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Biomedical subjects

M Shen

Publications and source records attributed to M Shen.

At least 91 records · Page 5Linked to original sources

Structural and phylogenetic analyses of RGD-CAP/beta ig-h3, a fasciclin-like adhesion protein expressed in chick chondrocytes.

A cDNA for RGD-CAP/beta ig-h3 was cloned from a chick embryo chondrocyte cDNA library. The deduced amino acid sequence showed that the chick RGD-CAP/beta ig-h3 is 76-77% identical with human, mouse and pig forms of the protein, and 43% identical with human and mouse osteoblast specific factor 2 (OSF2). RGD-CAP/beta ig-h3 contained four internal repeat domains and two highly conserved sequences (H1 and H2) in each repeat. Chick RGD-CAP/beta ig-h3, as well as the mammalian RGD-CAP/beta ig-h3, contained an RGD sequence, which may serve as a recognition sequence for integrins, in the fourth repeat. Database searches revealed that the H1 and H2 sequences are conserved in some secreted or membrane proteins of several species including mammals, insects, sea urchins, plants, yeast and bacteria. Phylogenetic analysis showed that a portion of the common ancestor gene for RGD-CAP/beta ig-h3 and OSF2 was duplicated to form four repeat domains before the separation of the genes followed by the divergence of vertebrate species.

Amino Acid Sequence↗

The cannabinoid agonist Win55,212-2 inhibits calcium channels by receptor-mediated and direct pathways in cultured rat hippocampal neurons.

The effects of the cannabinoid receptor agonist Win55,212 on Ca2+ channels were studied in rat hippocampal neurons grown in primary culture. Win55,212-2 inhibited whole-cell Ba2+ currents through Ca2+ channels by both CB1 receptor-mediated and direct mechanisms. The concentration dependent inhibition of the current showed two clear phases, a high-affinity receptor-mediated phase (IC50=14+/-2 nM) that was stereoselective and sensitive to a CB1 receptor antagonist, 300 nM SR141716, and a non-saturating phase that was neither stereoselective nor inhibited by SR141716. These concentration-dependent effects were paralleled by Win55212-induced inhibition of glutamatergic synaptic transmission. Win55,212-2 (100 nM) inhibited both omega-agatoxin IVA- and omega-conotoxin GVIA-sensitive currents. Thus, activation of cannabinoid receptors inhibits N- and P/Q-type Ca2+ channels. Activation of cannabinoid receptors inhibited only a fraction of the whole-cell Ca2+ channel current (17+/-2%) even though more than half of the whole-cell Ba2+ current was carried by N- and P/Q-type Ca2+ channels. Concentrations of agonist greater than 1 microM inhibited Ca2+ channels directly.

Analgesics↗

Does glutaraldehyde induce calcification of bioprosthetic tissues?

BACKGROUND: Glutaraldehyde has been said to be responsible in part for the calcification of glutaraldehyde-treated tissues after implantation in animals or humans. We investigated whether the origin of the tissue, autologous or heterologous, could have a more prominent role in the process of calcification. METHODS: Three-month-old sheep received sheep pericardial samples (n = 133) and human pericardial samples (n = 123) implanted subcutaneously. Samples were treated with 0.6% glutaraldehyde for 5, 10, or 20 minutes or 7 days and then rinsed thoroughly before implantation. Samples were then retrieved after 3 months. Calcium content was assessed by spectrophometry. RESULTS: The results show a low calcium content in the autologous group (mean 1.14+/-2.07) and a high calcium content in the heterologous group (mean 38.97+/-26). These results were the same regardless of the duration of the treatment. CONCLUSIONS: Glutaraldehyde treatment (0.6%) does not play a significant role in the calcification of glutaraldehyde-treated tissue regardless of the origin, autologous or heterologous, of the tissue. Glutaraldehyde-treated autologous tissues are associated with an incidence of calcification lower than heterologous tissues.

