Objective To compare the differences of clinical characteristics between genotype B and C chronic hepatitis B(CHB)patients and to summarize clinical factors related to genotype C hepa- titis B virus(HBV)infection.Methods Seventy eight CHB patients who were diagnosed with genotype B or C infection by liver puncture biopsy and genotyping were enrolled.Their serum HBV DNA levels were detected.Severe hepatitis,liver cirrhosis,hepatocellular carcinoma and HBeAg positive rate were analyzed to determine the pathologic inflammation and fibrosis degree of liver tissue.Chi square test and Logistic multiple regression analysis were employed for the statistical analysis.Results The serum albumin and pre-protein were lower in genotype C CHB patients than that in genotype B.The alanine aminotrans- ferase,total bilirubin and prothrombin time were higher in genotype C CHB patients than that in genotype B.The rates of genotype C patients increased significantly with the grade of liver necroin- flammation progressing from GO to G4(1.8%,11.1%,20.4%,33.3%,33.3%) and the stage of liver fibrosis progressing from SO to S4(5.6%,5.6%,14.8%,33.3%,40.7%),but the rates of genotype B patients did not change significantly with the grade of liver necroinflammation(16.7%, 25.0%,25.0%,20.8%,12.5%)and stage of liver fibrosis progressing(16.7%,29.2%%,20.8%, 16.7%,16.7%).There was statistical significance in grades of liver necroinflammation(X~2= 11.49,P=0.022)and stages of liver fibrosis(X~2=13.56,P=0.006)between genotype B and gen- otype C patients.The rates of genotype C CHB patients were higher than,similar with and lower than the rates of genotype B patients of HBV DNA level above 1.0?10~6 copy/mL,between 5.0?10~2-1.0?10~6 copy/mL and under 5.0?10~2 copy/mL,respectively(51.8% vs 12.5%,35.2% vs 45.8% and 13.0% vs 41.7%).There was statistical significance of HBV loads between genotype B and genotype C patients(X~2=13.25,P=0.001).HBeAg positive rate in genotype C patients was significantly higher than that in genotype B patients(61.1% vs 25.0%,X~2=8.67,P=0.003).The rates of decompensated cirrhosis,compensated cirrhosis and no-cirrhosis in genotype C patients were higher than,similiar with and lower than the rates in genotype B patients,respectively(40.7% vs 4.2%,22.2% vs 20.8% and 37.0% vs 75.0%).There was statistical significance of the rate of cirrhosis between genotype B and genotype C patients (X~2=12.47,P=0.002).Conclusions The degree of liver necroinflammation and fibrosis,the HBeAg positive rate and the incidence of cirrhosis are all related with genotype C HBV infection.

Antiviral Agents ; adverse effects ; therapeutic use ; Female ; Hepatitis C, Chronic ; complications ; drug therapy ; psychology ; Humans ; Interferon-alpha ; adverse effects ; therapeutic use ; Mental Disorders ; complications ; Middle Aged

OBJECTIVETraditional prenatal diagnosis for congenital diseases were villus sampling and amniocentesis. These invasive diagnosis methods are not only technical complicated, but also harmful to mother or fetus. Fetus in its different gestational age has its different type of hemoglobin or different amount of hemoglobin, especially epsilon hemoglobin exiting in the body of 10 weeks gestation fetal, however gamma hemoglobin has its high amount before baby to be born. But epsilon and gamma hemoglobin did not exist in the bodies of adults bodies. It is possible to use advanced molecular biological technique to extract the fetal hemoglobin gene from maternal peripheral blood. In articles from domestic and abroad, no report related to fetal hemoglobin extraction from maternal peripheral blood was found. We tried to use non-invasive method to detect fetal hemoglobin epsilon/gamma gene from maternal peripheral blood by molecular biological technique. The purpose was to establish a convenient, sensitive and special method to be a basis of screening prenatal diseases in the population and lay a basis for family planning and clinical application.

