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Biomedical subjects

J Zhou

Publications and source records attributed to J Zhou.

At least 577 records · Page 32Linked to original sources

[The role of nitric oxide in hyperdynamic circulation in portal hypertensive rats].

OBJECTIVE: To evaluate the role of nitric oxide (NO) in hyper-hemodynamics in portal hypertension, we investigated the effects of nitric oxide synthase inhibitor L-NMMA with or without L-arginine, the substrate for enzyme NO synthase on the systemic and splanchnic hemodynamics in portal hypertensive rats. METHOD: Ninety-seven male Sprague-Dawley rats were divided into four groups: intrahepatic portal hypertension (IHPH, n = 23) by injection of CCl4, prehepatic portal hypertension (PHPH, n = 26) by stenosis of the portal vein, end-to-side portacaval shunt (PCS, n = 23), and sham-operated controls (SO, n = 25). Animals of each group were divided into three subgroups: systemic administration of L-NMMA (50 mg/kg BW, 1 ml), L-NMMA (50 mg/kg BW, 0.5 ml) plus L-arginine (30 mg/kg BW, 0.5 ml), and normal saline solution (1 ml). The radioactive microsphere method was used for hemodynamic study. RESULT: There was the characteristic of hyperdynamic circulatory state including increased cardiac output and splanchnic blood flow, decreased mean arterial blood pressure, total peripheral vascular resistance and splanchnic vascular resistance in IHPH, PHPH and PCS rats. The hyperdynamic circulatory state in IHPH, PHPH and PCS rats could be effectively reversed by L-NMMA to the resting values of hemodynamics in SO rats, but not the levels after administration of L-NMMA in SO rats, indicating that there is excessive production of NO in portal hypertensive and PCS rats. Administration of L-arginine counterbalanced the effects of L-NMMA on hyperdynamic circulatory state in portal hypertensive and PCS rats. CONCLUSION: NO is an important mediator of hyperdynamic circulation in portal hypertension.

Animals↗

[Retrograde partial hepatectomy: report of 11 patients].

OBJECTIVE: To study a new approach to hepatectomy for liver tumors resected difficulty by typical method. METHOD: The operative order is retrograde as compared with typical hepatectomy, namely, transection of the liver is performed first, isolating adhesions between the tumor and the diaphragm or partial invaded phrenectomy is performed second, and then after cutting corresponding ligaments, liver tumor is removed. If the surrounding organs were invaded or adhered by tumor too tightly to be separated. They were resected with the tumor. RESULT: The approach was used in 11 patients with malignant tumors of liver from June 1994 to January 1996. Operation time was 192 min (150 - 250 min) and estimated blood loss during operation was 1460 ml (800 - 4, 200 ml). No operative mortality was found and all of the patients recovered uneventfully. CONCLUSION: The retrograde partial hepatectomy is a good method for resection of liver tumors resected difficulty by typical hepatectomy.

Adult↗

[A study on lens epithelial cell ultrastructure in rhesus monkey cataract].

OBJECTIVE: To investigate the ultrastructure of lens epithelial cells of normal eyes and cataractous eyes in rhesus monkeys. METHODS: Anterior capsules were taken from 4 normal lenses and 4 cataractous lenses of rhesus monkey eyes, and the samples were observed under transmission electron microscope. RESULTS: The ultrastructure of the cataractous lens epithelial cells changed obviously. Mitochondria with disappearance of crista were swollen, and vacuolar. Hollow cell with a single large vacuole in the cytoplasm could be seen. The cytoplasm presented dissolution. Cells were swollen and damaged, even cell membrane disruption could be seen. Karyopyknosis, nuclear deformity, disappearance of perinuclear cistern and nuclear pores could be found. The heterochromatin was concentrated and aggregated at the nuclear periphery. CONCLUSIONS: The ultrastructural changes of the lens epithelial cells in monkey cataractous lens are similar to that in human cataractous lens. Rhesus monkey cataract may be a suitable experimental model to investigate the formation of human cataract.

Animals↗

[Application of ultrasound microscope for the reattachment surgery of detachment of ciliary body].

