Aim To develop a sensitive, rapid and ac-curate LC-MSn method for determination of midazolam/1′-hydroxymidazolam and their pharmacokinetic char-acteristics in rat brain. Methods SD rats received in-travenous injection of midazolam ( 5 mg · kg-1 ) via femoral vein, a probe drug of cytochrome P450 3A. The microdialysis ( MD ) samples in situ brain were collected every 8 mins at 2. 0μl·min-1 in 2. 4 hours. Analytes in brain dialysate were quantified by the pro-posed LC-MSn method. Gradient elution was performed on an Agilent Eclipse Plus-C18 column ( 2 . 1 × 50 mm, 3. 5 μm). The mobile phase consisted of 2 mmol ·L-1 ammonium acetate and acetonitrile. The analyte was detected using electrospray ionization ( ESI ) in multiple reaction monitoring ( MRM) modes. The reac-tion selected ions were 326 . 1/291 . 1 m/z for midazo-lam, 342. 1/324. 1 m/z for 1′-hydroxymidazolam and 285. 1/154. 0 m/z for diazepam as internal standard. Result The linear ranges of midazolam and 1′-hydroxymidazolam were 0 . 78~100 and 0 . 195 ~12 . 5μg·L-1 respectively. The lower limit of quantification was 0 . 2 μg · L-1 . The RSD of intra- and inter-batch precisions was less than 7 %. The RSD of accuracy was from -1 . 34 to -8 %. Conclusion This sensi-tive and rapid LC-MSn method is suitable for determi-nation of midazolam/1′-hydroxymidazolam in rat brain dialysate. MD combined with LC-MSn method may give assistance to deep and further studies of drug metabo-lism and CYP3A enzyme in brain.

Objective To explore whether the patients aged over 65 years with obstructive sleep apnea hypopnea syndrome (OSAHS) are able to tolerate continuous positive airway pressure (CPAP)while compared with patients aged less than 65 years.And to investigate the factors that affect compliance to CPAP in OSAHS patients.Methods A total of 147 OSAHS patients diagnosed with overnight polysomnography (PSG) were divided into two groups:≥65 years old group (n=46),<65 years old group (n = 101 ).Clinical data and PSG parameters were included in a computerized database.The pressure level of CPAP,the duration of respirator use were followed up.Results Compared with <65 years old group,there were higher prevalences of COPD (16% vs.4%,P=0.02),cardiovascular disease (23% vs.10%,P=0.04),apnea (43% vs.26%,P=0.03),regular alcohol consumption (61% vs.38%,P=0.007) and lower incidence of snoring (31% vs.54%,P= 0.03),shorter total sleeping time (378 min vs.423 min,P=0.001),longer wake after sleep onset periods (162 min vs.115 min,P=0.004),lower sleep efficiency (69% vs.77%,P<0.001),higher percentage of stage 1 sleep (29% vs.20%,P=0.001),lower percentage of stage 3-4 sleep (6% vs.9%,P=0.016) and rapid eye movement (REM) sleep (12% vs.15%,P=0.001) in ≥65 years old group.The percentages of acceptance to CPAP at 3 months,6 months,1 year,2 years and 3 years were 91%,89%,84%,82% and 82% respectively in ≥65 years old group,and were 92%,86%,81%,72% and 67% in <65 years old group.Average use time of CPAP were (5.1±1.5) h/night in ≥65 years old group,and were (3.9±1.2) h/night in <65 years old group (P=0.022).By analysis of multivariate logistic regression,a high pressure CPAP was associated with higher objective CPAP compliance in OSAHS patients.Conclusions Compliance to CPAP in older OSAHS patients is not decreased when compared to younger adults.A high CPAP pressure is the only significant independent predictor of better CPAP compliance.

Objective To investigate the relationship between polymorphisms of angiotensinogen(AGT) gene T704C and cerebrovascular disease(CVD).Methods AGT T704C genotype and allele were examined by PCR-RLFP in 82 patients with cerebral infarction(CI group),70 patients with intracerebral hemorrhage(ICH group) and 89 age-matched normal controls(NC group).The AGT T704C genotype and allele frequencies among the 3 groups were compared and analyzed.The effectes of AGT T704C genotype and the risk factors of stroke to induce CI snd ICH were analyzed by Logistic regression. Results The frequencies of AGT 704CC genotype and C allelic in CI group(63.4%,79.9%) were significantly higher than those in NC group(34.8%,61.2%)(allP

Objective To compare the clinical efficacy of clopidogrel and clopidogrel in the treatment of myocardial ischemia in patients with coronary heart disease (CHD).Methods86 patients with myocardial ischemia of coronary heart disease from January 2016 to January 2017 in our hospital, were randomly divided into group A (n=43) and group B (n=43), group A was treated with ticagrelor treatment, group B was treated with clopidogrel treatment, clinical curative effect were compared between the two groups.ResultsThe total effective rate in group A was 86.04% was significantly higher than 72.09% in group B, the difference was statistically significant (P<0.05);The number of ST segment descent in group A was (36.17±4.22) times,ST segment continuous decline time (120.29 ±13.53) s ST continued to decline time (120.29±13.53) s, heart rate (65.27±3.18) times/min, systolic blood pressure (105.44±2.17) mmHg, diastolic blood pressure (63.11±3.14) mmHg were significantly lower than the control group (P<0.05);group A patients with triglyceride,total cholesterol,plasma viscosity trimming rate, whole blood viscosity at low shear rate, and whole blood high shear rate were significantly lower than those in group B (P<0.05).ConclusionCompared with clopidogrel, Grillo can effectively improve the blood microcirculation of myocardial ischemia in patients with coronary heart disease, coronary heart disease,relieve myocardial ischemia caused by coronary heart disease The clinical efficacy is remarkable, and it has the value of popularization and Application.

