Expert Consensus on Otorhinolaryngology Head and Neck Diseases Responding Specifically to Traditional Chinese Medicine / 中国实验方剂学杂志

Traditional Chinese medicine （TCM） and western medicine have their respective advantages and limitations in the diagnosis and treatment of common otorhinolaryngology head and neck diseases. Although the integrated TCM and western medicine exhibits definite curative effects， there is no consensus on the otorhinolaryngology head and neck diseases responding specifically to TCM or integrated TCM and western medicine， as well as the diagnosis and treatment schemes. The China Association of Chinese Medicine （CACM） thus organized the otorhinolaryngology head and neck specialists of both TCM and western medicine to discuss the etiology， pathogenesis， and clinical diagnosis and treatment methods of common otorhinolaryngology head and neck diseases with the results of multiple clinical trials taken into account. The acute pharyngitis， chronic pharyngolaryngitis， paraesthesia pharyngis， hysterical aphasia， allergic rhinitis， subjective tinnitus， and otogenic vertigo were confirmed to respond specifically to TCM or integrated TCM and western medicine. Then a mutually agreed diagnosis and treatment scheme and recommendation with integrated TCM and western medicine was formulated as a reference for clinical practice， thus benefiting more patients.

Mechanism of volatile oil from Alpinia oxyphylla in treating Alzheimer's disease based on GC-MS and network pharmacology / 中国中药杂志

To study the material basis and mechanism of volatile oil from Alpinia oxyphylla in treating Alzheimer's disease(AD) based on GC-MS and network pharmacology. Ingredients of volatile oil from A.oxyphylla were analyzed by GC-MS. Targets of those ingredients were obtained through Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP). Relevant targets of AD were obtained through such databases as DrugBank, STITCH, OMIM. Intersection targets of ingredients and diseases were obtained by Online Venny map, and PPI network was established by STRING to screen out core targets. Gene ontology(GO) functional enrichment analysis and KEGG pathway enrichment analysis were performed by DAVID. The "ingredients-target-pathway" network was constructed by software Cytoscape 3.8.1 to screen out potential active ingredients of volatile oil from A.oxyphylla in the treatment of AD. The results showed that a total of 6 active ingredients were screened from the volatile oil of A.oxyphylla by GC-MS, 17 targets corresponding to 6 active ingredients were found in TCMSP database, and 3 448 AD targets were found in DrugBank database. "Ingredients-target-pathway" network and PPI network showed there were 4 potential active ingredients in the treatment of AD and 4 core targets. GO analysis and KEGG analysis showed 34(P<0.05) and 5(P<0.05) pathways, respectively, including nerve ligand receptor interaction, calcium signaling pathway, cholinergic synapse and 5-hydroxytryptaminergic synapse. This suggested that volatile oil from A.oxyphylla could synergistically treat AD by regulating calcium balance, cholinergic balance and phosphorylation. This study provided reference and guidance for further study of volatile oil from A.oxyphylla in the treatment of AD.

Effect of Sijunzi Decoction on regulation of Aβ-related proteins across blood-brain barrier / 中国中药杂志

