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Biomedical subjects

J Zeng

Publications and source records attributed to J Zeng.

At least 127 records · Page 7Linked to original sources

ATPase activities of rabbit and bovine lens epithelial microsomes: a continuous fluorimetric assay study.

Purine nucleoside phosphorylase (EC 2.4.2.1) catalyzes the irreversible phosphorolysis of 7-methylguanosine (m7Guo), a fluorescent guanosine analogue. Using purine nucleoside phosphorylase and m7Guo, a continuous fluorimetric assay for microsomal ATPases from rabbit and bovine lens is described. The decrease in m7Guo fluorescence intensity at 380 nm, which represents the hydrolysis of ATP, is linear with time up to exhausting all m7Guo. The rate of the fluorescence decrease depends on the sample protein concentration. In the presence of ATPase inhibitors, ion-specific ATPase activities in the lens were determined from the difference of the fluorescence decay rates. Using the fluorimetric assay thapsigargin-sensitive Ca-ATPase in the bovine lens epithelium has been characterized. The fluorimetric assay provides a number of advantages over previous membrane ATPase assays.

Adenosine Triphosphate↗

[Measurement of the reserve function of inspiratory muscle and its clinical significance].

The principles of measuring inspiratory muscle tension-time index (TTim) and the ratio of the works of inspiration over the maximal works of inspiration (Wi/Wi(max)) were investigated and their formulae were deduced, i.e. TTim = (Pi x Ti)/(MIP x Ttot) and Wi/Wi(max) = (Pi x VT)/(MIP x IC). The importance of the inspiratory pressure and the maximal inspiratory pressure (MIP) measured at function residual capacity (FRC) level was emphasized. Both TTim and Wi/Wi(max) were measured in 35 normal subjects and 89 patients with chronic obstructive pulmonary disease (COPD). The results showed that normal value of TTim was 0.0253 +/- 0.0055 which corresponds to the normal value of the diaphragm tension-time index (TTdi = 0.02-0.03) reported by Bellemare. Patients with COPD had a mean TTim much higher than that of normals (P < 0.01). The works of inspiration (Wi) in patients with COPD increased, while the maximal works of inspiration (Wi(max)) declined, so Wi/Wi(max) became significantly greater than that of normals (P < 0.01). The results also showed that there was a linear relationship between Pi/Pimax and TTim or Wi/Wimax (r = 0.7891, 0.9738, 0.6459, 0.9327, P < 0.01). Therefore, we suggest that both TTim and Wi/Wimax can be used as clinical indices to reflect the reserve function of inspiratory muscles.

Adult↗

Chinese character operating system of traditional Chinese medicine and pharmacology (TCMP).

With the development in research, teaching and literature work in traditional Chinese medicine and pharmacology (TCMP) by means of computers, it has been found that the existing Chinese character operative systems cannot meet the need of carrying out information processing and software development in this field, since these systems do not include many of the common and special terms in TCMP. This makes it inconvenient to exchange academic thoughts in information processing in this field with our colleagues at home and abroad and greatly affects the sharing of the literature data in TCMP. It is therefore necessary to develop a Chinese character operating system applicable to the use of computers in the research of TCMP. Recently, we have developed jointly a Chinese character operating system of TCMP. This system is based on the original GB2312-80 Chinese character international code, to which are added 1,150 Chinese characters commonly used in TCMP. The five-stroke code, which make input possible according to word forms, are used for the input of expanded words. Besides, the system also provides the codes of a number of common names of Chinese materia medica, acupoint names, common terms in traditional Chinese medicine (TCM), TCM disease names, names of classic TCM works, etc. It also provides a convenient character-creating software. We hope that our work will lead to discussions concerning the difficult problems in computer processing of TCMP literature, i.e. the Chinese character.

Database Management Systems↗

[A study on the simultaneous HPLC determination of chlorhexidine and its impurity 4-chloroaniline].