Analysis of Variance↗

Heat treatment mitigates calcification of valvular bioprostheses.

BACKGROUND: Several techniques have been proposed to mitigate calcification of glutaraldehyde-preserved bioprostheses. None has been fully satisfactory. Knowing that heat induces significant molecular changes, we investigated the potential benefit of the heat treatment of glutaraldehyde-fixed tissues. METHODS: Samples of porcine valves and bovine pericardium treated in 0.625% glutaraldehyde were submitted to temperatures from 37 degrees to 70 degrees C for 2 to 12 weeks and then implanted subcutaneously in newborn rats for 3 months. In a second protocol, nine heat-treated porcine valve bioprostheses and seven control porcine valve bioprostheses were implanted in the mitral position in sheep for 20 weeks. RESULTS: Spectrophotometry and x-ray analysis comparing heat-treated versus non-heat-treated tissues showed the following: porcine valve: 6.7+/-2.3 microg Ca/mg (n = 8) versus 239.9+/-2.9 /g Ca/mg (n = 8); bovine pericardium: 19.5+/-8 microg Ca/mg (n = 6) versus 108+/-10.3 microg Ca/mg (n = 8); and porcine valve bioprostheses: 0 versus ++. CONCLUSIONS: Heat treatment of glutaraldehyde-treated bioprostheses significantly mitigated calcification in both subcutaneous and mitral position.

Animals↗

Infrared spectroscopy in the evaluation of the process of calcification of valvular bioprostheses.

BACKGROUND: The methods currently used to analyze the process of calcification of bioprostheses give only global information on calcium deposition. We investigated the potential advantage of infrared spectroscopy, which makes it possible to analyze the various components of the calcification process, ie, lipids, proteins, and calcium deposits. METHODS: Sixty porcine aortic leaflets were fixed in 0.6% glutaraldehyde and then subsequently implanted in 10-day-old Wistar rats. The valve leaflets were removed 2, 7, 14, 21, 35, and 56 days after implantation. RESULTS: Before implantation infrared spectroscopic analysis revealed the presence of proteins only. On day 2 after implantation, all valves showed minor lipid deposits. On day 7, amorphous calcium phosphate was detected. Between days 7 and 14, crystalline forms of calcium phosphate appeared and amorphous calcium phosphate progressively changed into carbapatite over the 56-day period. CONCLUSIONS: Infrared spectroscopy yields valuable additional information on the nature and kinetics of the various components of glutaraldehyde-treated tissues after implantation. It may prove to be important in the evaluation of new techniques of calcium mitigation.

Animals↗

Cannabinoid receptor agonists protect cultured rat hippocampal neurons from excitotoxicity.

Cannabinoid receptor agonists act presynaptically to inhibit the release of glutamate. Because other drugs with this action are known to reduce excitotoxicity, we tested several cannabimimetics in a model of synaptically mediated neuronal death. Reduction of the extracellular Mg2+ concentration to 0.1 mM evoked a repetitive pattern of intracellular Ca2+ concentration ([Ca2+]i) spiking that, when maintained for 24 hr, resulted in significant neuronal death. The [Ca2+]i spiking and cell death in this model result from excessive activation of N-methyl-D-aspartate receptors, as indicated by the inhibition of both [Ca2+]i spiking and neuronal death by the N-methyl-D-aspartate receptor antagonist CGS19755 (10 microM). The cannabimimetic drug Win55212-2 (100 nM) completely blocked [Ca2+]i spiking and prevented neuronal death induced by low extracellular Mg2+ concentrations. These effects on [Ca2+]i spiking and viability were stereoselective and were prevented by the CB1 receptor antagonist SR141716 (100 nM). The partial agonist CP55940 (100 nM) also afforded significant protection from excitotoxicity. Cannabimimetic drugs did not protect cells from the direct application of glutamate (30 microM). These data suggest that cannabimimetic drugs may slow the progression of neurodegenerative diseases.

