Biomedical subjects

P Hu

Publications and source records attributed to P Hu.

At least 109 records · Page 6Linked to original sources

PubMed1999

[Long-term effect of extra-trabeculectomy in treatment of congenital glaucoma].

OBJECTIVE: To assess the long-term effect of extra-trabeculectomy in treatment of congenital glaucoma. METHODS: 29 patients (47 eyes) with congenital glaucoma having undergone extra-trabeculectomy were followed up for over 5 years. RESULTS: Success rates after single surgery were 70.2% at 6 months, 63.8% at 12 months, 57.5% at 18 months and 55.3% at 24 months. Re-elevation of intraocular pressure (IOP) all happened within 2 years after surgery. 14 eyes underwent second trabeculectomy, and 7 eyes (50.0%) were successful the total success rate was 70.2%. The naked visual acuity of 12 eyes (36.0%) among successful eyes was >or= 0.5. All the diseased eyes were myopic (-0.50 to -25.00 D). CONCLUSION: Follow-up of trabeculectomy should be emphasized within 2 years after surgery. Ametropic and anisometropic amblyopia should be corrected promptly after surgery.

Child, Preschool↗

PubMed1999

[The effect of simulated weightlessness on the function of spleen lymphocytes and the expression of c-fos proto-oncogene in tail-suspended mice].

Objective. To investigate the effect of simulated weightlessness on proliferation of spleen lymphocyte, production of interleukin-2 and expression of c-fos proto-oncogene in mice. Method. Mice were tail-suspended for 7 or 14 days to simulate the effect of weightlessness; proliferation of lymphocyte was measured by MTT method, production of IL-2 was determined by biological activity and expression of c-fos proto-oncogene was determined by dot blotting. Result. As compared with the control, the proliferation of spleen lymphocyte and the production of IL-2 were significantly decreased in 14 d suspended group, but the expression of c-fos proto-oncogene [correction of oncogen] was increased in both 7 d and 14 d suspended groups. Conclusion. This result suggested that simulated weightlessness could interfere with the genes regulating the lymphocyte.

Animals↗

PubMed1999

[Relationship between expression of estrogen receptor progestrone receptor and the biological characteristics of nasopharyngeal carcinoma].

OBJECTIVE: To Clarity the relationship between the expression of the estrogen receptor (ER), progestogen receptor (PR) and the biological characteristics of nasopharyngeal carcinoma (NPC). METHOD: The immuno-histochemical LSAB method was used to analyse the ER and PR in paraffin section specimen of 71 cases of nasopharyngeal carcinoma. CT scaning and clinic presentation were also put into consideration. RESULT: The expression of ER with (-), (+), (++) and (+++) in 71 cases were: 4(5.63%), 19(26.76%), 27(38.03%) and 21 (26.76%), respectively. The expression of PR with (-), (+) and (++) in 71 cases were: 41(57.75%), 29 (39.44%) and 2(2.82%), respectively. The encroaching of tumor to the tissue of parapharynx, the metastasis rate of lymphonode and clinic stage were related to the expression of ER (P < 0.01). The encroaching of tumor to the tissue of parapharynx, the metastasis rate of lymphonode were also assciated with the expression of PR (P < 0.05). CONCLUSION: NPC is a tumor of hightly expression of ER and PR. The positive intensity of ER and PR were associated with the encroaching extent of NPC, and the metastasis rate of lymphonode. It is significant to detarmine ER and PR of NPC in assessing the biological characterisitics of NPC and predicting the prognosis of NPC. It also suggests that the endocrine therapy can be put into consideration.

Adult↗

PubMed1999

[Prokaryotic expressed trichosanthin and other two proteins have anti-fungal activity in vitro].

DNAs encoding Trichosanthin, tobacco class I Chitinase and tobacco class I beta-1, 3-Glucanase were expressed in E. coli, respectively. The expression products were assayed for their anti-fungal activity. All of three kinds of proteins show anti-fungal activity. When two of them combined, this activity was enhanced greatly. When three of them combined, the stronger anti-fungal activity was observed.

Antifungal Agents↗

PubMed1999

[Plasma branched-chain ketoacids metabolism in patients with chronic renal failure after oral administration of ketosteril].

