Objective To investigate the relationship of the classification of Doppler ultrasound blood flow signals with matrix metalloprotei-nase-9 (MMP-9)expression and microvessel density (MVD)in primary hepatocellular carcinoma (PHC).Methods Sixty patients with PHC,who underwent surgical resection in the Second Affiliated Hospital of Dalian Medical University from April 2008 to April 2013, were included in the study.These patients underwent ultrasound examination before operation.The classification of Doppler ultrasound blood flow signals in the focus was recorded.The expression of MMP-9 and CD3 1 in carcinoma tissues was detected by immunohistochemistry, and MVD was calculated.Continuous data were analyzed by t test,while categorical data by chi-square test;the relationship was evaluated by Spearman correlation analysis.Results The classification of Doppler ultrasound blood flow signals was as follows:grade 0 (15 cases);grade 1 (20 cases);grade 2 (15 cases);grade 3 (10 cases).The positive rate of MMP-9 expression was 72.3%.The MVD was 43.2 ± 5.4.The classification of Doppler ultrasound blood flow signals in the focus was positively correlated with MMP-9 expression and MVD (r=0.56,P<0.05).Conclusion The classification of Doppler ultrasound blood flow signals can reveal vascular changes in the focus, and it can be used in the clinical evaluation of angiogenesis in tumor tissues and the clinical diagnosis and treatment of PHC.

Objective To establish a stable method for acute isolation of atrial myocytes in diabetic rats,so as to provide materials for electrophysio?logical study of diabetic rats. Methods Streptozotocin(STZ)was used to establish type I diabetic model. Atrial myocytes were isolated with modi?fied perfusion buffer by Langendorff perfusion. The atrial myocytes were morphologically observed with optical microscope and identified by morphol?ogy and immunofluorescence staining. The action potential was recorded by patch clamp technique. Results STZ can establish a stable type I dia?betic model. The modified perfusion method can yield calcium tolerant and spindle shaped atrial myocytes. Immunofluorescence indicated that atrial myocytes were positive for Kv1.5 which was expressed in atrial myocytes. The atrial myocytes obtained by this method were able to generate action po?tentials. Conclusion The modified perfusion method is suitable for acute isolation of atrial myocytes in rats with diabetes mellitus,which may help to study the electrophysiology of diabetic heart.

ObjectiveBased on ultra performance liquid chromatography-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF-MS^E） technique， we identified qualitatively the metabolites of aristolochic acid（AAs） in rat in order to analyze the metabolic differences between water extract of Aristolochiae fructus（AFE） and Aristolochic acid Ⅰ（AAⅠ）. MethodSD rats were selected and administered AFE（110 g·kg^-1·d^-1） or AAⅠ（5 mg·kg^-1·d^-1） by oral for 5 days， respectively. Serum， urine and feces were collected after administration. Through sample pretreatment， ACQUITY UPLC BEH C₁₈ column（2.1 mm×100 mm， 1.7 μm） was used with the mobile phase of 0.01% formic acid methanol（A）-0.01% formic acid water（B， containing 5 mmol·L^-1 ammonium acetate） for gradient elution（0-1 min， 10%B； 1-7 min， 10%-75%B； 7-7.2 min， 75%-95%B； 7.2-10.2 min， 95%B； 10.2-10.3 min， 95%-10%B； 10.3-12 min， 10%B） at a flow rate of 0.3 mL·min^-1. Positive ion mode of electrospray ionization（ESI⁺） was performed in the scanning range of m/z 100-1 200. In combination with UNIFI 1.9.4.053 system， the Pathway-MS^E was used to qualitatively analyze and identify the AAs prototype and related metabolites in biological samples（serum， urine and feces）， and to compare the similarities and differences of metabolites in rats in the subacute toxicity test between AFE group and AAⅠ group. ResultCompared with AAⅠ group， 6， 10， 13 common metabolites and 14， 20， 30 unique metabolites were identified in biological samples（serum， urine and feces） of AFE group， respectively. Moreover， the main AAs components always followed the metabolic processes of demethylation， nitrate reduction and conjugation. Compared with common metabolites in AAⅠ group， prototype components of AAⅠ in serum and most metabolic derivatives of AAⅠ［AAⅠa， aristolochic lactam Ⅰ（ALⅠ）a， 7-OHALⅠ and its conjugated derivatives］ in biological samples were significantly increased in AFE group（P<0.05， P<0.01）， except that the metabolic amount of ALⅠ in feces of AFE group was remarkably lowed than that of AAⅠ group（P<0.01）. In addition， a variety of special ALⅠ efflux derivatives were also identified in the urine and feces of the AFE group. ConclusionAlthough major AAs components in AFE all show similar metabolic rules as AAⅠ components in vivo， the coexistence of multiple AAs components in Aristolochiae Fructus may affect the metabolism of AAⅠ， and achieve the attenuating effect by increasing the metabolic effection of AAⅠ and ALⅠ.