Correlation of Moraxella catarrhalis macrolide susceptibility with the ability to adhere and invade human respiratory epithelial cells.

Recently, the prevalence of macrolide-resistant Moraxella catarrhalis has been reported, especially among Chinese children. The fitness cost of resistance is reported to render the resistant bacteria less virulent. To investigate the correlation between macrolide susceptibility of M. catarrhalis and pathogenicity, the whole genome of 70 M. catarrhalis isolates belonging to four clonal complexes with different macrolide susceptibilities was sequenced. The gene products were annotated with the Gene Ontology terms. Based on 46 extracted essential virulence genes, 19 representative isolates were selected to infect type II alveolar cells (A549 cells). The ability of these isolates to adhere and invade human epithelial cells and to produce cytokines was comparatively analysed. Furthermore, mice were infected with a pair of M. catarrhalis isolates with different pathogenic behaviours and macrolide susceptibilities to examine pulmonary clearance, histological findings, and the production of cytokines. The percentages of annotations for binding, metabolic process, cellular process, and cell were non-significantly different between the macrolide-resistant and macrolide-susceptible groups. The presence of uspA2, uspA2H, pilO, lbpB, lex1, modM, mboIA, and mboIB significantly differed among the four clonal complexes and macrolide susceptibility groups. Furthermore, compared with those in macrolide-susceptible isolates, the adhesion ability was stronger (P = 0.0019) and the invasion ability was weaker (P < 0.0001) in the macrolide-resistant isolates. Mouse experiments revealed that pulmonary macrophages elicit immune responses against M. catarrhalis infection by significantly upregulating the Csf2, Il4, Il13, Il1b, Il6, Tnf, and Il18. Therefore, M. catarrhalis populations exhibited diverse pathogenicity in vitro and in vivo.

Histone deacetylase inhibitor induced pVHL-independent degradation of HIF-1α and hierarchical quality control of pVHL via chaperone system.

The mortality rate of ovarian cancer is increasing and the role of hypoxia inducible factor-1α (HIF-1α) in tumor progression has been confirmed. von Hippel-Lindau tumor suppressor protein (pVHL) binds HIF-1α and mediates proteasome degradation of HIF-1α. Besides, histone deacetylase inhibitor (HDACi) mitigates tumor growth via targeting HIF-1α, whereas underlying mechanism still requires investigation. In this research, we exposed ovarian cancer cell lines OV-90 and SKOV-3 to escalating concentrations of HDACi LBH589. As a result, cell viability was significantly suppressed and expression of HIF-1α was remarkably reduced along with decreased levels of signal molecules, including phosphoinositide 3-kinase (PI3K) and glycogen synthase kinase 3ß (GSK3ß) (P = 0.000). Interestingly, pVHL was expressed in a notably declining tendency (P = 0.000). Chaperone heat shock protein-70 (HSP70) was expressed in an ascending manner, whereas expression of chaperonin TCP-1α was reduced clearly (P = 0.000). Besides, co-inhibition of pVHL plus HDAC did not contribute to a remarkable difference in HIF-1α expression as compared with single HDAC inhibition. Furthermore, both cell lines were transfected with plasmids of VHL plus VHL binding protein-1 (VBP-1). Consequently, the expression of HIF-1α as well as lactate dehydrogenase-A (LDHA) was remarkably decreased (P = 0.000). These findings indicate HDACi may repress expression of HIF-1α via inhibiting PI3K and GSK3ß and promote degradation of HIF-1α via HSP70, independent of pVHL. Additionally, a sophisticated network of HDAC and chaperones may involve in pVHL quality control.

Hepatitis B virus serological screen in a general hospital in Beijing from 2008 to 2018, and challenges to our vaccination policy.

