BACKGROUND:Large-scale expansion of undifferentiated and multipotential adipose-derived stem cells using serum-free culture system is a difficult issue to be resolved. OBJECTIVE:To establish an in vitro culture system combined with the extracellular matrix in order to investigate the efficiency, effectiveness and security of extracellular matrix on expanding adipose-derived stem cells. METHODS:In vitro isolated adipose-derived stem cells were seeded in traditional two-dimensional plastic plates and extracellular matrix-coated plates supplemented with serum-free medium respectively. After in vitro expansion, total cellnumber, expression of cellsurface markers, cellsenescence degree and multipotent differentiation ability (adipogenic, osteoblastic and chondrogenic differentiation) of adipose-derived stem cells cultured under both conditions were detected and compared. Moreover, the clinical safety of adipose-derived stem cells expanded in extracellular matrix-coated plates was investigated. RESULTS AND CONCLUSION:Total cellnumber of passage 5 adipose-derived stem cells cultured in extracellular matrix-coated plates was 10 times more than that in traditional two-dimensional plastic plates. Flow-cytometric analysis showed that adipose-derived stem cells cultured with extracellular matrix expressed stem cellsurface markers. cellular senescence examination showed that almost al of passage 15 adipose-derived stem cells cultured with extracellular matrix showed no aging, while most passage 5 adipose-derived stem cells cultured by the two-dimensional system aged and lost their proliferation ability. Multidirectional induction of adipose-derived stem cells showed that passage 15 adipose-derived stem cells cultured with extracellular matrix could stil differentiate into adipocytes, osteoblasts and chondrocytes as passage 5 adipose-derived stem cells did, which performed much better than the induced differentiations of passage 5 adipose-derived stem cells cultured by the two-dimensional system. Karyotype analysis and in vivo invasion experiment insured the clinical safety of adipose-derived stem cells expanded with extracellular matrix. Al above results suggest a safe and more efficient expansion system of extracellular matrix for clinical application using the serum-free culture system combined with extracellular matrix.

Mean hemoglobin (Hb) concentration of about 3 500 subjects derived from 17 studies of Himalayan highlanders (Tibetans, Sherpas, and Ladakhis) was compared with lowlanders (Chinese Han, Indian Tamils) lived in the Himalayas, and European climbers during Everest expeditions as well as Andean natives. The results found that Hb concentration in Himalayan highlanders was systemically lower than those reported for Andean natives and lowland immigrants. These comparative data demonstrated that a healthy native population may successfully reside at high altitude without a significant elevation in Hb, and the lower Hb levels of Himalayan highlanders than those of migrated lowlanders and Andean natives are an example of favourable adaptation over the generations. In addition, excessive polycythemia has frequently been used as a marker of chronic mountain sickness (CMS). Altitude populations who have a higher Hb concentration also have a higher incidence of CMS. The low Hb in Himalayans suggested as showing adaptation over many generations in Tibetan stock. Recent work in Tibet, suggested that Tibetans there may have adapted to high altitude as a result of evolutionary pressure selecting for genes which give an advantage at altitude. All of the population genomic and statistical analysis indicated that EPAS1 and EGLN1 are mostly likely responsible for high altitude adaptation and closely related to low Hb concentration in Tibetans. These data supported the hypothesis that Himalayan highlanders have evolved a genetically different erythropoietic response to chronic hypoxia by virtue of their much longer exposure to high altitude.
Adaptation, Physiological ; Altitude ; Asian Continental Ancestry Group ; genetics ; Basic Helix-Loop-Helix Transcription Factors ; genetics ; Evolution, Molecular ; Hemoglobins ; genetics ; Humans ; Hypoxia-Inducible Factor-Proline Dioxygenases ; genetics ; Tibet

OBJECTIVETo examine the effect of trastuzumab on cell proliferation, colony formation and changes of HER-2 proteins in human breast cancer cell line SKBR3 and human ovarian cancer cell line SKOV3 cells which overexpress p185 HER-2 but shed high or low HER-2 extracellular domain (ECD) levels.

METHODSSKBR3 cells and SKOV3 cells were treated with or without trastuzumab. Cell number and the rate of colony formation were calculated. Western blot analysis was used to detect p185 HER-2, HER-2 ECD and phospho-HER-2. Two-site ELISA assay was used for the detection of HER-2 ECD.

