Improvement of Left Ventricular Remodelling by Inhibition of NF-κB in a Rat Model of Myocardial Infarction.

BACKGROUND: To investigate the effects of inhibition of NF-κB activation on left ventricular (LV) remodelling in a rat model of myocardial infarction (MI). METHODS: The acute MI model was established by ligation of left anterior descending coronary artery. Pyrrolidine dithiocarbamate (PDTC) (20mg/kg, Qd) was administered intraperitoneally to inhibit NF-κB activation. Eight weeks later, the cardiac structure and LV ejection fraction were assessed with echocardiography. The rat body, heart, and LV weights were measured to calculate LV mass indices. Activation of NF-κB in non-infarcted myocardium was detected by a TransAM NF-κB p65 Transcription Factor Assay Kit. Cardiac collagen volume fraction was evaluated by Masson staining. RESULTS: Eight weeks after the MI model was established, the LV posterior wall thickness in PDTC and MI group was 1.75±0.07mm and 1.85±0.07mm respectively (p<0.05). The LV mass index in the PDTC group (2.53±0.09) was lower than in the MI group (2.65±0.08, p<0.05). The LVEF in the PDTC group (63.89%±4.21%) was higher than in the MI group (42.73%±8.94%, p<0.05). The interstitial collagen deposition in the non-infarcted myocardium in the PDTC group was less than in the MI group (7.25%±1.88% vs. 10.09%±2.19%, p<0.05). CONCLUSION: Inhibition of activation of NF-κB may result in improvement of myocardial remodelling after myocardial infarction, which is possibly attributable to reduced collagen deposition in non-infarcted areas.

Septicemic melioidosis: a case report and literature review.

This report was about a 22-year-old male patient who died from acute and severe septicemic melioidosis. He was initially misdiagnosed as having pneumonia in a local hospital in Hainan but transferred to Guangzhou Red Cross Hospital after ineffective treatment and detection of Burkholderia pseudomallei in his blood sample. Unfortunately, right diagnosis and antibiotic treatment of melioidosis had been delayed so that his condition deteriorated rapidly out of control. The victim's death highlights the importance of early diagnosis and effective antibiotic treatment of septicemic melioidosis in areas where this disease is prevalent.

The inhibitory effect of salvianolic acid B on TGF-ß1-induced proliferation and differentiation in lung fibroblasts.

Salvianolic acid B (Sal B), one of the major water-soluble compounds of Danshen (a popular Chinese herb), possesses many of the biological activities, such as antifibrogenic effect in liver and renal diseases. Transforming growth factor-ß1 (TGF-ß1) plays a central role in the development of pulmonary fibrosis by stimulating extracellular matrix (ECM) accumulation and activating fibroblasts. Here, we investigated the effects of Sal B on cell proliferation, collagen synthesis, endogenous TGF-ß1 production, and α-smooth muscle actin (α-SMA, a marker of myofibroblasts) expression in human lung fibroblasts stimulated by TGF-ß1 in vitro. The cell proliferation rates were analyzed by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide) assay. The expression of TGF-ß1 and type I collagen at both the mRNA and protein levels was detected by reverse transcription polymerase chain reaction (RT-PCR), enzyme-linked immunosorbent assay (ELISA), and radioimmunoassay, respectively. The α-SMA expression was detected by Western blot. TGF-ß1 treatment of lung fibroblasts increased cell proliferation rates, and enhanced the expression level of type I collagen, endogenous TGF-ß1 production, and α-SMA expression (P < .05). The treatment with only Sal B did not affect the proliferation and differentiation of lung fibroblasts. Interestingly, Sal B was found to inhibit TGF-ß1-induced cell proliferation, expression of type I collagen, endogenous TGF-ß1 production, and α-SMA expression in lung fibroblasts. Moreover, the inhibitory effect of Sal B on TGF-ß1-induced proliferation and differentiation in lung fibroblasts was more significant when treated with high-dose Sal B (1 µmol/L versus 10 µmol/L, P < .05). These data demonstrate that Sal B inhibits TGF-ß1-induced cell proliferation and differentiation in vitro experiment.