Objective To investigate the personality patterns, coping styles and social support statue of the patients with stroke.Methods 68 patients of stroke and 68 normal controls were measured with Eysenck Personality Questionnaire (EPQ)、Coping Style Questionnaire (CSQ) and Social Support Scale (SSS).Results The scores of E and N(11.98?4.79,12.98?4.73) were very higher in stroke group than those of in normal control group(9.46?4.18,9.60?4.70)(all P

Clinical data management,a critical part of clinical trials,plays a decisive role in assuring the accuracy and reliability of the trail results.This article discusses the quality standards of clinical data,introduces the important rules and principles for standardized clinical data management,and reviews the development and current status of clinical data management home and abroad.It is proposed that we should learn the advanced clinical data management modes from international partners and introduce standardized clinical management software from abroad,so as to improve the clinical data management in China.

Objective To investigate the relationship between renin (REN) gene G10631A, angiotensinogen (AGT) gene T704C mononucleotide polymorphisms and cerebral infarction and to investigate the mechanisms and characteristics of cerebral infarction from molecular level. Methods REN gene G1063A and AGT gene T704C polymorphisms in 82 patients with cerebral infarction and 89 controls were detected with polymerase chain reactionrestriction fragment length polymorphism. The differences of the genotypes and allele frequencies were compared between the patient group and the control group. Results The frequency of REN 10631AA genotype (31. 7% vs. 10. 1%,χ2 =12. 816, P = 0. 002) and the frequency of A genotype (49. 4% vs. 30. 3% χ2 = 12. 969, P =0. 000), as well as the frequency of AGT 704 CC genotype (63. 4% vs. 34. 8% χ2 = 15. 029, P = 0. 001) and the frequency of A genotype (79. 9% vs. 61. 2% χ2 = 14. 173, P = 0. 000) in the cerebral infarction group were all significantly higher than those in the control group; the frequency of haplotype 704C 10631A was also significantly higher than that in the control group (P=0. 000). Conclusions REN 10631AA genetype and A allele as well as AGT 704 CC genetype and C allele may be the susceptible factors of cerebral infarction. Haplotype 704C-10631 A may be a genetic risk factor for the occurrence of cerebral infarction.

OBJECTIVE:To observe the effects and side effects of recombinant erytropoietin(rEPO)on patients with anemia in chronic renal failure(CRF).METHODS:128 patients with anemia in CRF had been given rEPO by subcutaneous injection for12weeks,the clinical effects were observed by own control method.RESULTS:Excellence cases amounted 91,efficacy cases33,and the overall efficacy rate was96.88%;The side effects included hypertension,coagulation in dialysis machine,pain in injection site and head,no severe adverse drug reactions were found;No degradation in renal function was found in non-dialysis patients during the medication.CONCLUSIONS:rEPO could improve anemia in CRF safely and effectively.

With the constant deepening of the study in the genetic factors of eardio-cerebrovascular diseases, the relation between the renin-angiotensin system (RAS) gene polymorphisms and hypertension is increasingly receiving attention. As an important component of RAS, renin has received much concern in the genetic research of cardio-cerebrovascular diseases and its gene polymorphisrns have become the candidate genes of hypertension, coronary heart disease and stroke, etc.

Background and purpose:Hepatitis B virus X protein (HBx) and hypoxia inducible factor-1α(HIF-1α) play key roles in hepatocarcinogenesis and the development of hepatocellular carcinoma. Positive correlation on the expression of these 2 proteins in hepatocellular carcinoma tissues has been found, whereas the underlying mechanisms have not been fully elucidated. This study focused on the role of HBx in regulating HIF-1α and the underlying mechanisms in hepatocellular carcinoma cells. Methods:The expression plasmids were transfected into Huh7 cells with LipofectemineTM 2000. Western blot analysis was applied to detect the expressions of HIF-1αand HIF-1β protein. The transcriptional activity of HIF-1α was detected by the commercial analysis kits. The mRNA levels of HIF-1αand its target genes, including vascular endothelial growth factor (VEGF) and multi-drug resistance gene 1 (MDR1), were detected by quantitative real-time PCR (qRT-PCR). Immunoprecipitation analysis was applied to detect the interaction of HIF-1α, HBx and protein von Hippel-Lindau (pVHL). Results:Huh7 cells transfected with HBx plasmid led to sharp increase of HIF-1αprotein and transcriptional activity, as well as the mRNA of VEGF and MDR1 (P<0.05). However, the mRNA level of HIF-1αwas not obviously changed after HBx transfection (P>0.05). Meanwhile, HBx also signiifcantly impaired the function of pVHL in mediating the degradation of HIF-1αby ubiquitin hydrolase. This finding was further confirmed by the immunoprecipitation analysis, which showed that HBx could directly bind to pVHL, but not to HIF-1α. Conclusion:HBx may inhibit the inter-activation between pVHL and HIF-1αthrough directly binding to pVHL, and thus enhance the stability and transcriptional activity of HIF-1α.