Isobavachalcone Induces Multiple Cell Death in Human Triple-Negative Breast Cancer MDA-MB-231 Cells.

Standardized treatment guidelines and effective drugs are not available for human triple-negative breast cancer (TNBC). Many efforts have recently been exerted to investigate the efficacy of natural compounds as anticancer agents owing to their low toxicity. However, no study has examined the effects of isobavachalcone (IBC) on the programmed cell death (PCD) of human triple-negative breast MDA-MB-231 cancer cells. In this study, IBC substantially inhibited the proliferation of MDA-MB-231 cells in concentration- and time-dependent manners. In addition, we found that IBC induced multiple cell death processes, such as apoptosis, necroptosis, and autophagy in MDA-MB-231 cells. The initial mechanism of IBC-mediated cell death in MDA-MB-231 cells involves the downregulation of Akt and p-Akt-473, an increase in the Bax/Bcl-2 ratio, and cleaved caspases-3 induced apoptosis; the upregulation of RIP3, p-RIP3 and MLKL induced necroptosis; as well as a simultaneous increase in LC3-II/I ratio induced autophagy. In addition, we observed that IBC induced mitochondrial dysfunction, thereby decreasing cellular ATP levels and increasing reactive oxygen species accumulation to induce PCD. These results suggest that IBC is a promising lead compound with anti-TNBC activity.

Epididymal initial segment-specific Cre recombinase activity in Lcn8-Cre knock-in mice.

BACKGROUND: Sperm acquire the ability to fertilize ova through a complex process of epididymal maturation. To identify the functions of genes expressed in the proximal epididymis, mouse models specific to this region are needed. METHODS AND RESULTS: A Lcn8-Cre knock-in mouse line was generated using CRISPR/Cas9 technology. A 37 bp coding sequence of Lcn8 from the ATG start codon was replaced by an NLS-Cre-polyA cassette, resulting in Cre expression and the absence of Lcn8. Epididymal initial segment-specific Cre expression was identified using RT-PCR and western blotting, and the spatial-temporal Cre activity was further confirmed by using the Rosa26tdTomato reporter mice. Immunofluorescence staining showed that active Cre recombinase was present in the principal cells. Histological analyses of sperm and epididymides, and the four-month mating tests, were used to confirm that Cre expression did not affect normal development and male fecundity. CONCLUSIONS: The novel Lcn8-Cre mice can be used to establish epididymal initial segment-specific conditional knock-out mouse models.

A novel mouse line with epididymal initial segment-specific expression of Cre recombinase driven by the endogenous Lcn9 promoter.

Spermatozoa released from testes undergo a maturation process and acquire the capacity to fertilize ova through epididymal transit. The epididymis is divided into four regions, each with unique morphology, gene profile, luminal microenvironment and distinct function. To study the functions of relevant genes in the epididymal initial segment (IS), a novel IS-specific mouse model, Lcn9-Cre knock-in (KI) mouse line was generated via CRISPR/Cas9 technology. The TAG stop codon was replaced by a 2A-NLS-Cre cassette, resulting in the co-expression of Lcn9 and Cre recombinase. IS-specific Cre expression was first observed from postnatal day 17. Using the Rosa26tdTomato reporter mice, the Cre-mediated DNA recombination was detected exclusively in principal cells. The epididymal IS-specific Cre activity in vivo was further confirmed using Lcn9-Cre mice crossed with a mouse strain carrying Tsc1 floxed alleles (Tsc1flox/+). Cre expression did not affect either normal development or male fecundity. Different from any epididymis-specific Cre mice reported previously, the novel Lcn9-Cre mouse line can be used to introduce entire IS-specific conditional gene editing for gene functional study.

[Clinical Epidemiological Characteristics of Newly Reported HIV/AIDS in a Certain General Hospital from 2001 to 2017].

OBJECTIVE: To determine the clinical epidemiological characteristics of newly reported human immunodeficiency virus/acquired immunodeficiency syndrome （HIV/AIDS）in southwestern China from 2001 to 2017. METHODS: Clinical data of newly diagnosed HIV/AIDS from 2001 to 2017 in the West China Hospital of Sichuan University were reviewed and analyze. RESULTS: A total of 1 520 228 patients were screened for HIV, including 285 983 outpatient and emergency patients and 1 234 245 inpatients. About 4 037 (0.27%) patients were confirmed with HIV/AIDS. The confirmation rate increased from 2001 to 2013, followed by a slight decline from 2014 to 2017. The male to female sex ratio of confirmed HIV/AIDS was 3.49:1 from 2001 to 2017, ranging from 1.65:1 to 5.08:1. The majority of patients were identified as Han (88.23%), had low education (58.66%), and married (54.75%). Peasants/herdsman comprised 26.33% of the patients. The proportion of young (15-29 years old), and middle-aged (≥50 years old) patients and those who were unmarried and had high education (senior high school and above) increased over time. Heterosexual transmission remained stable at about 60% while homosexual transmission increased by about 15% ( χ 2=14.436, P＜0.005) since 2008. Transmissions through drug abuse( χ 2=71.633, P＜0.005) and blood( χ 2=16.672, P＜0.005) decreased. Of the 899 female newly reported HIV/ADIS patients, 77.20% were infected through heterosexual relationship. In comparison, of the 3 138 male patients, 61.41% were infected through heterosexual and 18.10% through homosexual relationships. Homosexual transmissions decreased with age, but heterosexual transmissions increased with age. Mother-to-child transmissions were concentrated in those between 0 and 15 years old (100%). CONCLUSION: Newly diagnosed HIV/AIDS cases increased over the years in the West China Hospital of Sichuan University, in particular in those of young and middle-aged, highly educated and unmarried. Heterosexual transmissions remain the main route.

