mechanistically, AFF4 promotes pause release likely by facilitating the binding of P-TEFb to Pol II. These results suggest that extent of the impact of AFF4 on pause release is likely to be context-dependent or cell-type dependent.

Tox4 regulates transcriptional elongation and reinitiation during murine T cell development.

HMG protein Tox4 is a regulator of PP1 phosphatases with unknown function in development. Here we show that Tox4 conditional knockout in mice reduces thymic cellularity, partially blocks T cell development, and decreases ratio of CD8 to CD4 through decreasing proliferation and increasing apoptosis of CD8 cells. In addition, single-cell RNA-seq discovered that Tox4 loss also impairs proliferation of the fast-proliferating double positive (DP) blast population within DP cells in part due to downregulation of genes critical for proliferation, notably Cdk1. Moreover, genes with high and low expression level are more dependent on Tox4 than genes with medium expression level. Mechanistically, Tox4 may facilitate transcriptional reinitiation and restrict elongation in a dephosphorylation-dependent manner, a mechanism that is conserved between mouse and human. These results provide insights into the role of TOX4 in development and establish it as an evolutionarily conserved regulator of transcriptional elongation and reinitiation.

GeneChip expression profiling identified OLFML2A as a potential therapeutic target in TNBC cells.

Background: An elevated level of olfactomedin-like-2A (OLFML2A) is unfavorable for female breast cancer patients. Patients with a high mRNA level of OLFML2A receive a poor prognosis. Therefore, we speculate that inhibiting the expression of this gene may be beneficial to breast cancer patients. We previously found that silencing the OLFML2A gene by using mRNA interference significantly inhibited proliferation and migration in triple-negative breast cancer (TNBC) cells. Methods: Cell activity and proliferation were determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and Celigo analyses. Cell migration and invasion were determined by wound-healing and transwell invasion assays. The mechanism of the inhibition of a small hairpin RNA that targets OLFML2A (shOLFML2A) was determined by using a GeneChip array, real-time quantitative PCR (RT-qPCR), and western blot analysis. Results: Gene silencing by shOLFML2A induces apoptosis by promoting S phase arrest in TNBC cells. In addition, shOLFML2A decreased the progression of epithelial-mesenchymal transition (EMT). Additionally, microarray analysis showed that shOLFML2A significantly upregulated 428 genes and downregulated 712 genes. These significantly changed genes regulated DNA synthesis, chromosome alignment, microtubules and the cytoskeleton, cell movement, the cell cycle, cell necrosis, and apoptosis because they promoted G2/M DNA damage checkpoint regulation and p53 signaling, and because they inhibited integrin, hepatocyte growth factor (HGF), nerve growth Factor (NGF), and other tumor-promoting signaling pathways. Conclusions: shOLFML2A reduces cell proliferation, migration, and invasion and promotes cell apoptosis. Therefore, the results of the present study suggest that OLFML2A is a potential therapeutic target for TNBC.

Amorphophalli Rhizoma inhibits breast cancer growth, proliferation, migration, and invasion via the PI3K/AKT pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Amorphophalli Rhizoma (APR) is widely used as an adjuvant treatment for advanced and metastatic triple-negative breast cancer (TNBC), but its effects, potential active ingredients, and mechanism of action on estrogen receptor-positive (ER+) and human epidermal growth factor receptor-positive (HER2+) breast cancer cells were not reported. AIM OF THE STUDY: The present study investigated the effects and mechanism of APR on ER+ and HER2+ breast cancer cells. MATERIALS AND METHODS: Rotary evaporation was used to prepare different extracts of APR. Cell activity was assessed using the cell counting kit-8 (CCK-8) method. Wound healing assays were used to assess cell migration, and a cell invasion assay was performed using a Transwell chamber with Matrigel matrix. A xenograft model was used to analyze the inhibitory effects of APR on tumor growth. Bioinformatics analyses were used to explore the potential mechanism of APR in breast cancer. RT-qPCR and Western blotting were performed to reveal the molecular mechanism. RESULTS: The ethyl acetate extract of APR showed the strongest tumor inhibitory effect on ER+ and HER2+ breast cancer cells compared to petroleum ether or N-butanol extracts. APR inhibited ER+ and HER2+ breast cancer cell growth, proliferation, migration, and invasion via the phosphatidylinositol-3 kinase (PI3K)/AKT pathway. CONCLUSIONS: APR had a significant inhibitory effect on ER+ and HER2+ breast cancer cells via the PI3K/AKT signaling pathway. Therefore, APR may be useful for preventing ER+ and HER2+ breast tumor growth, proliferation, migration, and invasion.