OBJECTIVE: To investigate the metabolism of branched-chain ketoacids (BCKA) in patients with chronic renal failure (CRF) and healthy control after an oral dose of ketosteril. METHODS: To determine plasma concentration of BCKA by gas chromatography/mass spectrography(GC/MS). RESULTS: Fasting plasma concentration of ketoisocaproate (KICA) and ketomethylvalerate(KMVA) in patients with CRF were lower than that of healthy control; The Cmax of KICA, KMVA and ketoisovalerate (KIVA) were (67.7 +/- 12.9), (57.8 +/- 10.3), (30.3 +/- 5.3) mumol/L; Tmax were (43.8 +/- 5.7), (58.1 +/- 4.6), (43.8 +/- 4.5) min; T1/2 were (108.5 +/- 12), (105 +/- 16.6), (116.8 +/- 18) min respectively. CONCLUSIONS: Absorption of BCKA in ketosteril had no significant difference between CRF patients and healthy controls; ketosteril could be used to treat CRF patients effectively.

Administration, Oral↗

PubMed1999

Pretreatment with a monoclonal antibody/interleukin-2 fusion protein directed against DNA enhances the delivery of therapeutic molecules to solid tumors.

The efficacy of molecular therapies for human malignancies is limited by inadequate accumulation within solid tumors. Our laboratory has developed a novel approach that uses monoclonal antibodies (MAbs) to direct vasoactive proteins to tumor sites to increase local vascular permeability and, in turn, improve the delivery of therapeutic reagents. Previously, we demonstrated that pretreatment with immunoconjugates containing interleukin-2 (IL-2) enhances specific tumor uptake of radiolabeled MAbs without affecting normal tissues. In the present study, we describe a fusion protein consisting of a chimeric antinuclear antibody and IL-2 (chTNT-3/IL-2) and illustrate its potential for improving the delivery of both MAbs and drugs. The ability of pretreatment with chTNT-3/IL-2 to increase specific tumor uptake of the MAb B72.3 was demonstrated in LS174T colon tumor-bearing mice. Tumor accretion of B72.3 increased nearly 3-fold, with no changes in normal tissues. Abrogation of this effect with N(G)-methyl-1-arginine, a chemical inhibitor of nitric oxide synthase, suggests that rapid generation of nitric oxide in the tumor is responsible for the enhanced uptake. To demonstrate that pretreatment with chTNT-3/IL-2 can improve the uptake of other clinically relevant MAbs in different tumor models, additional studies were performed in both lung and prostate xenograft models. Pretreatment with the fusion protein increased specific tumor uptake of the MAb NR-LU-10 in A427 lung tumor-bearing mice and enhanced tumor uptake of the MAb CYT-351 in LNCaP prostate tumor-bearing mice, 2.1-fold and 1.7-fold, respectively. Finally, tumor uptake of the radiolabeled thymidine analogue 125IUdR also increased approximately 3-fold after pretreatment, indicating that this approach can be extended to small molecules such as chemotherapeutic drugs. Because TNT-3 recognizes a universal nuclear antigen accessible in degenerating and necrotic cells within all solid tumors, this strategy may be applicable to the majority of human cancers.

Animals↗

PubMed1998

Evaluating antibodies for their capacity to induce cell-mediated lysis of malignant B cells.

Promising results from clinical trials have led to renewed interest in effector mechanisms operating in antibody-based therapy of leukemia and lymphoma. We tested a panel of B-cell antibodies from the Sixth Human Leukocyte Differentiation Antigen workshop for their capacity to mediate antibody-dependent cellular cytotoxicity, often considered to be one of the most potent effector mechanisms in vivo. As effector cells, mononuclear cells and polymorphonuclear (PMN) cells from healthy donors were compared with Fc gammaRI (CD64)-expressing PMN cells from patients receiving granulocyte colony-stimulating factor (G-CSF) treatment. Of the 29 IgG workshop antibodies binding most strongly to the tested malignant human B-cell lines, only 3 consistently induced target cell lysis. These three antibodies were determined to be HLA DR reactive. Experiments with a panel of HLA class II antibodies showed the involvement of individual Fc gamma receptors on effector cells to be strongly dependent on the antibody isotype. We then compared killing mediated by chimeric IgG1 antibodies with that from Fc gammaRI-directed bispecific antibodies, targeting classical HLA class II, or the Lym-1 and Lym-2 antigens. The latter two are variant forms of HLA class II, which are highly expressed on the surface of malignant B cells but which are found only at low levels in normal cells. With blood from G-CSF-treated donors, bispecific antibodies showed enhanced killing compared to their chimeric IgG1 derivatives, because they were more effective in recruiting Fc gammaRI-expressing PMN cells. G-CSF- and Fc gammaRI-directed bispecific antibodies to HLA class II, therefore, seem to be an attractive combination for lymphoma therapy.