OBJECTIVE: Hepatitis B virus (HBV) has a worldwide distribution and remains a leading public health problem in China. METHOD: Automated chemiluminescence microparticle immunoassay was used to test all five markers of HBV serology in serum samples among 696,048 patients, pregnant women, and normal subjects in Beijing from 2008 to 2018. RESULTS: The overall prevalence of subjects categorized as previous/ occult HBV infection, inactive HBsAg carrier, active HBV infection, HBsAg, HBV susceptible, and immune via vaccination was 29.4%, 4.8%, 1.4%, 6.4%, 33.9% and 30.3%, respectively; men had a significantly higher prevalence of HBV infection than women. The prevalence of HBsAg was around 0.5% in subjects ≤ 10 years of age, increased dramatically to 3.7% in subjects between 11 and 20 years of age, reached the highest level of 7.9% in subjects between 41 and 50 years of age, and finally decreased to 2.8% in subjects ≥ 81 years of age. During the 10 years from 2008 to 2018, the prevalence of HBsAg was stabilized at about 6.0%, and indicators of HBV susceptibility, previous/ occult HBV infection, and immunity via vaccination were not further improved, despite the constant implementation of HBV vaccination since 1992. All four age groups (21 - 30y, 31 - 40y, 41 - 50y and 51 - 60y) of the normal adult population were found to have a significantly lower prevalence of HBsAg and HBV susceptibility but significantly higher prevalence of immunity via vaccination compared with corresponding age groups of the sub-total population. CONCLUSIONS: Although high coverage has been established among infants and young children, their vaccination alone could not reduce HBV infection in the adult Chinese population quickly. Adult populations with more vaccinated individuals are found to have fewer individuals with HBsAg. Vaccination in adults or at least in high-risk adults is an urgent need to decrease horizontal HBV transmission in China.

Cytomegalovirus antigenemia in patients with autoimmune and non-autoimmune diseases in Beijing: A 10-year single hospital experience.

BACKGROUND: Primary cytomegalovirus (CMV) infection is prevalent worldwide and usually results in latency in immunocompetent populations. Reactivation of latent CMV can cause life-threatening complications in immunocompromised hosts. METHODS: We used the CMV Brite assay to test CMV antigenemia (pp65) in whole blood samples from 22,192 patients with or without autoimmune diseases in Beijing during 2008-2018. RESULTS: The overall prevalence of CMV antigenemia was 19.5% (9.7%, males; 26.0%, females). The prevalence of CMV antigenemia was 35.1%, 58.6% and 11.4% in whole patients with autoimmune diseases, in patients with systemic lupus erythematosus (SLE) and in patients with non-SLE autoimmune diseases, respectively. All patients with non-autoimmune diseases, patients with HIV/AIDS or transplantation were found to have 5.0%, 27% or 14.8%, respectively. Patients≤20 years with SLE had a significantly higher prevalence of CMV antigenemia than did all SLE patients, on average. Patients>51 years with non-SLE autoimmune diseases had a significantly higher prevalence than did all patients with non-SLE autoimmune diseases, on average. The prevalence of CMV antigenemia in patients admitted to intensive-care units (ICUs) were 9.2%, which was significantly higher than that among all patients with non-autoimmune diseases. Patients with SLE had 23.8% of negative conversion of CMV antigenemia, significantly lower than the percentage of patients with non-SLE autoimmune (64.3%) and non-autoimmune (61.0%) diseases. The mean number of days to negative conversion of CMV antigenemia in patients with SLE was 35.3±35.8 days, which was significantly longer than that in patients with non-SLE autoimmune diseases (15.4±11.9 days) and non-autoimmune diseases (13.6±7.7 days). CONCLUSIONS: CMV antigenemia is found more likely in women than in men, more prevalently in patients with SLE than those with HIV/AIDS or transplant recipients, more frequently in patients admitted to ICUs. Patients with SLE had prolonged CMV antigenemia. The role of CMV appears important in SLE.

A retrospective seroepidemiologic survey of Chlamydia pneumoniae infection in patients in Beijing between 2008 and 2017.

