OBJECTIVE:A RP-HPLC method was established to determine the concentration of ambroxol hydrochloride in human plasma.METHODS:Waters HPLC instrument was used with the Hypersil BDS C 18 column(5?m,250mm?4.6mm). Ditiazem hydrochloride was chosen as the internal standard.The mobile phase was composed of acetonitrile-methyl alcohol-0.01mol/L phosphatic buffer(pH7.0)-tetrahydrofuran(35∶35∶27.5∶2.5).Flow rate was1.0ml/min.Detection wavelength was242nm.A single oral dose of90mg imported ambroxol hydrochloride tablet was given to9healthy male volunteers.Ambro xol concentration in plasma was assayed.RESULTS:The standard curve of ambroxol hydrochloride was linear in the range of10～640ng/ml.The minium detection concentration was10ng/ml.The extraction recovery was more than90%.A one-compartment open pharmacokinetic model was adopted in ambroxol plasma concentration-time data analysis.The main pharmac okinetic parameters were as follows:C max =(226.53?34.59)ng/ml,T max =(1.82?0.80)h,T 1/2ke =(6.27?0.66)h,AUC 0～24 =(2363.55?448.86)ng/(h?ml).CONCLUSION:The method is simple.The sensitivity and accuracy are high.It is applicable to study the pharmacokinetics of ambroxol hydrochloride in healthy male volunteers.

Objective To investigate the influence of benzopyrene on extracellular matrix(ECM) protein deposition of human airway smooth muscle cells(HASMC) and the related pathway mechanism.Methods HASMC were primarily cultured and the 2-6 generations cells were applied in this experiment.The expression amount of ECM gene and protein was detected by real time PCR and Western Blot the phosphorylation level was analyzed by using the Western Blot method.Results Benzopyrene could to increase the expression of HASMC collagen Ⅰ α1 protein (P＜0.01) and ECM protein (including collagen Ⅰ α1,versican,fibronectin,laminin α2) mRNA(P＜0.05).Benzopyrene could induce the rapid increase of ERK1/2 phosphorylation level (P＜0.01).Furthermore,the ERK pathway inhibitor PD98059 could significantly inhibit the increase of benzopyrene-induced collagen Ⅰ α1 (P＜0.01)and ECM protein(including collagen Ⅰ α1,versican,fibronectin,laminin α2) mRNA expression(P＜0.01).Conclusion Benzopyrene induces the ECM protein deposition of HASMC by activating the ERK1/2 pathway,blocking the ERK1/2 signal pathway can inhibit the benzopyrene-induced airway remodeling.

Objective To observe the expression of uPA, tPA and PAI-1 in whole blood of rat membranous ne-phropathy ( MN) models induced by cationic bovine serum albumin ( C-BSA) , and to explore the effect of fibrinolytic sys-tem on podocyte apoptosis and pathological changes. To explore the possible preventive and therapeutic effects and the pos-sible mechanisms of early prevention of fibrinolysis. Methods We developed a MN model with the modified Border meth-od. At the end of the 1st, 2nd, 3th, and 4th week of immunization, respectively, the levels of whole blood uPA, tPA and PAI-1 were determined by ELISA. The rat kidney tissues were examined by light microscopy and electron microscopy to i-dentify the pathological changes. The expression levels of nephrin and WTl were detected with immumofluorescence staining and their correlation was analyzed. Results Compared the treatment group with control group, the levels of whole blood uPA, tPA and PAI-1 of the model group were decreased, while PAI-1 was elevated, showing a significant difference ( P<0. 05). The degree of renal interstitial fibrosis was more serious. Correlation analysis showed that the whole blood tPA and uPA levels were positively correlated with the changes of nephrin protein expression in the kidney tissue, while the whole blood PAI-1 level was negatively correlated with the nephrin protein expression in the kidney tissue. Conclusions In the process of MN development, the fibrinolytic system may have important significance for podocyte apoptosis. Determination of early phase of MN podocyte injury may be another therapy target for prevention of the disease development, and then pro-vide new ideas for clinical research and drug development for MN.

