Epidemiological distribution of genotypes and sub-genotypes of hepatitis B virus in 15 ethnic groups in China / 中华流行病学杂志

Objective: To understand the distribution of genotypes and sub-genotypes of HBV in different ethnic groups in China. Methods: The HBsAg positive samples were selected by stratified multi-stage cluster sampling from the sample base of national HBV sero-epidemiological survey in 2020 for the amplification of S gene of HBV by nested PCR. A phylogeny tree was constructed to determine the genotypes and sub-genotypes of HBV. The distribution of genotypes and sub-genotypes of HBV were analyzed comprehensively by using laboratory data and demographic data. Results: A total of 1 539 positive samples from 15 ethnic groups were successfully amplified and analyzed, and 5 genotypes (B, C, D, I and C/D) were detected. The proportion of genotype B was higher in ethnic group of Han (74.52%, 623/836), Zhuang (49.28%, 34/69), Yi (53.19%, 25/47), Miao (94.12%, 32/34), Buyi (81.48%, 22/27). The proportions of genotype C were higher in ethnic groups of Yao (70.91%, 39/55). Genotype D was the predominant genotype in Uygur (83.78%, 31/37). Genotype C/D were detected in Tibetan (92.35%,326/353). In this study, 11 cases of genotype I were detected, 8 of which were distributed in Zhuang nationality. Except for Tibetan, sub-genotype B2 accounted for more than 80.00% in genotype B in all ethnic groups. The proportions of sub-genotype C2 were higher in 8 ethnic groups, i.e. Han, Tibetan, Yi, Uygur, Mongolian, Manchu, Hui and Miao. The proportions of sub-genotype C5 were higher in ethnic groups of Zhuang (55.56%, 15/27) and Yao (84.62%, 33/39). For genotype D, sub-genotype D3 was detected in Yi ethnic group and sub-genotype D1 was detected in both Uygur and Kazak. The proportions of sub-genotype C/D1 and C/D2 in Tibetan were 43.06% (152/353) and 49.29% (174/353). For all the 11 cases of genotype I infection, only sub-genotype I1 was detected. Conclusions: Five genotypes and 15 sub-genotypes of HBV were found in 15 ethnic groups. There were significant differences in the distribution of genotypes and sub-genotypes of HBV among different ethnic groups.

Lagged effect of temperature on non-accidental mortality and years of life lost in Wuxi: a time-series study / 中华疾病控制杂志

Objective To evaluate the effect of air temperature on non-accidental mortality (A00-R99) and years of life lost in Wuxi city. Methods Data on daily non-accidental mortality and meteorology index were collected from 2012 to 2017. Distributed lag non-linear model (DLNM) was used to assess the effect of temperature on non-accidental death and YLL and the cumulative effects between cold and hot temperature on non-accidental mortality and years of life lost with different lag days. Results A V-shaped relationship was noticed between temperature and mortality. Cold effects were delayed by 3 days and persisted for 14 days. Hot effects appeared acute and reached the peak at the same day. Low temperature had stronger gross effect than high temperature had. There were differences of temperature effects between different age and gender groups. Conclusions Low and high temperature were associated with elevated mortality risk. Cold effect had lagged effect and persisted for long time, however, hot effects appeared acute and the impact of low temperature was greater.

~1H-NMR-based metabonomics analysis of the acute hepatotoxicity mechanism of Artemisia argyi essential oil / 中国中药杂志

This study based on~1H-NMR urine metabolomics technique combined with biochemical indicators to focus on studying the acute hepatotoxicity mechanism of Artemisia argyi essential oil( AAEO). In order to further explore the acute hepatotoxicity mechanism of AAEO,the researchers collected the urine nuclear magnetic data of rats in different periods of high and low doses of olive oil and AAEO group. Using the principal component analysis( PCA) and orthogonal partial least squares-discrimination analysis( OPLSDA) to analyze the endogenous small molecule metabolites in rat urine to study the effects of AAEO on the metabolic process of normal rats. The results showed there was a significant difference between the olive oil group and the AAEO group,the PCA scores chart demonstrated that there was no obvious separation tendency in the urine of olive oil group rats 0-6,6-12,12-24 h,and the metabolic components were distributed in aggregation pattern. The urinary metabolic trajectory of the rats in the AAEO group was conspicuously separated at 0-6,6-12,12-24 h. The experiments proved that the analysis of metabolites by~1H-NMR found that AAEO caused metabolic disorders in rats and produced acute hepatotoxicity. After metabolite differential comparison,it was speculated that the mechanism of acute hepatotoxicity may be involved in the tricarboxylic acid cycle and energy metabolism,while the citrate and oleanolic acid would be the potential biomarkers. This study discussed that the acute hepatotoxicity mechanism of AAEO was used to provide the experimental data for the clinical prescription of Artemisia argyi.

