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Biomedical subjects

Li Huang

Publications and source records attributed to Li Huang.

At least 37 records · Page 2Linked to original sources

Assessment and prognostic significance of mitotic index using the mitosis marker phospho-histone H3 in low and intermediate-grade infiltrating astrocytomas.

Distinguishing between grade II and grade III diffuse astrocytomas is important both for prognosis and for treatment decision-making. However, current methods for distinguishing between grades based on proliferative potential are suboptimal, making identification of clear cutoffs difficult. In this study, we compared the results from immunohistochemical staining for phospho-histone H3 (pHH3), a specific marker of cells undergoing mitosis, with standard mitotic counts (number of mitoses/10 high-power fields) and MIB-1 labeling index values for assessing proliferative activity. We tested the relationship between pHH3 staining and tumor grade and prognosis in a retrospective series of grade II and III infiltrating astrocytomas from a single institution. The pHH3 index (per 1000 cells), MIB-1 index (per 1000 cells), and number of mitoses per 10 high-power fields were determined for each of 103 cases of grade II and III diffuse astrocytomas from patients with clinical follow-up. pHH3 staining was found to be a simple and reliable method for identifying mitotic figures, allowing a true mitotic index to be determined. The pHH3 mitotic index was significantly associated both with the standard mitotic count and with the MIB-1 index. Univariate analyses revealed that all 3 measurements of proliferation were significantly associated with survival. However, the pHH3 mitotic index accounted for a larger proportion of variability in survival than standard mitotic count or MIB-1/Ki-67 labeling index. After adjusting for age, extent of resection, and performance score, the pHH3 mitotic index remained an independent predictor of survival. Thus, pHH3 staining provides a simple and reliable method for quantifying proliferative potential and for the stratification of patients with diffuse astrocytomas into typical grade II and III groups. These results also suggest that pHH3 staining may be a useful method in other neoplasms in which accurate determination of proliferation potential is relevant to tumor grading or clinical treatment decision-making.

Adult↗

Oligonucleotide cleavage and rejoining by topoisomerase III from the hyperthermophilic archaeon Sulfolobus solfataricus: temperature dependence and strand annealing-promoted DNA religation.

Topoisomerase III from the hyperthermophilic archaeon Sulfolobus solfataricus (Sso topo III) is optimally active in DNA relaxation at 75 degrees C. We report here that Sso topo III-catalysed DNA cleavage and religation differed significantly in temperature dependence: the enzyme was most active in cleaving ssDNA containing a cleavage site at 25-50 degrees C, but was efficient in rejoining the cleaved DNA strand only at higher temperatures (e.g. > or = 45 degrees C). The failure of Sso topo III to rejoin the cleaved DNA strand efficiently appeared to be responsible for the inability of the enzyme to relax negatively supercoiled DNA at low temperature (e.g. 25 degrees C). Intriguingly, Sso topo III facilitated DNA annealing although it showed higher affinity for ssDNA than for dsDNA. Religation of the DNA strand cleaved by Sso topo III was drastically enhanced when the DNA was allowed to anneal to a complementary non-cleaved oligonucleotide, presumably as a result of destabilization of the interaction between the enzyme and the cleaved strand through the formation of duplex DNA. A region in the non-cleaved strand corresponding to a sequence containing six bases on the 5' side and two bases on the 3' side of the cleavage site in the cleaved strand was crucial to the annealing-promoted religation. However, the annealing-promoted religation was relatively insensitive to mismatches in this region and the region conserved for oligonucleotide cleavage, except for that at the 5' end of the broken strand. These results suggest that Sso topo III is well suited for a role in DNA rewinding, whether it leads to homoduplex or heteroduplex formation.

Archaeal Proteins↗

Melatonin reduces infarction volume in a photothrombotic stroke model in the wild-type but not cyclooxygenase-1-gene knockout mice.

