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Hawthorn is a berry-like fruit from the species of Crataegus. In China, it has another more famous name, Shan-Zha, which has been used to improve digestion as a traditional Chinese medicine or food for thousands of years. Moreover, during the last decades, hawthorn has received more attention because of its potential to treat cardiovascular diseases. However, currently, only fruits of C. pinnatifida and C. pinnatifida var. major are included as Shan-Zha in the Chinese Pharmacopoeia. In this study, our results showed that the ethanol extract of Zhongtian hawthorn, a novel grafted cultivar of C. cuneata (wild Shan-Zha), could markedly reduce body weight and levels of serum total cholesterol, triglyceride, low-density lipoprotein cholesterol, and liver cholesterol of hyperlipidemia mice. It could suppress the stimulation effect of high-fat diet on the transcription of 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR) and p65, and counteract the downregulation of CYP7A1 and LDLR. In addition, the results of luciferase reporter assay and Western blot showed that the transcriptional activity of HMGCR promoter was inhibited by Zhongtian hawthorn ethanol extract in a dose-dependent manner, while overexpression of p65 could reverse this transcriptional repression effect. These results suggested that Zhongtian hawthorn could provide health benefits by counteracting the high-fat diet-induced hypercholesteolemic and hyperlipidemic effects in vivo, and the mechanism underlying this event was mainly dependent on the suppressive effect of Zhongtian hawthorn ethanol extract on the transcription of HMGCR via nuclear factor-kappa B (NF-κB) signal pathway. Therefore, this novel cultivar of hawthorn cultivar which has much bigger fruits, early bearing, high yield, cold resistance, and drought resistance, might be considered as a good alternative to Shan-Zha and has great value in the food and medicine industry. In addition, to our best knowledge, this is also the first report that the extract of Crataegus could suppress the transcription of HMGCR via NF-κB signal pathway.

The 3D configuration of graphene materials has been intensively investigated due to their novel properties in biomedical, electrical and optical applications. But few reports have been intentionally carried out to understand the biological influence of 3D graphene materials so far. Herein, we presented an evaluation of the in vitro cytotoxicity of 3D graphene sheets fabricated by carbonization of polydopamine (PDA) films on a template of aligned nanopore arrays (NPAs) on a stainless steel surface. The prepared 3D graphene sheets with a thickness of ∼20 nm displayed a nanoporous architecture that can be readily tuned by the NPA template to control the morphology of the 3D configuration. The in vitro toxicity of the nanoporous 3D graphene sheets with pore sizes of ∼50 nm and ∼240 nm was evaluated using NIH-3T3 fibroblasts as the representative mammal fibroblast cell type. The NPA structure exhibits enhanced properties in cell attachment, spreading, proliferation, and the assembly of focal adhesions and actin filament associated proteins. Morphology-dependent cytotoxicity aroused by the 3D graphene configuration should be attributed to the engulfment of the carbon nanospheres embedded in the 3D configuration and the lack of both focal adhesions and actin filament associated proteins assembling at the nanoscale.

Mutations of the Tp53 gene occur in a subset of patients with refractory/relapse B-cell lymphoma and confer an exceedingly adverse prognosis. Chimeric antigen receptor-modified (CAR) T-cell immunotherapy represents a novel promising treatment and has achieved impressive anti-tumor responses in patients with refractory or relapsed B cell malignancies. Whether CAR T cell therapy can improve the outcomes of refractory/relapse B-cell lymphoma with Tp53 mutations or high-risk functional classification of Tp53 mutations has not been fully investigated yet. In phase 2 trial, we evaluated the anti-CAR 19/22 cocktail therapy in patients with relapsed or refractory B-cell lymphoma with Tp53 mutations. The primary endpoint was the percentage of patients with an objective response (complete and partial response). The high-risk Tp53 mutations were defined as missense mutations, 'disruptive' mutations and the 'high-risk group of evolutionary action (EAp53) score. The outcome impact of CAR T cell therapy on high-risk Tp53 mutations were also assessed. Between September 2016 to January 2020, a total of 30 patients [median age, 44.0 years (range: 28.0 to 61.0)] were enrolled. The primary efficacy analysis showed that 76.7 % (95%CI: 61% to 93%) patients have best objective response(Figure 1A), while 60.0 % (95%CI: 41% to 79%) patients at month three maintained the objective response(Figure 1B). At a median follow-up of 7.95 months (range: 1.13 to 42.76), the 12-months progression-free survival and overall survival rate were 43.33% and 45.33% respectively(Figure 1C-D). The therapy led to similar serious and life-threatening toxic effects with those reported with other CAR T-cell therapies. For all 30 enrolled patients, total 32 Tp53 Mutations were detected in this cohort including 23 missense, 4 splice site mutations, 4 insertions, 2 deletions and 1 nonsense mutation. Two Tp53 mutations were found in 2 patients. 21 patients were assessed with Tp53 FISH and 14 patients (66.66%) had 17p-. There is no difference on ORR,OS and PFS in patients based on three different Tp53-specific scoring systems. The ORR at three months predicts the long-term survival of this group of patients (Figure 1E-F). CAR19/22 T-cell cocktail therapy can improve the outcome of r/r B-cell lymphoma with high-risk of functional classification of Tp53 mutation. The patients who achieved overall response at month three can greatly benefit from CAR19/22 T-cell cocktail therapy with long-term favorable outcome. Trial registration: ChiCTR-OPN-16008526. Key words: B cell lymphoma; CAR T-cell therapy; Tp53 mutation; outcome Figure 1 Disclosures No relevant conflicts of interest to declare.

High-dose chemotherapy followed by autologous stem cell transplantation (HDT-ASCT) is the standard of care for relapsed or refractory(R/R) B-cell lymphoma. Eligibility for HDT-ASCT is based upon the presence of chemo-sensitive disease to salvage therapy. Patients with a positive PET scan prior to ASCT have a substantially inferior suvival. Furthermore, the outcomes of patients who have progressive disease after salvage treatment is extremely dismal. We hypothesized that, following HSCT, the immunosuppressive microenvironment would be diminished as a result of myeloablative conditioning and that administering CAR-T cell therapy during the hematopoietic reconstitution could eradicate posttransplant residual disease, leading to lower relapse rates after HSCT. Based on our previous work that a dual-targeting CAR T cell cocktail of sequential cell infusions targeting CD19 and CD22 may help overcome antigen escape relapse in B-cell malignancies, we therefore conducted an open-label, single-center, and single-arm pilot study of a treatment comprising the sequential infusion of CD22 and CD19 CAR-T cells following autologous hematopoietic stem cell transplantation (HSCT). We aimed to evaluate the safety and efficacy of the treatment in R/R B cell lymphoma patients with residual or progression disease after salvage treatment. Between November 14, 2016 and August 15, 2019, 42 patients underwent BEAM conditioned HDT-ASCT and followed by CAR 19 /22 T cells infusion. The overall response rate was 90.5% (95% CI, 77.4-97.3). At a median follow-up of 13.8 months, the median progression-free survival (PFS) and overall survival (OS) were not reached. The 12-month PFS rate was 85.7 % (95% CI, 70.9 to 93.3). Grade 3 cytokine release syndrome (CRS) occurred in only 2 (5%) patients. 21% patients experienced any grade of neurotoxicity, 5% was severe Grade 3. All CRS and neurotoxicity were reversible. Beyond its extremely high complete response rates, the durability of remission and low toxicity of the autologous HSCT plus CD19/CD22 CAR-T cell cocktail therapy examined in this study emphasize strong potential of such myeloablative conditioning and dual-targeting approaches for R/R B-cell lymphomas. Disclosures No relevant conflicts of interest to declare.

