A human pluripotent stem cell-based model of SARS-CoV-2 infection reveals an ACE2-independent inflammatory activation of vascular endothelial cells through TLR4.

To date, the direct causative mechanism of SARS-CoV-2-induced endotheliitis remains unclear. Here, we report that human ECs barely express surface ACE2, and ECs express less intracellular ACE2 than non-ECs of the lungs. We ectopically expressed ACE2 in hESC-ECs to model SARS-CoV-2 infection. ACE2-deficient ECs are resistant to the infection but are more activated than ACE2-expressing ones. The virus directly induces endothelial activation by increasing monocyte adhesion, NO production, and enhanced phosphorylation of p38 mitogen-associated protein kinase (MAPK), NF-κB, and eNOS in ACE2-expressing and -deficient ECs. ACE2-deficient ECs respond to SARS-CoV-2 through TLR4 as treatment with its antagonist inhibits p38 MAPK/NF-κB/ interleukin-1ß (IL-1ß) activation after viral exposure. Genome-wide, single-cell RNA-seq analyses further confirm activation of the TLR4/MAPK14/RELA/IL-1ß axis in circulating ECs of mild and severe COVID-19 patients. Circulating ECs could serve as biomarkers for indicating patients with endotheliitis. Together, our findings support a direct role for SARS-CoV-2 in mediating endothelial inflammation in an ACE2-dependent or -independent manner.

Clinical research on minimally invasive rotary resection in the treatment of lactational breast abscess.

BACKGROUND: This study sought to compare the efficacy of minimally invasive rotatory resection plus catheter drainage and puncture abscess aspiration in the treatment of lactational breast abscess. METHODS: The data of 80 patients with lactational breast abscess at the Department of Mammary Gland, Tianjin Central Hospital of Gynecology Obstetrics, from January 2019 to January 2020 were retrospectively collected. The patients were divided into the minimally invasive group (n=30) and the abscess aspiration group (n=50). Clinical data, including age, parity, preoperative body temperature, postpartum time, time of abscess cavity generation, diameter and position of abscess cavity, intraoperative drainage, bleeding, recovery time, and satisfaction score, were collected and compared between the 2 groups of patients. RESULTS: The baseline data between the 2 groups were not significantly different. The minimally invasive group had a higher single drainage volume and single bleeding volume (P<0.05), and lower cumulative pain score (P<0.05); the cumulative bleeding volume did not differ significantly between the 2 groups (P>0.05). For the abscess cavities with a diameter ≤5 cm, the single drainage volume, single bleeding volume, and single pain score in the minimally invasive group were significantly higher than those in the abscess aspiration group (P<0.05); the total drainage volume was similar between the 2 groups (P>0.05). For the abscess cavities with a diameter >5 cm, the single drainage volume and single bleeding volume in the minimally invasive group were significantly higher than those in the abscess aspiration group (P<0.05), while the cumulative bleeding volume, single pain score, and cumulative pain score were lower in the minimally invasive group (P<0.05). There was no significant difference in total drainage between the 2 groups (P>0.05). CONCLUSIONS: When the abscess cavity is ≤5 cm in diameter and the abscess is superficial, puncture and abscess aspiration may be more advantageous than the minimally invasive rotatory resection plus catheter drainage. When the abscess cavity is >5 cm in diameter and there are multiple abscesses or deep large abscess, the efficacy of minimally invasive rotary resection plus catheter drainage may be better than the puncture and abscess aspiration.

Endothelial Agrin Is Dispensable for Normal and Tumor Angiogenesis.

Recently, the extracellular matrix protein agrin has been reported to promote tumor angiogenesis that supports tumorigenesis and metastasis; however, there is a lack of in vivo genetic evidence to prove whether agrin derived from the tumors or endothelial cells (ECs) systemically should be the therapeutic target. To date, the physiological role of endothelial agrin has also not been investigated. In the EC-specific agrin knockout mice, we observed normal endothelial and haematopoietic cell development during embryogenesis. Moreover, these mice develop normal vascular barrier integrity and vasoreactivity at the adult stage. Importantly, the growth of localized or metastatic cancer cells was not affected after implantation into endothelial agrin depleted mice. Mechanistically, agrin did not regulate endothelial ERK1/2, YAP or p53 activation in vivo that is central to support endothelial proliferation, survival and invasion. Cumulatively, our findings may suggest that agrin could play a redundant role in endothelial development during physiological and tumor angiogenesis. Targeting the endothelial derived agrin might not be effective in inhibiting tumor angiogenesis.

Molecular Identification Based on Chloroplast Sequences and Anti-complementary Activity Comparison of Juniperus Samples from the Qinghai-Tibet Plateau.

