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The coding sequences (CDS) of TFAM and TFB2M genes from Jiaxing Black Pig (JBP) were first obtained by RT-PCR and DNA-seq in the present study. Sequence analyses showed that the TFAM gene contains a 741-bp CDS region encoding 246 amino acids sharing a 100% homology with the sequence on NCBI, while TFB2M gene contains a CDS region of 1176 bp encoding 391 amino acids with two missense mutations. The results of quantitative Real-Time PCR for TFAM and TFB2M revealed that transcripts of the genes were both presented at the highest levels in spleen tissue followed by liver tissue, while the least levels in longissimus dorsi muscle (LDM), and obviously the higher levels in two adipose tissues than those in LDM tissue (PTFAM and TFB2M mRNA in particular tissues such as liver and LDM with carcass and meat quality traits including marbling score, as well as the content of saturated fatty acid (SFA), in JBP were found.

T cells modified to co-express cytokine or other factors with chimeric antigen receptor (CAR) can induce substantial and persistent increases in antitumor capacity in vivo. However, the uncontrolled expression of cytokines or factors can lead to the overactivation of immune cells, causing severe adverse events such as cytokine release syndrome (CRS) and neurotoxicity by CAR T cells with excessive growth potential. Conventional promoters are unregulated, and their expression is unlimited in T cells. In this study, by connecting the cytomegalovirus (CMV) enhancer, core interferon gamma (IFN-γ) promoter, and a T-lymphotropic virus long terminal repeat sequence (TLTR), we constructed and screened the chimeric promoter CIFT, which was highly expressed in some cell lines secreting IFN-γ and silenced in others. We placed this promoter upstream of the anti-programmed cell death protein 1 (anti-PD-1) antibody gene, and this construct was co-transfected with the CAR construct into T cells. In vitro or in vivo, CAR T cells showed increased secretion of anti-PD-1 antibody under control of the chimeric promoter CIFT. pS-CIFT-αPD-1/CAR T also had similar or lower PD-1 expression, higher levels of T cell activation, more release of IFN-γ, and better antitumor activity specifically against mesothelin-positive and PD-1 ligand 1 (PD-L1)-positive cell lines. The chimeric promoter may be a promising strategy to manipulate the content of immune checkpoint inhibitors or other proteins in future therapeutic approaches for cancer treatment., Graphical Abstract, Chimeric antigen receptor (CAR) T cell excessive growth always leads to severe adverse reactions such as cytokine release syndrome (CRS) and neurotoxicity. The source of this issue is considered to be the unbridled expression of cytokines or immune checkpoint inhibitors, which is caused by ineligible chimeric promoters. In this study, Qian and colleagues developed a chimeric promoter that is able to regulate anti-PD-1 antibody expression in CAR T cells.

Pathological pain is a hyperexcitability disorder. Since the excitability of a neuron is set and controlled by a complement of ion channels it expresses, in order to understand and treat pain, we need to develop a mechanistic insight into the key ion channels controlling excitability within the mammalian pain pathways and how these ion channels are regulated and modulated in various physiological and pathophysiological settings. In this review, we will discuss the emerging data on the expression in pain pathways, functional role and modulation of a family of voltage‐gated K+ channels called ‘M channels’ (KCNQ, Kv7). M channels are increasingly recognized as important players in controlling pain signalling, especially within the peripheral somatosensory system. We will also discuss the therapeutic potential of M channels as analgesic drug targets. Linked Articles This article is part of a themed section on Recent Advances in Targeting Ion Channels to Treat Chronic Pain. To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v175.12/issuetoc/

