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H Fang

Publications and source records attributed to H Fang.

At least 55 records · Page 3Linked to original sources

[The key enzymes of metabolisms of glycerol in Candida glycerolgenesis].

The specific enzymes for glycerol formation and dissimilation in Candida glycerolgenesis were studied. C. glycerolgenesis hardly assimilated glycerol without glucose and a little amount of glucose remarkably stimulated this dissimilation pathway and the activity and expression of mitochondrial glycerol-3-phosphate dehydrogenase was induced strongly by glycerol-3-phosphate and repressed by glucose metabolism. During glycerol production fermented by C. glycerolgenesis, the activity of cytoplasmic glycerol 3-phosphate dehydrogenase maintained at a higher level and existed two peaks of activity, which appeared at 36 h and 60 h, respectively. The activity level of the first activity peak determines glycerol productivity and becomes a key enzyme of glycerol formation. The activity of glycerol 3-phosphatase maintained at a higher level just between 18 h and 48 h and appeared on peak activity at 36 h. In that period, the activity of glycerol 3-phosphatase was up to more than two times of cytoplasmic glycerol 3-phosphate dehydrogenase and that was inosculated with the phase of rapid accumulation of glycerol during glycerol production by C. glycerolgenesis. In the phase of slowly accumulation of glycerol between 48 h and 72 h, the activity of glycerol 3-phosphatase decreased below that of cytoplasmic glycerol 3-phosphate dehydrogenase. So, cytoplasmic glycerol 3-phosphate dehydrogenase is a key enzyme of glycerol formation in the phase of rapid accumulation of glycerol and glycerol 3-phosphatase becomes a more effective factor after glycerol fermentation reached into slowly accumulation phase. In a conclusion, it is a fundamental for C. glycerolgenesis over-producing high level of glycerol that its cytoplasmic glycerol 3-phosphate dehydrogenase maintains at high activity and the activity of glycerol 3-phosphatase is much higher than that of cytoplasmic glycerol 3-phosphate dehydrogenase.

Candida↗

Signal peptides having standard and nonstandard cleavage sites can be processed by Imp1p of the mitochondrial inner membrane protease.

We have performed a site-directed mutagenesis study showing that residues comprising the type I signal peptidase signature in the two catalytic subunits of the yeast inner membrane protease, Imp1p and Imp2p, are functionally important, consistent with the idea that these subunits contain a serine/lysine catalytic dyad. Previous studies have shown that Imp1p cleaves signal peptides having asparagine at the -1 position, which deviates from the typical signal peptide possessing a small uncharged amino acid at this position. To determine whether asparagine is responsible for the nonoverlapping substrate specificities exhibited by the inner membrane protease subunits, we have substituted asparagine with 19 amino acids in the Imp1p substrate i-cytochrome (cyt) b(2). The resulting signal peptides containing alanine, serine, cysteine, leucine, and methionine can be cleaved efficiently by Imp1p. The remaining mutant signal peptides are cleaved inefficiently or not at all. Surprisingly, none of the amino acid changes results in the recognition of i-cyt b(2) by Imp2p, whose natural substrate, i-cyt c(1), has alanine at the -1 position. The data demonstrate that (i) although the -1 residue is important in substrates recognized by Imp1p, signal peptides having standard and nonstandard cleavage sites can be processed by Imp1p, and (ii) a -1 asparagine does not govern the substrate specificity of the inner membrane protease subunits.

Amino Acid Sequence↗

Spontaneous activation of human immunodeficiency virus type 1 in an immunotoxin-resistant variant T cell line.

We have previously used immunotoxins to select for HIV-1 variant cells that transcribe HIV at extremely low levels and fail to produce HIV proteins. Further observation of one of these variants (E9) demonstrated spontaneous in vitro activation of HIV production. The mechanism of activation is different from that of U1 and ACH-2, two other cell lines that can be activated to express HIV in vitro.