Substituted (2-benzamidothiazol-5-yl)pyrazole-capped AWD*I-NH2 were synthesized and their antimigration activity was studied. The improved efficiency and scalability of the analog synthesis was achieved via a late-stage diversification of the benzoyl group and a convergent route in which the bisazole capping agents and off-resin peptide AWD*I-NH2 were prepared in parallel and coupled together in solution at the last step. Bioassay results indicate that all the peptidomimetics can significantly inhibit the migration of breast cancer cells MDA-MB-231 but possess no apparent cytotoxicity. In general, the antimigration potency of the peptidomimetics is correlated to the electron-withdrawing capacity of the substituents on the terminal phenyl ring. The inhibitory effect shows dose-dependent and holds also against lung and cervical cancer cells. The level of f-actin was reduced dramatically in cells treated with the inhibitor, suggesting that the migration inhibitory effect is related to the disruption of cell locomotive protrusions.

Introduction: Due to the ability to mimic in vivo cellular microenvironments, the development of multicell culture systems has received increasing interest for use as research models and serving as platforms for drug evaluation. Methods: In this study, we developed a perfused microfluidic system to resemble the in vivo intercellular environment and applied it to study the differentiation from neural stem cells into neurons. Results: As determined by immunofluorescence chemistry and quantitative real-time PCR, the neural stem cells grown in this microfluidic system showed an elevated differentiation rate toward the formation of neurons as determined by the increased level of βIII-tubulin production, which is 4 times higher than that of culturing neural stem cells only. Conclusion: These results revealed that some factors secreted into the intercellular microenvironment by adult neuron cells can stimulate the differentiation of neural stem cells, pointing to the importance of developing multicellular culture systems such as the perfused microfluidic system we report here to better resemble the in vivo situation.

L. casei HZ1 was identified from Chinese traditional fermented milk, and angiotensin converting enzyme inhibitory peptide was separated from its culture in our previous work. Here, LGH2 was a novel AMP, identified from the genome of L. casei HZ1. Altogether, roughly 52.76% of LGH2 was &alpha, helical, with the remainder in &beta, strand and random coil in 50% TFE solution tested by CD. The peptide was also an amphipathic and cationic molecule, which was composed of 20 amino acid residues. The similarity of the amino acid sequence between LGH2 and Temporin-RN3 was highest. Then, the peptide successfully expressed in E. coli Rossetta (DE3) pLysS using the SUMO fusion expression system and purified by chromatography technologies. The molecular weight of the peptide was 2448 Da determined by MALDI-TOF MS. Antimicrobial tests showed that the peptide has strong activities against G+ bacteria, special for S. aureus (MIC = 4 &mu, M). The toxicity assay showed that the peptide exhibits a low hemolytic activity against sheep red blood cells. The antimicrobial mechanisms of LGH2 against pathogens were further investigated by dye leakage, CLSM, SEM, and FCM assays. We found that LGH2 can bind to the cell membrane, and destroy its integrity. These significant results indicate that LGH2 has great potential to treat the infections caused by pathogenic bacteria such as S. aureus, and it provides a new template to improve antimicrobial peptides targeting antibiotic-resistant pathogenic bacteria.

Background Hypericum attenuatum Choisy, a traditional Chinese herb, has been shown to be effective in the treatment of diseases associated with inflammation and has been used to treat rheumatic arthritis in China for centuries. However, the underlying mechanism of its anti-inflammatory effect is poorly understood. Hypothesis/Purpose In this study, we aimed to investigate the anti-inflammatory mechanisms of EtOAc fractions of H. attenuatum Choisy (Ha-EtOAc) on lipopolysaccharide (LPS)-induced RAW264.7 macrophage inflammation and hypothesized that Ha-EtOAc could attenuate inflammation in the colon. Study design LPS was utilized to induce RAW264.7 cells inflammation. The anti-inflammatory effect of Ha-EtOAc in RAW264.7 cells was evaluated by measuring the inhibition ratio of nitric oxide (NO) production. Murine ulcerative colitis (UC) was induced by treatment with 2.5% dextran sulfate sodium (DSS). The basic indexes of the mice, including body weight, food intake and hematochezia, were recorded during mice experiments. Methods The expression levels of pro-inflammatory cytokines, including TNF-α, IL-6 and IL-1β, were measured by quantitative real-time PCR and western blot. Additionally, the influences of Ha-EtOAc on the NF-κB and MAPK signaling pathways were determined by western blot and immunofluorescence assays. In addition, the impact of Ha-EtOAc on gut microbiota of mice with UC was detected by 16S rDNA sequencing. Results Ha-EtOAc inhibited the LPS-induced production of NO and decreased the release of TNF-α, IL-6 and IL-1β in RAW264.7 cells in a dose-dependent manner. In addition, pretreatment with Ha-EtOAc could suppress the nuclear translocation of p65 and the phosphorylation of Erk1/2, p38 and JNK. Ha-EtOAc treatment ameliorated murine UC, as reflected by a reduced body weight loss, improved colon shortening, alleviated mucosal damage and decreased releases of pro-inflammatory cytokines. Furthermore, Ha-EtOAc could modulate the composition of microbial communities. Conclusion Our results demonstrated that Ha-EtOAc exhibited anti-inflammatory effects mainly by suppressing the NF-κB and MAPK pathways, and Ha-EtOAc treatment may be a potent therapy for the treatment of ulcerative colitis.