Antibodies, Bispecific↗

PubMed1998

[Chromosome assignment of a novel microsatellite polymorphic marker (D14S1435) by fluorescence in situ hybridization].

OBJECTIVE: Chromosome assignment of a novel microsatellite marker (D14S1435) containing CA repeats and showing 11 alleles and 0.85 of PIC value in Chinese population) isolated from microdissected human chromosome 14q24.3 DNA library by fluorescence in situ hybridization(FISH). METHODS: The polymorphic marker was used as probe to screen a human Lambda/DASH genome library, a positive clone was isolated and the recombinant phage DNA was extracted as primary material. The insert was recovered through BamH I digestion to complete and low melting point agarose gel electro- phoresis, after processed with Sau3A I the probe was prepared by linker- catch PCR method and the following fluorescence in situ hybridization(FISH) was conducted to metaphase chromosomes. RESULTS: this novel STR was precisely mapped to human chromosome 14q24.3, and this demonstrated the validity of isolating chromosome band-specific genetic markers from probe pool made by chromosome microdissection. CONCLUSION: Confirmed by chromosome fluorescence in situ hybridization, the novel STR obtained by chromosome microdissection, PCR and subcloning method was really from the micro- dissected region. This not only increases the numbers of genetic markers used for linkage analysis in the region,but also provides a novel valuable genetic marker for the gene diagnosis and gene cloning of the possible disease-causing genes located in the region of 14q24.3.

Chromosome Mapping↗

PubMed1998

[Rapid selection of the gradually shortened DNA clones by using PCR amplification].

OBJECTIVE: To establish a simple technique with which the gradually shortened DNA clones can be selected rapidly and accurately so that the sequencing and identification of the full length cDNA or longer DNA fragments can be performed as quick as possible. METHODS: Primers were designed according to the sequence flanking the multiple cloning site of pGEM vector and were used to amplify the inserted fragments in a series of deletion sub-clones originated from a clone. with longer DNA insert fragment. These sub-clones which contain the gradually shortened fragments can be selected directly. This method was compared with the routine method in which the restriction endonuclease was used to digest DNA samples. RESULTS: A series of deletion sub-clones were identified accurately with this new method and the cycle sequencing could be performed on these PCR products as well. This method was characteristic of more accurate, more simple and less time-consuming, compared with the routine method. CONCLUSION: The method presented is very effective for rapid selection of the gradually shortened inserted fragments constructed in pGEM vector. It can greatly save time and materials, and the strategy in the report could also fit for selecting the clones containing gradually shortened fragments in other plasmid vectors.

DNA, Complementary↗

PubMed1998

Transcription rates of SERCA and phospholamban genes change in response to chronic stimulation of skeletal muscle.

Chronic low frequency stimulation of predominantly fast-twitch skeletal muscles decrease the levels of SERCA1 (fast-twitch sarco(endo)plasmic reticulum Ca2+-ATPase) mRNA, and increase the levels of SERCA2 (slow-twitch sarco(endo)plasmic reticulum Ca2+-ATPase) and phospholamban mRNAs. To assess the role of transcription in these changes in mRNA levels, nuclei were isolated from chronically stimulated canine latissimus dorsi muscles and transcription rates were estimated by nuclear run-on assays. Decreases in the rates of SERCA1 gene transcription matched the fall in its mRNA level and increases in the rates of SERCA2 and phospholamban gene transcription matched the increases in their mRNAs.

Animals↗

PubMed1998

The beta2-agonist salbutamol affects the expression of phospholamban and both isoforms of SERCA in canine skeletal muscle and blocks changes in these induced by neuromuscular stimulation.