Cell Line↗

The catalytic mechanism of endoplasmic reticulum signal peptidase appears to be distinct from most eubacterial signal peptidases.

Many type I signal peptidases from eubacterial cells appear to contain a serine/lysine catalytic dyad. In contrast, our data show that the signal peptidase complex from the endoplasmic reticulum lacks an apparent catalytic lysine. Instead, a serine, histidine, and two aspartic acids are important for signal peptidase activity by the Sec11p subunit of the yeast signal peptidase complex. Amino acids critical to the eubacterial signal peptidases and Sec11p are, however, positioned similarly along their primary sequences, suggesting the presence of a common structural element(s) near the catalytic sites of these enzymes.

Amino Acid Sequence↗

B7-2 is required for the progression but not the initiation of the type 2 immune response to a gastrointestinal nematode parasite.

T cells require CD28/CTLA-4 costimulatory molecule interactions in addition to Ag-specific signals through the TCR for in vivo effector Th cell function. Some studies have suggested that the ligands for these costimulatory molecules may differentially influence effector T cell function with B7-2 favoring a type 2 response and B7-1 favoring a type 1 response, while other studies have suggested that these molecules may be redundant. The recent development of B7-2-deficient mice permits the direct analysis of the requirement of B7-2 during a type 2 immune response to an infectious pathogen. We have examined, in B7-2-deficient mice, effector Th cell function and the associated type 2 immune response following infection with Heligmosomoides polygyrus, a natural murine parasitic nematode. Elevations in cytokine gene expression and protein secretion were pronounced and comparable in inoculated B7-2-/- and B7-2+/+ mice at day 8 after H. polygyrus inoculation. However, by day 14 after infection, increases in T cell cytokine expression were markedly inhibited in H. polygyrus-inoculated B7-2-/- mice. Furthermore, elevations in serum IgE and germinal center formation were inhibited at later stages of the immune response, while elevations in serum IgG1 persisted. These findings suggest that certain T-dependent components vary in their B7-2-dependency during the type 2 immune response. They further demonstrate that B7-2 interactions are not necessary for the initiation of the type 2 immune response, but are instead required for its progression after the development of effector T cells.

Animals↗

Evolutionary alteration in anterior patterning: otx2 expression in the direct developing frog Eleutherodactylus coqui.

The homeobox gene otx2 is a key regulator for specifying the rostral part of the vertebrate head. In Xenopus, otx2 directly controls the differentiation of the cement gland, the anterior-most organ formed in the tadpole. Since embryos of a direct developing frog, Eleutherodactylus coqui, lack a cement gland, we are interested in whether altered expression of the otx2 gene is involved in this evolutionary change. We have cloned the E. coqui homologue of otx2, Ecotx2. The homeodomain of the Ecotx2 protein is identical to the mouse and zebrafish Otx2 proteins and differs by a single amino acid from the Xenopus Otx2 protein. Study of the spatiotemporal expression pattern shows that Ecotx2 RNA is progressively restricted to the anterior region of the embryo during gastrulation and becomes further restricted to the future forebrain and midbrain during neural development. In Xenopus, in addition to the conserved expression in the anterior neuroectoderm, the expression in ectoderm expands more anteriorly to the cement gland primordium. This anterior expansion of otx2 expression is not found in E. coqui, correlating with the loss of a cement gland. When misexpressed in Xenopus laevis ectoderm, Ecotx2 can activate expression of the cement-gland-specific genes XCG and XAG1, indicating that the function of activating the pathway of cement gland formation is retained by the Ecotx2 protein. Our results indicate that there are modifications in the pathway of cement gland formation, upstream of otx2 expression, in the development of E. coqui.

Amino Acid Sequence↗

Characterization of beta-lactam-resistant Bacteroides fragilis isolates by use of PCR fingerprinting.