Introduction: Three-dimensional (3D) collagen scaffold models, due to their ability to mimic the tissue and organ structure in vivo, have received increasing interest in drug discovery and toxicity evaluation. Methods: In this study, we developed a perfused 3D model and studied cellular response to cytotoxic drugs in comparison with traditional 2D cell cultures as evaluated by cancer drug cisplatin. Results: Cancer cells grown in perfused 3D environments showed increased levels of reactive oxygen species (ROS) production compared to the 2D culture. As determined by growth analysis, cells in the 3D culture, after forming a spheroid, were more resistant to the cancer drug cisplatin compared to that of the 2D cell culture. In addition, 3D culturing cells showed elevated level of ROS, indicating a physiological change or the formation of a microenvironment that resembles tumor cells in vivo. Conclusions: These data revealed that cellular response to drugs for cells growing in 3D environments are dramatically different from that of 2D cultured cells. Thus, the perfused 3D collagen scaffold model we report here might be a potentially very useful tool for drug analysis.

As a strong transactivator of promoters containing CarG boxes, myocardin‑related transcription factor A (MRTF‑A) is critical for the process of metastasis in tumor cells. Nuclear factor erythroid 2‑like 1 (Nrf1) is well known as an important regulator of oxidative stress, which exists in multiple splicing forms with many unknown functions. The present study demonstrated a novel regulation loop between Nrf1α (the longest splicing form of Nrf1) and MRTF‑A that regulated the migration and invasion of breast cancer MDA‑MB‑231 cells. The underlying mechanism of this regulation look was further investigated. In particular, Nrf1α inhibited migration and invasion of breast cancer cells through inhibiting the expression of MRTF‑A via miR‑219. The current results revealed that miR‑219 could bind to the MRTF‑A 3'‑UTR to directly regulate its expression. However, MRTF‑A could reverse activate the Nrf1α expression through binding to the CarG box in the Nrf1α promoter. It can be speculated that this regulation loop may be a homeostasis mechanism in cells against tumorigenesis.

Multispecies microbial community in natural solid-state fermentation (SSF) is crucial for the formation of Chinese Pu-erh tea's unique quality. However, the association between microbiota and tea quality are still poorly understood. Herein, shotgun metagenomic and metabolomic analysis showed that significant variations in composition of microbiota, collective functional genes, and flavour compounds occurred during SSF process. Furthermore, the formation pathways of the dominant flavours including theabrownin, methoxy-phenolic compound, alcohol and carvone were proposed. Moreover, biological interaction networks analysis among functional core microbiota, functional genes, and dominant flavours indicated Aspergillus was the main flavour-producing microorganism in the early SSF, while many other genera including Bacillus, Rasamsonia, Lichtheimia, Debaryomyces were determined as the functional core microorganism for flavours production in the late SSF. This study provides a perspective for bridging the gap between the microbiota and quality in Pu-erh tea, and benefited for further optimizing production efficiency and product quality.

The cAMP responsive element binding protein 1 (CREB1) is a ubiquitous transcription factor that contributes to the regulation of gluconeogenesis. The mechanisms of the CREB1 function remain largely unknown. In this study, we aimed to explore genetic variations in CREB1 promoter region and determine whether these loci affect transcriptional activity and risk on type 2 diabetes (T2D). Three polymorphisms were identified and designated as MU1, MU2 and MU3, respectively. Genotypic distribution analysis revealed that MU1 genotypes presented similar distribution between T2D and healthy controls (P > 0.05), while the MU2 and MU3 showed significant differences (P

Hypericum ascyron L. (Great St. Johnswort), which belongs to the Hypericaceae family, has been used for the treatment of hematemesis, metrorrhagia, rheumatism, swelling, stomach ache, abscesses, dysentery and irregular menstruation for >2,000 years in China. The aim of the present study was to clarify the anticancer activity compounds from H. ascyron L. and the underlying molecular mechanism. Anticancer activity of H. ascyron L. extract was evaluated using an MTT assay. To confirm the anticancer mechanism of activity compounds, Hoechst 33258, Annexin V-fluorescein isothiocyanate/propidium iodide, 2',7'-dichlorodihydrofluorescein diacetate, rhodamine 123 staining and caspase-3 activity analysis were performed. The results demonstrated that the anti-proliferative action of the mixture of kaempferol 3-O-β-(2″-acetyl) galactopyranoside (K) and quercetin (Q) (molar ratio, 1:1) was significantly increased compared with either of these two compounds separately, and the active fraction of the H. ascyron L. extract |(HALE). HALE, indicating that the anti-proliferative function of H. ascyron L. may be a synergic effect of K and Q. Furthermore, the inhibitory effect of KQ on the growth of HeLa cells was mediated by the induction of apoptosis. To the best of our knowledge, the present study is the first to identify that KQ exhibits significant anti-proliferation activity on HeLa cells via the apoptotic pathway, and is also the first to evaluate the anticancer potential of H. ascyron L. The results of the present study may provide a rational base for the use of H. ascyron L. in the clinic, and shed light on the development of novel anticancer drugs.

As a potential cure for diabetes, more and more attentions have been paid to organ transplants to replace insulin therapy. As a result, many researchers have explored out many programs to get insulin-producing cells (IPCs) to replace the defective β cells. Currently, more and more new induction methods are being proposed, and at the same time, more and more possible induction molecular mechanisms are being revealed. The purpose of this study was to explore whether and how the two factors pdx-1 and myocardin affected the differentiation of rat mesenchymal stem cells (rMSCs) into IPCs. In this study, we investigated the process of transfecting myocardin and/or pdx-1 in rMSCs in vitro. The results showed that rMSCs were able to secrete insulin after cotransfected with myocardin and pdx-1. At the same time, we explored the possible mechanism that myocardin and pdx-1 coinduced rMSCs into IPCs by forming a complex to promote the transcriptional activity of insulin. Our results may provide a theoretical basis to the study of islet transplantation in the future.

The list of primers sequences. (DOCX 29 kb)

Antibodies and antagonists targeting tumour necrosis factor alpha (TNFα) have become a number of the best-selling anti-inflammatory drugs. The nanobody is a single domain antibody derived from the variable domain of a heavy-chain antibody. However, so far, there are still no reports on the recombinant expression of TNFα nanobodies. Here, TNF30, the key anti-TNFα nanobody domain of ozoralizumab, was cloned, expressed in Escherichia coli, and then purified with the Ni2+-chelating affinity chromatography. The western blot results showed that the recombinant TNF30 has strong binding capability with TNFα. Furtermore, the pharmacological and histopathological analysis in the carrageenan-induced mice paw oedema demonstrated that the recombinant nanobodies had potential anti-inflammatory activity in vivo. These works provided a novel approach for the production of TNF30 in E. coli, and might establish a foundation for the industrial production of the drug and the development of more active derivatives against TNFα in the future.

