To fill the urgent need of researches on the structure and function of proteins to obtain conformation information, we combined targeted molecular dynamics ( TMD) simulation with protein network methods to analyze conformation variations in the activation process of β2 adrenergic receptor. First, targeted MD was used to obtain the conformation resembles in the activation process, and then the protein network method was applied to identify the key residues and pathways in the activation process. The results indicate that the activa-tion process of β2 adrenergic receptor involves in the cooperation of all regions and the connector region of transmembrane helices is signaling hubs. In addition, the helix ends, including intracellular and extracellular loops, are the core areas. The pathway analysis reveals that there is more than one signaling pathway. All the pathways start from Ser204 of the ligand pocket and finally transmit to NPxxY or ICL2 region, which are depended on the different pathways. While the helix TMIII, TMV, TMVI are the important areas in all the pathways. The observations from the work provide valuable information at molecular level for unraveling the signal transduction mechanism associated with the activation process.

Adult ; Antibodies, Viral ; blood ; China ; epidemiology ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Immunoglobulin G ; blood ; Rubella ; epidemiology ; Rubella virus ; immunology ; isolation & purification ; Seroepidemiologic Studies

Objective To investigate the diagnostic value of dual-energy CT pulmonary angiography (DE-CTPA) and dual-energy CT lung perfusion (DE-CTLP) in the assessment of pulmonary embolism comparing with pulmonary ventilation-perfusion scintigraphy. Methods Fifty patients suspected of PE (26 males, 24 females) underwent both DE-CTPA, DE-CTLP and pulmonary ventilation-perfusion scintigraphy. The results were compared and the correlation between the intravascular clots in DE-CTPA and the perfusion defects in DE-CTLP was analyzed. The sensitivity, specificity and accuracy of DE-CTLP for PE were assessed according to scintigraphic results which was considered as a "gold" standard. Result ( 1 ) Of 50 cases,920 segments in 46 cases were diagnostable, 4 cases were not included because of the poor image quality. lntravascular clots were found in 262 segments on DE-CTPA and perfusion defects in 266 segments were identified on DE-CTLP, while 268 segments were positive on scintigraphy. (2) The perfusion defects on DE-CTLP were correlated well with the clots on CTPA ( r = 0. 883, P < 0. 01 ). The Dual energy CT Lung perfusion imaging had a good consistence with scintigraphy (Kappa = 0. 940, P <0. 01 ). The sensitivity, specificity, positive and negative predictive value were 96. 2% (279/290), 98.0%(641/654), 95. 5% (279/292) and 98. 3% (641/652), respectively. (3)The radiation dose of DE-CTPA and CE-CTLP scan was(4. 37 ± 0. 47) mSv by using CareDose 4D technique. Conclusion DE-CTPA and CE-CTLP can provide pulmonary vascular morphology and parenchyma perfusion information which are useful and valuable for the diagnosis of PE.

OBJECTIVETo evaluate the effects of mechanical ventilation on pulmonary function during short duration of general anesthesia with tracheal intubation, and assess the safety of controlled spontaneous respiration during general anesthesia.

METHODSFifty-three adult patients (aged 18-55 years, ASA physical status I-II) scheduled for elective unilateral tympanoplasty were randomly assigned into mechanical ventilation group (group M, n=28) and spontaneous respiration group (group S, n=25). Anesthesia induction was performed in group M with intravenous propofol (2 mg/kg), fentanyl (3 microg

RESULTSA total of 43 patients (group M, n=23; group S, n=20) were included in the study with 10 dropouts due to failed attempt to obtain arterial blood samples (8 patients) or severe bucking during intubation (2 patients). No significant differences were found in HR and MAP between the two groups (P>0.05). The pH and SpO(2) [ (97.9-/+1.00)% at the lowest] and PaO(2) in group S were significantly lower and the PaCO(2) was higher than those in group M (P<0.05). In group S, the pH values were 7.274-/+0.025 and 7.331-/+0.039, PaCO(2) values were 60-/+6 and 53-/+5 mmHg, and PETCO(2) values were 53-/+ 6 and 48-/+7 mmHg, and the PaO(2) values were 143-/+37 and 165-/+49 mmHg immediately and 150 min after the intubation, respectively. These values were considered safe under the concept of permissive hypercapnia. No significant differences were found in the P(A-a)DO(2), RI, VD/VT and TFC between or within the two groups (P>0.05), nor were moving, bucking, swallowing and awareness recorded during the surgical procedures.

CONCLUSIONIn essentially normal lungs, short-term mechanical ventilation during general anesthesia with tracheal intubation does not damage the lung functions, and spontaneous respiration can offer sufficient oxygen supply without causing harmful carbon dioxide retention.

