Biomedical subjects
Publications and source records attributed to W He.
Two new compounds were isolated from Isodon pharicus (Prain) Murata. Their structures were determined to be 3 alpha, 11 beta, 13 alpha-trihydroxy-entkaur-16-en-15-one (1), named isodopharicin D, and 11 beta, 13 alpha, 15 alpha-trihydroxy-entkaur-16-en-3 alpha-beta-D-glucoside (2), named isodopharicin F by chemical and spectral evidence.
The use of coupled solid-phase extraction and high-field proton nuclear magnetic resonance spectroscopy (SPE-1HNMR) for the detection and identification of the urinary metabolites of brodimoprim is described. Urine was obtained from Wistar rats following a dose of 200 mg.kg-1 of brodimoprim. Analysis was performed on partially purified extracts obtained by solid-phase extraction onto C-18 bonded silica gel. As each spectrum was often given by a mixture of more than two kinds of metabolites, stepwise identification of the individual metabolites was achieved by sorting out signal sets having invariant proportionality. Five kinds of metabolites of the drug were found by the analysis.
The model of morphine dependent rats was established by gradually increasing the doses of morphine by subcutaneous injection(from 5 mg.kg-1 to 50 mg.kg-1, 5 days). The effects of melatonin(MT) on the mean arterial blood pressure(mABP) and heart rate(HR) were studied in naloxone-precipitated rats. MT was found to inhibit the hypertention and tachycardiac syndromes in morphine withdrawal rats in dose-dependent manner. The concentration of serum norepinephrine(NE) was shown to be decreased by MT. Moreover, regitine was shown to inhibit the increase of mABP in morphine withdrawal rats, but can not inhibit the increase of HR. These results suggest that the inhibitory effect of MT on hypertension and tachycardiac syndromes in morphine withdrawal rats may be mediated by the inhibitory effect of the drug on the sympathetic system.
The effects of pyrazole, which is known to induce hepatic cytochrome P4502A5 (CYP2A5) through posttranscriptional mechanisms, on the level of CYP2A5 in liver and extrahepatic tissues were examined in this study. Intraperitoneal administration of pyrazole at 200 mg/kg for 3 days induced CYP2A4/5 mRNAs and proteins and microsomal coumarin 7-hydroxylation activity in liver and kidney of C57BL/6 mice. A marginal increase (30%) in CYP2A4/5 mRNAs was also observed in the olfactory mucosa but not in the lung, and no increase in CYP2A4/5 proteins or microsomal coumarin 7-hydroxylation activity was observed in either the olfactory mucosa or lung. CYP2A4/5 proteins were not detected on immunoblots in other tissues examined, including breast, bone marrow, testis, prostate, ovary, and uterus from control or pyrazole-treated mice. On the other hand, pyrazole treatment induced CYP2E1 in the olfactory mucosa as well as in liver and kidney, indicating that the olfactory mucosa was exposed to pyrazole. The lack of CYP2A inducibility in the olfactory mucosa was also observed for several other known inducers of hepatic CYP2A5, including cobaltous chloride, stannous chloride, griseofulvin, thioacetamide, and aminotriazole. These results suggest that the mechanisms involved in the induction of hepatic and renal CYP2A5 by pyrazole and other xenobiotic compounds may be tissue-specific.
A high-performance liquid chromatographic (HPLC) method has been developed for the analysis of several benzodiazepines and some of their metabolites in blood, plasma and urine. The method included a liquid-liquid extraction with n-hexane:ethylacetate, a gradient elution on a C8 reversed phase column with a non-electrolyte eluent and a photo diode array detection. This allowed a rapid detection, a purity check, and identification as well as quantitation of the eluting peaks. The detection limit was 10 to 30 ng and the limit of quantitation was 0.05 microgram/mL, using 1 mL of blood, plasma or urine. The procedure is applied routinely in forensic toxicological analyses involving blood, stomach content, urine and organ samples. About 30 positive cases are reported. The avoidance of the use of an electrolyte buffer in the eluent resulted in a robust procedure, free of technical problems and of long rinsing periods, suitable for routine use in forensic toxicology analysis involving blood, urine, stomach content and tissue samples.