Intranasal Immunization Using CTA1-DD as a Mucosal Adjuvant for an Inactivated Influenza Vaccine / 生物医学与环境科学（英文）

OBJECTIVE@#To evaluate the effect of intranasal immunization with CTA1-DD as mucosal adjuvant combined with H3N2 split vaccine.@*METHODS@#Mice were immunized intranasally with PBS (negative control), or H3N2 split vaccine (3 μg/mouse) alone, or CTA1-DD (5 μg/mouse) alone, or H3N2 split vaccine (3 μg/mouse) plus CTA1-DD (5 μg/mouse). Positive control mice were immunized intramuscularly with H3N2 split vaccine (3 μg/mouse) and alum adjuvant. All the mice were immunized twice, two weeks apart. Then sera and mucosal lavages were collected. The specific HI titers, IgM, IgG, IgA, and IgG subtypes were examined by ELISA. IFN-γ and IL-4 were test by ELISpot. In addition, two weeks after the last immunization, surivival after H3N2 virus lethal challenge was measured.@*RESULTS@#H3N2 split vaccine formulated with CTA1-DD could elicit higher IgM, IgG and hemagglutination inhibition titers in sera. Furthermore, using CTA1-DD as adjuvant significantly improved mucosal secretory IgA titers in bronchoalveolar lavages and vaginal lavages. Meanwhile this mucosal adjuvant could enhance Th-1-type responses and induce protective hemagglutination inhibition titers. Notably, the addition of CTA1-DD to split vaccine provided 100% protection against lethal infection by the H3N2 virus.@*CONCLUSION@#CTA1-DD could promote mucosal, humoral and cell-mediated immune responses, which supports the further development of CTA1-DD as a mucosal adjuvant for mucosal vaccines.

Investigation and analysis of thyroid function of medical occupational population in a tertiary-A hospital in Nanjing / 中华劳动卫生职业病杂志

Objective@#To evaluate the effect of low-dose of ionizing radiation on thyroid function of medical occupational group with long-term exposure, furthermore, to analyze the relationship between the thyroid hormones and the risk factors, such as exposure length, department and comulative radiation dose. Ultimately, providing the scientific basis for setting the ionizing radiation protection standards.@*Methods@#The population who engaged radiodiagnosis and radiotherapy in a tertiary-A hospital was set up as occupational exposure, and the administrative staffs in a company were considered as control. According to the inclusion and exclusion criteria, 161 medical professionals and 159 administrative staffs as the research object.We figured out the basic information and general condition of the groups by face-to-face questionnaire survey, calculated the annual comulative radiation dose through local center for disease control and prevention, By means of the thyroid hormone testing, we analyzed the thyroid hormone levels with different population, occupational exposure factors. Applying Epidata、Excel in data management. All the data was analyzed by statistical software package Stata12.0. Descriptive statistics, single factor analysis of variance and other statistical methods were used for data analysis. Test standard: α=0.05、P<0.05 statistical significant.@*Results@#Age, sex and seniority were proportionality between exposure and control groups. The dosages of occupational population exposure to ionizing radiation were about 1/10 of national permit value, belonging to low-dose exposure. The T3, FT3 level of the exposure group was decreased than the control group (P<0.001). especially the FT3 level has statitical discrepancy among groups with different exposure length (P<0.05).@*Conclusion@#Long-term exposure to low-dose ionizing radiatiom can induce the thyroid damage of medical occupational population, which should be broader concerned.

Cryo-EM structure of an early precursor of large ribosomal subunit reveals a half-assembled intermediate

Assembly of eukaryotic ribosome is a complicated and dynamic process that involves a series of intermediates. It is unknown how the highly intertwined structure of 60S large ribosomal subunits is established. Here, we report the structure of an early nucleolar pre-60S ribosome determined by cryo-electron microscopy at 3.7 Å resolution, revealing a half-assembled subunit. Domains I, II and VI of 25S/5.8S rRNA pack tightly into a native-like substructure, but domains III, IV and V are not assembled. The structure contains 12 assembly factors and 19 ribosomal proteins, many of which are required for early processing of large subunit rRNA. The Brx1-Ebp2 complex would interfere with the assembly of domains IV and V. Rpf1, Mak16, Nsa1 and Rrp1 form a cluster that consolidates the joining of domains I and II. Our structure reveals a key intermediate on the path to establishing the global architecture of 60S subunits.

