ObjectiveTo study the relationship between C-erbB-2 and estrogen (ER) and progesterone (PR) receptors, and the relationship between C-erbB-2, ER, PR with histologic grade. MethodsTo detect ER, PR and C-erbB-2 states by using immunohistochemical analysis and fluorescence in situ hybridization for C-erbB-2 in 163 unselected invasive breast carcinomas. ResultsC-erbB-2, ER ,PR were expressed in 21.5％ ,64.4％ ,44.2％ of 163 cases respectivly . 5 pure mucinous carcinomas , 3 tubular carcinomas and 1 micropapillary carcinoma were ER + ( 100.0％ ) 、C-erbB-2 - ( 100.0％ ) and PR + (40.0％ ,66.7％, 100.0％ ). C-erbB-2 was positive in 22.3％ of grade Ⅱ and 27.0％ of grade Ⅲ invasive ductal carcinomas and negative in all grade Ⅰ invasive ductal carcinomas.ER and PR expression were decreased significantly in C-erbB-2 + tumors compared with C-erbB-2 - tumors( ER,25. 7％ vs 75.0％ ; PR,25.7％ vs 49.2％ ). Although ER or PR expression is decreased in C-erbB-2 + tumors, a substantial proportion of them still express ER or PR. ConclusionC-erbB-2 overexpression or amplifcation was limited to a minority of invasive breast carcinomas. Tumour grade was an independent predictor for ER expression. ER was expressed in small number of high-grade and in large number of grade Ⅰ invasive ductal carcinomas. C-erbB-2 overexpression or amplification essentially was limited to grades Ⅱ and Ⅲ ductal carcinomas and correlated inversely with ER or PR expression.

OBJECTIVETo introduce practical diagnostic criterion of blood stasis syndrome (BSS), and to evaluate its reliability and validity.

METHODSBy referring to three diagnostic criteria of BSS [practical diagnostic criterion of BSS (criterion A), diagnostic criterion of BSS in 1986 (criterion B), Consensus of Integrative Medicine on BSS Diagnosis in 2011 (criterion C)], 712 patients from different departments of Xiyuan Hospital were recruited. The reliability of criterion A and its consistency with the other two criteria were assessed using Kappa coefficient. A Bayesian approach was also employed to assess the sensitivity and specificity of criterion A.

RESULTSAccording to the consistency check, criterion A presented good consistency when used by different researchers (the diagnostic accordance rate was 91. 96%, Kappa =0. 82, P <0.001). Meanwhile, there was an acceptable diagnostic consistency among the three diagnostic criteria. Bayesian estimation suggested that criterion A had higher sensitivity but similar specificity, as compared with criterion B or criterion C. Compared with criterion B [the median of sensitivity and specificity were 0. 762 (95% Cl: 0. 731 -0. 790) and 0. 902 (95% Cl: 0. 858 -0. 936) respectively, the median of sensitivity and specificity of criterion A were 0. 911 (95% CI: 0. 888 - 0. 930) and 0. 875 (95% CI: 0. 826 - 0. 915) respectively. Estimating the difference between criterion A and B, the median of sensitivity and specificity were 0. 149 (95% CI: 0. 112 -0.184) and -0. 026 (95% CI:-0. 085 -0. 033) respectively. Compared with criterion C [the median of sensitivity and specificity were 0. 831 (95% Cl: 0. 804 -0. 857) and 0. 892 (95% CI: 0. 848 - 0. 926) respectively], the median of sensitivity and specificity of criterion A were 0. 912 (95% CI: 0. 889 -0. 932) and 0. 880 (95%CI: 0. 833 - 0.919) respectively. Estimating the difference between criterion A and C, the median of sensitivity and specificity were 0. 081 (95% CI: 0.047 - 0.114) and -0.011 (95%CI: -0.070 -0.046) respectively.

CONCLUSIONCompared with criterion B and C, criterion A not only had better reliability, but also could significantly improve the sensitivity without obviously lowering the specificity.