Skin cancer is a major cause of morbidity and mortality worldwide. Mounting evidence shows that exposure of the skin to solar UV radiation results in inflammation, oxidative stress, DNA damage, dysregulation of cellular signaling pathways and immunosuppression thereby resulting in skin cancer. Signal transducer and activator of transcription 3 (STAT3) is well known to function as an anti-apoptotic factor, especially in numerous malignancies, but the relationship between STAT3 activation and DNA damage response in skin cancer is still not fully understood. We now report that STAT3 inhibited DNA damage induced by UV and STAT3 mediated upregulation of GADD45γ and MDC-1 and the phosphorylation of H2AX in UV induced DNA damage. Notably, STAT3 can increase the expression of ATR in A431 cells. Luciferase assay shows that STAT3 activates the transcription of ATR promoter. More importantly, microRNA-383 suppressed ATR expression by targeting 3' (untranslated regions)UTR of ATR in A431 cells, and STAT3 down-regulates the transcription of miR-383 promoter. Thus, these results reveal the new insight that ATR is down-regulated by STAT3-regulated microRNA-383 in A431 cells. Moreover, overexpression of STAT3 enhanced expression of antiapoptosis genes BCL-1 and MCL-1, and depletion of STAT3 sensitized A431 cells to apoptotic cell death following UV. Collectively, these studies suggest that STAT3 may be a potential target for both the prevention and treatment of human skin cancer.

The transcriptional coactivator p300 is highly expressed in breast cancer tissues. MRTF-A is a transcription factor governed by the Rho-GTPase-actin signaling pathway. The purpose of this study was to explore the role of p300 in breast cancer metastasis. Here we showed that the motility of breast cancer cells was enhanced by the overexpression of p300, meanwhile, the transcription of migration-related genes was upregulated. Depletion of p300 downregulated the migration-related genes and slowed down the migration of breast cancer cells. p300 worked synergistically with MRTF-A to activate the transcription of MYH9, MYL9 and CYR61. As identified by co-IP, p300 interacted with the C-terminal TAD domain of MRTF-A. And together with MRTF-A, p300 was associated with the target gene promoters. Furthermore, MRTF-A was found to be acetylated in MCF-7 breast cancer cells. These results demonstrated the involvement of p300 in the MRTF-A mediated gene regulation and breast cancer cell migration.

According to previous studies, hyperoside possesses myocardial protective effects. To investigate whether isoquercitrin and isohyperoside have similar functions, the protective effects of isoquercitrin and isohyperoside against H2O2‑induced injury in H9c2 rat cardiomyoblasts were evaluated using a 3‑(4,5‑dimethylthiazol‑2‑yl)‑2,5‑diphenyltetrazolium bromide assay. The mechanism of action was investigated by assessing the leakage of lactate dehydrogenase (LDH) of hyperoside and isoquercitrin‑pretreated H9c2 cardiomyocytes following H2O2‑induced injury, and examining their effects on the levels of malondialdehyde (MDA) and superoxide dismutase (SOD) activity. The isolation of two flavonoid glycosides from H. ascyron was performed, following extraction, using semi‑preparative high performance liquid chromatography. Using the spectral characteristics, the structures of these compounds were identified as isoquercitrin and isohyperoside. This was the first time, to the best of our knowledge, that isohyperoside has been identified from H. ascyron. The results revealed that isoquercitrin and isohyperoside possessed similar protective effects to hyperoside against H2O2‑induced injury in H9c2 cells. The half maximal inhibitory concentration values of hyperoside, isoquercitrin and isohyperoside were 0.0008, 0.0017 and 0.0002 µM, respectively. Based on these results, isohyperoside possessed more marked protective effects against H2O2‑induced injury in the H9c2 cardiomyoblasts. The significant reduction in LDH leakage, decrease in MDA level and increase in SOD activity also provided evidence of the cardioprotective effects of isoquercitrin and isohyperoside. The present study reported for the first time, to the best of our knowledge, the myocardial protective effects of isoquercitrin and isohyperoside. The mechanism of action may involve protection of the cell membrane from oxidative damage.

Hypericum ascyron L. has been used as a traditional medicine for the treatment of wounds, swelling, headache, nausea and abscesses in China for thousands of years. However, modern pharmacological studies are still necessary to provide a scientific basis to substantiate their traditional use. In this study, the mechanism underlying the antimicrobial effect of the antibacterial activity compounds from H. ascyron L. was investigated. Bioguided fractionation of the extract from H. ascyron L. afforded antibacterial activity fraction 8. The results of cup plate analysis and MTT assay showed that the MIC and MBC of fraction 8 is 5 mg/mL. Furthermore, using Annexin V-FITC/PI, TUNEL labeling and DNA gel electrophoresis, we found that cell death with apoptosis features similar to those in eucaryon could be induced in bacteria strains after exposure to the antibacterial activity compounds from H. ascyron L. at moderate concentration. In addition, we further found fraction 8 could disrupt the cell membrane potential indicate that fraction 8 exerts pro-apoptotic effects through a membrane-mediated apoptosis pathway. Finally, quercetin and kaempferol 3-O-β-(2″-acetyl)-galactopyranoside, were identified from fraction 8 by means of Mass spectrometry and Nuclear magnetic resonance. To our best knowledge, this study is the first to show that Kaempferol 3-O-β-(2″-acetyl)-galactopyranoside coupled with quercetin had significant antibacterial activity via apoptosis pathway, and it is also the first report that Kaempferol 3-O-β-(2″-acetyl)-galactopyranoside was found in clusiacea. Our data might provide a rational base for the use of H. ascyron L. in clinical, and throw light on the development of novel antibacterial drugs.

Ethnopharmacological relevance Hypericum ascyron L. ( H. ascyron L.) has been used as a traditional medicine for the treatment of wounds, swelling, headache, nausea, stomach ache, abscesses, dysentery and chronic bronchitis. Pharmacological studies are necessary to provide a scientific basis to substantiate their traditional use. In this study, antimicrobial effect and its mechanism of the H. ascyron L. extract was investigated. Materials and methods Antimicrobial activity of the H. ascyron L. extract was evaluated by the 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay (MTT). To confirm the antimicrobial mechanism of H. ascyron L. extract on bacterial cells, Annexin V/propidium iodide (PI) double staining, TUNEL, and DNA laddering analysis were performed. Results Antibacterial effects of the H. ascyron L. extract were tested on Enterobacter cloacae , Klebsiella pneumonia , Escherichia coli , Staphylococcus aureus and Micrococcus luteus , and the results showed that the sensitive bacteria of H. ascyron L. extract mainly included E. coli, S. aureus and M. luteus , especially M. luteus . The MBC value of H. ascyron L. extract on M. luteus was equal to the MIC (20 mg/mL). H. ascyron L. extract could induce cells death of M. luteus through apoptosis pathway. Conclusions This study provides scientific support for some of the traditional uses and the pharmacological activities of H. ascyron L. Our data provide a rational base for the folkloric use of H. ascyron L.