Effect of low-dose ionizing radiation exposure on thyroid function in a medical occupational population / 中华劳动卫生职业病杂志

Objective@#We evaluated the effect of low-dose of ionizing radiation on thyroid function of medical occupational group with long-term exposure; furthermore; we analyzed the relationship between the thyroid hormones and the risk factors; such as exposure length; department. Ultimately; providing the scientific basis for setting the ionizing radiation protection standards.@*Methods@#The population who engaged radiodiagnosis and radiotherapy in a tertiary-A hospital were set up as occupational exposure; 724 medical professionals as the research object. We figured out the basic information and general condition of the groups by face-to-face questionnaire survey; By means of the thyroid hormone testing; we analyzed the thyroid hormone levels with different population; occupational exposure factors. Then; obtained the prevalence of thyroid nodules by the thyroid ultrasound. Besides; we used the logistic regression model to analyze the risk factors related to thyroid nodule. Applying Epidata、Excel in data management. All the data was analyzed by statistical software package Stata12.0. Descriptive statistics; single factor analysis of variance and other statistical methods were used for data analysis. Test standard: α=0.05、P<0.05 statistical significant.@*Results@#1. Based on the work experience; we divided the study population into four groups; such as 1-9; 10-19; 20-29; and>30 years. The difference of the TSH level among the four groups was statistically significant (P<0.05) . 2. The multiple logistic regression showed that sex and seniority were the independent risk factors for the abnormal rate of thyroid nodules.@*Conclusion@#Long-term exposure to low-dose ionizing radiatiom could induce the thyroid damage of medical occupational population; which should be broader concerned.

Efficient Humoral and Cellular Immune Responses Induced by a Chimeric Virus-like Particle Displaying the Epitope of EV71 without Adjuvant / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To eliminate the side effects of aluminum adjuvant and His-tag, we constructed chimeric VLPs displaying the epitope of EV71 (SP70) without His-tagged. Then evaluating whether the VLPs could efficiently evoke not only humoral but also cellular immune responses against EV71 without adjuvant.</p><p><b>METHODS</b>The fusion protein was constructed by inserting SP70 into the MIR of truncated HBcAg sequence, expressed in E. Coli, and purified through ion exchange chromatography and density gradient centrifugation. Mice were immunized with the VLPs and sera were collected afterwards. The specific antibody titers, IgG subtypes and neutralizing efficacy were detected by ELISA, neutralization assay, and EV71 lethal challenge. IFN-γ and IL-4 secreted by splenocytes were tested by ELISPOT assay.</p><p><b>RESULTS</b>HBc-SP70 proteins can self-assemble into empty VLPs. After immunization with HBc-SP70 VLPs, the detectable anti-EV71 antibodies were effective in neutralizing EV71 and protected newborn mice from EV71 lethal challenge. There was no significant difference for the immune efficacy whether the aluminum adjuvant was added or not. The specific IgG subtypes were mainly IgG1 and IgG2b and splenocytes from the mice immunized produced high levels of IFN-γ and IL-4.</p><p><b>CONCLUSION</b>The fusion proteins without His-tagged was expressed and purified as soluble chimeric HBc-SP70 VLPs without renaturation. In the absence of adjuvant, they were efficient to elicit high levels of Th1/Th2 mixed immune response as well as assisted by aluminum adjuvant. Furthermore, the chimeric VLPs have potential to prevent HBV and EV71 infection simultaneously.</p>

Look into Hepatic Progenitor Cell Associated Trait: Histological Heterogeneity of Hepatitis B-Related Combined Hepatocellular-Cholangiocarcinoma / 华中科技大学学报（医学）（英德文版）