[Effects of Notch signaling pathway and vascular endothelial growth factor gene on the functions of endothelial cells derived from rat bone marrow mesenchymal stem cells].

OBJECTIVE: To explore the effects of Notch signaling pathway and the vascular endothelial growth factor [VEGF(165)] gene on the functions of endothelial cells derived from rat bone marrow mesenchymal stem cells (MSCs). METHODS: Isolated and cultivated rat bone marrow MSCs in vitro, then the cells were treated by VEGF165 and basic fibroblast growth factor (bFGF) for 2 weeks to induce them to differentiate into endothelial cells. The gene of VEGF165 was transfected into differentiated endothelial cells to promote the functions of the cells. The receptor Notch1 and the ligand Jagged1 of the Notch signaling were detected by reverse transcription-polymerase chain reaction (RT-PCR) before and after the transfection. γ-secretase inhibitor L-685458 was used to block Notch pathway. Migration ability of cells was detected by scarification test. Cells were inoculated on semisolid gel to study their ability of forming capillary-like structure. RESULTS: After transfection, VEGF165 mRNA could be detected on the differentiated endothelial cells. The expression of Jagged1 mRNA was up regulated(1.08 ± 0.01 vs. 1.01 ± 0.02,P < 0.01) and there was no change in Notch1 mRNA(0.60 ± 0.02 vs. 0.59 ± 0.01,P > 0.05). The ability of migration was enhanced (number of cells on the scratched area:46.45 ± 4.46 vs. 41.61 ± 1.42,P < 0.05), and the ability of forming capillary-like structure on semisolid gel showed no change (cells classification: 3.00 ± 0.89 vs. 2.00 ± 0.89,P > 0.05). After the transfection, using the γ-secretase inhibitor L-685458 to block the Notch signaling transduction, the ability of migration of the differentiated endothelial cells (number of cells on the scratched area: 51.72 ± 3.47 vs. 46.45 ± 4.46,P < 0.05), and that of forming capillary--like structure (cells classification: 4.17 ± 0.75 vs. 3.00 ± 0.89, P < 0.05), was also further enhanced. CONCLUSION: Transfection of the gene of VEGF165 into the differentiated endothelial cells can reinforce the function of these cells, and when Notch signaling was blocked, this effect can be further amplified.

[The role of Notch signaling during the differentiation of rat bone marrow mesenchymal stem cells into endothelial cells].

OBJECTIVE: To research the role of Notch signaling during the differentiation of bone marrow mesenchymal stem cells (MSCs) into endothelial cells and its effect on the functions of the differentiated cells. METHODS: Rat bone marrow MSCs were isolated and cultured in vitro, then the cells were treated with vascular endothelial growth factor (VEGF165) and basic fibroblast growth factor (bFGF) for 2 weeks to induce it to differentiate into endothelial cells. The differentiated cells were identified by fluorescence immunoassay. The receptors and ligands of the Notch signaling were detected by reverse transcription-polymerase chain reaction (RT-PCR) before and after the differentiation. γ-secretase inhibitor was used to block Notch pathway. Migration ability of cells were assessed by scarification test. Cells were inoculated on semisolid gel to study their ability of forming the capillary-like structure. RESULTS: After inducing MSCs to differentiate into endothelial cells by VEGF165 and bFGF, MSCs gained the characteristics of the endothelial cells with expression of CD31 and Flk1. There were Notch1 mRNA and Jagged1 mRNA expressions in rat bone marrow MSCs. The expression changes in the receptor Notch1 were not statistically significant on the differentiated cells (0.59±0.01 vs. 0.59±0.01, P>0.05), but there was a trend towards an increase of Jagged1 mRNA (1.01±0.02 vs. 0.99±0.03, P>0.05). When Notch pathway was blocked, the differentiated cells' migration ability was increased (number of cells on the scratched area: 44.61±4.34 vs. 40.06±2.43, P<0.05), and the ability of forming capillary-like structure was also increased (cells classification: 3.67±0.82 vs. 2.00±0.89, P<0.01). CONCLUSION: Notch signaling may have an important role during the differentiation of MSCs into endothelial cells. The function of differentiated cells were strengthened when Notch pathway was blocked.