This paper reports a new RP-HPLC method using phenacetin internal standard (IS) for simultaneous determination of chlorhexidine (CH) and its impurity 4-chloroaniline (4-CA). We use the LC-3A HPLC instrument, a Zorbax-C8 column (25cm x 4.6mm id, 10 microns), a variable wave length UV detector and C-RIA chromatographic data processor. A mixture of methanol and 0.2 mol/L NaH2PO4 (50.5 : 49.5, pH 3.0) is used as mobile phase, whose flow rate is 1.0 ml/min. Sample solution of 15 microliters is injected onto the column, which is maintained at 30 degrees C. The eluent is monitored at 240nm. The retention times are 16.72 min for CH and 5.54 min for 4-CA. The standard curves of CH and 4-CA are established using peak area ratio of CH/IS and 4-CA/IS to concentration respectively. They are both linear (r = 0.9999) within 15-200 micrograms/ml for CH and 60-1200 micrograms/ml for 4-CH. The limits of detection are 1.33 micrograms/ml for CH AND 7.5 micrograms/ml for 4-CA (S/N = 3). The analytical recovery rates are 100.1% for CH and 103.1% for 4-CA. This method has been used for investigating the stability of CH preparation.

Aniline Compounds↗

Differential cDNA cloning by enzymatic degrading subtraction (EDS).

We describe a new method, called enzymatic degrading subtraction (EDS), for the construction of subtractive libraries from PCR amplified cDNA. The novel features of this method are that i) the tester DNA is blocked by thionucleotide incorporation; ii) the rate of hybridization is accelerated by phenol-emulsion reassociation; and iii) the driver cDNA and hybrid molecules are enzymatically removed by digestion with exonucleases III and VII rather than by physical partitioning. We demonstrate the utility of EDS by constructing a subtractive library enriched for cDNAs expressed in adult but not in embryonic rat brains.

Animals↗

Oligosaccharides from human milk as revealed by matrix-assisted laser desorption/ionization mass spectrometry.

In this study neutral and acidic oligosaccharide fractions prepared from human milk have been investigated using matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). The fraction of neutral oligosaccharides was separated by gel permeation chromatography (GPC) and the resulting subfractions were analyzed by MALDI-MS using the positive ion mode. Several low-molecular-weight glycans (degree of polymerization up to 13) were observed whose structures have already been elucidated. In addition, a variety of so far unknown large-sized carbohydrates was detected whose molecular weights range from M(r) 2242 to 8000. The large-sized glycans which possess a low abundance appear to be composed of both lactosamine and fucose residues attached to the lactose unit at the reducing end of the sugar chains with a highly variable stochiometry. Following subfractionation by GPC, acidic (i.e., containing sialic acid) glycans were analyzed by MALDI-MS using both positive and negative ion mode. Because of the inferior stability of acidic glycans, various matrices were applied and compared with respect to signal intensity, resolution, and analyte stability.

Chromatography, Gel↗

Identification and zinc dependence of the bovine herpesvirus 1 transactivator protein BICP0.

Bovine herpesvirus 1 (BHV-1) specifies and unspliced early 2.6-kb RNA (ER2.6) which is 3' coterminal with exon 2 of the 2.9-kb immediate-early (IE) RNA. The two transcripts have a common open reading frame (676 codons). The predicted protein, designated BHV-1 infected cell protein 0 (BICP0), contains a zinc finger domain with homology to ICP0 of herpes simplex virus type 1 and protein 61 of varicella-zoster virus, and depending on the promoter, it acts as a strong activator or as a repressor in transient expression assays. In situ immunoadsorbent assays using antisera against synthetic oligopeptides demonstrated that BICP0 accumulates in nuclei of BHV-1-infected cells, as expected for an IE gene product involved in gene regulation. Western blots (immunoblots) revealed a BHV-1-specific 97-kDa protein which was detectable during the IE phase and also at later periods of infection, indicating that the kinetics of BICP0 synthesis is consistent with the switch from IER2.9 to ER2.6. To confirm that ER2.6 encoded the 97-kDa BICP0 protein, a DNA fragment containing BICP0-coding sequences was inserted into the Autographa californica baculovirus genome. A recombinant protein, identified by its reactivity with antipeptide sera, exhibited the same electrophoretic mobility as BICP0 specified by BHV-1. We microinjected Xenopus oocytes with a BICP0 effector plasmid and a promoter-chloramphenicol acetyltransferase plasmid. BICP0-induced stimulation of this promoter was strongly reduced when intracellular zinc was chelated by thionein, indicating that the effect of BICP0 is zinc dependent.

