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Biomedical subjects

J Hou

Publications and source records attributed to J Hou.

At least 19 recordsLinked to original sources

A polysaccharopeptide complex and a condensed tannin with antioxidant activity from dried rose (Rosa rugosa) flowers.

In this study, the fraction (P) from an aqueous extract of dried rose (Rosa rugosa) flowers was obtained by ethanol precipitation. P was chromatographed on DEAE-cellulose. The components retained on DEAE-cellulose were eluted with a linear gradient of 0-2 M NaCl solution. Two fractions, eluted at concentrations of 0.5 M NaCl and 1 M NaCl, respectively, were obtained. These two components were designated as P1 and P2, respectively. P1 was further purified using gel filtration on Sephadex G-200. P(1) yielded two peaks, and the two components were designated as P(1-a) and P(1-b), respectively. P(1-a) was a polysaccharide-peptide complex, and P(1-b) exhibited chemical properties of a condensed tannin as revealed by FTIR and NMR assay of carbohydrate and protein contents and HPLC-ESI-MS. The molecular masses of P(1-a) and P(1-b) were 150 kDa and 8 kDa, respectively. Both P(1-a) and P(1-b) possessed antioxidant activity, with the activity of P(1-b) higher than that of P(1-a). This study demonstrated that different components from rose flowers exhibited antioxidant activity.

Animals↗

Resonance contributions to anti-Stokes/Stokes ratios under surface enhanced Raman scattering conditions.

Anti-Stokes/Stokes asymmetries under surface enhanced Raman scattering (SERS) conditions are studied for a wide variety of SERS-active media and different analytes. Evidence is provided for the existence of underlying resonances that create these asymmetries. We show here that these resonances are associated with the electromagnetic coupling between the analyte (probe) and the metal. The work demonstrates the use of the anti-Stokes/Stokes ratio as a tool to understand the hierarchy of resonances in the SERS problem, which is essential for quantification purposes.

Journal Article↗

Expression and characterization of a serine protease that preferentially cleaves insulin-like growth factor binding protein-5.

Insulin-like growth factor binding proteins (IGFBPs) play important roles in regulating the functions of insulin-like growth factors (IGFs). Because IGFBPs have very high affinity for IGF-I and IGF-II, they can regulate the amount of each growth factor that is able to bind to cell surface receptors, therefore, factors that alter IGFBP affinity have the capacity to regulate IGF actions. Protease activities that are present in cell culture systems and physiologic fluids have been shown to degrade IGFBP-5. Previously, a region of sequence in a serine protease was identified that was homologous with the N-terminal 90 amino acids of members of the IGFBP family and with members of the CCN family of proteins. In a prior study, the protease was expressed in human kidney cultured cells and the cell culture supernatants were shown to cleave IGFBP-5, however, it is unknown whether the purified protease would cleave IGFBP-5 and whether it would also cleave other specific forms of IGFBPs. In this study, we expressed this protease in an insect cell expression system, purified it to homogeneity and tested its capacity to cleave IGFBP-5. The expressed protease preferentially cleaved IGFBP-5, and it had minimal activity toward other forms of IGFBPs. The proteolytic activity of this IGFBPase is inhibited by serine protease inhibitors including PMSF and 3,4-dichloroisocoumarin, as well as by divalent metal ions such as, Zn and Cu. Mutation of the active site serine resulted in a major reduction in IGFBP-5 cleavage. The protease binds to heparin and its ability to degrade IGFBP-5 is blocked in the presence of heparin. Inhibition of the activity of the protease following its secretion by B104 cells resulted in inhibition of IGFBP-5 proteolysis and IGF-I stimulation of protein synthesis. Northern blotting revealed that the transcript was expressed in multiple human tissues, including placenta, uterus, prostate, testis, spinal cord, brain, liver, small intestine, thyroid, and spleen. The highest expression was in uterus and placenta, suggesting a possible role of sex steroids in regulating its expression. Understanding the mechanism of how cleavage of IGFBP-5 by this protease alters its activity will help to further our understanding of the biologic actions of the IGFs.