Breast cancer is the leading cause of cancer death in women worldwide which is closely related to metastasis. But the exact molecular mechanism of metastasis is still not fully understood. We now report that both MRTF-A and STAT3 play important roles in migration of MDA-MB-231 breast cancer cells. Moreover, MRTF-A and STAT3 synergistically increased MDA-MB-231 cell migration by promoting the expression of migration markers urokinase-type plasminogen activator (uPA) and osteopontin (OPN) and inhibiting the expression of breast cancer metastasis suppressor 1 (BRMS1). Luciferase reporter assays demonstrated that MRTF-A and STAT3 do not affect transcription of the BRMS1 promoter. Instead, we identified a newly molecular mechanism by which MRTF-A and STAT3 synergistically controlled MDA-MB-231 cell migration by recruiting DNMT1 to hypermethylate the promoter of BRMS1 and thus affect the expression of BRMS1. Interestingly, physical interaction between MRTF-A and STAT3 synergistically promotes the transactivity of DNMT1 by binding to the GAS element within the DNMT1 promoter. Our data thus provide important and novel insights into the roles of MRTF-A and STAT3 in regulating MDA-MB-231 cell migration. © 2015 IUBMB Life, 67(3):202–217, 2015

Separation and purification of hypocrellin A (HA) and hypocrellin B (HB) are conducted for extracted concentrated solution of hypocrellin through dry packing of molecularly imprinted polymers (MIPs) of hypocrellin A and B and preparative solid phase extraction column. The result shows that the optimum packing column combination for mixed adsorption is H5 (i.e. MIP of hypocrellin A and B are respectively 10.5g and 4.5g), and that the elution sequence is hypocrellin A elution first and then hypocrellin B elution next after the eluent is changed.

Hypertrophic growth of cardiomyocytes in response to pressure overload is an important stage during the development of many cardiac diseases. Ca(2+) overload as well as subsequent activation of Ca(2+) signaling pathways has been reported to induce cardiac hypertrophy. Myocardin, a transcription cofactor of serum response factor (SRF), is a key transducer of hypertrophic signals. However, the direct role of myocardin in Ca(2+) signal-induced cardiomyocyte hypertrophy has not been explained clearly. In the present study, we discovered that embryonic rat heart-derived H9c2 cells responded to the stimulation of calcium ionophore A23187 with a cell surface area enlargement and an increased expression of cardiac hypertrophy marker genes. Increased Ca(2+) also induces an organization of sarcomeres in neonatal rat cardiomyocytes, as revealed by α-actinin staining. Increased Ca(2+) could upregulate the expression of myocardin. Knockdown of myocardin by shRNA attenuates hypertrophic responses triggered by increased intracellular Ca(2+), suggesting that Ca(2+) signals induce cardiomyocyte hypertrophy partly through activation of myocardin. Furthermore, A23187 treatment directly activates myocardin promoter, chelation of Ca(2+) by EGTA inhibits this activation and knockdown of myocardin expression using shRNA also abrogates A23187-induced ANF and SK-α-actin promoter activity. CSA (calcineurin inhibitor) and KN93 (CaMKII inhibitor) inhibit A23187-induced the increase in myocardin expression. These results suggest that myocardin plays a critical role in Ca(2+) signal-induced cardiomyocyte hypertrophy, which may serve as a novel mechanism that is important for cardiac hypertrophy.

In this paper, a heart-cutting two-dimensional high-performance liquid chromatography coupled with the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method was established for controlling the quality of different batches of Hypericum ascyron extract for the first time. In comparison with the common one-dimensional fingerprint, the second-dimensional fingerprint compiled additional spectral data and was hence more informative. The quality of H. ascyron extract was further evaluated by similarity measures and the same results were achieved, the correlation coefficients of the similarity of ten batches of H. ascyron extract were ＞0.99. Furthermore, we also evaluated the quality of the ten batches of H. ascyron extract by antibacterial activity. The result demonstrated that the quality of the ten batches of H. ascyron extract was not significantly different by MTT. Finally, we demonstrated that the second-dimensional fingerprint coupled with the MTT method was a more powerful tool to characterize the quality of samples of batch to batch. Therefore the proposed method could be used to comprehensively conduct the quality control of traditional Chinese medicines.

With the rapid development of pharmaceutical industry and the requirement of economic globalization, the demand of international pharmaceutical professionals in China has been kept increasing. In this project, to set up a Chinese-English bilingual teaching system of pharmacology for undergraduate students majored in Pharmaceutical Engineering, a survey had been conducting to evaluate the general talents and attitudes of students and the need of the industry for 7 years in Tianjin University of Science and Technology, the teaching contents were then adjusted and optimized to highlight the application ability of professional English on the basis of the emphasis of pharmacology knowledge. Basing on these works, a novel Chinese-English bilingual textbook of pharmacology was published in 2013, and an open and multi-mode reformed Chinese-English bilingual education system was also established in the university. In our opinion, this novel textbook and the teaching system might throw light to the bilingual education of pharmacology in China.

Background Long non-coding RNAs (lncRNAs) have emerged as a new and crucial layer of gene regulation in recent years and regulate various biological processes such as carcinogenesis and metastasis. LncRNA HOTAIR, an oncogenic lncRNA, is involved in human tumorigenesis and dysregulated in cervical cancer. Megakaryoblastic leukemia 1 (MKL1), as a transcription coactivity factor, involved in cancer metastasis and cell differentiation. However, the precise mechanism of biological roles of HOTAIR and MKL1 in cancer cells remain unclear. Methods The expression levels of HOTAIR and MKL1 were measured by quantitative PCR (qPCR), immunoblotting, in situ hybridization (ISH) and immunohistochemistry (IHC). Wound-healing and transwell assays were used to examine the invasive abilities of HeLa cells. Luciferase reporter assays and CHIP were used to determine how MKL1 regulates HOTAIR. Tissue microarray and immunohistochemical staining were used to assess the correlation between HOTAIR and MKL1 in Cervical cancer tissues in vivo. Result In this study, we have identified that MKL1 had a role in the induction of migration and invasion in cervical cancer cells. Moreover, the expression level of MKL1, as the targeting gene of miR206, was decreased after HOTAIR inhibition in HeLa cells. Agreement with it, Highly level of MKL1 correlation with HOTAIR is validated in cervical cancer tissues. Importantly, HOTAIR is observed to participate in the silencing of miR206 expression. Interestingly, HOTAIR inhibition could also accelerate the expression of MKL1 in cytoplasm. What is more, MKL1 can activate the transcription of HOTAIR through binding the CArG box in the promoter of HOTAIR. Conclusion These elucidates that the phenotypic effects of migration and invasion observed after HOTAIR inhibition, at least in part, through the regulation of MKL1 via inhibition of miR206 expression in HeLa cells. These data indicate the existence of a positive feedback loop between HOTAIR and MKL1. Together, these findings suggest that MKL1 is an important player in the functions of HOTAIR in the migration and invasion of cancer cells. Electronic supplementary material The online version of this article (10.1186/s12964-018-0216-3) contains supplementary material, which is available to authorized users.

