Biomedical subjects

K Zhou

Publications and source records attributed to K Zhou.

At least 37 records · Page 2Linked to original sources

PubMed2001

[Mitochondrial mechanisms of CPT-induced apoptosis in nasopharyngeal carcinoma cells].

OBJECTIVE: To understand whether CPT-induced apoptosis is related to the alteration of mitochondrial transmembrane potentials (delta psi m). METHOD: A low differentiation human nasopharyngeal carcinoma (NPC) cell line of CNE-2Z cells were used as in vitro model. After treatment with 2 mumol/L CPT for 24 h, the delta psi m was detected by the staining of rhodamine 123 (Rh123), while apoptosis was confirmed by sub-G1 cell content and the double staining of hoechst33258 and propidium iodide (PI). RESULT: CPT induced delta psi m collapse and apoptosis. With 2 mumol/L CPT treatment for 24 h, delta psi mlow cell was increased to (19.0 +/- 3.0)%, while in the DMSO control, delta psi mlow was (7.0 +/- 1.4)% P < 0.05. CONCLUSION: The disruption of the delta psi m may be one of the mechanisms of CPT-induced apoptosis in CNE-2Z cells.

Antineoplastic Agents, Phytogenic↗

PubMed2001

[Expression of the artificial cecropin CMIV-like gene of Chinese silkworm Bombyx mori in Laphygma exigua larvae].

The synthesized CMIV-like Gene was linked with the signal peptide gene of nature silkworm antibacterial peptide and was inserted into baculovirus expression vector pFastBac 1, construcing a recombinant transposing vector. The vector was transformed into DH10Bac competent E. coli cells. The recombinant Bacmid was obtained. The recombinant Bacmid was transfected into sf21 cells to get the recombinant virus. Laphygma exigua larvae were infected with the recombinant virus to express the antibacterial peptide. The hemolymph were tested to have antibacterial activity. The active antibacterial peptide was purified by acid polyacrylamide electrophoresis. The specific expression of mRNA of CMIV-like Gene was tested using Northern blotting.

Animals↗

PubMed2000

Authentication of an animal crude drug, Zaocys, by diagnostic PCR.

A pair of diagnostic primers for distinguishing the Chinese crude drug Zaocys (Zaocys dhumandes) from its substitutes was designed based on the sequence data of the original animal of the drug and substitutes. Total DNAs were extracted from genuine crude drug and 5 of its substitutes, as well as from 12 species of original animal of the snake crude drug. Diagnostic PCRs were performed using the primers with these total DNAs as a template, annealing at 60-65 degrees C. Positive amplifications were obtained from all DNA templates of Zaocys, whereas negative amplifications were obtained from that of others. The results indicate that Zaocys samples could be definitely distinguished from its substitutes by diagnostic PCR, and no incorrect discrimination was found under the same reaction conditions. The advantages of the method in the authentication of crude drugs are also discussed in the present paper.

Animals↗

PubMed2000

Fas activation reduces neutrophil adhesion to endothelial cells.

Polymorphonuclear neutrophils (PMN) express apoptotic markers and lose effector functions including adhesion, chemotaxis, and phagocytosis when cultured overnight. Although the loss of function correlates with apoptosis, it is not clear if functions are lost before an early marker of apoptosis, the display of phosphatidylserine (PS), targets PMN for removal by phagocytic cells. To address this question, freshly isolated PMN were treated with Fas-activating antibodies to induce apoptosis rapidly. Early markers of apoptosis and PMA-stimulated adhesion to endothelial cells were measured. After 1 h of Fas exposure, only 16% PMN had externalized PS. In contrast, Fas activation reduced PMA-stimulated adhesion between 68 and 27% depending on PMA concentration. The loss of adhesion was accompanied by a reduction in beta2 integrin expression and receptor clustering. These results indicate that the Fas-induced loss of adhesion may precede PS externalization and could limit participation in the inflammatory response before PS externalization targets PMN for removal.

Animals↗

PubMed2000

Sequence variations of the hypervariable region of hepatitis C virus and their clinical significance.