Base Sequence↗

DNA methylation patterns in in vitro-fertilised goat zygotes.

Recent studies have shown that zygote demethylation patterns in mammals are variable among species. However, the methylation patterns of goat zygotes have not yet been reported on. In the present study, using immunofluorescence staining with an antibody against 5-methylcytosine, the methylation patterns of in vitro-derived goat zygotes were studied. The results indicate that goat zygotes do not undergo active global demethylation during pronuclei development, which is similar to the situation in ovine, but not in murine or bovine zygotes. This is believed to be the first report regarding methylation in goat zygotes.

Animals↗

Hot-spot mutations in hepatitis B virus core gene: eliciting or evading immune clearance?

The biological implications of substitutions L60V and I97L in the core (c) gene of hepatitis B virus (HBV) were investigated in order to determine whether they could change the immunogenicity of HBcAg or influence the immune response in mice. Three strains of recombinant adenoviruses--AdHBV-WT, AdHBV-L60V and AdHBV-I97L--containing wild-type or mutant HBV genomes were constructed using the AdEasy system and used to infect BALB/c mice intranasally. Infected mice produced anti-HBc efficiently to comparable levels. IgG1 and IgG2a specific for HBcAg were present in mice sera, and the response was dominated by IgG2a. The lymphocyte proliferative response specific for HBcAg was assessed by [3H]-thymidine uptake. We found that AdHBV-WT induced a stronger T-cell proliferation response than AdHBV-L60V and AdHBV-I97L. In conclusion, the L60V and I97L substitutions had no influence on humoral immune responses, but could downregulate T-cell responses to HBcAg, suggesting that L60V and I97L were immune escape mutants.

Animals↗

Geographic distribution, virologic and clinical characteristics of hepatitis B virus genotypes in China.

The significance of hepatitis B virus (HBV) genotypes for the heterogeneity of chronic HBV infection and severity of liver disease is not well understood. The aim of this study was to determine the distribution and virologic characteristics of HBV genotypes in China and possible association with the diversity of liver disease. The study includes 1096 chronic HBV carriers from nine provinces in China. We collected clinical and laboratory data and analysed the HBV strains in sera by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and nucleotide sequencing techniques. The most common HBV genotypes were B (41%) and C (53%), while genotypes A and D were also found. A North-South divide was identified in genotype B and C distribution - genotype C was predominant in northern China, while genotype B was more prevalent in southern provinces. Patients with genotype B were younger than those with genotype C, and had a lower prevalence of HBeAg - 65%vs 72%, respectively (P = 0.03). However, the severity of liver disease did not differ significantly between patients infected with genotype B or C - neither when comparing liver function tests (1024 patients), nor hepatic inflammation and fibrosis (264 patients). Amongst 47 patients with genotype D (by PCR-RFLP), 37 (79%) were infected with a new subtype (designated Dc), having a recombination fragment from genotype C precore/core region. This is the first large-scale HBV genotype study from China and convincing documentation of the North-to-South gradient of genotypes C vs B in this country. HBV DNA recombination over the surface and precore/core genes increases the diversity of HBV strains and may have diagnostic and clinical implications.

Adult↗

Surgical treatment for cancer of the oesophagus and gastric cardia in Hebei, China.

BACKGROUND: Southern Hebei Province has one of the highest incidences of oesophageal cancer in the world. This study describes a single-centre experience with oesophagectomy for this condition. METHODS: The reported articles on surgical treatment for cancer of the oesophagus and gastric cardia from a single department between September 1952 and December 2000 were summarized and the results compared. RESULTS: Between September 1952 and December 2000, the resectability rate increased from 78.8 to 96.0 per cent for oesophageal cancer and from 69.6 to 94.8 per cent for cancer of the cardia. The surgical mortality rate decreased from 4.6 to 1.1 per cent. There was little change in the overall 5-year survival rate; it increased from 21.2 to 23.9 per cent for patients with oesophageal cancer and from 15.2 to 17.6 per cent for those with cancer of the cardia. CONCLUSION: The rate of postoperative complications and deaths following oesophagectomy for cancer has fallen steadily over the past five decades but long-term survival remains disappointing. Improved survival is likely to be dependent on earlier diagnosis and adjunctive therapies.

