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OBJECTIVE To observe the effects of chlorogenic acid on the activation of macrophage induced by lipopolysaccharide （LPS）， and to explore the role of triggering receptors expressed on myeloid cells-2 （TREM2） in the action. METHODS To find a suitable LPS concentration， the cells were cultured with 1， 10 and 100 ng/mL LPS for 24 h. The level of interleukin 6 （IL-6） in the cell culture supernatant and protein expression of inducible nitric oxide synthase （iNOS） in the cells were detected. To search for a suitable chlorogenic acid concentration， the cells were divided into control group， LPS group and three chlorogenic acid （0.01， 0.1 and 1 μmol/L）+LPS groups. The levels of tumor necrosis factor α （TNF-α） and IL-1β in the cell culture supernatant， the protein expressions of iNOS and TREM2 in the cells and cell viability were detected. To observe the effects of TREM2 in chlorogenic acid alleviating macrophage activation， TREM2-small interfering RNA （TREM2-siRNA） was taken to intervene in TREM2 protein expression. The cells were divided into control group， LPS group， chlorogenic acid+LPS group， TREM2-siRNA+chlorogenic acid+LPS group and SC-siRNA+chlorogenic acid+LPS group. After 24 h incubation， the levels of TNF- α and IL-1β in the cell culture supernatant and protein expressions of TREM2， iNOS and nuclear factor κB p65 （NF-κB p65） in the cells were detected. RESULTS 10 ng/mL LPS promoted IL-6 release and increased iNOS protein expression， and 10 ng/mL LPS was taken in the next experiments. Compared with the LPS group， 0.1 μmol/L chlorogenic acid decreased TNF-α jiaji1981@126.com and IL-1β levels， and down-regulated iNOS expression，meanwhile increased TREM2 expression without effect on cell viability， and 0.1 μmol/L chlorogenic acid was taken in the next experiments. Compared with the control group， the protein expressions of iNOS and NF- κB p65 in the LPS group were significantly increased （P＜0.05）； compared with the LPS group， the protein expressions of iNOS and NF- κB p65 in the chlorogenic acid+LPS group were significantly decreased， the protein expressions of TREM2 was significantly increased （P＜ 0.05）； compared with the chlorogenic acid+LPS group， the protein expressions of iNOS and NF-κB p65 of TREM2-siRNA+ chlorogenic acid+LPS group were significantly increased， the protein expressions of TREM2 was significantly decreased （P＜0.05）. TREM2-siRNA could significantly reverse the above effects of chlorogenic acid， while SC-siRNA did not significantly affect the above anti-inflammatory effects of chlorogenic acid. CONCLUSIONS Chlorogenic acid can inhibit the LPS-induced macrophage activation， and its anti-inflammatory may be mediated by TREM2 protein.
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Objective:To investigate the relationship between the polymorphisms of two SNP loci (rs901823 and rs3736228) in the low density lipoprotein receptor-associated protein 5 (LRP5) gene and glucocorticoids-induced osteoporosis (GIOP) in children.Methods:87 children with GIOP who were treated in Beijing Aiyuhua Women’s and Children’s Hospital and Beijing Children’s Hospital affiliated to Capital Medical University from Jan. 2015 to Dec. 2020 were selected as the research objects, and 100 children with normal bone mass who were treated with corticosteroids in this hospital during the same period were enrolled as the control group. Capillary electrophoresis and fragment analysis (SNaPshot) technology were used to genotype SNP sites rs901823 (T>C) and rs3736228 (C>T) ; Quantitative real-time fluorescence quantitative polymerase chain reaction detection method was employed to determine the relative mRNA of LPR5 gene The amount of expression.Results:For the rs901823 locus of the LRP5 gene, the TT, TC, and CC genotype distribution differences between the GIOP group and the control group were statistically significant ( χ2=14.176, P=0.001) . Compared with the TT genotype, carriers of the TC and CC genotypes had a higher risk of GIOP, with OR values of 3.022 (1.189-6.387) and 5.483 (1.452-20.883) ; For academic significance, OR values were 3.412 (1.795-6.587) and 4.352 (1.215-15.982) . For the rs3736228 locus, the distribution of CC, CT, TT genotypes between the GIOP group and the control group was significantly different ( χ2=9.597, P=0.008) . Compared with CC carriers, CT genotype carriers had a significantly increased risk of GIOP, with an OR value of 5.125 (1.721-16.241) . The result of a dominant model was statistically significant, with an OR value of 4.165 (1.335-14.652) , while for TT there was no statistically significant difference between the carrier and the CC genotype ( P=0.512) , and the results of the recessive model also showed no significant statistical significance ( P=0.887) . There was a statistically significant difference in the frequency distribution of T and C alleles at rs901823 between the GIOP group and the control group ( χ2=17.298, P<0.001) , and the difference in the frequency distribution of C and T alleles at rs3736228 was also statistically significant ( χ2=9.356, P=0.002) . The relative expression level of LRP5 gene mRNA in children with GIOP was 1.34±0.26, which was significantly lower than the expression level of LRP5 gene mRNA in children in the control group of 3.06±0.42 ( t=8.248, P<0.001) . Among children with GIOP, the relative expression of LRP5 gene mRNA in patients with rs901823 locus TT, TC, and CC genotypes was statistically significant ( P<0.001) ; the differences in rs901823 locus CC, CT, TT genotype patients were significant. Pairwise comparison of the relative expression of LRP5 gene mRNA showed that there was no significant difference between the TT group and the CT group ( P>0.05) , but the expression of the CC group was significantly higher than that of the CT group and the TT group ( P<0.05) . Conclusion:The rs901823 and rs3736228 polymorphisms of LRP5 gene are correlated with the occurrence of GIOP and can be used as genetic markers for predicting GIOP in children.
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OBJECTIVE@#To investigate the protective effect of serine hydroxymethyl transferase 2 (SHMT2) against hepatic ischemia-reperfusion injury in mice.@*METHODS@#Sixty C57BL/6 mice were divided equally into sham-operated group, saline adeno-associated virus group (AVV-GFP), and adeno-associated virus silencing group (AAV-SHMT2). The adeno-associated virus and normal saline were injected into the tail vein of the mice 2 weeks before establishment of a 70% ischemia-reperfusion model in the liver. qPCR, Western blotting, immunofluorescence and immunohistochemistry were used to detect the changes of AST/ALT concentration, SHMT2, JNK, NF-κB, caspase-3 and downstream inflammatory factors in the mice, and HE staining was used to observe the pathological damage of the liver tissue in each group; the cell apoptosis in the liver was detected using TUNEL assay.@*RESULTS@#The expression of SHMT2 increased with time after hepatic ischemia-reperfusion and reached the highest level at 24 h (the relative expression was 1.5, < 0.05). At 24 h after hepatic ischemia-reperfusion, the levels of AST/ALT in AAV-SHMT2 group (588/416 U/L) were significantly higher than those in the control group (416/345 U/L) and the empty vector group (387/321 U/L) ( < 0.05). Compared with those in the control group and the empty vector group, the level of SHMT2 was significantly decreased in AAV-SHMT2 group (with a relative expression of 0.24, < 0.05), the levels of p-JNK and p-p65 were significantly increased (relative expression of 0.80 and 0.97, respectively, < 0.05), and the levels TNF-α and IL-1β were consistently elevated (relative expression levels of 1.6 and 1.2, respectively, < 0.05). No significant differences were found in these parameters between the empty vector group and the control group (>0.05).@*CONCLUSIONS@#SHMT2 may alleviate liver cell apoptosis in mice with hepatic ischemia-reperfusion injury by inhibiting the activation of JNK pathway and excessive activation of NF-κB pathway to reduce hepatic damage.