OBJECTIVE: To understand the clinical significance of sequence variations in the hypervariable region (HVR) of hepatitis C virus during infection. METHODS: Eight patients with acute hepatitis C and 20 patients with chronic hepatitis C were followed up for two years. Blood samples were taken at intervals of six months for analysis of HCV-HVR sequences by reverse transcription-polymerase chain reaction (RT-PCR) and direct sequencing methods. RESULTS: HCV-HVR sequences of the 28 patients changed in various degrees. 92% of these nucleotide substitutions led to changes of corresponding aminoacid sequences. Only 8% of changed nucleotide were synonymous substitutions. Of 27 amino acids variation of amino acid ranged from 1 to 20 (mean 8, 30%). The most common nucleotide substitution (62%) occurred in the first position of codon, 31% in the second and the rest in the third. HVR variation rate was 0.89 x 10(-1) per genome site per year in acute hepatitis C, compared with 2.31 x 10(-1) per genome site per year in chronic hepatitis C (P < 0.05), but had no relation to HCV subtype. Variation of HVR in the flare up type (ALT > 150 mu/L) was much more than that in the quiescent type (ALT < 100 mu/L). CONCLUSION: Our results suggested that sequence variation of HVR during HCV chronic infection seems to be an adaptive response to HCV to evade the host immune pressure and might play a major role in the establishment of persistent infection as well as in the flare-up of hepatitis.

Acute Disease↗

PubMed2000

[Application of RAPD in the taxonomy of the genus Fritillaria].

OBJECTIVE: To provide the molecular data for the right application of the Chinese medicine "Beimu". METHOD: Using the technology of RAPD, we studied the relationship among 12 samples of Beimu. RESULT: The total genomic DNA of all the samples are about 21 Kb in size. Among 20 primers used, the five primers can reatedly generate a certain specified amplified band type, 27 bands were recored from all amplified products and 25 polymorphic fragments were found in it. The size of amplified fragments is between 450 bp and 1904 bp. CONCLUSION: The similarity within species is higher than those between species. The relationship of Fritillaria anhuiensis and F. puqiensis is the farest, while F. thunbergii and F. puqiensis is the closest.

DNA Primers↗

PubMed1999

The hepatitis C virus internal ribosome entry site adopts an ion-dependent tertiary fold.

Hepatitis C virus (HCV) contains an internal ribosome entry site (IRES) located in the 5' untranslated region of the genomic RNA that drives cap-independent initiation of translation of the viral message. The approximate secondary structure and minimum functional length of the HCV IRES are known, and extensive mutagenesis has established that nearly all secondary structural domains are critical for activity. However, the presence of an IRES RNA tertiary fold and its functional relevance have not been established. Using chemical and enzymatic probes of the HCV IRES RNA in solution, we show that the IRES adopts a unique three-dimensional structure at physiological salt concentrations in the absence of additional cofactors or the translation apparatus. Folding of the IRES involves cooperative uptake of magnesium and is driven primarily by charge neutralization. This tertiary structure contains at least two independently folded regions which closely correspond to putative binding sites for the 40 S ribosomal subunit and initiation factor 3 (eIF3). Point mutations that inhibit IRES folding also inhibit its function, suggesting that the IRES tertiary structure is essential for translation initiation activity. Chemical and enzymatic probing data and small-angle X-ray scattering (SAXS) experiments in solution show that upon folding, the IRES forms an extended structure in which functionally important loops are exposed. These results suggest that the 40 S ribosomal subunit and eIF3 bind an HCV IRES that is prefolded to spatially organize recognition domains.

Base Sequence↗

PubMed1999

[Evaluation of the arterial phase of biphase enhanced SCT in the diagnosis of small HCC].

OBJECTIVE: To evaluate the value of the arterial phase (AP) of biphase enhanced SCT in the diagnosis of small HCC and investigate the criteria, initial time, ending time and duration of AP. METHODS: From May 1995 to March 1999, patients with small HCC proved by surgicopathology including 49 cases (N = 53) in the 1st group, 148 cases (N = 186) in the 2nd group and 52 cases (N = 52)n in the 3rd group were collected. Biphase dynamic enhanced SCT scans were performed in all patients of three groups and additional single-level dynamic scans only done in the 3rd group. The detectability, diagnostic accuracy of the lesions and enhancement of the lesion in AP were analyzed statistically. In addition, the initial time, ending time and duration of AP were measured. RESULTS: The results of the 1st group showed the detectability of small HCC was 88.68% in AP and 90.57% in both phases, higher than those by US. Markedly enhanced lesion in AP accounted for 76% and 78% in the 2nd and 3rd groups respectively. The initial, ending and duration times of AP measured on single-level dynamic scans were 16.9 s, 39.6 s and 22.7 s in average respectively. CONCLUSION: The biphase dynamic SCT especially its arterial phase appears to be very valuable in diagnosing small HCCs. In light of short duration of AP, understanding and strict control of AP is obviously imperative.

