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Biomedical subjects

H Wang

Publications and source records attributed to H Wang.

At least 523 records · Page 29Linked to original sources

Surfactant protein D-coated Klebsiella pneumoniae stimulates cytokine production in mononuclear phagocytes.

Encapsulated Klebsiella pneumoniae strains K21a, K10, and K50, all of which contain dimannose sequences in their capsular polysaccharides that are recognized by the mannose receptor of macrophages, stimulated interleukin secretion and cytokine mRNA expression by human monocyte-derived macrophages. By contrast, the corresponding unencapsulated phase variants and the K2 strain, which lack the dimannose sequence, did not. Coating of unencapsulated phase variants of Klebsiella strains with surfactant protein (SP)-D resulted in marked stimulation of cytokine mRNA accumulation. The induction of cytokine mRNA via the mannose receptor occurred only in monocyte-derived macrophages, whereas that caused by SP-D-coated Klebsiella strains occurred in both macrophages and peripheral-blood monocytes. The results suggested that innate immunity against pulmonary pathogens might be mediated by SP-D, which acts as an opsonin to enhance the interaction of macrophages with unencapsulated phase variants originating from the upper respiratory tract, and by macrophage mannose receptors, which recognize encapsulated variants expressing capsular dimannose residues.

Cytokines↗

[Establishment of ELISA with TT virus(TTV) and its application to patients with hepatitis and healthy people].

OBJECTIVE: To establish the EIA method for detection of antibody to TT virus (anti-TTV) and to investigate the distribution of anti-TTV in patients with hepatitis and in healthy people. METHODS: The ELISA method using the recombinant expressed product of truncated gene of TTV ORF1 as antigen was developed and was applied to detect anti-TTV in patients with hepatitis and in healthy control. The TTV DNA were detected by nested-PCR. RESULTS: The positive rates of anti-TTV antibodies in different populations were as follows: 1.3% in healthy people, 10.5% in patients with hepatitis A, 12.5% in patients with hepatitis B, 8.3% in patients with hepatitis C, 7.7% in patients with hepatitis D, 12.0% in patients with hepatitis E, 6.5% in patients with hepatitis G, 32.3% in patients with hepatitis non-A to G. The positive rate of TTV DNA was significantly related to that of anti-TTV (P < 0.05). CONCLUSION: The anti-TTV antibodies were detected positive in patients with hepatitis and healthy people, however, its positive rate in patients with hepatitis non-A to G was significantly higher than that in other groups. The fact that anti-TTV antibodies coexist with TTV DNA suggests that the antibody to TTV ORF1 antigen may be an infectious marker of TTV similar to that of anti-HCV.

Antibodies, Viral↗

Identification of pelvic lymph nodes with chlorophyllin after injection into the uterine cervix: an experimental and clinical study.

We investigated the value of staining retroperitoneal lymph nodes with chlorophyllin in normal dogs and in women with malignant uterine tumors undergoing lymphadenectomy. In dogs, after 0.3% chlorophyllin (sodium copper chlorophyllin) was injected into the canine uterus, the concentration of dye in the bloodstream was measured with a spectrophotometer and sections of stained retroperitoneal lymph nodes were examined using light and electron microscopy. The highest blood levels were detected at 4 hrs and nearly all of the chlorophyllin was gone from the bloodstream by 18 hrs but was retained in nodal macrophages for at least 4 days. No morphological changes were found in the excised lymph nodes. Twenty-four patients with cervical carcinoma and 20 patients with endometrial carcinoma undergoing radical hysterectomy and lymphadenectomy were divided into a lymphatic coloration group (23 patients) and a non-coloration (control) group (21 patients). In the lymphatic coloration group (0.3% chlorophyllin) was injected into the cervix 5 days before elective lymphadenectomy. There were no complications attributed to injection of the chlorophyllin. The number of dissected lymph nodes in the coloration group were greater than the control group (p<0.01) and the time of operation was shorter (p<0.01). These results suggest that chlorophyllin is safe and facilitates identification of retroperitoneal lymph nodes, allows more complete nodal excision and shortens the time of operation in patients undergoing radical hysterectomy with lymphadenectomy.

Adult↗

In vitro Arabidopsis pollen germination and characterization of the inward potassium currents in Arabidopsis pollen grain protoplasts.

