Objective To establish a method for the preparation of long-circulating liposomes (LCL) containing etoposide and to observe its stability in rats plasma.Methods The etoposide-containing liposome was prepared by ethanol injection method. Polyethylene glycol-distearoyl phosphatidylethanolamine (PEG2000-DSPE) was used to modify the membrane of the liposome. Reversed-phase high pressure liquid chromatography (RP-HPLC) was used to detect the concentration of the liposome,and dynamic release method was used to study its stability in mice plasma.Results The mean size of the LCL containing etoposide was (180?26) nm,and the mean concentration of etoposide was (4.78?0.22) mg/mL,with the entrapment efficiency being (88.71?8.2)%. The leakage ratio of the conventional liposome containing etoposide and LCL containing etoposide in mice plasma were (80.14?1.59)% and (46.72?2.61)%,respectively.Conclusion LCL containing etoposide with high entrapment efficiency and low leakage rate can be obtained by using ethanol injection method. Additionally,modification by PEG2000-DSPE could raise the stability of the liposome.

Objective To observe the abnormity of heart function in rats with pressure overload-induced left ventricular hypertrophy and the changes of NCX,SERCA2a expression in myocardial tissues. Methods Cardiac hypertrophy was induced by clipping the abdominal aorta in rats. The cardiac hypertrophy was evaluated by Left ventricular weight index(LVWI,left ventricular weight/body weight). NCX, SERCA2a mRNA and protein expressions in left ventricular tissues were determined by half-quantitative RT-PCR and Western blot normalized to abundance of GAPDH mRNA and protein,respectively. Results LVSP and LVEDP were obviously enhanced(P

Duck circovirus(DuCV),a potential immunosuppressive virus,was investigated in Southern China from March 2006 to December 2009 by using a polymerase chain reaction(PCR)based method. In this study,a total of 138 sick or dead duck samples from 18 different farms were examined with an average DuCV infection rate of～35%. It was found that ducks between the ages of 40～60 days were more susceptible to DuCV. There was no evidence showing that the DuCV virus was capable of vertical transmission. Farms with positive PCR results exhibited no regularly apparent clinical abnormalities such as feathering disorders,growth retardation or lower-than-average weight. The complete genomes of 9. strains from Fujian Province and 1 from Zhejiang Province were sequenced and analyzed. The 10 DuCV genomes,compared with others genomes downloaded from GenBank,ranged in size from 1988 to 1996 base pairs,with sequence identities ranging from 83.2% to 99.8%. Phylogenetic analysis based on genome sequences demonstrated that DuCVs can be divided into two distinct genetic genotypes,Group I(the Euro-USA lineage)and Group II(the Taiwan lineage),with approximately 10.0% genetic difference between the two types. Molecular epidemiological data suggest there is no obvious difference among DuCV strains isolated from different geographic locations or different species,including Duck,Muscovy duck,Mule duck,Cheery duck,Mulard duck and Pekin duck.

Objective To investigate the immunological properties of Rv1009 domain. Methods BALB/c mice were immunized with Rv1009 domain three times at 2-week interval. ELISA was used to detect the antiRv1009 domain antibody titer in the sera of immunized mice sera. The spleen lymphocytes of the immunized mice were separated and the stimulation index (SI) was measured by MTT colorimetry. Levels of secreted IFN-γ, IL-10 and IL-12 upon specific antigen stimulation were detected by ELISA. The BALB/c mice immunized with Rv1009 domain were intravenously infected with MTB H37Rv. Four weeks after the final injection, the number of CFU in spleens was determined. Results The titer of the specific antibody in sera of the immunized BALB/c mice was 1:12 800. The SI of Rv1009 domain immunized group (2. 40±0. 18) was significantly higher than that of saline immunized group (0.90±0.21). The IFN-γ,IL-10 and IL-12 levels in culture supematant of spleen lymphecytes from the fusion proteins immunized mice was (1 432±30) ng/L, (503±11) ng/L and (311±11) ng/L respectively, significant different from that of saline immunized group[(256±20) ng/L, (76±6) ng/L and(56±8) ng/L,P<0.01]. Four weeks after the final injection,compared with normal saline immunized mice (6.64±0.13), dramatic reduction in MTB replication was observed in the spleen (4.86±0.14) from BALB/c mice immunized with fusion proteins following a subsequent MTB H37Rv challenge, but the protection efficacy of mice immunized with Rv1009 domain was not as good as that of BCG vaccination group (3.81±0.16). Conclusion Rv1009 domain can be used as a candidate for the new TB vaccine.

