Objective To evaluate the potential of S100 calcium binding proteins S100A2 and S100A6 levels in serum as diagnostic markers for non-small cell lung cancer (NSCLC). Methods Enzyme-linked immunosorbent assay (ELISA) was performed to detect the levels of S100A2 and S100A6 in 141 NSCLC patients and 150 healthy subjects. Results The average level of serum S100A2 in NSCLC group was (15.02±0.79) ng/ml, that in healthy control group was (11.18±0.64) ng/ml, and the difference was statistically significant (P=0.002). The average level of serum S100A6 in NSCLC group was (12 760±651.8) pg/ml, that in healthy control group was (8 434±408.2) pg/ml, and the difference was statistically significant (P<0.001). The serum levels of S100A2 and S100A6 in stageⅠ/ⅡNSCLC patients were higher than those in healthy controls (P<0.05), respectively. Receiver operating characteristic (ROC) curve analysis showed that S100A2 and S100A6 could distinguish NSCLC patients from healthy controls [area under the curve (AUC) were 0.646 and 0.668, respectively]. Meanwhile, these two proteins showed notable capabilities for distinguishing stage Ⅰ/ⅡNSCLC from healthy controls (AUC were 0.708 and 0.702, respectively). Conclusion Serum levels of S100A2 and S100A6 are significantly elevated in the early stage for NSCLC patients, which can be the potential biomarkers for the diagnosis of NSCLC.

Objective To evaluate the inhibition effects of shuanghuanglian injection powder and its active components on the activities of CYP1A,CYP 2D and CYP 3A in rats liver microsomes. Methods Three probe substrates including phenacetin for CYP1A,dextromethorphan for CYP2D and midazolam for CYP3A were incubated with shuanghuanglian injection powder and the active components (baicalin,phillyrin,forsythiaside A,lutin,chlorogenic acid,coffeic acid and lutiolin) in rat liver microsomes. Contents of three probe substrates were simultaneously determined by HPLC to evaluate the metabolic rates. Results Shuanghuanglian injection powder and baicalin inhibited the activities of CYP2D and CYP 3A, but didn 't affect CYP1A. The other active components showed no effect on CYP1A,CYP2D and CYP3A. Conclusion Drug-drug interactions may occur when combining shuanghuanglian powder injection with CYP2D and CYP 3A substrates and baicalin may be the effector substance responsible for the interactions.

Objective To observe the effect of polydatin on nerve cell apoptosis and the influence of the caspase-8 and FLIP protein expression after ischemia reperfusion injury. Methods Thirty rats were randomly divided into sham operation group, model group and polydatin group. The middle cerebral artery occlusion (MCAO) model of focal cerebral ischemia was established by the method of thread embolism. Rats were subjected to adaptive feeding for the first 7 days and then recieved the treatment for another 10 days. The model of ischemia reperfusion injury was established at the 14th day. The polydatin group received 15 mg/kg of polydatin, and the sham operation group and the model group with the same volume of saline once a day for 15 days. The expression of caspase-8 and FLIP protein in the hippocampus of rats were observed by immunohistochemical staining. The expression of caspase-8 and FLIP protein in the hippocampus of rats were observed by TUNEL method at 72 h after cerebral ischemia-reperfusion. Results Compared with the model group, the number of apoptotic cells of polydatin group significantly decreased in hippocampus CA1 region (P<0.01); The expression of caspase-8 (148.78 ± 6.82 vs. 89.61 ± 7.76) in the polydatin group significantly decreased and the expression of FLIP (127.60 ± 8.52 vs. 150.22 ± 8.53) in the polydatin group was increased significantly in hippocampus CA1 region(P<0.01). Conclusions Polydatin have a protection effect on ischemia reperfusion injury in rats and its mechanism may be inhibition of caspase-8 protein expression, promote the FLIP protein expression.

Objective To study the chemical constituents of Clematis manshurica. MethodsThe extract from the roots and rhizomes of C. manshurica was subjected to chromatography on silica gel and Sephadex LH-20 column. The compounds obtained were identified on the basis of their physicochemical and spectroscopic evidences. ResultsEleven compounds were isolated and their structures were elucidated as squalene (Ⅰ), friedelin (Ⅱ), hexacosoic acid (Ⅲ), ?-sitosterol (Ⅳ), stigmasterol (Ⅴ), oleanolic acid (Ⅵ), ?-daucosterol (Ⅶ), 5-hydroxymethyl-2-furaldehyde (Ⅷ), methyl 3, 4-dihydroxy-phenyl lactate (Ⅸ), 5R-dihydro-5-hydroxymethyl-2(3H)-furanone (Ⅹ), 5R-5-hydroxymethyl-2(5H)-furanone (Ⅺ). ConclusionAll the compounds except for ?-sitosterol are isolated from the plant for the first time.