Amino Acid Sequence↗

[Expression of human chorionic gonadotropin, human placental lactogen and pregnancy-specific 1-glycoprotein in malignant trophoblastic neoplasms].

The expression of placental hormones in 91 malignant trophoblastic neoplasms was studied immunohistochemically using a panel of antibodies against hCG, human placental lactogen (hPL) and pregnancy-specific 1-glycoprotein (SP). The results indicated that the expression of hCG in invasive moles was weaker than that in choriocarcinoma, but the expression of hPL and SP1 was stronger than those in choriocarcinoma. The expression of hPL and SP1 in the metastatic tumors of invasive moles was weaker than the expression at in the primary tumors, but the secretory capacity of hCG in metastatic choriocarcinomas was stronger than that in the primary neoplasms. In this study, the qualities of expression of the placental hormones in invasive moles and choriocarcinomas corresponded to the degree of tumor malignancy, the biological behaviour and the grading of trophoblastic cell differentiation. We believe that the detection of hCG, hPL and SP1, in malignant trophoblastic neoplasms was of value for establishing tumor diagnosis and typing and for judgement on prognosis.

Biomarkers, Tumor↗

[A genetic study of adult polycystic kidney disease].

The 3'HVR has been considered as the most useful probe in making the linkage analysis of adult polycystic kidney disease (APKD). We examined three APKD families using this probe. Their polymorphism information was then computed for linkage study using LINKAGE package and computer programme HOMOG for homogeneity test. One family was defined as non PKD 1, because the disease locus was failed to show linkage to the 3'HVR marker. Of the other two families, one can still not be identified as non PKD1, but had distinguishable meiotic recombinations, another was in favour of linkage to 3'HVR and therefore judged to be PKD1. The presence of genetic heterogeneity could significantly reduce the value of linked probe analysis as a tool for gene diagnosis of APKD.

Adolescent↗

[Effect of indirect fluorescent antibody test for the diagnosis of amoebiasis].

Indirect fluorescent antibody test (IFAT) was performed by the method of [symbol: see text] (1971) with some slight modifications for the diagnosis of amoebiasis. The antigen used was prepared from 48-hour culture of Entamoeba histolytica (strains G2, G3, and G5) in LAS diphasic medium and only those amoebae clinging the culture were collected. The serum samples or blood drops on filter paper were positive by the improved IFAT in all of the 54 cases of amoebic liver abscess and the positive reactions of most cases were very marked. Of 23 cases of acute amoebic dysentery, 19 were positive, the positive rate being 82.6%. The results also show that 42 cases of other diseases and 40 healthy persons were all negative.

Animals↗

Effects of thymosin on the secretion of the sexual hormone in rats with experimental varicocele.

Hormonal abnormality in experimental varicocele induced by partial ligation of left renal vein and the effect of thymosin (fraction 5) on the secretion of the sexual hormone were studied in rats. Serum testosterone level was lowered after varicocele formation, but could be reversed or increased by intraperitoneal administration of thymosin 5 mg/rat twice a week for 4 weeks with concomitant increase of testicular testosterone content. Changes in FSH and LH both in serum and testis, determined with immunoassay method as that of testosterone, were insignificant as compared with control or thymosin treated groups.

Animals↗

Influence of thymosin on the level of cyclic nucleotides in rats with experimental varicocele.

Experimental varicocele was induced by partial ligation of the left renal vein in rats, and the effect of thymosin (fraction 5), a hormone secreted by the thymus, on the level of cyclic nucleotides was investigated. The levels of cAMP and cGMP in plasma and testicular tissue were estimated using radioimmunoassay techniques. The results showed that the level of cAMP in bilateral testicular tissues in rats with experimental varicocele decreased considerably, especially in the left testicular tissue as compared with that in the control group (P < 0.05). 4 weeks after thymosin administration (0.5 mg/rat, twice a week), the level of cAMP in bilateral testicular tissues rose significantly, as compared with that in the experimental varicocele group (P < 0.01). These findings indicate that there exists imbalance in the metabolism of cyclic nucleotides in rats with experimental varicocele. This suggests that the abnormal level of cyclic nucleotides is one of the causes of infertility with varicocele. Since thymosin participates in the metabolism and regulation of cyclic nucleotides, it may have a direct or an indirect effect on the growth and function of reproductive cells.