According to traditional Chinese medicine, "spleen transport" is closely related to the metabolism of substance and energy. Studies have shown that Alzheimer's disease(AD) is a disease related to glucose and lipid metabolism and energy metabolism. The traditional Chinese medicine Jiangpi Recipe can improve the learning ability and memory of AD animal model. Sijunzi Decoction originated from Taiping Huimin Hefang Prescription is the basic prescription for strengthening and nourishing the spleen, with the effects of nourishing Qi and strengthening the spleen. In this experiment, human brain microvascular endothelial cells(HBMEC) and Sijunzi Decoction water extract(0.25, 0.5, 1 mg·L~(-1)) were pre-incubated for 2 h, and then Aβ_(25-35) oligomers(final concentration 40 μmol·L~(-1)) was added for co-culture for 22 hours. The effect of Sijunzi Decoction on the activity of Aβ_(25-35) oligomer injured cells and the expression of related proteins were investigated. Q-TOF-LC-MS was used first for principal component analysis of Sijunzi Decoction water extract. Then MTT assay was used to investigate the effect of Sijunzi Decoction water extract on the proliferation of HBMEC cells. Real-time fluorescence quantitative PCR(RT-qPCR) was employed to detect the mRNA expression of GLUT1, RAGE, and LRP1. The expression of Aβ-related proteins across blood-brain barrier(RAGE, LRP1) was detected by Western blot. The results showed that 40 μmol·L~(-1) Aβ_(25-35) oligomers could induce endothelial cell damage, reduce cell survival, increase expression of RAGE mRNA and RAGE protein, and reduce expression of GLUT1 mRNA, LRP1 mRNA, and LRP1 protein. Sijunzi Decoction water extract could reduce the Aβ_(25-35) oligomer-induced cytotoxicity of HBMEC, decrease the expression of RAGE mRNA and RAGE protein, and increase the expression of GLUT1 mRNA, LRP1 mRNA and LRP1 protein. The results indicated that Sijunzi Decoction could reduce the injury of HBMEC cells induced by Aβ_(25-35) oligomer, and regulate the transport-related proteins GLUT1, RAGE and LRP1, which might be the mechanism of regulating Aβ_(25-35) transport across the blood-brain barrier.

Advances of studies on effect of drug transporter under hypoxia / 中国药理学通报

Plateau environment has the characteristic of low ox-ygen and low pressure, which leads to a series of physiological changes and affects the process of drug metabolism in the body. Many factors affect the pharmacokinetic parameters, including gastric emptying, blood rheology, cardiopulmonary function, hepatorenal function, cytochrome P450 enzyme and so on. The present study focuses on drug metabolic enzymes, since drug transporter is the key factor that mediates drugs in their entrance to the body through the cell membrane, producing the curative effect. In order to provide the reference to further research on the effect of plateau hypoxia on pharmacokinetics and guide the rational use of drugs, we review in this paper the classification of the transporter, mediated drug substrates, the influence of hypoxia on expression levels of drug transporter substrates and the regulatory mechanism of drug transporter under the condition of hypoxia.

Effect of amoxicillin on the expression of PEPT1 and pharmacokinetics upon acute hypoxia at high altitude in rat / 药学学报

The relationship between PEPT1 (peptide transporter) and drug efficacy has drawn more and more attention in the treatment of disease. PEPT1 represents a promising strategy for improvement of drug bioavailability and an important starting point for clinical rationalization of drug selection. The effect of PEPT1 on transport and pharmacokinetics of amoxicillin was investigated under hypoxia condition at high altitude in rat. The mRNA and protein expressions of PEPT1 were increased by 36.87%, 216.21%, 577.8% and 535.9% respectively in the hypoxia group in the small intestine and kidney of rats. However, the mRNA and protein expressions of PEPT1 were reduced by 43.90% and 84.7% in the liver. Compared with the control group, the AUC, tmax, Cmax, MRT and t1/2 of amoxicillin were significantly enhanced by 312.17%, 63.04%, 110.93%, 67.11% and 16.96% respectively in the hypoxia group, while the CL was significantly decreased by 74.51%. After acute exposure to high altitude, the expressions of drug transporter PEPT1 were distinctly changed in rat tissues, which can affect the pharmacokinetics of amoxicillin.

Effects of China diabets educator training and its influencing factors / 中华护理杂志

Objective To describe the effects of China diabetes educator training and explore the influencing factors.Methods Totally 314 diabetes educators who graduated from Johnson & Johnson Diabetes Institute(JJDI) from 2007 to 2014 were recruited and investigated via self-designed evaluation form.Results The total score was 76.64±22.24.The top three items with highest scores were performing nurses training (93.36±62.81),form of health education (84.27±23.09) and working responsibility and procedure of diabetes educator (82.07±32.43).The items scored lower than 60 were regular follow-up system(56.16±31.74),publishing health education papers(38.47±47.75) and conducting related researches (26.11±44.00).Multiple linear regression analysis showed that educators who had longer time of working in diabetes department(t=3.515,P=0.001) and whose post were educators(t=3.404,P=0.001) and who were not first line practitioners (t=-2.589,P=0.001) had higher scores.Conclusion The educators' main work in China is focusing on regular health education.Follow-up management and research renovation capacity should be enhanced in the future for educators.