OBJECTIVE: To evaluate the advantages of ultrasound biomicroscope (UBM) used for the reattachment surgery of traumatic ciliary body detachment. METHODS: 33 cases diagnosed by UBM as ciliary body detachment were studied. The surgical results of these cases were analyzed and compared with that of 83 cases which were simply diagnosed by a gonioscope. RESULTS: 31 of the 33 cases regained the normal intraocular pressure (IOP) after once of operation (94%), and all cases achieved successful results after twice of operation (100%). Of the 83 cases examined only by gonioscopy, the rates of reaching normal IOP after the 1(st), 2(nd) and 3(rd) operations were 82%, 94% and 96% respectively. CONCLUSIONS: These results suggest that the UBM be useful for the tiny and narrow detachment of ciliary body which can not be seen by a gonioscope. UBM has important clinical value in diagnosis, guide to the range of suture and follow-up observation. With the use of UBM, the frequency of operation necessary for surgical success and blindness in action in the operation will be reduced, and the successful rate of once of operation will be increased significantly.

Adolescent↗

[Sulfation of polysaccharides isolated from Indocalamus tesselatus and their anticytopathic effect on human immunodeficiency virus type I].

For preparation of sulfated polysaccharides isolated from bamboo leaves (Indocalamus tesselatus) with chlorosulfonic acid-pyridine, the experimental conditions as the molar ratio of reactants and temperature and time of reaction were discussed. A homogeneous sulfation method with good yield of more than 80% was obtained and the sufur content was found to be 21.9%. Conformation change in sulfated polysaccharide with changes of intrinsic viscosity, polarimetery and CD spectra were found to be related to the negative charge. The polysaccharide sulfates were shown to have better activity to inhibit cytopathic effect of human immunodeficiency virus(HIV) infection than native polysaccharides. The effective concentration was 10 micrograms.ml-1 while the cytotoxic concentration was 2.5 mg.ml-1.

Antiviral Agents↗

[Preparation and characterization of monoclonal antibodies against Clonorchis sinensis].

AIM: To develop and identify monoclonal antibody (McAb) against adult Clonorchis sinensis. METHODS: BALB/c mice were immunized with soluble antigen of adult Clonorchis sinensis, the spleen cells of immunized mice were fused with myeloma cell line SP2/0. Then the hybridoma cell lines secreting high titer of McAbs were screened. The Ig subtypes and titer of the McAbs were assayed with ELISA. The antigens recognized by the McAbs were examined with IFAT and identified by immunoblot analysis. The cross reactions between the McAbs and antigens of Schistosoma japonicum, Paragonimus westermani and Cysticercus cellulosae were detected. RESULTS: 5 hybridoma cell lines secreting high titer of McAbs against adult Clonorchis sinensis were established. All five clones of McAbs belonged to IgM subtype. The antigens recognized by the McAbs were located at the intestinal tract wall of adult Clonorchis sinensis. The McAbs had no cross reactions with antigens of Schistosoma japonicum, Paragonimus and Cysticercus cellulosae. CONCLUSION: 5 hybridoma cell lines against adult Clonorchis sinensis secreting high titer of McAbs with high specificity were established.

Animals↗

Immunoscreening of Schistosoma japonicum egg cDNA library.

AIM: To screen and identify genes encoding vaccine candidates and molecules with diagnostic potential from S. japonicum (S. j) egg cDNA library. METHODS: S. j egg cDNA library was screened by sera from SEA-immunized rabbits. Positive clones were identified after three rounds of screening and were excised automatically in vivo with helper phage. Products of PCR were used to detect insert sizes by agarose gel electrophoresis. DNA samples were obtained by purification using mini-prep kit. Automatic sequencing was carried out and the sequence data were analysed using GCG software. RESULTS: Twelve clones were identified as positive and the inserts were around 1.2 kb in size. Compared with DNA sequences in GenBank, eight clones were homologous to a 70 kDa S. j heatshock protein(S. j HsP70). C18 exhibited 97.8% homology with S. j23. C22 and C23 shared 82.4% and 89.5% homology with S. j calreticulin, respectively. C12 was homologous to S. mansoni(S. m.) immunophilin. CONCLUSION: This paper first reported the results of immunoscreening S. j egg cDNA library by anti-SEA sera. Twelve positive clones were found to be related to protective immunity and/or immunodiagnosis of schistosomiasis.

Animals↗

[DNA cloning of Ad7 vaccine strain and sequence analysis of its Sma I H fragment].

