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Viruses have been compatible with cells since the origin of life. The absolute dependence on cells leads to formation, development, distribution and evolution of viruses; the existence of viruses must have their long-term host for survival, spreading and extending. Like other species in nature, a group of viruses coexist with humans, which are called permanent resident virus in human being.The immune system of species has the function of anti-microbial invasion. The virus has a special coexistence mechanism with the long-term host. Because the immune system has no function to enter cells to recognize and remove viral nucleic acids, then invading viruses can be residual in cells, and as a recessive existence under the dynamic balance between residual viral replication and immune clearance pressure. This is the normal state of the virus with its long-term host coexisting harmoniously as stakeholders, and it also has been the normal state of all natural viruses after the immune system developed in host species, constituting the viral microecology of the host. The normal state viruses can undergo antigenic mutation to break through immune pressure and become abnormal, then viruses can proliferate again and spread to cause infection in host population, and would returned into normal state after a new immune protection to be established again. The primary infection of the normal state virus and the re-infection of the abnormal state virus can cause disease due to the immune toxicity induced by the immune response.The identification of permanent resident virus in human beings is of significant importance for studying the epidemiology, pathogenesis, clinical manifestations and outcomes of human viral diseases and epidemics, as well as prevention and treatment. Vaccine can successfully prevent the diseases caused by initial infection of normal state viruses. However, for the re-infection of abnormal state viruses, it is difficult to evaluate the preventive effect of vaccines because of the matching problem between previous vaccines and new antigens of abnormal state virus.
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Objective@#To explore the influence of pVR-IL15 gene adjuvant on the immune responses of different immunization strategies.@*Methods@#The sequential immunization strategies were used in BALB/c mice with a DNA vaccine, recombinant modified vaccinia virus Ankara and recombinant adenovirus expressing HBsAg respectively and combined with a gene adjuvant pVR-IL15. Cellular and humoral immune responses were evaluated by IFN-γ enzyme-linked immunospot assay (ELISPOT) and enzyme-linked immunosorbent assay (ELISA), respectively.@*Results@#The levels of humoral and cellular immune responses in the immune groups combined with pVR-IL15 were significantly higher than those of the non-combined with pVR-IL15.@*Conclusions@#The pVR-IL15 gene adjuvant can enhance the immune responses induced by the recombinant viruses expressing HBsAg.
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Objective To evaluate the anti-human respiratory syncytial viurs ( hRSV ) effect of aminoglucomannan ( AGM) in mice.Methods BALB/c mice infected with hRSV were randomly divided into AGM and konjac glucomannan ( KGM) groups with 54 in each.Then, mice in AGM group and KGM group were subdivided into three groups and treated with different doses of AGM or KGM as 2.5 μmol/L, 0.25 μmol/L or 0.025μmol/L.Each subgroup was further divided into 3 groups with 6 in each according to administration intervals of AGM or KGM as 8 h, 12 h or 24 h.All the mice are sacrificed at 72 h.The general condition of the mice was observed everyday.Reverse transcription-polymerase chain reaction ( RT-PCR) was used to detect the expression of hRSV fusion protein ( F) mRNA in the lung tissue of the mice. HE staining method and immunohistochemistry technique for intercellular cell adhesion molecule-1 ( ICAM-1) expression in the lung tissue were used to evaluate the inflammation in the lung tissue.ANOVA for randomized block design was used to examine the influence of dose and administration intervals on the expressions of hRSV F mRNA and ICAM-1.Results Reduced activity and asthma were observed in 16 mice in the KGM group, but not in the AGM group.And two mice in the KGM group died at d3, but there was no death in the AGM group.RT-PCR showed that the levels of hRSV F mRNA in lung tissues were 0.49 ±0.21 in AGM group and 0.88 ±0.06 in KGM group (t=6.71, P0.05).Conclusion AGM can alleviate the inflammation of lung tissue in mice infected with hRSV in a dose and time-depandent manner.