PCR fingerprinting was used for characterization of 35 beta-lactam-resistant Bacteroides fragilis strains isolated in Sweden and Hungary. Ten B. fragilis strains showed unique PCR fingerprints by use of the M13 core primer. Their main product was a DNA fragment with a length of 2000-bp which was absent in the other 25 strains and the reference strain B. fragilis ATCC 25285. The 2000-bp fragment from four imipenem-resistant strains gave rise to positive reactions in a specific PCR for detection of ccrA. Printed by the T3B primer, five B. fragilis strains, including the imipenem-resistant strains showed unique PCR fingerprints. The investigated imipenem-resistant strains produced carbapenem-hydrolysing metallo-beta-lactamases. The study indicates that the unique PCR fingerprinting profiles shown in highly beta-lactam resistant B. fragilis strains are correlated to antimicrobial resistance. The PCR fingerprinting technique is a useful tool for differentiation of Bacteroides fragilis strains with high-level beta-lactam resistance.

Journal Article↗

Effects of theophylline on plasma levels of interleukin-4, cyclic nucleotides and pulmonary functions in patients with chronic obstructive pulmonary disease.

In order to measure the plasma levels of interleukin-4 (IL-4), cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) in patients with chronic obstructive pulmonary disease (COPD) and observe the effects of oral theophylline on them, we divided 28 COPD patients into COPD experimental group and COPD control group. Plasma levels of IL-4, cAMP and cGMP as well as parameters of pulmonary function tests were measured in these 2 groups before and after 2 weeks of treatment with oral theophylline (Protheo, 400 mg, qd) or placebo. Plasma levels of IL-4 and cGMP were significantly elevated in patients with COPD as compared with normal controls (P < 0.05), while cAMP and cAMP/cGMP were significantly lower than those in controls (P < 0.01). Plasma level of IL-4 was inversely correlated with forced expiratory volume at the first second (FEV1) and with maximum expiratory flow rate at 50% of forced vital capacity (V50) (both r = -0.46, P < 0.05) while it was directly correlated with the scores of the clinical manifestations (r = 0.57, P < 0.05) in COPD patients. Two weeks after treatment with theophylline, IL-4 and cGMP in COPD experimental group were decreased significantly while cAMP and cAMP/cGMP increased significantly (P < 0.05). The change of IL-4 was inversely correlated with the changes of FEV1 and V50 (r = -0.53 and -0.54, respectively, P < 0.05). Two weeks after placebo treatment, the COPD control group did not show such changes. We are led to conclude that IL-4 might play a role in the pathogenesis of the airway inflammation and air flow limitation in COPD patients and the mechanisms of theophylline's therapeutic effects of attenuating air-flow limitation may partially depend on its anti-inflammatory effects on the airways which, in turn, is dependent on its inhibitory effects on some inflammatory mediators such as IL-4.

Aged↗

Hamstring extensibility and transverse plane knee control relationship in athletic women.

Athletic women are at particular risk for sustaining a non-contact anterior cruciate ligament (ACL) injury. The hamstrings are vital to providing dynamic knee motion control in the sagittal and transverse planes during running stance, and some have suggested this function is enhanced when they are less extensible. This study attempted to determine the correlational relationships between hamstring extensibility and transverse plane knee kinematics and from these data to describe the dynamic transverse plane knee motion control capabilities of the hamstrings. Twenty normal athletic women (mean +/- SD; aged 21 +/- 1.6 years; height 163.3 +/- 5.7 cm; weight 60.1 +/- 3.6 kg) were evaluated for active hamstring extensibility and transverse plane knee kinematic relationships during crossover-cut stance phase. Following crossover-cut training (3 weeks) using the left (preferred) lower extremity as the stance limb, hamstring extensibility was measured. Following this, subjects were fitted with 9 retroreflective markers denoting the local segmental coordinate systems (3 markers each) of the left foot, leg and thigh. Kinematic (3-dimensional, four phase-locked cameras, 200 Hz) knee and ankle data were sampled and analyzed. Descriptive statistics and Pearson correlations between hamstring extensibility and knee and ankle kinematic variables were calculated. Statistical significance was set at P < or = 0.05 with Bonferroni adjustments. Hamstring extensibility (12 degrees +/- 8 degrees terminal extension) revealed low but significant positive correlations with tibial external rotation (6 degrees +/- 10.7 degrees) at heelstrike (19.3 degrees +/- 8.5 degrees knee flexion) (r = 0.62, P = 0.004) and tibial internal rotation (-13 degrees +/- 8.4 degrees) at peak knee flexion (57.8 degrees +/- 9.3 degrees) following heelstrike (r = 0.47, P = 0.01). Increased hamstring extensibility resulted in increased tibial external rotation at heelstrike and decreased tibial internal rotation at peak knee flexion. Increased hamstring extensibility may improve knee extensor efficiency at heelstrike by enabling greater tibial external rotation and protect the ACL at peak knee flexion by decreasing the tibial internal rotation magnitude.