Evaluation of the effect of chronic virus infection on laboratory tests results in patients with osteoarticular tuberculosis / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To evaluate the effect of chronic virus infection on laboratory tests results in patients with osteoarticular tuberculosis.</p><p><b>METHODS</b>A total of 121 patients with osteoarticular tuberculosis, who were hospitalized in Shenzhen Third People's Hospital during June 2008 to June 2012, were recruited for analysis. Clinical laboratory tests results were collected for comparison between patients with or without chronic co-infection with virus.</p><p><b>RESULTS</b>Among the 121 patients, thirty patients were co-infected with hepatitis B virus (HBV), two were with Human immunodeficiency virus (HIV), and one was co-infected with HBV, HIV and hepatitis C virus (HCV). Compared to patients with osteoarticular tuberculosis without HBV/HCV/HIV infection, patients with chronic HBV/HCV/HIV virus infection had similar positive rate of laboratory tests including tissue smear acid-fast bacilli (AFB) staining, tissue Mycobacterium tuberculosis (Mtb) culture, tissue Mtb DNA detection, serological test of antibodies against Mtb, and Mtb. antigen-specific interferon-gamma release assay. Similar results were also found for erythrocyte sedimentation rate, C-reative protein level and liver function including Alanine aminotransferase and Aspartate Aminotransferase.</p><p><b>CONCLUSION</b>Chronic infection with HBV/HCV in patients with have no obvious effect on clinical laboratory tests related to tuberculosis.</p>

The genetical evolution of the full length genes of 5 EV 71 strains from 5 Shenzhen patients with hand-food-mouth disease associated with EV71 infection / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate the characteristics of molecular epidemiology and molecular evolution of 5 EV 71 (enterovirus 71, EV71) strains from 5 Shenzhen patients with hand-food-mouth disease associated with EV 71 infection.</p><p><b>METHODS</b>5 EV 71 strains were isolated, and sequenced to analyzed the full length gene sequences in order to compare nucleotide and amino acid homology with other EV71 strains from other regions and countries as well as previous strains across the world through bioinformatics software.</p><p><b>RESULTS</b>5 strains of EV 71 belonged to sub-genotype C4 by analysis of nucleotide sequences of VP1 and VP4 of EV 71. The differences of nucleotide and amino acid sequences were much small with nucleotide homology of 93% and amino acid homology of 98% among these 5 strains. A phylogenetic tree analysis indicated that 2008 Shenzhen epidemic strains were the most close to 2004 Shenzhen circulating strains, and also much close to 1998 Shenzhen epidemic strains and 2008 Fuyang Anhui strains. The dead strain was very close to 2008 Fuyang Anhui epidemic strains.</p><p><b>CONCLUSION</b>It can be speculated that this epidemic strains of EV 71 probably originate from the same ancient strain in the history, may from 1998 Shenzhen strain.</p>

Loss of the balance between Th17 and Th1 populations in HIV/AIDS patients / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To evaluate the Th17/Th1 response in HIV infected patients and the mutual relationship between the response of Th17 and Th1.</p><p><b>METHODS</b>38 chronic HIV infected patients as well as 24 healthy volunteers were performed in this study. The patients were divided into two groups, one group before treatment, the other after therapy. The whole blood intracellular cytokine staining was used, samples detected by BD FACSCanto, after that, the expression of CD4+ IL-17+ T cell and CD4 IFN-gamma+ T cell were analyzed by FACSDiva software and lastly compared the differences among different groups.</p><p><b>RESULTS</b>The expression of CD4+ IL-17+ T cell in naive-therapy patients were significantly lower than that of the healthy controls (1.14 +/- 0.7)9% vs (3.98 +/- 1.14)%, P = 0.000, but increased remarkably after HARRT(highly antiretroviral treatment) (2.22 +/- 1.00)%, P = 0.001; however there were no significant differences in the expression of CD4+ IFN-gamma+ T cell before and after therapy (34.35 +/- 24.38)% vs (42.10 +/- 15.57%), also with the healthy control (P = 0.383). The frequency of CD4 IL-17+ T cell was positively correlated with CD4+ T counts (R = 0.345, P = 0.034), but no significant correlations was observed between the expression of CD4+ IFN-gamma T cell and CD4+ T counts (R = -0.247, P = 0.136).</p><p><b>CONCLUSION</b>The infection of HIV virus down-regulated Th17 immune response and disturbed the balances between Th17 and Th1 evidently in human. Th17 response may play an important role in the pathogenesis of HIV infection.</p>