Cyclooxygenase (COX)-2 plays a harmful role in cerebral ischemic/reperfusion injury, but the role of COX-1 is uncertain. In the present study, cerebral infarct was induced by photothrombosis. Intraperitoneal injections of melatonin at 15 g/kg or its vehicle were made at 0.5 hr before stroke and 24 and 48 hr after stroke. Cerebral blood flow (CBF) in the penumbra was monitored during stroke using a laser Doppler flowmeter. Sensorimotor behavior was evaluated using the turning in an alley and falling from a pole tests at 1 hr before stroke and 24 and 48 hr after stroke. Infarct volume was determined from the T2-weighted magnetic resonance images at 72 hr after stroke. During the first 15 min of stroke, CBF decreased in the penumbra in both homozygous COX-1-gene knockout and wild-type mice. Melatonin treatment improved the penumbral CBF in the wild-type mice. Mild poststroke impairment in sensorimotor behavior was detected by the turning in an alley test in which the COX-1-gene knockout mice performed better. Melatonin treatment did not affect the poststroke sensorimotor behavior. The relative infarct volume at 72 hr after stroke was 8.1% and 8.4% in the COX-1-gene knockout and wild-type mice, respectively. Melatonin treatment reduced the relative infarct volume to 6.3% in the latter but not in the former (8.2%). Thus, COX-1-gene knockout does not affect the brain's susceptibility to photothrombotic stroke. Melatonin treatment reduces infarct size in the wild-type mice following photothrombotic stroke partly via maintenance of penumbral CBF in which the COX-1-gene may play a role.

Animals↗

Acute myeloid leukemia associated with variant t(8;21) detected by conventional cytogenetic and molecular studies: a report of four cases and review of the literature.

Acute myeloid leukemia (AML) with the t(8;21) (q22;q22) creating the AML1-ETO fusion gene is a distinct type of AML generally associated with a favorable prognosis. The clinicopathologic features of AML carrying variant t(8;21) are less well characterized. We report 4 cases of AML characterized by ins(8;21)(q22;q22q22), t(1;21;8)(q25;q22;q22), t(8;11;21)(q22;q13;q22), and t(4;21;8;12)(q31.3;q22;q22;q15), respectively. Fluorescence in situ hybridization or reverse transcriptase-polymerase chain reaction assay confirmed the presence of AML1-ETO or its transcripts in 3 cases assessed. Each neoplasm had morphologic characteristics of AML associated with the t(8;21). The blasts in 2 cases expressed CD19. All patients were treated with combination chemotherapy and are in complete remission, despite 2 relapses in 1 patient, with a follow-up period ranging from 8 to 46 months. We conclude that cases of AML with variant t(8;21) display morphologic, immunophenotypic, and clinical features similar to classical cases. A combination of conventional cytogenetic, and molecular analyses allows identification of these variants.

Adolescent↗

[Effects of acupuncture on the cortical functional areas activated by index finger motion in the patient with ischemic stroke].

OBJECTIVE: To observe the effect of acupuncture on cortical functional areas of the patient with ischemic stroke activated by the index finger motion. METHODS: The cortical magnetic resonance imaging (fMRI) were carried out in 15 cases of ischemic stroke during the index finger motion at acupuncture or non-acupuncture. The distribution of the cortical functional areas activated and the size of the activated region and the intension of signals were measured. RESULTS: The finger motion with no acupuncture could activate the contralateral primary somatomotor area (M1), contralateral premotor area (PMA) and contralateral first somatosensory area (S1). The finger motion with acupuncture could activate the same areas and also activate ipsilateral M1, focus area contralateral superior parietal lobule, contralateral superior temporal gyrus, and contralateral insular lobe, etc.. Both the area of the activated region and the minimum signal in the finger motion with acupuncture were statistically significantly larger than those in finger motion with no acupuncture. CONCLUSION: Rehabilitation of motor functions of the patient with ischemic stroke by acupuncture is related with improvement of blood circulation functional area in the cortex.

Acupuncture Therapy↗

[Effects of costimulatory pathway OX40/OX40L on the pathogenesis of allergic asthma in mice].

