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Objective:To observe the effect of lipopolysaccharide (LPS) induced conditioned medium of alveolar epithelial cells on the inflammatory response and cell damage of vascular endothelial cells, and explore its mechanism.Methods:The LPS induced type Ⅱ alveolar epithelial cells (A549) conditioned medium was used as a stimulus to induce human umbilical vein endothelial cells (HUVEC) damage. The cell counting kit-8 (CCK-8) was used to detect the effect of 0% (blank group), 12.5%, 25%, 50%, 75% and 100% A549 cell conditioned medium cultured for 6, 12, 24 and 48 hours on the cell viability of HUVEC. Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of inflammatory factors [interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α)] and vasoactive substances [vascular endothelial growth factor (VEGF), endothelin-1 (ET-1)] in the supernatant. Phalloidin staining was used to observe the effects of A549 cells conditioned medium on cell morphology. The expressions of protein kinase B/nuclear factor-κB (AKT/NF-κB) pathway in HUVEC induced by conditioned medium was detected by Western blotting.Results:Compared with the blank group, A549 cells conditioned medium with concentrations of 12.5%, 25%, and 50% had no significant effects on cell viability of HUVEC after 6, 12, and 24 hours, but the activity of HUVEC decreased significantly after 48 hours. Therefore, 12.5%, 25%, 50% A549 cell conditioned medium stimulated for 24 hours was selected as the induction condition for follow-up experiments. Compared with the blank group, the level of IL-6 was significantly increased in 12.5% and 50% conditioned medium groups (ng/L: 2?438.95±64.89, 3?036.41±96.69 vs. 1?736.75±20.99, both P < 0.05), the level of TNF-α was significantly increased in 12.5% and 25% conditioned medium groups (ng/L: 174.08±11.09, 81.37±8.17 vs. 50.03±0.26, both P < 0.01), the levels of VEGF and ET-1 were significantly increased in 12.5%, 25% and 50% conditioned medium groups [VEGF (ng/L): 173.60±41.44, 192.49±12.38, 318.89±27.90 vs. 66.68±19.65; ET-1 (ng/L): 54.88±1.37, 36.69±0.29, 24.07±0.73 vs. 10.67±0.25, all P < 0.01]. Phalloidin staining showed that HUVEC induced by 25% A549 cells conditioned medium were irregular in shape, uneven in size, disordered in arrangement, widened in gap, dense and unclear in microfilament structure and serrated in cell membrane. Furthermore, the average fluorescence intensity of 25% conditioned medium group significantly increased compared to the blank group (67?205.60±3?430.40 vs. 56?272.67±7?650.95, P < 0.05). Western blotting showed that compared with the blank group, the expression of HUVEC cells phosphonated inhibitor α of NF-κB (p-IκBα) was significantly decreased in the 12.5%, 25%, and 50% conditioned medium groups (p-IκBα/IκBα: 0.38±0.08, 0.67±0.12, 0.31±0.07 vs. 1.00±0.00, all P < 0.01), the expressions of phosphonated-AKT (p-AKT) and VEGF were significantly increased (p-AKT/AKT: 1.50±0.18, 1.42±0.27, 1.61±0.14 vs. 1.00±0.00, VEGF/GAPDH: 1.37±0.10, 1.53±0.22, 1.40±0.12 vs. 1.00±0.00, all P < 0.05), the expression of phosphonated NF-κB p65 (p-P65) was significantly increased in the 25% conditioned medium group (p-P65/P65: 1.45±0.14 vs. 1.00±0.00, P < 0.05). Conclusion:LPS induced conditional culture medium of alveolar epithelial cells induced endothelial cell damage via activating AKT/NF-κB pathway.
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N6-methyladenosine (m6A) is one of the most common post-transcriptional modifications of eukaryotic mRNA. The m6A modification accelerates mRNA metabolism and translation, and plays an important role in cell differentiation, embryonic development and stress response. As a reversible epigenetic modification, m6A modification plays an important role in many physiological and pathological processes. The m6A modification is closely related to the occurrence and progression of respiratory diseases, and the m6A modification regulatory factor may be a potential target for regulating respiratory diseases. This article reviews the role of m6A modification in the development of respiratory diseases such as lung cancer, acute lung injury (ALI), asthma, pulmonary fibrosis, and chronic obstructive pulmonary disease (COPD). The purpose of m6A modification is to provide a reference for the pathogenesis of respiratory diseases and the study of targets.
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Objective:To determine the content of triptolide in different parts of Tripterygii radix by high performance liquid chromatography. Methods:Tripterygii radix was determined by high performance liquid chromatography with Agilent Technologies C18 column (4.6 mm × 250 mm, 5 μm), acetonitrile-water (33:67) as mobile phase, flow rate of 1 ml/min, the column temperature of 30 ℃, injection volume of 20 ml and wavelength of 218 nm. Results:The linear relationship of triptolide was good in the range of 0.204-2.040 μg ( r=0.999 9), and the average recovery rate was 93.35%; RSD was 1.56% ( n=6). The lignin content in different parts of root was higher than that of the skin. Conclusions:The method is simple, rapid and accurate, which could be used to determine the content of triptolide in Tripterygii radix.