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Objective To explore the effect on traditional experiment and case teaching method in regional anatomy study. Methods 80 students from 2014 medical students were randomly selected as the teaching subjects and divided into traditional group and case teaching group. The traditional group con-tained 40 students, using the traditional teaching method, while case teaching group had also 40 students with case teaching method. In the process of teaching, three clinical cases were introduced, including thesubtotal thyroidectomy thoracic outlet syndrome andpancreatic cancer. After the end of the course, the students conducted a unified questionnaire and examination. SPSS 18.0 was used for data line t test or chi square test between the two groups. Results The scores of the students in the case group in the selection questions, blanks and essay questions in the final exam were higher than those of the traditional group; The average total score of the case group was (85.69 ±11.61), while the traditional group was (73.19 ±18.66), and the difference was statistically significant (t=3.597, P=0.002). The results of the questionnaire showed that the students in the case group were higher than the traditional group, and the difference was statistically significant ( χ2=14.753, P=0.001). Conclusion The effect on regional anatomy study with case teaching method is better than the traditional teaching method, and it is a promising teaching reform for the med-ical students.
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Objective To investigate the effects of ω-3 polyunsaturated fatty acids (ω-3 PUFA) on systemic inflammatory response and intestinal mucosa barrier in rats with traumatic brain injury and hemorrhagic shock (TBIS).Methods A total of 36 male Wistar rats were equally randomized into 3 groups:sham operation group,TBIS model group,and ω-3 PUFA pretreatment group.The serum levels of tumor necrosis factor-α (TNF-α),8-iso-prostaglandin F2 a (8-iso-PGF2 a),interleukin (IL)-1 β,and IL-10 were measured by enzyme linked immunosorbent assay (ELISA).HE staining was performed for morphological assessment of the intestinal tissue and evaluation of the intestinal mucosa damage index (IMDI).The marked bacilli of the mesenteric lymph nodes,lung,liver,spleen,and kidney tissue were counted under a fluorescent microscope.Results Compared with those in the sham group [ (38.15 ± 6.37) ng/ml,(84.91 ± 17.22) pg/ml,(2.52 ± 0.83 ) μg/L,(2.86 ± 0.82) μg/L,0.36 ±0.14,and 8.33％ ],the serum levels of TNF-α [ (328.11 ±20.09) and (244.37 ±21.82) ng/rrl],8-iso-PGF2a [ (263.47±55.19) and (176.35±41.63) pg/ml],IL-1β [ (27.06±2.61) and (18.91 ±1.78) μg/L],IL-10 [ (7.63 ± 1.29) and (9.52 ± 4.66) μg/L],the IMDI (4.18 ±0.39 and 3.31 ±0.40),and the positive rates of bacterial translocation (56.67％ and 35.00％ ) were significantly higher in both the TBIS model group and ω-3 PUFA group ( all P ＜ 0.01 ).Compared with TBIS model group,the levels of TNF-α,8-iso-PGF2 a,and IL-1 β,the IMDI,and the positive rate of bacterial translocation were significantly lower ( all P ＜ 0.05 ) and the levels of IL-10 were significantly higher in the ω-3 PUFA group (P ＜0.01 ).Conclusion The supplementation of ω-3 PUFA can remarkably inhibit the systemic inflammatory response and protect the integrity of intestinal mucosa in rat with TBIS.
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Objective To explore the distribution and clinical significance of hepatitis B core antigen( HBcAg) in the hepatocytes of chronic hepatitis B virus infected patients. Methods Paraffin sections were made from 41 liver biopsy samples obtained from chronic hepatitis B virus infected patients. The immuno-fluorescence confocal technique was utilized to analyze the expression level and localization of HBcAg in hepatocytes. The data were analyzed by using Kruskal Wallis test and chisquare test. Results HBcAg expression were detected in 36 (87. 8%) patients, among whom 23 cases had moderate abnormal liver function, 10 with mild abnormal liver function and 3 with normal liver function. Among the cases with moderate abnormal liver function, 6 cases showed the simple membrane-type HBcAg expression, 17 cases showed mixed cytosolic-type and membrane-type HBcAg expression without the nuclear-type expression. Twelve cases with mild abnormal liver function only showed simple cytosolic-type HBcAg expression without membrane-type or nuclear-type expression. In the three patients with normal liver function, HBcAg was expressed in cytoplasm and nuclear but not on membrane. The correlation between HBcAg expression pattern and liver function was statistically significant (χ2 =10. 60, P＜0.01). Conclusion HBcAg expression is directly correlated with liver injury in chronic hepatitis B virus infected patients, which indicates that membrane expressed HBcAg is the target antigen during the immuno-attack of liver.