The focus of this study is to investigate the regulatory role of K(+) influx in Arabidopsis pollen germination and pollen tube growth. Using agar-containing media, in vitro methods for Arabidopsis pollen germination have been successfully established for the first time. The pollen germination percentage was nearly 75% and the average pollen tube length reached 135 microm after a 6 h incubation. A decrease in external K(+) concentration from 1 mM to 35 microM resulted in 30% inhibition of pollen germination and 40% inhibition of pollen tube growth. An increase in external K(+) concentration from 1 mM to 30 mM stimulated pollen tube growth but inhibited pollen germination. To study how K(+) influx is associated with pollen germination and tube growth, regulation of the inward K(+) channels in the pollen plasma membrane was investigated by conducting patch-clamp whole-cell recording with pollen protoplasts. K(+) currents were first identified in Arabidopsis pollen protoplasts. The inward K(+) currents were insensitive to changes in cytoplasmic Ca(2+) but were inhibited by a high concentration of external Ca(2+). A decrease of external Ca(2+) concentration from 10 mM (control) to 1 mM had no significant effect on the inward K(+) currents, while an increase of external Ca(2+) concentration from 10 mM to 50 mM inhibited the inward K(+) currents by 46%. Changes in external pH significantly affected the magnitude, conductance, voltage-independent maximal conductance, and activation kinetics of the inward K(+) currents. The physiological importance of potassium influx mediated by the inward K(+)-channels during Arabidopsis pollen germination and tube growth is discussed.

Arabidopsis↗

[High polymorphism at the human P gene locus in Chinese Han and Tibetan populations].

OBJECTIVE: The authors studied two synonymous sites polymorphism (A355A, G780G) and one intervening sequence polymorphism(IVS13-15) of human P gene in 181 and 171 individuals sampled from Chinese Han and Tibetan populations respectively. METHODS: Genomic DNA was extracted from the ACD-blood specimens collected from 352 healthy unrelated individuals by phenol-chloro methods. The amplification of three fragments was performed using PCR technique; the product was digested with restricted enzyme. Agarose electrophoresis was used for typing the PCR product. RESULTS: There were two alleles at each exon variety locus; the allele frequencies in both A355A and G780G were of significant differences between the two populations ( chi(2)=24.54,chi(2)=37.05,P>0.001). No significant difference was found in allele frequency in IVS13-15 locus between the two populations (chi(2)=2.06,P>0.05). And in each of the populations, no significant difference was noted in the genotype frequency between the male and the female. The test for Hardy-Weinberg equilibrium showed that the genotype distributions observed in the two populations were correspondent with the expected. CONCLUSION: There is a remarkable difference in the distribution of allele frequencies at P gene locus between Chinese Han and Tibetan populations, which is related to the variation in skin pigmentation of these two Chinese population.

Adolescent↗

[The expression of HCV RNA polymerase in E.coli and the study of its solubility and antigenicity].

OBJECTIVE: To express HCV RNA polymerase and study its soluble condition and antigenicity. METHODS: We constructed expression vectors pQE-5B-Fl and pQE-5B-C21 and expressed them in E.coli (M15). We analyzed their solubility in different conditions and purified soluble pQE-5B-C21 protein by Ni-NTA column, then detected its antigenicity by ELISA and western blot. RESULTS: We obtained the purified soluble pQE-5B-C21 protein in the induction conditions of 18 degrees C and the protein was proved to be of good antigenicity by ELISA and western blot. CONCLUSIONS: The RNA polymerase of HCV expressed in E.coli has good solubility and antigenicity.

Blotting, Western↗

[Genotypes of transfusion-transmitted virus].