Objective To summarize the clinical characteristics and causes of ischemic cerebrovascular disease(ICD)in children.Methods A retrospective analysis was conducted on the clinical data of 53 cases with ICD from Feb.2002 to Jun.2008 at the department of neurology in Wuhan Children's Hospital.The self-designed questionnaire of children with ICD was used,whose items included patients' age,gender,personal history,clinical features,cerebrospinal fluid examination,neurological imaging,immunologic examination,metabolic examination,and so on.Results Of 53 children with ICD,30 cases(56.6%)were male,and 23 cases(43.4%)were female.Patients' age varied from 9 months to 12 years old,in which 45 cases(84.9%)were less than 6 years old.Patients from rural area(60.4%)were more than those from city(39.6%).Ratio of limb paralysis was 75.5%(40 cases)in first clinical symptomatology of children with ICD,including hemiplegia in 32 cases(60.4%),alternate hemiplegia in 5 cases(9.4%)and monoplegia in 3 cases(5.7%).Skull CT/MRI scan was performed to reveal 27 cases(50.9%)with basal ganglia region infarction and secondly 15 cases(28.3%)with multi-lobar infarction.Forty cases were found in abnormal cerebrovascular image by means of magnetic resonance angiography/digital subtraction angiography,in which middle cerebral artery and its branches were involved in 21 cases(52.5%).There were 41 cases(77.4%)of patients to be found with clear causes,of which 13 cases(24.5%)were of infections,8 cases(15.1%)of moyamoya disease,5 cases(9.4%)of cerebral vascular malformations,4 cases(7.5%)of head trauma.However,another 12 cases(22.6%)of patients had unknown etiology.Conclusions Children with ICD had characteristics themselves.The limb paralysis was mostly the first symptoms,and the middle cerebral artery and its branches lesions were the most common locations in children with ICD,and next the internal carotid artery involvement,anterior cerebral artery involvement,posterior cerebral artery involvement,cerebral vascular malformations,and so on.Their major cause was infection,followed by Moyamoya disease,cerebrovascular malformations and head trauma,and there were still some unknown causes.

Objective To assess the value of ultrasound examination in transvaginal diagnosis of normal fetal heart with 11-14 weeks of gestation.Methods Totally 158 cases of normal fetal heart with high risk pregnancy and nuchal translucency thickness were examined by two ultrasoud approaches:transvaginal(TVS) and tranabdomina(TAS) in 11-14 weeks of gestation.Results The group of TVS was obviously clearer than TAS by displaying the normal fetal cardiac structural in 12 week of gestation and 13-14 weeks of gestation with significant difference between the two groups (P<0.05),respectively.No significant difference between the two groups in 11 week of gestation was observed.Conclusion The transvaginal echocardiogram is of clinical value in the high risk gravida during the late first and the early second trimester.

Objective To evaluate the effectiveness and safety of gefitinib retreatment beyond progression(GRBP)in non-small cell lung cancer(NSCLC)patients with rare EGFR mutations.Methods We retrospectively analyzed six rare-EGFR-mutation NSCLC patients from Jan 2011 to Dec 2015.Those patients had previous disease control and then disease progression according to Response Evaluation Criteria in Solid Tumors version 1.1(RECIST v1.1)after taking oral gefitinib 250 mg once a day.After that,continuing gefitinib was decided by clinicians′ experience at the same treatment option.The primary endpoints were response rate(RR),overall survival(OS),the first and second progression-free survival(PFS-1 and PFS-2).Safety was assessed according to the NCI-CTCAE version 4.0.Results After initial treatment of gefitinib,4 patients achieved partial response(PR)and 2 patients showed stable disease(SD),with RR being 66.7％.The median PFS-1 and PFS-2 were 10 months(95％CI 6.6-13.4)and 9 months(95％CI 6.9-11.1),respectively.The median OS time was 28 months(95％CI 10.4-45.6).The most common treatment-related adverse events were fatigue,diarrhea,rash,itching and elevated transaminases.Conclusion In our study,gefitinib retreatment beyond disease progression is effective with a manageable tolerability profile.