Five-Golden-Flowers Tea: Green Extraction and Hepatoprotective Effect against Oxidative Damage.

The consumption of herbal teas has become popular in recent years due to their attractive flavors and outstanding antioxidant properties. The Five-Golden-Flowers tea is a herbal tea consisting of five famous edible flowers. The effects of microwave-assisted extraction parameters on the antioxidant activity of Five-Golden-Flowers tea were studied by single-factor experiments, and further investigated using response surface methodology. Under the optimal parameters (53.04 mL/g of solvent/material ratio, 65.52 °C, 30.89 min, and 500 W), the ferric-reducing antioxidant power, Trolox equivalent antioxidant capacity, and total phenolic content of the herbal tea were 862.90 ± 2.44 µmol Fe2+/g dry weight (DW), 474.37 ± 1.92 µmol Trolox/g DW, and 65.50 ± 1.26 mg gallic acid equivalent (GAE)/g DW, respectively. The in vivo antioxidant activity of the herbal tea was evaluated on alcohol-induced acute liver injury in mice. The herbal tea significantly decreased the levels of aspartate aminotransferase, total bilirubin, and malonaldehyde at different doses (200, 400, and 800 mg/kg); improved the levels of liver index, serum triacylglycerol, and catalase at dose of 800 mg/kg. These results indicated its role in alleviating hepatic oxidative injury. Besides, rutin, chlorogenic acid, epicatechin, gallic acid, and p-coumaric acid were identified and quantified by high performance liquid chromatography (HPLC), which could contribute to the antioxidant activity of the herbal tea.

Cation-dependent mannose-6-phosphate receptor functions as a pattern recognition receptor in anti-bacterial immunity of Marsupenaeus japonicus.

The cation-dependent mannose-6-phosphate receptor (CD-MPR) is a member of the P-type lectin family. As a type I transmembrane glycoprotein, it functions in the delivery of newly synthesized acid hydrolases from the trans-Golgi network to endosomes for their subsequent transfer to the lysosome by binding the mannose-6-phosphate receptor-recognition moieties in the hydrolases. However, the functions of CD-MPR in immune responses are seldom reported. In the present study, we identified a CD-MPR-like molecule in Marsupenaeus japonicus and designed it as MjCD-MPR. It was significantly upregulated after challenge with Vibrio anguillarum at the mRNA and protein levels. Knockdown of MjCD-MPR resulted in a significant increase in the amount of V. anguillarum in the hemolymph of shrimp, which suggested that MjCD-MPR plays a role in shrimp antibacterial defense. The recombinant extracytoplasmic region of MjCD-MPR could bind gram-positive and gram-negative bacteria by interaction with peptidoglycan, lipopolysaccharide, and lipoteichoic acid. MjCD-MPR showed no direct bacteriostatic or bacteriocidal activity. Knockdown of MjCD-MPR decreased the expression levels of several antimicrobial peptides (Alf-C1, Alf-E1, Crustin I-2, and Crustin I-3), suggesting that MjCD-MPR promotes the expression of antimicrobial peptides in shrimp. In summary, working as a pattern recognition receptor, MjCD-MPR recognizes invading bacteria and triggers the expression of AMPs against bacterial infection in shrimp.

Interaction of the Small GTPase Cdc42 with Arginine Kinase Restricts White Spot Syndrome Virus in Shrimp.