OBJECTIVE: To investigate the genotypes of transfusion-transmitted virus (TTV) in South China. METHODS: One capture probe for all types of TTV from the conserved sequence nt2160 to nt2196 and six varied probes, heterogeneity more than 20% between each other for G1, G2, G3, G4, G5 and G6 type, respectively were synthesized. The primers coming from the conserved sequence of ORF1 were used to amplify TTV DNA from the serums of patients. Microplate sandwich hybridization-ELISA technique was used to detect the genotypes of TTV in 380 hepatitis patients with liver damage. RESULTS: Sixty-one of 380 patients, about 16.0%, were TTV DNA positive, in whom 44 were genotype I (G1), 5 were G2, 10 were mixed infection of G1 and G2, one was G3 and one was G4. Neither G5 nor G6 has been detected. The rate of TTV infection in chronic hepatitis patients was equal to that in acute hepatitis patients. CONCLUSIONS: TTV infection is common in South China. There are four types of TTV. The first genotype was G1, and the second was G2. G5 or G6 has not yet been detected. PCR microplate sandwich hybridization-ELISA has certain value in clinic diagnosis for its specific, sensitive and practicable characters.

DNA Virus Infections↗

[Cloning and expression of maltooligosyltrehalose synthase from Sulfolobus acidocaldarius in Escherichia coli].

The gene of MTSase (maltooligosyltrehalose synthase) from Sulfolobus acidocaldarius ATCC49426 was amplified by PCR. The primers were designed according to the published sequence of homologous gene from Sulfolobus acidocaldarius ATCC33909. This gene was inserted into the plasmid pBV220 and the resultant recombinant plasmid pBV220-GT was transformed to E. coli DH5 alpha. The activity of recombinant enzyme was about 10 u/g(wet cell). In order to improve the expression level of target protein, some nucleotides in the 3' and 5' of the gene were modified to optimize the second structure of mRNA by PCR amplification using the new primers devised according to the biosoftware GOLDKEY2.0. As a result, the activity of recombinant enzyme increase to 19.8 u/g(wet cell). Then, the helping plasmid pUBS520 which carried the gene encoding the tRNA of rare codons AGG and AGA was transformed to the recombinant strain. But it took little effect.

Cloning, Molecular↗

[Studies on mimotopes of hepatitis C virus E1 protein].

OBJECTIVE: To study the B-cell epitope of E1 protein of hepatitis C virus. METHODS: By induction of IPTG, the E.coli M15 strains harboring the pQE30-HCVe118 expressed truncated C-terminal HCV E1 protein (Pte1). The proteins were purified with preparative electrophoreses system, which captured anti-E1 IgG in HCV (+) sera. By applying the antibodies as selective molecular 12 mers random peptide libraries were panned, and positive clones were obtained by ELISA. Amino acid sequences of display peptide were compared with that of HCV E1 protein. RESULTS: The purified HCV E1 proteins could react specifically with partly anti-HCV sera by ELISA. Among 10 phage display peptides, 6, 6, 2 were the most homologous to HCV E1 protein at position 320-336aa, 251-263aa, 225-248, respectively. CONCLUSIONS: There exist multiple B-cell epitopes in HCV E1 protein. At least one preponderant epitope is mapped at residues 320-336 of HCV E1 protein.

Amino Acid Motifs↗

[Simulation study of electrode configurations for cardiac stroke volume calculation by 3-dimensional finite element method].

This is a research in Kubicek electrode configurations for cardiac stroke volume calculation from the angle of Kubicek model simulation by 3-dimensional finite element method. In the process of computer simulation, we have studied, respectively, the effect of the width, thickness, and material property (specific resistance) of potential electrode band on cardiac stroke volume calculation in order to guide the choice of electrode configurations in the clinic. The simulation results showed that in Kubicek electrode configurations, its all properties of the potential electrode band affected the impedance results for cardiac stroke volume calculation, one main factor of which is the choice of the material property. Therefore, we have to attend to the choice of Kubicek electrode configurations while calculating SV by Kubicek method in order to improve the accuracy of SV calculation.

Electrodes↗

[Design and implementation of mini PACS in the hospital].

This paper introduces the principle and classification of picture archiving and communication system (PACS) and some pertinent technologies, including the capture methods of pictures data, images storage equipment and the choice of medium, and the network of images transmission. Also presented is an analysis of the merit and demerit of different kinds of PACS, and in the end we put forward a PACS design method and network framework based on practical implication in the Jinwei project experimental hospital.

Computer Communication Networks↗

Effects of ouabain and digoxin on gene expression of sodium pump alpha-subunit isoforms in rat myocardium.