Bayes Theorem ; Consensus ; Hematologic Diseases ; diagnosis ; Humans ; Medicine, Chinese Traditional ; Reproducibility of Results ; Sensitivity and Specificity

The key points for better protection of trehalose in freeze-drying red blood cells (RBCs) are to resolve non-osmosis of trehalose to red blood cells and to make cytoplasmic trehalose to reach effective concentration. This study was aimed to investigate the regularity of loading RBCs with trehalose, screen out optimal loading condition and evaluate the effect of trehalose on physico-chemical parameters of RBCs during the period of loading. The cytoplasmic trehalose concentration in red blood cells, free hemoglobin and ATP level were determined at different incubation temperatures (4, 22 and 37 degrees C), different trehaolse concentrations (0, 200, 400, 600, 800 and 1000 mmol/L) and different incubation times (2, 4, 6, 8 and 10 hours), the cytoplasmic trehalose, free hemoglobin (FHb), hemoglobin (Hb) and mean corpuscular volume (MCV) in fresh RBCs and RBCs stored for 72 hours at 4 degrees C were compared, when loading condition was ensured. The results showed that with increase of incubation temperature, time and extracellular trehalose concentration, the loading of trehalose in RBCs also increased. Under the optimal loading condition, cytoplasmic trehalose concentration and free hemoglobin level of fresh RBCs and RBCs stored for 72 hours at 4 degrees C were 65.505 +/- 6.314 mmol/L, 66.2 +/- 5.002 mmol/L and 6.567 +/- 2.568 g/L, 16.168 +/- 3.922 g/L respectively. It is concluded that the most optimal condition of loading trehalose is that fresh RBCs incubate in 800 mmol/L trehalose solution for 8 hours at 37 degrees C. This condition can result in a efficient cytoplasmic trehalose concentration. The study provides an important basis for long-term preservation of RBCs.
Biological Transport, Active ; drug effects ; Blood Preservation ; methods ; Cryopreservation ; methods ; Cryoprotective Agents ; metabolism ; pharmacology ; Erythrocyte Membrane ; metabolism ; Erythrocytes ; Freeze Drying ; Humans ; Osmotic Fragility ; Temperature ; Time Factors ; Trehalose ; metabolism ; pharmacology

The objective of this study was to investigate the effect of different rehydration conditions on recovery of the lyophilized red blood cells (RBC) so as to optimize the RBC rehydration. The different conditions, including different rehydration solution, the rehydration temperature, volume change rate of the lyophilized RBC rehydrated by the vapor firstly, were studied, the recovery rate and change of physiological and biochemical properties of the rehydrated RBC were detected. The results indicated that the solution of 10% (w/v) PVP40 in PBS showed the best effect, and the RBC recovery rate increased with increasing of rehydration temperature, and the optimal temperature of rehydration was at 37 degrees C. Pre-rehydration in condition of vapor could raise the RBC recovery rate, and promote the MCV and RDW to close to index of the fresh RBC, the deformability of the rehydrated RBC was no serious as compared with RBC preserved in conventional condition, but the activity level of ATP, G-6-PD, SOD, 2, 3-DPG of the rehydrated RBC less decreased. It is concluded that the optimal rehydration conditions for lyophilized RBC are pre-rehydration in the 37 degrees C with vapor firstly, PBS + 10% (w/v) PVP40 rehydration solution and rehydration temperature at 37 degrees C, but the protection of RBC membrane needs to be furtherly studied.
Blood Preservation ; methods ; Erythrocyte Count ; Erythrocytes ; Freeze Drying ; methods ; Humans ; Rehydration Solutions ; Temperature

OBJECTIVETo construct E1-deletion and replication-defective human type 5 recombinant adenovirus vector and to study the effect of p16INK4a on proliferation and aging of A549 cells.

METHODSp16INK4a cDNA was cloned into pAdCMV to construct recombinant pAdCMV p16INK4a, which was co-transfected into 293 cell together with pJM17. The recombinant p16INK4a adenovirus (Ad-p16INK4a) was generated by homologous recombination and identified with duplex PCR. Lung cancer cell A549, which has a homozygous deletion of p16INK4a gene, was infected with the prepared Ad-p16INK4a virus. X-gal staining and TRAP-ELISA were used for detecting senescence-associated beta-galactosidase and telomerase activities in A549 cells.

RESULTSImmunohistochemical staining and Western blot indicated that p16INK4a gene was transferred into A549 cell with more than 95% efficiency by recombinant adenovirus and p16INK4a protein was expressed at a high level- p16INK4a could markedly inhibit growth of A549 cells, induced expression of senescence-associated beta-galactosidase and suppressed telomerase activity in A549 cells.