We have examined the inducibility of CYP1A1 by beta-naphthoflavone (BNF) in a rat intestinal epithelial cell line, IEC-18, and the associated interaction between cAMP and BNF. CYP1A1 was not constitutively expressed in IEC-18 cells. Upon treatment with BNF, CYP1A1 RNA, protein, and microsomal 7-ethoxyresorufin O-deethylase activity were detected. Treatment with dibutyryl cAMP resulted in a 2-fold increase in the extent of induction at both RNA and protein levels, with corresponding increases in CYP1A1 enzymatic activity. These results support the involvement of protein kinase A in Ah receptor-mediated induction of CYP1A1 and provide an in vitro model for further studies on the mechanisms underlying regional and cellular differences in the regulation of CYP1A1 gene expression in the small intestine.
Endogenous murine angiostatin, identified as an internal fragment of plasminogen, blocks neovascularization and growth of experimental primary and metastatic tumors in vivo. A recombinant protein comprising kringles 1-4 of human plasminogen (amino acids 93-470) expressed in Pichia pastoris had physical properties (molecular size, binding to lysine, reactivity with antibody to kringles 1-3) that mimicked native angiostatin. This recombinant Angiostatin protein inhibited the proliferation of bovine capillary endothelial cells in vitro. Systemic administration of recombinant Angiostatin protein at doses of 1.5 mg/kg suppressed the growth of Lewis lung carcinoma-low metastatic phenotype metastases in C57BL/6 mice by greater than 90%; administration of the recombinant protein at doses of 100 mg/kg also suppressed the growth of primary Lewis lung carcinoma-low metastatic phenotype tumors. These findings demonstrate unambiguously that the antiangiogenic and antitumor activity of endogenous angiostatin resides within kringles 1-4 of plasminogen.
A high performance liquid chromatography (HPLC) assay was developed for the determination of flunitrazepam (FNZ) and its metabolites in urine and plasma. The analytes and the internal standard (triazolam, TRZ) were extracted by Sep-Pak C18 SPE-cartridge and separated utilizing a 5 microm ChromSpher C8 glass column with a gradient mobile phase containing methanol and 0.125% (v/v) of isopropylamine in water. Diode array detection (DAD) was carried out at a monitoring wavelength of 240 nm and a reference wavelength of 550 nm. Standard curves were linear from their quantitation limits until 200 ng ml(-1) urine or 250 ng ml(-1) plasma for 7-amino-desmethyl-flunitrazepam (ADF), 7-amino-flunitrazepam (AF), 7-acetamino-flunitrazepam (ACF) and until 400 ng ml(-1) urine or 500 ng ml(-1) plasma for FNZ, 1-desmethyl-flunitrazepam (DF), and 3-hydroxyl-flunitrazepam (HF). The intraday and interday coefficients of variation ranged from 2.04 to 9.07% and from 2.64 to 14.10%, respectively in urine and from 5.13 to 8.60% and from 7.27 to 10.46%, respectively in plasma. The developed method is used in forensic toxicology and is also applicable to pharmacokinetic studies in man.
Tumour-specific CD4+ T helper (Th) and CD8+ T cytotoxic (Tc) cells may participate in the control and eradication of tumour cells. In the present study, idiotype-specific stimulation of CD4+ and CD8+ blood T cells from patients with monoclonal gammopathy of undetermined significance and patients with untreated multiple myeloma stage I was examined. Activation was measured in the CD4+ and CD8+ subsets enriched by magnetic microbeads as the incorporation of 3H-thymidine and the secretion of interferon (IFN)-gamma, interleukin (IL)-2 and IL-4 by single cells using the enzyme-linked immunospot assay. Idiotype-specific T cells were found in four of seven patients. Stimulation was mainly confined to the CD4+ subset in three of the four responding patients. This type of response was major histocompatibility complex (MHC) class II restricted as it could be inhibited by monoclonal antibodies against MHC class II (HLA-DR), but not against class I (HLA-ABC) molecules. Idiotype-specific CD8+ T cells were also demonstrated in these patients although at a lower frequency. One patient showed a strong and dominating activation of CD8+ T cells which could be blocked by antibodies against HLA-ABC but not against HLA-DR. Idiotype-specific CD4+ or CD8+ T cells were mainly of the type-1 subsets as judged by their secretion of IFN-gamma and IL-2. Thus, this study provides evidence for the presence of idiotype-specific and MHC-restricted CD4+ and CD8+ T cells of the type-1 subsets in patients with monoclonal gammopathies. Such T cells with the potential to control the growth of tumour B cells may be a suitable target for immunotherapeutic interventions in patients.