Study on characteristics of cellular-mediated immune responses of novel H1N1 influenza A patients with pneumonia / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate the phenotype, frequency and function of CD4+ T cell subsets and the relevant cytokines, as well as the relationship between these cells and appearance of pneumonia of novel (H1N1) influenza A patients.</p><p><b>METHODS</b>68 healthy people, 53 confirmed novel A(H1N1) influenza patients without pneumonia and 16 confirmed severe novel A (H1N1) influenza patients with pneumonia were enrolled in this study. Viral load in nasopharyngeal swabs specimens was measured by real time PCR assay. The phenotype and percentage of CD4+ T cell subsets including Th1, Th2, Th17, and Treg cells were measured by Flow cytometry analysis. The relevant cytokines in plasma including TGF-beta, IL-6 and IFN-gamma were measured by ELISA. Data was analyzed by one way ANOVA.</p><p><b>RESULTS</b>It was found that peak viral load and viral shedding period of severe patients with pneumonia was significantly increased compared with mild patients without pneumonia (P < 0.05). The percentage of Th17 cells of severe patients with pneumonia was significantly diminished compared to that of healthy subjects and mild patients without pneumonia (P < 0.05). However, Th1, Th2, Treg cells frequencies had no significant differences (P > 0.05) among these three groups. The level of TGF-beta in plasma for the severe patients with pneumonia was also significantly decreased compared to that of healthy subject and mild patients without pneumonia (P < 0.05). The viral shedding period inversely correlated with the frequency of Th17 cells (r = - 0.38, P < 0.05).</p><p><b>CONCLUSION</b>H1N1 influenza A virus can inhibit Th17 cells to differentiate, particularly more extent in patients with pneumonia. Impaired Th17 cells may correlate with viral clearance and pneumonia of novel H1N1 influenza A patients.</p>

Preparation of human anti-HBs from volunteers with hepatitis B vaccine boost vaccination by modified B-cell immortalization technique / 南方医科大学学报

<p><b>OBJECTIVE</b>To establish immortalized B lymphoblast cell lines (B-LCLs) from healthy anti-HBs antibody (anti-HBs)-positive volunteers and screen for human anti-HBs and the antibody-secreting cells.</p><p><b>METHODS</b>The peripheral blood mononuclear cells (PBMC) isolated from 3 healthy volunteers positive for anti-HBs with hepatitis B vaccine boost vaccination were infected with Epstein-Barr virus (EBV) and incubated in the presence of CpG DNA motifs and cyclosporin A (CyA). The anti-HBs in the culture supernatant of the immortalized B-cells was quantified by Architect anti-HBs assay with chemiluminescent microparticle technique. Immunocytochemistry was performed to identify the differentiation of the cell clones expressing anti-HBs.</p><p><b>RESULTS</b>Immortalized B-cell culture was successfully established from the cell clones secreting anti-HBs with EBV infection and CpG DNA stimulation. The titer of anti-HBs in the culture supernatant was at its peak at 3 weeks of cell culture and then decreased gradually. At 3 months of cell culture, the cells still retained the capacity of anti-HBs production as verified by the results of immunocytochemistry for CD20 and CD138.</p><p><b>CONCLUSION</b>Immortalized B-cell culture secreting anti-HBs from volunteers receiving boost hepatitis B vaccination has been successfully established by modified EBV immortalization technique.</p>