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Objective To conduct item analysis on Work Attitude Scale(Wa) and to develop a short-versioned scale.Methods Wa and Symptom Checklist 90(SCL-90) were administered to 3767 recruits,item analysis of Wa was conducted and short version of Wa was developed based on the results of item analysis,Cronbach' s αcoefficient of the original scale and the short-versioned scale was calculated,criterion validity of the original scale and the short-versioned scale was analyzed using SCL-90 as the validity criterion.For other samples,16 Personality Factor Questionnaire (16PF),Eysenck Personality Questionnaire ( EPQ),and Self-Rating Anxiety Scale (SAS) were tested while the short-versioned scale was tested at the same time,criterion validity of the short-versioned Wa was analyzed using 16PF,EPQ,and SAS as the validity criterion.Results Difficulty of the original scale was from 0.03 to 0.87,27 items' difficulty was less than 0.20 or greater than.Discrimination index was from 0.04 to 0.56,22 items' discrimination index was less than 0.30.Correlation coefficient of the item score and the total score is from 0.02 to 0.52,18 items' correlation coefficient was less than 0.30.The short-versioned scale was made up of the 18 items with better quality.Cronbach' s α coefficient of the original scale and the short-versioned scale was 0.778 and 0.789,respectively.Both the total score of the original scale and the short-versioned scale were significantly correlated with the scores of SCL-90,and all the correlation coefficient were above 0.40.The correlation coefficient between the score of the short-versioned scale and that of the original scale was 0.934.The correlation coefficient between the score of the short-versioned scale and 16PF,EPQ,SAS was significant.Conclusion The short-versioned scale has a higher reliability and good validity.
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The HBV X gene was amplified by PCR according to the pecob6 containing the whole fragment of adw subtype of HBV, then the fragment was inserted into the multiple clone site of pAdTrack-CMV. The linearized shuttle plasmid was homogenously recombined with AdEasy-1 in BJ5183 and the recombinant adenoviral plasmid pAd-X was generated. Then plasmid pAd-X was digested with Pac I and transfected into 293 cells for packaging and amplifying. Infection titer and rate were monitored by green fluorescent protein (GFP) expression. With restriction endonuclease analysis and PCR methods, it has been confirmed that HBV X gene was cloned into the adenovirus vector successfully. The expression of X protein in HepG2 cells was detected by Western-blot. The recombined adenovirus Ad-X was constructed successfully, which would contribute to the advanced functional study of HBV X protein.
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Humans , Adenoviridae , Genetics , Metabolism , Genetic Vectors , Genetics , Metabolism , Green Fluorescent Proteins , Metabolism , Hep G2 Cells , Recombinant Proteins , Genetics , Trans-Activators , Genetics , TransfectionABSTRACT
Objective To study the condensation mechanism of aminoglucomannan(AGM)with plasmid DNA and some influencing factors in this process.Methods The polysaccharide-DNA complex was analyzed with agarose gel electrophoresis by observing the changes of DNA bands under various influencing conditions including different pH,ionic strength(NaCl concentration),temperature and plasmid DNA sorts.Results The new bands appeared in the slow migration positions after reaction between AGM and plasmid DNA,and the changes of DNA bands content were dependent on AGM concentrations.The condensation degree could be effected by pH,temperature,and ionic strength.Conclusion Our study suggests that the AGM can be condensed with plasmid DNA in proper conditions.
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Objective To observe the effect of rutin on growth and proliferation of human hepatic cancer line(HepG2).Methods HepG2 cells were cultured in vitro,then cocultured with 50 to 250 ?mol/L rutin for 24 h.The inhibition rate of rutin on growth and proliferation of HepG2 was determined by MTT,~(3)H-TdR,and apoptotic cells were observed in fluorescent staining by Olympus fluorescent microscopy,and cell cycle was analysed by flow cytometry.Results Rutin inhibited HepG2 cells from growth and proliferation,and evoked apoptosis.Flow cytometry showed that 50 to 250 ?mol/L rutin caused an increase at G_(0)/G_(1) phase and a decrease at G_(2)/M phase and arrest at G_(0)/G_(1) phase in the cell cycle.Conclusion Rutin markedly inhibits the proliferation of HepG2 cells and induces apoptosis in a concentration-dependent manner.
