SOX6 AU controls myogenesis by cis-modulation of SOX6 in cattle.

Long non-coding RNAs (lncRNAs) have been shown to be involved in the regulation of skeletal muscle development through multiple mechanisms. The present study revealed that the lncRNA SOX6 AU (SRY-box transcription factor 6 antisense upstream) is reverse transcribed from upstream of the bovine sex-determining region Y (SRY)-related high-mobility-group box 6 (SOX6) gene. SOX6 AU was significantly differentially expressed in muscle tissue among different developmental stages in Xianan cattle. Subsequently, knockdown and overexpression experiments discovered that SOX6 AU promoted primary skeletal muscle cells proliferation, apoptosis, and differentiation in bovine. The overexpression of SOX6 AU in bovine primary skeletal muscle cells resulted in 483 differentially expressed genes (DEGs), including 224 upregulated DEGs and 259 downregulated DEGs. GO functional annotation analysis showed that muscle development-related biological processes such as muscle structure development and muscle cell proliferation were significantly enriched. KEGG pathway analysis revealed that the PI3K/AKT and MAPK signaling pathways were important pathways for DEG enrichment. Notably, we found that SOX6 AU inhibited the mRNA and protein expression levels of the SOX6 gene. Moreover, knockdown of the SOX6 gene promoted the proliferation and apoptosis of bovine primary skeletal muscle cells. Finally, we showed that SOX6 AU promoted the proliferation and apoptosis of bovine primary skeletal muscle cells by cis-modulation of SOX6 in cattle. This work illustrates our discovery of the molecular mechanisms underlying the regulation of SOX6 AU in the development of beef.

[Research progress on the role of FOXOs family in cancer].

The Forkhead box class O proteins (FOXOs) family consists of highly conserved transcription factors, including FOXO1, FOXO3, FOXO4 and FOXO6. Each member of the FOXOs family is ubiquitously expressed and involved in regulating many biological activities such as tumor cell proliferation, apoptosis, migration and oxidative stress. The activity of FOXOs is mainly regulated by post-translational modification, and its inactivation is mainly mediated by the over-activation of its upstream modifying enzymes, which provides a possibility to use drugs to recover its activity. It is worth noting that FOXOs can not only inhibit, but also promote the occurrence and development of human tumors due to the complex effects of FOXOs. This review will summarize the structure and activity regulation of FOXOs, and discuss their tumor inhibiting effects by limiting cell proliferation and inducing apoptosis, as well as their tumor promoting effects by maintaining cell homeostasis, promoting metastasis and inducing drug resistance, so as to provide new ideas for the pathological research of related diseases and open up new ways to promote broader prevention and treatment strategies.

Population pharmacokinetic analysis for dabigatran etexilate in Chinese patients with non-valvular atrial fibrillation.

We aimed to develop a pharmacokinetic (PK) and pharmacodynamic (PD) model from healthy Chinese subjects and real-world non-valvular atrial fibrillation (NVAF) patients. We also investigated meaningful intrinsic and extrinsic factors and related biomarkers for bleeding events. We characterized the integrated PK/PD models based on rich PK/PD data [dabigatran concentration, activated partial thromboplastin time (APTT), prothrombin time (PT), and anti-factor IIa (anti-FIIa) activity] from 118 healthy volunteers and sparse PD data [APTT, PT, and anti-FIIa] from 167 patients with NVAF after verifying the model extrapolation performance. We also documented the correlations between PD biomarkers and clinically relevant bleeding events over one year. Next, we used the final integrated PK/PD model (a two-compartment, linear model with first-order absorption) to evaluate the influence of dosage and individual covariates on PD parameters. The age, high-density liptein cholesterol (HDL-C), and creatinine clearance (CrCL) improved the PK model fit. The linear direct-effects PD model described the correlation between APTT, PT, and anti-FIIa and plasma concentration. CrCL improved the PD model fit. Anti-FIIa was more sensitive to the increase in dabigatran exposure than APTT and PT in the PD model. Therefore, fixed dabigatran doses could be prescribed for patients with NVAF without adjusting for age and HDL-C. We observed an elevated bleeding tendency with higher peak and trough values of APTT, PT, and anti-FIIa. Randomized studies should be performed to evaluate the efficacy and safety of low-dose dabigatran in Chinese patients with NVAF.

Intestinal injury following liver transplantation was mediated by TLR4/NF-κB activation-induced cell apoptosis.