DNA barcoding is an effective technique in species identification. To determine the candidate sequences which can be used as DNA barcode to identify in Papaver genus, five potential sequences (ITS, matK, psbA-trnH, rbcL, trnL-trnF) were screened. 69 sequences were downloaded from Genbank, including 21 ITS sequences, 10 matK sequences, 8 psbA-trnH sequences, 14 rbcL sequences and 16 trnL-trnF sequences. Mega 6.0 was used to analysis the comparison of sequences. By the methods of calculating the distances in intraspecific and interspecific divergences, evaluating DNA barcoding gap and constructing NJ and UPMGA phylogenetic trees. The sequence trnL-trnF performed best. In conclusion, trnL-trnF can be considered as a novel DNA barcode in Papaver genus, other four sequences can be as combination barcode for identification.
DNA Barcoding, Taxonomic ; methods ; Papaver ; classification ; genetics

Objective To observe the effect of the electroacupuncture ( EA) on the expression of cyclin-de-pendent kinase 5 ( Cdk5 ) in rats with muscle contusion and to explore its mechanism. Methods Thirty-two Sprague-Dawley rats were randomly divided into a normal group of 4, a model group of 4, a natural recovery group ( NR) of 12 and an EA group of 12. All except those in the normal group had acute skeletal muscle contusion induced through a heavy blow. The EA group was treated with 15 minutes of EA daily beginning 48 h after the injury while the other rats received no EA. The model group was sacrificed 24 h after modeling, and rats from the NR and EA groups were sacrificed on the 7th, 14th and 21st day after the modeling to collect tissues. Hematoxylin eosin ( HE) staining, Western blotting and quantitative real-time fluorescence PCR were used to observe any histological changes, as well as Cdk5 protein and mRNA expression. Results The HE staining showed that the other 3 groups displayed larger a-mounts of muscle fiber fracture, dissolution and inflammatory cell invasion than was observed in the normal group. Compared with the NR group, quicker recovery was seen in the EA group as evidenced by faster muscle satellite cell proliferation and more new muscle fiber generation. The average Cdk5 protein expression in both the NR and EA groups was higher than in the normal group, and that of the EA group was significantly lower than that of the NR group. Conclusions Muscle contusion can increase Cdk5 expression in skeletal muscles, at least in rats. EA can promote the restoration of skeletal muscle function, probably by inhibiting CDK5 protein and mRNA expression.

Objective To construct an evaluation index system of ward management,which can evaluate the efficiency of ward management fairly, and make the ward management more scientific and standardized.Methods Delphi method was used in semi-structured interview of 31 experts and 74 experts were subject to questionnaire consultation, so as to establish the index system.Results The index system of ward management so built consisted of three level-1 indexes of safety and quality, teamwork and patient satisfaction, six level-2 indexes of daily monitoring, service environment, adverse events, doctor-nurse cooperation, evaluation of administrators and patient satisfaction, and 25 level-3 indexes.Practice of this system in the past two years reduced adverse events and elevated quality of care.Conclusions This system as used clinically proves its operability and objectivity.

Objective To assess the clinical and pathological feature of patients with livedo vasculitis.Methods Clinical data were collected from 47 patients with livedo vasculitis,and retrospectively analyzed.All the patients were classified into three groups according to treatment strategy:anticoagulation group,anticoagulation + corticosteroids group,and anticoagulation + sulfasalazine group.Results Clinically,livedo vasculitis usually began as petechia or ecchymosis,edema with distending pain,followed by spotty necrosis which subsequently evolved into vermiculated ulcers and,finally,atrophie blanche.Pathological features included fibrinoid degeneration of and thrombus formation in small vessel walls within the superficial or deep dermis,as well as a sparse lymphocytic infiltrate around the injured vessels.The average time to onset of action of administrated drugs was (9.14 ± 3.48),(5.62 ± 1.04) and (8.23 ± 2.68) days,and time to remission was (2.57 ± 1.41),(4.06 ± 1.51) and (5.64 ± 1.32) months,in the anticoagulation group,anticoagulation + corticosteroids group and anticoagulation + sulfasalazine group,respectively.Conclusion Anticoagulation in combination with anti-inflammatory therapy appears to have a more rapid onset of action in the treatment of livedo vasculitis with a reduced recurrence rate compared with anticoagulation therapy alone.