Chronic administration of salbutamol induced expression of hybrid fibers in canine skeletal muscles. Fast-twitch fibers expressed SERCA2a (the slow-twitch isoform of sarcoplasmic reticulum Ca2+-ATPase) and slow-twitch fibers expressed SERCA1 (the fast-twitch isoform of the Ca2+-ATPase). The proportion of fibers that became hybrid increased from a small percentage in the control muscles to 30% in the predominantly fast-twitch latissimus dorsi and to 45% in the predominantly slow-twitch vastus intermedius. In contrast to this response by the SERCA genes the phospholamban gene response was muscle specific. The fraction of fibers that expressed phospholamban decreased slightly in the latissimus dorsi while increasing moderately in the vastus intermedius. The effects of chronic neurostimulation of the latissimus dorsi on SERCA1, SERCA2a and phospholamban levels were mostly blocked by salbutamol. While 100% of fibers from neurostimulated muscles expressed phospholamban, only 51% of the fibers from the neurostimulated and salbutamol-treated muscles expressed it. In the neurostimulated muscle, very few muscle fibers expressed SERCA1a while 61% of the fibers that received salbutamol expressed it, albeit as hybrid fibers. The levels of SERCA2a in response to these interventions were just the opposite. In the neurostimulated muscle 37.5% of fibers were hybrid and 62.5% expressed SERCA2a only. With co-administration of neurostimulation and salbutamol, 61.3% of fibers were hybrid and 38.7% expressed SERCA2a only.

Adrenergic alpha-2 Receptor Agonists↗

PubMed1998

A low rate of reinfection following effective therapy against Helicobacter pylori in a developing nation (China).

BACKGROUND & AIMS: In developed countries, reinfection after successful eradication of Helicobacter pylori appears unusual. High prevalences of H. pylori in developing countries may result in high reinfection rates. The aim of this study was to determine the rate of reinfection and ulcer recurrence in Chinese patients cured of H. pylori and duodenal ulcer disease. METHODS: One hundred eighty-four patients with duodenal ulcer disease shown by endoscopic examination (1 month) and 14C-urea breath test (3 months) after termination of treatment to have cleared their H. pylori were investigated. Patients were followed up by endoscopy (12 and 24 months) and breath test (6, 9, 12, 18, and 24 months). H. pylori status at endoscopic examination was determined by rapid urease, histology, and culture. In reinfected patients, random amplification of polymorphic DNA fingerprinting was used to compare isolates before and after therapy. RESULTS: Four patients were reinfected with H. pylori over 24 months (3 within 6 months and 1 at 24 months; average annual recurrence rate, 1.08%). Fingerprinting of isolates from 3 patients showed 1 patient (6 months) to have identical strains and the remainder to have nonidentical strains before and after treatment. Ulcer relapse occurred in 6 patients (4 H. pylori positive). CONCLUSIONS: Reinfection with H. pylori is rare in developing countries where treatment is effective.

Adult↗

PubMed1998

Axonal sprouts containing calcitonin gene-related peptide and substance P form pericellular baskets around large diameter neurons after sciatic nerve transection in the rat.

Trauma to a peripheral nerve is followed by anatomical remodelling proximal to the lesion, including sprouting of perivascular axons to form baskets of noradrenergic terminals ("rings") around large diameter primary afferent somata in the dorsal root ganglia containing lesioned neurons and sprouting of Abeta axons terminating deep in the dorsal horn into Lamina II. These abnormal structural associations may contribute to the changes in sensory processing that lead to neuropathic pain. The trigger for sprouting of sympathetic axons is probably a neurotrophin (such as nerve growth factor) concentrated locally. Nerve growth factor messenger RNA is known to be increased in dorsal root ganglia after sciatic nerve transection and increased local nerve growth factor elicits collateral sprouting of intact sympathetic terminals into adjacent denervated skin. Nociceptive terminals containing calcitonin gene-related peptide, with or without substance P, also sprout in response to raised concentrations of nerve growth factor. We have found that axons of small sensory neurons immunoreactive for these peptides also form rings around large diameter dorsal root ganglion cells of adult rats after sciatic nerve transection, concurrently with the development of rings of noradrenergic terminals.