BACKGROUND: YiqiHuoxue decoction (YHD) is frequently prescribed to prevent and treat cardiovascular diseases. YHD inhibits platelet aggregation, however the underlying mechanisms are unclear. METHODS: The in vitro and in vivo anti-platelet and antithrombotic effects of YHD and ethanol-precipitated YHD (EYHD) and underlying mechanisms were investigated. Forty-six Sprague-Dawley (SD) rats and 36 male Kunming mice were examined. Ten SD rats were used to assess the cytotoxicity of YHD and EYHD by releasing lactate dehydrogenase from treated platelets. The remaining 36 SD rats were divided into six groups (six per group), including control saline (5 mL/kg), aspirin (20 mg/kg), YHD low dosage (0.2 g/kg), YHD high dosage (2.0 g/kg), 75% EYHD low dosage (0.2 g/kg), and 75% EYHD high dosage (2.0 g/kg) groups to detect platelet aggregation; the 36 Kunming mice were divided into 6 groups to detect mesenteric arterial thrombosis induction. Thromboxane B2 (TXB2) levels were determined by enzyme immunoassay. RESULTS: YHD high dosage and 75% EYHD (low and high dosage) inhibited ADP-induced platelet aggregation. Moreover, collagen-induced platelet aggregation was significantly suppressed by YHD (high dosage), 75% EYHD (high dosage), and 75% EYHD (low dose). Rats given 75% EYHD (high dose) displayed a marked reduction in collagen-induced platelet aggregation at 2 h post-administration. YHD and EYHD markedly prolonged the onset of thrombosis causing loose attachment of the thrombus to the vascular endothelium, but bleeding and clotting times were not significantly changed. Finally, YHD and EYHD markedly reduced TXB2 levels. CONCLUSIONS: YHD and EYHD effectively inhibit platelet activation and thrombosis, presumably by suppressing TXB2.

Argonaute2 (AGO2) protein is the active part of RNA-induced silencing complex, cleaving the target mRNA strand complementary to their bound siRNA. An increasing number of miRNAs has been identified as essential to angiogenesis of hepatocellular carcinoma (HCC). In this study we investigated how AGO2 affected HCC angiogenesis. Human HCC cell lines HepG2, Hep3B, Huh7, SMMC-7721, Bel-7404, MHCC97-H and LM-3, and human umbilical vein endothelial cells (HUVEC) were tested. The expression of AGO2 in HCC cells was knocked down with siRNA and restored using recombinant adenovirus expressing Ago2. The levels of relevant mRNAs and proteins were examined using RT-PCR, Western blot and EILSA. Nude mice were implanted with Huh7 or SMMC-7721 cells, and tumor volumes were measured. After the mice were euthanized, the xenograft tumors were used for immunohistological analysis. In 6 HCC cell lines, AGO2 protein expression was significantly correlated with VEGF expression (r=+0.79), and with VEGF secretion (r=+0.852). Knockdown of Ago2 in Huh7 cells and SMMC-7721 cells substantially decreased VEGF expression, whereas the restoration of AGO2 reversed both VEGF expression and secretion. Furthermore, knockdown of Ago2 significantly up-regulated the expression of PTEN (a tumor suppressor involved in the inhibition of HCC angiogenesis), and vice versa. Moreover, the specific PTEN inhibitor bisperoxovanadate (7, 14, 28 nmol/L) dose-dependently restored the expression of VEGF and the capacity of HCC cells to induce HUVECs to form capillary tubule structures. In the xenograft nude mice, knockdown of Ago2 markedly suppressed the tumor growth and decreased PTEN expression and CD31-positive microvascular in the xenograft tumors. A direct relationship exists between the miRNA processing machinery AGO2 and HCC angiogenesis that is mediated by the AGO2/PTEN/VEGF signaling pathway. The results suggest the high value of Ago2 knockdown in anti-angiogenesis therapy for HCC.

Rat embryonic stem (ES) cells hold great interest for the research of neurodevelopment and neurodegenerative diseases. However, neural conversion of rat ES cells in vitro has proven to be a challenge owing to the proliferation arrest and apoptosis. Here we report that rat ES cells can commit efficiently to a neural fate in the presence of CHIR99021 and Y-27632 (CY medium). In addition, CHIR99021 is crucial for maintaining the metabolic activity of differentiated rat ES cells, while Y-27632 facilitates the neural differentiation of rat ES cells by inhibiting bone morphogenetic protein expression. The chemical-defined CY medium also provides a platform for exploring the mechanism of neural commitment and optimizing the production efficiency of neural precursor from rat ES cells.