BACKGROUND: Chlamydia pneumoniae (C. pneumoniae) is an obligate intracellular bacterium and a human pathogen that causes respiratory infectious diseases. More than 50% of the adult population worldwide was once infected with C. pneumoniae, but investigations into this topic are insufficient in mainland China. METHODS: Anti-C. pneumoniae IgG and IgM antibodies were detected using micro-immunofluorescence test in serum samples of patients visiting Peking Union Medical College Hospital between 2008 and 2017 for routine medical purposes, and the aim of this retrospective study was to analyze the test results. RESULTS: Among 12,050 serum specimens tested for anti-C. pneumoniae IgG and IgM antibodies, the overall prevalence of anti-C. pneumoniae IgG antibodies was 86.6%, 87.2% for men and 86.0% for women. Adult men (>20 years) were found to have a significantly higher prevalence of anti-C. pneumoniae IgG than women (χ2 = 30.32, P = 0.000). 3 to 5 years old patients were observed to have the lowest prevalence of anti-C. pneumoniae IgG, 42.8%, then increased with age, reaching the highest level of 98.6% in patients over 70 years of age. In the 10,434 specimens with C. pneumoniae IgG antibodies, the total geometric mean titer (GMT) for C. pneumoniae IgG was 45.71. Although GMTs were found to be significantly higher among all men than among all women (t = 5.916, P = 0.000), sex difference actually began in patients over 40 years of age and increased in the elderly. In the total 12,050 specimens, 1.2% had anti-C. pneumoniae IgM, 3.3% had anti-C. pneumoniae IgG with titers equal to or greater than 1:512; 0.39% had ≥4-fold increasing titers of antibodies in acute and convalescent phase paired samples, and 4.4% were finally confirmed to have acute antibodies against C. pneumoniae. 6 to 10 years old patients were found to have the highest rate of both IgM antibodies (3.9%) and acute antibodies (6.2%) against C. pneumoniae. Acute antibodies against C. pneumoniae were found to be more frequent in patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD, 14.0%, χ2 = 20.43, P = 0.000), patients with pneumonia (7.8%, χ2 = 51.87, P = 0.000) and patients with acute respiratory tract infection (12.3%, χ2 = 60.91, P = 0.000) than among all patients (4.4%). Both anti-C. pneumoniae IgG and IgM antibodies should be tested for acute antibodies against C. pneumoniae as testing for either alone will underestimate by a maximum of two-thirds the incidence of acute antibodies against C. pneumoniae. CONCLUSIONS: More than 86% of Chinese patients on an average were once infected with C. pneumoniae. Adult men had both a higher prevalence and higher levels of antibodies than women. 6 to10 year old patients were found to have the most frequent acute infection of C. pneumoniae. C. pneumoniae is associated with AECOPD, pneumonia and acute respiratory tract infection. Anti-C. pneumoniae IgG and IgM should be tested simultaneously to avoid underestimation of acute antibodies against C. pneumoniae.

Landscape of PD-1/PD-L1 Regulation and Targeted Immunotherapy.

Programmed cell death protein 1 (PD-1)/programmed death ligand 1 (PD-L1) is a significant immune checkpoint, and the dysfunction of this axis contributes to tumor metastasis and immune escape. PI3K/Akt/mTOR and MAPK signal network induces PD-1/PD-L1 expression and facilitates tumor progression. Transcriptional factors such as hypoxia induced factors, PTEN, p53, CDK5, BRD4, STAT modulate PD-1/PD-L1 expression. PD-1/PD-L1 level is also regulated via epigenetic and post-translational manner. The underlying mechanisms mentioned above may provide potential targets for tumor treatment. At present, the combination therapy of PD-1/PD-L1 monoclonal antibodies plus small molecular inhibitors has achieved good outcomes in tumor treatment.

Anti-Epstein-Barr virus antibodies in Beijing during 2013-2017: What we have found in the different patients.