Juniperus (Cupressaceae, Pinales) plants are widely distributed in the Qinghai-Tibet Plateau of China. The leaves and twigs of at least 8 Juniperus species (J. pingii, J. pingii var. wilsonii, J. squamata, J. recurva var. coxii, J. saltuaria, J. indica, J. tibetica and J. convallium var. microsperma) have been used as the Tibetan medicine Xuba. At present, it is difficult to distinguish among the original species of Xuba based only on their similar morphological characteristics. However, in our previous studies, 4 Xuba samples from different Juniperus species exhibited significant differences in both anticomplementary activity in vitro and anti-inflammatory effects on acute lung injury in vivo. To identify the effective original species of Xuba reliably, in this study, we developed a sequencing-based DNA molecular technology to distinguish 14 populations of 8 Juniperus species collected from Tibet region, using trnS-G, trnD - T, and petN-psbM genomic regions to build phylogenetic trees. In addition, their anticomplementary activities were evaluated. The results showed that combined sequence of these 3 genomic regions could identify 8 Juniperus species clearly and clustered individuals of one species but from different locations, whichever phylogenetic tree was constructed. Moreover, the anticomplementary activities of the 8 species were clustered into 2 groups. Among them, J. saltuaria and J. recurva var. coxii, which formed an independent branch apart from the other 6 species in phylogenetic trees, were the most potent (CH50: 0.029 - 0.032 mg/mL). Consequently, DNA identification of Juniperus using the combined sequence could provide beneficial guidance for further efficacy evaluation and quality control of Xuba.

Beneficial effects on H1N1-induced acute lung injury and structure characterization of anti-complementary acidic polysaccharides from Juniperus pingii var. wilsonii.

Juniperus pingii var. wilsonii has been traditionally used in Tibetan medicine for the treatment of inflammatory diseases. In the present study, J. pingii var. wilsonii polysaccharides (JPWP), with high content of d­galacturonic acid, showed potent anti-complementary activity in vitro and significantly attenuated acute lung injury (ALI) induced by H1N1 influenza virus in vivo through reducing the inflammatory responses, alleviating oxidative stress and inhibiting the activation of complement. Thus, anti-complementary activity-guided fractionation of JPWP led to the isolation of an acidic homogeneous polysaccharide, JPWP-PS, whose structure was further elucidated by acid hydrolysis, PMP derivation, methylation and NMR analysis. JPWP-PS had potent anti-complementary activity with the CH50 value of 0.073 ±â€¯0.009 mg/mL, and was characterized by the residues of T-Araf-(1→, →3)-Araf-(1→, →3,5)-Araf-(1→, →3)-Galp-(1→ and →4)-GalpA-(1→.

Anti-complementary constituents of Anchusa italica.

Activity-guided fractionation for complement inhibitors led to the isolation of 24 known compounds from Anchusa italica. Chemical types include eight megastigmane compounds, five triterpenoid compounds, five lignan compounds, three flavonoid compounds, two alkaloid compounds and one phenthyl alcohol compound. Among which, a lignan (medioresinol), an alkaloid (5-hydroxypyrrolidin-2-one) and a flavonoid (5-hydroxyl-3', 4', 6, 7-tetramethoxy flavone) exhibited better anticomplementary effects with CH50 values ranging from 0.07 to 0.82 mM, which are plausible candidates for developing potent anticomplementary agents.

Anti-complementary constituents of Viola kunawarensis.

Activity-guided fractionation for complement inhibitors led to the isolation of 22 known compounds from Viola kunawarensis. Chemical types include six sterol compounds, three coumarin compounds, five megastigmane compounds, two triterpenoid compounds, two phenylpropanoid compounds, one chlorophyll, one amide, and two lipid compounds. Among which, two sterols (stigmasta-4-ene-3ß,6ß-diol and saringosterone), one amide (aurantiamide acetate) and a norsesquiterpenoid (solalyratin B) exhibited better anti-complementary effects with CH50 values ranging from 0.02 to 0.08 mM, which are plausible candidates for developing potent anti-complementary agents.

Transforming growth factor ß-activated kinase 1 inhibitor suppresses the proliferation in triple-negative breast cancer through TGF-ß/TGFR pathway.

Breast cancer is one of the most invasive cancer types in female population. The functional activity of Transforming growth factor ß-activated kinase 1 (TAK1) in breast cancer progression increasingly attracts attention as it provides a potential target for antibreast cancer drug development. However, the fundamental role of TAK1 for triple-negative breast cancer (TNBC) progression and the effect of potential anti-TAK1 drug candidate needs to be further evaluated. Herein, we focused on the role of TAK1 in human breast cancer cells, and we hypothesized that the inhibition of TAK1 activation can repress the growth of human TNBC cells. We found that the TAK1 is robustly activated within cancer cell population of clinic-derived TNBC samples and the human breast cancer cell lines in culture. Furthermore, we determined the effect of 5Z-7-oxozeaenol (5Z-O), a TAK1-specific small molecule inhibitor, on proliferation of human TNBC cell line. 5Z-O treatment significantly suppressed the proliferation of human TNBC cells. Collectively, these demonstrate the role of TAK1 in human breast cancer and the antiproliferate effect of TAK1 inhibitor. Our study sets the stage for further research on TAK1 as a promising target for development of anti-TNBC drugs and therapeutic strategies.