Objective To construct human phage antibody library against hepatoma carcinoma.Methods Peripheral blood mononuclear cells(PBMCs) of patients with liver cancer were sensitized in vitro and transformed by Epstein-Barr virus(EBV).The genes of light chain and Fd of antibodies were amplified by RT-PCR.Fab genes were cloned into vector pComb3 and transformed into E.coli XLI-Blue by electroporation to construct the Fab-displaying phage antibody library.Results ELISA detection showed that 4 liver cancer patients' B cells transformed by EBV could produce specific antibodies to hepatoma carcinoma cell.Totally 13 types of light chain genes and 28 types of Fd genes were obtained by RT-PCR.The capacity of the primary phage library was 1.7?107pfu/mL.The percentage of recombinant clones was about 100%.Conclusion A human phage antibody library has been constructed successfully by means of EBV transformation technique.

Objective The present study investigated the expression of heat shock cognate protein 70 (Hsc70) in the synovial tissues and blood samples of patients with rheumatoid arthritis (RA) to determine the pathological role of this protein in the pathogenesis of the disease.Methods The expression of Hsc70 in synovial membranes was quantitatively analyzed by immunohistochemistry,real-time quantitative PCR and western blotting.The samples from osteoarthritis (OA) and ankylosing spondylitis (AS) were used as controls.The levels of Hsc70 in blood of patients with RA were determined using enzyme linked immunosorbent assay (ELISA) with the samples of the healthy subjects as controls.Statistical analysis was conducted with one-way ANOVA,LSD test and Spearmen's correlation.Results Immunohistochemistry showed that Hsc70 had significantly increased expression in synovial tissues of RA than in the samples of OA and AS.Real-time PCR and western blotting confirmed the above findings.ELISA detected significantly elevated level of Hsc70 in blood of patients with RA as compared with samples from the controls (P＜0.01).Conclusion The study suggests that the up-regulation of Hsc70 may be involved in the pathogenic process of RA.

Objective To investigate the clinical value of contrast-enhanced ultrasound(CEUS) in the diagnosis of postnatal placenta increta. Methods Twenty-six patients with postnatal placenta increta were examined by gray-scale and color Doppler ultrasound and CEUS. Then microvascular perfusion and enhanced features of lesions, myometrium and serous layer were observed. Arriving time (AT), time to peak intensity (TTP) and the lasting time of enhancement (LTE) were recorded. AT,TTP and LTE of enhanced lesions were compared with those of normal myometrium. Results Serous layer, lesions and adjcent myometrium,normal myometrium enhanced in turn. There was no obvious boundary between the lesions enhanced and adjcent myometrium. AT and TTP of the lesions enhanced were both less than those of normal myometrium ( P ＜0. 05). LTE of the part of lesions enhanced was more than that of normal myometrium ( P ＜0.05).Part of lesions never enhanced during the whole process. The serous layer of uterine was smooth and uninterrupted in 24 patients. These 24 patients all recovered after conservative treatment. The local serous layer adjcent lesions was not smooth, but no contrast agent leakage occurred in another 2 patients, and uterine lobectomy were performed in emergency because of massive hemorrhage during conservative treatment. Conclusions Microvascular perfusion and enhanced features of lesions,myometrium and serous layer could be showed clearly through CEUS.

Objectives To evaluate the effects of combined atorvastatin and probucol use on endothelial function in patients with acute coronary syndrome (ACS). Methods Thirty patients with ACS were randomized to receive atorvastatin (20 mg/d) and probucol (500 mg/d, combination group, n = 15) or atorvastatin (20 mg/d) alone (atorvastatin group) within 24 h after admission for 4 weeks. Endothelium-dependent flow-mediated dilatation (FMD) and endothelium-independent sublingual nitroglycerin-mediated dilatation (NMD) as well as the levels of lipids and C-reactive protein were assessed at baseline, 1 week and 4 weeks after therapy. Results Compared to baseline, the levels of total cholesterol, LDL-C and C-reactive protein were significantly reduced after 1 week and 4 weeks in both groups, FMD equally increased after 1 week in both groups (atorvastatin group: 3.75%±0.78% vs. 1.09%±0.44%, combination group: 3.67%±0.36% vs. 1.24%±0.37%, P<0.01). Post4 weeks therapy, FMD increase was significantly higher in combination group (3.67%±0.36% at 1 week vs. 6.85%±0.64% at 4 weeks, P <0.01) than that in atorvastatin group (3.75%±0.78% vs. 3.80%±0.31%, P=0.954). NMD also equally and increased over 4 weeks in two groups (P<0.01 vs. baseline). There was no correlation between the change in FMD/NMD and the changes in lipids or C-reactive protein levels. Conclusions The combined atorvastatin and probucol therapy early after ACS is superior to atorvastatin alone on improving endothelial function.