Adult↗

PubMed1999

[Effects of synthetic peptides on ovarian cancer cells].

OBJECTIVE: To observe the effect of follicle-stimulating hormone(FSH) synthetic peptides(FSH binding Fragment), FSH and synthetic peptides on the proliferation of human epithelial ovarian cancer cell. METHODS: Human epithelial ovarian cancer cells lines SKOV3, OVCAR, AO and 3AO were incubated with FSH, FSH binding fragment, FSH and the binding fragment respectively. The cell proliferation was detected by methyl thiazolyl tetrazolium (MTT) technique. RESULTS: The rate of proliferation in the cancer cell was increased apparently as increasing in the concentration of FSH and the rate of proliferation average value 28.0%, and was decreased apparently as increasing in the concentration of the synthetic peptides and the rate of inhibition average value 8.3%. When the cell was expressed in FSH, the proliferation was decreased apparently as increasing in the concentration of the peptides and the rate of inhibition average valve 3.1%. CONCLUSIONS: It is suggested that FSH binding fragment can inhibit the proliferation of ovarian cancer cell. The FSH binding fragment could be used as a binding part of anticancerous complex for ovarian cancer.

Dose-Response Relationship, Drug↗

PubMed1999

[Clinical study on effect of shenqi fuzheng injection combined with chemotherapy in treating gastric cancer].

OBJECTIVE: To assess the clinical effect of Shenqi Fuzheng Injection (SQFZI) combined with chemotherapy in treating gastric cancer. METHODS: One hundred and twenty patients were randomly divided into 3 groups, the combined therapy group (Group A), the chemotherapy group (Group B) and the SQFZI group (Group C) and the effect on remission and stabilization of patients were observed. RESULTS: The remission rate and stabilizing rate of Group A were 16.1% and 87.1%, those of Group B were 13.5% and 64.9%, the difference between the two groups was significant, P < 0.05. The symptom and living quality improving rate of Group A were 75.8% and 43.5% respectively, those of Group C were 61.9% and 57.1% and of Group B were 35.1% and 29.7% respectively. SQFZI showed good protective effect on hemopoietic system, 4.8% patients in Group A with WBC count lower than 4 x 10(9)/L, while the percentage reached 21.6% in Group B, the difference between the two groups was significant, P < 0.05. Moreover, SQFZI could raise activity of NK cell, macrophage and T-lymphocyte subgroups, without any injury on heart, liver and kidney function or other adverse reaction. CONCLUSION: SQFZI combined with chemotherapy has certain effect of remission and stabilization on gastric cancer, the clinical effect is significant to patients with Qi-Deficiency, and is reliable and safe.

Adult↗

PubMed1998

A novel, putative MEK kinase controls developmental timing and spatial patterning in Dictyostelium and is regulated by ubiquitin-mediated protein degradation.

We have identified a developmentally regulated, putative MEK kinase (MEKKalpha) that contains an F-box and WD40 repeats and plays a complex role in regulating cell-type differentiation and spatial patterning. Cells deficient in MEKKalpha develop precociously and exhibit abnormal cell-type patterning with an increase in one of the prestalk compartments (pstO), a concomitant reduction in the prespore domain, and a loss of the sharp compartment boundaries, resulting in overlapping prestalk and prespore domains. Overexpression of MEKKalpha or MEKKalpha lacking the WD40 repeats results in very delayed development and a severe loss of compartment boundaries. Prespore and prestalk cells are interspersed throughout the slug. Analysis of chimeric organisms suggests that MEKKalpha function is required for the proper induction and maintenance of prespore cell differentiation. We show that the WD40 repeats target MEKKalpha to the cortical region of the cell, whereas the F-box/WD40 repeats direct ubiquitin-mediated MEKKalpha degradation. We identify a UBC and a UBP (ubiquitin hydrolase) that interact with the F-box/WD40 repeats. Our findings indicate that cells lacking the ubiquitin hydrolase have phenotypes similar to those of MEKKalpha null (mekkalpha-) cells, further supporting a direct genetic and biochemical interaction between MEKKalpha, the UBC, and the UBP. We demonstrate that UBC and UBP differentially control MEKKalpha ubiquitination/deubiquitination and degradation through the F-box/WD40 repeats in a cell-type-specific and temporally regulated manner. Our results represent a novel mechanism that includes targeted protein degradation by which MAP kinase cascade components can be controlled. More importantly, our findings suggest a new paradigm of spatial and temporal control of the kinase activity controlling spatial patterning during multicellular development, which parallels the temporally regulated degradation of proteins required for cell-cycle progression.