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Objective To evaluate the intestinal barrier function of the patients by two-sugar absorption test with high performance liquid chromatography (HPLC). Methods Nineteen children with confirmed food hypersensitivity (FH) and 19 normal children were included in this study. The average age was (8.1?1.7) months. After 8-hour fasting, subjects drank test solution (5 g lactulose and 2 g mannitol per 100 ml) at the dose of 2 ml/kg. Five-hour urinary excretion ratio of lactulose/mannitol (L/M) were measured using high performance liquid chromatography (HPLC). The condition of HPLC was as follows: Sugar-PakI column with refractive index detector; Mobile phase: pure deionized water; Flow rate: 0.5 ml/min; temperature of column: 85 ℃. Results HPLC chromatogram of urine sample was stable. The retention time of mannitol and lactulose was 13.86 min and 9.27 min respectively. A significant rise in 5 h urinary L/M excretion ratios was found in children with FH (0.18?0.06) as compared to that of controls (0.05?0.03) (P
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<p><b>BACKGROUND</b>To study the effect of DTB against HIV-1, for developing anti-HIV drugs.</p><p><b>METHODS</b>Different concentration of DTB was added to cell culture system after viral inoculation, MTT staining method for viable cells (MTT assay) and p24 (ELISA) were used as markers to monitor the viral replication.</p><p><b>RESULTS</b>The inhibition rates of DTB at concentrations 160, 80, and 40mg/ml were 93.0%, 56.2% and 18.1%, respectively.</p><p><b>CONCLUSIONS</b>DTB could effectively inhibit HIV-1 in vitro.</p>
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Humans , Anti-HIV Agents , Pharmacology , Bilirubin , Pharmacology , Dose-Response Relationship, Drug , HIV-1 , In Vitro TechniquesABSTRACT
Objective To label the amino-glucomannan(AGM) with fluram and to explore the properties of the compound of fluram-AGM in transmembrane transport.Methods After AGM was labeled with fluram,the peripheral blood mononuclear cells(PBMC) and HepG2 were respectively cultured with Fluram-AGM,then observed under fluorescence microscope with violet light as exciting light.Results Both PBMC and HepG2 showed intracellular indigotic fluorescence.Conclusion AGM can be transported into cells across cell membrane.
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OBJECTIVE: To explore the effectiveness of accident proneness test in drivers. METHODS: Accident proneness test results in 100000 drivers were simulated by function of RANNOR using SAS statistical software, their accident records were simulated by function of UNIFORM according to 3 alternatives, the accident rate of qualified drivers was 0.01, and the accident rate of unqualified drivers was 0.01, 0.05 and 0.10, respectively. RESULTS: It was found that there was no effectiveness of accident proneness test if the accident rate of unqualified drivers was equal to that of the qualified ones, if the accident rate of the unqualified drivers was really higher than that of the qualified ones, the test could identify a certain proportion of high risk drivers. CONCLUSIONS: Accident proneness test may be effective to some extent, it is advisable to carry out further prospective study or pilot work so as to practically prove the effectiveness of the test.
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Objective To investigate the status of infection of human herpes virus type-8 (HHV-8) in the mother and their infants of Wulumuqi and Aletai region. Methods BCBL-1 cell line was used as antigen and 406 matched sera samples were collected from mothers and their infants (cord blood) of different nationalities (Uighur, Kazak and Han nationality) from Wulumuqi and Aletai regions of Xinjiang A.R, tested for HHV-8 IgG and IgM antibody by immunofluorescence assay. Results HHV-8 IgG antibody positive rate of mothers and their infants of Uighur nationality was 22.9%(24/105), Han was 5.4%(8/149) ? 2=17.1,P
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Adult T-cell leukemia/lymphoma (ATLL) and cutaneous T-cell lymphoma (CTCL) have similar clinieopathology and immunophenotype, in order to differentiate the two diseases, 4 eases of ATLL associated cutaneous lesions and 18 eases of CL were studied for elinieopathology, immunophenotype and HTLV-I provirus DNA. Two eases of actinic reticuloid and two eases of lymphocytic infiltration of the skin were selected as negative control. The results showed that four patients of ATLL manifested cutaneous lesions, at same time, they had additional systemic diseases, such as generalized lymphadenopathy, increased levels of LDH and IL-2R, rosette-like cells in their peripheral blood and abnormal bone marrow. The HTLV-I provirus DNA was detected in the peripheral blood, bone marrow, cutaneous lesions and lymph node biopsy specimens of the four patients by PCR amplification of specific HTLV-I DNA fragment. 18 cases of CL were negative for HTLV-I. The study indicates that ATLL may be diagnosed if the patients are associated with CTCL-Iike cutaneous lesions, characteristic histopathological pattern and immunophenotype, rosette-like cells in the peripheral blood and positive HTLV-I provirus DNA.