Objective To prepare the 99Tcm-survivin mRNA antisense peptide nucleic acid (PNA)and investigate its value as a gene imaging agent in tumor bearing mice and early diagnosis in tumor.Methods Survivin mRNA antisense PNA and mismatch PNA were synthesized.Four amino acids (Gly- (D)Ala-Gly-Gly) and Aba (4-aminobutyric acid) were linked to the 5' end of PNA.Gly- (D)Ala-Gly-Gly served as a chelating moiety for strong chelation of 99Tcm and Aba acted as a spacer to minimize the steric hindrance.PNAs were labeled with 99Tcm by the ligand-exchange method.The labeling efficiency and radiochemical purity were measured by HPLC and ITLC methods.There were five BALB/c nude mice bearing human lung carcinoma ( A549 ) in each of antisense PNA and mismatch PNA groups.Gene imaging of 99Tcm-survivin mRNA antisense and mismatch PNAs were performed at 1,2 and 4 h post the injection,respectively,and the T/NT ratio was measured by the method of ROI.The statistical comparisons of average values were performed with the two-group t-test for independent sample by SPSS 13.0.Results The product kept stable in vitro.The labeling efficiency of 99Tcm-survivin mRNA antisense PNA was (95.48 ±1.92)％ and more than 85％ after the incubation for24 h in serum.The radiochemical purity was ＞ 95％.The labeling efficiency of mismatch PNA was similar to the antisense PNA.99Tcm-survivin mRNA antisense PNA was especially uptaken by tumor lesion,and its accumulation reached the top at 4 h post the injection.T/NT ratios at 1,2,and 4 h were 2.70 ± 0.28,3.44 ± 0.35,4.21 ± 0.63,respectively.In the comparison,the T/NT ratio of 99Tcm-survivin mRNA mismatch PNA at 4 h (3.12 ±0.50) was significantly lower (t =2.918,P =0.019).Conclusions 99Tcm-survivin mRNA antisense PNA has high labeling efficiency,good stability and no need of purification.Its characteristic of especial uptake by tumor lesion provides the potential value in early diagnosis of tumor.

Objective To study maternal toxicity, embryotoxicity and teratogenecity of Chimonanthus salicifolius S. Y. Hu in SD rats.Methods A total of 64 successfully mated female SD rats were randomly divided in to 4 groups (16 per group), in which 3 experimental groups were daily treated with 3.75, 7.5 and 15.0 g/kg. bw test substance by lavage from 7th to 16th day during gestation respectively. Body weight and general conditions of the pregnant rats were recorded during the study. On the 20th day in pregnancy, the rats were anatomized and examined grossly, the fetuses were removed and counted, weight, length, visceral and skeletal changes were then examined. Results There was no significant difference in the conception rate, total weight gain during the pregnancy and the number of living, dead and resorbed fetuses between each dosage groups and the control group (P>0.05) . The number of the rib, sternum, the fifth sternum punctated and the parietal bone which were ossified defectively all showed no difference among the four groups (P>0.05) . Conclusion Chimonanthus salicifolius S. Y. Hu extract had no obvious maternal toxicity, embryotoxicity and teratogenecity in SD rats under this experiment condition.

OBJECTIVETo explore the different effects of systolic blood pressure (SBP) and low density lipoprotein on carotid plaques (LDL-C).

METHODSA total of 101 510 serving and retired workers of a company who participated in the health examination in 2006-2009, 5852 participants were selected as study subjects by stratified random sampling according to the age and sex ratio. These subjects took their health examination in 2010-2011 including the carotid ultrasound. Finally, 5361 eligible participants with complete data were included in the analysis. The detection and weighted rates of carotid plaques were calculated for four groups: normal SBP and LDL-C group (3524 subjects), normal SBP and high LDL-C group (356 subjects), elevated SBP and normal LDL-C group (1308 subjects) and elevated SBP and high LDL-C group (173 subjects). The effects of different baseline SBP and LDL-C on detection rates of the carotid artery plaques were analyzed by logistic regression.