OBJECTIVETo investigate the value of detection of hepatitis C virus core antigen (HCV-cAg) for screening blood donor by using the internal reagent enzyme immunoassay (EIA) and anti-HCV antibody.

METHODSThe first and repeat assays were performed for detection of serum anti-HCV and HCV-cAg ELISA in 3972 donor's serum specimens from August to October of 2004. Twenty-five donors positive for anti-HCV were tested with HCV-cAg EIA kits and the results were compared with the results of HCV RNA determination with RT-PCR method.

RESULTSIn 3972 donor's serum samples, only 1 serum specimen was positive for HCV RNA identification among 10 specimens which were positive for anti-HCV in first assays, and only 1 serum specimens was positive for HCV RNA identification among 12 specimens positive for anti-HCV in repeat assays, only 2 serum specimens were positive HCV RNA identification in 3 specimens which were positive for HCV-cAg assays.

CONCLUSIONThe sensitivity of HCV-cAg ELISA is similar to HCV RT-PCR, but it is much cheaper. Therefore, HCV-cAg ELISA and anti-HCV may be used together to screen blood donor.

Blood Donors ; Enzyme-Linked Immunosorbent Assay ; Hepatitis C Antibodies ; blood ; Hepatitis C Antigens ; blood ; Humans ; RNA, Viral ; blood ; Reverse Transcriptase Polymerase Chain Reaction ; Viral Core Proteins ; blood

Objective To compare the effect of right ventricular outflow tract (RVOT) and right ventricular apex (RVA) pacing on ventricular systolic synchrony using gated blood pool SPECT (GBPS).Methods A total of 50 patients implanted with pacemaker due to high degree or complete atria-ventricular block were enrolled in the study. Twenty-three patients were RVOT paced ( Group A, n = 23) and 27 were RVA paced (Group B, n=27). Twenty-four patients with malignancy, normal echocardiographic findings and no history of cardiac diseases were scheduled for pre-chemotherapy evaluation of cardiac structure and function and were enrolled as control group ( Group C, n = 24). All patients underwent GBPS imaging and the values of phase angle (PS), mean phase of each wall, standard deviation (SD) of mean phase of each wall, lateral-septal motion delay of left ventricle ( LV Sep-Lat Delay), septal-right ventricular (RV) delay of LV ( LV Sep-RV Delay) and LV-RV Delay were acquired. The parameters of ventricular systolic synchrony among the three groups were compared using one-way ANOVA. Results The mean phase of LV lateral wall in Groups A and B were significantly higher than that in Group C: Group A (120.50 ±40.58) ms; Group B (103.23±28.34) ms; Group C (84.63 ±22.38) ms (F=7.72, P ＜0.05). There was no significant difference between Groups A and B ( t = 1.30, P ＞ 0.05 ). The mean phase of RV in Group A was significantly larger than those in Groups B and C: Group A ( 137.05 ± 39.27) ms, Group B ( 100.85 ± 23.79) ms,Group C (59. 13 ±30.52) ms (F=35.55, P＜0.05). PS, SD and LV Sep-Lat Delay in Groups A and B were significantly higher than those in Group C: (85.73 ± 12.00)°vs (89.85 ± 15.61 )°vs (58.95 ±9.87)°, (27.68±10.66) ms vs (26.15 ±13.02) ms vs (15.63 ±8.35) ms, (25.06±34.23) ms vs (2. 62 ± 60. 31 ) ms vs ( - 23.66 ± 31.39) ms, F = 41.54,8.55,6.81, all P ＜ 0.01 ), however, there was no significant difference between Groups A and B ( t = 0. 68, 0.68, 1.30, all P ＞ 0.05 ). LV Sep-RV Delay and LV-RV Delay were significantly different among the three groups ( LV Sep-RV Delay: Group A (57.60 ±56.77) ms, Group B (6.36 ±61.88) ms, Group C ( -41.89 ±35.78) ms; LV-RV Delay:Group A (47.36 ±42.59) ms, Group B ( 3.08 ± 38.81 ) ms Group C ( - 26.50 ± 20.99 ) ms, F = 20. 32,25.38, both P ＜ 0.01 ). Conclusion Both RVA and RVOT pacing increase the segmental phases detected by GBPS, causing inter- and intra- ventricular asynchrony compared with patients without pacemakers.