CONCLUSIONRecombinant adenovirus vector could efficiently mediate transfer and expression of foreign genes in human cell and could be used for gene immunization and gene therapy; p16INK4a could inhibit A549 cell growth and induce its replicative senescence.

Adenoviridae ; genetics ; Cell Line, Tumor ; Cell Proliferation ; Cellular Senescence ; Cyclin-Dependent Kinase Inhibitor p16 ; biosynthesis ; genetics ; physiology ; Gene Expression ; Genetic Vectors ; Humans ; Recombination, Genetic ; Transfection

This study was purposed to investigate the effect of different compositions and concentrations of lyophilizing protectants on recovery of RBCs and hemoglobin (Hb) after rehydration of lyophilized RBCs. The RBC lyophilizing protectants composed of a series concentrations of PVP, trehalose and different osmotic protectants were applied for protecting lyophilizing process of RBCs, the recovery of RBCs and Hb after rehydration of lyophilized RBCs was detected. The results showed that there were significant differences in loss ratio of RBCs between protectants composed of different compositions and concentrations (p<0.05 or p<0.01). The loss ratio of RBCs in protectant containing 30% PVP40, 150 mmol/L trehalose and 2% BSA was minimum (0.02%), the loss ratio of RBCs in protectant containing 6% PVP 360, 100 mmol/L trehalose and 2% BSA was maximum (0.27%). The difference of effect between 150 and 50 mmol/L trehalose was statistically significant (p<0.01). The recovery rates of RBCs and Hb in protectants contained PVP40 of different concentrations were different after rehydration of lyophilized RBCs. The protectant containing 15% PVP40, 150 mmol/L trehalose and 2% BSA showed optimal protective efficacy for lyophilized RBCs, the recovery rates of RBCs and Hb were 61.29+/-4.11% and 62.49+/-5.91% respectively, which were statistically different from other protectants (p<0.01). The protectants containing glycerol displayed best efficiency in lyophilization too, the recovery rates of RBCs and Hb were 65.97+/-4.52% and 67.24+/-5.94%, respectively. It is concluded that the protectants composed of 0.8 mol/L glycerol, 15% PVP40, 150 mmol/L trehalose and 2% BSA (pH 7.3 ) may be used as the protectant lyophilizing human RBCs in future study.
Blood Preservation ; methods ; Cryoprotective Agents ; administration & dosage ; analysis ; Erythrocytes ; Freeze Drying ; methods ; Humans ; Trehalose ; administration & dosage ; analysis

Objective To study the clinical characteristics and genetic features of SPG4 gene in a family with hereditary spastic paraplegia (HSP). Methods The four patients from one LinYi family were clinically diagnosed as having HSP according to Harding's criteria and their peripheral blood samples were collected. We typed the short tandem repeat (STR) loci closely connected with the known HSP cause gane locus at physical distance and genetic linkage analysis was performed on them. Their haplotypes were structured and then screening of gene mutations was performed. Results Non-elimination of linkage was found between D2S2351 and D2S2255 and cause gene, and the LOD scores in other locus were negative value and eliminated the linkage, which implied that the location was in the ADHSP locus of chromosome 2p22 (SPG4) and the candidate gene was spastin gene. Screening of gene mutations found that the mutation loci lied in heterozygous A and G at nucleotide 1168 in spostin gene. The symptoms of the patients manifested as stiffness, instability or weakness of the legs. Conclusions The patients in this family have typical clinical symptoms of HSP, mainly resulting from the novel mutation (spastin: c1168 A>G).

OBJECTIVETo evaluate chest radiographic findings of children with 2009 influenza (H1N1) virus infection.

METHODData of 235 patients who had microbiologically confirmed H1N1 infection and available chest radiograph obtained between May 1(st) 2009 and Jan. 31(st) 2010 were retrospectively analyzed. The final study group was divided on the basis of clinical course [group 1 mild, outpatients without hospitalization (n = 172); group 2 moderate, inpatients with brief hospitalization (n = 49); group 3 severe, ICU admission (n = 14)]. Four pediatric radiologists reviewed all the chest radiographs of lung parenchyma, airway, pleural abnormalities and also anatomic distribution of the disease.