Objective To investigate the correlation between the expression of Ku70 and Ku80 in esophageal squamous cell carcinoma and chemotherapy sensitivity and prognosis.Methods The expression levels of Ku70 and Ku80 in esophageal squamous cell carcinoma were measured by immunohistochemistry.The relationship between the expression of Ku70 or Ku80 and clinical pathology parameters,recent curative effect of concurrent chemoradiation and prognosis of the patients were analyzed.Results The positive expression rate of Ku70 and Ku80 was 59.0％ (69/117) and 50.4％ (59/117) in esophageal squamous cell carcinoma.The expression of Ku70 and Ku80 was significandy related to recent curative effect of esophageal squamous cell carcinomas (P =0.017,0.012).The patients with complete response had lower positive expression rate of Ku70 and Ku80 [49.2％(32/65),40.0％(26/65)].But the expression had no relationship with the age,length of lesions in CT,lymph node metastasis of esophageal squamous cell carcinoma (P ＞0.05).There was no relationship between the expression of Ku70 and Ku80 neither (P =0.114).KaplanMeier analysis found Ku70 and Ku80 had worse prognosis in patients with positive expression (Ku70:P =0.004 ; Ku80:P =0.025).In Cox univariate analysis concluded the same result (Ku70:P =0.005,HR =1.971,95％ CI:1.230-3.159; Ku80:P =0.028,HR =1.659,95％ CI:1.057-2.605); but in Cox multivariate analysis,only Ku70 expression was the independent factor (P =0.033).Conclusions The upregulation of Ku70 and Ku80 is correlated to decreased concurrent chemoradiation sensitivity.In addition,the upregulation of Ku70 is correlated to worse prognosis.Ku70 and Ku80 can be a prediction index for predicting concurrent chemoradiation sensitivity in esophageal squamous cell carcinoma.

Objective To explore the effect of fluoxetine on the brain function of bulimia nervosa (BN) patients.Methods Seven female BN patients,who met criteria of the 3rd version Chinese Criteria of Mental Diseases (CCMD-3),accepted functional magnetic resonance imaging (fMRI) examinations before and after the antidepressant treatment (fluoxetine (20 mg/day)) for three months.Seven normal controls accepted the same fMRI examination only at baseline.fMRI imaging was block-design.Blocks of food or non-food stimulus containing pictures selected from International Affective Picture System (IAPS) which were shown by computer automatically.All subjects were evaluated by Hamilton anxiety scale (HAMA),Hamilton depression scale (HAMD17) and Likert Scale-likelihood evaluation to the same pictures in the fMRI imaging blocks.Results The average intensity and volume activated in BN before treatment were both significantly lower than that in the control (P＜0.05).But under stimulus of food pictures,bilateral prefrontal cortex and left amygdala of BN patients were significantly activated.After fluoxetine treatment,the intensity and volume activated both increased significantly (P＜0.01) and the main areas being activated were right temporal,cerebellum and bilateral prefrontal cortex.Conclusion Fluoxetine improves the bulimic symptoms of BN patients and decreases abnormal activation of prefrontal and limbic in these areas.The underline mechanism may be related to functions of serotonin system in prefrontal-limbic path.

Objective To set up and verify a new scoring system of clock drawing test(CDT).Methods CDT and other neuropsychological tests were applied to 180 normal individuals.170 subjects with mild cognitive impairment(MCI),and 31 patients with mild Alzheimer's disease(AD).The first step of the 30-score system of CDT was to anchor the 4 points of"12-3-6-9".totaling a score of 4 which was the "30-score system A(Anchoring)part";and the other 13 items reflecting the drawing results and totaling a score of 26 made up the so called"30-score system C(Clockfaee)part".the two together had a total score of 30.Thirty-eight subjects underwent Xe-enhanced computed tomography(Xe-CT)scanning for quantitively measuring regional cerebral blood flow(rCBF)of 26 regions of brain.Results Correlations coefficients of the scores of 14 items with the total score in 30-score system of CDT were in the range of 0.48to 0.71(P＜0.01).30-score system C part was closely correlated with indicators of visuospatial and executive function,while 30-score system A part with memory indicators.The sensitivity and specificity of the 30-score system A part score≤2 for detecting MCl were 70.6%and 73.9%.respectively.The sensitivity the specificity of 30-score system C part score≤17 for mild AD were 75.3%and 75.9%.respectively.In the regression equation of 30-score system A and C part.the combined independent variables were the CBF of left frontal cortex and right white matter,respectively.Conclusion The 30-score system A and C part have difierent meanings:30-score system C part could be used to identify AD while 30-score system A part is helpful for identiring MCI.