OBJECTIVE:To establish an RP-HPLC analysis of Carthamin in mice and to study its pharmacokinetics.METHODS:The serum concentration of Carthamin was determined by RP-HPLC.The blood concentration-time curve was established and the main pharmacokinetic parameters were computed.RESULTS:The linear range of Carthamin was 0.558～55.8 ?g?L-1(r=0.999 2),with the lowest limit of detection at 0.005 ?g?L-1Carthamin in vivo assumed two-compartment model and rapid absorption.CONCLUSION:The proposed method is simple,sensitive and reproducible,and it met the standard for pharmacokinetic study.

Objective To evaluate the clinical significance of cytokeratin19(CK19) mRNA in the peripheral blood of patients with non-small cell lung cancer (NSCLC). Methods The expression of CK19 mRNA was detected by reverse transcription polymerase chain reaction (RT-PCR) in the peripheral blood from 98 NSCLC patients (NSCLC group), 20 benign pulmonary lesion patients (benign pulmonary lesion group) and 20 healthy adults(control group). Results The positive rate of CK19 mRNA in peripheral blood of NSCLC group, benign pulmonary lesion group and control group was 77.55 % (76/98), 3.57% (1/28),0 (0/20) respectively,and there was significant difference among three groups ( x2 = 73.680,P ＜ 0.01 ). The positive rate of CK19 mRNA in different pathological types of NSCLC had no significant difference (x2 =0.414, P ＞ 0.05 ), but had significant difference in different lymphy node metastasis status( x2 = 17.523, P ＜0.01 ) and different clinical stage ( x2 =13.453,P ＜ 0.01 ). Conclusions The higher expression of CK19mRNA in peripheral blood of NSCLC may relate to cancer metastasis. Detection of CK19 mRNA has significance in early prediction of its prognosis.

Objective This study was designed to investigate the importance of H2S in rats with adriamycin-induced dilated cardiomyopathy.Methods Weight-matched adult male Wistar rats were randomly divided into 5 groups as follows:(1)the ADR group(n=12)where adriamycin(2.5 mg/kg)was given once a week;(2)ADR + small-dose NaHS group(n=12)where NaHS solution [2.8 ?mol/(kg?d)] was given;(3)ADR+large-dose NaHS group(n=12)where NaHS solution [14 ?mol/(kg?d)] was given;(4)the control group(n=9)where physiological saline was administered instead of adriamycin.(5)NaHS group(n=9)where NaHS solution [14 ?mol/(kg?d)]was given.Hemodynamic and echocardiographic measurements were carried out after treatment.Histological examination was implemented at the 10th week after the rats were sacrificed.Meanwhile,H2S concentrations in serum and myocardial tissues were evaluated by modified sulfide electrode method.Results The heart function of ADR rats decreased obviously.Pathological examination demonstrated that characteristics consistentwith cardiomyopathy were found in ADR group.H2S concentrations in serum and myocardial tissues of the rats in ADR group were both significantly decreased as compared with those in the control group(both P

ObjectiveTo investigate the effects of ABI-1 gene knockdown upon the proliferation and migration of human gastric cancer cell NCI-N87 in vitro. MethodsNCI-N87-ABI-I-ShRNA cell model was successfully constructed and validated by Real-time PCR and Western blot. The cellular morphous and skeleton, proliferative and migrative potents, and also AKT expression were compared between NCI-N87-ABI-1-ShRNA and its parents by immunofluorental staining, CCK-8 assay, transwell chamber and Western blotting.ResultsCCK-8 assay showed there was no significant difference in the proliferation rates at different time points between the NCI-N87-Vector and NCI-N87 cells while the proliferation rates at the time points of 36 and 48 hours of the NCI-N87-ABI-1-ShRNA were significantly lower than the NCI-N87( t =2. 85and 4. 166, P ＜ 0. 05 ). Transwell assay showed that migrated cell number were 66 ± 8, 65 ± 8 and 30 ± 4,respectively, and there was significant difference between the NCI-N87-ABI-1-ShRNA and NCI-N87 cells (t =9. 550,P ＜0. 05). Finally, ABI-1- knock-down altered the cellular morphoos and skeleton of 90％NCI-N87 cells and inhibited p-AKT expression.ConclusionABI-1 inhibits proliferation and migration of NCI-N87 cells in vitro probably by PI3K/AKT pathway.