OBJECTIVETo evaluate the erectile and ejaculatory function of sacral tumor patients after sacral nerve root resection and investigate the relationship of erectile and ejaculatory dysfunction (EED) with the level of sacral nerve injury.

METHODSThis retrospective study included 47 male patients aged 16 to 63 (32.6 +/- 6.8) years treated by sacral tumor resection between January 2008 and August 2013. According to the levels of the sacral nerve roots spared in surgery, the patients were divided into four groups: bilateral S1-S3 (n=16), unilateral S1-S3 (n=21), unilateral S1-S2 (n=6), and unilateral S1 (n=4). The patients were followed up for 12 to 41 (27.2 +/- 10.9) months by questionnaire investigation, clinic review, and telephone calls about their erectile and ejaculatory function at 3, 6 and 12 months after surgery and in August 2013.

RESULTSIn the bilateral S1-S3 group, the incidence rates of EED were 31.25% (5/16), 25% (4/16), and 12.5% (2/16) at 3, 6, and 12 months respectively after surgery, with recovery of erectile and ejaculatory function in August 2013. The incidence rates of EED in the unilateral S1-S3 group were 85.71% (18/21), 71.43% (15/21), 52.38% (11/21), and 42.86% (9/21) at 3, 6 and 12 months and in August 2013, respectively; those in the unilateral S1-S2 group were 100% (6/6), 83.33% (5/6), 83.33% (5/6), and 66.67% (4/6) at the four time points; and those in the unilateral S1 group were all 100% (4/4). No statistically significant differences were found in the incidence rate of EED among the patients of different ages or tumor types (P > 0.05).

CONCLUSIONThe incidence of postoperative EED in male patients treated by sacral tumor resection is closely related to the mode of operation. Sparing the S3 nerve root at least unilaterally in sacral tumor resection is essential for protecting the erectile and ejaculatory function of the patient.

Adolescent ; Adult ; Ejaculation ; physiology ; Erectile Dysfunction ; epidemiology ; etiology ; Female ; Humans ; Incidence ; Male ; Middle Aged ; Organ Sparing Treatments ; Peripheral Nervous System Neoplasms ; surgery ; Postoperative Complications ; epidemiology ; Postoperative Period ; Retrospective Studies ; Sacrum ; Spinal Nerve Roots ; injuries ; surgery ; Surveys and Questionnaires ; Young Adult

Objective To study the phenomena and the related mechanisms of malignant transformation of dermis derived multipotential stem cells in vitro . Methods Clonal populations of dermal multipotential stem cells were passaged sequentially in vitro , and the subcutaneous inoculation of cells in nude mice was used for observation of the tumor formation. The transcript profiles of the transformed cells were analyzed by DNA microarray technique. Results Dermal multipotential stem cells underwent spontaneous malignant transformation after serial subculture in vitro . Cells grew out of control, and chromosome number was abnormal. After cells were inoculated subcutaneously into BALB/c nu/nu athymic mice, tumors characterized by fibrous histiocytoma were produced. Immunohistochemistry showed that there were different cell populations for the expression of vimentin, cytokeratin, S 100, and ? smooth actin. Detection by DNA microarray technique revealed that the transformed cells expressed multilineage transcripts, indicating that the transformed cells might have the multipotency. Among the differentially expressed genes in transformed cells, most of the up regulated genes were related to the proliferation process, but most of the down regulated genes were growth factors and their receptors. The enhanced expression of the c ki ras gene and its relevant molecules may play important roles in the transformation process. A candidate gene with unknown functions related to the stem cell proliferation was also preliminarily identified. Conclusion Dermal multipotential stem cells can undergo spontaneous malignant transformation in vitro . Further studies of the mechanisms of this process at the molecular level may have significance both in stem cell application and in tumorigenesis.