OBJECTIVE: To compare the effects of ouabain and digoxin on the gene expression of sodium pump alpha-subunit isoforms in the myocardium of rats. METHODS: Normal Sprague-Dawley (SD) rats were injected with ouabain (20 micrograms.kg-1.d-1, i.p.), digoxin (32 micrograms.kg-1.d-1, i.p.) and normal saline (NS) once a day, respectively, and indirect systolic blood pressure was recorded once a week. Six weeks later, all of the rats were killed, and sodium pump alpha 1-, alpha 2-, and alpha 3-subunit mRNA levels in the myocardium were detected with the reverse transcription polymerase chain reaction (RT-PCR) method. RESULTS: The systolic blood pressure of the rats infused with ouabain increased significantly at the end of week 6 (132.6 +/- 9.0 mm Hg vs 115.7 +/- 8.2 mm Hg, P < 0.01), while no difference in blood pressure was found between the digoxin group and the NS group. The expression of sodium pump alpha-subunit isoforms in the ventricular myocardium was regulated by either ouabain or digoxin. Both ouabain and digoxin stimulated expression of the alpha 3-isoform, whereas alpha 2 was unchanged in those two groups. alpha 1-isoform expression decreased in the ouabain group and was unchanged in the digoxin group. CONCLUSIONS: These results suggest that both ouabain and digoxin could regulate sodium pump alpha-subunit isoform expression, which might be related to the physiological roles of endogenous ouabain and might be responsible for the difference in the pharmacological and toxicological effects of ouabain and digoxin, including their effects on blood pressure.

Animals↗

The cAMP-mediated protein kinase signal transduction pathway is involved in the pyrogenic effect of CRH in rats.

OBJECTIVE: To determine whether the cyclic adenosine monophosphate (cAMP) mediated protein kinase signal transduction pathway is involved in the pyrogenic action of corticotropin releasing hormone (CRH) in rats. METHODS: Corticotropin releasing hormone, 2',3'-dideoxyadenosine (DDA) and adenosine-3',5'-(cyclic) monophosphorothionate, Rp-Isomer (Rp-cAMPS), were administered intracerebroventricularly (i.c.v.). The colonic temperature was measured using a thermistor, and the content of cAMP in the hypothalamus was determined by radioimmunoassay. Hypothalamic incubation was used to assess the effects of CRH on the content of cAMP in the hypothalamus in vitro. RESULTS: Microinjection (i.c.v.) of CRH (2.5 micrograms, 5.0 micrograms and 10 micrograms) caused increases in colonic temperature and the hypothalamus cAMP level in conscious rats. CRH increased hypothalamus cAMP level in vitro. The pyrogenic effects of CRH were abolished or markedly inhibited by prior injection (i.c.v.) of an adenylate cyclase inhibitor, DDA (30 micrograms), or an inhibitor of cAMP-dependent protein kinase, Rp-cAMPS (15 micrograms). CONCLUSION: cAMP mediates the pyrogenic action of centrally administered of CRH in rats, and protein kinase A may play an important role in the central CRH-induced fever. The cAMP-dependent protein kinase signal transduction pathway may be involved in the central mechanisms of the pyrogenic action of CRH in rats.

Animals↗

Short-term study of allogeneic retinal pigment epithelium transplants onto debrided Bruch's membrane.

PURPOSE: To investigate the survival and behavior of retinal pigment epithelium (RPE) microaggregates transplanted onto hydraulically debrided Bruch's membrane and to compare results of using three different vehicles for cell delivery. METHODS: RPE microaggregates obtained from male cats were transplanted onto the tapetal area of female cats after native RPE was debrided. For the control, one of three vehicles was introduced into the debridements. Each transplant or control specimen was analyzed histologically and immunohistochemically. Transplanted male RPE cells were identified by in situ labeling of the cat Y chromosome. RESULTS: Histologically, significant numbers of condensed, darkly stained RPE nuclei were observed in all transplants compared with few TUNEL-positive RPE cells. Cellular retinaldehyde-binding protein was present up to day 7 in all RPE cells in transplants. In both transplant and control specimens, the antibody against the Ki-67 nuclear antigen labeled some RPE cells at day 3. TUNEL-positive outer nuclear layer nuclei were most frequently observed at day 1, but were much less frequent at 7 days in both transplant and control specimens. CONCLUSIONS: Transplanted RPE appeared to retain at least some markers of differentiation up to 7 days after surgery. Some proliferation of transplanted RPE cells was also seen. Apoptotic cell death of transplanted RPE, as judged by TUNEL staining was observed rarely. RPE transplants imposed no adverse effect on the overlying retina. RPE survival appeared to be similar with each of the three vehicles for cell delivery.