OBJECTIVE: Allergic asthma is thought to be mediated by CD4+ T lymphocytes producing the Th2-associated cytokines, which play a critical role in the development of the airway hyper-responsiveness and the eosinophilic inflammatory response. The costimulatory pathway CD28/B7 has been shown to play an important role in CD4+ T cell activation in allergic asthma. This study was conducted to evaluate the role of another costimulatory pathway OX40/OX40 ligand (L) in the pathogenesis of allergic asthma in BALB/c mice. METHODS: An allergic asthma model in BALB/c mice was established. Thirty-six BALB/c mice were randomly divided into three groups with 12 in each. Mice in treatment group (group B) were treated with neutralizing anti-OX40L monoclonal antibody (mAb, 300 microg per mouse) during the sensitization period. Mice in two control groups, asthma model group (group A) and IgG antibody group (group C) were treated with normal saline (NS) and control IgG respectively instead of anti-OX40L mAb. Bronchoalveolar lavage fluid (BALF) was collected from the mice of each group for counting the total number of white blood cells (including neutrophil granulocyte, lymphocyte, monocyte and eosinophil granulocyte) and the proportions of these cells. The levels of IL-4 and INF-gamma in BALF were measured by ELISA. Lungs were removed for morphological examination after HE and PAS staining, and expression of OX40 in lungs was evaluated by immunohistochemical method. RESULTS: (1) The count of total number of white blood cells in BALF (x10(6)/ml) was lower in group B than that of group A and group C (26.6 +/- 4.6 vs. 36.8 +/- 5.2 and 34.3 +/- 6.9, respectively), the difference between the treatment group (group B) and two control groups (groups A and C) was significant; The proportions of eosinophils and lymphocytes in the BALF (%) were lower in group B than those in group A and group C (eosinophils 15.1 +/- 2.6 vs. 20.0 +/- 4.1 and 19.9 +/- 3.9, respectively; lymphocytes 7.0 +/- 0.9 vs. 8.9 +/- 1.6 and 8.6 +/- 1.8, respectively), the difference between the treatment group and two control groups was significant. (2) The IL-4 level in BALF (pg/ml) was lower in group B than that in group A and group C (672 +/- 58 vs. 809.57 +/- 106.00 and 784 +/- 58, respectively), but the INF-gamma levels in BALF (pg/ml) were higher than those in group A and group C (0.86 +/- 0.09 vs. 0.69 +/- 0.15 and 0.67 +/- 0.13 respectively), and all the differences were statistically significant. (3) The expression of OX40 in the lungs of mice in group B were at a lower level than that of group A and group C, and the morphological changes of asthma were ameliorated in the mice of the treatment group. The signs of mice in treatment group were obviously ameliorated as compared to the two control groups. CONCLUSION: Blocking the costimulatory pathway by administering the neutralizing anti-OX40L mAb during the sensitization period of allergic asthma model could balance the Th1/Th2 responses, inhibit lung inflammation and ameliorate the signs of mice model of asthma.

Animals↗

[Construction and analysis of a metagenomic library from Tengchong hot spring soil in Yunnan Province].

By a combination of freezing/thawing/ proteinase K-based method and SDS/high-salt/heating treatment, the mixed environmental genomic DNA was isolated directly from a hot spring soil in Tengchong, Yunnan, China. With this method, The DNA yield was up to 1 - 2 microg/g soil. After purification with the Wizard DNA clean up system (Promega, Madison, Wis), the mixed genomic DNA was partially digested with restriction enzyme Pst I. Digested DNA fragments of 3 - 8 kb were recovered from agrose gel and ligated to the pSK (+) vector. The ligation mixture was transformed into DH10B strain, resulting in the construction of a metagenomic library with about 2.5 x 10(4) clones. Restriction enzyme analysis revealed that the average insert is about 4.6 kb. Some novel sequences were identified via sequencing and gene annotation analysis of 30 clones randomly chosen from this library.