RESULTSTrastuzumab inhibited cell proliferation, colony formation, and decreased or eliminated the levels of two uncharacterized phospho-proteins (molar weight about 90 000 and 40 000) in SKBR3 cells shedding high level of HER-2 ECD expression. These responses were not observed in SKOV3 cells shedding low level of HER-2 ECD expression. But total p185, phospho-p185 and phospho-p95 proteins did not appear to change in SKBR3 and SKOV3 cells after treatment with trastuzumab. Trastuzumab reacts not only with proteolytic cleavage HER-2 ECD containing HER-2 ECD I , II , III and IV subdomains of p185 HER-2 extracellular domain, but also with the secreted autoinhibitor p68/ECD III a specifying 340 residues, identical to subdomains I and II from the extracellular domain of p185 HER-2, followed by a unique C-terminal sequence of 79 aa encoded by intron 8, which suggested that there may be a trastuzumab binding site on p68/ECD III a protein. Comparing with HER-2 ECD levels of the same number of SKBR3 cells, there was no significant decrease of HER-2 ECD shedding level after treatment with or without trastuzumab for 4 days in serum-free medium.

CONCLUSIONAntitumor effects of trastuzumab may be related to the two uncharacterized phospho-p90 and/or phospho-p40 proteins. There is probably a trastuzumab epitope on p68/ECD III a. The decrease of HER-2 ECD levels may be positively correlated with the number of SKBR3 cells.

Antibodies, Monoclonal ; pharmacology ; Antibodies, Monoclonal, Humanized ; Antineoplastic Agents ; pharmacology ; Blotting, Western ; Breast Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Ovarian Neoplasms ; metabolism ; pathology ; Phosphorylation ; Receptor, ErbB-2 ; metabolism ; Trastuzumab

Numerous genetic abnormalities which can not be identified by cytogenetic detection (e.g., gene mutations, gene expression abnormalities) have been gradually found, which means that the further molecular classification of AML (acute myeloid leukemia) with distinctive prognosis have arrived. For example, mutations of the transcription factor (CCAAT enhancer binding factor alpha, C/EBPalpha) or nucleophosmin-1 (NPM1) may predict better prognosis, whereas partial tandem duplications of the MLL gene (MLL-PTD), internal tandem duplications of FLT3 (FLT3-ITD) or mutations of WT1 gene confer worse prognosis. This review focuses on the features and relationship of these genetic abnormalities, as well as their influence on the prognosis of AML.
Humans ; Leukemia, Myeloid, Acute ; diagnosis ; genetics ; Prognosis

OBJECTIVEAim of our study was to compare hematological parameters in Tibetan natives with those in Han migrants living on the Tibet plateau in order to determine the potential effects of age, gender, and ethnicity on hematological response to hypoxia.

METHODSBlood hemoglobin (Hb, g/dl), hematocrit (Hct, %), red blood cells (RBC,10(6)/mm3) were measured in 3 588 healthy Tibetan natives and 3 371 Han migrants ranging in age from 5 to 72 years, living at a mean altitudes of 2 664 m, 3 813 m, 4 525m and 5 226 m.

RESULTSHemoglobin (Hb) concentration analysis was made by multiple regression equations relating hemoglobin to altitude and age. For 2 093 Han males, Hb = 9.612+ 0.001440xaltitude+ 0.06148xage. For 1 948 Tibetan males, Hb =12.202+ 0.000462xaltitude+ 0.02893xage. For 1 278 Han females, Hb = 10.858+ 0.000939xaltitude+ 0.02632xage. For 1 640 Tibetan females, Hb = 11.402+ 0.000626xaltitude+ 0.00412xage. Each of the four equations was statistically significant (P < 0.001), and had variance (r2) of 0.86 or more, indicating that altitude and age accounted for at least 85% of the variation in hemoglobin levels. The coefficients for altitude and for age were higher (P < 0.05) in Han males than in Tibetan males and higher (P < 0.05) in Han females than in Tibetan females. The Tibetan postmenopausal females had higher Hb values than premenopausal females only presented at altitude above 4 000 m while this phenomenon was beginning at altitude of 2 664 m among Han females.

CONCLUSIONWe conclude that gender and increasing age in Tibetans are associated with lower hemoglobin values than those in Han at high altitude, and we speculate that genetic factors seems to be important.