METHODSBlood samples were collected and the fetal mRNA extracted from the pregnant women with the use of random primer. We used ultraviolet spectrophotometer to test the concentration and purity of extracted mRNA are suitable for reverse transcription. Reverse transcription of mRNA into cDNA was carried out and cDNA by PCR with the special epsilon/gamma primer being used. Via 1.2% EB in agarose gel electrophoresis, we used "Gel Works System" to scan the electrophoresis image to detect epsilon/gamma gene band.

RESULTSThe peripheral blood of pregnant women was collected. With RT-PCR and agarose gel electrophoresis method, we detected epsilon/gamma gene successfully in 7 samples with 6 positive and 1 negative.

CONCLUSIONThis was the first time that we used non-invasive way to detect expression of fetal epsilon/gamma gene in maternal blood to have found that this was a simple method to separate fetal cells from maternal blood, and could easily be accepted by pregnant women. Success of RT-PCR to detect fetal specific mRNA gave the hint that this method could be used in the field of prenatal diagnosis of hemoglobin disease, predicting fetal gender, predicting Rh blood type and single gene disease and be used widespread in prenatal diagnosis.

Female ; Globins ; genetics ; Humans ; Pregnancy ; blood ; RNA, Messenger ; blood ; Reverse Transcriptase Polymerase Chain Reaction ; methods

Stirred culture offers a number of advantages over static systems as it maintains a stable, homogeneous culture environment and is easy to scale-up. This paper focused on the development and application of stirred tank bioreactor to culture hematopoietic cells. Preliminary study of stirred culture of hematopoietic cells was carried out in cord blood mononuclear cells culture in spinner flask. The results showed that the amplification rates of total cell, CFU-GM and BFU-E, with the exception of CFU-Mk, were greater in spinner flask than T-flask. The number of total cells increased 20 fold after 14 days incubation in spinner flask. The amplification rates of CFU-GM, CFU-Mk and BFU-E reached maximum at 10th day, 10th day and 7th day respectively, and the maximal amplification rates were 9.2-fold, 5.5-fold and 2.4-fold respectively, whereas the rate of CD34+ cells in spinner flask was (6.7 +/- 4.0)-fold at day 10. These results indicated that the stirred culture system is better than the static culture systems for hematopoietic cell proliferation. The biocompatibility of cord blood MNC to different types of materials used in bioreactors was also tested. The results showed that glass, stainless steel 316L and polytetraflouroethylene (PTFE) supported the growth of hematopoietic cells well. A higher cell density was reached in stirred bioreactors with controlled pH and DO than static culture. These findings suggested that the controlled large-scale culture could be used to overcome the clinical shortage of hematopoietic cells.
Antigens, CD34 ; metabolism ; Bioreactors ; Cell Culture Techniques ; instrumentation ; methods ; Erythroid Precursor Cells ; cytology ; Fetal Blood ; cytology ; Granulocyte-Macrophage Progenitor Cells ; cytology ; Humans ; Polytetrafluoroethylene ; Stainless Steel

OBJECTIVETo construct and express human CD96 gene outer membrane domain (hCD96om) in prokaryotic cells and prepare rabbit polyclonal antibody of hCD96om.

METHODShCD96om was amplified by RT-PCR from the peripheral blood of patients with acute myeloid leukemia and inserted into prokaryotic expression vector pET32a(+) to construct the recombinant plasmid pET32-CD96. The expression of hCD96om was induced by IPTG in BL21(DE3) cells, and the expression product was identified by Western blotting. The anti-hCD96 polyclonal antibody was prepared by immunization of rabbits with the fusion protein. The specificity of anti-hCD96 antibody was determined by Western blotting.

RESULTShCD96om protein was expressed in E.coli BL21(DE3) cells in the form of inclusion body, with a relative molecular mass around 37 kD. Western blotting showed a specific reaction of the prepared antiserum with the 70 kD protein extracted from human leukemia cell line HL-60 cells and with the 37 kD hCD96om fusion protein.