China↗

[Serum HBsAg concentration and HBV replication level in hepatitis B patients with positive serum HBsAg and HBeAg].

OBJECTIVE: To analyze the relationship between serum HBsAg concentration and HBV replication level in hepatitis B patients with positive serum HBsAg and HBeAg, and to explore the possibility of using serum HBsAg concentration as a marker of HBV replication level in hepatitis B patients with positive serum HBeAg. METHODS: HBV DNA level and serum HBeAg, HBsAg concentration of 296 patients with positive serum HBsAg and HBeAg were quantitatively detected by real-time fluorescence quantitative PCR (FQ-PCR) and time-resolved fluoroimmunoassay (TRIFA) respectively. HBsAg concentrations were compared among patients with different HBV DNA levels, and HBV DNA levels were compared among patients with different HBsAg concentrations. The correlation between serum HBsAg concentration and DNA replication level were analyzed. The positive, negative predictive values and coincidence rates were speculated by various HBsAg concentrations. RESULTS: If HBV DNA positive was defined as HBV DNA levels no less than 10(5) copy/mL, then 228(77.03%) patients were classified as HBV DNA positive. HBsAg concentration was positively correlated with HBV DNA replication level, but among groups with various DNA replication levels, HBsAg concentration showed no significant statistical difference (P>0.05). If the patients were divided into 2 groups, HBsAg concentration (180 microg/L) was served as the cutoff level, the DNA positive rate of the group with HBsAg concentration no less than 180 microg/L was significantly higher than that with HBsAg concentration less than 180 microg/L (chi(2)=3.998, P<0.05). DNA positive rates and average DNA levels showed no significant statistical differences between the 2 groups, if HBsAg concentrations other than 180 microg/L were used as the cutoff level. Positive predictive values, negative predictive values and the coincidence rates speculated by various HBsAg concentrations as cutoff values did not show any significant statistical difference in estimating HBV replication levels. CONCLUSION: To some extent, serum HBsAg concentration is related to HBV DNA replication level in hepatitis B patients with positive serum HBsAg and HBeAg, but it is not feasible to use HBsAg concentration to monitor their HBV replication levels.

DNA, Viral↗

[Photocatalytic degradation of hexafluorobenzene by tungstophosphoric acid].

We verified that C6F6, a typical perfluorine compound, could be photocatalytic degraded by H3PW12O40. The defluorination of C6F6 was determined as a function of irradiation time, pH, initial concentration of H3PW12O40, and additives as well. The defluorination of C6F6 increased with the irradiation time. Acid condition, especially pH = 1.0 is preferred. 3 mL solution of 2.0 x 10(-4) mol/L C6F6 and 5.0 x 10(-6) mol/L H3PW12O40 was irradiated for 20 min and the defluorination of C6F6 was equal to 208.1%. O2, KMnO4 and K2S2O8 added in the solution were able to increase the defluorination of C6F6 via oxidating [PW12O40](4-). The defluorination of C6F6 would decline in the presence of alcohol and aromatic compounds.

Catalysis↗

[Microstructural characterization for pyrolysis hydrogenation fragments in polyethylene by pyrolysis hydrogenation gas chromatography/mass spectrometry].

The microstructural characterization of 47 samples, respectively of high density polyethylene (HDPE), low density polyethylene (LDPE), linear low density polyethylene (LLDPE), copolymer of ethylene and alpha-olefins, was performed by pyrolysis hydrogenation gas chromatography/mass spectrometry (Py-H-GC/MS) with 10% Pt on 80-100 mesh Diasolid H as a hydrogenation catalyst. The samples were obtained from 12 domestic plants and 12 plants abroad as well. The sample sizes ranging from 30 to 80 microg were pyrolyzed at 770 degrees C in a flow of hydrogen carrier gas. It was found that short methyl branches as the characteristic isoalkanes of pyrolysis hydrogenation in polyethylene would be related closely to the different classes of polyethylene. The microstructural characterization has been carried out by means of Py-H-GC/MS, according to the relative amount of the four short methyl branches, 5-methyldodecane (5-MC12), 4-methyldodecane (4-MC12), 2-methyldodecane (2-MC12) and 3-methyldodecane (3-MC12) existed in C12-C13 isoalkanes. The Py-H-GC/MS technique will be providing further scientific evidences for the characterization of polyethylene plastics in forensic science.