Adult↗

[Gene therapy of AIDS].

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Acquired Immunodeficiency Syndrome↗

[DNA vaccination of the induction of immune responses by codelivery of IL-12 expression vector with hepatitis C structural antigens].

OBJECTIVE: In an attempt to demonstrate the utility of DNA vaccines for the tailored methods, the efficacy of immune responses would be enhanced and the types of immune responses shifted. METHODS: Four recombinant plasmids were constructed. These included the HCV coding regions for the core protein(pC) and for the core E1 and E2 together(pCE1E2) IL-12 p35 and p40. These plasmids were transfected into mammalian cells to test their protein expression and were injected into the quadriceps muscles of BALB/C mice to measure specific antibodies and cytotoxic T-lymphocyte responses. RESULTS: All the recombinant plasmids were shown to express specific antigens in cells transiently and stably. Codelivery of pIL-12 expression cassettes with pC and pCE1E2 in mice resulted in splenomegaly and the increasing number of the splenocytes. It also resulted in the enhancement of Ag-dependent CTL responses and the reduction of specific Ab response. The CTL activity were pC = 18.65% +/- 5.71%, pCE1E2 = 20.07% +/- 11.11%, pC + pIL-12 = 60.11% +/- 17.37%, pCE1E2 + pIL-12 = 67.48% +/- 15.57%, respectively. The Anti-HCV activity were pC = 0.415 +/- 0.127, pCE1E2 = 0.358 +/- 0.096, pC + pIL-12 = 0.210 +/- 0.086, pCE1E2 + pIL-12 = 0.258 +/- 0.125, respectively. CONCLUSION: Codelivery of pIL-12 with plasmid DNA can enhance the efficacy of immune responses and shift the type of immune responses. This work demonstrates the power of DNA delivery in vivo for both the production of a new generation of more effective and targeted vaccines or immuno-therapies as well as an analytic tool for the molecular dissection of the mechanisms of immune function.

Animals↗

[A study of the relationship between dynamic blood rheologic changes and the fluid resuscitation in burn patients].

OBJECTIVE: To further study the relationship between hemor rheologic changes and fluid resuscitation in patients with burn and treatment of anemia after burns. METHODS: 238 case of burn patients were divided into four groups. In which patients with major burn were allocated to group C and D according to different methods of fluid resuscitation and treatment of anemia. RESULTS: Abnormal hemorrheologic parameters could be corrected when transfusion of fresh whole blood and traditional herb medicines were given within 24-48 hours after burn injury. CONCLUSION: Dynamic observation of hemorrheologic changes is one of important index in monitoring burn shock and guiding fluid resuscitation. Blood transfusion and traditional medicine can improve abnormal blood rheologic parameters, effectively correct anemia when given early.

Adolescent↗

[A new osmotolerant and glycerol-highly-producing species--Candida glycerolgenesis Zhuge sp. nov].