Amino Acid Sequence↗

PubMed1998

Crystal structure of a hepatitis delta virus ribozyme.

The self-cleaving ribozyme of the hepatitis delta virus (HDV) is the only catalytic RNA known to be required for the viability of a human pathogen. We obtained crystals of a 72-nucleotide, self-cleaved form of the genomic HDV ribozyme that diffract X-rays to 2.3 A resolution by engineering the RNA to bind a small, basic protein without affecting ribozyme activity. The co-crystal structure shows that the compact catalytic core comprises five helical segments connected as an intricate nested double pseudoknot. The 5'-hydroxyl leaving group resulting from the self-scission reaction is buried deep within an active-site cleft produced by juxtaposition of the helices and five strand-crossovers, and is surrounded by biochemically important backbone and base functional groups in a manner reminiscent of protein enzymes.

Base Sequence↗

PubMed1998

Guanine nucleotide exchange factors regulate specificity of downstream signaling from Rac and Cdc42.

The Rac and Cdc42 GTPases regulate diverse cellular behaviors involving the actin cytoskeleton, gene transcription, and the activity of multiple protein and lipid kinases. All of these pathways can potentially become activated when GTP-Rac or GTP-Cdc42 is formed in response to external cell signals, yet it is evident that each activity must also be able to be controlled individually. The mechanisms by which such specificity of GTPase signaling in response to upstream stimuli is achieved remains unclear. We investigated the action of several well characterized guanine nucleotide exchange factors (GEFRho) to activate Rac- and/or Cdc42-dependent kinase pathways. Coexpression studies in COS-7 cells revealed that the ability of individual guanine nucleotide exchange factors (GEFs) to activate the p21-activated kinase PAK1 could be dissociated from activation of c-Jun amino-terminal kinase, even though activation of both pathways requires the action of the GEFs on Rac and/or Cdc42. In contrast, expression of constitutively active forms of Rac or Cdc42 effectively stimulated both downstream kinases. We conclude that GEFs can be important determinants of downstream signaling specificity for members of the Rho GTPase family.

Animals↗

PubMed1998

A general module for RNA crystallization.

Crystallization of RNA molecules other than simple oligonucleotide duplexes remains a challenging step in structure determination by X-ray crystallography. Subjecting biochemically, covalently and conformationally homogeneous target molecules to an exhaustive array of crystallization conditions is often insufficient to yield crystals large enough for X-ray data collection. Even when large RNA crystals are obtained, they often do not diffract X-rays to resolutions that would lead to biochemically informative structures. We reasoned that a well-folded RNA molecule would typically present a largely undifferentiated molecular surface dominated by the phosphate backbone. During crystal nucleation and growth, this might result in neighboring molecules packing subtly out of register, leading to premature crystal growth cessation and disorder. To overcome this problem, we have developed a crystallization module consisting of a normally intramolecular RNA-RNA interaction that is recruited to make an intermolecular crystal contact. The target RNA molecule is engineered to contain this module at sites that do not affect biochemical activity. The presence of the crystallization module appears to drive crystal growth, in the course of which other, non-designed contacts are made. We have employed the GAAA tetraloop/tetraloop receptor interaction successfully to crystallize numerous group II intron domain 5-domain 6, and hepatitis delta virus (HDV) ribozyme RNA constructs. The use of the module allows facile growth of large crystals, making it practical to screen a large number of crystal forms for favorable diffraction properties. The method has led to group II intron domain crystals that diffract X-radiation to 3.5 A resolution.