RESULTSThe detection rate of carotid plaques in normal SBP and LDL-C group, normal SBP and high LDL-C group, elevated SBP and normal LDL-C group, elevated SBP and high LDL-C group was 33.7% (1186/3524), 41.3% (147/356), 64.8% (847/1308), 68.8% (119/173) (χ(2) = 425.75, P < 0.05) and the weighted detection rate was 36.0%, 42.0%, 64.5% and 68.3% respectively. For men, the detection rate was 44.2% (877/1985), 51.1% (97/190), 70.6% (657/930), 71.3% (82/115) (χ(2) = 194.02, P < 0.05) and the weighted detection rate was 31.2%, 36.1%, 49.8% and 50.3% respectively. For women, the detection rate was 20.1% (309/1539), 30.1% (50/166), 50.3% (190/378), 63.8% (37/58) (χ(2) = 180.17, P < 0.05) and the weighted detection rate was 30.9%, 46.3%, 70.3%, and 88.1% respectively. After adjusted for other risk factors, the OR (95%CI) value was 1.37 (1.05 - 1.78), 2.05 (1.74 - 2.43) and 2.12 (1.45 - 3.12) for normal SBP and high LDL-C group, elevated SBP and normal LDL-C group and elevated SBP and high LDL-C group respectively compared with normal SBP and LDL-C group.

CONCLUSIONElevated SBP and high LDL-C were risk factors of the carotid artery plaques. Compared with high LDL-C, elevated SBP may add a higher risk for carotid plaques.

Adult ; Aged ; Blood Pressure ; Carotid Stenosis ; blood ; epidemiology ; physiopathology ; Cholesterol, LDL ; blood ; Dyslipidemias ; Female ; Humans ; Male ; Middle Aged ; Risk Factors ; Systole

OBJECTIVETo investigate the role of mesenchymal stem cells (BMSCs) in multiple myeloma (MM) bone marrow (BM) microenrivonment and their effect on myeloma cells survival and bortezomib induced apoptosis.

METHODSBMSCs were derived from BM of untreated myeloma patients (MM-BMSCs) and healthy donors (HD-BMSCs), respectively. The phenotype, proliferation time and cytokine secretion of MM-BMSCs were detected and compared with HD-BMSCs. Then BMSCs were co-cultured with myeloma cell line NCI-H929 and bortezomib in vitro. The NCI-H929 cells proliferation and bortezomib induced cell apoptosis were investigated.

RESULTSMM-BMSCs and HD-BMSCs were isolated successfully. The phenotype of MM-BMSCs was similar to that of HD-BMSCs. Expressions of CD73, CD105, CD44 and CD29 were positive, but those of CD31, CD34, CD45 and HLA-DR (< 1%) negative. The proliferation time of MM-BMSCs was longer than that of HD-BMSCs (82 h vs 62 h, P < 0.05). Moreover, over-expressions of IL-6 and VEGF in MM-BMSCs culture supernatant were detected as compared with that in HD-BMSCs [(188.8 ± 9.4) pg/ml vs (115.0 ± 15.1) pg/ml and (1497.2 ± 39.7) pg/ml vs (1329.0 ± 21.1) pg/ml, respectively]. MM- BMSCs supported survival of the myeloma cells NCI-H929 and protected them from bortezomib induced cell apoptosis.

CONCLUSIONSMM-BMSCs is benefit for myeloma cells proliferation and against cell apoptosis induced by bortezomib. Over-expression of IL-6 and VEGF maybe play a critical role in these effects.

Apoptosis ; drug effects ; Bone Marrow Cells ; cytology ; Bortezomib ; Humans ; Mesenchymal Stromal Cells ; metabolism ; Multiple Myeloma ; metabolism

OBJECTIVETo explore the association between early menarche and physical fitness among adolescent girls in China.