The preventive and therapeutic effects of probiotics on mastitis: A systematic review and meta-analysis.

Acute mastitis is one of the main reasons why breastfeeding women stop breastfeeding, and medication should be used with caution. Considering the uncertainty of mastitis infection and the indications of antibiotic use, as well as the problem of drug resistance and the safety of medication during lactation, probiotics have become an alternative treatment choice. However, a meta-analysis of the effects of probiotics in preventing and treating lactational mastitis is still lacking. Therefore, we searched six electronic databases and the sites of clinical trial registration, a total of six randomized controlled trials were included in this meta-analysis, which showed that oral probiotics during pregnancy can reduce the incidence of mastitis (RR: 0.49, 95% CI: 0.35 to 0.69; p<0.0001). After oral administration of probiotics, the counts of bacteria in the milk of healthy people and mastitis patients were both significantly reduced (in healthy people: MD: -0.19, 95% CI: -0.23 to -0.16, p<0.00001; in mastitis patients: MD: -0.89, 95% CI: -1.34 to -0.43, p = 0.0001). These indicate that to a certain extent, probiotics are beneficial in reducing the incidence rate of mastitis during lactation and some related mastitis symptoms. However, high-quality multicenter clinical trials are still needed to support this result.

Efficacy and safety of microwave ablation for benign breast lesions: a systematic review and meta-analysis.

Aim: We performed a systematic review and meta-analysis to evaluate the efficacy and safety of microwave ablation (MWA) for benign breast lesions. Material and methods: PubMed, Embase, Web of Science, Cochrane Library databases, China National Knowledge Infrastructure, and Wanfang Data Knowledge Service Platform databases were searched. Results: A total of 10 studies were included, giving a sample size of 1241 patients and 2729 benign breast lesions. The first complete ablation success rate was 96%. The volume reduction ratio (VRR) after 3/6/12 months was 47.4%, 62.1%, and 85.8%, respectively. After 12 months, the lesion disappearance rate was 53.6%, and the efficiency rate was 99%. The rate of excellent cosmesis was 88% and the rate of good cosmesis was 10%. The complication rate was 2%. Conclusions: MWA is safe and effective for treating benign breast lesions. It can be a promising minimally invasive choice for benign breast lesions.

Transmissibility of tuberculosis among students and non-students: an occupational-specific mathematical modelling.

BACKGROUND: Recently, despite the steady decline in the tuberculosis (TB) epidemic globally, school TB outbreaks have been frequently reported in China. This study aimed to quantify the transmissibility of Mycobacterium tuberculosis (MTB) among students and non-students using a mathematical model to determine characteristics of TB transmission. METHODS: We constructed a dataset of reported TB cases from four regions (Jilin Province, Xiamen City, Chuxiong Prefecture, and Wuhan City) in China from 2005 to 2019. We classified the population and the reported cases under student and non-student groups, and developed two mathematical models [nonseasonal model (Model A) and seasonal model (Model B)] based on the natural history and transmission features of TB. The effective reproduction number (Reff) of TB between groups were calculated using the collected data. RESULTS: During the study period, data on 456,423 TB cases were collected from four regions: students accounted for 6.1% of cases. The goodness-of-fit analysis showed that Model A had a better fitting effect (P < 0.001). The average Reff of TB estimated from Model A was 1.68 [interquartile range (IQR): 1.20-1.96] in Chuxiong Prefecture, 1.67 (IQR: 1.40-1.93) in Xiamen City, 1.75 (IQR: 1.37-2.02) in Jilin Province, and 1.79 (IQR: 1.56-2.02) in Wuhan City. The average Reff of TB in the non-student population was 23.30 times (1.65/0.07) higher than that in the student population. CONCLUSIONS: The transmissibility of MTB remains high in the non-student population of the areas studied, which is still dominant in the spread of TB. TB transmissibility from the non-student-to-student-population had a strong influence on students. Specific interventions, such as TB screening, should be applied rigorously to control and to prevent TB transmission among students.