Combined hepatocellular-cholangiocarcinoma (CHC) is a mixed tumor containing elements of both hepatocellular carcinoma (HCC) and cholangiocarcinoma (CC).Its remarkable histological heterogeneity has been linked to putative hepatic progenitor cell (HPC) origin.However,detailed histological or phenotypic description is rarely documented.In the present study,we reassessed 68 cases previously diagnosed as hepatitis B-related CHCs by immunohistochemistry and double-fluorescence immunostaining,focusing on HPC associated phenotypic observation of intermediate area of the tumor.It was found that tumor cells showed remarkable heterogeneity in intermediate area.Tumor cells with intermediate morphology between hepatocytes and cholangiocytes were oval-shaped and small with scant cytoplasm and hyperchromatic nuclei,arranging in solid nests mostly.By Keratin 7 (K7) staining,it appeared that the nests of tumor cells represented a maturation process from the undifferentiated small cells to mature hepatocytes through the "transitional" cells.Then,these small cells were further confirmed with intermediate phenotype as HPC by exploring immature hepatocellular marker and HPC/biliary markers co-localization.In conclusion,the HPC associated trait in CHC can be interpreted by HPC origin or gain of"stemness" by dedifferentiation.It is still too soon to give a final word that it is innate or acquired signature of HPC associated trait in CHC.

Association between Plasma Metal Levels and Diabetes Risk: a Case-control Study in China / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>Many metals, some of which have been classified as environmental endocrine disruptors, are used extensively in everyday consumer products and are ubiquitous in our living environment. In the present study, we aimed to explore the associations between the prevalence risk of type 2 diabetes and plasma levels of 20 trace elements as well as those of heavy metals in a Han Chinese population.</p><p><b>METHODS</b>We conducted a case-control study to investigate the associations between plasma concentrations of 20 metals and diabetes in Jiangsu province. A total of 122 newly diagnosed cases of type 2 diabetes and 429 matched controls were recruited from community physical examinations in Suzhou City of Jiangsu Province. Plasma metal levels were measured by inductively-coupled plasma mass spectrometry.</p><p><b>RESULTS</b>After adjusting for confounders, plasma vanadium, chromium, manganese, copper, zinc, arsenic, selenium, strontium, palladium, cadmium, cesium, and barium were associated with diabetes risk (P < 0.05). The adjusted OR increased with increasing concentration of vanadium, manganese, copper, zinc, and cesium.</p><p><b>CONCLUSION</b>Many metals, including manganese, copper, zinc, arsenic, selenium, and cadmium in plasma, are associated with the morbidity of diabetes. Monitoring of environmental metal levels and further studies are urgently needed.</p>

Meta-analysis for efficacy and safety of rosuvastatin and simvastatin in the treatment of hyperlipidemia / 天津医药

Objective To investigate the effectiveness and safety of rosuvastatin and simvastatin in the treatment of hyperlipidemia. Methods The database including CNKI, VIP, Wanfang data base and CBM were retrieved to search the clinical randomized controlled trials (RCT) of rosuvastatin and simvastatin in the treatment of hyperlipidemia, and the data were analyzed with Review Manager 5.2. Results Eighteen RCTs were included with a total sample size of 1819 cases with hyperlipidemia, in which there were 917 patients in rosuvastatin group and 902 in simvastatin group. The Meta-analysis results showed that there were significantly lower serum levels of total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), triglyceride (TG) and significantly higher level of high-density lipoprotein cholesterol (HDL-C) in rosuvastatin group compared with those of simvastatin group [(MD=-0.15, 95%CI:-0.22--0.09, P<0.01), (MD=-0.18, 95%CI:-0.25--0.11, P<0.01), (MD=-0.23, 95%CI:-0.28--0.19, P<0.01) and (MD=-0.11, 95%CI:-0.06--0.15, P<0.01)]. There was no significant difference in the incidence of gastrointestinal adverse reaction between the two groups. Conclusion The current clinical evidences show that rosuvastatin has a better effect on the treatment of hyperlipidemia, and has no adverse reactions.

Expression and Purification of Recombinant Hepatitis Delta Virus (HDV) Antigen for Use in a Diagnostic ELISA for HDV Infection Using the High-Density Fermentation Strategy in Escherichia coli / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>Hepatitis Delt a Virus (HDV) antigen is widely used as a capture antigen in ELISAs for the identification of HDV infection; large amounts of recombinant HDV antigen with active antigenicity are required for this purpose.</p><p><b>METHODS</b>Reconstruct the gene of HDV antigen based on the bias code of Escherichia coli, the recombinant protein expresses by high-density fermentation with fed-batch feeding strategy, and purify by immobilized metal chromatography. The sensitivity and specificity of this antigen detect by ELISA method.</p><p><b>RESULTS</b>The expression of HDV antigen can reach 20% of the total cell mass in the soluble form. The recombinant HDV antigen can be conveniently purified (98%) by immobilized metal ion affinity chromatography (IMAC) using the interaction between a His-tag and nickel ions. Production of recombinant HDV antigen can reach 0.5 g/L under conditions of high-density cell fermentation. Applied to the diagnostic ELISA method, the recombinant HDV antigen shows excellent sensitivity (97% for IgM and 100% for IgG) and specificity (100% for IgG and IgM) for the detection of anti-HDV antibodies.</p><p><b>CONCLUSION</b>Expression and purification the recombinant HDV antigen as a candidate protein for application in a diagnostic ELISA for HDV infection. Large-scale production of the protein can be achieved using the high-density fermentation strategy.</p>

