The research and development of innovative drugs targeting the clinical cure of chronic hepatitis B has become active in recent years. In the clinical trials of new drugs for hepatitis B, the use of new design concepts, new methods, and new technologies to evaluate the efficacy of innovative drugs is expected to shorten the clinical research process of candidate new drugs and reduce the cost of new drug development. However, the new designs such as seamless adaptive design and master plan design have few practical applications in clinical trials of innovative hepatitis B drugs. This article will focus on the methodological factors in the design of phase II/III clinical trials of innovative hepatitis B drugs, and introduce the key points of the new clinical trial design in order to provide methodological references for the development of innovative hepatitis B drugs.

Objective To explore the effects of the motivational interview on medication adherence and control level of asthma in adult patients with asthma. Methods A total of 100 asthma patients who were hospitalized in the Respiration Department of Putuo People's Hospital in 2016 were selected in the study using convenience sampling. According to the random number table, the participants were divided into the observation group (n=50) and the control group (n=50). Two cases from the observation group withdrew from the study due to personal reasons, leaving 48 patients in the observation group. Three cases from the control group withdrew from the study (i.e. two for disease changes and one for personal reasons), leaving 47 cases in the control group. The control group received conventional health education while the observation group received education based on motivational interview. The asthma medication compliance scale (Chinese version) (MARS-A) and asthma control test (ACT) questionnaire were used to evaluate the medication compliance and asthma control level before the intervention and 1, 2 and 3 months after the intervention. Results None of the patients had good medication compliance before the intervention. However, patients in the observation group had significantly better adherence at 1, 2, and 3 months after the intervention compared with the control group (P<0.05). There were no significant differences in asthma control level between two groups before intervention (P>0.05). But after the intervention, the repeated variance analysis showed that there were significant differences in both intervention effect and time effect for asthma control score, and the interaction effects existed between intervention and time (P< 0.05). Conclusions The health education based on motivational interview can effectively improve the medication adherence and the control level of asthma in adult patients with asthma.

Objective:To research the imumunological effect of dendritic cell transfected by recombinant plasmid of ΔNp73αon breast cancer.Methods:Cultivate dendritic cells by cytokines( GM-CSF,IL-4,TNF-α) from human umbilical cord blood in vitro, CD1a,CD83 were detected by flow cytometry.Recombinant plasmid pcDNA-HA/ΔNp73αwas used to transfected DCs,the transfection was detected by Western blot.Transfected DCs cocultivate with T cell induce specific cytotoxic lymphocytes( CTL).Proliferation ability of T cell was detected by MTT methods.IFN-γin the culture supernatant of the transfected DCs were detected by ELISA.The killing effect on breast cancer MDA-MB-231 was detected by LDH method.Results:The expression of CD1a(56%),CD83(74%)in mature DC was higher than immature DC(CD1a 19%,CD83 13%)(P<0.01).ΔNp73αtransfected group had a special band.The killing effect of specific CTL induced byΔNp73α-DC and T cells cocultivation on breast cancer cell MDA-MB-231 was statistic stronger than DC group(P<0.05),the proliferation of T cell and secretion level of IFN-γcompared with DC and pcDNA group were statistically significant(P<0.01).Conclusion:The CTL induced byΔNp73αtransfected DC vaccine has significant effect on killing breast cancer.

Objective To explore the feasibility of contrast-enhanced ultrasonograhy (CEUS) in preoperative classification of hepatocellular carcinoma (HCC) microvascular morphology.Methods Totally 94 HCC patients who underwent CEUS were analyzed retrospectively. And the offline Time-intensity curve (TIC) were drawn using SonoTumor. The tumor size, alpha fetoprotein (AFP), cirrhosis, Child-Pugh classifi cation, tumor differentiation and TNM stage were statistically analyzed. The intratumoralmicrovessels of HCC in 94 cases were evaluated by CD34 immunohistochemical staining. The relationship between intratumoral microvessel morphology and CEUS parameters were analyzed.Results CD34 immunohistochemical staining showed three distinct microvessel types in 94 cases of HCC: 28 cases of capillary-like type, 14 cases of sinusoid-like type and 52 cases of mixed type. There were no significant differences of clinical data among three microvascular morphology types. The parameters of peak strength (PE), rise time (RT), wash-in rate (WiR), wash-in perfusion index (WiPI), wash-in area under the curve (WiAUC) and mean transit time (MTT) in 28 cases of capillary-like type were (4350.7±2566.0) a.u, (10.7±3.2) s, (717.0±489.9) a.u, (12820.3±8331.6 )a.u, (128 240.8±74 487.1) a.u, (71.9±33.1)s. Those parameters in 14 cases of sinusoid-like type were (2471.6±1107.1) a.u, (16.2±4.2)s, (321.9±171) a.u, (5 561.4±2 938.0) a.u, (86 780.1±47 563.7) a.u, (117.8±69.6)s. And in 52 cases of mixed type they were (3563.2±2343.1) a.u, (14.1±4.8)s, (519.4±403.2) a.u, (9 015.3±6 884.7)a.u, (128 240.8±74 487.1) a.u, (71.9±33.1) s respectively. The CEUS parameters of WiR, WiPI in capillary-like type HCC patients were higher than sinusoid-like type and mixed type HCC patients, while RT was lower than sinusoid-like type and mixed type HCC patients, and the differences were signifi cant (WiR: t=3.87, 3.3, bothP=0.05; WiPI: t=2.96, 2.06, bothP=0.05; RT: t=3.19, 2.34, bothP=0.05). The parameter of PE in capillary-like type HCC patients were signifi cantly higher than that in sinusoid-like type HCC patients (t=2.51,P=0.05). And the parameter of PE in capillary-like type HCC patients was higher than mixed HCC patients, but there was no signifi cant difference. The parameters of PE, WiR and WiPI in mixed type HCC patients were higher than that in sinusoid-like type HCC patients, while RT in mixed type HCC patients were lower than that in sinusoid-like type HCC patients, but there were no signifi cant differences. No signifi cant differences of WiAUC and MTT were observed in HCC patients with different microvascular morphology.Conclusions There were signifi cant differences of CEUS parameters in different microvascular morphology types. And CEUS, as a non-invasive method, can be used for preliminary preoperative prediction of microvascular morphology in HCC patients.