Objective To investigate the clinical value of spinal nonfusion interspinous Coflex system for the degenerative lumbar spinal stenosis.Methods From March 2008 to August 2009,26 patients with degenerative lumbar spinal stenosis were treated by decompression with posterior spinous process and interspinous implant Coflex folder method,including 11 males and 15 females,with the mean age of 65.4years(range,45-78 years).L3,4 segment in 7 cases,L4,5 segment in 13 cases,concomitant L3,4,L4,5 segment stenosis in 6 cases.Patients were scanned by MRI and CT confirmed L3,4 and(or)L4,5 segments of the ligamentum flavum thickening,proliferation of small-joint bone,merger disc herniation causing central tube and lateral fossae stricture,nerve root or coccygeal plexus compression.Lumbar anteroposterior,lateral and flexion-extension X-ray films preoperatively,and at follow-up were used to measure the following parameters by eFilm and CAD software:the anterior and posterior disc space height,the range of motion at surgical level,and the segment of the spinal canal area of responsibility.Postoperative standard Japanese Orthopaedic Association(JOA)score for preoperative and postoperative scores.Results Patients were followed up 12-24months.The pain relieved in all patients.The JOA scores improved from(15.46 ±4.30)preoperatively to (24.50±1.58)postoperatively,the vertebral canal area of the affected segment was(218.4±16.2)mm2 before the operation,(264.6±9.9)mm2 after the operation.Single segment anterior disc space height did not change significantly,the difference was not statistically significant.The disc space posterior height increased compared with the preoperative height,and gradually decreased with time.No patients suffered Coflex loosening,fracture and emerge.Conclusion The treatment of lumbar spinal stenosis with Coflex system has satisfactory effect in minimal invasiveness and high security,which provides a safe and effective therapy for degenerative lumbar spinal stenosis.

Objective To investigate the inducing effect of acitrotin on the growth and apoptosis of human cutaneous squamous cell carcinoma cell line SCC13 and on caspases expression.Methods Human cutaneous squa-mous cell carcinoma cell line SCC13 was treated with five different concentrations of acitrefin [5×10-7,1×10-6,5×10-6,1×10-5,5×10-5mol/L].Cell proliferation was evaluated by MTT assay.Apoptosis was assessed by en-zyme-linked immunosorbent assay.The cell cycle was assessed by flow cytometry.The protein expressions of caspase-8 and caspase-9 were examined with Western blot.Results Acitretin inhibited the growth ( F = 83.64,96.34 and 123.17, respectively on the first, third and fifth day)and induced the apoptosis of SCC13 cells(F=74.45,107.37,and 64.28, respectively on the first, third and fifth day) in a dose- and time-dependent manner(P<0.05).Acitretin altered cell cycle distribution of SCC13 cells as compared with controls, the G1-phase population increased by 77.66% 72 hours after acitretin treatment, while the control increased only by 63.55%.An active fragment of caspase-8 occurred following 12 hours treatment of acitretin on SCC13 cells, whereas the caspase-9 active fragment occurred 24 hours after acitretin treatment, which increased time-dependently (P<0.01).Conclusion Acitretin plays an important role on the growth inhibition and apoptosis of cutaneous squamous cell carcinoma cells, which may be affected through both Fas receptor way and mitochondria way.

Objective To investigate the relationship between apolipoprotein E(Apo E) gene poly-morphism and mild cognitive impairment (MCI) in patients with type 2 diabetes mellitus (T2DM), and e-valuate the correlative risk factors. Method 40 cases of type 2 diabetes with MCI and 80 cases of type 2 diabetes without MCI were enrolled in this study. The polymorphism of the Apo E gene was detected by PCR-restriction fragment length polymorphism(PCR-RFLP). According to the clinical data such as course of disease, plasma glucose, plasma fat and body mass index (BMI), the independent risk factors of T2DM and MCI were analyzed by non-conditional logistic regression. Results The frequency of Apo E ε_4 allele in the group of type 2 diabetes with MCI was higher than that without MCI ( 25.0% vs 10. 0% ), and the difference had statistical significance( P < 0. 01 ). The indexes of the statistical significant difference be-twcen the two groups were age, course of disease, postprandial blood glucose ( P2BG), HBA1C, BMI,family history of T2DM, hypertension, diabetic retinopathy, diabetic peripheral neuropathy, Apo E gene. The independent risk factors included diabetic retinopathy ( OR = 3. 452, P < 0. 05 ), diabetic peripheral neuropathy( OR = 3. 252, P <0. 05), Ape E gene( OR = 2. 441, P < 0.01 ), HBA1C ( OR = 1. 372, P <0.05), P2BG(OR = 1. 194, P <0.05), age(OR = 1. 194, P <0.01) and course of disease(OR =1. 142, P <0. 05). Conclusion Apo E ε_4 allele has significant relationship with T2DM and MCI. The age, course of disease, control of plasma glucose, and microvascular complication of diabetes have relation-ship with the cognitive function.