Association between chemical composition of essential oil with penetration enhancement effect and drug properties of traditional Chinese medicine / 中国中药杂志

The results of previous studies showed potential correlations between the penetration enhancement effect of essential oils and the drug properties of traditional Chinese medicine based on the data mining method. As chemical composition is the material basis of drug properties of traditional Chinese medicine, this article further analyzed the correlation between the chemical composition of essential oils and the drug properties. Firstly, essential oils were extracted by steam distillation, and then physicochemical parameters of essential oils, such as relative density and refractive index, were measured. The chemical components of 20 essential oils were analyzed by GC-MS, and divided into 12 categories according to skeleton features and functional groups. Finally, Logistic regression analysis was applied to reveal the correlations. The results proved that five flavors, four tastes and channel tropisms showed the correlation with chemical composition of essential oils (P<0.05). In conclusion, there were obvious correlations and regularity between the drug properties of traditional Chinese medicine and the chemical composition of essential oils.

Evaluation of pharmacokinetics and in vitro/in vivo correlation of ibuprofen with essential oils as penetration enhancer following transdermal administration / 中国中药杂志

The study was carried out to investigate the pharmacokinetics and in vitro/in vivo correlation of ibuprofen with essential oils as penetration enhancers (PE) following transdermal administration. With Azone as the positive control, ibuprofen hydrogels containing Chuanxiong oil, Angelica oil or Cinnamon oil as PE were prepared and administered to the rat abdominal skin. Then the pharmacokinetics of ibuprofen following transdermal administration were investigated and compared. In comparison with negative control (no PE was added), the relative bioavailability values with the addition of Chuanxiong oil, Angelica oil, Cinnamon oil and Azone as PE were determined to be 161.87%, 171.05%, 151.37% and 148.66%, respectively. In vitro/in vivo correlation analysis was performed by deconvolution method and the results demonstrated a good correlation between in vitro and in vivo percutaneous absorption studies. The correlation coefficients were measured to be 0.999 7, 0.995 2 and 0.999 4 for Chuanxiong oil, Angelica oil and Cinnamon oil respectively. In summary, Chuanxiong oil, Angelica oil and Cinnamon oil as PE could significantly enhance the bioavailability of ibuprofen following transdermal administration. A satisfactory in vitro/in vivo correlation could be obtained by using hydrogel as the dosage form.

Structural characterization of coatomer in its cytosolic state

Studies on coat protein I (COPI) have contributed to a basic understanding of how coat proteins generate vesicles to initiate intracellular transport. The core component of the COPI complex is coatomer, which is a multimeric complex that needs to be recruited from the cytosol to membrane in order to function in membrane bending and cargo sorting. Previous structural studies on the clathrin adaptors have found that membrane recruitment induces a large conformational change in promoting their role in cargo sorting. Here, pursuing negative-stain electron microscopy coupled with single-particle analyses, and also performing CXMS (chemical cross-linking coupled with mass spectrometry) for validation, we have reconstructed the structure of coatomer in its soluble form. When compared to the previously elucidated structure of coatomer in its membrane-bound form we do not observe a large conformational change. Thus, the result uncovers a key difference between how COPI versus clathrin coats are regulated by membrane recruitment.

4.4 Å Resolution Cryo-EM structure of human mTOR Complex 1

Mechanistic target of rapamycin (mTOR) complex 1 (mTORC1) integrates signals from growth factors, cellular energy levels, stress and amino acids to control cell growth and proliferation through regulating translation, autophagy and metabolism. Here we determined the cryo-electron microscopy structure of human mTORC1 at 4.4 Å resolution. The mTORC1 comprises a dimer of heterotrimer (mTOR-Raptor-mLST8) mediated by the mTOR protein. The complex adopts a hollow rhomboid shape with 2-fold symmetry. Notably, mTORC1 shows intrinsic conformational dynamics. Within the complex, the conserved N-terminal caspase-like domain of Raptor faces toward the catalytic cavity of the kinase domain of mTOR. Raptor shows no caspase activity and therefore may bind to TOS motif for substrate recognition. Structural analysis indicates that FKBP12-Rapamycin may generate steric hindrance for substrate entry to the catalytic cavity of mTORC1. The structure provides a basis to understand the assembly of mTORC1 and a framework to characterize the regulatory mechanism of mTORC1 pathway.