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OBJECTIVE： To investigate the effects of Kangfuxin liquid on repairing cartilage defect model of knee osteoarthritis （KOA） in rabbits and its mechanism. METHODS： Totally 72 male New Zealand rabbits were selected and randomly divided into model control group and Kangfuxin low-dose， medium-dose， high-dose groups， with 18 rabbits in each group. A cartilage defect model of the medial femoral condyle of the right knee joint in rabbits was established by drilling after anesthesia surgery. Then the rabbits in each group were given medicine via articular cavity immediately. Kangfuxin low-dose， middle-dose and high-dose groups were given 20％， 40％， 80％ Kangfuxin liquid； model control group was given constant volume of normal saline consecutively， 0.2 mL/kg， once every 3 days. At 4th， 8th， 12th week after medication， the wound repair of cartilage defect in rabbits was observed. Immediately after medication and at 4th， 8th， 12th week after medication， repaired tissue of cartilage defect in rabbits was scored histologically with Wakitani scoring standard under light microscope. At 12th week after medication， pathological changes of repaired tissue of cartilage defect in rabbits were observed by Masson staining. The levels of NO， SOD and LPO in joint fluid and PYD in urine of rabbits were detected by ELISA. RESULTS： At 4th， 8th， 12th week after medication， compared with model control group， cartilage defects in rabbits were repaired well in Kangfuxin low-dose， medium-dose and high-dose groups. At 4th， 8th， 12th week after medication， compared with immediately after medication and model control group at same time point， histomorphological score of repairing cartilage defect of knee joint in rabbits decreased significantly in Kangfuxin low-dose， medium-dose and high-dose groups （P＜0.05）. At 12th week after medication， compared with model control group， the histopathology degree of cartilage defect of knee joint in rabbits was significantly alleviated in Kangfuxin low-dose， medium-dose and high-dose groups. At 4th， 8th， 12th week after medication， compared with model control group， the levels of NO and LPO in joint fluid and PYD level in urine were decreased to different extent in Kangfuxin low-dose， medium-dose and high-dose groups， while SOD level was increased to different extent； at 12th week after medication， the difference of each index has statistical significance （P＜0.05 or P＜0.01）. CONCLUSIONS： Kangangxin liquid can significantly repair cartilage defect of KOA cartilage defect model rabbits， the mechanism of which may be associated with increasing the expression of SOD and mediating NO-inhibited chondrocyte apoptosis.
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Objective To elevate the efficacy and safety of descending neurogenic evoked potentials (DNEP) monitoring during severe rigid spinal deformity surgery.Methods All of 108 patients (43 males,65 females) who underwent surgical treatment for spinal deformity in our spinal center from July 2010 to August 2013 were retrospectively reviewed.The average age (17.5±5.8) ys(range 12-50 ys),the average following period is 38.6 months(range 24-52 months).Combined monitoring of SEP,MEP and DNEP model were used during surgery.All subjects with no neurological deficits preoperatively and got satisfied outcomes.Respectively evaluate the results of neurophysiological intraoperative monitoring (IOM).Data were collected to elevate the efficacy and safety of DNEP monitoring.Results All of 108 patients,15 patients (13.9％,15/108) showed significant changes of neurophysiological parameters,of which 9 cases (60％,9/15) were identified as true positive and 6 cases (40％,6/15) were identified as false positive.During the following-up period,2 patients developed permanent neurological deficit,and 3 patients showed transient neurological deficit who got fully recovered within 6 months after operation.DNEP showed alert in all 5 patients with truepositive alarm,of which 2 patients developed permanent neurological dysfunction and 3 cases showed postoperative short nerve dysfunction that got fully recovery within 6 months after operation.The sensitivity and specificity of SEP+MEP and DNEP were 100％ and 97.98％,100％ and 98.99％,respectively.Conclusion Combining use of MEP+SEP+DNEP monitoring during surgical treatment of spinal deformities presented to be a highly reliable method for the detection and prevention of iatrogenic injury.The results confirmed a high efficacy and safety of DNEP monitoring during spinal surgery.
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Objective To investigate the effects of protein active composition of Scorpio on apoptosis of L1210 tumor cells for the purpose of establishing the quality evaluation method of biological effect of Scorpio. Methods L1210 cells were examined by trypan blue exclusion. The proliferation of cells was determined by improved MTT assay. Flow cytometry was performed to analyze cell apoptosis with propidium iodide (PI). Results When the concentration of protein active composition of Scorpio exceeded or equaled 37 mg/ml, the coefficient correlation of the growth inhibiting curve of L1210 cells was 0.9357, and IC50 was 175 mg/ml. The excellence time was 0 to 48 hours. When the concentration of protein active composition of Scorpio exceeded or equaled 9.25 mg/ml, the apoptosis ratio of L1210 cells was raised significantly.Conclusion The protein active composition of Scorpio might promote the apoptosis and restrain the proliferation of L1210 cells. The value of anti-tumor biological effect of the protein active composition of Scorpio was 9.25 ～ 175 mg/ml. This value may be one of the indexes for quality evaluation of biological effect of Scorpio.