Adolescent ; Adult ; Aged ; Altitude ; Asian Continental Ancestry Group ; Child ; Child, Preschool ; Ethnic Groups ; Female ; Hematocrit ; Hemoglobins ; analysis ; Humans ; Hypoxia ; ethnology ; Male ; Middle Aged ; Tibet ; Transients and Migrants ; Young Adult

Objective To evaluate the clinical application and significance of lengthen-stem bipolar-femur prosthetic replacement for the treatment of old-age patients with intertrochanteric fracture osteoporosis.Methods 28 cases of patients aging from 75 to 99 years old of intertrochanteric fracture osteoporosis treated with lengthen- stem cemented bipolar prosthesis were studicd from March 2000 to December 2006.After taking the blank margin, the bones of different sizes were replaced and the steel wire was fixed.After determining the depth of the front an- gle,the artificial bone was placed.Results After 28 examples attaining the following-up examination for 7 months to 3 years,with an average of 1.5 years,its function according to Harris standard was evaluated 3 months after the operation,8 examples were excellent,13 examples good,5 examples pass,2 examples inferior.The excellent or good rate reached 75% ,with no abnormal cases,no joint dislocation during the followed-up period.1 example had the phantom phenomenon 1 year after the operation.2 examples among the inferior had got more serious internal medicine disease which affected the restoring function.1 example died of the internal medicine disease 1 year after the operation.Conclusion By using the lengthen-stem bipolar-femur prosthetic replacement for the treatment of old patients with inrertrochanteric fracture osteoporosis,the patients will restore quickly after the operation and can carry a heavy load at an early time.The illness complication and the mortality rate will be redaced.But its related disease must be strictly dealt with and the surgery operating skill must be grasped.

OBJECTIVETo explore the application of Dried Blood Spot (DBS) testing for early detection of HIV infection among infants.

METHODSAll of the infants aged between 6 weeks and 18 months and born by HIV positive mothers from 14 Maternity and Child Health Care Hospitals in Kunming, Dali, Dehong, Lincang of Yunnan province were investigated from 2010 to 2011. By using DBS and Roche HIV-1 DNA test techniques, 286 infants were tested for HIV early diagnosis and compared with HIV antibody results of 18 months infants. DBS from uninfected infants were taken periodically and screened of HIV antibody to find their time of antibody-disappearing. The information of treatment for pregnant women and feeding methods for infants was also investigated.

RESULTSA total of 286 infants were tested with HIV-1 DNA among which 148 infants were male and 138 infants female, and 8 infants were HIV-1 DNA positive and the infection rate was 2.8% (8/286) that was in accord with their antibodies results in 18 months old; the other 278 infants whose HIV-1 DNA was negative was also negative with their antibodies. By following up the antibody test of 143 HIV negative infants the cumulate rates of antibody-disappearing at the age of 6, 9, 12 and 18 months were 14.0% (20/143), 61.5% (88/143), 88.1% (126/143) and 100.0% (143/143), respectively. Among 286 HIV positive pregnant women, the group with anti-viral treatment had a lower rate of HIV infection with their infants that was 2.14% (6/280) while the group without anti-viral treatment had a high rate of HIV infection with their infants that was 33.33% (2/6). There was significantly different in the rates of two groups (P < 0.01). The HIV infection rate of infants fed with milk powder was 2.55% (7/274) and the rate was 8.33% (1/12) with breast milk.

CONCLUSIONThe HIV-1 DNA detection techniques with DBS sample was effective for the early diagnosis of HIV in infants from 6 weeks to 18 months.

DNA, Viral ; blood ; Dried Blood Spot Testing ; Early Diagnosis ; Female ; HIV Infections ; diagnosis ; HIV-1 ; genetics ; Humans ; Infant ; Infectious Disease Transmission, Vertical ; prevention & control ; Male

OBJECTIVETo assess the feasibility of visual-reality technique for simulating surgical resection of pancreatic tail carcinoma using a 3-dimensional pancreas model reconstructed on the basis of the CT data.

METHODSThe original image data of 64-slice spiral CT was obtained from a patient with pancreatic tail carcinoma. Using adaptive region growing algorithm, the serial CT images were segmented and automatically extracted for 3-dimensional reconstruction of the pancreas and the anatomically related structures with a self-designed program. The model was then processed with Freeform Modeling System for image modification and smoothing. With the assistance of GHST SDK and PHANTOM software systems, preoperative simulation of surgical resection of the carcinoma was performed on the basis of the established pancreatic model.

RESULTSThe reconstructed 3-dimensional pancreatic model with the related structures clearly visualized the 3-dimensional structures of the pancreas, the pancreatic tail compromised by the carcinoma, and the adjacent organs, displaying also the distribution, courses and the anatomical relations of the ductal systems including the main pancreatic duct, abdominal aorta, portal vein system, and the biliary tract. During simulated surgery for pancreatic tail carcinoma resection, the GHOST SDK system allowed effective application of the virtual surgical instruments, and the use of PHANTOM software produced a surgical experience with high resemblance of that from an actual operation.

