Objective To investigate the effect of etomidate on porcine adrenal cortical cells and the influence of preconditioning with small dose etomidate. Methods Porcine adrenal cortical cells were cultured in Dulbecco's modified Eagle's medium (DMEM) supplemented with 15% fetal bovine serum at 37℃ in 5% CO2 incubator for 24 h. The concentration was 2 × 106/ml. The experiment was performed in 2 parts. In part Ⅰ the cells were exposed to 50, 100, 200, 300, 400, 500 and 1000 mol/L etomidate respectively and incubated for 6, 12and 24 h, while in control group, the cells were exposed to 0.5% dimethylsulfoxide (DMSO). Cell viability was measured using CCK-8 assay and apoptosis by flow cytometry, and the 50% inhibitory concentration (IC50) of etomidate was calculated at 24 h of incubation. In part Ⅱ the cells were exposed to 0.6 μmol/L etomidate for 1 h and were allowed to recover for 4 h after etomidate washout, then etomidate 325 μmol/L was added and the cells were incubated for 24 h. Cell viability and apoptosis were assessed and the IC50 of etomidate was calculated as in part Ⅰ .Results Etomidate inhibited viability of porcine adrenal cortical cells and induced apoptosis in a dose- and time-dependent manner. The IC50 of etomidate at 24 h of incubation was 325 μmol/L. Preconditioning with0.6 μmol/L etomidate for 1 h attenuated the apoptosis induced by etomidate 325 μmol/L. Conclusion Etomidate can inhibit cell viability and induce apoptosis in a dose- and time-dependent manner. Preconditioning with small dose etomidate has protective effect.

With the rapid development of pharmacogenetics, more and more studies have shown evidence in the association between polymorphisms at the human leukocyte antigen (HLA) loci and severe adverse drug reactions (SADRs). Several HLA-B alleles proved to be associated with SADRs for drugs such as carbamazepine, allopurinol, lamotrigine, and lfucloxacillin. hTe USA Food and Drug Administration (FDA) has even recommended routine screening for HLA-B allele before the use of abacavir and carbamazepine. With the completion of human genome project and the Hapmapproject, several new pharmacogenetics approaches such as genome-wide association study (GWAS) have emerged. hTese newly developed methods will undoubtedly accelerate the identiifcation and clinical utilization of the pharmacogenetic biomakers. In addition, the immunogenetic mechanisms by which the HLA alleles cause SADRs are explored at the cellular and molecular level. hTis review focuses on the recent progresses in HLA alleles and ADRs regarding both the clinical translation and modern pharmacogenetic methods.

Objective To investigate the effect of ropivacaine preconditioning on the toxicity of ropivacaine to ND7/23 cells.Methods ND7/23 cells were cultured in DMEM culture medium at 37℃ in 5% CO2 incubator for 24 h at a concentration of 1 × 106/ml. The cells were randomly divided into 3 groups ( n = 3 each) ;control group (group C), ropivacaine group (group R) and ropivacaine preconditioning group (group RP). In group R, the cells were exposed to 1% ropivacaine 100 μl and incubated for 1 h. In group RP, the cells were exposed to 0.02% ropivacaine 100 μl for 15 min, after ropivacaine was washed out, 1% ropivacaine 100 μl was then added and the cells were incubated for 1 h. The cell viability was measured using CCK-8 assay and apoptosis using Annexin-V/PI staining.Results Compared with group C, the cell viability was significantly decreased, while the early apoptotic rate and late apoptotic rate were significantly increased in groups R and RP ( P ＜ 0.05). Compared with group R, the cell viability was significantly increased, while the early apoptotic rate and late apoptotic rate were significantly decreased in group RP (P ＜ 0.05) .Conclusion Ropivacaine preconditioning can protect ND7/23 cells from bupivacaine-induced cytotoxicity through inhibiting apoptosis.

