Extraction of protoporphyrin disodium and its inhibitory effects on HBV-DNA

AIM:To explore an ideal method for extracting protoporphyrin disodium (PPN) from unanticoagulated animal blood, and to study the inhibitory effects of PPN on HBV-DNA duplication and its cytotoxicity to 2.2.15 cell strain.METHODS:Protoporphyrin methyl ester and other intermediate products were prepared with protoheme separated from protein hydrolysates of coagulated animal blood, which were finally made into PPN and detected quantitatively with an ultraviolet fluorescent analyzer.Ten μg/ml, 20μg/ml, 40μg/ml, 80μg/ml and 160μg/ml of PPN-aqueous solution were added into culture medium for 2.2.15 cells respectively. Eight days later, the drug concentration in supernatant from the culture medium was detected when inhibition rate of HBeAg, cell survival rate when inhibition rate of HBeAg was 50% (ID50), and when survival cells in experimental group were 50% of those in control group (CD50), and the therapeutic index (TI) was also detected. PPN with different concentration of 10μg/ml,20μg/ml, 40μg/ml, 80μg/ml and 160μg/ml was respectively mixed and cultivated with HepG2 2.2.15 cell suspension,and then the inhibition of PPN against HBV-DNA was judged by PCR.RESULTS:The extract of henna crystal was identified to be PPN. When the concentrations of PPN were 160μg/ml and 80μg/ml, the inhibition rates of HBeAg were 89.8% and 82.4%, and the cell survival rates were 98.7% and 99.2%.CONCLUSION:It is suggested that PPN can be extracted from unanticoagulated animal blood. PPN can inhibit HBV-DNA expression and duplication in vitro, and has no cytotoxicity to liver cells. Further study and application of PPN are warranted.

Clinical and epidemiological features of patients with clonorchiasis

AlM:To study the clinical and epidemiological features of patients with clonorchiasis so as to provide scientific evidences for the diagnosis and prevention of clonorchiasis.METHODS:Stools from 282 subjects suspected of having clonorchiasis were examined for helminth eggs with modified Kato's thick smear and sedimentation methods,and their sera were tested for HAV-DNA, HBV-DNA, HCV-RNA, HDV-RNA and HEV-RNA with polymerase chain reaction (PCR).Clinical symptoms of patients with clonorchiasis only were analyzed, and their blood samples were tested for circulating antigen (CAg) with Dot-ELISA, esoinophilic granuloolte count,and alanine aminotransferase (ALT). Meanwhile, they were asked to provide data of occupation, eating habit, hygienic habit and knowledge of donorchiasis. In addition, the ecosystem of the environment in epidemic areas was surveyed.RESULTS:Among the 282 patients,61 (21.43%) were infected with clonorchis sinensisonly,97 (34.64%) were co-infected with clonorchis sinensis and other pathogens,92 (32.86%) were infected with hepatitis virus only and 31 (11.07%) neither with clonorchis sinensis nor hepatitis virus.Among the 61 patients with clonorchiasis only, there were 14 (22.95%) subjects with discomfort over hepatic region or epigasfrium,12 (19.67%) with general malaise or discomfort and inertia in total body, 6 (9.84%) with anorexia, indigestion and nausea,4 (6.56%) with fever, dizziness and headache (6.56%), and 25 (40.98%) without any symptoms, sixty one (100%) with CAg(+), 98.33% (59/60) with eosinophilic granulocytes increased and 65.00% (39/60) with ALT increased. B-mode ultrasonography revealed 61 cases with dilated and thickened walls of intrahepatic bile duct, and blurred patchy echo acoustic image in liver. Twenty-six cases had stones in the bile duct, 39 cases had slightly enlarged liver with diffuse coarse spots in liver parenchyma. Twenty cases had enlarged gallbladder with thickened coarse wall and image of floating plagues, 9 cases had slightly enlarged spleen. By analysis of epidemiological data, we found that the ecologic environment was favorable for the epidemiology of clonorchiasis.Most patients with clonorchiasis were lack of knowledge about the disease.Their living environment,hygienic habits, eating habits and their occupations were the related factors that caused the prevalence of the disease.CONCLUSION:The clinical symptoms of clonorchiasis are non-specific, and the main evidences for diagnosis of clonorchiasis should be provided by etiologic examination,B-mode ultrasonography and clinical history.The infection of clonorchis sinensis is related to occupations, bad eating habits and lack of knowledge about prevention of the disease.