Animals↗

Effects of alcohol-induced lipid interdigitation on proton permeability in L-alpha-dipalmitoylphosphatidylcholine vesicles.

6-Carboxyfluorescein was employed to examine the effect of alcohol-induced lipid interdigitation on proton permeability in L-alpha-dipalmitoylphosphatidylcholine (DPPC) large unilamellar vesicles. Proton permeability was measured by monitoring the decrease of 6-carboxyfluorescein fluorescence after a pH gradient from 3.5 (outside the vesicle) to 8.0 (inside the vesicle) was established. At 20 degrees C and below 1.2 M ethanol, the fluorescence decrease is best described by a single exponential function. Above 1.2 M ethanol, the intensity decrease is better described by a two-exponential decay law. Using the fitted rate constants and the vesicle radii determined from light-scattering measurements, the proton permeability coefficient, P, in DPPC vesicles was calculated as a function of ethanol concentration. At 20 degrees C, P increases monotonically with increasing ethanol content up to 1.0 M, followed by an abrupt increase at 1.2 M. The vesicle size also exhibits a sudden increase at around 1.2 M ethanol, which has been shown to result from vesicle aggregation rather than vesicle fusion. The abrupt increases in P and in vesicle size occur at the concentration region close to the critical ethanol concentration for the formation of the fully interdigitated gel state of DPPC. At 14 degrees C, the abrupt change in P shifts to 1.9-2.0 M ethanol, completely in accordance with the ethanol-temperature phase diagram of interdigitated DPPC. Effects of methanol and benzyl alcohol on lipid interdigitation have also been examined. At 20 degrees C, DPPC large unilamellar vesicles exhibit a dramatic change in P at 3 M methanol and at 40 mM benzyl alcohol. These concentrations come close to the critical methanol and benzyl alcohol concentrations for the formation of fully interdigitated DPPC structures determined previously by others. It can be concluded that proton permeability increases dramatically as DPPC is transformed from the noninterdigitated gel to the fully interdigitated gel state by high concentrations of alcohol. This marked increase in proton permeability can be attributed to the combined effect of the changes in membrane thickness and surface charge density, due to the ethanol-induced lipid interdigitation. The possible effects of the increased proton permeability caused by ingested ethanol on gastric mucosal membranes are discussed.

1,2-Dipalmitoylphosphatidylcholine↗

[Determination of ceftizoxime in serum by high performance liquid chromatography].

This paper reports the determination of ceftizoxime, a new cephalosporin antibiotic, in serum by RP-HPLC. The technique for serum protein precipitation by adding perchloric acid is adapted for sample preparations. After centrifugation, a 100 microliters portion of aqueous phase is injected into chromatographic column of ultrasphere CN, the mobile phase being a mixture of methanol and 1% acetic acid (15:85). The standard curve is linear within the range of 2.5-120 mg/L in serum. The detection limit of ceftizoxime in serum stands at 0.1 mg/L. The recoveries from serum samples after protein precipitation reach 95.4-96.3%. The within day CVs and interday CVs are 2.7-3.3% and 4.8-7.6%, respectively. This method is rapid, reliable and reproducible. It has been utilized to measure the concentration of ceftizoxime in patient serum for clinical research and pharmacokinetic studies.

Ceftizoxime↗

[The use of domestic viscoelastic substances in IOL implantation and their toxicity to endothelial cells in vitro].

34 senile cataract patients were randomly divided into groups of 1% sodium hyaluronate (Healon) 14 patients, 2% hydroxypropylmethylcellulose (HPMC) 10 patients and Viscose 10 patients for extracapsular cataract extraction with IOL implantation. The rate of endothelial cell loss, the corneal thickness, the intraocular pressure, and the anterior chamber reactions were investigated; the toxicity of the viscoelastic substances on human corneal endothelial cells was studied in vitro for comparative evaluation.