The association between HBV genotyping and clinical characteristics and expression of TH1/TH2 cytokines / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To explore the association between HBV genotyping and clinical characteristics and expression of TH1/TH2 cytokines.</p><p><b>METHODS</b>The expression of IL-4 and IFN-gamma was detected with flow cytometry for 102 HBV infections and 48 healthy controls. 50 CHB patients were randomly selected for HBV genotyping with real-time fluorescence PCR assay.</p><p><b>RESULTS</b>Higher expression of IL-4 in peripheral blood was detected in patients with HBV infection than healthy controls (P < 0.001); No significant differences on expression of Th1/Th2 cytokines were observed in CHB patients with different HBV DNA levels or HBeAg status (P > 0.05). There were 34 (68%) patients with genotype B infection and 16 (32%) with genotype C infection. Compared to patients with genotype B infection, the patients with genotype C infection showed higher levels of IL-4 (P = 0.018), and Th1/Th2 ratio decreased,but the difference was not statistically significant (P = 0.2262).</p><p><b>CONCLUSION</b>The different expression of TH1/TH2 cytokines may elucidate cellular immune response and clinical outcome difference between patients with genotype B infection and genotype C infection.</p>

The expression of some homing and co-receptor molecules on CD4+ T cells in AIDS patients / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To evaluate the expression of CD49d, CCR9, CD62L and CCR5 on CD4+ T cells in AIDS patients before and after HAART.</p><p><b>METHODS</b>The study was performed in 42 cases of AIDS patients and 18 cases of healthy controls. The expression of CD49d, CCR9, CD62L and CCR5 on CD4+ T cells, and CD45RO on CD4+ CCCR9+ and CD4+ CCR5+ T cells in AIDS patients and healthy controls were analysed by Flow cytometry. Software BD FACSDiva was used to calculate the percentage of expression.</p><p><b>RESULTS</b>The number of peripheral CD4+ T cells in group pre-HAART was decreased compared with group HAART (P < 0.01); the frequency of CD3+ CD4+, CD4+ CCR9+, CD4+ CCR5+ T cells in group pre-HAART were decreased compared with group HAART (P < 0.01), the frequency of CD4+ CD49d+, CD4+ CD62L+, CD4+ CCR9+ CD45RO , CD4+ CCR9+ CD45RO- ,CD4+ CCR5+ CD45RO+, CD4+ CCR5+ CD45RO- T cells in group pre-HAART were significantly decreased compared with group HAART (P < 0.001); the frequency of CD3+ CD4, CD4+ CD62L+, CD4+ CCR5+ T cells in group HAART were significantly decreased compared with group HIV-neg (P < 0.05).</p><p><b>CONCLUSIONS</b>Not only the number but the function of CD4' T cells was impaired in AIDS patients: the lower expression frequency of gut homing receptor molecule CD49d and CCR9,1ymph node homing molecule CD62L, coreceptor molecule CCR5. HAART can partially reverse this pathological phenomena. CD49d, CCR9 and CD62L may be suggested to indicate the progression of AIDS and immunologic reconstitution after HAART.</p>

Hepatitis B virus P22(e) inhibit hepatocyte apoptosis via nuclear factor kappa B / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To test whether nuclear factor kappa B plays an important role in the apoptosis-inhibitory effect of hepatitis B virus (HBV) P22(e) protein.</p><p><b>METHODS</b>HepG2 cells were transfected with recombination plasmid pEGFP-HBVP22(e). The Act-D and TNF alpha were used to induce apoptosis. NF-kappa B inhibitor ALLN were used to inhibit the signaling pathway. The activation of NF-kappa B was EMSA, and the nulear translocation of NF-kappa B was determined by immuno-staining.</p><p><b>RESULTS</b>Laser scanning confocal microscopy and EMSA indicated that HBV P22(e) protein enhanced the nuclear translocation of NF-kappa B after apoptosis induction. ALLN treatment inhibited the nuclear translocation of NF-kappa B, and blocked the apoptosis-inhibiting effect of HBV P22(e) protein.</p><p><b>CONCLUSION</b>This study indicates that HBV P22(e) protein inhibits apoptosis of hepatocyte via the NF-kappa B signaling pathway.</p>