METHODSResearch material was selected from the data of "2010 National Physical Fitness and Health Survey". Probability unit regression method was used to calculate the age of 10th percentile at menarche and menarche age before the 10th percentile was defined as early menarche(9.0-11.6 years old). A total of 1072 girls with early menarche were selected. Each girl with early menarche was matched with one girl from the same urban or rural locations who hadn't achieved menarche with the age difference less than 0.1 years. A total of 1072 girls without menarche were selected. Indicators of physical fitness included 50 m running, standing broad jump, 50 m×8 shuttle running and sit-ups. Differences of physical fitness between early menarche and without menarche were analyzed using t test stratified by age and urban/rural area. The multilevel models of single dependent variable and multiple dependent variables were used to analyze association between early menarche and physical fitness.

RESULTSA total of 2144 students were put in this research. Among girls (11.0-11.6 years old) in urban areas, 50 m running, standing broad jump and sit-ups was (9.78 ± 0.85) s, (153.81 ± 18.59) cm, and 27.79 ± 10.25, respectively for those with early menarche, while in girls without menarche was (10.01 ± 0.90) s, (149.71 ± 18.72) cm and 26.28 ± 10.11, respectively. There were significant differences between two groups on all above variables (t values were 4.02, 3.43 and 2.31, respectively with all P values <0.01). Among girls in rural, 50 m×8 shuttle-running and sit-ups was (125.22 ± 15.57) s and 24.96 ± 8.97 for those with early menarche, while it was (120.92 ± 13.06) s and 22.96 ± 9.83 for those without menarche. There were significant differences between two groups on both variables(t values were 3.89 and 2.77 with both P values < 0.01). In addition, 50 m×8 shuttle-running in girls (10.0-10.9 years old) with early menarche was (128.52 ± 15.74) s and it was (123.89 ± 13.50) s in girls without menarche. The difference was significant (t = 2.14, P < 0.05). The multilevel analysis showed that 50 m running, standing broad jump and sit-ups in girls with early menarche was 0.12 s, 3.14 cm and 1.11 higher than girls without menarche (Waldχ(2) values were 4.00, 6.22 and 4.07, respectively with all P values < 0.05). But 50 m×8 shuttle-running in girls with early menarche was 1.95 s less than girls without menarche (Waldχ(2) = 3.96, P < 0.05).

CONCLUSIONEarly menarche may be associated with higher speed fitness, leg power and muscle power and lower physical stamina.

Child ; China ; Female ; Humans ; Menarche ; physiology ; Models, Statistical ; Physical Fitness ; physiology ; Rural Population ; Students ; Urban Population

OBJECTIVETo analyze the association between abdominal obesity and blood pressure among 7-10 years old Chinese children.

METHODSA total of 40 495 children aged 7-10 years with complete height, weight, waist circumference and blood pressure data were chosen from the data of 2010 Chinese National Survey on Students Constitution and Health. Based on the "Reference Norms for Screening Overweight and Obesity in Chinese Children and Adolescents" developed by Working Group Obesity in China (WGOC) and the waist to height ratio reference value for children (WHtR ≤ 0.46) , the children were divided into 4 body types (normal weight, simple abdominal obesity, combined obesity and other types). High blood pressure was defined as the systolic blood pressure (SBP) and/or diastolic blood pressure (DBP) higher than the P 95 blood pressure value of 2010 national student of the same gender and age. The prevalence of simple abdominal obesity, combined obesity and high blood pressure were described. Three groups (with 2165 children in each) of children with normal weight, simple abdominal obesity, combined obesity respectively were selected from the chosen ones matched by gender, age (within ± 0.5 years old) and height (within ± 0.3 cm) at the ratio of 1: 1: 1. The value of blood pressure and prevalence of high blood pressure were described.χ(2) test, analysis of variance and multilevel models were used to analyze the relationship between obesity and blood pressure.