Intestinal motility and barriers are often impaired due to intestinal congestion during liver transplantation. Intestinal bacteria and enterogenous endotoxins enter into the blood stream or lymphatic system and translocate to other organs, which can result in postoperative multi-organ dysfunction (MODF) and systemic inflammatory reaction syndrome (SIRS) severely affecting patient survival. However, the mechanisms underlying liver transplantation-induced intestinal injury remain unclear and effective therapies are lacking. Thus, the present study investigated whether these effects were associated with endotoxin-mediated apoptosis. Rat autologous orthotopic liver transplantation (AOLT) models were established to observe dynamic intestinal injuries at different time-points following reperfusion. Changes in the levels of endotoxins and the primary receptor, toll-like receptor 4 (TLR4), as well as its downstream signaling molecule, nuclear factor-κB (NF-κB) were all determined. Finally, immunohistochemistry and terminal deoxynucleotidyl transferase dUTP nick end labeling assays were conducted to detect caspase-3 expression and intestinal cell apoptosis, respectively. AOLT resulted in significant pathological intestinal injury, with the most serious intestine damage apparent four or eight hours following reperfusion. Furthermore, the levels of endotoxins and inflammatory cytokines, such as tumor necrosis factor-α and interleukin-6, peaked during this time period and gradually decreased to the normal level. Notably, TLR4 and downstream NF-κB expression, as well as NF-κB-mediated caspase-3 activation and intestinal cell apoptosis coincided with the intestinal pathological damage. Thus, the possible mechanism of post-liver transplantation intestinal injury was demonstrated to be associated with NF-κB activation-induced cell apoptosis.

[Cisplatin enhances apoptosis in bladder cancer cells via autophagy].

OBJECTIVE: To monitor the cisplatin-induced autophagy and investigate the function of autophagy in bladder cancer cells. METHODS: The transmission electron microscope was used to detect autophagic vacuoles and the fluorescence microscope to detect GFP-LC3. The expressions of proteins, such as LC3, PARP, mTOR, P70S6K were analyzed by immunoblotting. Cell viability was analyzed by MTS assay, in which rapamycin was used to inhibit mTOR phosphorylation and enhance autophagy. LC3 expression was knocked down by RNA interference. RESULTS: In bladder cancer cell T24, autophagic vacuoles were observed by the transmission electron microscope and GFP-LC3 aggregation was viewed by the fluorescence microscope after cisplatin treatment. The LC3-II accumulation was enhanced by cispaltin treatment. Particularly at the concentrations of 50, and 100 µmol/L for 48 h , the gray value of LC3-II/Actin(%) increased 30 and 44, respectively. Cisplatin treatment inhibited the phosphorylation of mTOR/P70S6K, which was most significant at the concentration of 100 µmol/L for 48 h. Cisplatin also induced cell viability loss, which was 12% and 45% at the concentrations of 50, and 100 µmol/L for 24 h. This effect could be enhanced by rapamycin (F=74.890,P<0.01). Furthermore, knocking down LC3 by RNA interference reduced PARP cleavage. CONCLUSION: Cisplatin could induce autophagy in bladder cancer cell T24, which promoted cisplatin-induced apoptosis.

Panax notoginseng saponins enhances the cytotoxicity of cisplatin via increasing gap junction intercellular communication.

Panax Notoginseng Saponins (PNS) have been well known to have anti-tumor activity and enhance cytotoxicity of some cancer chemotherapy agents, but the mechanisms underlying these effects are still unknown. This study investigates the effect of PNS on cytotoxicity of cisplatin and the relationship between this effect and the modulation of gap junctions (GJ) function by PNS in a transfected cell line. The cytotoxicity of cisplatin (0.25-1 µg/mL) was increased in the presence of GJ. Inhibition of gap junction by either GJ blocker or interception of Connexin (Cx) expression decreased the cytotoxicity of cisplatin. Increasing GJ function enhanced cytotoxicity of cisplatin, only in the cells with functional GJ. PNS (50-200 µg/mL) significantly enhanced cisplatin cytotoxicity, but this effect required functional gap junctions between the cells. Exposure of the cells to PNS (50-200 µg/mL) for 4 h leads to a significant enhance in dye coupling of GJ in a dose-dependent manner. These results suggest that PNS increases the cytotoxicity of cisplatin through enhancement of GJ activity.

The effects of 2-aminoethoxydiphenyl borate and diphenylboronic anhydride on gap junctions composed of Connexin43 in TM4 sertoli cells.