Study of the clinical and laboratory features of hand-foot-mouth disease / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To study the clinical and laboratory features of the mild and severe hand-foot-mouth diseases (HFMD) in Shenzhen in 2008.</p><p><b>METHODS</b>145 cases were observed in East-Lake Hospital and Shenzhen Children's Hospital. Of the 145 cases, 124 mild cases and 21 severe cases were involved.All the clinical data and laboratory findings were collected and summarized. After collection of the acute and convalescent consecutive stools and peripheral bloods from the patients with HFMDI, EV71 genes were amplified from these samples by RT-PCR. Enterovirus 71 were cultured and isolated using Vero cell line and R&D cell line.</p><p><b>RESULTS</b>The WBC counts and blood glucose levels of the severe cases were significantly elevated, but the ages of the severe ones significantly decreased compared with those of the mild cases (P < 0.05). EV71 genes could be detected by RT-PCR with 35% positive rate in mild cases and 67% in severe cases. The EV71 gene detection rate of the severe cases was significantly increased in contrast to that of the mild ones. The EV71 were isolated and cultured from the stools of 9 patients, one specimens from the dead's stool. Two severe cases died of neurogenic pulmonary edema and brain-stem encephalitis.</p><p><b>CONCLUSIONS</b>EV71 mainly contributes to HFMD and is responsible for death of some severe cases. High fever, less rash, elevated white blood cell counts and blood glucose concentrations as well as age less than 4 years old should be used for prediction of severe cases.</p>

The influence of HBV/P22 protein on the apoptosis of HepG2 cells: an experimental study / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To study the influence of HBV/P22 protein on the induced apoptosis of HepG2 cells.</p><p><b>METHODS</b>In vitro, two HepG2 strains were transfected with pcDNA3.1+ and pcDNA3.1+HBV/P22 respectively and the cells were exposed to Act D and TNF alpha for 6h and then the induced apoptosis was detected by flow cytometry (FCM) and TUNEL technique. Supernatant HBeAg was detected by Abbott reagent. The intracellular expression of HBV/P22 protein was measured by Western blot and immunochemistry. In vivo, three cell groups were inoculated into nude mice by subcutaneous injections. After two weeks, Act D and TNF alpha were injected into the tumors and then the induced apoptosis in the tissues was detected by TUNEL technique. The expression of HBV/P22 protein in the tumor tissues was detected by immunochemistry.</p><p><b>RESULTS</b>In vitro, in HepG2- pcDNA3.1+HBV/P22 cells, supernatant HBeAg was positive and intracellular HBV/P22 protein was positively expressed. The apoptosis proportion of HepG2-pCDNA3.1+HBV/P22 cells was markedly lower than HepG2 and HepG2-pcDNA3.1+ cells (P < 0.05). In vivo, HBV/P22 protein was expressed in the tumor tissues, and the apoptosis proportion in the group injected with HepG2-pcDNA3.1+HBV/P22 cells was markedly lower than those injected with HepG2 and HepG2-pcDNA3.1+cells (P < 0.05).</p><p><b>CONCLUSION</b>HBV/P22 protein could inhibit the induced apoptosis of HepG2 cells both in vitro and in vivo.</p>

Hepatitis B virus P22e protein inhibits human hepatocellular carcinoma HepG2 cell apoptosis in vitro / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effects of the hepatitis B virus (HBV) P22e protein on the apoptosis of human hepatocellular carcinoma HepG2 cells.</p><p><b>METHODS</b>HepG2 cells were transfected with recombinant plasmid pEGFP-HBVP22e and exposed to Act-D and tumor necrosis factor alpha (TNFalpha) treatment to induce cell apoptosis. Flow cytometry was performed to determine the proportion of cells containing sub-G1 DNA to represent the number of apoptotic cells. Laser scanning confocal microscopy was used to observe the nuclear alterations in the apoptotic cells.</p><p><b>RESULTS</b>HepG2EGFP-C2HBVP22e cell strain showed a much delayed apoptosis as well as obviously lowered apoptotic rate in comparison with the HepG2 strain (P<0.01).</p><p><b>CONCLUSION</b>The introduction and expression of extraneous gene HBVP22e significantly inhibits the apoptosis of HepG2 cells.</p>