CONCLUSIONThe serial CT data-based reconstruction of 3-dimensional pancreas model and simulated operation on this model using virtual-reality technique has great potentials for application in individualized surgical planning and surgical risk assessment in cases of pancreatic tail carcinoma, and also facilitates clinical training of the surgeons.

Computer Simulation ; Female ; Humans ; Imaging, Three-Dimensional ; methods ; Middle Aged ; Models, Biological ; Pancreatic Neoplasms ; diagnostic imaging ; surgery ; Tomography, Spiral Computed ; User-Computer Interface

OBJECTIVETo study the two-dimensional (2D) image segmentation, three-dimensional (3D) reconstruction and virtual surgery of cholecystectomy based on the 2D image data of the liver, biliary track and cholecystolithiasis obtained by 64-slice spiral CT.

METHODSThe image data of the liver, biliary track and cholecystolithiasis were obtained by 64-slice spiral CT scanning. Segmentation and automatic extraction of the images were performed using auto-adapting region growing algorithm. 3D reconstruction of the segmented data was carried out using MIMICS10.0 and self-designed software, and the data of the 3D model of the liver with the billiary tract were imported into FreeForm Modeling System for registration and smoothing. Virtual surgery of cholecystotomy for calculus removal and cholecystectomy were performed with Phantom.

RESULTSThe auto-adapting region growing algorithm allowed rapid image segmentation, and the 3D model of the liver based on the segmentation data clearly displayed vivid 3D structures of the liver. Virtual operations of cholecystectomy could be performed in the FreeForm Modeling System.

CONCLUSIONThe algorithm we proposed can correctly and rapidly complete image segmentation and 3D reconstruction of cholecystolithiasis from the data 64-slice spiral CT, and allows virtual operations on the gallbladder.

Cholecystectomy ; Cholecystolithiasis ; diagnostic imaging ; surgery ; Cholecystostomy ; Computer Simulation ; Computer-Assisted Instruction ; methods ; Humans ; Imaging, Three-Dimensional ; methods ; Surgery, Computer-Assisted ; methods ; Tomography, Spiral Computed ; User-Computer Interface

OBJECTIVETo detect the expression and variation of p53 gene during tree shrews' hepatocarcinogenesis induced by hepatitis B virus (HBV) and aflatoxin B1 (AFB1).

METHODSTree shrews were divided into four groups: the tree shrews were infected with HBV and fed with AFB1 in group A, only infected with HBV in group B, fed with AFB1 alone in group C, and normal control in group D. All the tree shrews were performed liver biopsy every 15 weeks. The tissues of liver and tumor were detected by immunohistochemistry and molecular biotechnologies.

RESULTS(1) The incidence of hepatocellular carcinoma (HCC) in group A (66.7%) was higher than that in Group B and C (30%). HCC appearance in group A was earlier than that in group C (120.0 weeks +/-16.6 weeks vs 153.3 weeks +/-5.8 weeks, t = 3.336, P<0.01). (2) Mutated p53 protein was not found before the 75th week of the experiment in each group. (3) At the 105th week, the expression rates of mutated p53 protein were 78.6%, 60% and 71.4% in group A, B and C respectively, which were much higher than that (10%) in group D (x2 > or = 5.03, P<0.05). An abnormal band of p53 gene was detected in both group A and C. (4) The mutation points of p53 gene in liver cancer of tree shrew were at codon 275, 78 and 13. The nucleotide sequence and amino acids sequence of tree shrew's wild-type p53 showed 91.7% and 93.4% homology with those of human p53 respectively.

CONCLUSIONSThere is a remarkable synergistic effect between HBV and AFB1 on HCC. Mutated p53 protein is expressed before HCC occurrence, which promotes the development and progress of HCC. HBV and AFB1 may synergistically induce p53 gene mutation.

Aflatoxin B1 ; toxicity ; Animals ; Carcinoma, Hepatocellular ; genetics ; Cocarcinogenesis ; Gene Expression Regulation, Neoplastic ; Genetic Variation ; Hepatitis B ; virology ; Hepatitis B virus ; Liver Neoplasms, Experimental ; genetics ; Point Mutation ; RNA, Neoplasm ; analysis ; Tumor Suppressor Protein p53 ; genetics ; Tupaiidae