The objective of the present project was to investigate the efficacy and mechanism of acute (10-minute) adenosine treatment for augmentation of ischemic tolerance in muscle flaps in pigs. Varying doses of adenosine were infused into 28 latissimus dorsi muscle flaps through the axillary artery (0, 0.5, or 2.0 mg per flap) and 22 gracilis muscle flaps through the medial circumflex femoral artery (0, 10, or 20 mg per flap) over 10 minutes. Ten minutes after adenosine infusion, these muscle flaps were subjected to 4 hours of sustained warm global ischemia. In addition, one group of latissimus dorsi muscle flaps (n = 6) received a 10-minute intraarterial adenosine infusion (0.5 mg) at the beginning of reperfusion. Muscle biopsies (n = 4 or 5) for adenosine triphosphate (ATP) analysis were obtained before and after adenosine infusion and at the end of 4 hours of ischemia. The extent of muscle infarction was assessed at 48 hours of reperfusion by the tetrazolium dye staining technique. Muscle blood flow in latissimus dorsi muscle flaps was measured at the end of adenosine infusion (0 or 0.5 mg per flap, n = 8) by the radioactive microsphere (15-microns) technique. It was observed that adenosine, at all doses tested, significantly (p < 0.05) reduced the extent of muscle infarction in latissimus dorsi muscle flaps (control, 40.3 +/- 2.2 percent; 0.5 mg, 20.6 +/- 1.6 percent; 2.0 mg, 18.2 +/- 1 percent) and gracilis muscle flaps (control, 31.0 +/- 1.5 percent; 10 mg, 14.3 +/- 3 percent; 20 mg, 11.6 +/- 1.2 percent). Preischemic adenosine treatment (0.5 mg per flap) was associated with maintenance of a significantly (p < 0.05) higher muscle content of ATP in latissimus dorsi muscle flaps at the end of 4 hours of ischemia compared with saline-treated ischemic controls. Postischemic adenosine treatment did not protect latissimus dorsi muscle flaps against infarction. Furthermore, adenosine treatment did not have any significant effect on mean systemic arterial blood pressure or muscle blood flow in latissimus dorsi muscle flaps. It is concluded that acute (10-minute) preischemic adenosine treatment is effective in augmentation of ischemic tolerance in muscle flaps and that this protective effect of adenosine may be, at least in part, the result of slowing muscle ATP depletion during sustained ischemia. The possible mechanisms of this adenosine-induced energysparing effect are discussed.