Effect of alpha 2b interferon on inducement of mIL-2R and treatment of HCV in PBMC from patients with chronic viral hepatitis C

AIM: To study the level of membrane interleukin-2 receptor (mIL-2R) on surface of peripheral blood mononuclear cells (PBMC) and the therapeutic efficacy of alpha 2b interferon on the treatment of HCV-RNA in PBMC of patients with chronic hepatitis C and to compare the negative rates of HCV-RNA in PBMC, HCV-RNA and anti-HCV in serum.METHODS: Before and after treatment of alpha 2b interferon, the level of mIL-2R of patients with chronic hepatitis C was detected by biotin-streptavidin (BSA). The therapeutic group (26 cases) was treated with alpha 2b interferon (3 MU/d) and control therapeutic group (22 cases)was treated with routine drugs (VitC, aspartic acid). The total course of treatment with alpha 2b interferon and routine drug was six months and per course of the treatment was three months. The levels of HCV-RNA in PBMC, HCV-RNA and anti-HCV in serum were detected before and after a course of the treatment.RESULTS: Before and after treatment of alpha 2b interferon and routine drugs, the levels of mIL-2R in silence stage were (3.44±0.77)% and (2.95±0.72)%, the levels of mIL-2R in inducement stage were (33.62±3.95)% and (30.04±3.73)%. There was a significant difference between two groups (P＜0.01-P＜0.05). After treatment of alpha 2b interferon with 3 MU/d for two courses of the treatment,the total negative rates of HCV-RNA in the PBMC and HCVRNA, anti-HCV in serum were 42.31% (11/26), 57.69%(15/26), 65.38%(17/26) respectively. After the treatment of routine drug, the negative rates of HCV-RNA in PBMC and HCV-RNA, anti-HCV in serum were 13.64% (3/22),22.73% (5/22), 27.27% (6/22) respectively. There was high significant difference in the group treated with alpha 2b interferon and the group treated with routine drugs (P＜0.01-P＜0.05).CONCLUSION: The mIL-2R can be induced by alpha 2b interferon during the treatment. The alpha 2b interferon has a definite effect on the treatment of HCV-RNA in PBMC.The curative effect of alpha 2b interferon is better than that of the routine drugs.

Study on the Cellular Molecular Mechanism of Intrauterine Transmission of Hepatitis B Virus

Objective: To study intrauterine transmission of HBV and its cellular molecular mechanism and influence on the fetus. Methods.. A total of 46 coses of pregnant women who suffered from HBV were divided into HBeAg ( + ) and HBeAg ( -) groups. HBV-DNA in serum and peripheral blood mononuclear cells (PBMC-) of 46 cases of pregnant women before delivery was detected by polymerase chain reaction (PCR). After placenta being delivery, HBV-DNA in serum and cord blood mononuclear cells (CBMC) was also detected by PCR. Results.. The total of positive rates of HBV-DNA in serum and PBMC of pregnant women with hepatitis B were 69. 57% (32/46) and 41. 30% (19/46). The positive rates of HBV-DNA in serum of cord blood and CBMC were 56. 52%(26/46) and 21. 74% (10/46) respectively. Among them, the positive rates of HBV-DNA in serum and PBMC of pregnant women with HBeAg ( + ) were 100. 00% (25/25) and 60. 00% (15/25) respectively. The positive rates of HBV-DNA in serum of cord blood and CBMC were 88. 00% (22/25) and 32. 00% (8/25) respectively. The positive rates of HBV-DNA in serum and PBMC of pregnant women with HBeAg (-) were 33. 33% (7/21) and 19.05% (4/21) respectively. The positive rates of HBV-DNA in serum of cord blood and CBMC were 19. 05%(4/21) and 9. 52% (2/21) respectively. The positive rates of HBV-DNA in serum of cord blood and CBMC of newborns were higher in the group of pregnant women with HBeAg ( + ) than those in the group of pregnant women with HBeAg ( -) (P<0. 01 and P<0. 05). There was no HBV-DNA in serum, PBMC and CBMC of normal pregnant women and normal neonates. Conclusion: The intrauterine transmission of HBV can be existent and its transmission way not only can be induced by serum but also can be induced by PBMC. The way of intrauterine transmission of HBV induced by PBMC was concealed. The dangerous possibility of intrauterine transmission is higher in the pregnant women with HBeAg (+) than that in the group of pregnant women with HBeAg ( -).

Similar Single-Difference Model and Its Algorithm for Solving GPS Monitoring Deformation Directly at Single Epoch

A new similar single-difference mathematical model (SS-DM) and its corresponding algorithmare advanced to solve the deformationof monitoring point directly in singleepoch. The method for building theSSDM is introduced in detail, and themain error sources affecting the accu-racy of deformation measurement areanalyzed briefly, and the basic algo-rithm and steps of solving the deform-ation are discussed.In order to validate the correctnessand the accuracy of the similar single-difference model, the test with fivedual frequency receivers is carried outon a slideway which moved in plane inFeb. 2001. In the test,five sessions areobserved. The numerical results oftest data show that the advanced mod-el is correct.