Many types of small GTPases are widely expressed in eukaryotes and have different functions. As a crucial member of the Rho GTPase family, Cdc42 serves a number of functions, such as regulating cell growth, migration, and cell movement. Several RNA viruses employ Cdc42-hijacking tactics in their target cell entry processes. However, the function of Cdc42 in shrimp antiviral immunity is not clear. In this study, we identified a Cdc42 protein in the kuruma shrimp (Marsupenaeus japonicus) and named it MjCdc42. MjCdc42 was upregulated in shrimp challenged by white spot syndrome virus (WSSV). The knockdown of MjCdc42 and injection of Cdc42 inhibitors increased the proliferation of WSSV. Further experiments determined that MjCdc42 interacted with an arginine kinase (MjAK). By analyzing the binding activity and enzyme activity of MjAK and its mutant, ΔMjAK, we found that MjAK could enhance the replication of WSSV in shrimp. MjAK interacted with the envelope protein VP26 of WSSV. An inhibitor of AK activity, quercetin, could impair the function of MjAK in WSSV replication. Further study demonstrated that the binding of MjCdc42 and MjAK depends on Cys271 of MjAK and suppresses the WSSV replication-promoting effect of MjAK. By interacting with the active site of MjAK and suppressing its enzyme activity, MjCdc42 inhibits WSSV replication in shrimp. Our results demonstrate a new function of Cdc42 in the cellular defense against viral infection in addition to the regulation of actin and phagocytosis, which has been reported in previous studies. IMPORTANCE The interaction of Cdc42 with arginine kinase plays a crucial role in the host defense against WSSV infection. This study identifies a new mechanism of Cdc42 in innate immunity and enriches the knowledge of the antiviral innate immunity of invertebrates.

Association Between Serum Uric Acid and Prevalence of Type 2 Diabetes Diagnosed using HbA1c Criteria Among Chinese Adults in Qingdao, China.

OBJECTIVE: To determine whether elevated serum uric acid (UA) levels are associated with type 2 diabetes diagnosed using HbA1c levels among Chinese adults. METHODS: We conducted two population-based cross-sectional studies in Qingdao in China in 2006 and 2009. A total of 6894 (39.4% men) subjects aged 35-74 years were included in the data analysis. Newly diagnosed diabetes was defined as HbA1c level of ⋝6.5%, and prediabetes was classified as HbA1c level between 5.7% and 6.4% according to the International Diabetes Federation criteria. Multivariate logistic regression was employed to assess the association between UA and prevalence of type 2 diabetes defined using Glycated hemoglobin A1c (HbA1c) levels. RESULTS: Subjects with prediabetes had higher UA levels than those with normal glucose tolerance, newly diagnosed diabetes, and known diabetes, with corresponding values of 325.1 (82.5) µmol/L, 310.9 (84.2) µmol/L, 291.3 (81.7) µmol/L, 305.2 (83.6) µmol/L, respectively (P<0.001 for all comparisons). Binary logistic regression analysis showed that UA was a possible predictor for the prevalence of type 2 diabetes diagnosed using HbA1c levels, and the second quartile of UA levels had a higher odds ratio (OR: 4.088; 95% CI: 2.900-5.765) for HbA1c than the other quartiles after adjusting for age, body mass index, sex, marital status, education, income, alcohol consumption, smoking, and cardiometabolic parameters. CONCLUSION: Serum UA is significantly associated with type 2 diabetes diagnosed using HbA1c levels, independent of other cardiometabolic parameters.

Effect of superoxide dismutase and malondialdehyde metabolic changes on carcinogenesis of gastric carcinoma.

AIM: To investigate the relationship between the superoxide dismutase (SOD), malondialdehyde (MDA) metabolic changes and the gastric carcinogenesis. METHODS: The SOD activity and MDA content were measured in the gastric tissues from the focus center, peripheral and far-end areas of gastric carcinoma (n = 52) and gastric ulcer (n = 10). All the tissues were subjected to routine histological examinations and classifications. RESULTS: The SOD activity was greatly reduced but the MDA content was markedly increased in the center areas of the non-mucous gastric carcinoma (non-MGC); and the poorly differentiated gastric carcinoma varied. The SOD activity was gradually decreased and the MDA content was gradually increased in the tissues from the focus far-end, peripheral to center areas of non-MGC. Both of the SOD activity and the MDA content were significantly declined and were respectively at same low level in the tissues from the focus center, peripheral, and far-end area with the mucous gastric carcinoma (MGC). In contrast to the gastric ulcer and grade I or II of non-MGC, the same level of the SOD activity and the MDA content were found in the focus center areas. Between non-MGC (groups A-D) and gastric ulcer (group F), the differences of SOD activity and MDA content were very noticeable in the gastric tissues from the focus peripheral and far-end areas, in which the SOD activity showed noticeable increase and the MDA content showed noticeable decrease in the gastric ulcer. CONCLUSION: The active free radical reaction in the gastric tissues can induce the carcinogenesis of non-MGC. The utmost low ability of antioxidation in the gastric tissues can induce the carcinogenesis of MGC. The metabolic change of the free radicals centralized mostly in the center of ulcerated lesions only, which suggested the ability of antioxidation was declined only in these lesions. However, the metabolism of free radicals varied significantly and the ability of antioxidation declined not only in the local focus area but also in the abroad gastric tissues with gastric carcinoma.