Analgesics↗

Neurotoxicity mediated by aberrant patterns of synaptic activity between rat hippocampal neurons in culture.

Reducing the extracellular Mg2+ concentration ([Mg2+]o) to 0.1 mM evoked an aberrant pattern of glutamatergic activity in the synaptic network formed by rat hippocampal neurons grown in primary culture. This treatment resulted in a significant increase in neuronal death when maintained for 20-24 h; 0.1 mM [Mg2+]o elicited a stable and repetitive series of intracellular Ca2+ concentration ([Ca2+]i) spikes as indicated by indo-1-based microfluorimetry. Fura-2-based digital imaging experiments found that the [Ca2+]i spikes were synchronized for all the neurons in a given field. Thus electrophysiological recordings from individual cells were reasonable representations of the field as a whole, enabling correlation of electrical activity to viability. Underlying each [Ca2+]i spike was an intense burst of action potentials. Whole cell voltage-clamp experiments showed that a burst was composed of fast action currents superimposed on a slow inward current. The N-methyl--aspartate (NMDA) receptor antagonist CGS19755 (10 microM) blocked [Ca2+]i spiking, the slow inward current, and the cell death induced by low [Mg2+]o. The L-type Ca2+ channel antagonist nimodipine (10 microM) blocked [Ca2+]i spiking, all synaptic activity, and the cell death induced by low [Mg2+]o. The non-NMDA receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX, 10 microM) exerted variable effects on [Ca2+]i spiking and blocked the slow inward current only when the cells were held at a relatively negative holding potential. CNQX did not afford any protection from 0.1 mM [Mg2+]o-induced neurotoxicity. [Ca2+]i imaging experiments showed that CNQX inhibited [Ca2+]i spiking in a subset of neurons within an active network. Thus, the neurons that were insensitive to CNQX appear to be those that were destined to die. We characterized an in vitro model that allowed us to correlate specific electrophysiological components of glutamatergic synaptic activity to the subsequent viability of the network. A slow NMDA receptor-mediated inward current was required to elicit [Ca2+]i spiking and neurotoxicity. Non-NMDA receptors did not contribute to synaptically mediated cell death in this model. An L-type Ca2+ channel antagonist was neuroprotective when used at concentrations that blocked synaptic activity, suggesting that dendritic L-type Ca2+ channels present a useful target for neuroprotective drugs.

6-Cyano-7-nitroquinoxaline-2,3-dione↗

Mononeuropathy multiplex in hemophilia: an electrophysiologic assessment.

Single nerve lesion or mononeuropathy caused by extrinsic compression of hematoma or hemarthrosis is consistently reported for hemophilic patients. To study the manifestation of peripheral nerve lesions, an extensive nerve conduction study of peripheral nerves and specific nerves in correlation with clinical involvement was carried out on 126 hemophilic patients. We found 24 patients with 34 peripheral nerve lesions. The frequency of nerve lesion positively correlated with the severity of hemophilia (p < 0.01) and increasing age (p < 0.05). Nerve lesions occurred more commonly in lower extremities than in upper extremities. The femoral nerve was most commonly involved. Seven patients were identified to have mononeuropathy multiplex with 2 or more nerve lesions involving more than one extremity. All of these patients had chronic condition. Our findings suggest that mononeuropathy multiplex may occur in hemophilic patients.

Adolescent↗

[The relationship between C-erbB-2 expression with cell proliferative activity and prognosis of nasopharyngeal carcinomas].