Human adenoviruses have been a hot topic of study because of their potential utility for gene therapy and development of live viral vectored vaccines. Adenovirus types 4 and 7 have been given orally as vaccines for prevention of acute respiratory infections caused by these serotypes of adenovirus in U.S. army recruits for more than 30 years. Therefore, the advantage of using Ad4 and Ad7 as vectors in the development of oral live recombinant vaccine is obvious. For constructing Ad7 vector, 10.8-100 mu DNA of Ad7 vaccine strain has been cloned. The Sma I H fragment was sequenced using Ad7 DNA as a template and then the fragment was cloned by PCR. For verifying the sequence the fragment was sequenced again. The strategy we used to clone terminal end of adenovirus is reliable and practicable.

Adenovirus E3 Proteins↗

[Sequence analysis of Ad7 vaccine strain 87 mu-97.4 mu fragment].

Adenovirus 7 vaccine strain(Ad7v) possesses potentiality to be used as a vector for producing recombinant vaccine. In order to construct Ad7 vector, the nucleotide sequence of a 3698 bp fragment(87 to 97.4 mu) of the virus DNA was analysed. The fragment encodes three ORFs namely, one ORF of early region 4(E4), one ORF of early region 3 and one ORF of fiber protein. The predicted amino acids of three ORFs(ORF 8.1 kD, 42.8 kD and 10.3 kD) share 33%, 43% and 44% homology with Ad2 E4 corresponding regions (ORF 3, ORF 4 and ORF 6/7) respectively. A comparison of the nucleotide and amino acid sequences of those fiber ORF demonstrated a higher homology between Ad7v and prototype strains of Ad7v than those between Ad7v and Ad7h. The Ad7v E3 15.4 kD protein shares 86%, 88%, 90% and 98.5% amino acid homology with the equivalent region of ad7h E3 15.3 kD protein, Ad4 E3 region 15 kD protein, Ad3 E3 15.3 kD and Ad7p E3 15.3 kD protein protein respectively, but only 68% with Ad2 E3 14.7 kD protein.

Adenoviridae↗

[Cloning, sequencing and expression of hepatitis E virus structural gene in E. coli and application of the recombinant products for diagnosis].

The prokaryotic expression vector PEGX-3X was used to express hepatitis E virus (HEV) open reading frame 2(ORF2 402-660). The recombinant protein was soluble in PBS buffer and was purified by the glutathione Sepharose 4B affinity column. When the purified recombinant protein was applied to detect HEV antibodies in sera of clinical patients, the results were quite consistent with the HEV diagnostic kit from Diagnostic Biotechnology Ltd(DBL), Singapore. The recombinant protein was more reactive with HEV antibodies than DBL reagent.

Antigens, Viral↗

[luxAB genes as marker for detecting Rhizobium fredii HN01 nodulation functions].

A suicide plasmid pHNC3 which contains Tn5-luxAB was transferred into Rhizobium fredii HN01 by the help of pRK2013. Then Tn5-luxAB inserted on the genome of HN01 and gave luminescence activity. The luminescence colonies were picked up and the Eckhardt gel was performed for plasmids profile detection. The location of Tn5-luxAB on the genome was determined using the luxAB as probe. The colonies which were marked by Tn5-luxAB on the chromosome and different plasmids of HN01 were chosen for pot experiment, and a chromosome labelled strain HN01LC02 was detected by soil pot experiment. The detections included the nodulation occupancy and the luminescent nodules distribution on the root system formed by the luxAB-marked rhizobia.

Chromosomes, Bacterial↗

Expression of heterogeneous nuclear ribonucleoprotein A2/B1 in bronchial epithelium of chronic smokers.