Objective To explore the situation of respiratory virus co-infection with EV71 and CA16 in patients with hand,foot and mouse disease(HFMD) ,and analyze the influence of co-infection on clinical aspects.Methods From June to October of 2010,there were 348 patients enrolled in the study,with 248 hospitalization cases and 100 mild outpatients.All the patients were diagnosed as HFMD in Beijing You-an Hospital.The viral RNA from the pharynx swab samples were extracted and reversely transcribed by RT-PCR.All the samples were detected with the EV71 and CA16 by real-time fluorescence quantitative PCR.Twelve kinds of respiratory viruses were detected by a commercial multiplex-PCR method.The PCR products were confirmed by electrophoresis.Chi square test was used in the data analysis.Results Of the 348 HFMD patients,36 subjects were detected as positive for respiratory virus co-infection.In the 248 hospitalization cases,111 cases were positive for EV71 or CA16,with eight cases identified with respiratory virus co-infection(7.2%); the other 137 cases were negative for EV71 and CA16,with eleven cases identified with respiratory virus co-infection(7.4%).There was not significant difference between respiratory virus co-infection and the identification of EV71 /CA16(x2 = 0.059,P ＞ 0.05).In the 100 mild outpatients positive for EV71 or CA16,seventeen cases were identified with respiratory virus co-infection(17%).The rate of respiratory virus co-infection in the mild outpatients was much higher than in the severe hospitalization patients(x2 = 4.830,P＜ 0.05).Among the 111 EV71(+) or CA16(+) inpatients,there were 101 cases diagnosed as severe cases(91.0%); similarly,there were 132 cases diagnosed as severe cases(96.4%) among the 137 EV71(-) CA16(-) cases.There was not difference between the identification of EV71/ CA 16 and illness of HFMD(x2 = 3.099,P ＞ 0.05).The leading respiratory virus being identified were HRV A/B,PIV3 and FLU A in the 348 HFMD patients.Conclusions Co-infection with respiratory virus exists in the HFMD patients. However,the respiratory virus infection has no significant influence to the state of HFMD illness.

Objective To explore the prognostic value of N-terminal pro-brain natriuretic peptide ( NT-pro-BNP) levels in critically ill infants. Methods Eighty-one critically ill infants were enrolled from January 2013 to January 2014 in pediatric intensive care unit. The minimum of pediatric critical illness score ( PCIS) and the number of dysfunction organs were calculated within 24 hour after admission. According to PCIS,the critically ill infants were divided into extremely critical group(PCIS≤70,n=25),critical group (PCIS 71-80,n=30)and non-critical group(PCIS>80,n=26). According to the prognosis,the critically ill infants were divided into survival group (n=68)and death group(n=13). The serum NT-pro-BNP levels were determined on the first day,third day and convalescent phase. The relationships of serum NT-pro-BNP levels with PCIS and the number of dysfunction organs and prognosis were observed. Results The study showed statistical significances of serum NT-pro-BNP levels among the extremely critical group, critical group and non-critical group,whether on the first day,or on the third day and convalescent phase(P<0. 01). There were statistical significances of serum NT-pro-BNP levels among different stages of the disease in each group(P<0. 01). Compared with survival group,PCIS was significantly lower and the serum NT-pro-BNP levels and the number of dysfunction organs were significantly higher in death group. The serum NT-pro-BNP level on the third day was higher than that on the first day in death group ( P<0. 01 ) , while no significant difference was found in survival group. The serum NT-pro-BNP levels on the first day and the third day and PCIS were negatively correlated(r= -0. 59,P<0. 01;r= -0. 66,P<0. 01). The serum NT-pro-BNP levels on the first day and the third day and the number of dysfunction organs were positively correlated(r=0. 40,P<0. 05;r=0. 57,P<0. 01). Conclusion The serum NT-pro-BNP levels of the critically ill infants are correlated with disease severity,and can be useful for assessing the severity of critical illness.

Objective:To investigate the relationship between tumor necrosis factor-α (TNF-α)-308A/G gene polymorphism and mild cognitive impairment(MCI)in patients with type 2 diabetes mellitus(T2DM),and their correlative risk factors thereof.Methods:Forty cases of T2DM with MCI and 80 cases of T2DM without MCI were selected for this study.The polymorphism of the TNF-α-308A/G was detected by PCR-restriction fragment length polymorphism (PCR-RFLP).According to the clinical data,such as course of disease,plasma glucose,plasma fat and body mass index(BMI),the independent risk factors of T2DM and MCI were analyzed by non-conditional logistic regression.Results:The frequency of TNF-α2 allele was significantly higher in the group of T2DM with MCI than that without MCI (P＜0.01).The indexes of the statistical significant difference between the two groups were the age,course of disease,postprandial blood glucose(P2BG),glycosylated hemoglobin,body mass index,family history of T2DM,hypertension,diabetic retinopathy,diabetic peripheral neuropathy and TNF-α.The independent risk factors included TNF-α,diabetic peripheral neuropathy,diabetic retinopathy,age and P2BG.Conclusion:There is a significant relationship between TNF-α2 allele and T2DM with MCI.There is a significant relationship between the age,control of plasma glucose and microvaseular complication of T2DM with the cognitive funotion.