As a strong transactivator of promoters containing CarG boxes, myocardin was critical for the cardiac muscle program and necessary for normal cardiogenesis. So it probably represents a viable therapeutic biomarker in the setting of cardiac hypertrophy and failure. In recent years, the studies of regulation of cardiac hypertrophy via myocardin are so common, and the molecular mechanism is becoming more and more clear. Here, we have revealed a kind of interaction between mhrt and myocardin shown as a feedback regulatory mechanism in the regulation of cardiac hypertrophy. That is, the lncRNA mhrt can affect the acetylation of myocardin by HDAC5 to inhibit cardiac hypertrophy induced by myocardin. Moreover, myocardin also can directly activate the mhrt transcription through binding to the CarG box. Thus, mhrt and myocardin form a regulation loop in the process of cardiac hypertrophy. This finding may play a positive role in revealing the complete mechanisms of cardiac hypertrophy.

Bacillus sp. N16-5 is an alkaliphile with a great ability to utilize mannan. Its mannan utilization gene cluster has been identified in a previous study. The ManR protein encoded by the cluster was predicted to be a LacI family regulator, and the transcription level of the mannan utilization gene cluster was upregulated after the manR gene was deleted, indicating that ManR is the repressor of this cluster. The transcription of the related genes was downregulated when manH, encoding the extracellular substrate-binding domain of the manno-oligosaccharide transporter, was deleted. Furthermore, isothermal titration calorimetry revealed that mannotetraose and mannopentose are ligands of ManR. These results all corroborate the hypothesis that the mannan utilization gene cluster is repressed by the transcription regulator ManR, and that the repression is removed when it binds to manno-oligosaccharides, which are generated by mannan degradation and transported into the cell by a specific transporter.

The re-expression of a fetal cardiac gene program, including bran natriuretic peptide (BNP), contributes to the development of cardiac hypertrophy. The mRNA and protein expression of BNP are increased in hypertrophic and failing hearts. Myocardin, a co-activator of serum response factor (SRF), potently transactivates hypertrophic genes via binding the CArG box within the promoter of these hypertrophic genes and induces cardiomyocyte hypertrophy. The neuronrestrictive silencer factor (NRSF), also known as RE-1 silencing transcription factor (REST), has been shown to negatively regulate the gene transcription of BNP by binding to a neuron-restrictive silencer element (NRSE). In the present study, the gene fragment of human BNP gene promoter containing two CArG boxes and one NRSE was cloned into empty vector pGL3-Basic to construct BNP promoter luciferase reporter plasmid. Myocardin expression plasmids or NRSF expression plasmids were co-transfected with BNP promoter-luc plasmids into COS-7 cells to analyze the regulatory effects of myocardin and NRSF on the promoter activity of BNP. The results showed that myocardin over-expression significantly up-regulated the activity of BNP promoter, while NRSF inhibited myocardin-induced activation of BNP promoter. The results suggest that NRSF may play a critical role in regulating BNP gene in cardiac hypertrophy and heart failure.

Maca and probiotics are benefit to people. In order to combine the functions of maca and probiotics, a maca-probiotics compound granule was prepared in our previous study. However, whether the probiotics could survive well in vivo has been remained unclear. To address this problem, the present study analyzed the acid resistance, salt tolerance and heat tolerance of the five probiotics strains in the compound granule. The results showed that the number of viable bacteria would be decreased when the maca-probiotics compound granule was treated with low pH (pH ≤ 3.5) for 4 h or 0.05–0.3% bile salts for 2 h, but there still have a certain survivors could resist these rigorous conditions. The number of survivors began to decrease after 2 h at pH 3.0. Besides, when the product was treated with different temperature (25–52 °C) for 10 min, the reduction in the number of viable bacteria would be accelerated significantly when the temperature exceed 47 °C. These results suggested that the probiotics strains in the maca-probiotics compound granule still have acceptable resistance ability to acid, bile salts and heat, indicating that they could exhibite expected functions after oral administration. In addition, maca-probiotics compound granule should be stored at low temperature and be taken after meals with water below 47 °C.

Idiopathic thrombocytopenic purpura (ITP) is a chronic acquired organ-specific autoimmune hemorrhagic disease, It has been proved that the reduce of regulatory T cells (Treg) (CD4+CD25+FOXP3+) was an important cause of ITP, and the transcriptional factor FOXP3 played an important role in the proliferation and differentiation of Treg cells. However, the definite transcription mechanism of foxp3 gene is unclear. In this study, human foxp3 promoter luciferase reporter construct was successfully constructed. Then transfected one of the two overexpression vectors of human MKL-1 and human STAT5A or both into 293T cells, and detected the activation of the foxp3 promoter by luciferase reporter assay. The result showed that transfected each of the overexpression vectors can enhance the transcriptional activity of FOXP3, and this activity effect can be enhancing when the two vectors were cotransfected. Our research will further reveal the proliferation and differentiation of Treg cells and may provide theoretical foundations for methods to cure ITP.

Breast cancer is the main cause of cancer deaths in women around the world, and is closely related to metastasis. We know that MRTF-A and STAT3 play an important role in the migration of breast cancer. Vimentin is an important marker gene for epithelial-mesenchymal transition (EMT), which is closely related to the process of breast cancer metastasis. The construction of the mutant luciferase reporter gene plasmid will demonstrate that MRTF-A and STAT3 can modulate the transcription of vimentin. In this study, the Vimentin promoter luciferase reporter gene was derived from PCR amplification of the Vimentin promoter. The PCR amplified fragment was cloned into the PGL-3 vector by restriction enzyme digestion. The promoter sequence was confirmed by sequencing. The sequencing results showed that the Vimentin promoter luciferase reporter plasmid was constructed successfully. MDA-MB-231 cells were respectively transfected with the Vimentin luciferase reporter plasmids and pcDNA3.1 and pcDNA3.1-MRTF-A and pcDNA3.1-STAT3, and were analyzed by luciferase reporter assay. The results showed that MRTF-A could significantly enhance the transcriptional activity of vimentin gene promoter compared with transfected pcDNA3.1 group, while STAT3 group could slightly up-regulate the transcriptional activity of vimentin gene promoter.