Study on the interaction between different pathogens of Hand, foot and mouth disease in five regions of China.

Objectives: This study aims to explore the interaction of different pathogens in Hand, foot and mouth disease (HFMD) by using a mathematical epidemiological model and the reported data in five regions of China. Methods: A cross-regional dataset of reported HFMD cases was built from four provinces (Fujian Province, Jiangsu province, Hunan Province, and Jilin Province) and one municipality (Chongqing Municipality) in China. The subtypes of the pathogens of HFMD, including Coxsackievirus A16 (CV-A16), enteroviruses A71 (EV-A71), and other enteroviruses (Others), were included in the data. A mathematical model was developed to fit the data. The effective reproduction number (R eff ) was calculated to quantify the transmissibility of the pathogens. Results: In total, 3,336,482 HFMD cases were collected in the five regions. In Fujian Province, the R eff between CV-A16 and EV-A71&CV-A16, and between CV-A16 and CV-A16&Others showed statistically significant differences (P < 0.05). In Jiangsu Province, there was a significant difference in R eff (P < 0.05) between the CV-A16 and Total. In Hunan Province, the R eff between CV-A16 and EV-A71&CV-A16, between CV-A16 and Total were significant (P < 0.05). In Chongqing Municipality, we found significant differences of the R eff (P < 0.05) between CV-A16 and CV-A16&Others, and between Others and CV-A16&Others. In Jilin Province, significant differences of the R eff (P < 0.05) were found between EV-A71 and Total, and between Others and Total. Conclusion: The major pathogens of HFMD have changed annually, and the incidence of HFMD caused by others and CV-A16 has surpassed that of EV-A71 in recent years. Cross-regional differences were observed in the interactions between the pathogens.

The signaling pathways and targets of traditional Chinese medicine and natural medicine in triple-negative breast cancer.

ETHNOPHARMACOLOGICAL RELEVANCE: Triple-negative breast cancer (TNBC) has a poorer prognosis than other subtypes due to its strong invasion and higher risk of distant metastasis. Traditional Chinese medicine (TCM) and natural medicine have the unique advantages of multitargets and small side-effects and may be used as long-term complementary and alternative therapies. AIM OF THE REVIEW: The present article summarizes the classical signaling pathways and potential targets by the action of TCM and natural medicine (including extracts, active constituents and formulas) on TNBC and provides evidence for its clinical efficacy. METHODS: The literature information was acquired from the scientific databases PubMed, Web of Science and CNKI from January 2010 to June 2020, and it was designed to elucidate the internal mechanism and role of TCM and natural medicine in the treatment of TNBC. The search key words included "Triple negative breast cancer" or "triple negative breast carcinoma", "TNBC" and "traditional Chinese medicine" or "Chinese herbal medicine", "medicinal plant", "natural plant", and "herb". RESULTS: We described the antitumor activity of TCM and natural medicine in TNBC based on different signaling pathways. Plant medicine and herbal formulas regulated the related gene and protein expression via pathways such as PI3K/AKT/mTOR, MAPK and Wnt/ß-catenin, which inhibit the growth, proliferation, migration, invasion and metastasis of TNBC cells. CONCLUSION: The inhibitory effect of TCM and natural medicine on tumors was reflected in multiple levels and multiple pathways, providing reasonable evidence for new drug development. To make TCM and natural medicine widely and flexibly used in clinical practice, the efficacy, safety and mechanism of action need more in-depth experimental research.

Epidemiological Characteristics and Transmissibility for SARS-CoV-2 of Population Level and Cluster Level in a Chinese City.