The strain WL2002-5 isolated from natural samples and overproduced glycerol from glucose was identified systematically. The WL2002-5 fermented glucose and weakly fermented sucrose; assimilated glucose, sucrose, ethanol, weakly assimilated glycerol and citrate, did not assimilated nitrate, erythritol, arabinitol, mannitol. It grew well on the slopes containing 500 g/L of glucose or 10 mL/L acetate or at the temperature up to 40 degrees C. The minimum water activity for its growth was 0.890. It had a negative reaction with DBB and did vegetative reproduction by budding and easily formed Candida-type pseudohyphae but had no sexual reproduction. The molecular weight of its mitochondrial DNA was 20 kb. We conclude that WL2002-5 is a new species of Candida and nominated it Candida glycerolgenesis Zhuge sp. nov..

Candida↗

[Chemical constituents of Scolopendra negrocapitis].

The chemical constituent of Scolopendra negrocapitis Zhang et Wang were systematically analyzed and compared with S. subspinipes mutilans L. Koch. The results showed the lipid content was 18.7%, protein was 63.4% and total amino acid was 11.9%; S. negrocapitis contained the same twelve fat acids (the content of unsaturated fat acid was nearly 64% in its fat acids), twenty one amino acids and twelve trace elements as S. subspinipes mutilans had. The protein of S. negrocapitis showed forteen bands as determined by polyacrylamide gel electrophoresis. S. negrocapitis and S. subspinipes mutilans have the same main chemical composition. The contents of compositions are different. S. negrocapitis is worth as the nature resources of medicinal centepede.

Amino Acids↗

The Effect of Surfactant Monolayers on the Heat Transfer Through Air/Water and Oil/Water Interfaces Using IR Imaging Technique.

An experimental investigation on the effect of surfactant monolayers on the heat transfer through air/water and oil/water interfaces was carried out by observing the changes of surface temperature with IR Imaging Radiometer (Model 760). The heat transfer resistance of various single component and mixed monolayers at air/water and oil/water systems was studied. The results show that the surfactant monolayers introduce a noticeable heat transfer resistance to the heat transfer process across the interface. The solid monolayers exhibit lower resistance to heat transfer than the liquid monolayers at the oil/water interface. At air/water interface, the presence of monolayer decreases the evaporative cooling process and therefore increases the surface temperature rapidly. However, the presence of a monolayer at oil/water interface increases the heat transfer resistance across the oil/water interface. Heat transfer resistance increases as the chain length of fatty acid increases at the oil/water interface. The effects of phase-transition from a two-dimensional solid to the liquid state in cholesterol-arachidyl mixed monolayers was observed from the change in heat transfer resistance of the monolayers at the oil/water interface. The optimum molecular packing at the 1:3 molecular ratio in mixed surfactant monolayers of oleic acid-cholesterol and stearic acid-stearyl alcohol at the oil/water interface was also observed by this technique. Copyright 1998 Academic Press.

Journal Article↗

Transplantation of multipotent progenitors from the adult olfactory epithelium.

Mammalian olfactory epithelium produces new neurons rapidly throughout adulthood. Here, we demonstrate that precursor cells harvested from the adult olfactory epithelium, when transplanted into the nasal mucosa of host rats exposed previously to an olfactotoxic gas, engraft and participate in neuroepithelial reconstitution. In contrast to their normal neuronal fate in situ, grafted precursors harvested from bulbectomized donors produced non-neuronal cells as well as neurons. These results demonstrate that epithelial precursors activated following olfactory bulbectomy are not irreversibly committed to making neurons. Thus, olfactory progenitors are subject to a form of feedback control in vivo that regulates the types of cells that they produce within a broader-than-neuronal repertoire.

Alkaline Phosphatase↗

Prolactin activates Stat1 but does not antagonize Stat1 activation and growth inhibition by type I interferons in human breast cancer cells.