Cardia↗

Transgenic twin lambs cloned by granulosa cells.

The development potential of transgenic adult cells after nuclear transfer (NT) was evaluated. Primary ovine granulosa cells (GC(S)) from a slaughter ovary were transfected with pEGFP-N1 plasmid DNA. Three G418-resistance cell lines (A2, B2 and B4) were used as donor cells in NT. A total of 162 NT blastocysts were then frozen with ethylene glycol solution and stored for five months before transplanted into recipients. Twenty-nine frozen thawed NT blastocysts were transferred into 15 synchronized recipients. Twin lambs (6.9%) derived from B2 line were delivered by cesarean section on day 143 but died after birth. A tumor consisting of lung tissues was found on the surface of left lung of the 4-kg lamb and histological analysis indicated that it resembles a hamartoma. DNA analysis confirmed that two lambs were genetically identical to B2 donor cells. Gene insertion and expression have been detected in fibroblasts cells derived from muscle tissues of the lambs. This study indicates that granulosa cell is a suitable cell type for producing transgenic animals by nuclear transfer. Offspring were produced after long-term storage of NT blastocysts.

Animals↗

A gene therapy for cancer based on the angiogenesis inhibitor, vasostatin.

The growth and persistence of solid tumors and their metastasis are angiogenesis-dependent. Vasostatin, the N-terminal domain of calreticulin inclusive of amino acids 1-180, is a potent angiogenesis inhibitor. To investigate whether intramuscular administration of vasostatin gene has the antitumor activity in mouse tumor models, we constructed a plasmid DNA encoding vasostatin and a control vector. Production and secretion of vasostatin protein by COS cells transfected with the plasmid DNA encoding vasostatin (pSecTag2B-vaso) were confirmed by Western blot analysis and ELISA. Conditioned medium from vasostatin-transfected COS cells apparently inhibited human umbilical vein endothelial cell (HUVEC) and mouse endothelial cell (SVEC4-10) proliferation, compared with conditioned medium from the COS cells transfected with control vector or non-transfected cells. Treatment with pSecTag2B-vaso twice weekly for 4 weeks resulted in the inhibition of tumor growth and the prolongation of the survival of tumor-bearing mice. The sustained high level of vasostatin protein in serum could be identified in ELISA. Angiogenesis was apparently inhibited in tumor by immunohistochemical analysis. Angiogenesis was also inhibited in the chicken embryo CAM assay and mouse corneal micropocket assay. The increased apoptotic cells were found within the tumor tissues from the mice treated with plasmid DNA encoding vasostatin. Taken together, the data in the present study indicate that the cancer gene therapy by the intramuscular delivery of plasmid DNA encoding vasostatin, is effective in the inhibition of the systemic angiogenesis and tumor growth in murine models. The present findings also provide further evidence of the anti-tumor effects of the vasostatin, and may be of importance for the further exploration of the application of this molecule in the treatment of cancer.

Angiogenesis Inhibitors↗

Thymosin-alpha1 and famciclovir combination therapy activates T-cell response in patients with chronic hepatitis B virus infection in immune-tolerant phase.