BACKGROUND: Epstein-Barr virus (EBV) is associated with nasopharyngeal carcinoma (NPC) which is prevalent in South China, and its association with systemic lupus erythematosus (SLE) or other autoimmune diseases has not been studied in the mainland of China. The EBV serological tests have been performed on patients with various diseases or manifestations for years at our institution and their values need to be evaluated. METHODS: For routine medical purposes, anti-EB viral capsid antigen (VCA) IgG, IgA and IgM antibodies, anti-EBV diffuse early antigen (EA-D) IgA antibodies, and anti-EBV nuclear antigen-1(EBNA-1) IgG antibodies were tested with commercial enzyme-linked immunosorbent assay (ELISA) in patients visiting Peking Union Medical College Hospital between 2013 and 2017. The test results were analyzed in this retrospective study. RESULTS: There were a total of 11122 serum samples available to be tested in the study. As indicators of past EBV infection, the prevalence of VCA-IgG/EBNA1-IgG were 66.6%/58.5%, 84.3%/78.8%, 92.9%/87.0% and 98.5%/95.4% in patients aged under 5 years, 6-10 years, 11-20 years and 21-30 years old, respectively, and these values maintained at this highest rate as age increased further. The prevalence of VCA-IgM, as a parameter of acute EBV infection, was 14.6%, 10.2%, 10.4%, 6.3% and 3.1% in patients aged under 5 years, 6-10 years,11-20 years, 21-30 years, 31-40 years old, respectively, and decreased to 2%~3% in older patients. Patients with elevated serum liver enzymes were more likely to have a higher prevalence of EA/D IgA antibody (P < 0.01) and young patients (≤30 years) with lymphadenopathy were more likely to have higher prevalence of VCA-IgM antibody (P < 0.01). The prevalence of VCA-IgA and EAD-IgA were 87.0% and 59.2% in NPC patients, respectively, and both were significantly higher (P < 0.001) than that in non-NPC patients. The prevalence of VCA-IgA was 45.4% and 25.6% in SLE patients and patients with other autoimmune diseases, respectively, which were significantly (P < 0.001) and mildly (P = 0.039) higher than their controls. In pediatric SLE patients between 6 and10 years old, the prevalence of VCA-IgG, VCA-IgA and EBNA1-IgG was 100%, 59.5% and 100%, respectively, all being significantly higher than the age (6-10y) related controls (P< 0.01). In the 705 cerebral spinal fluid (CSF) specimens, VCA-IgG, VCA-IgM, VCA-IgA and EAD-IgA were found to be positive in 12.1%, 0.15%, 0.25% and 0.25%, respectively. There were 157 paired specimens (CSF and serum were collected simultaneously) and VCA-IgG was identified as positive in 12.7% of the CSF and 100% of the serum specimens. CONCLUSIONS: Around 98% of Chinese patients were infected with EBV before 30 years of age and the highest rate of acute EBV infection were observed in patients under 5 years old. EBV infection was found to be associated with elevated serum liver enzymes, NPC and SLE. Acute anti-EBV antibody was valued for young patients with lymphadenopathy but limited value for CNS neuropathy.

Alteration of glucocorticoid receptors and exacerbation of inflammation during lytic cytomegalovirus infection in THP-1 cells.

Cytomegalovirus (CMV) infection is associated with glucocorticoid resistance in ulcerative colitis (UC) and may exacerbate the disease course. However, the underlying pathogenicity remains unclear. The aim of this study was to explore possible underlying mechanisms during CMV latency and lytic infection in the human mononuclear cell line THP-1. Latent and activated CMV infection cell models were established. We performed real-time PCR and western blotting to examine changes in glucocorticoid receptors (GRs) during CMV latency and activation. Pro-inflammatory and anti-inflammatory cytokines were detected by ELISA. After UV-inactivated CMV infection, GRs and cytokines were also examined. The expression of GRs was elevated in the reactivation group. An increased ratio of GR ß/α and phosphorylation of GRα in the CMV reactivation group may explain refractory response to steroids. During CMV lytic infection, pro-inflammatory cytokines IL-6 and TNF-α increased remarkably and anti-inflammatory cytokine IL-5 decreased, which may exacerbate UC. GR and cytokines were unchanged in the UV-inactivated CMV infection group. Changes in the number and function of GRs may account for glucocorticoid resistance in CMV reactivation. The imbalance of pro- and anti-inflammatory cytokines may be related to severe inflammation.

Seroprevalence of the Hepatitis B, Hepatitis C, and Human Immunodeficiency Viruses and Treponema pallidum at the Beijing General Hospital from 2010 to 2014: A Cross-Sectional Study.

BACKGROUND: The hepatitis B, hepatitis C, human immunodeficiency viruses and Treponema pallidum are important causes of infectious diseases concern to public health. METHODS: Between 2010 and 2014, we used an automated chemiluminescence microparticle immunoassay to detect the hepatitis B, hepatitis C, and human immunodeficiency viruses as well as Treponema pallidum (the rapid plasma regain test was used in 2010-2011). Positive human immunodeficiency virus tests were confirmed via western blotting. RESULTS: Among 416,130 subjects, the seroprevalences for hepatitis B virus, hepatitis C virus, human immunodeficiency virus, and Treponema pallidum were 5.72%, 1.23%, 0.196%, and 0.76%, respectively. Among 671 patients with positive human immunodeficiency virus results, 392 cases were confirmed via western blotting. Hepatitis B and human immunodeficiency virus infections were more frequent in men (7.78% and 0.26%, respectively) than in women (4.45% and 0.021%, respectively). The hepatitis B and C virus seroprevalences decreased from 6.21% and 1.58%, respectively, in 2010, to 5.37% and 0.988%, respectively, in 2014. The human immunodeficiency virus seroprevalence increased from 0.04% in 2010 to 0.17% in 2014, and was elevated in the Infectious Disease (2.65%), Emergency (1.71%), and Dermatology and Sexually Transmitted Diseases (1.12%) departments. The specificity of the human immunodeficiency virus screening was 71.4%. The false positive rates for the Treponema pallidum screening tests increased in patients who were 60-70 years old. The co-infection rates for the hepatitis C and human immunodeficiency viruses were 0.47% in hepatitis C virus-positive patients and 7.33% in human immunodeficiency virus-positive patients. CONCLUSIONS: During 2010-2014, hepatitis B virus and human immunodeficiency virus infections were more frequent among men at our institution. Although the seroprevalences of hepatitis B and C viruses decreased, the seroprevalence of human immunodeficiency virus infection increased (with higher seroprevalences in high-risk departments). Older patients were more likely to exhibit false positive findings for syphilis.