The monoclonal antibody 703D4, which binds heterogeneous nuclear ribonucleoprotein A2/B1 (hnRNP A2/B1), has been reported to detect lung cancer more than a year earlier than routine chest X-ray or cytomorphology. To explore the biological basis of this detection, we studied the expression of this antigen in the central airways of smokers with evidence of bronchial metaplasia using specimens from a previously reported, randomized retinoid chemoprevention trial. By analyzing 1078 available biopsy specimens from 147 individuals at baseline and 68 individuals who completed the intervention, we frequently detected overexpression of hnRNP A2/B1 in normal and abnormal bronchial epithelium (i.e., in 41% of normal and 37% of squamous metaplasia samples). There was no correlation between hnRNP A2/B1 overexpression and the different histological changes. In cases with hnRNP A2/B1 overexpression, immunoreactivity was homogeneously expressed in all biopsied sites. For the 68 cases with serial biopsies, there was no significant modulation of hnRNP expression by retinoid intervention or smoking status. With lung cancer cell lines, 0.5-4 microM concentrations of 13-cis-retinoic acid reduced hnRNP A2/B1 overexpression by immunocytochemistry. We conclude that hnRNP A2/B1 overexpression is frequently found in central airways of chronic smokers, consistent with the pattern of expression that we reported previously in airways surrounding resected primary lung cancers. Oral 13-cis-retinoic acid at a dose of 1 mg/kg has no demonstrable effects on modulating hnRNP A2/B1 expression in proximal bronchial epithelium.

Biomarkers, Tumor↗

Blood volume redistribution on fluid treatments after burn-blast combined injury, especially with respect to the regulation role of microcirculation due to the Fåhraeus effect.

Considering the Fåhraeus effect and the blood volume regulation role of the microcirculation, we used a new method for calculating the blood volume change in microcirculation, macrocirculation as well as the whole circulation (delta Vmic, delta Vmac, delta Vb), to compare fluid therapy effects of hypertonic saline dextran solution (HSD) and lactic Ringer's solution at the early stage after burn-blast combined injury (BBI). The measurement of plasma viscosity was used in this calculation. The results showed that, with calculation of the blood volume change in microcirculation and macrocirculation, this method could more exactly and distinctly display the change and distribution of blood volume during the therapy. It confirms that HSD treatment can increase blood pressure and attenuate tissue edema, by significantly increasing total blood volume, recouping delta Vmac. The study suggests that a desirable and practical clinical method for blood volume change can be developed based on the present study.

Animals↗

Failure to change exploration or saccharin preference in rats exposed to chronic mild stress.

Chronic mild stress (CMS) exposes animals to unpredictable stressors. Reduced consumption of sucrose or saccharin solutions by CMS rats has been used as a putative measure of anhedonia, typical of depression. Our objective was to determine whether saccharin consumption and preference and suppression of exploratory and rearing behaviors in the open field were reliable indicators of CMS-induced behavioral depression. In Experiment 1, male Wistar rats subjected to 6 weeks of CMS consumed significantly less food and gained less weight than controls. CMS did not effect saccharin intake, or preference, measured in a two-bottle test with water. CMS rats exposed to a novel open field showed increased exploration and rearing. In a second test, performed immediately after a novel stress of restraint, there were no differences in exploratory or rearing behavior of CMS and control rats. In Experiment 2, CMS was reduced to 3 weeks and rats were single or group housed in their home cages. Open field activity of CMS rats was similar to that in Experiment 1. Saccharin preference of CMS rats was significantly suppressed when tested after 24 hours of water deprivation, but was not different from controls after 5 hours of water deprivation. In the final experiment Sprague Dawley rats behaved the same as Wistar rats in the CMS paradigm. Therefore, the CMS protocol used in these experiments did not induce behaviors indicative of depression but did cause a mild anorexia and weight loss. Saccharin intake of CMS rats was dependent upon their dehydration state and could not be attributed to stress-induced anhedonia.

Animals↗

The degree of nonlinearity and anisotropy of blood vessel elasticity.

Blood vessel elasticity is important to physiology and clinical problems involving surgery, angioplasty, tissue remodeling, and tissue engineering. Nonlinearity in blood vessel elasticity in vivo is important to the formation of solitons in arterial pulse waves. It is well known that the stress-strain relationship of the blood vessel is nonlinear in general, but a controversy exists on how nonlinear it is in the physiological range. Another controversy is whether the vessel wall is biaxially isotropic. New data on canine aorta were obtained from a biaxial testing machine over a large range of finite strains referred to the zero-stress state. A new pseudo strain energy function is used to examine these questions critically. The stress-strain relationship derived from this function represents the sum of a linear stress-strain relationship and a definitely nonlinear relationship. This relationship fits the experimental data very well. With this strain energy function, we can define a parameter called the degree of nonlinearity, which represents the fraction of the nonlinear strain energy in the total strain energy per unit volume. We found that for the canine aorta, the degree of nonlinearity varies from 5% to 30%, depending on the magnitude of the strains in the physiological range. In the case of canine pulmonary artery in the arch region, Debes and Fung [Debes, J. C. & Fung, Y. C.(1995) Am. J. Physiol. 269, H433-H442] have shown that the linear regime of the stress-strain relationship extends from the zero-stress state to the homeostatic state and beyond. Both vessels, however, are anisotropic in both the linear and nonlinear regimes.