RESULTNo significant sex or age differences were found among the study groups (P > 0.05). The mean interval between the onset of clinical symptom and the initial chest radiography was (5.91 ± 1.64) days (group 1), (3.60 ± 1.43) days (group 2) and (1.21 ± 0.41) days (group 3), respectively. The differences among the three groups were significant statistically (χ(2) = 13.368, P < 0.01). The ratio of abnormality presented at initial chest X-ray was 79.7% in group 1, 91.8% in group 2 and 100% in group 3. Radiographically, there were prominent peribronchial markings (group 1, 55.2%; group 2, 83.7%; and group 3, 78.6%), consolidation (group 1, 34.3%; group 2, 69.4%; and group 3, 100.0%), hyperinflation (group 1, 22.1%; group 2, 44.9%; and group 3, 50.0%) and ground glass opacity (group 1, 0.6%; group 2, 2.0%; and group 3, 14.3%) in the chest radiographs. The differences of presenting were statistically significant (P < 0.01). In the severe group, the lesions distributed diffusely and asymmetrically with multi-lobe involvements.

CONCLUSIONIn children with 2009 influenza A H1N1 viral infection, the interval between the onset of clinical symptom and initial chest radiography, the ratio of abnormality presented at initial chest X-ray film and the severity of chest film are parallel to their clinical situation.

Child ; Child, Preschool ; Female ; Humans ; Infant ; Infant, Newborn ; Influenza A Virus, H1N1 Subtype ; Influenza, Human ; diagnostic imaging ; virology ; Male ; Retrospective Studies ; Tomography, X-Ray Computed

OBJECTIVETo evaluate the protective effect of ischemic preconditioning against cold ischemia and reperfusion injury of rat intestinal graft following orthotopic transplantation.

METHODSEighty SD rats were randomly assigned into two groups with and without ischemic preconditioning, and each group was divided into 4 subgroups (n=10) according to the intestinal graft cold ischemia time of 3, 6, 12, and 18 h, respectively. Ischemic preconditioning model was established, and the small intestinal graft was preserved at 4 degrees celsius; in Ringer lactate solution for the corresponding time, followed by orthotopic transplantation of the graft. The graft samples were collected for histological examination 1 h after reperfusion, and nuclear factor-kappaB (NF-kappaB) expression in the epithelial cells was detected.

RESULTSIschemia preconditioning obviously relieved the histological ischemia/reperfusion injury, as shown by regular alignment of the small intestinal villi, alleviated muscular layer edema and decreased expression of NF-kappaB in the epithelia of the graft in groups with cold preservation.

CONCLUSIONIschemic preconditioning can protect the intestinal graft from cold ischemia/reperfusion injury, and NF-kappaB is an important cytokine in ischemia preconditioning.

Animals ; Cold Ischemia ; Intestine, Small ; pathology ; transplantation ; Ischemic Preconditioning ; NF-kappa B ; metabolism ; Organ Preservation ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; prevention & control

This study was aimed to investigate the aggregation of rehydrated-lyophilized platelets. The aggregation rate of fresh and rehydrated-lyophilized platelets were measured by using thrombin, ristocetin, ADP and collagen as inductors and APACT2 aggregameter; the effects of intra- and extra-cellular trehalose on maximum aggregation rate of rehydrated-lyophilized platelets were detected by using ADP as an inductor. The results showed that the aggregation rate of fresh platelets was all about 100%, while aggregation rate of rehydrated lyophilized platelets was (70.17 +/- 7.36)%, (15.3 +/- 2.81)%, (68.67 +/- 6.86)%, (64.67 +/- 11.6)% respectively, when the concentration of thrombin, ristocetin, ADP and collagen was 1 U/ml, 1.6 mg/ml, 20 micromol/L and 2 microg/ml. The maximum aggregation rates of rehydrated-lyophilized platelets in intra- and extra-cellular trehalose, extracellular trehalose and blank control groups were (66.0 +/- 4.69)%, (25.3 +/- 2.42)% and (11.5 +/- 1.87)% (P < 0.01), meanwhile there was significant difference of rehydrated-lyophilized platelet aggregation rate between intra- and extra-cellular trehalose and extracellular trehalose groups (P < 0.01). It is concluded that the concentrations of thrombin (1 U/ml), ristocetin (1.6 mg/ml), ADP (20 micromol/L) and collagen (2 microg/ml) are optimal for platelets aggregation tests, the internal and extracellular trehalose significantly enhance the aggregation of rehydrated-lyophilized platelets.
Blood Platelets ; cytology ; drug effects ; metabolism ; Blood Preservation ; methods ; Freeze Drying ; methods ; Humans ; Platelet Aggregation ; physiology