Animals↗

[Construction of plant expression vectors harboring a peptide antibiotic-apidaecin gene and resistance analysis of the transgenic tobacco].

Two plant expression vectors(pBinPRHbI and pBinPRSIHbI) were constructed: Firstly, apidaecin gene were fused to the signal peptide coding sequencing of a PR-protein, and cloned into a binary vector pBin438 to form pBinPRHbI. Then, the cassette consisting of 35S promoter, PR signal peptide coding sequencing and apidaecin gene was cut off from pBinPRHbI and inserted into another plant expression vector pBinPRSI to produce a bivalent plant expression vector pBinPRSIHbI. pBinPRSI was constructed previously in our lab and contained PR signal peptide and Shiva-I fusion gene under control of 35S promoter. The three plant expression vectors were introduced into tobacco by Agrobacterium-mediated transformation. The positive rate of PCR was 95% in all putative transgenic plants. Results from Southern blot indicated that foreign genes were integrated into tobacco genome and RT-PCR analysis proved that the foreign gene was transcribed in transgenic tobacco. The transgenic tobacco showed higher resistance to P. syringae pv tabaci, the causal agent of tobacco wild fire disease, than their original cultivars. From the disease index, the transgenic plants carrying apidaecin and Shiva-I genes had highest resistance among three kinds of transgenic plants, and the plants carrying Shiva-I gene alone had lowest resistance.

Anti-Infective Agents↗

Antisense anti-MDM2 oligonucleotides as a novel therapeutic approach to human breast cancer: in vitro and in vivo activities and mechanisms.

The mouse double minute 2 (MDM2) oncogene has been suggested as a target for cancer therapy. It is amplified or overexpressed in many human cancers, including breast cancer, and MDM2 levels are associated with poor prognosis of several human cancers, including breast cancer, ovarian cancer, osteosarcoma, and lymphoma. In the present study, we investigated the functions of MDM2 oncogene in the growth of breast cancer and the potential value of MDM2 as a drug target for cancer therapy by inhibiting MDM2 expression with a specific antisense antihuman-MDM2 oligonucleotide (oligo). The selected antisense mixed-backbone oligo was evaluated for its in vitro and in vivo antitumor activity in human breast cancer models: MCF-7 cell line containing wild-type p53 and MDA-MB-468 cell line containing mutant p53. In MCF-7 cells, p53 and p21 levels were elevated, resulting from specific inhibition of MDM2 expression by the antisense oligo (AS). In MDA-MB-468 cells, after inhibition of MDM2 expression, p21 levels were elevated, although p53 levels remained unchanged. After i.p. administration of the antisense anti-MDM2 oligo, in vivo antitumor activity occurred in a dose-dependent manner in nude mice bearing MCF-7 or MDA-MB-468 xenografts. In both models, in vivo synergistically or additive therapeutic effects of MDM2 inhibition and the clinically used cancer chemotherapeutic agents irinotecan, 5-fluorouracil, and paclitaxel (Taxol) were observed. These results suggest that MDM2 have a role in tumor growth through both p53-dependent and p53-independent mechanisms. We speculate that MDM2 inhibitors, such as ASs, have a broad spectrum of antitumor activities in human breast cancers, regardless of p53 status. This study should provide a basis for future development of anti-MDM2 ASs as cancer therapeutic agents used alone or in combination with conventional chemotherapeutics.

Animals↗

[Studies on chemical constituents of mycelium of fungus Cephalosporium sp. AL031(I)].

Five compounds were isolated from the mycelium of the fungus Cephalosporium sp. AL031 whose metabolites have been proven to possess antifungal and antibacterial activities. Based on the spectral data and elemental analysis, they were identified as ergosterol(I), 2,4,6-octatrienoic acid(II), succinic acid(III), uracil(IV), and D-mannitol(V). All of these compounds were obtained from the culture of this fungus for the first time.

Acremonium↗