English Abstract↗

Synthesis and anti-HIV activity of bi-functional betulinic acid derivatives.

Betulinic acid (BA) derivatives with a side chain at C-3 can inhibit HIV-1 maturation. On the other hand, BA derivatives with a side chain at C-28 can block HIV-1 entry. In order to combine the anti-maturation and anti-entry activities in a single molecule, new bi-functional BA derivatives containing side chains at C-3 and C-28 have been synthesized. The most potent compound ([[N-[3beta-O-(3',3'-dimethylsuccinyl)-lup-20(29)-en-28-oyl]-7-aminoheptyl]-carbamoyl]methane) inhibited HIV-1 at an EC50 of 0.0026 microM and was at least 20 times more potent than either the anti-maturation lead compound DSB or the anti-entry lead compound IC9564. This bi-functional BA derivative was active against both HIV entry and maturation. These results suggest that bi-functional BA derivatives with dual mechanisms of action have the potential to become clinically useful for AIDS therapy.

Animals↗

The early embryo loss caused by 2,3,7,8-tetrachlorodibenzo-p-dioxin may be related to the accumulation of this compound in the uterus.

2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is a reproductive and developmental toxicant that can alter endocrine status, leading to decreased fertility and altered embryonic development; however, there are limited reports on TCDD toxicity during early pregnancy. In the present study, pregnant and pseudopregnant NIH mice were exposed to TCDD orally (2, 50 and 100 ng/kg body weight) during early gestation (days 1-8), pre-implantation stages (days 1-3), and peri-implantation to early post-implantation stages (days 4-8). TCDD concentration in uterus, liver, kidney, brain and fat on day 9 of pregnancy was monitored by an aryl hydrocarbon receptor (Ah receptor, AhR)-mediated LacZ reporter system in yeast. Results showed that the number of implanted embryos was significantly reduced on day 9 of gestation by 50 and 100 ng/kg TCDD exposure. The number of implantation sites was lower for animals exposed to TCDD on days 1-3 versus those exposed during days 4-8. Decidualization in pseudopregnant mice was also inhibited by TCDD exposure. TCDD concentrations as low as 2 ng/kg significantly decreased serum progesterone levels but had no effect on serum estradiol. TCDD level in the uterus was equal to levels in the liver, but lower than the fat tissue. These results suggest that TCDD sensitivity might be attributed its local accumulation in the uterus.

Animals↗

Inhibition of HIV-1 maturation via drug association with the viral Gag protein in immature HIV-1 particles.

The small molecule 3-O-(3',3'-dimethylsuccinyl)-betulinic acid (DSB) potently inhibits human immunodeficiency virus, type 1 (HIV-1) replication by interfering with proteolytic cleavage of the viral Gag protein at a specific site. Here we have demonstrated that the antiviral mechanism involves the association of DSB with Gag at a 1:1 stoichiometry within immature HIV-1 particles. The binding was specific, as mutations in Gag that confer resistance to DSB inhibited the association, which could be competed by DSB but not by the inactive compound betulinic acid. The addition of DSB to purified immature viral cores inhibited the cleavage of Gag at the CA-SP1 junction in vitro, thus reproducing the effect of the drug when present during maturation of HIV-1 particles. Based on these findings, we propose a model in which a trimer of DSB associates with the CA-SP1 junction of adjacent subunits within the Gag polymer. The model may explain the ability of highly similar compounds to specifically target the seemingly unrelated steps of HIV-1 maturation and virus entry.