Background: To date, there is a lack of sufficient evidence on the type of clusters in which severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is most likely to spread. Notably, the differences between cluster-level and population-level outbreaks in epidemiological characteristics and transmissibility remain unclear. Identifying the characteristics of these two levels, including epidemiology and transmission dynamics, allows us to develop better surveillance and control strategies following the current removal of suppression measures in China. Methods: We described the epidemiological characteristics of SARS-CoV-2 and calculated its transmissibility by taking a Chinese city as an example. We used descriptive analysis to characterize epidemiological features for coronavirus disease 2019 (COVID-19) incidence database from 1 Jan 2020 to 2 March 2020 in Chaoyang District, Beijing City, China. The susceptible-exposed-infected-asymptomatic-recovered (SEIAR) model was fitted with the dataset, and the effective reproduction number (Reff ) was calculated as the transmissibility of a single population. Also, the basic reproduction number (R0) was calculated by definition for three clusters, such as household, factory and community, as the transmissibility of subgroups. Results: The epidemic curve in Chaoyang District was divided into three stages. We included nine clusters (subgroups), which comprised of seven household-level and one factory-level and one community-level cluster, with sizes ranging from 2 to 17 cases. For the nine clusters, the median incubation period was 17.0 days [Interquartile range (IQR): 8.4-24.0 days (d)], and the average interval between date of onset (report date) and diagnosis date was 1.9 d (IQR: 1.7 to 6.4 d). At the population level, the transmissibility of the virus was high in the early stage of the epidemic (Reff = 4.81). The transmissibility was higher in factory-level clusters (R0 = 16) than in community-level clusters (R0 = 3), and household-level clusters (R0 = 1). Conclusions: In Chaoyang District, the epidemiological features of SARS-CoV-2 showed multi-stage pattern. Many clusters were reported to occur indoors, mostly from households and factories, and few from the community. The risk of transmission varies by setting, with indoor settings being more severe than outdoor settings. Reported household clusters were the predominant type, but the population size of the different types of clusters limited transmission. The transmissibility of SARS-CoV-2 was different between a single population and its subgroups, with cluster-level transmissibility higher than population-level transmissibility.

Feasibility of Booster Vaccination in High-Risk Populations for Controlling Coronavirus Variants - China, 2021.

INTRODUCTION: Vaccination booster shots are completely necessary for controlling breakthrough infections of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in China. The study aims to estimate effectiveness of booster vaccines for high-risk populations (HRPs). METHODS: A vaccinated Susceptible-Exposed-Symptomatic-Asymptomatic-Recovered/Removed (SEIAR) model was developed to simulate scenarios of effective reproduction number (R eff ) from 4 to 6. Total number of infectious and asymptomatic cases were used to evaluated vaccination effectiveness. RESULTS: Our model showed that we could not prevent outbreaks when covering 80% of HRPs with booster unless R eff =4.0 or the booster vaccine had efficacy against infectivity and susceptibility of more than 90%. The results were consistent when the outcome index was confirmed cases or asymptomatic cases. CONCLUSIONS: An ideal coronavirus disease 2019 (COVID-19) booster vaccination strategy for HRPs would be expected to reach the initial goal to control the transmission of the Delta variant in China. Accordingly, the recommendation for the COVID-19 booster vaccine should be implemented in HRPs who are already vaccinated and could prevent transmission to other groups.

Odd-numbered agaro-oligosaccharides alleviate type 2 diabetes mellitus and related colonic microbiota dysbiosis in mice.

Agaro- and neoagaro-oligosaccharides with even-numbered sugar units possess a variety of biological activities. However, the effects of the odd-numbered oligosaccharides from Gracilaria agarose (OGAOs) on type 2 diabetes mellitus (T2DM) have not been reported. In this study, we aimed to evaluate the effects of OGAOs on anti-T2DM from different aspects. We found that OGAOs treatment could alleviate oxidative stress, inflammation, and the related hyperglycemia, insulin resistance, lipid accumulation, and obesity in high-fat diet (HFD) induced T2DM. Investigation of the underlying mechanism showed that colitis and colonic microbiota dysbiosis in T2DM mice were ameliorated after OGAOs treatment. First, OGAOs increased the expression of ZO-1, occludin, and AMPK, and suppressed the TLR4/MAPK/NF-κB pathway in colon indicating that OGAOs enhance intestinal integrity and conduct the anti-apoptosis effects to prevent the invasion of toxins and harmful microorganisms. Moreover, the relative abundance of Akkermansia was significantly upregulated in the gut microbiome of T2DM mice associated with a dramatic decrease of the relative abundance of Helicobacter, which are both beneficial for alleviating colitis and T2DM. In addition, Spearman's correlation analysis indicated that changes in the colonic microbiota could regulate oxidative stress, inflammation, and hyperlipidemia. In summary, the underlying mechanism of OGAOs on alleviating colitis and colonic microbiota dysbiosis in T2DM has been intensively studied, illustrating that OGAOs could be further developed as a potential pharmaceutical agent for T2DM.