RESULTSA total of 40 475 children were selected from the database, including 20 175 boys and 20 320 girls. The prevalence of simple abdominal obesity was 6.36% (2576/40 495), the prevalence of boys and girls was 7.41% (1494/20 175) and 5.32% (1082/20 320) respectively. The prevalence of combined obesity was 16.33% (6611/40 495), the prevalence of boys and girls was 21.30% (4298/20 175) and 11.38% (2313/20 320) respectively. There were statistical differences in the body type distribution of both boys and girls (χ(2) = 869.01, P < 0.01). The prevalence of high blood pressure was 9.62% (3896/40 495), the prevalence of boys (10.05% (2028/20 175)) was higher than girls (9.19% (1868/20 320)) (χ(2) = 8.59, P < 0.01). The value of SBP and DBP among combined obesity group (boys: (103.8 ± 11.3), (64.7 ± 10.1) mm Hg (1 mm Hg = 0.133 kPa); girls: (102.9 ± 12.1), (64.5 ± 10.0) mm Hg) > simple abdominal obesity group (boys: (99.5 ± 11.2), (61.6 ± 9.9) mm Hg; girls: (99.6 ± 11.4), (62.6 ± 9.3) mm Hg)> normal weight group (boys: (97.4 ± 10.8), (60.5 ± 9.4) mm Hg; girls: (97.2 ± 10.8), (60.8 ± 9.4) mm Hg), and the differences were statistical significant (Fboys: 113.22, 62.05; Fgirls: 54.19, 32.31, P < 0.01). According to the results of multilevel model, among boys, compared with the normal weight group, the SBP of combined obesity group and simple abdominal obesity group was 6.0 and 1.8 mm Hg higher respectively (Wald χ(2): 17.55, 204.94, P < 0.01); the DBP was 4.0 and 0.9 mm Hg higher respectively (Wald χ(2): 6.37, 114.05, P < 0.05). Among girls, the SBP was 5.0 and 2.1 mm Hg higher respectively (Wald χ(2):16.47, 92.52, P < 0.01); the DBP was 3.5 and 1.6 mm Hg higher respectively (Wald χ(2): 12.29, 57.52, P < 0.01). Comparing with normal group, the risk of high SBP among boys with simple abdominal obesity was higher (OR = 1.48; 95%CI: 1.06-2.06), and both the boys (SBP: OR = 3.06; 95%CI: 2.28-4.11) ; DBP: OR = 2.72; 95%CI: 1.99-3.72) and girls (SBP: OR = 2.48; 95%CI: 1.75-3.53; DBP: OR = 2.64; 95%CI: 1.82-3.93) in combined obesity group had a higher risk of high SBP and high DBP.

CONCLUSIONSimple abdominal obesity is associated with the increasing of blood pressure in children, and combined obesity has a closer ties than simple abdominal obesity.

Blood Pressure ; Body Mass Index ; Child ; China ; epidemiology ; Female ; Humans ; Hypertension ; epidemiology ; Male ; Obesity, Abdominal ; epidemiology ; Pediatric Obesity ; epidemiology

ObjectiveTo investigate the value of 99Tcm labeled survivin mRNA antisense peptide nucleic acid (PNA) as an imaging agent in the specific diagnosis for carcinoma.MethodsSurvivin mRNA antisense PNA was labeled directly with 99Tcm by the ligand-exchange method.Twenty nude mice with lung carcinoma A549 xenografts were randomly divided into 4 groups.Three groups were used for biodistribution study and one group was used for imaging study.Other twenty mice infected by staphylococcus aureus underwent the same procedure.The biodistribution and imaging of 99Tcm-survivin mRNA antisense PNA was studied at 1,2 and 4 h respectively after the intravenous injection in nude mice bearing lung carcinoma A549 xenografts or inflammation models.SPSS 13.0 was used in the study and all data were analyzed by t test.ResultsBiodistribution results showed that the highest radioactivity was found in the liver,and then in the kidney.Four hours after the administration of the imaging agent,the radioactivity ratios of target-tonon target (T/NT,tumor or inflamumatory lesions to the contralateral regions) in tumor model group were significantly higher than those in inflammation model group ( 3.69 ± 1.13 vs 2.03 ± 0.47,t =3.01,P =0.02 ).Tumors were clearly visible in the tumor model groups at 0.5 h and still clearly seen at 4 h after the injection of antisense PNA.On the contrary,inflammatory lesions could not be seen clearly.Conclusion 99Tcm labeled survivin mRNA antisense PNA can be used to distinguish tumor from inflammation and it may provide a new feasible method for specific tumor diagnosis.