OBJECTIVETo screen and evaluate the active constituents of Chinese medicinal herbs as potent inhibitors of Cdc25 phosphatase.

METHODSThe affinity chromatography purified glutashione-S-transferase/Cdc25A phosphatase fusion protein and Cdc2/cyclin B from the extracts of starfish M phase oocytes are used as the cell cycle-specific targets for screening the antimitotic constituents. We tested 9 extracts isolated from the Chinese medicinal herbs and vegetables including the agents currently used in cancer treatment by measuring the inhibition of Cdc25A phosphatase and Cdc2 kinase activity. The antitumor activity of the extracts was also evaluated by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay and flow cytometry.

RESULTSCdc25A inhibitory activity and antitumor activity are detected in the extracts isolated from three Chinese medicinal herbs Agrimona pilosa; Herba solani lyrati; Galla chinesis.

CONCLUSIONWe found three extracts isolated from Chinese medicinal herbs have potential inhibitory activity of Cdc25 phosphatase using a highly specific mechanism-based screen assay for antimitotic drug discovery.

Apoptosis ; drug effects ; Cell Cycle Proteins ; antagonists & inhibitors ; metabolism ; Cell Line, Tumor ; Cyclin-Dependent Kinases ; metabolism ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; analysis ; chemistry ; pharmacology ; Humans ; Lethal Dose 50 ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; drug therapy ; enzymology ; pathology ; Medicine, Chinese Traditional ; methods ; Mitosis ; drug effects ; Phytotherapy ; methods ; Plants, Medicinal ; chemistry ; cdc25 Phosphatases ; antagonists & inhibitors ; metabolism

OBJECTIVETo evaluate the significance of cardiac output (CO) response against exercise determined by IGR method and LVEF, 6 MWT distance in patients with chronic heart failure (CHF).

METHODTo adopt 6 MWT, and before and after the test measuring the CO by the IGR method, furthermore, measure LVEF to 36 patients (heart failure group) with CHF, compare with the health groups (control group).

RESULTSThe 6MWT distance of heart failure group (333.00 +/- 49.64) m decrease compared with the control group (582.56 +/- 67.97) m (P < 0.01), moreover, the distance of NYHA class III (314.82 +/- 36.27) m is significantly shorter than II (361.57 +/- 55.42) m (P < 0.05). The LVEF of heart failure group (47.0 +/- 0.4)% reduce compared with the control group (66.9 +/- 5.2)% (P < 0.01), and the data of NYHA class III (43.3 +/- 10.3)% is significantly lower than II (52.8 +/- 7.6)% (P < 0.01). The increase in CO response against exercise of heart failure group (5.97 +/- 1.89) L/min decrease compared with control group (8.88 +/- 0.52) L/min (P < 0.01), furthermore, the value of NYHA class III (5.31 +/- 1.52) L/min, compared with II (7.01 +/- 1.98)L/min, is obviously lower (P < 0.01). The 6MWT distance correlates positively with the increase in CO response against exercise (r = 0.63, P < 0.01), but the correlation is not found between the increase CO response against exercise and the LVEF (r = 0.11, P > 0.05).

CONCLUSIONOur results show that CO response against exercise measured by IGR method, with the advantages of being noninvasive, safe, convenient and accurate, combining with the 6MWT can evaluate cardiac reserve in patient with CHF.

Adult ; Aged ; Cardiac Output ; Case-Control Studies ; Chronic Disease ; Exercise Test ; Exercise Tolerance ; Female ; Heart Failure ; diagnosis ; physiopathology ; Humans ; Male ; Middle Aged ; Walking