ObjectiveTo investigate the clinical results of hepatectomy for bleeding of spontaneous rupture of hepatocellular carcinoma (HCC). MethodsThe clinical data of 24 cases admitted to our hospital from Jan 1990 to Mar 2004 was analyzed retrospectively.ResultsSurgical hemostasis was achieved successfully in 100.0% (24/24) of patients. The postoperative mortality rate was 4.1% (1/24), and the (complication) rate 12.5%(4/24). Liver function recovered within two weeks after operation. The length of hospital stay was 14.756.25 days.Twenty-one patients were followed-up from 6 month to 36 months. 8 patients died from recurrence within 10 months postoperatively; 10 patients survived over 1 year, 1 patient over 2 years and 1 over 3 years. The overall 1-year survival rate was 58.3%(13/24).ConclusionsIn the management of bleeding of spontaneous rupture of HCC, hepatectomy can effectively stop the bleeding and excise the tumor at the same operation. In some patients, a radical excision can be achieved, and, if the (patient)s condition permits it, hepatectomy should be the treatment of choice.

Objective To explore the clinical significance of detection of plasma microRNA-21,-143 in identifying early esophageal cancer and esophageal non-tumor diseases.Methods The expression of plasma microRNA-21,-143 in 27 cases of patients with early esophageal cancer (esophagus cancer group),25 cases of patients with non-esophageal tumor (non-esophageal tumor group)and in the healthy controls were detected by RT-PCR,and detected the levels of plasma CEA and CA72-4 by the electrochemical luminescence technology,which of changes were analysed to observe the relationship between the changes and the esophageal cancer,the benign esophageal diseases for the two markers.Results The expression of plasma microRNA-21,-143 in the esophagus cancer group were 0.93±0.17,0.27±0.05,which of ones in the non-esophagus cancer group were 0.25±0.03,0.99±0.15,and with those in the control group were 0.23±0.03,1.02±0.15.Compared with those in the non-esophagus cancer group,the expression of plasma microRNA-21,-143 were obviously up or down-regulated with significant differences (t=10.87,11.55,P＜0.01).Compared with those in the control group,which of ones were obviously up or down-regulated with significant differences (t=9.20,9.07,P＜0.01),and with no statistical significances in comparison between the esophagus cancer group and the control group (t=1.39,1.19,P＞0.05).The positiverate of plasma microRNA-21,-143 in the esophageal cancer,non-esophagus cancer group and the control group were,81.4 ％ (22/27),4.0 ％(1/25) and 0 (0/24);85.1％ (23/27),4.0％ (1/25),and 0 (0/24),respectively.The positive rate of microRNA-21,-143 in the esophageal group respectively in comparison with those in the non-esophagus cancer group and the control group were significantly higher,the differences had statistical significances (x2 =31.59,34.39,P＜ 0.01;x2 =34.42,37.23,P＜ 0.01).The expression of two markers in the esophagus cancer group were no statistically significant differences compared with control group (x2 =0.980,0.980,P＞0.05).The sensitivity and specificity of microRNA-21,-143 in early diagnosis on the esophageal cancer were 81.4 ％,97.9 ％ and 85.1％,97.9 ％.The sensitivity of microRNA-21,-143 in the esophageal group were significantly higher compared with those of CEA and CA72-4,the differences were statistically significant (x2 =12.79,P＜0.01;x2 =5.33,P＜0.05;x2 =15.03,P＜0.01;x2 =6.95,P＜0.05).The specificity of microRNA-21,-143 in the esophageal cancer group were no statistically significant differences in comparison with those of CEA and CA72-4 (x2 =1.043,0.000,P＞0.05) and (x2=1.043,0.000,P＞0.05),respectively.The analysis results from the spearman correlation test showed that in the esophageal cancer group,the expression of plasma microRNA-21,-143 had a negative correlation (r =0.658,P＜0.01).Which of ones respectively associated with the levels of CEA and CA72-4 (r=0.607,0.623,P＜0.01 and r=0.579,0.610,P＜0.01).Conclution The detection of expression of plasma miRNA-21,miRNA-143 in the patients with the early esophageal cancer and non-esophageal tumor can provide a new train of thought for pathologic diagnosis of early esophageal cancer.