We examined whether combination therapy with thymosin-alpha1 and famciclovir would induce hepatitis B e antigen seroconversion in patients with chronic hepatitis B infection in the immune-tolerant phase without inducing significant hepatic necro-inflammation. We studied 32 hepatitis B e antigen positive patients in the immune-tolerant phase of infection, treated with 26-weeks combination therapy of famciclovir and thymosin-alpha1 (group 1). Thirty-two patients who received 26-weeks famciclovir monotherapy (group 2) and another 32 patients who received no treatment (group 3), served as controls. Peripheral blood mononuclear cell proliferation and cytokine secretion in response to recombinant HBV core and surface antigen and serial serum HBV-DNA, were assayed. No significant difference in adverse events were observed among the three groups. By week 26, the median reduction in group 1 (0.94 log10 copies/mL) was greater than group 2 (0.70 log10 copies/mL, P < 0.001). Five (15.6%) patients in group 1 at 52 weeks (median range 13-78 weeks) and none in group 2 or 3 experienced hepatitis B e antigen seroconversion (P = 0.053). Sustained serological clearance of hepatitis B e antigen was associated with activation of CD4 positive HBV-specific T-cell reactivity and were of T-helper 1. Hence combination therapy with immunomodulatory agents and nucleoside analogues should be explored.

2-Aminopurine↗

Neural correlates of response inhibition for behavioral regulation in humans assessed by functional magnetic resonance imaging.

To identify the specific frontal and cingulate regions involved in response inhibition, five Chinese right-handed male volunteers with an attention span reaching or exceeding a 7-digit level participated in this blocked design functional magnetic resonance imaging study. Each block represented one of the two experimental conditions, the Correct Matching or the Incorrect Matching condition. In the Correct Matching condition, the subjects were required to make correct judgement of whether the two 3-digit numbers presented in succession were the same or not; whereas in the Incorrect Matching condition, the participants were instructed to inhibit the correct responses. Bilateral activations of the prefrontal and anterior cingulate as well as the left posterior cingulated cortex were observed while the subjects were exercising response inhibition. The roles of these activations in response inhibition were discussed.

Adult↗

Perfusion-weighted imaging of interictal hypoperfusion in temporal lobe epilepsy using FAIR-HASTE: comparison with H(2)(15)O PET measurements.

To detect perfusion abnormalities in areas of high magnetic susceptibility in the brain, an arterial spin-labeling MRI technique utilizing flow-sensitive alternating inversion recovery (FAIR) and half-Fourier single shot turbo spin-echo (HASTE) for spin preparation and image acquisition, respectively, was developed. It was initially tested in a functional study involving visual stimulation, and was able to detect significant activation with an increase (approximately 70%) in relative cerebral blood flow. Subsequently, it was applied in a clinical situation in eight patients with temporal lobe epilepsy (TLE). The perfusion-weighted images obtained showed no susceptibility artifacts even in the region of the inferior temporal lobe and were able to detect interictal hypoperfusion in TLE. The results were compared with those derived from H(2)(15)O PET perfusion imaging in each patient. A statistically significant correlation (r = 0.75, P < 0.05) was found between results acquired from these two modalities. Magn Reson Med 45:431-435, 2001.

Artifacts↗

Effect of smoking on serum concentrations of total homocysteine and B vitamins in mid-pregnancy.

There are conflicting findings in the literature on the effect of smoking on total homocysteine (tHcy) concentrations in non-pregnant subjects. We evaluated the effect of smoking on serum concentrations of tHcy, folate, vitamin B-12 pyridoxal 5'-phosphate (PLP, a coenzyme form of vitamin B-6) in 196 women at 18 and 30 weeks' gestation. The smokers were defined as those who self-reported cigarette smoking and had serum concentrations of thiocyanate, a biomaker of smoking, in the highest quartiles of the population. Mid-pregnancy serum tHcy concentrations were not significantly different between smokers and non-smokers. Folate, vitamin B-12 and PLP concentrations were generally lower in smokers than non-smokers. In smokers, tHcy concentrations had significant negative correlations with folate at both time points. The multiple regression analyses indicated that serum folate concentration was the most significant factor associated with tHcy concentrations among smokers, whereas thiocyanate concentrations showed no such effect. We conclude that serum tHcy concentrations were most strongly associated with the nutritional status of folate among the B vitamins tested during mid-pregnancy in our subjects. We suggest that it is essential to consider the nutritional status of folate, vitamin B-12 and vitamin B-6 in evaluating the effect of smoking on homocysteine metabolism.