Structural dynamics of the yeast Shwachman-Diamond syndrome protein (Sdo1) on the ribosome and its implication in the 60S subunit maturation

The human Shwachman-Diamond syndrome (SDS) is an autosomal recessive disease caused by mutations in a highly conserved ribosome assembly factor SBDS. The functional role of SBDS is to cooperate with another assembly factor, elongation factor 1-like (Efl1), to promote the release of eukaryotic initiation factor 6 (eIF6) from the late-stage cytoplasmic 60S precursors. In the present work, we characterized, both biochemically and structurally, the interaction between the 60S subunit and SBDS protein (Sdo1p) from yeast. Our data show that Sdo1p interacts tightly with the mature 60S subunit in vitro through its domain I and II, and is capable of bridging two 60S subunits to form a stable 2:2 dimer. Structural analysis indicates that Sdo1p bind to the ribosomal P-site, in the proximity of uL16 and uL5, and with direct contact to H69 and H38. The dynamic nature of Sdo1p on the 60S subunit, together with its strategic binding position, suggests a surveillance role of Sdo1p in monitoring the conformational maturation of the ribosomal P-site. Altogether, our data support a conformational signal-relay cascade during late-stage 60S maturation, involving uL16, Sdo1p, and Efl1p, which interrogates the functional P-site to control the departure of the anti-association factor eIF6.

Association between penetration enhancement effect of essential oils and drug properties of traditional Chinese medicines by data mining method / 中国中药杂志

To study the association between penetration enhancement effect of essential oils and drug properties of traditional Chinese medicines. Through literature research, 34 kinds of essential oils with the penetration enhancement effect were collected. The methods of frequency analysis and variable crosstab were used for intuitive analysis and association analysis. The association between penetration enhancement effect of essential oils and drug properties (four natures, five flavors, channel tropism) were analyzed by a general linear model. According to the findings, the essential oils with penetration enhancement effect were all sourced from acrid traditional Chinese medicines, because their positive drug nature contributed to the enhancement of the penetration effect of essential oil; five flavors had little effect on penetration enhancement (P = 0.6982), but four natures and channel tropism showed significant effects (P = 0.011, 0.077). In conclusion, there were obvious association and regularity between penetration enhancement effect of essential oils and drug properties of traditional Chinese medicine.

Prokaryotic soluble expression of protein D of Haemophilus influenzae type b / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To express the recombinant D protein in prokaryotic expression system solubly and make preparation for producing D-carrier conjugate vaccine next step.</p><p><b>METHODS</b>The hpd gene fragment removed of signal peptide from genomic DNA of Hib CMCC was inserted into pET43. 1a. The recombinant plasmid was transformed to competent E. coli BL21 (DE3) for expression under induction of IPTG. The expressed recombination protein was precipitated with ammonium sulfate, purified by DEAE anion exchange column chromatography and identified for reactogenicity by Western Blot.</p><p><b>RESULTS</b>The expressed recombination protein, in a soluble form, constained about 50% of total somatic protein and showed specific reaction with the HIB antisera after preliminary purification.</p><p><b>CONCLUSION</b>The D protein recombined expression plasmid was constructed successfully and expressed D protein in prokaryotic cells in a solube form.</p>

Construction of recombinant CHO cell strain for high expression of HBsAg / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To overexpress hepatitis B virus S gene in CHO cells cultured in serum-free media.</p><p><b>METHOD</b>Plasmid was constructed by cloning of HBV S gene and then it was transfected into CHO cells. After cell screen, the positive clones were identified and isolated into a serum-free media followed by the serological and morphological characterization of the expression product.</p><p><b>RESULT</b>CHO cell strains which can express HBsAg efficiently and stably were obtained. Spherical and filamentous HBsAg could be detected under electronic microscope. The titer of the expression product was up to 1:5000.</p><p><b>CONCLUSION</b>Serum-free media cultured CHO cell strain for overexpression of HBsAg was successfully constructed and the expression product was high antigenic.</p>