UNLABELLED: C-erbB-2 and proliferating cell nuclear antigen (PCNA) were detected by immunohistochemical and in situ hybridization methods in nasopharyngeal carcinomas(NPC) and pericarcinomatous tissues(PCT). Some NPC cases were followed up for more than 5 years. RESULTS: The positive rates of C-erbB-2 protein and C-erbB-2 mRNA expression were 87.8%, and 84.0%, respectively in NPC and 74.6% and 76.5%, respectively in PCT. There was a coexpression of C-erbB-2 protein and mRNA. The significant difference for PCNA staining intensity index(S II) existed in the vesico-nuclear and poorly differentiated types of NPC and in the C-erbB-2 staining cases of NPC. No correlation was found between the expression of C-erbB-2 protein and the clinical stage and metastasis as well as survival rate. CONCLUSION: The C-erbB-2 gene overexpression and cell abnormal proliferation are associated with the carcinogenesis and development of NPC. It is helpful to examine both C-erbB-2 gene expression and PCNA in NPC to evaluate the malignant degree and the effect of radiotherapy.

Carcinoma, Squamous Cell↗

[The factors affecting apoptosis by in situ terminal deoxynucleotide transferase method].

Apoptosis in biopsies of patients with hepatocirrhosis or hepatocellular carcinoma and in small intestinal mucosa of mice were detected by the method of in situ terminal deoxynucleotide transferase (ISTdT). It was found that the best positive result might be obtained by digesting tissue sections with 25mg x L-1 proteinase K for 15 minutes, then treating sections with moderate degree of microwave for 5 minutes after reaction with terminal deoxynucleotide transferase. The best choice of above conditions were discussed.

Animals↗

[The role of cytokeratin 7 in the differential diagnosis of primary ovarian carcinoma and metastatic ovarian carcinoma originated from the gastrointestinal tract].

OBJECTIVE: To assess the role of cytokeratin 7 monoclonal antibody in the differential diagnosis of primary ovarian carcinoma and metastatic ovarian carcinoma originated from the gastrointestinal tract. METHODS: Immunohistochemical study using cytokeratin 7 monoclonal antibody and ABC kit. RESULTS: All the 46 cases of primary ovarian carcinoma were CK 7 positive, while in the metastatic ovarian carcinoma of intestinal origin, all cases remained negative for CK7. Half of the 34 cases of metastatic ovarian carcinoma of gastric origin were CK 7 positive. The positive result of CK7 was significantly higher in the primary ovarian carcinoma than in each group of the metastatic ovarian carcinoma (P < 0.001). CONCLUSION: CK 7 is seemed to be a useful antibody in the differential diagnosis of ovarian carcinoma.

Antibodies, Monoclonal↗

[Epidemiological characteristics of tuberculosis patients complicated with diabetes in Shanghai].

OBJECTIVE: To understand the epidemiological trends and characteristics of tuberculosis patients complicated with diabetes in Shanghai. METHOD: All the registered tuberculosis (TB) cases from 1992 to 1997 in Shanghai were included in the analysis. The new registration rates and percentages of TB cases complicated with diabetes were calculated. RESULT: The new registration rate of the cases increased from 0.70/100,000 in 1992 to 2.10/100,000 in 1997 with an annual increase rate of 24.57%. The cases with the complication accounted for 4.86% in 1997. Diabetes tends to be more prevalent among TB patients in urban area. The greater the age, the more diabetes were found in TB patients. A significant difference was also observed between sex. Bacillary positive rate and percentage of infiltration type were higher, and cavity was more common in cases with the complication. The percentage of cases with relapse was also higher. CONCLUSION: The complication of diabetes significantly affected the epidemiological trend of TB in Shanghai. The diagnosis of diabetes in TB patients should be enforced, and short-course chemotherapy targeting the cases should be investigated.

Age Factors↗

[Analysis of MDMA and its metabolites in urine by GC and GC/MS].

The purpose of this study is to investigate the metabolism of MDMA in the person and establish the methods for the determination of MDMA and its metabolites in urine. MDMA and its metabolite were isolated from urine by liquid-liquid extraction after acidic or enzymatic hydrolysis and were determined by GC/MS(EI, PCI) and GC/FID. The results showed that MDMA was metabolized to MDA and HMMA in man. MDA in the urine after MDMA administration was approximately 0.10-0.14 that of the parent drug. The sensitivity limits of methods were 2-50 ng/ml, and the recoveries were greater than 85%(CV < 10%). It is concluded that the methods were simple, quick and accurate for determining urine concentration of MDMA addicts. MDA/MDMA Ratio can be used to indicate whether the parent drug is taken alone or in combination with its active metabolite.