Cell Line↗

Serial measurement of serum basic fibroblast growth factor in patients with acute cerebral infarction.

Basic fibroblast growth factor (bFGF) has been reported to be involved in the pathophysiological changes following cerebral infarction. Basic fibroblast growth factor is upregulated in the brain and conduces to neuroprotection and angiogenesis in experimental brain ischemia, but the change of serum bFGF in cerebral infarction patients has not been reported. In the present study, we investigated the dynamic changes of serum bFGF in 30 patients with acute cerebral infarction and found that serum bFGF increased significantly after cerebral infarction compared with the control group (p<0.05). Serum bFGF peaked on day 3 (15.46 +/- 5.58 pg/ml; p<0.01) and remained significantly elevated on day 14 following cerebral infarction. In this study, it was also found that the levels of bFGF with large infarction were higher at each time point than those with moderate or small infarction (p<0.05). There was a positive correlation between the peak level of bFGF and improvement of clinical neurological deficits scored by Scandinavian Stroke Scale (SSS) (r=0.596; p<0.05). These results suggest that the serum bFGF level increased significantly after cerebral infarction and the level of serum bFGF could be of value to estimate the infarction size and clinical prognosis.

Acute Disease↗

Tissue distribution of a plasmid DNA containing epitopes of foot-and-mouth disease virus in mice.

It is known that only the minority of plasmid DNAs effect a cure or prevention after intramuscular injection into host. But what is the fate of the majority? And indeed how many of the injected DNAs work? Till now, little is known about it. To answer these questions, two methods including PCR and autoradiography were used in distribution study in mice that had received a single muscular inoculation of plasmid DNA containing antigenic epitopes of foot-and-mouth disease virus. The results showed that the plasmid DNAs were distributed by blood circulation and degraded soon. The degradation ratio of super coiled plasmid DNA was 20.9% in 10 min, 34.1% in 1h, 86.8% in 1 day and 97.8% in 1 week in sera in vivo. And over a half of the whole were output in urine and faeces. The rest resided most in muscles as 'antigen pool', next in immune organs, kidney, liver, heart, lung and little in brain or gonad. About 40% or 0.5% of total plasmid DNAs, inferring to be effective, resided in muscles or immune organs, respectively. Collective results suggested that 'nude' DNA, as water injection, was characterized as quick absorbent, extensive distribution, but low utilization rate. Finally, the immune mechanism for the DNA vaccine was discussed.

Animals↗

Anti-inflammatory effects of inhibiting the amine oxidase activity of semicarbazide-sensitive amine oxidase.

Human semicarbazide-sensitive amine oxidase (SSAO) or vascular adhesion protein-1 (VAP-1) is a copper-containing amine oxidase (AOC3, EC 1.4.3.6) that has both enzymatic and adhesive function. SSAO catalyzes the oxidative deamination of primary amines, resulting in the formation of the corresponding aldehyde and release of hydrogen peroxide and ammonia. Membrane-bound SSAO is an inflammation-inducible endothelial cell adhesion molecule that mediates the interaction between leukocytes and activated endothelial cells in inflamed vessels. Both the direct adhesive and enzymatic functions seem to be involved in the adhesion cascade. LJP 1207 [N'-(2-phenyl-allyl)-hydrazine hydrochloride] is a potent (human SSAO IC(50) = 17 nM), selective, and orally available SSAO inhibitor that blocks both the enzymatic and adhesion functions of SSAO/VAP-1. In a mouse model of ulcerative colitis, LJP 1207 significantly reduces mortality, loss of body weight, and colonic cytokine levels. Quantitative histopathological assessment of colitis activity in this model showed a highly significant suppression of inflammation, injury, and ulceration scores in the animals treated with the SSAO/VAP-1 inhibitor. LJP 1207 also reduced serum levels of tumor necrosis factor-alpha and interleukin 6 in lipopolysaccharide (LPS)-challenged mice and prolonged survival post-LPS-induced endotoxemia. Therapeutic and prophylactic administration of LJP 1207 in the rat carrageenan footpad model also markedly inhibited swelling and inflammation. Overall, the data suggest that small molecule SSAO/VAP-1 inhibitors may provide clinical benefit in the treatment of acute and chronic inflammatory diseases.