Female↗

Gene therapy for hepatocellular carcinoma based on tumour-selective suicide gene expression using the alpha-fetoprotein (AFP) enhancer and a housekeeping gene promoter.

The aim of this study was to examine whether the human alpha-fetoprotein (AFP) enhancer could be used to induce hepatocellular carcinoma (HCC)-selective expression of the herpes simplex virus thymidine kinase (HSV-tk) gene which is under the control of the phosphoglycerate kinase (pgk) promotor. The human AFP enhancer was linked with the non-tissue-specific, human housekeeping pgk promoter in a retroviral vector. AFP-producing HCC cells infected with retroviruses carrying the HSV-tk gene under the control of the AFP enhancer/pgk promoter were much more susceptible to the prodrug, ganciclovir (GCV), than those infected with the same retroviruses without the AFP enhancer. Non-HCC cells infected with retroviruses carrying the HSV-tk gene under the control of the AFP enhancer/pgk promoter exhibited profoundly increased resistance to GCV compared with those infected with the same retroviruses without the AFP enhancer. Northern blot analysis revealed that the AFP enhancer caused enhanced HSV-tk expression in AFP-producing HCC cells and suppressed HSV-tk expression in non-HCC cells. Our results indicate that the AFP enhancer could give HCC selectivity to the pgk promoter, and that this novel strategy may be useful for HCC-selective cancer gene therapy.

Blotting, Northern↗

Cold adaptive thermogenesis in small mammals from different geographical zones of China.

The mechanisms of thermogenesis and thermoregulation were studied in the tree shrew (Tupaia belangeri) and greater vole (Eothenomys miletus) of the subtropical region, and Brandt's vole (Microtus brandti), Mongolian gerbil (Meriones unguiculatus), Daurian ground squirrel (Spermophilus dauricus) and plateau pika (Ochotona curzoniae) of the northern temperate zone. Resting metabolic rate (RMR) and non-shivering thermogenesis (NST) increased significantly in T. belangeri, E. miletus, M. brandti and M. unguiculatus after cold acclimation (4 degrees C) for 4 weeks. In T. belangeri, the increase in RMR and thermogenesis at liver cellular level were responsible for enhancing the capacity of enduring cold stress, and homeothermia was simultaneously extended. Stable body temperature in M. brandti, E. miletus, M. unguiculatus and O. curzoniae was maintained mainly through increase in NST, brown adipose tissue (BAT) mass and its mitochondrial protein content, and the upregulation of uncoupling protein (UCP1) mRNA, as well as enhancement of the activity of cytochrome C oxidase, alpha-glycerophosphate oxidase and T(4) 5'-deiodinase in BAT mitochondria. The RMR in O. curzoniae and euthermic S. dauricus was not changed, while NST significantly increased during cold exposure; the former maintained their stable body temperature and mass, while body temperature in the latter declined by 4.8 degrees C. The serum T(3) concentration or ratio of T(3)/T(4) in all the species was enhanced after cold acclimation. Results indicated that: (1) the adaptive mechanisms of T. belangeri residing in the subtropical region to cold are primarily by increasing RMR and secondly by increasing NST, and the mechanisms of thermogenesis are similar to those in tropical mammals; (2) in small mammals residing in northern regions, the adaptation to cold is chiefly to increase NST; (3) the mechanism of cold-induced thermogenesis in E. miletus residing in subtropical and high mountain regions is similar to that in the north; (4) a low RMR in warm environments and peak RMR and NST in cold environments enabled M. unguiculatus to tolerate a semi-desert climate; (5) O. curzoniae has unusually high RMR and high NST, acting mainly via increasing NST to adapt to extreme cold of the Qinghai-Tibet Plateau; (6) the adaptation of euthermic S. dauricus to cold is due to an increase in NST and a relaxed homeothermia; and lastly (7) the thyroid hormone is involved in the regulation of cold adaptive thermogenesis in all the species studied.

Adaptation, Physiological↗