CONCLUSIONThe CD96 gene of human has been successfully cloned and expressed in BL21(DE3) cells, and its rabbit polyclonal antibody has been obtained.

Animals ; Antibodies ; immunology ; metabolism ; Antigens, CD ; biosynthesis ; genetics ; immunology ; Base Sequence ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Humans ; Immune Sera ; biosynthesis ; Immunization ; Leukemia, Myeloid, Acute ; immunology ; Molecular Sequence Data ; Neoplastic Stem Cells ; immunology ; Rabbits ; Recombinant Proteins ; biosynthesis ; genetics ; immunology

OBJECTIVETo develop a coated electrode of immobilized denitrificants and to evaluate the performance of a bioelectrochemical reactor to enhance and control denitrification.

METHODSDenitrifying bacteria were developed by batch incubation and immobilized with polyvinyl alcohol (PVA) on the surface of activated carbon fiber (ACF) to make a coated electrode. Then the coated electrode (cathode) and graphite electrode (anode) were transferred to the reactor to reduce nitrate.

RESULTSAfter acclimated to the mixtrophic and autotrophic denitrification stages, the denitrifying bacteria could use hydrogen as an electron donor to reduce nitrate. When the initial nitrate concentration was 30.2 mg NO3- -N / L, the denitrification efficiency was 57.3% at an applied electric current of 15 mA and a hydraulic retention time (HRT) of 12 hours. Correspondingly, the current density was 0.083 mA/cm2. The nitrate removal rate of the reactor was 34.4 g NO3- -N/m3 x d, and the surface area loading was 1.34 g NO3- -N / m2 x d.

CONCLUSIONThe coated electrode may keep high quantity of biomass, thus achieving a high denitrification rate. Denitrification efficiencies are related to HRT, current density, oxidation reduction potential (ORP), dissolved oxygen (DO), pH value, and temperature.

Adsorption ; Bacteria ; metabolism ; Biodegradation, Environmental ; Bioreactors ; Carbon ; chemistry ; Electrodes ; Electrolysis ; Nitrates ; metabolism ; Oxidation-Reduction ; Oxygen ; Polyvinyl Alcohol ; Temperature ; Time Factors ; Water Pollutants, Chemical ; metabolism ; Water Purification ; methods

Objective To build the dataset of Chinese visible human female. Methods After undergoing macroscopical, CT and MRI examinations to exclude organic lesions, a young female cadaver of medium height was selected as the subject. After morphological measurement and vascular perfusion, the cadaver was embedded with 5% gelatin and cryopreserved in a -30 ℃ icehouse for 1 week. A digital milling machine TK 6350 (milling accuracy of 0.001 mm) was used to shave off slices of the body layer by layer from head to foot in a laboratory at -25 ℃. The successive cross sections were photographed with a high definition digital camera, and the pictures were put into a computer to establish a dataset of human body. By utilizing the image dataset derived from the successive cross sections, 3D reconstruction and stereodisplay of human structure were finished with a SGI Workstation which was equipped with an independently self developed software package for 3D reconstruction. Results The selected specimen, a 22 year old female native of Chongqing, was 1 620 mm in height, 54 kg in weight and died of non organic disease. CT scans were made in every 1.0 mm for head and neck and every 2.0 mm for rest parts, and the thickness for MRI scans was 1.5 mm for head and 3.0 mm for rest parts. For serial cross sections, the thickness was 0.25 mm for head and 0.5 mm for rest parts. Thus, a total of 3640 slices were obtained, and the photo for every slice was saved as a 36 MB file in a resolution of 6 291 456 pixels (3 072?2 048). Finally, the complete data files reached to 131.04 GB. Conclusion ① This is the first formally reported case of Chinese visible human female, suggesting that China becomes the second country owning visible human female dataset of her population. We set up a website for the purpose of exchanging ideas and information on this subject. So, the results are issued simultaneously on the Internet (http://www.chinesevisiblehuman.com).② According to US Visible Human Project(VHP), the data of the 3 junctional parts of their female cadaver were absent because the body was cut into 4 segments. Taking the age of 59 year old into account, the visible human female's body was not exactly perfect. The sections of 0.33 mm in thickness were saved to pictures at a resolution of 2 490 368 pixels (2 048?1 216). While, the first Chinese visible human female reported here is a young female without organic disease or lesion. No sectional datum is lost for being acquired from successive sections of the whole body. The resolution of cross sectional image reaches to 6 291 456 pixels (3 072?2 048).