[Detection of serum Chlamydia pneumoniae antibodies and analysis of epidemiologic data in 5 years].

OBJECTIVE: To detect serum Chlamydia pneumoniae (C.pneumonia) antibodies and analyze the epidemiologic data. METHODS: Micro-immunofluorescence (MIF) was used to detect IgG and IgM antibodies against C.pneumoniae in 7 802 serum specimens from May 2008 to September 2013. C.pneumoniae past or recent infections were determined based on specific IgG and IgM antibodies and their titers. The analysis of relationship between age, gender and other clinical factors associated with antibody titers was performed. RESULTS: The positive rate of C.pneumonia IgG antibodies ( ≥ 1: 16 and <1: 512) was 87.5% (6 827/7 802). And the lowest rate of 50.6% (78/154) was shown in patients of 2-4 years age group while the highest rate of 98.7% (1 277/1 294) in those above 70 years old. The average geometric mean titer (GMT) of C.pneumonia IgG antibodies was 43.9. GMT in males was significantly higher than that in females (45.8 vs 41.9, P = 0.000 1). And 282 (3.6%) patients were diagnosed with recent C.pneumoniae infection (IgG ≥ 1: 512 or IgM ≥ 1: 16 or acute and convalescent serum antibody titers turned to positive or four times and more elevated). The lowest rate of 0.5% (2/417) was detected in < 2-year-old patients while the highest rate of 6.7% (24/357) in 10-14-year-old group patients. The miss ratio was as high as 68.1% (192/282) if C.pneumoniae IgM antibody was used as an only criteria for C. pneumoniae recent infection. The recent infection rates in patients with autoimmune diseases (7.1% (24/336) ) and those with pneumonia/chest radiological shadow findings (4.7% (69/1 467)) were statistically higher than the average rate (3.6%) of total population (P = 0.000 4, 0.014 0). CONCLUSIONS: The positive rate of C.pneumonia IgG antibody is quite high in the population. And the GMT of C.pneumoniae IgG antibody in males is significantly high than that in females. Recent C.pneumonia infection is to be missed if IgM antibody is used as a sole criterion.

[Analysis of cytomegaloviral antigenemia test and re-test after antiviral chemotherapy in patients with autoimmune and non-autoimmune diseases].

OBJECTIVE: To retrospectively analyze the cytomegalovirus (CMV) antigenemia test and re-test after antiviral chemotherapy in patients with autoimmune and non-autoimmune diseases. METHODS: CMV Brite kit and indirect immunofluorescence were used to detect CMVpp65 antigenemia in 6471 peripheral blood leukocyte specimens from 5325 clinic and hospitalized patients with clinically suspicious CMV infections from May 2008 to February 2012. And the positive results were defined as episodes of systemic CMV activity. RESULTS: In 6471 EDTA-treated peripheral blood specimens, 948 (14.6%) were found with positive CMV antigenemia. The average positive rate from 13 kinds of autoimmune diseases was 34.9% (670/1922) in which systemic lupus erythematosus patients had the highest (52.4%, 551/1052). Meanwhile, the average positive rate from 12 kinds of non-autoimmune diseases was only 6.1% (144/2367) in which it was 17.3% (27/156) in patients with respiratory/acute renal failure, acquired immunodeficiency syndrome (AIDS) and kidney transplant recipients. And 189 patients with positive antigenemia were re-tested after antiviral chemotherapy and only 64 (33.9%) were converted negatively. CONCLUSIONS: Patients with autoimmune diseases have replaced traditionally immunocompromised patients, e.g. AIDS and kidney transplant recipient, to become the highest risk group of systemic CMV activity. Negative conversion rate of CMV antigenemia is low after antiviral chemotherapy.