Animals↗

PubMed1998

Recombinant breast carcinoma-associated mucins expressed in a baculovirus system contain a tumor specific epitope.

Mucins are highly immunogenic glycoproteins that are abundantly expressed by breast and other adenocarcinomas. In order to progress in the understanding of the structure immunity relationship of the breast tumor associated mucin and normal tissue mucin, two forms of breast carcinoma associated mucin, muc7-BV and pem-BV, were expressed in a baculovirus expression system. The muc7-BV was constructed by inserting the seven tandem repeats of mucin core cDNA fragment into transfer vector pAc360, forming a fusion protein containing 14 amino acids of the baculovirus polyhedrin N-terminus. The pem-BV was constructed by cloning full-length mucin cDNA into the transfer vector pVL1392. The recombinant mucins were purified using immunoaffinity chromatography. The purified muc7-BV and pem-BV had molecular weights of 28 and 59 kd, respectively. No carbohydrate was detected on these recombinant mucins and is speculated to explain why both forms of recombinant mucin showed strong affinity to tumor-specific monoclonal antibody SM3. These recombinant mucins may have the potential value to develop vaccines against breast and other adenocarcinomas and to induce cytotoxic T-lymphocyte lines for immunotherapy of the same.

Adenocarcinoma↗

PubMed1998

Characterization of the regional intestinal kinetics of drug efflux in rat and human intestine and in Caco-2 cells.

PURPOSE: The aim of the present study was to investigate the transport kinetics of intestinal secretory processes in the jejunum, ileum and colon of rats and humans and in Caco-2 cells, in vitro. METHODS: Etoposide, vinblastine sulphate and verapamil hydrochloride were chosen as model substrates since they have been reported to undergo efflux in various other tissues. The concentration dependence, inhibition, directionality, temperature dependence, proton/sodium dependence, and ATP dependence of efflux were studied using side-by-side diffusion chambers and brush border membrane vesicles (BBMVs). Intestinal tissue from rats and humans and Caco-2 cells (passage no. 26) were used. Directional steady state effective permeabilities were calculated from drug appearance in the apical (AP) or basolateral (BL) chambers. Kinetic studies were carried out by investigating substrate efflux at concentrations ranging from 0.2 microns to 1000 microns. Since substrate efflux may be a result of more than one transporter, the hybrid efflux Km (Michaelis-constant), Pc (carrier-mediated permeability), and Pm (passive permeability) were determined as a function of intestinal region. Inhibitor studies were performed using quinidine (0.2mM), a mixed inhibitor of P-glycoprotein (Pgp) and Multidrug Resistance-Associated Protein (MRP), and Leukotriene C4 (100 nM), an inhibitor of MRP and the canalicular multispecific organic anion transporter (cMOAT). Temperature dependent efflux was determined by investigating the BL to AP transport at temperatures ranging from 3 degrees C to 37 degrees C. Energies of activation (Ea) were determined from an Arrhenius analysis. Sodium, proton, and ATP dependence were determined using BBMVs. Immunoquantitation of Pgp, MRP and Lung Resistance Protein (LRP) in Caco-2 cells were carried out using Western blot analysis. RESULTS: Active efflux of all substrates was observed in all regions of rat and human intestine and in Caco-2 cells. Directionality was observed with BL to AP transport exceeding AP to BL transport. The BL to AP/AP to BL permeability ratio, the efflux ratio, ranged from 1.4 to 19.8. Ileal efflux was significantly higher (p < 0.001) than in other regions. Kinetic studies revealed that hybrid efflux Km values ranged from 4 to 350 microns. In some cases, efflux was not saturable due to the solubility limits of the compounds utilized in this study. In presence of inhibitors, efflux ratios approached 1. BL to AP transport was temperature dependent in rat ileum for all substrates. Each of the intestinal efflux was found to be 11.6, 8.3, and 15.8 kcal/mole for etoposide, vinblastine and verapamil, respectively, suggesting an active, energy-dependent efflux mechanism. Substrate efflux was not sodium or proton dependent but was dependent on ATP. Using Western blot analysis the presence of Pgp, MRP, and LRP was demonstrated in Caco-2 cells and the amount of each transport protein varied as a function of passage number. CONCLUSIONS: Using multiple putative efflux substrates, the current results demonstrate that intestinal efflux was regionally dependent, mediated by multiple efflux transporters, the Km's were in the micro-molar range, and involved an energy dependent mechanism(s).

