Objective To study the success rate of ventilator weaning in acute exacerbation of chronic obstructive pulmonary disease (AECOPD) patients influenced by bobbed endotracheal intubation. Methods One hundred and sixteen AECOPD patients who were given invasive mechanical ventilation and reached the standards of off-ventilator were divided into control group (general endotracheal intubation group) and experiment group bobbed (tracheal intubation) by random digits table method with 58 cases each. The respiratory rate (RR), heart rate (HR), rapid and shallow breathing index (RSBI), oxygenation index (OI), pulse blood oxygen saturation (SpO2), tidal volume and success rate of ventilator weaning were compared. Both groups were implemented of extubation if they reached the standards of the extubation time by observing their ability in spontaneous breathing test (SBT). Results The RR, HR and RSBI before extubation in experiment group were significantly lower than those in control group:(19.7 ± 2.3) times/min vs. (23.5 ± 2.3) times/min, (91.2 ± 6.3) times/min vs. (93.4 ± 8.1) times/min and 80.2 ± 6.7 vs. 90.5 ± 9.6, P<0.05, and the OI, SpO2, tidal volume and success rate of ventilator weaning were significantly higher than those in control group: (269 ± 9) mmHg (1 mmHg = 0.133 kPa) vs. (245 ± 16) mmHg, 0.929 ± 0.014 vs. 0.870 ± 0.037, (6.1 ± 1.2) ml/kg vs. (5.1 ± 0.8) ml/kg and 91.38%(53/58) vs. 77.59%(45/58), P<0.05. Conclusions The bobbed endotracheal intubations can improve the success rate of ventilator weaning in patients with AECOPD.

BACKGROUND:Studies have confirmed that the nano-hydroxyapatite/chitosan/alginate composite materials have a certain flexibility and strength and possess a bioactivity similar to human bone.
OBJECTIVE:To explore the effect of the nano-hydroxyapatite/chitosan/alginate composite materials on the repair of rabbit mandible defects.
METHODS:Bilateral mandibular defect models of 10 mm × 5 mm × 5 mm were made in 18 healthy New Zealand white rabbits. Then, the rabbits were divided into two groups:experimental group was implanted with nano-hydroxyapatite/chitosan/alginate composite material, and control group was implanted with hydroxyapatite/chitosan composite. At 4, 8 and 12 weeks after implantation, cone-beam CT was applied to observe implant degradation, cal us growth and bone connection in the defect area;new bone formation was observed by hematoxylin-eosin staining.
RESULTS AND CONCLUSION:The gray values of the bone density in the experimental group and control group gradual y increased with time, and there were remarkably significant differences between the two groups at different time points (P<0.01). At the same time point, the experimental group was superior to the control group in gross observation, cone-beam CT observation, gray value of CT and histological observation (P<0.05). At 4-8 weeks after implantation, the implant materials in the two groups were gradual y degraded with a blurred junction between the defect and bone tissue, and a smal amount of new bone formed tightly integrated to the recipient bone tissue, in which the experimental group was more significant. And during 8 to 12 weeks, the degradation of implanted materials in the two groups was basical y complete, and the implant began to merge with the recipient bone tissue, with further generation of new bone tissue and gradual repair of bone defect area, in which, the experimental group was more obvious. Results show that the nano-hydroxyapatite/chitosan/alginate can repair bone defects effectively, and promote the new bone formation.