Tip60 is a cellular protein belonging to the MYST family of histone acetyl transferase (HAT). It functions in DNA damage repair and transcriptional regulation in eukaryotic cells. Human papillomavirus (HPV) is a causative factor of cervical cancer which threatens the health of females worldwide. In the present study, the encoding sequence of Tip60 was amplified with the RT-PCR method and cloned into expressing vector pcDNA-tag2B to generate a Tip60-expressing plasmid. Subsequently, the effect of Tip60 on HPV18 E6 E7 genes was investigated in HeLa cells which carrying the integrated HPV18 DNA fragments. Results of plasmid sequencing confirmed the correct cloning of Tip60 plasmid and then the efficient overexpression of Tip60 in HeLa cells was demonstrated with RT-qPCR and Western-blotting. With Tip60 overexpression, the expression of HPV18 E6 and E7 genes was repressed significantly. Furthermore, co-overexpression of Tip60 and EP400, a chromatin remodeling factor, in HeLa cells synergistically suppressed the expression of HPV18 E6-E7. Findings in this study would help to reveal the roles of Tip60 in the regulation of viral oncogenes and inspire targeted therapy of cervical cancer.

To explore the function of probiotic bacteria in puer tea pile fermentation which was implemented by a symbiosis of many fermenting microbes and enzymes, the species, abundances and dynamics of probiotic bacteria were quantitatively investigated. Using the real-time quantitative PCR (qPCR) primers specifically targeting different genera bacteria, the results showed that Enterococcus spp., Lactococcus spp., Lactobacillus group and Bacillus spp. were all found at 0, 15, 30, and 45 days during the fermentation, but Bifidobacterium spp. was not involved throughout the fermentation. Furthermore, among these four genera, Lactococcus spp. was the predominant probiotic with population of 1.68 × 109–2.78 × 109 copy g−1 in the fermentation, followed by Enterococcus spp. harboring 3.43 × 104–6.25 × 105 copy g−1, and Lactobacillus group and Bacillus spp. had 4.26 × 103–5.17 × 104 and 2.21 × 103–2.31 × 105 copy g−1, respectively. In addition, the dynamics of four genera probiotic bacteria and universal bacteria were explored to show completely different and complex change trends with no significant correlation with pH of tea and each other. Population dynamic studies of probiotic bacteria revealed their ecological feature and importance for puer tea pile fermentation, and also facilitated further exploration of probiotic resources and health benefits of puer tea.

Xylanase, as critical hemicellulose degrading enzyme, it could degrade xylan into oligosaccharides and has been widely used in medicine, paper-making, animal forage, marine functional foods and textile industry. So exploring new xylanase with excellent enzymatic characteristic will be benefit for improving production efficiency and reducing cost. In this study, a novel xylanase of family 10 (Xyn13-3) was obtained from the genomic DNA of alkaline soil by touchdown-PCR and thermal asymmetric interlaced (TAIL) PCR methods, which shares the highest identity (61%) with the reported GH10 xylanase (XP_018659952.1) in GenBank database. Xyn13-3 contained a catalytic motif of GH10 (339 residues). The recombinant plasmid pPIC9-Xyn13-3 was constructed and the recombinant protein was successfully expressed in Pichia pastoris GS115. The special activity of recombinant xylanase was 135.24 U/mg. Exploring the use of novel enzymes from different organisms, with distinct characteristics, could contribute to the development of a highly efficient system for lignocellulose conversion.

Cervical cancer which is identified as the second most common type of cancer in women worldwide, is responsible for more than 275,100 mortalities each year and associated with the long-term infection of high-risk human papillomavirus, including HPV18. HPV18 DNA contains two vital oncogenes, E6 and E7, which have crucial roles in malignant transformation in cervical cancer. Previous studies have revealed that LncRNAs (long non-coding RNAs), which are non-coding transcripts that are more than 200nt in length, are important regulators in various biological processes and abnormalities in LncRNA expression are correlated with progression of cervical cancer. In this article, we examined 9 cancer-related LncRNAs and 2 newly identified LncRNAs in cancer or non-cancer cell lines. Further, we enforced the expression of HPV18 E6 and E7 in HeLa cervical cancer cells or introduced HPV18 E6 and E7 encoding plasmids into HaCaT non-cancer cells to explore the relationship between HPV18 E6, E7 and the expression of cancer-related LncRNAs. The results showed that the expression level of these LncRNAs varied in different cell lines, and the HPV18 E6 and E7 regulated the expression levels of some cancer related LncRNAs, in particular MALAT1 and TMOPO2.

In this study, we collected 6 dairy samples from Xinyuan County, Yili Kazakh Autonomous Prefecture, Xinjiang Uygur Autonomous Region, China. These samples are all traditional Kazak dairy product including ghee, dry yogurt, butter, yogurt milk cheese and red knots. Total 22 strains were isolated and identified by 16SrDNA sequencing and their phylogenetic tree was constructed according to the 16SrDNA sequences. These strains belong to 5 genera, including Lactobacillus, Enterococcus, Leuconostoc, Acetobacter, and Bacillus. Among them Lactobacillus and Enterococcus contain most abundant strains, and L. plantarum and A. orientalis were the most widespread strains. L. plantarum A1-3-15 was found to exhibit good bile salt and acid tolerance. This study indicated that the traditional Kazak dairy products exhibit good biodiversity and are good resources of probiotics.

Purpose We previously found that the histone methyltransferase suppressor of variegation, enhancer of zeste, trithorax and myeloid-nervy-deformed epidermal autoregulatory factor-1 domain-containing protein 3 (SMYD3) is a potential independent predictive factor or prognostic factor for overall survival in gastric cancer patients, but its roles seem to differ from those in other cancers. Therefore, in this study, the detailed functions of SMYD3 in cell proliferation and migration in gastric cancer were examined. Materials and Methods SMYD3 was overexpressed or suppressed by transfection with an expression plasmid or siRNA, and a wound healing migration assay and Transwell assay were performed to detect the migration and invasion ability of gastric cancer cells. Additionally, an MTT assay and clonogenic assay were performed to evaluate cell proliferation, and a cell cycle analysis was performed by propidium iodide staining. Furthermore, the expression of genes implicated in the ataxia telangiectasia mutated (ATM) pathway and proteins involved in cell cycle regulation were detected by polymerase chain reaction and western blot analyses. Results Compared with control cells, gastric cancer cells transfected with si-SMYD3 showed lower migration and invasion abilities (P

MRTF-A is a transcriptional co-activator being critical for multiple processes including tissue fibrosis and cancer metastasis. The Rho-actin signaling stimulates the nuclear translocation and transcriptional activity of MRTF-A with little effect on the expression of MRTF-A gene. High expression of MRTF-A was observed in pancreatic cancer tissues and in TGF-β treated breast cancer cells. However, the mechanism for the upregulation of MRTF-A gene remains unclear. In this study, we showed that the transcription of MRTF-A was regulated by the Wnt-β-catenin signaling in breast cancer cells. LiCl treatment, Wnt3a treatment or β-catenin overexpression enhanced the transcription of MRTF-A gene. In agreement, depletion of β-catenin with siRNA diminished MRTF-A transcription. With ChIP assays, β-catenin was identified to interact with the MRTF-A promoter whereby it increased histone H4 acetylation and RNA polymerase II association. Further, results of RT-qPCR and Western-blotting supported that the transcriptional co-activator activity of MRTF-A was controlled by both the Rho-actin and the Wnt-β-catenin signaling pathways. MRTF-A was required for cell migration stimulated by the Wnt-β-catenin signaling. Taken together, our results suggest that MRTF-A integrates the Rho-actin and the Wnt-β-catenin signaling to regulate migration-related genes and consequently increases the mobility of breast cancer cells.