Construction of a multiple myeloma diagnostic model by magnetic bead-based MALDI-TOF mass spectrometry of serum and pattern recognition software.

A diagnosis of multiple myeloma (MM) is difficult to make on the basis of any single laboratory test result. Accurate diagnosis of MM generally results from a number of costly and invasive laboratory tests and medical procedures. The aim of this work is to find a new, highly specific and sensitive method for MM diagnosis. Serum samples were tested in groups representing MM (n = 54) and non-MM (n = 108). These included a subgroup of 17 plasma cell dyscrasias, a subgroup of 17 reactive plasmacytosis, 5 B cell lymphomas, and 7 other tumors with osseus metastasis, as well as 62 healthy donors as controls. Bioinformatic calculations associated with MM were performed. The decision algorithm, with a panel of three biomarkers, correctly identified 24 of 24 (100%) MM samples and 46 of 49 (93.88%) non-MM samples in the training set. During the masked test for the discriminatory model, 26 of 30 MM patients (sensitivity, 86.67%) were precisely recognized, and all 34 normal donors were successfully classified; patients with reactive plasmacytosis were also correctly classified into the non-MM group, and 11 of the other patients were incorrectly classified as MM. The results suggested that proteomic fingerprint technology combining magnetic beads with MALDI-TOF-MS has the potential for identifying individuals with MM. The biomarker classification model was suitable for preliminary assessment of MM and could potentially serve as a useful tool for MM diagnosis and differentiation diagnosis.

[Analysis of the correlation between PreS1-Ag, PreS2-Ag, HBeAg and HBV DNA in the serum of chronic hepatitis B patients].

OBJECTIVE: To investigate the correlation between the serum indices and HBV DNA. METHODS: 100 chronic HBV patients and 40 healthy controls were enrolled in this study. The patients have not received any treatment with interferon (IFN) and similar nucleotide, and featured with normal ALT/AST, positive HBV DNA examined by using internal-quantitative standard PCR. Serum indices were measured by micro-particle enzyme immunoassay analysis and ELISA. RESULTS: In chronic HBV patients with positive HBV DNA, the positive percentage were 75% for HBeAg, 25% for anti-HBe, 69% for PreS1-Ag, and 77% for PreS2-Ag. In 75 HBeAg positive cases, there were 54 cases of PreS1-Ag positive and 60 cases of PreS2-Ag positive; while among anti-HBe positive cases, there were 14 cases of PreS1-Ag positive and 17 cases of PreS2-Ag positive. There was no significant difference of the positive percentage of PreS1-Ag and PreS2-Ag between HBeAg positive and anti-HBe positive (both P > 0.05). Under circumstance of HBeAg positive, the percentage of both PreS1-Ag and PreS2-Ag negative (8%) was significant lower than that of anti-HBe positive (28%) (P < 0.05). There were 72 cases with consistent positive or negative for both PreS1-Ag and PreS2-Ag. CONCLUSION: For hepatitis B patients with positive HBV DNA, the differences of positive percentages for HBeAg, PreS1-Ag and PreS2-Ag are moderate. The positive percentages of PreS1-Ag and PreS2-Ag do not correlate with HBeAg positive. The negative percentages of PreS1-Ag and PreS2-Ag correlate with anti-HBe positive.

[A modified dye test for Toxoplasma gondii infection].

A modified dye test with microplate was to be established to detect Toxoplasma antibodies with cell-cultured Toxoplasma gondii. Numbers of stained and unstained tachyzoites were estimated in every 100 tachyzoites in each well after dyeing with methylene blue. The dilution with 50% tachyzoites stained was used as final dilution. Better results of the microplate dye test has been received when the concentration of tachyzoites in suspension reaches 10(9)/ml with 1% sodium citrate as accessory factor.

[Study on the correlation among quantification of HBV-DNA and HBeAg, anti-HBe in hepatitis B carriers].