Objective To investigate the neuroprotective effects of neurotrophin-3 (NT-3) expression controlled by five copies of the hypoxia-responsive elements after focal cerebral ischemia .Methods Three groups of rats re-ceived RV-5H-NT3, RV-5H-EGFP or saline injection .Three days after gene transfer , the rats underwent 90 min of transient middle cerebral artery occlusion ( tMCAO) , followed by 1-28 days of reperfusion .Immunohistostaining and western blotting were performed to detect ischemia/hypoxia-regulated expression of NT-3 controlled by HRE . The volume of brain infarction and the apoptosis were analysised by TTC and TUNEL staining .The neurological scoring was determined by neurological behavior tests .Results Three days after tMCAO , brain NT-3 expression was significantly increased in the RV-5HNT3-transduced animals compared with the RV-5H-EGFP or saline group (P<0.05), and brain infarct volume was smaller in the RV-5H-NT3-transduced group than the RV-5H-EGFP or saline group ( P<0.05 ) .The percentage of TUNEL-positive cells was reduced in RV-5 H-NT3-transduced brains compared with the RV-5 HEGFP or saline group 3 and 7 days after tMCAO ( P<0.05 ) .Furthermore , the neurolog-ical status of RV-5H-NT3-transduced rats was better than that of RV-5H-EGFP-or saline-transduced animals from 1 day to 4 weeks after tMCAO ( P<0.05 ) .Conclusions HRE may modulate NT-3 expression in the ischemic brain tissue and that the up-regulated NT-3 may effectively improve neurological status following tMCAO due to de-creased initial damage .

Objective To construct and identify the recombinant retroviral vector containing five copies of hypoxia responsive elements (5HRE)and neurotrophin-3 (NT-3 ).Methods Using PCR,enzyme digestion and DNA ligase,5HRE and human derived NT-3 were cloned into the retroviral vector plasmid (pLNCX)to construct the recombinant retroviral vector plasmid pLNCX-5HRE-SV40-NT3-IRES-EGFP.The retrovirus RV-5HRE-NT3 was packaged in the PT67 cells,and then it was purified and concentrated by high-speed centrifugation.After infected for 48 h with the concentrated retrovirus,the number of the EGFP positive cells in the NIH 3T3 cells was counted by fluorescence activated cells and sorted to calculate the retrovirus titer.Results The retroviral vector plasmid,pLNCX-5HRE-SV40-NT3-IRES-EGFP,was successfully constructed,and the retrovirus was packaged and defined as RV-5HRE-NT3.After purification and concentration,the retrovirus titer reached 9.1 × 10 6 cfu/mL. Conclusion The recombinant retroviral vector which carried out hypoxia-regulated expression of NT-3 was successfully constructed.It may provide basis for studies on hypoxia-regulated expression of the exogenous genes.

Objective To explore the effect of Ligustrazine on neurogenesis in cortex after focal cerebral ischemia in rats. Methods Focal cerebral ischemia was induced by left middle cerebral arteryocclusion with asuture. Two hours later, injection of Ligustrazine (80 mg/kg, 1 time/d) was performed peritoneally. Four hours after the ischemia,5-bromodeoxyuridine (BrdU) (50 mg/kg, 1 time/d) was injected peritoneally. At 7 d, 14 d and 21 d after ischemia,BrdU positive cells in the cortex were observed by immunohistochemical staining. Results In ischemic model group, at 7 day, sparsely-distributed BrdU positive cells were observed in the Ⅱ - Ⅵ layers of the ipsilateral cortex, with a band-like distribution in ischemic penumbra. With the prolongation of ischemia, the number of BrdU positive cells increased.In Ligustrazine group, BrdU positive cells were also observed in the Ⅱ - Ⅵ layers of the cortex, with an intense distribution in ischemic penumbra. The numbers of BrdU positive cells at 7 d, 14 d and 21 d were more than those in ischemic model group respectively. Conclusion Ligustrazine increases the proliferated cells in cortex after focal cerebral ischemia in rats. The results suggest that it may be useful for promoting self-repair after ischemia.