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Animals , Mice , Apoptosis , Liver , Methyltransferases , Mice, Inbred C57BL , NF-kappa B , Reperfusion Injury , SerineABSTRACT
Objective To verify the feasibility of discriminating tissue types in the anterior approach of the cervical spine by the Electrical impedance (EI) and decrease the rate of severe complications.Methods Six New Zealand white rabbits and 6 mini-pigs were performed standard anterior cervical surgery.The esophagus,carotid artery,tracheal cartilage,annular ligaments of trachea,longus colli muscle and anterior longitudinal ligament are classified as the prevertebral group;The cortical bone,cancellous bone,anulus fibrosus and nucleus pulposus are classified as the vertebral group.Once all the tissues were exposed completely,over the frequency range of 200-3 000 kHz (the frequency points were 200 kHz,400 kHz,600 kHz,800 kHz,1 000 kHz,2 000 kHz,3 000 kHz),the in vivo EI of prevertebral group and vertebral group were measured by a probe and a precision inductance-capacitance-resistance (LCR) meter;then the data were analyzed with IBM SPSS Statistics 22 software.At every frequency,Kruskal-Wallis test followed by all pairwise multiple comparisons was applied for all the four groups (the prevertebral group of rabbits,the vertebral group of rabbits,the prevertebral group of mini-pigs,and the vertebral group of mini-pigs),respectively.P values ＜ 0.05 were considered statistically significant.Results At every frequency,the results of Kruskal-Wallis test for all the four groups were significant,and the results of multiple comparisons were as follows.(1) In both miniature experimental animals and large experimental animals,the EI of the vertebral group tissues was significantly different at every investigated frequency (multiple comparisons).(2) In the miniature experimental animals,the EI between longus colli muscle and esophagus,carotid artery and annular ligaments of trachea,and tracheal cartilage and anterior longitudinal ligament had no significant difference at any frequency (multiple comparisons);significant difference could not be detected in the EI between anterior longitudinal ligament and annular ligaments of trachea at the frequencies of 2000-3 000 kHz (multiple comparisonsk) and could be detected at the frequencies of 200-1 000 kHz (multiple comparisons);for comparison of all other paired tissues of prevertebral group there were significant differences at all frequencies (multiple comparisonsk).(3) In the large experimental animals,the EI between longus colli muscle and esophagus,carotid artery and annular ligaments of trachea,and tracheal cartilage and anterior longitudinal ligament had no significant difference at any frequency (multiple comparisons),the same was true for the EI between annular ligaments of trachea and esophagus,carotid artery and anterior longitudinal ligament,and longus colli muscle and annular ligaments of trachea at the frequencies of 800-3 000 kHz,3 000 kHz and 3 000 kHz (multiple comparisons),respectively;for comparison of all other paired tissues of prevertebral group there were significant differences at all frequencies (multiple comparisons).Conclusion At certain frequencies,the EI among tissues was significantly different and could discriminate tissues in the anterior approach of the cervical spine.
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Objective To study the inhibitory effect of curcumin on the proliferation,migration and invasion of non-small cell lung cancer cell A549,and to discuss further if it is closely related to the expression of c-Jun N-terminal kinase (JNK) and relative protein p38.Methods A549 cells were cultured by conventional method,and then treated with different concentration of curcumin (10,20,40,80 μmol · L-1).The proliferation,migration and invasion of A549 cells were measured by real-time cellular analysis (RTCA).The expression levels of JNK,p-JNK,p38 and P-p38 were detected by real-time PCR and Western blotting.Results Curcumin showed an antiproliferation effect against A549 cells with IC50 =40 μmol · L-1,and curcumin exhibited obviously inhibitory effect on the migration and invasion of A549 cells.Additionally,compared with control group,curcumin suppressed the expression of JNK and p38 at the gene level,and significantly inhibited the expression of JNK,P-JNK,p38 and p38 (P＜0.05) at the protein level.Conclusion These results demonstrated that curcumin can inhibit the proliferation,migration and invasion of A549 cells via reducing the level of JNK,p38 phosphorylation,and blocking JNK signal transduction pathway.
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Objective To explore the clinical significance of detecting serum levels of IL-6 and TNF-α in patients with intracranial hemorrhage.Methods The serum levels of IL-6 and TNF-α in different period were detected in patients with intracranial hemorrhage, by enzyme-linked immunosorbent assays (ELISA), and compared with those of healthy subjects (the control group).ResultsThe serum levels of IL-6, TNF-α in the severe and slight patients of study group on 1st, 3rd, 5th and 7th were signiifcantly higher than those in the control group (all withP<0.05). The serum levels of IL-6, TNF-α in the severe patients of study group were signiifcantly higher than those in slight patients of study group (all withP<0.05) on 5th and 7th. The serum levels of IL-6 and TNF-α in dead cases on 5th, 7th days admission were significantly higher than those in survival cases (P<0.05). The serum levels of IL-6 was positively correlated with TNF-α (r=0.721,P<0.05).Conclusion The detection of dynamically serum levels of IL-6 and TNF-α is of great clinical value for assessing the disease development therapeutic efifcacy and prognosis of brain injury patient with intracranial hemorrhage.