Objective To achieve computer visualization of the first Chinese visible male and female Methods After acquisition of the dataset of the first Chinese visible male and female (2 518 cross sections were obtained from the visible male, the complete data files take up 90 468 GBs; while 3 640 cross sections from the female, the complete data files take up 131 04 GBs ), we processed 2 D images in an SGI Workstation and on P4 computer respectively Then, image registration was performed through reserved scaling point Reconstruction was achieved by two approaches: volume rendering reconstruction and surface rendering reconstruction Results We visualized the whole body and special parts of Chinese visible male and female on an SGI Workstation and a personal computer respectively Furthermore, by optimizing 3 D reconstruction and data processing technique, interactive 3 D visualization of the dataset was achieved Conclusions ①The dataset of the first Chinese visible male and female proves to be eligible for 3 D visualization research ②The platform setup of interactive 3 D visualization of Chinese visible male and female dataset provides foundation for digital human anatomy and virtual surgery ③The models of human organs and parts built through data segmentation, classification, registration and drawing lay basis for rendering complex structures of the whole human body delicately

Objective To establish more detailed dataset of Chinese visible human male. Methods After undergoing macroscopical, CT and MRI examinations to exclude organic lesion, a young aged, middle sized male cadaver was selected as the subject. First, morphological measurement and vascular perfusion were performed. Second, after embedding with 5% gelatin, the cadaver was put in ice house and frozen to -30 ℃ for 1 week. Third, TK 6350 numerical control milling machine (milling accuracy of 0.001 mm) was used to shave off slices of the body layer by layer from head to foot at -25 ℃ in low temperature laboratory. Fourth, the successive cross sections were photographed with high resolution digital camera and scanned into an animation computer. Thus, data acquisition from cadaver model was completed to obtain structural dataset of the human body. Results The selected sample was a 21 year old, 1 820 mm in height, 66 kg in weight male died due to non organic disease. CT with 1.0 mm slice thickness for the head and neck and 2.0 mm for the rest of the body was performed. MRI with 1.5 mm slice thickness for the head and neck and 3.0 mm for the rest of the body was also performed. A total of 18 398 serial cross sections with the thickness of 0.1 mm of each section were obtained. The digital photographs were sampled at a resolution of 10 989 056 (4 064?2 704) pixels. The data file of each section occupies 62.9 MB. The complete data files occupy 1 157.23 GB. The research results are issued simultaneously on the Internet (http://www.chinese visiblehuman.Conclusion ① Review of the related literatures reveals that the thinnest thickness of the reported cross section of the visible human dataset is 0.2 mm(the thickness of the sections of the skull base of the first case of Chinese visible human reported by our research group is 0.1 mm.), and the slices consist of several thousands of serial cross sections with several millions of pixels. The data files occupy several tens of GB or more than 100 GB. However, the thickness of the cross sections of the whole body of the dataset achieved in our research is 0.1 mm. The total slices consist of 18 398 serial cross sections with the photographic resolution of 11 million pixels and the total data file reaches 1 157.23 GB. The three indexes mentioned above are elevated by 1 log unit. ② We have solved the key technical problems in data acquisition of visible human such as super thin serial cross sectioning, enormous quantity of data storing and display of tiny blood vessels.