Angular distribution of sputtered atoms induced by low-energy heavy ion bombardment

The sputtering yield angular distributions have been calculated based on the ion energy dependence of to- tal sputtering yields for Ni and Mo targets bombarded by low-energy Hg+ ion. The calculated curves show excellent agreement with the corresponding Wehner’s experimental results of sputtering yield angular distribution. The fact clearly demonstrated the intrinsic relation between the ion energy dependence of total sputtering yields and the sput- tering yield angular distribution. This intrinsic relation had been ignored in Yamamura’s papers (1981,1982) due to some obvious mistakes.

Diagnosis of intestinal acariasis with avidin-biotin system enzyme-linked immunosorbent assay

AIM: To explore the value of avidin-biotin system enzyme-linked immunosorbent assay (ABC-ELISA) in diagnosis of intestinal acariasis.METHODS: Mite-specific IgG levels in serum of 48 patients with intestinal acariasis were measured with ABC-ELISA.The sensitivity of this method was compared with that of staphylococcal protein A enzyme-linked immunosorbent assay (SPA-ELISA).RESULTS: The positive rate of mite-specific IgG detected with ABC-ELISA and SPA-ELISA was 89.58% (43/48) and 56.25% (27/48), respectively. The positive rate with ABC-ELISA was statistically higher than that with SPA-ELISA (x^2= 13.50, P<0.01).CONCLUSION: ABC-ELISA is an effective method for the diagnosis of intestinal acariasis.

Cloning and Subcloning of cDNA Coding for Group Ⅱ Allergen of Dermatophagoides Farinae

Objective: To clone, sequence and subclone the cDNA coding for group Ⅱ allergen of Dermatophagoidesfarinae(Der f2). Methods: The cDNA gene fragment of Der f2 was amplified by RT-PCR. After being purified, the gene fragment was cloned into a vector pMD-18T. The recombinant plasmid pMD-18T-Der f2 was transformed into E. coli JM109. Positive clones were screened and identified by PCR and digested with restriction endonuclease, and the sequence of inserted Der f2 cDNA was also analyzed. Then Der f2 was subcloned into the vector of pET-32a(+). Results: The Der f2 cDNA was specifically amplified from RNA by RTPCR. The recombinant plasmid pMD-18T-Der f2 and pET-32a(+),Der f2 was constructed and digested by SacⅠ and NotⅠ, and the size of gene fragment was 455bp and in accordance with the expected one. Conclusion: The pET-32a(+)-Der f2 subclone was constructed successfully.

Hamilton Graph Based on DNA Computing

DNA computing is a novel method for solving a class of intractable computationalproblems in which the computing can grow exponentially with problem size. Up to now, manyaccomplishments have been achieved to improve its performance and increase its reliability.Hamilton Graph Problem has been solved by means of molecular biology techniques. A smallgraph was encoded in molecules of DNA, and the “operations” of the computation wereperformed with standard protocols and enzymes. This work represents further evidence forthe ability of DNA computing to solve NP-complete search problems.

Geochemistry of Sulfur and Hazardous Elements in Late Paleozoic Coals and Partings from Taozao Coalfield, Shandong Province, China

The analyses of different sulfur forms, the trace elements in pyrites using electron microprobe and the trace elements in coal using INAA (instrumental neutron activation analysis) of the Late Paleozoic coals from the Taozao coalfield in Shandong Province, China, conclude that most sulfur (>75 %) in high-sulfur coal of Taiyuan Formation occurred as pyrite, in which many hazardous elements co-existed and their concentrations varied with their geological origin. The concentrations of hazardous elements in high-sulfur coals from Taiyuan Formation, composing mainly of Cu, As, U, Pb, Mo and Co, are much higher than those in the low-sulfur coals from Shanxi Formation and Shihezi Formation, because the influence of seawater during and after coal accumulation in Taiyuan Formation is stronger than those in Shanxi and Shihezi formations. Moreover, the element As is related to Fe, and both elements exist mainly in the form of pyrite. The element U is richer in the coal influenced by seawater. In addition, the coal affected by the magmatism contains more U, too. When high-sulfur coals are processed with heavy media washing to remove sulfur and minerals, the majority of hazardous elements will also be removed from the coals.

Application of Fuzzy-PID Control System in Full-Mechanized Coal Face

The control system, which includes structure, the composition of software and hardware, the form of PID control system and its systematic closed-loop, was used in No.4236 full-mechanized coal face of Xinlongzhuang mine. The typical fuzzy PID control system structure was investigated, and a simplified fuzzy PID control system was taken the place of the complex three-dimension fuzzy controller. Based on the parameter relation between fuzzy controller and normal PID controller, a common method of parameter adjustment of PID controller was summed up and the computer simulation was realized. This system can overcome the problems of large delay, nonlinear, poor running en- vironment and great load change in the full-mechanized coal face. The simulating investigation indicates that the de- signing method of fuzzy controller is simple and feasible.