Type I interferons (IFN alpha and IFN beta) are presently used in the adjuvant treatment of several human cancers. However, these cytokines have demonstrated only modest success in breast cancer therapy, and research efforts have focused on improving their efficacy. Recent progress in understanding the molecular mechanisms underlying the antiproliferative effects of IFNs has identified the cytoplasmic transcription factor Stat1 as a critical mediator. It is, therefore, possible that IFN-induced growth inhibition of mammary epithelial cells is counteracted by other cytokines that also use Stat1. One such candidate IFN-antagonist with particular relevance to breast cancer is the mammotropic hormone prolactin (PRL). The main goal of this study was to examine whether PRL would interfere with type I IFN (IFN alpha/beta) signal transduction by competing for limited cytoplasmic Stat factors. A second aim was to test whether pretreatment of mammary tumor cell lines with IFN gamma could enhance the effect of IFN alpha/beta. By analyzing the effect of PRL on IFN alpha/beta-induced tyrosine phosphorylation of Stat proteins and their binding to IFN-regulated genes, we now report that costimulation of PRL receptors did not interfere with IFN alpha/beta signals in several human breast cancer cell lines, including T47D, MCF-7, and BT-20. Specifically, PRL did not affect IFN alpha/beta-induced tyrosine phosphorylation or heterodimerization of Stat1 and Stat2 in any cell line. Instead, IFN alpha/beta- and PRL-induced tyrosine phosphorylation of Stat1 was additive and occurred without evidence of competition for limited concentrations of cytoplasmic Stat1. A similar additive relationship was observed on IFN alpha/beta- and PRL-induced Stat3 tyrosine phosphorylation. Furthermore, electrophoretic mobility shift assays showed that type I IFNs induced predominantly Stat1-Stat2 or Stat1-Stat3 heteromeric complexes with various IFN-response elements of IFN-stimulated genes, whereas PRL induced Stat1 homodimers. Despite significant mutual use of Stats by IFNs and PRL, these results indicated a high degree of signaling specificity in the two receptor systems, and that cytoplasmic levels of Stat proteins were not limiting. Similarly, PRL did not interfere with the growth-inhibitory effect of IFN beta. On the other hand, the study indicated that pretreatment of human breast cancer cell lines with IFN gamma enhanced the growth-inhibitory action of type I IFNs, suggesting a possible avenue for improving the effect of type I IFNs in the treatment of breast cancer patients.

Animals↗

Prolonged STAT1 activation related to the growth arrest of malignant lymphoma cells by interferon-alpha.

Multiple biologic effects of interferon-alpha (IFN-alpha), including cell growth inhibition and antiviral protection, are initiated by tyrosine phosphorylation of STAT proteins. Although this signal pathway has been intensively investigated, the relevance of STAT signal persistence has received scant attention. Using paired isogenic lymphoma cells (Daudi), which either are sensitive or resistant to growth inhibition by IFN-alpha, we found comparable initial tyrosine phosphorylation of multiple STAT proteins; however, the phosphorylation durations and associated DNA-binding activities diverged. Phosphorylation and DNA-binding capacity of STAT1 decreased after 4 to 8 hours in resistant cells, as compared with 24 to 32 hours in sensitive cells, whereas phosphorylation of STAT3 and STAT5b was briefer in both lines. Functional significance of the prolonged STAT1 signal, therefore, was explored by experimental interruption of tyrosine phosphorylation, either by premature withdrawal of the IFN-alpha or deferred addition of pharmacologically diverse antagonists: staurosporine (protein kinase inhibitor), phorbol 12-myristate 13-acetate (growth promoter), or aurintricarboxylic acid (ligand competitor). Results indicated that an approximately 18-hour period of continued STAT1 phosphorylation was associated with growth arrest, but that antiviral protection developed earlier. These differences provide novel evidence of a temporal dimension to IFN-alpha signal specificity and show that duration of STAT1 activation may be a critical variable in malignant cell responsiveness to antiproliferative therapy.

Antineoplastic Agents↗