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<p><b>OBJECTIVE</b>To evaluate the residual paralysis after a single intubating dose of rocuronium and its effect of residual paralysis after a single dose of rocuronium on the postoperative pulmonary function of patients undergoing laparoscopic gynecological surgeries.</p><p><b>METHODS</b>Sixty American Society of Anesthesiologists (ASA) I - II patients undergoing laparoscopic gynecological surgeries were randomly divided into rocuronium (R) group (n = 30) and rocuronium + neostigmine (R + N) group (n = 30).All patients received midazolam (0.02 mg/kg), fentanyl (1 microg/kg), propofol(1.5-2 mg/kg), and rocuronium (0.6 mg/kg) to facilitate tracheal intubation and no more relaxant thereafter. Anesthesia was maintained with isoflurane and nitrous oxide in oxygen (N(2)O:O(2) = 1:1). At the end of the procedure, neuromuscular blockade was not reversed in R group, while antagonism was accomplished with neostigmine (0.04 mg/kg) and atropine (0.02 mg/kg) in R + N group. Immediately after tracheal extubation and on arrival in the PACU, the train-of-four (TOF) ratio at the adductor pollicis of all patients were measured using acceleromyography. Forced vital capacity (FVC), forced expiratory volume in one second (FEV(1)), and peak expiratory flow rate (PEFR) of all patients were measured using spirometry before surgery, after administration of midazolam and fentanyl, immediately after tracheal extubation, on arrival in the PACU, and after the TOF ratio recovered to 1.0. The TOF ratio and pulmonary function between two groups were compared.</p><p><b>RESULTS</b>Immediately after tracheal extubation and on arrival in the PACU, the mean TOF ratio in R group was significantly lower than that in R + N group (P < 0.05). The mean time to achieve TOF ratio of 0.9 and 1.0 in R group was significantly longer than in R + N group (P < 0.05). Immediately after tracheal extubation and on arrival in the PACU, FVC, FEV(1), and PEFR were significantly lower in R group than in R + N group (P < 0.05). FVC, FEV(1), and PEFR after administration of midazolam and fentanyl and after TOF ratio recovered to 1.0 were significantly lower than the baseline values in all patients (P < 0.01).</p><p><b>CONCLUSIONS</b>After a single intubating dose of rocuronium, residual paralysis exists in the majority of patients undergoing laparoscopic gynecological surgeries. The pulmonary function is impaired after the surgery, even after recovery of TOF ratio to 1.0.</p>
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Adolescent , Adult , Female , Humans , Middle Aged , Young Adult , Airway Extubation , Androstanols , Gynecologic Surgical Procedures , Intubation, Intratracheal , Laparoscopy , Neuromuscular Nondepolarizing Agents , Paralysis , Postoperative Period , Respiratory Function TestsABSTRACT
The purpose of this study was to synthesize the complex of magnetic nanoparticles and antibody, and to apply them to isolate the CD34(+) cells from umbilical cord blood, then to evaluate its separation efficiency. The complex of magnetic nanoparticles and antibody was used to separate CD34(+) cells from umbilical cord blood in the outer magnetic field because of its superparamagnetism, specific identification and function of combination with CD34(+) cells. Scanning electron microscopy was used to observe the morphology of the separated CD34(+) cells. Flow Cytometer was applied to evaluate the sorting efficiency of magnetic nanoparticles, liquid culture and colony culture were taken to assay proliferation and differentiation capacity of the separated CD34(+) cells. The results showed that the CD34(+) cells from umbilical cord blood were isolated fast and effectively by the complex of magnetic nanoparticles and monoclonal antibody. Moreover, the isolated CD34(+) cells still maintained its normal morphology, highly proliferative and differentiative capacity. It is concluded that the complex of magnetic nanoparticles and monoclonal antibody has been successfully synthesized and developed as a technique which efficiently and quickly isolates CD34(+) cells from umbilical cord blood.