Objective To explore the clinical application of the expression of LOXL2 mRNA and Tenascin-C mRNA in tissues for the disease with the bile duct cancer.Methods The serum and clinical data in 35 cases of patients with the bile duct cancer (cancer group) and 28 cases of patients with normal bile duct tissue (control group) were collected,used the real-time fluorescent quantitative PCR (real-time-PCR,RT-PCR) technology to detect the expression of LOXL2 mRNA and TenascinC mRNA in tissues toobserve the relationship between the changes and the bile duct cancer for the two markers.Results The expression of LOXL2 mRNA and Tenascin-C mRNA in tissues in the cancer group were 1.27±0.18 and 1.39±0.19,which of ones in the control group were 0.20±0.06 and 0.23±0.06.In the cancer group,the expression of LOXL2 mRNA and Tenascin-C mRNA in tissues respectively with comparision to those in the control group were significantly higher,the differences had statistical significance(t=52.18,56.87,P＜0.01),which of ones in the cancer group was positively related (r=0.687,P＜0.01).Conclution The expression of LOXL2 mRNA and Tenascin-C mRNA in tissues may be a molecular targets for the disease with the bile duct cancer in the early diagnosis and judgment of progression in the courses of this disease.

Objective Macrophage activation syndrome (MAS) is a severe, potentially life-threatening clinical condition. The clinical features including precipitating events, clinical presentations, treatment strategies, outcome in systemic onset juvenile idiopathic arthritis (So-JIA) children with MAS were reviewed. Perforin A91V gene analysis was also performed. Methods Retrospective review of fourteen MAS cases with So-JIA from 2003 to 2008 from a collected database. Gene-specific polymerase chain reaction ( PCR) primers were used to analyze the perforin A91V gene polymorphism. Results Fourteen patients with age from 4 months to 12 years were considered to have evidence of MAS. Nine of them were boys. The primary diagnosis was systemic onset juvenile idiopathic arthritis. No medication was identified as trigger. Eleven of them had infections prior to MAS. Among them specific infectious agents were identified in four patients. High fever, new onset of hepatosplenomegaly, lymphadenopathy, liver function abnormality, abnormal lipid metabolism and hemophagocytosis were common clinical features. Two cases presented with acute respiratory distress syndrome (ARDS). Multiple organ failure (MOF) occurred in three cases. Three patients died. The variant form (NCBI: SNP rs35947132) of perforin A91V gene was detected in seven systemic onset juvenile idiopathic arthritis compolicated with MAS cases. However no mutation was detected. Clucocorticoid, intravenous immunoglobulin, immunoimpressive therapy were effective treatment of this condition. Plasmapheresis (HP) was successfully used in one case with severe MAS. Conclusions MAS is a rare and potentially fatal complication of childhood rheumatoid diseases such as systemic onset juvenile idiopathic arthritis. In this series, majority of them were male and most of them were preceded by infection. Bone marrow studies support the diagnosis. MOF may be a poor prognostic sign of So-JIA. Aggressive and early therapy is essential. There is no relationship between the variant form (NCBI: SNP rs35947132) of perforin A91V gene and So-JIA with MAS in this small sample's study. More research need to be done by increasing sample's numbers.