Animals↗

Proper spacing between heptad repeat B and the transmembrane domain boundary of the paramyxovirus SV5 F protein is critical for biological activity.

The paramyxovirus, simian virus 5, fusion (F) protein contains seven amino acids between heptad repeat B (a domain required for a biologically active fusion protein) and the presumptive boundary of the transmembrane (TM) domain. The role of the seven membrane proximal residues in stability and fusion promotion was examined by construction of a series of insertion, substitution, and deletion mutants, as manipulation of this region to enable proteolytic cleavage would facilitate production of a soluble F protein. The majority of the mutant F proteins both oligomerized and had kinetics of intracellular transport similar to those of wild-type (wt) F protein. All mutant F proteins were expressed at the cell surface at or near the same level as the wt F protein. However, by using both a qualitative lipid mixing assay and a quantitative content mixing assay for membrane fusion, it was found that mutant F proteins containing insertions in the region between heptad repeat B and the TM domain were unable to induce fusion, whereas the mutant F proteins containing substitutions in this region, together with three of the four mutants with deletions in this region, could induce fusion. Four of the F protein mutants contained a Factor Xa cleavage site, IEGR; however, Factor Xa treatment of cell surfaces released either none or only very small amounts (< 1% of total protein) of the soluble heterodimer F1 + F2. As an alternative method of generating soluble F protein, a glycosyl phosphatidylinositol (GPI) anchor was added to the F protein at three membrane-proximal positions. The highest level of surface expression was observed when the final molecule did not contain a significant insertion of amino acids into the membrane proximal region. Two F-GPI mutants reached the surface at approximately 20% of the levels seen with the wt F protein, and approximately 25% of the cell surface population of these mutants could be cleaved with phosphatidylinositol phospholipase C (PI-PLC) to yield soluble F protein. However, all the F-GPI mutants oligomerized aberrantly and failed to promote fusion. Taken together, these data indicate that the spacing of the region immediately adjacent to the presumptive boundary of the TM domain is extremely important for the fusogenic activity of the SV5 F protein.

Animals↗

Participation of ADP dissociation in the rate-determining step in cAMP-dependent protein kinase.

Pre-steady-state kinetic analyses of the catalytic subunit of cAMP-dependent protein kinase showed that the rate constant for phosphoryl transfer is fast and either the release of one or both of the products or a conformational change controls turnover [Grant, B., & Adams, J. A. (1996) Biochemistry 35, 2022-2029]. To determine which step or steps control turnover in the wild-type enzyme, we used a catalytic trapping technique to measure directly the dissociation rate constant for ADP. The phosphorylation of two peptide substrates, LRRASLG and GRTGRRNSI, was monitored using a rapid quench flow technique under conditions where saturating concentrations of ADP were preequilibrated with the enzyme before excess ATP and one of the substrates were added to trap the free enzyme and to start the phosphorylation reaction. Under ADP preequilibration conditions, no 'burst' phase was observed, and although the rate of linear, steady-state turnover was unaffected, the net production of phosphopeptide lagged behind the non-preequilibrated control. This phenomenon occurs due to the slow release of the product, and kinetic modeling suggests that this effect can be explained if the dissociation rate constant for ADP is 24 s-1 and solely limits turnover (kcat = 23 s-1) for the phosphorylation of LRRASLG. Using GRTGRRNSI, the dissociation rate constant for ADP is 35 s-1 and limits turnover (kcat = 29 s-1) if the reaction is initiated by the addition of enzyme. Under preequilibration conditions with either ATP or GRTGRRNSI, turnover is approximately 50% lower, suggesting that ADP release may partially control this parameter. This preequilibration effect can be explained by slowly interconverting enzyme forms with specific peptide-induced turnover properties. These studies indicate that ADP release is an essential rate-limiting component for turnover but also suggests that other factors contribute subtly when the structure of the substrate is altered.

Adenosine Diphosphate↗