2-Aminoethoxydiphenyl borate (2-APB) has recently been demonstrated to inhibit gap junction (GJ) channels, whereas the underlying mechanisms are still unknown. Using mouse TM4 Sertoli cell which expresses connexin43 (Cx43), we explored the effects of 2-APB and its analogues on dye-coupling through junctional channels formed by Cx43 and on expression of Cx43. Exposure of the cells to 2-APB (1-50 µM) and one of its analogues diphenylboronic anhydride (DPBA) (1-30 µM) for 4 h leads to a significant decrease in dye coupling of GJ in a concentration-dependent manner. The inhibitory effects of 2-APB and DPBA are reversible since decreased GJ coupling resumes after the two compounds are washed out. The disfunction of GJ induced by 2-APB and DPBA is associated with a decrease in total amount of Cx43 protein and number of GJs on the cell membrane. 2-APB and DPBA do not alter Cx43 phosphorylation state and the level of Cx43 mRNA expression. The loss of Cx43 protein is prevented by either lysosomal or proteasomal inhibitor, suggesting that the decrease in Cx43 results from a 2-APB or DPBA-enhanced degradation of Cx43. The present results indicate that 2-APB and DPBA inhibit GJ communication through decreasing Cx43 expression in TM4 cells.

Berberine potentizes apoptosis induced by X-rays irradiation probably through modulation of gap junctions.

BACKGROUND: Clinical combination of some traditional Chinese medical herbs, including berberine, with irradiation is demonstrated to improve efficacy of tumor radiotherapy, yet the mechanisms for such effect remain largely unknown. The present study investigated the effect of berberine on apoptosis induced by X-rays irradiation and the relation between this effect and gap junction intercellular communication (GJIC). METHODS: The role of gap junctions in the modulation of X-rays irradiation-induced apoptosis was explored by manipulation of connexin (Cx) expression, and gap junction function, using oleamide, a GJIC inhibitor, and berberine. RESULTS: In transfected HeLa cells, Cx32 expression increased apoptosis induced by X-rays irradiation, while inhibition of gap junction by oleamide reduced the irradiation responses, indicating the dependence of X-rays irradiation-induced apoptosis on GJIC. Berberine, at the concentrations without cytotoxicity, enhanced apoptosis induced by irradiation only in the presence of functional gap junctions. CONCLUSIONS: These results suggest that berberine potentizes cell apoptosis induced by X-rays irradiation, probably through enhancement of gap junction activity.

[Dynamic membrane bioreactor with glass fiber tube covered with organic membrane].

A novel grille form complex membrane module composed of glass fiber covered with organic membrane and the dynamic membrane bioreactor (DMBR) with this complex membrane were studied. The results showed that the flux of the dynamic membrane of glass fiber tube without covering with organic membrane solution was only 4 L/(m2 x h) at a trans-membrane pressure (TMP) of 0.02 MPa. After the modification of covering with the organic membrane solution, the complex dynamic membrane flux could reach to a level of 16 L/(m2 x h) at a TMP of 0.01 MPa in operation, and after a hydraulic and chemical cleaning, the membrane flux was up to 17.1 L/(m2 x h) at a lower TMP of 0.003 MPa. When the glass fiber tube was coated with a membrane solution with a concentration of 1:4 (membrane materials/solution in volume ratio), the flux of the complex membrane worked steadily at 14.29 L/(m2 x h) more than 51 days, and according to calculation by TMP rising, the flux could maintain for almost 275 d. The average removal rates of COD and NH4+ -N were 81.96% and 83.66% respectively by the DMBR, and that were 21.01% and 3.61% only by the complex dynamic membrane. Moreover, the cost of complex membrane was approximately 40-60 yuan/m2, which was lower than the traditional organic membrane.

[Study of a new type of tubular self-forming dynamic membrane bioreactor and its application for treatment of landfill leachate].

A new type of fiberglass tubular self-forming dynamic membrane bioreactor (DMBR) and its application for treatment of landfill leachate on laboratory scale were studied. The results showed that the system worked with a average membrane flux 3.75 L/m2 x h) maintained by gravity filtration at a trans-membrane pressure (TMP) of 2 900 Pa for near 80 days. After the modification of membrane module, a higher membrane flux was achieved at a TMP of 1 450 Pa and the membrane flux could be maintained steadily at 6 L/(m2 x h) for a long time. At the same time, the formation and filtration performance of dynamic membrane (DM) was tested. The effluent turbidity was blow 1.0 NTU and the average removal of COD, BOD5 and NH4(+) -N in this system exceeded 71%, 96% and 98% respectively. Moreover, COD removal from the supernatant on an average of 19.34% was made only by the dynamic membrane.