Viral breakthrough and neutralizing anti-interferon antibody production in chronic hepatitis B patients treated with recombinant interferon-alpha / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the relationship of virological breakthrough and production of neutralizing anti-interferon antibody (NAb) in chronic hepatitis B patients treated with recombinant interferon-alpha (rIFN-alpha).</p><p><b>METHOD</b>Four hundred eighty-five patients with histological proven chronic hepatitis B were treated with 5 MU recombinant interferon-alpha 1b (rIFN-alpha1b) thrice weekly for 6-37 months (median 10). Serum HBV DNA, HBeAg and NAb levels of the patients were detected by fluorescent-quantitative PCR, enzymoimmunoassay and antiviral neutralizing biological assay respectively during the therapy.</p><p><b>RESULTS</b>Virological breakthrough occurred in 66 patients (13.6%), and NAb was found in 98 patients (20.2%) of the total 485 patients. The rate of NAb positivity was higher in patients with viral breakthrough than those without it (68.2%, 45/66, vs 12.6%, 53/419, chi(2)=109.06, P < 0.01), and viral breakthrough occurred more in patients with positive NAb than with negative NAb (45.9%, 45/98, vs 5.4%, 21/387, chi(2)=109.06, P < 0.01). The time of the viral breakthrough occurrence and the time of NAb production had a significant correlation (P < 0.01). The occurrence of viral breakthrough was also influenced by the age of patients (P < 0.05) and HBeAg status (P < 0.01) before they were treated.</p><p><b>CONCLUSION</b>Viral breakthrough occurred in 13.6% of our 485 chronic hepatitis B patients treated with recombinant interferon-alpha. Their viral breakthrough and production of NAb production had a significant correlation.</p>

Stable expression of HBV C gene mutants in immortalized human B-cell lines / 南方医科大学学报

<p><b>OBJECTIVE</b>To provide an cell model of immortalized lymphoblstoid B-cell lines for studying the biological characteristics of full-length hepatitis B virus (HBV) genome carrying the hot-spot mutations V60, G87, and L97.</p><p><b>METHODS</b>V60, G87, and L97 mutation points were introduced into HBV p3.8 II plasmid containing 1.2 copy of HBV genome by means of site-directed mutagenesis. The HBV genome was amplified by PCR from p3.8 II and p3.8 II-V60, G87, L97 plasmid, and the PCR product was inserted into EBO-plpp eukaryotic expression vector. The recombinant vectors and the EBO-plpp vector were transfected into immortalized human lymphoblasts with lipofectamine 2000 and selected with hygromycin. Steady expression of the target genes was determined by RT-PCR, Western blotting and microparticle enzyme immunoassay.</p><p><b>RESULTS</b>DNA sequence analysis indicated that the desired mutation was introduced into wild-type HBV DNA. HBsAg, HBeAg and HBcAg could be detected in EBO-HBV-transfected cell lysate or culture supernatant.</p><p><b>CONCLUSION</b>Transfectants that stably express HBV mutant antigen may provide a cell model to study the biological characteristics of HBV carrying hot-spot mutation in vitro.</p>

Correlation between Serum Interleukin -4,12 and Cellular Immunity in Children with Asthma / 实用儿科临床杂志

Objective To study the changes of serum interleukin(IL)-4,IL-12 and correlation with cellular immunity in children with asthma of different stages.Methods Fifty asthmatic children were randomly selected, including 30 cases in attack stage (group A) and 20 cases in remission stage (group R). At the same time, 22 healthy children were studied as normal controls (group N).The levels of IL-12 and IL-4 ,T cells subgroups and erythrocyte immunity were detected.Results 1.Serum IL-12 levels were (24.44? 13.26 ),(42.30?12.65),(44.68?28.28) ng/L in group A, R and N,respectively. There was significant difference in three groups (F=8.92 P