OBJECTIVE: To better understand the duplication of hepatitis B virus (HBV) in order to improve clinical diagnoses and treatments via quantitative measurement of HBV-DNA and comparison of correlation of HBV-DNA with HBeAg and anti-HBe. METHODS: For 883 hepatitis B patients with positive HBsAg, HBV-DNA was measured by COBAS AMPLICOR HBV MONITOR reagent and COBAS AMPLICOR quantitative PCR instrument. Microparticle enzyme immunoassay analysis (MEIA) was then carried out with fully automatic enzyme immunoassay analysis instrument made by Abbott Axsym from the U.S. to measure HBeAg and anti-HBe. Correlation was analysed by SPSS. RESULTS: (1)Positive correlation between 690 HBV-DNA positive and HBeAg positive with r= 0. 505 (P< 0.01) was found with mean values as:HBV-DNA:7.12 x 10(12) copies/ml;HBeAg:218.31 S/CO. HBV-DNA:10(4) copies/ml, HBeAg: 104 S/CO; HBV-DNA: 10(5)-10(8) copies/ml, HBeAg: 112 S/CO; HBV-DNA: 10(9)-10(15) copies/ml, HBeAg: 252 S/CO. (2) No correlation was found between 193 HBV-DNA and anti-HBe + with r= -0.052(P= 0.477> 0.05) with Mean: HBV-DNA: 8.0x 10(10) copies/ml anti-HBe: 0.18 S/CO. CONCLUSION: HBV-DNA and HBeAg appeared to have had linear correlation, showing that HBeAg> 100 S/CO,HBV-DNA> 10(4) copies/ml and hepatitis B virus were reproduced. However, HBV-DNA did not show linear correlation with anti-HBe as HBeAg negative and anti-HBe positive, the level of hepatitis B viral replication decrease slightly. But the virus load is still high. Infectivity can not neglect.

[Correlation of hepatitis B e antigen with hepatitis B virus DNA in the serum of chronic hepatitis B patients after treatment].

OBJECTIVE: To analyze the correlation of hepatitis B virus (HBV) DNA with the serological markers of HB in the serum of chronic HB patients after treatment PCR method and to analyze the status of these markers and the multiplication of virus. METHODS: Peripheral blood samples were collected from 480 chronic HB patients, aged 15 - 50, who had been treated by anti-nucleotide drugs or traditional Chinese herbs and showed normal ALT/AST. Both COBAS AMPLICOR HBV MONIORTM kit (internal-standard PCR method) and Light Cycler real time fluorescent quantitative PCR instrument (external-quantitative standard PCR method) were used to measure the HBV DNAS level. 42 of the 312 patients with the HBV DNA level lower than the minimum test limit measured by COBAS AMPLICOR HBV MONIORTM kit and HBeAg positive (>4 S/CO) underwent microparticle enzyme immunoassay (MEIA) to test the HBsAg, anti-HBs, HBeAg, anti-HBe, and HBcIgG. RESULTS: Seven of the 42 patients with HBV DNA negative measured by COBAS AMPLICOR HBV MONIORTM kit lower then the minimum test limit were shown as HBV DNA positive by Light Cycler real time fluorescent quantitative PCR. The 42 patients were HBsAg (+), anti-HBs (-), HBeAg (+), anti-HBe (-), and anti-HBcIgG (-), with an average HBeAg level of 42.26 S/CO and a positive HBeAg rate of 13.46%. CONCLUSION: HBeAg positivity does not necessarily means an active multiplication of HBV. The changes of the serological markers of HBV may be not consistent with that of HBV DNA. It is more objective to undergo both internal-standard and external-quantitative standard methods.

[Animal model of facial neuritis induced by herpes simplex virus].

OBJECTIVE: To study the role of herpes simplex virus type 1 ( HSV-1 ) in facial paralysis by developing an experimental animal model of viral facial paralysis. METHODS: Both sides of posterior auricular branch of facial nerve were anatomies and incised in 66 mice. The HSV-1 was inoculated into right ear branch and fetal bovine serum was inoculated into left ear branch as control. The symmetry of mouse face was observed and scored. The temporal bones were serially sectioned and stained with hematoxylin and eosin. The extratemporal facial nerves were stained with osmium tetroxide. HSV-1 DNA in bilateral facial nerve, brain stem, trigeminal ganglion and spinal cord was detected by the polymerase chain reaction. RESULTS: Twenty-eight (42. 42%) mice developed right facial paralysis between 2 and 5 days after inoculation. Continuing 3-6 days, the facial paralysis recovered spontaneously. Thirty-eight mice had no signs of facial paralysis. Compared with the left, nerve swelling, inflammatory cell infiltration were manifested in right temporal facial nerve of paralyzed mice. The ratio of the cross-sectional area of the facial nerve to the facial canal ( FN/FC ) was significantly higher than that on the control side (P < 0.01). Demyelinated nerve fibers were seen in the right extratemporal facial nerve. Not only in paralyzed mice, but also in non-paralyzed mice, HSV DNA was detected in some nerve tissues. CONCLUSIONS: Inoculating HSV-1 into posterior auricular branch of facial nerve can produce an acute and transient facial paralysis in mice. The possible pathophysiologic mechanism of the facial paralysis is viral invasion and transportation from distal branch to main trunk. Then the viral facial neuritis causes facial paralysis.

