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Objective:To evaluate the proliferative activity of and changes in the expression of related differentiation proteins in a stably NCSTN gene-silenced human immortalized keratinocyte cell line HaCaT, and to preliminarily explore the possible mechanism underlying the occurrence of acne inversa.Methods:By lentivirus-mediated short hairpin RNA (shRNA) , a NCSTN gene-silenced HaCaT cell model was established (shRNA group) , and other HaCaT cells transfected with empty lentivirus served as a negative control group. Real-time quantitative PCR and Western blot analysis were performed to determine the NCSTN gene-silencing efficiency. Cell counting kit-8 (CCK8) assay was conducted to evaluate the proliferative activity of HaCaT cells, and real-time quantitative PCR and Western blot analysis were performed to determine the mRNA and protein expression of cytokeratins (CK1, CK5, CK7, CK10, CK14, CK16, CK17, CK18, CK19 and CK20) and other differentiation molecules (involucrin and loricrin) respectively in HaCaT cells. Two-independent-sample t test was used to compare the measurement data between two groups. Results:NCSTN mRNA and protein expression were significantly lower in the shRNA group (0.42 ± 0.19, 0.30 ± 0.07 respectively) than in the negative control group (1.00 ± 0.34, 1.00 ± 0.26; t = 5.196, 2.637, P < 0.001, < 0.05, respectively) , and the gene-silencing efficiency was 70%. Compared with the negative control group, the shRNA group showed higher cellular proliferative activity, but decreased protein expression of CK16, CK19 and terminal differentiation molecule involucrin ( t = 3.787, 3.817, 2.904, P < 0.01, < 0.05, < 0.05, respectively) . Conclusion:Stable silencing of NCSTN gene can lead to abnormal proliferation and differentiation of HaCaT cells, which provides new ideas for subsequent exploration of acne inversa caused by NCSTN gene mutation.
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Objective To examine the relationship between the perioperative changes in cytokines and the postoperative complications after esophagecotomy. Methods Twenty-five ASA Ⅱ-Ⅲ patients undergoing esophagectomy were divided into two groups: group A without postoperative complication (n = 17), group B with postoperative complications including pulmonary infection (in 2 patients), pleural effusion (in 3 patients), cardiac arrhythmia(in 2 patients) and anastomosis hemorrage(in 1 patient) (n = 8). Blood samples were taken before anesthesia(T0 ), at skin incision(T1), 2h after chest was opened(T2 ), 60 min after lungs were inflated (T3) and 1,4, 24h after surgery (T4, T5, T6 ) for determination of serum IL-6, IL-8 and IL-10 concentrations. The durations of SIRS, the definition of which was set by American College of Chest Physicians /Society of Critical Care Medicine (ACCP/SCCM), was also recorded. Results The demographic data including age, sex and body weight were comparable between the two groups. There was also no significant difference in preoperative lung function, duration of surgery, blood loss during surgery and duration of unilateral lung ventilation between the two groups. The duration of SIRS was shorter in group A than that in group B. In both groups serum IL-6 and IL-8 levels increased significantly at T2 (after thoracotomy) reached their peak values at T4, and then gradually declined but were still significantly higher than the baseline values(T0). The serum IL-6 level was significantly higher at T6 (24h after surgery) in group B than that in group A. The serum IL-8 level was significantly higher at T3-6 in group B than that in group A. The IL-6/IL-10 and IL-8/IL-10 ratio were significantly lower at T5-6 in group A than those in group B. Conclusions The postoperative complications may occur due to the inflammatory response, and/or anti-inflammatory mediators insufficiency. The IL-6/IL-10 and IL-8/IL-10ratio may be of value in predicting the prognosis.
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Objective To evaluate the effects of propofol and isoflurane on the change in periopreative serum IL 6,IL 8,IL 10 and the balance between pro and anti inflammatory cytokines.Methods Twenty five ASAⅡ Ⅲ patients scheduled for esophagus cancer surgery were randomly allocated to one of the two groups: isoflurane group(group I) and propoful group(group P). All patients were premedicated with oral diazepam 5mg and intramuscular atropine 0.3mg . In group P anesthesia was induced with propofol 1 2 mg?kg -1 ,fentanyl 2 ?g?kg -1 ,midazolam 2 3 mg and vecuronium 0.1mg?kg -1 and maintained with propofol 5 10 mg?kg -1 ?h -1 , inhalation of nirous oxide (N 2O:O 2=50%:50%) and intermittent of boluses of vecuronium. In group I anesthesia was induced with 3% 4% isoflurane ,fentanyl 2?g?kg -1 ,midazolam 2 3mg, vecuronium 0.1mg?kg -1 and maintained with inhalation of 50% N 2O and isoflurane (ended tidal isoflurane was maintained at 0.6%) and intermittent boluses of vecuronium. During operation BP and HR were maintained with at ?20% of pre anesthesia level. After operation all patients received PCEA with 0.15% bupivacaine and fentanyl 2?g?ml -1 . Blood samples were taken from internal jugular vein before anesthesia (T 0), at skin incision(T 1), 2h after thoracotomy (T 2), 60min after lung were inflated(T 3),1,4 and 24h after operation(T 4,5,6 ) for determination of serum IL 6,IL 8 and IL 10 concentrations. Results All patients showed significant increases in IL 6, IL 8 and IL 10 levels during thoracotomy (P