OBJECTIVESTo explore the clinical and pathological features and the pathogenesis of primary biliary cirrhosis (PBC) in Chinese Mainland.

METHODS30 PBC patients were divided into the early group (Scheuer stage I and II, 19 patients) and the late group (Scheuer stage III and IV, 11 patients). The data of clinics and serology were analyzed, and the pathological features of the liver tissues were characterized. The changes of dendritic cells (DCs) and hepatic stellate cells (HSCs) were studied by immunohistochemistry.

RESULTSIn all the PBC patients, the rate of the male to the female was 1 to 5, and the average age was 40.6 years. The mean levels of TBiL, ALP and GGT in the sera were (95.9+-88.5) micromol/L, (537.2+-339.2) U/L, and (582.0+-351.2) U/L, respectively. 73.3% patients showed AMA positive, and the level of GGT was positively correlated with the AMA level according to the result of statistical analysis (r=0.778, P=0.000). The symptoms of jaundice and hepatomegaly were presented more commonly in the late group than those in the early group (chi2=5.182, P<0.05; chi2=13.659, P<0.01, respectively). The main changes of morphology of PBC located in portal tracts. The liver tissues in the early stage of PBC showed the damage of bile ducts and obvious proliferation of small bile ducts. The granulomas, the lymphoid follicles and the foamy cells were found in the liver tissues of PBC (2/19 patients, 12/19 patients, and 10/19 patients in the early stage respectively, while 0/11 patients, 4/11 patients, and 3/11 patients in the late stage respectively). There was significant difference between the early stage and the late stage in presence of the lymphoid follicles and the foamy cells (t=4.489, P<0.05; t=4.019, P<0.05, respectively). The biliary pigmentary particles were mainly accumulated in the liver cells around the portal tracts in 90.0% PBC patients, and the accumulation of copper and iron increased, compared with that in normal specimens. The DCs and HSCs located mainly in the portal tracts, especially around the damaged bile ducts.

CONCLUSIONSThere are some clinical and pathological characteristics in the patients with PBC. The level of AMA has no direct relationship with the level of transaminase or bilirubin. The proliferated bile ductules may express the antigens which maybe the target of immune attack. As an antigen-presenting cell, DCs may play an important role in the pathogenesis of PBC.

Adolescent ; Adult ; Antibodies, Antinuclear ; blood ; Antigen-Presenting Cells ; immunology ; pathology ; Dendritic Cells ; pathology ; Female ; Humans ; Liver ; pathology ; Liver Cirrhosis, Biliary ; etiology ; immunology ; pathology ; Male ; Middle Aged ; Mitochondria ; immunology

OBJECTIVETo develop human recombinant neutralizing IgG monoclonal antibodies to hepatitis A virus (HAV) by baculovirus expression system.

METHODSThe heavy and light chain genes of two human-derived neutralizing Fab antibodies to HAV were cloned into baculovirus expression vector Pac-kappa-Fc and Pac-L-Fc, and further expressed in insect cells as IgG antibodies. The IgG products were purified and well characterized.

RESULTSThe baculovirus expressed McAb HAFc16 fully retained the specificity of binding to hepatitis A virus and the competition with mouse anti-hepatitis A virus McAb using ELISA. The viral neutralization assay in vitro demonstrated the retention of antibody function after expression of the human antibody in insect cells. The other expressed antibody HAFc78 also has the neutralizing activity but it is directed against different epitopes of HAV when compared with HAFc16.

CONCLUSIONThe recombinant baculovirus/insect cells expressed human neutralizing IgG antibodies to hepatitis A virus retained all biological functions specific for hepatitis A virus. The results provided the possibility of using these antibodies to rapidly protect high risk or early exposure populations from hepatitis A virus infection.