Role of perioperative parenteral nutrition in severely malnourished patients with Crohn's disease.

AIM: To evaluate the effect of perioperative parenteral nutrition on serum immunoglobulin, weight change, and post-operative outcome in severely malnourished patients with Crohn's disease. METHODS: Thirty-two severely malnourished patients with Crohn's disease who had undergone surgery in our hospital were reviewed. Sixteen patients who received perioperative parenteral nutrition were enrolled in the study group, and the other 16 patients who did not receive parenteral nutrition were enrolled in the control group. Serum immunoglobulin, body mass index (BMI), liver function, weight change, and postoperative complications were evaluated. RESULTS: Serum IgM levels elevated 1 wk before surgery in both groups, and decreased to normal value (from 139+/-41 to 105+/-29 mg/dL, P = 0.04) 4 wk after operation in the study group, while no significant changes was noted in the control group (from 133+/-16 to 129+/-13 mg/dL, P = 0.34). There were no significant changes in concentrations of IgG and IgA. The BMI of the study group increased from 13.9+/-0.6 to 15.3+/-0.7 kg/m(2) (P = 0.02) with no significant change in the control group (14.1+/-0.7 and 14.5+/-0.5, respectively, P = 0.81). The percentage of resuming work was higher in the study group than in the control group. CONCLUSION: Perioperative parenteral nutrition possibly ameliorates the humoral immunity, reverses malnutrition, and facilitates rehabilitation.

Screening and identification of linear B-cell epitopes and entry-blocking peptide of severe acute respiratory syndrome (SARS)-associated coronavirus using synthetic overlapping peptide library.

A 10-mer overlapping peptide library has been synthesized for screening and identification of linear B-cell epitopes of severe acute respiratory syndrome associated coronavirus (SARS-CoV), which spanned the major structural proteins of SARS-CoV. One hundred and eleven candidate peptides were positive according to the result of PEPscan, which were assembled into 22 longer peptides. Five of these peptides showed high cross-immunoreactivities (approximately 66.7 to 90.5%) to SARS convalescent patients' sera from the severest epidemic regions of the China mainland. Most interestingly, S(471-503), a peptide located at the receptor binding domain (RBD) of SARS-CoV, could specifically block the binding between the RBD and angiotensin-converting enzyme 2, resulting in the inhibition of SARS-CoV entrance into host cells in vitro. The study demonstrated that S(471-503) peptide was a potential immunoantigen for the development of peptide-based vaccine or a candidate for further drug evaluation against the SARS-CoV virus-cell fusion.

[Detection of eotaxin and its clinical diagnosis value in patients with bronchial asthma].

OBJECTIVE: To explore the role of eotaxin in the pathogenesis of bronchial asthma and the clinical value in the diagnosis of asthma. METHODS: Serum eotaxin were measured by ELISA in 38 patients with asthma, 28 patients with non-asthma allergy, and 30 healthy controls. RESULTS: The levels of serum eotaxin in the asthma group were higher than those in the non-asthma allergic and control group (P<0.01). Furthermore, eotaxin levels in patients with acute asthma were significantly higher than those in patients with stable asthma (P<0.001). It was also found that the eotaxin levels of the acute asthma group were positively correlated to the amounts of eosinophils in peripheral blood (r=0.4196, P<0.001), and inversely correlated to the forced expiratory volume in one second (FEV1) (r=-0.3746, P<0.001). CONCLUSION: It suggests that eotaxin may play a crucial pathogenic role in the asthmatic process possibly by activating the allergic inflammatory cells and controlling the recruitment of eosinophils from blood to bronchial epithelium of the airway. The concentration of eotaxin is significantly associated with the attack of acute asthma and its severity. Eotaxin may be a potential therapeutic target in patients with asthma.