Adolescent ; Adult ; Anesthesia, General ; methods ; Female ; Humans ; Intubation, Intratracheal ; Lung ; physiology ; Male ; Middle Aged ; Respiration ; Respiration, Artificial ; methods ; Tympanoplasty ; methods ; Young Adult

In order to observe the proliferative and apoptotic situation of megakaryocytes in patients with myelodysplastic syndromes (MDS). CD41 immunoenzyme labeling (alkaline phosphatase anti-alkaline phosphatase APAAP)/DNA in situ end labelling (ISEL) double stained techniques was used onto plastic cold embedded bone marrow sections in 29 MDS patients to analyse the proliferative and apoptostic characterization of megakaryocytic line with 14 cases of iron deficient diseases (IDA) as control. The results showed that the mean CD41 positive cell number in MDS group was (26.23 +/- 8.18) /mm(2) with a count of (15.64 +/- 7.11) /mm(2) in control group (p < 0.05). The small-micro megakaryocytes in MDS is much higher than that in IDA group (P<0.01). There was a positive co-relation between total megakaryocytes and small-micro megakaryocytes count in MDS (r = 0.702, p<0.01). Some megakaryocytes distributed abnormally around trabecula and formed small or large clusters. Apoptotic megakaryocytes in MDS occupied 4.40% and 9.32% of all CD14 positive cells and all apoptotic cells respectively (p > 0.5 comparing with control). Apoptosis in megakargocytic line occurred only in small-micro megakaryocytes and showed positive co-relation to the number of micro-megakaryocytes. Some apoptotic cell with morphologic characters of megakaryocytes expressed no CD41. It is concluded that overproliferation of megakaryocytes exists in MDS. Apoptosis occurring in micro-megakaryocytes may be a kind of physiological response to abnormal megakaryopoicsis in MDS. No obvious increased apoptosis of megakaryocytes in MDS was found perhaps due to lack of surface antigens CD41 in some later stages of apoptotic cell.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Apoptosis ; Cell Division ; Child ; Female ; Humans ; Male ; Megakaryocytes ; pathology ; Middle Aged ; Myelodysplastic Syndromes ; pathology

OBJECTIVETo investigate the total in situ apoptotic cell number and the apoptotic situation in erythroid cell and megakaryocytes in patients with myelodysplastic syndromes (MDS).

METHODSApoptosis cell number and the apoptotic situation of erythroid cell and megakaryocytes were analysed on cold embedded bone marrow sections from 25 MDS patients by DNA in situ end labelling (ISEL)/alkaline phosphatase anti-alkaline phosphatase (APAAP) double stained techniques. Fourteen cases of iron deficiency anemia (IDA) were taken as control.

RESULTSMean apoptotic cell numbers in MDS and control group were (39.44 +/- 29.34)/mm(2) and (13.43 +/- 8.39)/mm(2) respectively (P < 0.01). RA/RAS subtypes had a higher apoptosis ratio (47.56 +/- 32.86/mm(2)) than that in RAEB/RAEB-t subtypes (21.87 +/- 13.65/mm(2)) (P < 0.05). Double staining showed similar apoptosis percentage in erythroid cell and megakaryocytes in MDS patients comparing with that of controls (P > 0.05). Some apoptotic cells showing erythroid or megakaryocytic morphologic characteristics expressed no cluster differentiation antigen.

CONCLUSIONOverapoptosis existed in MDS, RA/RAS group had a higher apoptosis ratio than RAEB/RAEB-t group. No obvious increased apoptosis in erythroid cell and megakaryocytes was observed in MDS perhaps due to the loss of surface antigens in later stages of apoptotic cells.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Alkaline Phosphatase ; immunology ; Apoptosis ; Bone Marrow Cells ; cytology ; metabolism ; Child ; DNA ; genetics ; Female ; Humans ; Immunoenzyme Techniques ; In Situ Nick-End Labeling ; Male ; Middle Aged ; Myelodysplastic Syndromes ; genetics ; pathology

Based on a pair of specific primers, a 804-bp fragment was amplified from the plasmid pT-Cap containing Cap gene of Porcine Getah Virus(PGETV) and cloned into the prokaryotic expression vector pCold I which carried the His tag, this recombinant plasmid was then determined by enzyme digestion, PCR and DNA sequencing. This recombinant plasmid pCold-Cap was transformed into E. coli Rosetta 2, and PGETV Cap fusion protein was expressed through IPTG induction. The results showed that the Cap gene obtained efficient and soluble expression in Rosetta 2 induced by 0. Immol/L IPTG under 15"C for 24h, the expression quantity was 40. 2%. The product had a molecular mass about 32. 3kD as expected. The target protein was separated in gel slices and used to immunize Balb/c mice. The polyclonal antibody with high titer against Cap protein specifically analyzed by Western blot was obtained. The successful preparation of the polyclonal antibody laid the foundation for the further study on the detection and identification of PGETV.
Alphavirus ; genetics ; immunology ; metabolism ; Alphavirus Infections ; immunology ; veterinary ; virology ; Animals ; Antibodies, Viral ; blood ; immunology ; Blotting, Western ; Capsid Proteins ; genetics ; immunology ; isolation & purification ; metabolism ; DNA Primers ; genetics ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; Genetic Vectors ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Plasmids ; genetics ; Polymerase Chain Reaction ; Recombinant Fusion Proteins ; Swine ; Swine Diseases ; immunology ; virology ; Zoonoses

OBJECTIVETo investigate the apoptotic situation of CD(34) positive cells in myelodysplastic syndromes (MDS).