Base Sequence↗

PubMed1998

Disruption of Dictyostelium PI3K genes reduces [32P]phosphatidylinositol 3,4 bisphosphate and [32P]phosphatidylinositol trisphosphate levels, alters F-actin distribution and impairs pinocytosis.

To understand how phosphatidylinositol 3-kinase (PI3K) modulates cell structure and function, we examined the molecular and cellular defects of a Dictyostelium mutant strain (pik1(Delta)2(Delta)) missing two (DdPIK1 and 2) of three PI3K genes, which are homologues of the mammalian p110 subunit. Levels of [32P]phosphatidylinositol 3, 4 bisphosphate (PI(3,4)P2) and [32P]phosphatidylinositol trisphosphate (PIP3) were reduced in pik1(Delta)2(Delta), which had major defects in morphological and functional correlates of macropinocytosis. This was accompanied by dramatic deficits in a subset of F-actin-enriched structures such as circular ruffles, actin crowns and pseudopodia. Although pik1(Delta)2(Delta) were mobile, they failed to aggregate into streams. Therefore we conclude that PIK1 and 2, possibly through modulation of the levels of PIP3 and PI(3,4)P2, regulate the organization of actin filaments necessary for circular ruffling during macropinocytosis, the extension of pseudopodia and the aggregation of cells into streams, but not the regulation of cell motility.

Actins↗

PubMed1998

[A study on enzymatic activities of bronchoalveolar lavage fluid in patients with interstitial lung diseases].

OBJECTIVE: To evaluate the relationship between enzymatic activities in bronchoalveolar lavage fluid(BALF) and interstitial lung diseases(ILDs). METHOD: Cellular components and levels of superoxide dismutase (SOD), glutathione peroxidase(GSH-PX), angiotensin converting enzyme(ACE) and lactate dehydrogenase(LDH) in BALF in 30 cases of ILDs: including 18 patients with idiopathic pulmonary fibrosis (IPF) and 12 patients with sarcoidosis (Sarc) and 9 healthy controls were determined. RESULT: (1) The levels of BALF-SOD, GSH-PX were significantly decreased(P < 0.01), and BALF-ACE, LDH were markedly increased (P < 0.05) in patients with IPF, and BALF-ACE were also increased in patients with Sarc(P < 0.05); (2) There were good correlation between ACE and percentage of lymphocyte (r1 = 0.6574, P < 0.05), and ratio of CD4+/CD8+ (r2 = 0.9544, P < 0.001) in BALF of Sarc group. CONCLUSION: Determining enzymatic activities could be helpful to study pathogenesis and diagnosis of ILDs, BALF-ACE may be as a good marker of disease activity of Sarcoidosis.

Adult↗

PubMed1998

[Rapid determination of creatinine in urine by capillary electrophoresis in a multiple injection mode].

The level of urinary creatinine in clinic diagnosis is one of the useful index of renal function, and is often used as a reference of some biochemical substance determined in random urine. In this paper, a rapid multiple-injection capillary electrophoresis(CE) method for the determination of urinary creatinine is reported. A 36 cm x 50 microns coated capillary, a phosphoric acid buffer(0.1 mol/L, pH 2.5)and UV-detector at 200 nm wavelength were used in this method. In comparing with single injection for five samples, the analysis time of five successive injections can save 20 min. By use of pyridine as internal standard, the correlation between the concentrations of creatinine and the corresponding peak height ratios of creatinine and pyridine is good(r = 0.9996), and the assay precision is acceptable. We also compared the CE method with the bio-analyser(Jeffe' kinetic), and the results showed a satisfactory correlation (r = 0.9773, n = 12) and a shorter analysis time.

Creatinine↗

PubMed1998

[Effect of hyperbaric oxygen on cerebral microcirculation and tissue cells in animals with cerebral ischemic injury].

Changes of cerebral microcirculation and tissue cells after Hyperbaric Oxygen (HBO) exposure were observed in 136 gerbils with cerebral ischemia by observation of meningeal microcirculation pathological study in cerebral tissues and determination of Na, K-ATPase. It is indicated that HBO may be helpful in improving microcirculatory dynamics and other microcirculatory functions, and enhancing cerebral tissue cell activity and cell function, as well as increasing oxygen content. It is suggested that HBO (250 approximately 300kPa) may play a role in protecting vessel endothelial cells and nerve cells.

Adenosine Triphosphatases↗