Breast cancer is the leading cause of cancer death in women worldwide which is closely related to metastasis. Our previous study has shown that MRTF-A promote the migration of MDA-MB-231 cells and WDR1 promotes breast cancer cell migration. But the exact molecular mechanism on metastasis is still not fully understood, we now report that WDR1 enhanced the effect of MRTF-A induced-MDA-MB-231 cell migration by promoting the expression of the EMT markers and migration markers via RhoA-MRTF-A signaling pathway. Importantly, WDR1 promoted the nuclear importion of MRTF-A by affecting the expression of nuclear transport protein importin. But WDR1 did not affect the expression of MRTF-A. Interestingly, MRTF-A promoted the expression of miR-206 via its promoter CArG box but miR-206 inhibits the migration of breast cancer cells through suppressing the expression of WDR1 and MRTF-A via targeted their 3'UTR. Our data thus provide important and novel insights into MRTF-A-miR-206-WDR1 form feedback loop to regulate breast cancer cell migration.

High risk HPV infection is a causative factor of cervical cancer. The constitutive expression of HPV E6-E7 genes is important for the maintenance of cancer phenotypes. The cellular transcription co-activator p300 plays a crucial role in the regulation of HPV genes thus it was targeted for the inhibition of HPV-associated cervical cancer. In the present study, HPV positive cervical cells were treated with C646, a selective inhibitor of p300, to investigate its influence on HPV E6-E7 expression and cancer cell growth. Results of RT-qPCR, Western-blot and promoter activity assays showed that C646 inhibited the transcription of HPV E6-E7, which was accompanied with the accumulation of p53 protein. Meanwhile, cell proliferation was suppressed, glucose metabolism was disrupted and apoptosis was induced via the intrinsic pathway. Generally, the anti-cervical cancer potential of C646 was demonstrated and a novel mechanism was proposed in this study.

We report the complete genome sequence of Lactobacillus plantarum CGMCC 8198, a novel probiotic strain isolated from fermented herbage. We have determined the complete genome sequence of strain L. plantarum CGMCC 8198, which consists of genes that are likely to be involved in dairy fermentation and that have probiotic qualities.

We report the complete genome sequence of Lactobacillus plantarum CGMCC 8198, a novel probiotic strain isolated from fermented herbage. We have determined the complete genome sequence of strain L. plantarum CGMCC 8198, which consists of genes that are likely to be involved in dairy fermentation and that have probiotic qualities.

Myocardin is well known to play a key role in the development of cardiomyocyte hypertrophy. But the exact molecular mechanism regulating myocardin stability and transactivity to affect cardiomyocyte hypertrophy has not been studied clearly. We now report that NF-κB (p65) can inhibit myocardin-induced cardiomyocyte hypertrophy. Then we explore the molecular mechanism of this response. First, we show that p65 can functionally repress myocardin transcriptional activity and also reduce the protein expression of myocardin. Second, the function of myocardin can be regulated by epigenetic modifications. Myocardin sumoylation is known to transactivate cardiac genes, but whether p65 can inhibit SUMO modification of myocardin is still not clear. Our data show that p65 weakens myocardin transcriptional activity through attenuating SUMO modification of myocardin by SUMO1/PIAS1, thereby impairing myocardin-mediated cardiomyocyte hypertrophy. Furthermore, the expression of myocardin can be regulated by several microRNAs, which play important roles in the development and function of the heart and muscle. We next investigated potential role of miR-1 in cardiac hypotrophy. Our results show that p65 can upregulate the level of miR-1 and miR-1 can decrease protein expression of myocardin in cardiac myocytes. Notably, miR-1 expression is also controlled by myocardin, leading to a feedback loop. These data thus provide important and novel insights into the function that p65 inhibits myocardin-mediated cardiomyocyte hypertrophy by downregulating the expression and SUMO modification of myocardin and enhancing the expression of miR-1.

Breast cancer is the leading cause of cancer death in women worldwide which is closely related to metastasis. But the exact molecular mechanism on metastasis is still not fully understood; we now report that both MRTF-A and STAT3 play important role in breast cancer migration of MDA-MB-231 cells. Moreover, MRTF-A and STAT3 synergistically increased MDA-MB-231 cell migration by promoting the expression of migration markers Myl-9 and Cyr-61. Importantly, we identified a detailed molecular mechanism of MDA-MB-231 cell migration controlled via physical interaction between MRTF-A and STAT3, which synergistically promote the transactivity of the migration marker Myl-9 and Cyr-61 by CArG box binding. Interestingly, the two signaling pathways RhoA-MRTF-A and JAK-STAT3 across talk to regulate MDA-MB-231 cell migration. Our data thus provide important and novel insights into the roles of MRTF-A and STAT3 in regulating MDA-MB-231 cell migration.

Human cytomegalovirus (HCMV) may increase the incidence of restenosis and predispose to atherosclerosis. The lesions of restenosis and atherosclerosis often contain smooth muscle cells (SMCs) with high rates of proliferation and apoptosis. One of the immediate early (IE) gene products of HCMV-IE2 affects transcriptional activities of some cellular factors in SMCs, including myocardin. In this study, we studied the effects of IE2 and myocardin on PI3K pathway inducer wortmannin induced apoptosis in rat aortic SMCs. We show that the transcriptional activity of myocardin on Mcl-1 promoter is enhanced by co-expression of HCMV IE2 in rat aortic SMCs; and the expressions of mRNA and protein of antiapoptotic genes-Mcl-1 and Bcl-2 are upregulated by IE2 alone and co-transfection of myocardin and IE2, but decreased by myocardin-specific shRNA in rat aortic SMCs. We further demonstrate that co-expression of myocardin and HCMV IE2 declines apoptotic cell numbers and caspase-3 activities induced by serum starvation plus wortmannin in rat aortic SMCs. The results suggest that HCMV IE2 enhances myocardin-mediated survival of rat aortic SMCs under serum deprivation and PI3-kinase inhibition, partly via activation of Mcl-1's antiapoptosis effect. Our study connects HCMV IE2 to myocardin-induced transcriptional program for rat aortic SMCs survival and proliferation, involving in HCMV related restenosis and atherosclerosis.