Amine Oxidase (Copper-Containing)↗

The preliminary efficacy of interferon-alpha and ribavirin combination treatment of chronic hepatitis C in HIV-infected patients.

BACKGROUND: It is internationally accepted that in drug-naïve individuals with hepatitis C virus (HCV) and human immunodeficiency virus (HIV) co-infection, chronic hepatitis C should be treated first if the CD4 cell count does not require the initiation of anti-retroviral therapy. Present paper evaluated the clinical effect and side-effect of interferon-alpha (IFN-alpha) and ribavirin (RBV) combination therapy for Chinese patients with HCV-HIV co-infection, and compared with them for HIV infection alone. METHODS: Ten patients with HCV-HIV and 17 patients with HCV received 5 million unit IFNalpha-2b every other day intramuscularly, and 300 mg RBV triple daily by oral. Dynamic observations were made for HCV RNA and HIV RNA loads, CD4+ and CD8+ T lymphocyte counts, liver function and blood cell measurement, and the medicine side-effects. RESULTS: After 12-week and 24-week treatments of IFN-alpha and RBV combination therapy, mean HCV RNA levels reduced 1.14 logs and 1.56 logs from the baseline at week 0 in HCV-HIV co-infection, and reduced 1.48 logs and 1.75 logs in HCV infection, respectively. The HIV RNA levels decreased 1.22 logs and 1.32 logs from the base line; however, there were no obvious different changes at T lymphocyte counts of HCV-HIV and HCV patients through 24-week treatments. Whole 27 patients showed satisfactory biochemical response to therapy. There were some mild or mediate influence-like symptoms, intestinal uncomfortable and depressed blood cell counts in early stage of the treatments. No neuropsychiatric and auto-immune disorders were found. CONCLUSIONS: IFN-alpha and RBV combination therapy had similar anti-HCV effects during 24-week treatment for HCV-HIV and HCV infected Chinese patients, and some anti-HIV effect. There were no obvious different biochemical responses and side-effects between two groups above.

Adult↗

Development of a virus-induced gene-silencing system for hexaploid wheat and its use in functional analysis of the Lr21-mediated leaf rust resistance pathway.

Virus-induced gene silencing (VIGS) is an important tool for the analysis of gene function in plants. In VIGS, viruses engineered to carry sequences derived from plant gene transcripts activate the host's sequence-specific RNA degradation system. This mechanism targets the RNAs of the viral genome for degradation, and as the virus contains transcribed plant sequence, homologous host mRNAs are also targeted for destruction. While routinely used in some dicots, no VIGS system was known for monocot plants until the recent report of silencing in barley (Hordeum vulgare) by barley stripe mosaic virus (BSMV). Here, we report development of protocols for use of BSMV to efficiently silence genes in hexaploid wheat (Triticum aestivum). The VIGS system was first optimized in studies silencing phytoene desaturase expression. Next, we used it to assay genes functioning in leaf rust resistance mediated by Lr21, which encodes a nucleotide binding site-leucine-rich repeat class resistance gene product. We demonstrated that infection with BSMV constructs carrying a 150-bp fragment of Lr21 caused conversion of incompatible interactions to compatible, whereas infection with a control construct or one that silences phytoene desaturase had no effect on resistance or susceptibility. Additionally, silencing the RAR1, SGT1, and HSP90 genes, known to be required in many but not all nucleotide binding site-leucine-rich repeat resistance pathways in diverse plant species, resulted in conversion to compatibility, indicating that these genes are essential in Lr21-mediated resistance. These studies indicate that BSMV-VIGS is a powerful tool for dissecting the genetic pathways of disease resistance in hexaploid wheat.

Gene Expression Regulation, Plant↗