A quantitative analysis method of multi-components with a single marker (QAMS) for simultaneous determination of six marker compounds (one from phenolic acids and five from phthalides) in Chuanxiong Rhizoma was established by applying HPLC and using butylidenephthalide as the internal reference substance. And also the feasibility and accuracy of the established method for quality evaluation and application of Chuanxiong Rhizoma were investigated and validated. The analysis was performed with the mobile phase consisting of acetonitrile - 0.2% aqueous formic acid. The flow rate was 1.0 mL . min-1 and the column temperature was maintained at 30 °C. The detection wavelengths were set at 252 nm (for ferulic acid, Z-ligustilide, and butylidenephthalide) and 266 nm (for senkyunolide I, senkyunolide A, and coniferyl ferulate), separately, and 20 µL was injected for analysis with gradient elution. The results showed that there were no significant differences observed between the HPLC-QAMS method and the external standard method (RSD <5%). The relative correction factors were credible (RSD < 5%) in changed chromatographic conditions. The established HPLC-QAMS method can be accurately used for simultaneously evaluating and controlling the quality of Chuanxiong Rhizoma with multi-components.

Objective To explore the effects of BSA on hypoxia inducible factor/hypoxia response element (HIF/HRE) transcription activity in rat tubular epithelial cells (NRK-52E) with HRE-Luc reporter plasmid.Methods Luciferin activity of NRK-52E cells incubated by a medium contained BSA in varying concentration (0,5,10,20 mg/ml) and stimulus duration (24,48,72 h) was detected by dual luciferase detecting system based on HRE-Luc reporter plasmid and HIF-1 α expression was detected by Western blotting.Results HIF/HRE transcription activity of NRK-52E cells was increased in BSA incubation group (10 mg/ml,48 h) compared with blank control (BSA 0 mg/ml,48 h) [(2.59±0.35)vs (1.03±0.09),P=0.000].HIF-1α expression of NRK-52E cells was increased in BSA incubation group (20 mg/ml,48 h) compared with blank control (BSA 0 mg/ml,24 h) [(0.052±0.010) vs (0.014±0.003),P=0.000].Conclusion Albumin can increase HIF/HRE transcription activity of TEC.

Objective To evaluate the feature of level Ⅱaxillary lymph nodes from breast cancer detected by ultrasound. Methods Eighty-nine cases of breast cancers with level Ⅱaxillary lymph nodes diagnosed in Sun Yat-sen University Cancer Center from January 2008 to October 2013 were retrospectively studied, including 66 cases of breast cancer patients pre-operatively, 23 cases of breast cancer post-operatively, all cases were female. The number, size, long/short ratio, presence of a hilum, and Doppler features of the lymph nodes were observed. The accuracy sensitivity, speciifcity, positive predictive value (PPV), and negative predictive value (NPV) of each signiifcant parameter were analyzed. Results Ultrasound examination revealed more than axillary lymph nodes, the accuracy sensitivity, specificity, positive predictive value and negative predictive value of diagnosing lymph nodes were 71.9%, 56.8%, 89.7%, 87.9%and 61.4%respectively. The maximum diameter of levelⅡaxillary lymph node was more than or equal to 10 mm, the accuracy sensitivity, specificity, positive predictive value and negative predictive value of diagnosing lymph nodes were 67.4%, 58.0%, 79.5%, 78.4%and 59.6%respectively. If there were two ultrasonographic characteristics simultaneously, the diagnostic sensitivity (87.8%) and accuracy (78.6%) could be significantly improved, the specificity, positive predictive value and negative predictive value of diagnosing lymph nodes were 60.0%, 72.3%and 80.0%. Conclusion The number and size of lymph nodes in level Ⅱaxillary area are valuable sonographic characteristics for the diagnosis of benign and malignant lymph nodes.