METHODIn 36 MDS patients, immunocytochemical technique was used for the detection of the expression of CD(34) antigen and DNA in situ end labelling (ISEL) (fluorescein) for the apoptotic signals. Fourteen cases of iron deficiency anemias (IDA) were used as controls.

RESULTS(1) CD(34) expression in MDS group was much higher than that in controls (49.2 +/- 38.5 vs 10.2 +/- 9.7, P < 0.01), and MDS cases had an obviously higher apoptotic rate than control did (69.1 +/- 28.2 vs 17.8 +/- 11.2, P < 0.01). (2) Expression of CD(34) was higher in transforming group (P < 0.05) than in non-transforming and post-transforming groups. Apoptotic rates in both non-transforming/transforming group were higher than in post-transforming group (P < 0.02 and < 0.05 respectively). (3) No apoptosis was found in CD(34) positive cells in MDS; (4) Both CD(34) positive cells and apoptotic cells formed into small or large clusters but did not co-distributed in a given area.

CONCLUSIONThere is overexpression of CD(34) antigen on hematopoietic cells in MDS. High CD(34) expression accompanied high apoptosis coexisted in the process of transformation from MDS to AML. Apoptosis-resistance of these CD(34) positive cells suggested that they came from malignant hematopoietic cell clones.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Antigens, CD34 ; metabolism ; Apoptosis ; Bone Marrow Cells ; immunology ; pathology ; Child ; Female ; Humans ; Immunoenzyme Techniques ; In Situ Nick-End Labeling ; Male ; Middle Aged ; Myelodysplastic Syndromes ; immunology ; pathology

OBJECTIVEAt present there is no effective therapeutic approach for stage IV neuroblastoma. We report our experience with allogenic hematopoietic stem cell transplantation as a means of treating this disorder in one child.

METHODSA 7-year-old boy with stage IV neuroblastoma received allogenetic hematopoietic stem cell transplantation. The donor was his mother who was haploid HLA-matched to the patient. Conditioning regimen consisted of fludarabin and melphalan. Stem cells were collected from peripheral blood and bone marrow of the donor.

RESULTSThe patient achieved hematopoietic reconstruction and was converted to full donor chimerism according to short tandem repeat sequence-polymerase chain reaction detection. The patient's neutrophil count recovered to more than 0.5 x 10(9)/L 10 days after transplantation. The patient's platelet count recovered to more than 20 x 10(9)/L 11 days after transplantation. Acute graft versus host disease occurred 8 days after transplantation and was improved after treatment. The patient survived in a 210-day-follow-up.

CONCLUSIONSHaploid HLA-matched allogeneic hematopoietic stem cell transplantation from parent donor was an alternative, safe and effective treatment for children with stage IV neuroblastoma.

Child ; Follow-Up Studies ; Graft vs Host Disease ; drug therapy ; Hematopoiesis ; Hematopoietic Stem Cell Transplantation ; Humans ; Male ; Neoplasm Staging ; Neuroblastoma ; pathology ; therapy ; Transplantation, Homologous

OBJECTIVETo analyse related risk factors of classic Kaposi' s sarcoma in Xinjiang.

METHODSA 1:4 case-control study was used and the conditional logistic regression model was performed in this study. Cases were followed up in Xinjiang while controls were selected by the same sex, nation and age within 5 years with cases.

RESULTSInterleukin-6,vascular endothelial growth factor, beta-MG, neopterin, human herpevirus 8, were found to be associated with Kaposi's sarcoma risk in one-way variance model while beta2 -MG and human herpevirus 8 entered the multiple conditional logistic regression model, and their ORs were 1.002(95%CI: 1.000-1.003), 81.041 (95%CI: 1.790-3669.620).

CONCLUSIONThere was a correlate relationship between beta2 -MG and classic Kaposi's sarcoma being found that human herpevirus 8 exposure related factors seemed to have played important roles on classic Kaposi's sarcoma in Xinjiang.

Adult ; Aged ; Aged, 80 and over ; Case-Control Studies ; China ; Female ; Herpesviridae Infections ; complications ; Humans ; Logistic Models ; Male ; Middle Aged ; Multivariate Analysis ; Risk Factors ; Sarcoma, Kaposi ; complications ; epidemiology ; metabolism