Antibodies, Monoclonal ; biosynthesis ; immunology ; Baculoviridae ; genetics ; Hepatitis A virus ; immunology ; Hepatitis Antibodies ; biosynthesis ; immunology ; Humans ; Immunoglobulin Fab Fragments ; biosynthesis ; immunology ; Immunoglobulin G ; biosynthesis ; immunology ; Immunoglobulin Heavy Chains ; genetics ; Immunoglobulin Light Chains ; genetics ; Recombinant Proteins ; biosynthesis ; immunology

BACKGROUNDTo explore the pathological features and pathogenesis of severe acute respiratory syndrome (SARS) to provide evidence for the clinical treatment and prevention of SARS.

METHODSPathological features of 2 cases of full autopsy and 4 cases of needle biopsy tissue samples from the patients who died from SARS were studied by light and electron microscopy. The distribution and quantity of lymphocyte subpopulations in the lungs and immune organs from SARS patients were analyzed by immunohistochemistry. The location and semi-quantitative analysis of SARS coronavirus in the tissue specimens were studied by electron microscopy, in situ hybridization and immunohistochemistry.

RESULTSIn total of 6 cases, diffuse alveolar damage and alveolar cell proliferation were common. The major pathological changes of 2 autopsy cases of SARS in lung tissues were acute pulmonary interstitial and alveolar exudative inflammation, and 2 autopsy and one biopsy lung tissues showed alveolar hyaline membrane formation. Terminal bronchiolar and alveolar desquamation of lung tissues in one autopsy and 2 biopsy cases were noted. Among 6 cases, 2 biopsy cases presented early pulmonary fibrosis and alveolar organization. Meanwhile, the immune organs, including lymph nodes and spleens from 2 autopsy cases of SARS whose disease courses were less than 12 days showed extensive hemorrhagic necrosis, reactive macrophage/histocyte proliferation, with relative depression of mononuclear and granulocytic clones in the bone marrows. However, spleen and bone marrow biopsy tissue samples from 4 dead SARS cases whose clinical course lasted from 21 to 40 days presented repairing changes. SARS coronaviruses were mainly identified in type I and II alveolar epithelia, macrophages, and endothelia; meanwhile, some renal tubular epithelial cells, cardiomyocytes, mucosal and crypt epithelial cells of gastrointestinal tracts, parenchymal cells in adrenal glands, lymphocytes, testicular epithelial cells and Leydig's cells were also detected by electron microscopy combined with in situ hybridization. The semi-quantitative analysis of lymphocyte subpopulations revealed that the proportion of CD8+ T lymphocytes were about 80% of the total infiltrative inflammatory cells in the pulmonary interstitium, with a few CD4+ lymphocytes CD3+, CD4+, CD8+ or CD20+ lymphocyte subpopulations were obviously decreased and there was imbalance in number and proportion, while CD57+, CD68+, S-100+ and HLA-DR+ cells were relatively increased in lymph nodes and spleens.

CONCLUSIONSHistologically, the pulmonary changes could be divided into acute inflammatory exudative, terminal bronchiolar and alveolar desquamative and proliferative repair stages or types during the pathological process of SARS. SARS coronavirus was found in multi-target cells in vivo, which means that SARS coronavirus might cause multi-organ damages which were predominant in lungs. There were varying degrees of decrease and imbalance in number and proportion of lymphocyte subpopulations in the immune organs of the patients with SARS. However, these changes may be reversible. It was found that cellular immune responses were predominant in the lungs of SARS cases, which might play an important role in getting rid of coronaviruses in infected cells and inducing immune mediated injury.

Aged ; Female ; Humans ; Lung ; immunology ; pathology ; virology ; Lymphocyte Subsets ; immunology ; Male ; Middle Aged ; SARS Virus ; isolation & purification ; ultrastructure ; Severe Acute Respiratory Syndrome ; immunology ; pathology ; virology