Objective To study effect of nasal tolerance with rat-derived 97-116 peptide of AChR α-subunit(Rα97-116(V108A))on the manifestation of muscle weakness and the immunity function of experimental autoimmune myasthenia gravis(EAMG).Methods Twenty-two EAMG model Lewis rats immunized thrice with Rα97-116(V108A)were divided randomly into tolerance group and control group.They were respectively immunized with Rα97-116(V108A)and PBS buffer solution for 10 days via nasal mucous.Then the body weight and Lennon score of two group Lewis rats were measured.Their serum anti-AChR antibodies were tested by ELISA,the expression levels of CD28,CTLA4,B7-1 and B7-2 were determined by flow cytometry.Results Compared with control group at different time points.the body weight of tolerance group rats(tolerance group(228.1±5.8)g,control group(215.0±16.2)g,t=2.395,P＜0.05)increased,the mean clinical score of rats(tolerance group 1.55±0.44.control group 2.10±0.66,t=-2.20,P＜0.05)decreased and the amount of serum anti-AChR antibody(tolerance group 0.97±0.20,control group 1.27±0.26,t=-2.857,P＜0.05)decreased obviously.the amount of CD28,B7-1,B7-2,CTLA4(%)expressed on the surface of peripheral blood cells(tolerance group:27.35±7.05,4.73±0.58,2.71±0.35,1.72±0.44,control group:40.02±8.81,9.52±1.25,5.88±1.09,2.64±0.47)down-regulated markedly(t=3.479,10.861,8.755,4.403,all P＜0.01).Conclusion Nasal mucous tolerance with Rα97-116(V108A)could ameliorate muscular weakness of EAMG rats while activates T cell and inhibits B cellular immunity.

Background and purpose:The transcription factor PAX6 is primarily expressed in embryos. PAX6 is also expressed in several tumors and plays oncogenic or tumor suppressor role. This study aimed to investigate the expression of PAX 6 in non-small cell lung cancer (NSCLC) tumor samples and cell lines; and evaluate the effects of PAX6 on the proliferation and invasion of tumor cells and the expression level of cyclin E and p38.Methods:Western blot was carried out to detect the PAX6 protein level in 86 NSCLC tumor tissues, paired adjacent normal tissues and 2 cell lines. PAX6 siRNA was transfected into human lung cancer A549 cell line. Anchorage-independent growth and invasiveness of tumor cells were measured by MTT and transwell cell invasion assay, respectively. Cyclin E and p38 protein level before and after transfection were detected by western blot.Results:Comparison with tumor adjacent tissues and normal human bronchial epithelial cells 16HBE, PAX6 in NSCLC tissues and A549 cell lines was signiifcantly higher expression. After transfection with efifcient sequence of PAX6 siRNA for A549 cell lines, the down expression of PAX6 inhibits tumor cell proliferation, the proportion of cells in G1 phase increased, the cell invasion and migration decreased remarkably. Cyclin E and p38 activity was inhibited inPAX6 knockdown cells.Conclusion:PAX6 accelerate cell cycle progression by activating cyclin E and p38. PAX6 is a potential target for diagnosis and therapy.

Objective: To analyze the risk factors on acute respiratory dysfunction caused death in patients after type A aortic dissection surgery.
Methods: A total of 223 patients who received aorta replacement surgery in our hospital from 2010-01 to 2012-12 were retrospectively studied. 80 patients suffered from post-operative acute respiratory dysfunction including 61 male and 19 female with the mean age of (49.2 ± 11.6) years. Those patients were divided into 2 groups as Death group, n=18 and Survival group, n=62. We analyzed the most relevant risk factors for death, such as gender, age, histories of smoking, diabetes, hypertension, Marfan syndrome;pre-operative acute or chronic dissection, hypoxemia, mal-perfusion, LVEDD and LVEF;CPB time, aortic-clamping time;post-operative ICU retention time, mechanical ventilation time, permanent neurologic dysfunction, pulmonary infection, MACE, renal failure, hypohepatia, septicemia and wound mal-healing, et al.
Results: The early post-operative (< 3 days) respiratory dysfunction rate was 35.8% and the mortality was 22.5%(18/80). The relevant risk factors of death included female gender (P=0.019), haemorrhage (P<0.01), mechanical ventilation time (P=0.011), permanent neurologic dysfunction (P=0.013), pulmonary infection (P=0.001), MACE (P=0.022), renal failure (P<0.01), hypohepatia (P<0.01) and septicemia (P=0.001). Female gender and renal failure were the independent risk factors for respiratory dysfunction caused death in patients after type A aortic dissection surgery.
Conclusion: The occurrence and mortality were high in patients after type A aortic dissection surgery especially in those with female gender and post-operative renal failure.