objective To investigate the value of NT-proBNP in assessing the volume status in maintenance hemodialysis patients with non-dominant edema.Methods One hundred and forty-five patients were recruited.Bioimpedance measurements were performed for overhydration (OH).NT-proBNP was detected by colloidal gold method.Patients were divided into three groups by levels of OH variability (△ OH,equal to OH minus weight increase) as group H (hypervolemia,n=90); group N (normovolemia,n=36) and group L (hypovolemia,n=19).Hemoglobin,albumin,blood urea nitrogen and serum creatinine were assayed,blood pressure and body mass increase were recorded.Dry weight of patients in Group H were adjusted in 3 months,the relationship between NT-proBNP and volume change were assessed.Results (1) At baseline,overall plasma NT-proBNP levels were higher than normal range.The median NT-proBNP levels in group H and group N were [1318.50(IQR 717.00,3154.25) pg/ml] and [703.50 (IQR 873.00,450.50) pg/ml],respectively.NT-proBNP was positively correlated with △OH value (r=0.801,P ＜ 0.001).(2) After 3 months,NT-proBNP levels in group H was significantly lower than baseline.Forty-one patients reached normal volume range (group H1),49 patients were resistant hypervolemia (group H2).The median NT-proBNP levels in group H1 and group H2 were [685.00 (IQR 422.50,988.50) pg/ml] and [1569.00 (IQR 982.50,2500.50) pg/ml],△ OH in group H1 and group H2 were [(0.63±0.23)L] and [(1.75±0.71)L],respectively.NT-proBNP and △ OH value in two groups had significant difference (P ＜ 0.05).NT-proBNP was positively correlated with △ OH value (r=0.684,P ＜ 0.001).(3) The area under ROC curve for NT-proBNP was 0.818,95％CI (0.733～ 0.904),P ＜ 0.001,since the absolute value of normovolemia was defined as ≤ 1.The cut off value of plasma NT-proBNP was set at 962.50 pg/ml in MHD patients with non-dominant edema,the diagnostic specificity and sensitivity were 79.6％ and 73.2％.Conclusion NT-proBNP could be used to assess volume status in MHD patients with non dominant edema.

OBJECTIVETo develop a quick identification method for the sun-dried and sulfur-fumigated Angelicae Sinensis Radix used by Fourier transform infrared spectroscopy (FTIR) combined with second derivative infrared spectroscopy.

METHODThe alcoholic and aqueous extracts of sun-dried and sulfur-fumigated Angelicae Sinensis Radix were analyzed by using FTIR, the further analysis was used by second derivative infrared spectroscopy.

RESULTThere existed differences between their infrared spectra either extracted by ethanol or water, while the distinctions were more obvious after analyzing their alcoholic and aqueous extracts through high resolution of second derivative infrared spectroscopy. Infrared spectra showed that the absorption peaks of Angelicae Sinensis Radix were significantly reduced and a new absorption peak appeared after sulfur-fumigated process in alcoholic extracts, while both of them changed markedly in the "fingerprint region" ranging from 1 000 to 400 cm(-1) in aqueous extracts. Second derivative spectra showed that the absorption peaks of sulfur-fumigated Angelicae Sinensis Radix extracted by ethanol weakened and disappeared at about 3 578 cm(-1) and 3 541 cm(-1), while both of them differed significantly from each other ranging from 1 400 to 1 200 cm(-1) as well as 800 cm(-1) to 600 cm(-1), difference also existed between them extracted by water ranging from about 3 900 to 3 850 cm(-1) and 3 800 to 3 750 cm(-1).

CONCLUSIONThe FTIS method combined with second derivative can be utilized to distinguish sun-dried and sulfur-fumigated Angelicae Sinensis Radix efficiently, conveniently and accurately, and provide a basis for identification and quality control of Angelicae Sinensis Radix.

Objective To evaluate the accuracy of contrast enhanced ultrasonography (CEUS) in determining the depth of myometrial invasion in endometrial carcinoma in stage Ⅰ.Methods Seventy-six patients previously diagnosed of endometrial carcinoma by curettage of uterine underwent transabdominal sonography (TAS) and CEUS to assess myometrial invasion,among which 48 patients proved to endometrial carcinoma in stage Ⅰ after total abdominal hysterectomy and bilateral salpingo-oophorectomy were studied.The findings of TAS and CEUS to determine endometrial carcinoma IA (no myometrial involvement or invasion of the inner half of the myometrium) and IB( invasion of the outer half of the myometrium) were compared with pathology after abdominal hysterectomy.Results Twenty one tumours (43.75%,21/48) were enhanced earlier than or simultaneously as myometrium and cervix,among which 12 cases were IA stages,while 9 cases were IB stages (P<0.05);and 27 tumors (56.25%,27/48)were enhanced late than myometrium and cervix.There was no statistical difference between TAS and CEUS in detecting endometrial carcinoma in IA and IB(P>0.05).The sensitivity,specificity,positive predictive value,negative predictive value and accuracy of TAS and CEUS in diagnosing endometrial carcinoma in IB were 61.25% vs 69.23%,77.14% vs 85.71%,50.00% vs 64.28%,72.92% vs 88.23%,72.92% vs 81.25% respectively.Conclusions CEUS is not superior to TAS in detecting deep invasion of endometrial carcinoma in stage Ⅰ.