[The effects of the plasma levels of serotonin and neuropeptide on ventilated patients with Yitongshu acupuncture points injection].

OBJECTIVE: To evaluate the change in the plasma levels of serotonin (5-hydroxytryptamine) and neuropeptide (beta-endorphin, beta-EP) after injection of Yitongshu into Zusanli points in patients under mechanical ventilation. METHODS: Twenty-eight patients undergoing mechanical ventilation were randomly divided into two groups: midazolam combined with fentanyl group (control group, n=14) and midazolam combined with fentanyl and Yitongshu group (Yitongshu group, n=14). The drugs were given to the patients continuously intravenously with an injection pump in an even rate, with the dosage adjusted to reach the sedative target of visual analog score (VAS)< or =3-4 and Ramsay 2-4. Yitongshu injection (4 ml) was injected into the Zusanli point on both sides 11 hours or 23 hours after administration of the durgs in Yitongshu group. The hemodynamic and respiratory parameters, including mean arterial pressure (MAP), heart rate (HR), respiratory rate (RR), pulse oxygen saturation (SpO(2)), oxygenation index (PaO(2)/FiO(2)) and pressure airway (Paw), were recorded, and the sedation levels (VAS and Ramsay) were evaluated before sedation and 1, 12, 24 hours after sedation in these patients. The plasma levels of 5-hydroxytryptamine and beta-EP were examined before sedation, 12 hours and 24 hours after sedation. RESULTS: Compared with that before sedation, HR and VAS score were significantly lower, and Ramsay score was significantly higher in both groups. MAP was significantly lower at 1 hour, and RR at 12 hours and 24 hours , as well as the Paw at 24 hours, and the PaO(2)/FiO(2) was significantly higher at 24 hours. The level of 5-hydroxytryptamine at 12 hours and 24 hours in Yitongshu group [(101.45+/-14.67) ng/L, (104.86+/-11.74) ng/L] was significantly higher than that in control group [(61.57+/-10.62) ng/L, (59.86+/-8.64) ng/L, both P<0.05]. But the level of beta-EP showed no difference between two groups [control group: (162.72+/-38.44) ng/L at 12 hours, (151.83+/-24.54) ng/L at 24 hours; Yitongshu group: (169.35+/-28.10) ng/L at 12 hours, (159.41+/-15.89) ng/L at 24 hours, both P>0.05]. CONCLUSION: Yitongshu injection can reduce the plasma level of 5-hydroxytryptamine in ventilated patients, but with no effect on beta-EP.

Modulatory effect of CCK-8S on GABA-induced depolarization from rat dorsal root ganglion.

CCK is a brain-gut peptide that is abundantly distributed in both gastrointestinal tract and mammalian brain. The sulfated octapeptide fragment of cholecystokinin (CCK-8S) has been shown to be involved in numerous physiological functions such as behavior, anxiety, learning/memory processes and neuropathic pain. CCK-8S is one of the strongest endogenous anti-opioid substances and suppresses opioid peptides-mediated 'pre-synaptic inhibition' of gamma-aminobutyric acid (GABA) release. Here we provide evidence that CCK-8S modulates GABA-evoked membrane depolarization in rat dorsal root ganglion (DRG) neurons using intracellular recording technique. Bath application CCK-8S-induced membrane depolarization in most of the rat DRG neurons. The depolarization was blocked by prolumide but not LY225910. Pretreatment with CCK-8S suppressed the GABA-evoked depolarization in a concentration-dependent manner. The CCK-8S inhibition was also time-dependent and reached the peak at about 2 min. The inhibitory effect of CCK-8S was strongly suppressed by pre-incubation of CCK-B receptor antagonist LY225910, phospholipase C inhibitor U73122, protein kinase C inhibitor chelerythrine and calcium chelator BAPTA-AM, respectively. The protein kinase A inhibitor H-89 did not affect CCK-8S effect. The results suggest that CCK-8S inhibits GABA-A receptor function by activation of CCK-B receptor followed by activation of intracellular PLC-Ca(2+)-PKC cascade. Thus, CCK-8S might enhance nociceptive information transmission through inhibition of the "pre-synaptic inhibition" evoked by GABA, which may explain its role in modulation of primary sensory information (especially pain).