Adenosine Triphosphate↗

PubMed1998

Effect of baffles and a louvered bypass on the airflow and the convective patterns of contaminant inside a fume hood.

Various numerical predictions are given for the containment testing of fume hoods with internal baffles of different size and at different locations and with a louvered bypass. The numerical predictions were compared with experimental data and a fairly good agreement for the air velocity was found, although there was only qualitative agreement for the concentration of the contaminant. It was found that the level of contaminant leakage at the working aperture depended not only on the size of the recirculating airflow behind the fume hood sash but also on the blockage caused by the baffles inside the fume hood near the exhaust duct. Finally, some new suggestions for the design of the fume hood have been proposed. However, before implementing these suggestions it is essential that either experimental testing be performed, or the cost-effective alternative of using numerical predictions for containment testing, to determine accurately the level of contaminant leakage from the working aperture.

Air Movements↗

PubMed1998

A new chemically modified chimeric TNT-3 monoclonal antibody directed against DNA for the radioimmunotherapy of solid tumors.

In the last several years, our laboratory has developed a new approach to the radioimmunotherapy of solid tumors, designated Tumor Necrosis Treatment (TNT), that exploits the presence of degenerating and necrotic cells within tumors by utilizing MAbs directed against universal, intracellular antigens. The first TNT MAb developed by our laboratory, designated TNT-1, was directed against nucleosomal determinants consisting of histone H1 and DNA. Since absolute tumor accretion of MAb is a critical determinant of antitumor efficacy in radioimmunotherapy, we sought to identify new antinuclear antibodies that displayed high tumor localization properties. In the present study, we describe a murine antinuclear antibody, TNT-3, which demonstrates 3-fold higher tumor uptake than TNT-1. Because of this characteristic, a chimeric derivative designated chTNT-3 was developed and evaluated for antigen binding and tumor targeting. ELISA studies using a series of nuclear antigens confirmed that TNT-3 is directed against single-stranded DNA and does not cross react with TNT-1. Immunohistology reveals predominantly nuclear staining reactivity in human tissues and tumors. Since it was shown by our laboratory that charge modification can significantly improve the pharmacokinetic performance of monoclonal antibodies, chTNT-3 was chemically modified with biotin to generate an improved therapeutic reagent designated chTNT-3/B. Comparative studies with unmodified MAb demonstrated that biotinylation significantly shortened its clearance time in mice and produced lower normal tissue levels, while maintaining an equal amount of uptake in tumor xenografts for up to 10 days. These in vivo characteristics suggest that chTNT-3/B is an improved TNT reagent for the radioimmunotherapy of solid tumors.

Animals↗

PubMed1998

Microvascular and cellular responses in the optic nerve of rats with acute experimental allergic encephalomyelitis (EAE).

The optic nerve of rats with EAE was examined at various times to determine the integrity of the blood-brain barrier (BBB) and to assess monocyte-macrophage, T cell, and microglial responses. In naive control animals, leakage of horseradish peroxidase (HRP) and the presence of cells expressing major histocompatibility complex (MHC) class II antigen were evident in the meninges of the retrobulbar optic nerve. In rats with EAE, microglia in the region of the lamina cribrosa and in the regions adjacent to the meninges became activated from day 7 to 8 postinduction (pi). HRP leakage was also evident in the region of the lamina cribrosa from day 7 to 8 pi. On day 8 pi, infiltration of inflammatory cells and Monastral blue leakage were apparent in the myelinated region of the optic nerve. The intensity of these cellular and vascular changes peaked at day 12 pi, when signs of clinical disease became manifest. Monocytes-macrophages expressing MHC class II and the ED1 antigen, together with lymphocytes expressing the alphabetaT cell receptor, constituted the major proportion of cells associated with inflammatory lesions. Thus: (i) the inherent weakness of the BBB as well as the presence of both antigen (myelin) and MHC class II+ cells in the retrobulbar optic nerve are likely susceptibility factors for the frequent involvement of this region in EAE and multiple sclerosis; and (ii) activation of microglia occurs early in the pathogenesis of experimental optic neuritis.

Animals↗