Objective To observe the effects of soluble epoxide hydrolase inhibitor t-AUCB on foam cell formation and cholesterol efflux in macrophage.Methods Mouse macrophages RAW264.7 were cultured and stimulated with ox-LDL (80 μmol/L) in the absence (group A) or presence of t-AUCB (1,10,50,100 μmol/L,group B) or t-AUCB (100 μmol/L) pretreated with PPARγ antagonist GW9662 (5 μmol/L,group C).The foam cell was identified by oil red O staining.The cholesterol efflux rates of 3 Hcholesterol in cells were measured by liquid scintillation counter.mRNA and protein expressions of ABCA1 were detected by real-time PCR or Western blot,respectively.Results Oil red O staining showed that t-AUCB (100 μmol/L) significantly inhibited foam cell formation which could be significantly reversed by GW9662 (all P ＜ 0.05).t-AUCB dese-dependently increased cholesterol efflux rates in mouse macrophage [(5.91+0.18)％ in group A,(7.03 ±0.33)％,(8.05 ±0.32)％,( 9.04 ±0.14)％,(10.06±0.85)％ in 1,10,50,100 μmol/L t-AUCB groups,all P＜0.05 vs.group A],which could be reversed by pretreatment with GW9662 [ (6.33 ±0.15)％ in 100 μmol/L t-AUCB + GW9662 group].t-AUCB also upregulated ABCA1 mRNA and protein expressions in a dose-dependent manner which could be significantly attenuated by pretreatment with GW9662.Condusion t-AUCB could inhibit foam cell formation by improving cholesterol efflux through activating PPARγ-ABCA1 pathway in macrophage.

Objective To further evaluate the safety and observe for unknown adverse reactions in the prelicensure trials of Anflu(split influenza virus vaccine)in children aged 6 months to 3 years old and 3 to 11 years old respectively.Methods This open clinical trial enrolled 100 healthy children in each of the two groups:6 months to 3 years old group and 3 to 11 years old group.The 6 months to 3 years group were vaccinated with two pediatric doses(0.25 ml/dose),28 days apart.The 3 to 11 years group were vaccinated with one adult dose(0.5 ml/dose).All the subjects were observed for 30 min after vaccination and had 3 follow-up visits at 24,48,72 h after vaccination. All subjects with adverse reactions were followed up till the symptoms resolved.Results The total adverse reaction rate was 6.0%(12/200).The occurrence rates of local reaction and systemic reaction were 1.0%(2/200)and 5.5%(10/200)respectively.For the younger group and older group,the adverse reaction rates were 8.0%and 4.0%respectively.Conclusion This vaccineis safe in children aged 6 months to 11 years old.

Objective To further evaluate the safety and observe for unknown adverse reactions in the prelicensure trials of Anflu(split influenza virus vaccine)in children aged 6 months to 3 years old and 3 to 11 years old respectively.Methods This open clinical trial enrolled 100 healthy children in each of the two groups:6 months to 3 years old group and 3 to 11 years old group.The 6 months to 3 years group were vaccinated with two pediatric doses(0.25 ml/dose),28 days apart.The 3 to 11 years group were vaccinated with one adult dose(0.5 ml/dose).All the subjects were observed for 30 min after vaccination and had 3 follow-up visits at 24,48,72 h after vaccination. All subjects with adverse reactions were followed up till the symptoms resolved.Results The total adverse reaction rate was 6.0%(12/200).The occurrence rates of local reaction and systemic reaction were 1.0%(2/200)and 5.5%(10/200)respectively.For the younger group and older group,the adverse reaction rates were 8.0%and 4.0%respectively.Conclusion This vaccineis safe in children aged 6 months to 11 years old.

OBJECTIVETo measure the iodine nutritional status on adult islanders and to evaluate the advantages and disadvantages of iodized salt prophylactic programs.

METHODSA comparative study was carried out in 8 rural townships selected from Dinghai (iodized salt district) and Daishan (non-iodized salt district) of Zhoushan islands by random sampling method. Mann-Whitney test was used to compare the urinary iodine concentration and dietary iodine intake of the two groups. Spearman correlation test was used to look for the correlation of urinary iodine concentration and dietary iodine intake in the two groups respectively.

RESULTSThe amounts of daily iodine intake excluding the iodine intake from iodized salt in the two groups were 128 micro g and 147 micro g respectively but the difference was not statistically significant (u = 1.847, P = 0.065). The urinary iodine concentration of non-iodized salt group was 90 micro g/L, lower than 194 micro g/L in iodized salt group (u = 14.673, P = 0.000). There was no significant correlation between daily iodine intake and urinary iodine concentration (r(s) = 0.052, P = 0.095).

CONCLUSIONSIn Zhoushan islands, the daily iodine intake did not meet the daily need (150 micro g/day) suggesting that iodized salt supplement was necessary. However, side effect due to overdose should be brought into attention.

Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Goiter ; epidemiology ; Humans ; Infant ; Iodine ; administration & dosage ; adverse effects ; deficiency ; urine ; Male ; Middle Aged ; Sodium Chloride, Dietary ; administration & dosage ; adverse effects

OBJECTIVECord blood (CB) provides a rich source of stem cells for transplantation. CB transplantation has been used widely after myeloablative therapy. One major disadvantage of CB transplantation is delayed platelet engraftment. The aim of this study was to hasten platelet engraftment by investigating the ability of different hematopoietic growth factor combinations to generate large numbers of megakaryocyte (Mk) from CB and by evaluating the biologic characteristics and function of the expanded Mk.

METHODSCB samples were obtained at the end of normal full-term deliveries with informed consent. Mononuclear cells (MNCs) were isolated from CB using Ficoll density centrifugation. MNC population was positively selected for CD(34) expression by magnetic cell sorting (MACS). CD(34)(+) cells were cultured in serum-free and stroma-free medium containing the following two different cytokine combinations: thrombopoietin (TPO) + stem cell factor (SCF) + interleukin (IL) -3 + IL-6 and TPO + SCF. Cultures were characterized after 3, 7, 10 and 14 days by flow cytometry, colony forming unit-megakaryocyte (CFU-Mk) and maturation evaluation (Mk ploidy). The expanded Mk function was examined by the platelet activation in vitro and severe combined immunodiffiency (SCID) mice transplantation in vivo.

RESULTSDifferent results were observed with different culture conditions. With the first cytokine combination optimal expansion of CD(41)(+) cells was observed on day 10, but the optimal expansion of Mk progenitors (CD(34)(+)CD(41)(+)) was observed on day 7, with a median 121 and 44-fold increase at the starting cell dose. This result was also proven by CFU-Mk. The largest numbers of CFU-Mk were also observed on day 7. The degree of maturation of Mk cells also increased as suggested by DNA content of CD(41)(+) cells, which means that CD(34)(+) cells cultured for 3 - 7 days were richer in primitive Mks, while those cultured for 10 - 14 days had greater numbers of more differentiated Mks. For the second cytokine combination, CD(41)(+) and CD(34)(+)CD(41)(+) cells were fewer than the first one, but it produced 36 and 85-fold CD(34)(+)CD(41)(+) and CD(41)(+) respectively on day 7. Platelet activation test confirmed that the expanded Mks had normal function. Therefore, the expanded Mks could be transplanted into the SCID mice bone marrow and produce human platelet in the peripheral blood of the mice.

CONCLUSIONEx vivo expanded Mk might facilitate CB transplantation and help shorten the period of post-transplant thrombocytopenia.

Animals ; Antigens, CD34 ; Cell Culture Techniques ; methods ; Cells, Cultured ; Culture Media ; Fetal Blood ; cytology ; Humans ; Leukocytes, Mononuclear ; cytology ; Megakaryocytes ; cytology ; Mice ; Mice, SCID

OBJECTIVETo explore the impact of microwave radiation on GC-2spd cells.

METHODSWe exposed cultured GC-2spd cells to microwave radiation at the average power densities of 0, 10 and 30 mW/cm2 for 15 minutes and, from I to 24 hours after the exposure, we observed the changes in cell proliferation, histology and ultrastructure, cell apoptosis, and cAMP content by MTIT, light microscopy, electron microscopy, flow cytometry and ELISA.

RESULTSCompared with the control group, the GC-2spd cells showed a significant decrease in proliferation ability at 1 -24 hours after 10 and 30 mW/cm2 microwave radiation, except at 12 hours after 30 mW/cm2 radiation (P <0.05 or P <0.01), with reduced length and number of cell enation and increased intra cytoplasm vacuoles. The rate of cell apoptosis (%) was significantly increased in the 10 and 30 mW/cm2 groups at 6 hours (4.56 +/- 2.09 vs 14.59 +/- 1.09 and 8.48 +/- 1.73, P <0.05 or P <0.01) , with agglutination and margin translocation of chromatins and obvious dilation of endo cytoplasmic reticula. The cAMP content (nmol/g) in the GC-2spd cells was remarkably reduced in the 10 and 30 mW/cm2 groups at 6 and 24 hours (2.77 +/-0.24 vs 1.65+/- 0. 17 and 1.96+/-0.10, 3.02 +/-0.47 vs 2.13 +/-0.33 and 1.69 +/-0.27, P <0.05 or P <0.01).

CONCLUSIONMicrowave radiation at 10 and 30 mW/cm2 may cause injury to GC-2spd cells, which is manifested by decreased content of intracellular cAMP, reduced activity of cell proliferation, and increased rate of cell apoptosis.

Animals ; Apoptosis ; radiation effects ; Cell Line ; radiation effects ; Cell Proliferation ; radiation effects ; Male ; Mice ; Microwaves ; adverse effects ; Spermatocytes ; radiation effects