The means of survival of genomic dsRNA of reoviruses from dsRNA-triggered and Dicer-initiated RNAi pathway remains to be defined.The present study aimed to investigate the effect of Grass carp reovirus (GCRV) replication on the RNAi pathway of grass carp kidney cells (CIK).The dsRNA-triggered RNAi pathway was demonstrated unimpaired in CIK cells through RNAi assay.GCRV-specific siRNA was generated in CIK cells transfected with purified GCRV genomic dsRNA in Northern blot analysis; while in GCRV-infected CIK cells,no GCRV-specific siRNA could be detected.Infection and transfection experiments further indicated that replication of GCRV correlated with the increased transcription level of the Dicer gene and functional inhibition of in vitro synthesized egfp-siRNA in silencing the EGFP reporter gene.These data demonstrated that although only the genomic dsRNA of GCRV was sensitive to the cellular RNAi pathway,unidentified RNAi suppressor protein(s) might contribute to the survival of the viral genome and efficient viral replication.

Objective To evaluate the performance of actinic keratosis (AK) tissue as a culture model for the study of interference in transduction pathway,and to explore the mechanism underlying the p53 regulation though TGFβ1/Smads pathway by using the tissue culture model.Methods Twenty-five skin samples from the lesions of patients with AK were cultured,and divided into 5 groups to be treated with TGFβ1 of 10 μg/L for 24 and 48 hours,the tran sforming growth factor (TGF) β1 receptor kinase inhibitor SB431542 of 10 μmol/L for 24 and 48 hours,respectively,or remain untreated.Real time PCR and Western blot were performed to quantify the mRNA expression of p53 and protein expression of p53 and phosphorylated Smad2 in these tissue specimens respectively.Results A significant elevation was observed in the expressions of p53 mRNA ( 13.4968 ± 0.9903 vs.1,P ＜ 0.05) and phosphorylated Smad2 (0.700 ± 0.023 vs.1,P ＜ 0.05) in AK tissues after treatment with TGFβ1 for 24 hours,and in the expressions of p53 mRNA (13.3882 ± 1.6772 vs.1,P ＜ 0.05) and protein (1.009 ± 0.001 vs.0.512 ± 0.005,P ＜ 0.05) after treatment with TGFβ1 for 48 hours,compared with the untreated AK tissues.No significant differences were observed in the expression of p53 protein between the AK tissues treated with TGFβ1 for 24 hours and 48 hours (P ＞ 0.05).SB431542 induced a statistical reduction in the level of phosphorylated Smad2 at 48 hours (0.116 ± 0.003 vs.0.306 ± 0.023,P ＜ 0.05),but no significant changes were observed in the expression of p53 mRNA or protein after SB431542 treatment for 24 or 48 hours.Conclusions AK tissue cultures can serve as a model for the study of interference in signal transduction pathway.TGFβ1 might regulate the expression of p53 protien through Smads pathway in AK.

Objective To compare the efficacy and the side-effect of three different ways in treating the patients with malignant pleural effusion. Methods 98 patients histologically proved malignant pleural effusion were randomly divided into three groups, bleomycin group(BLM), bleomycin with mycobacterium group( BLM + UTL) and blemycin with intertleukino2 ( BLM +IL). 31 patients were treated with bleomycin intrapleural injection in BLM group,32 patients were treated with bleomycin and Utilin's(mycobacterium) intrapleural injection in BLM + ULL group and 35 patients were treated with bleomycin and intertleukin-2 intrapleural injection in BLM + IL group. The therapeutic efficacy, change of performance and side effects were compared among the three groups after one period of treatment. The changes of CEA and TNF in the pleural effusion were examined before and after treatment. Results The therapeutic efficacy and performance improvement were higher in BLM+UTL and BLM+IL group than that of BLM group(P＜0. 05) ,the pleural CEA of post-treatment in three groups were lower than that of pre-treatment(P＜0.01) ,the CEA after treatment in BLM+UTL group and BLM+IL group was lower than that of BLM group(P＜0. 01,respectively). The pleural TNF of post-treatment in BLM+UTL and BLM+IL groups was higher than that of pre-treatment(P＜0. 01 ) in BLM group. The pleural TNF of post-treatment in BLM+UTL and BLM+IL group was higher than that of BLM group ( P＜0. 01 ). Conclusion Intrapleural injection of mycobacterium with bleomycin or interlekin-2 with bleomycin has better efficacy than using bleomycin only in treating malignant pleural effusion.