Cataract↗

X-ray crystal structure of canine myeloperoxidase at 3 A resolution.

The three-dimensional structure of the enzyme myeloperoxidase has been determined by X-ray crystallography to 3 A resolution. Two heavy atom derivatives were used to phase an initial multiple isomorphous replacement map that was subsequently improved by solvent flattening and non-crystallographic symmetry averaging. Crystallographic refinement gave a final model with an R-factor of 0.257. The root-mean-square deviations from ideality for bond lengths and angles were 0.011 A and 3.8 degrees. Two, apparently identical, halves of the molecule are related by local dyad and covalently linked by a single disulfide bridge. Each half-molecule consists of two polypeptide chains of 108 and 466 amino acid residues, a heme prosthetic group, a bound calcium ion and at least three sites of asparagine-linked glycosylation. There are six additional intra-chain disulfide bonds, five in the large polypeptide and one in the small. A central core region that includes the heme binding site is composed of five alpha-helices. Regions of the larger polypeptide surrounding this core are organized into locally folded domains in which the secondary structure is predominantly alpha-helical with very little organized beta-sheet. A proximal ligand to the heme iron atom has been identified as histidine 336, which is in turn hydrogen-bonded to asparagine 421. On the distal side of the heme, histidine 95 and arginine 239 are likely to participate directly in the catalytic mechanism, in a manner analogous to the distal histidine and arginine of the non-homologous enzyme cytochrome c peroxidase. The site of the covalent linkage to the heme has been tentatively identified as glutamate 242, although the chemical nature of the link remains uncertain. The calcium binding site has been located in a loop comprising residues 168 to 174 together with aspartate 96. Myeloperoxidase is a member of a family of homologous mammalian peroxidases that includes thyroid peroxidase, eosinophil peroxidase and lactoperoxidase. The heme environment, defined by our model for myeloperoxidase, appears to be highly conserved in these four mammalian peroxidases. Furthermore, the conservation of all 12 cysteine residues involved in the six intra-chain disulfide bonds and the calcium binding loop suggests that the three-dimensional structures of members of this gene family are likely to be quite similar.

Amino Acid Sequence↗

Conformational analysis of the anomeric forms of kojibiose, nigerose, and maltose using MM3.

Energy surfaces were computed for relative orientations of the relaxed pyranosyl rings of the two anomeric forms of kojibiose, nigerose, and maltose, the (1----2)-alpha, (1----3)-alpha, and (1----4)-alpha-linked D-glucosyl disaccharides, respectively. Twenty-four combinations of starting conformations of the rotatable side-groups were considered for each disaccharide. Optimized structures were calculated using MM3 on a 20 degree grid spacing of the torsional angles about the glycosidic bonds. The energy surfaces of the six disaccharides were similar in many respects but differed in detail within the low-energy regions. The maps also illustrate the importance of the exo-anomeric effect and linkage type in determining the conformational flexibility of disaccharides. Torsional conformations of known crystal structures of maltosyl-containing molecules lie in a lower MM3 energy range than previously reported.

Carbohydrate Conformation↗

Composition of human islet cell preparations for transplantation.

To study the cellular composition of human islet cell isolates for transplantation, formalin-fixed and paraffin-embedded cell pellets were stained by the immunoperoxidase method with a panel of antibodies characterising endocrine, epithelial, soft tissue and haematolymphoid components. Immediately after separation, the isolates contained 30-80% islet cells, differing mainly in the content of islet and acinar cells, whereas the soft tissue, ductal/ductular and haematolymphoid elements comprised a relatively constant 10-20%. After 1 week in culture the islet cell content of less highly purified isolates (30-40% islets) dropped dramatically to 5%. The highly purified isolates (70-80% islets) showed only a minimal change in cellular composition; however, approximately two-thirds of islet cells were degranulated and did not stain for insulin. Haematolymphoid components were still present in all cultured isolates. We conclude that primarily mechanical purification methods and short-term culture are not sufficient to eliminate highly immunogenic cells. In addition, short-term culture is deleterious to the isolate if a significant number of acinar cells is still present after enrichment.

Adult↗