3,4-Methylenedioxyamphetamine↗

[The application of solid phase extraction for systematic toxicological analysis of abuse drugs].

A new column was developed in this research. It contains a proprietary bonded silica sorbent that exhibits mixed extraction mechanism. A single-column solid-phase extraction procedure was also developed for the screening of acidic, neutral and basic abuse drugs. The recovery of all 7 tested drugs exceeded 60%. The extraction mechanism for different abuse drugs on the new column was explored and was compared with on other columns. It is suggested that this column be effective in systematic toxicological analysis and better than other columns.

Antipsychotic Agents↗

[Abuse of "pheticol" and its interference to the analysis of methamphetamine].

The study was to eliminate interference from ephedrine in the analysis of methamphetamine. The extraction procedure for methamphetamine was modified to include an oxidation step (2 ml urine specimen was treated with 0.5 ml of 1 mol/L phosphate buffer (pH 6.8) and 0.5 ml 0.3 mol/L sodium periodate). Results showed that ephedrine could be oxidized in the presence of periodate ions into smaller fragments while leaving methamphetamine intact. It is recommended that specimens be treated with sodium periodate prior to extraction in order to eliminate any interference caused by ephedrine.

Central Nervous System Stimulants↗

Molecular cloning and characterization of CDEP, a novel human protein containing the ezrin-like domain of the band 4.1 superfamily and the Dbl homology domain of Rho guanine nucleotide exchange factors.

A cDNA for a novel human protein named CDEP was cloned using the subtractive hybridization method between dedifferentiated cartilage cells and overtly differentiated cartilage cells. CDEP cDNA contained an open reading frame encoding 1,045 amino acids in a total length of 3.4 kb. The deduced amino acid sequence revealed that a single polypeptide contained the ezrin-like domain, which is found in cytoskeleton-associated proteins of the band 4.1 superfamily, and the Dbl homology (DH) and pleckstrin homology (PH) domains, which are conserved in the Rho GEF (guanine nucleotide exchange factor) family. Northern blot analysis demonstrated that CDEP mRNA was expressed not only in the differentiated chondrocytes but also in various fetal and adult tissues. Since members of the band 4.1 superfamily and the Rho GEF family are crucial for microfilament organization, the novel protein CDEP may be involved in the adhesion, proliferation, and differentiation of some cell types including chondrocytes via changes in the cytoskeleton.

Amino Acid Sequence↗

Sequence-specific and phosphorylation-dependent proline isomerization: a potential mitotic regulatory mechanism.

Pin1 is an essential and conserved mitotic peptidyl-prolyl isomerase (PPIase) that is distinct from members of two other families of conventional PPIases, cyclophilins and FKBPs (FK-506 binding proteins). In response to their phosphorylation during mitosis, Pin1 binds and regulates members of a highly conserved set of proteins that overlaps with antigens recognized by the mitosis-specific monoclonal antibody MPM-2. Pin1 is here shown to be a phosphorylation-dependent PPIase that specifically recognizes the phosphoserine-proline or phosphothreonine-proline bonds present in mitotic phosphoproteins. Both Pin1 and MPM-2 selected similar phosphorylated serine-proline-containing peptides, providing the basis for the specific interaction between Pin1 and MPM-2 antigens. Pin1 preferentially isomerized proline residues preceded by phosphorylated serine or threonine with up to 1300-fold selectivity compared with unphosphorylated peptides. Pin1 may thus regulate mitotic progression by catalyzing sequence-specific and phosphorylation-dependent proline isomerization.

Amino Acid Isomerases↗