In most mammalian central nervous system diseases, axons are damaged.Due to the limited ability of damaged neurons to promote axonal regeneration, the formation of glial scar and the release of inhibitory nutrients, it is difficult to regenerate axons of damaged neurons. The purpose of this study was to investigate the effect of cerebroprotein hydrolysate for injection (II) (CBL) on neuritogenesis and its underlying mechanism. Immunofluorescence staining was used to detect the axon length of mouse neuroma cells (Neuro-2a) and mouse primary cortical neuronal cells. Western blotting was used to detect the expression of phosphorylated TrkB protein in Neuro-2a cells and mouse primary cortical neuronal cells. The results showed that CBL could increase the axon length of Neuro-2a cells or mouse primary cortical neuronal cells, and that the phosphorylation level of TrkB in neuronal cells was significantly increased when 5 μg/mL CBL was applied to neuronal cells for 1 h. In conclusion, CBL can promote neuritogenesis, and increase the expression of phosphorylated TrkB, which may be related to the activation of TrkB signaling pathway.

OBJECTIVETo explore the effects of salidroside (sal) on the expressions of Bcl-2, Bax and caspases-3 proteins in cultured rat subventricular zone (SVZ) neural stem cells (NSCs) exposed to hypoxia injury.

METHODSPrimarily cultured SVZ NSCs from adult SD rats were incubated with salidroside (120 and 240 µmol/L) for 24 h prior to exposure to hypoxia. The cell viability was assessed with MTT assay, and the cell apoptosis was analyzed using TUNEL staining and flow cytometry. Western blotting was performed to detect the expressions of Bcl-2, Bax and caspase-3 in the cells.

RESULTSSalidroside pretreatment of the cells for 24 h resulted in an obvious resistance to hypoxia-induced cell apoptosis and decrement of cell viability (P<0.05). Salidroside also antagonized the effect of hypoxia exposure in lowering Bcl-2/Bax ratio apoptosis of rat neural stem cells and decreased the expression of caspases-3 protein (P<0.05).

CONCLUSIONSalidroside can significantly resist hypoxia-induced. The neuroprotective effect of salidroside may be related to the modulation of expressions of apoptosis-related proteins.

Animals ; Caspase 3 ; metabolism ; Cell Hypoxia ; Cells, Cultured ; Flow Cytometry ; Glucosides ; pharmacology ; Neural Stem Cells ; drug effects ; Phenols ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats ; Rats, Sprague-Dawley ; bcl-2-Associated X Protein ; metabolism

The cDNA of cattle Ghrelin gene was amplified from abomasum fundic gland mRNA of Qinchuan Cattle by RT-PCR. PCR product was cloned into the T vector pGM-T to construct pGh-T1 for sequencing. Then the cDNA was subcloned into the prokaryotic expressing plasmid vector pET32a (+) and transformed into host Escherichia coli strain BL21 (DE3) for expression. The expression of pGh-32 mature Ghrelin protein was induced by IPTG and was identified by SDS-PAGE. The expression product was observed with soluble protein and inclusion body. Western blotting showed that the recombinant protein was recognized by his-antibody specifically. The protein was purified by Ni-NTA column and was used to inject rabbits to obtain polyclona antibody. ELISA result showed that the antibody titer was 1:12 800. The immunohistochemistry test between the hypothalamus arcuate nucleus and the antibody showed that fusion protein had biological activity. This will provide a basis for further study on the biological function of Ghrelin protein to growth and development and fat deposition of cattle.
Animals ; Cattle ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Ghrelin ; genetics ; metabolism ; Recombinant Fusion Proteins ; genetics ; metabolism

Blood-brain barrier (BBB) damage after ischemia significantly influences stroke outcome. Compound LFHP-1c was previously discovered with neuroprotective role in stroke model, but its mechanism of action on protection of BBB disruption after stroke remains unknown. Here, we show that LFHP-1c, as a direct PGAM5 inhibitor, prevented BBB disruption after transient middle cerebral artery occlusion (tMCAO) in rats. Mechanistically, LFHP-1c binding with endothelial PGAM5 not only inhibited the PGAM5 phosphatase activity, but also reduced the interaction of PGAM5 with NRF2, which facilitated nuclear translocation of NRF2 to prevent BBB disruption from ischemia. Furthermore, LFHP-1c administration by targeting PGAM5 shows a trend toward reduced infarct volume, brain edema and neurological deficits in nonhuman primate