We compared the secondary ischemic tolerance of 8 x 12 cm surgically denervated pedicled island skin flaps and skin free flaps raised contralaterally on the buttocks of 50 pigs. The pedicled flaps and free flaps were subjected to 2 hours of primary warm global ischemia followed by 12 hours of reperfusion and 0, 2, 4, 6, or 10 hours of secondary warm global ischemia (n = 10 flaps). Skin necrosis was assessed 7 days after secondary ischemia. Pedicled skin flaps tolerated up to 10 hours of secondary ischemia without skin necrosis. However, incidences of skin necrosis (partial and total) in free flaps subjected to 0, 2, 4, 6, or 10 hours of secondary ischemia were 0, 10, 50, 80, and 100 percent, respectively. In a separate experiment, skin blood flow and hematology were studied in contralateral pedicled flaps and free flaps (n = 20) subjected to 4 hours of secondary ischemia. The skin blood flow measured by 15-micron microspheres at 1.5 hours of reperfusion was significantly higher (p < 0.01, n = 20) in pedicled skin flaps than in skin free flaps (1.91 +/- 0.35 versus 0.67 +/- 0.53 ml/min/100 gm). Under an operating microscope, microthrombi were observed near the arterial and/or venous anastomoses in 8 of 20 skin free flaps but none in the pedicled skin flaps. We obtained venous blood samples by cannulation of the major venae comitantes in 12 of the 20 skin free flaps in which there was no thrombosis in the vascular pedicle for hematologic studies. The venous plasma level of thromboxane B2 was significantly higher (p < 0.05) in the skin free flaps than in their contralateral pedicled skin flaps (195 +/- 49 versus 124 +/- 30 pg/ml). In addition, venous hematocrit, hemoglobin concentration, and white blood cell count also were significantly (p < 0.05) higher in skin free flaps compared with their contralateral pedicled skin flaps. Taken together, these observations were interpreted to indicate that buttock skin free flaps in the pig were less tolerant of secondary ischemia compared with their contralateral pedicled skin flaps subjected to the same ischemic protocol, and this reduced ischemic tolerance in skin free flaps was associated with compromised skin blood flow, hemoconcentration, and thrombosis in the vascular pedicle.
A physical map of Listeria monocytogenes Scott A was generated by the pulsed-field technique of contour-clamped-homogeneous-electric-field (CHEF) electrophoresis. The circular genome of this serotype 4b strain contains 12 AscI fragments (38 to 790 kb), 5 NotI fragments (55 to 1,400 kb), 3 SrfI fragments (110, 1,110, and 2,000 kb), and 2 SfiI fragments (1,320 and 1,920 kb). Summation of individually sized fragments derived by digestion of Scott A genomic DNA with each of these four enzymes provided an average estimated genome length of 3,210 +/- 60 kb. Efforts to assemble the macrorestriction map benefited greatly from the construction and use of pLUCH80 and pLUCH88, temperature-sensitive vectors for delivering transposon Tn917::NotI/SmaI to the chromosome of Scott A. As another component of this study, the positions of four known virulence genes (inlA, mpl, hly, and prf) and three L. monocytogenes-specific sequences (lisM44, lisM51, and lisM52) were localized on the physical map of Scott A by hybridization. Probes prepared from lisM44, lisM51, and the four virulence genes hybridized within a cluster on a 150-kb fragment of the Scott A genome that overlaps part of the NotI-B and AscI-D fragments. The lisM52 probe hybridized with the AscI-F2 (120-kb) fragment of Scott A, which is separated from the NotI-B-AscI-D region by about 300 kb. These results established the first physical and genetic map of a serotype 4b strain of L. monocytogenes and provided further insight on this important food-borne pathogen at the genome level.
We studied the role and mechanism of ATP-sensitive K+ (KATP) channels in ischemic preconditioning (IPC) of skeletal muscle against infarction in vivo. Surgically denervated, noncontractile latissimus dorsi muscle flaps in pentobarbitone-anesthetized pigs were assigned to nine groups: control; IPC (3 cycles of 10-min ischemia/reperfusion); preischemic lemakalim (LMK, 0.18 mg/muscle); postischemic LMK; sodium 5-hydroxydecanoate (5-HD, 27 mg/muscle) before IPC; glibenclamide (Glib 0.3 mg/kg iv) before IPC; 5-HD before preischemic LMK; 5-HD before ischemia; and Glib before ischemia. Except for Glib, all drugs were delivered to each muscle by 10-min local intraarterial infusion to avoid systemic effects. All muscle flaps underwent 4 h of global ischemia. Infarction was assessed at 48 h of reperfusion. In a separate study, muscle biopsies were taken before, during, and after ischemia for assay of high-energy phosphate and lactate contents and myeloperoxidase (MPO) activity. It was observed that muscle infarction in the IPC (24 +/- 2%) and preischemic LMK (21 +/- 2%) groups were smaller (P < 0.05) than that in the control (42 +/- 2%). The anti-infarction effect of IPC and LMK was blocked by 5-HD or Glib. IPC and preischemic LMK caused a higher (P < 0.05) muscle content of ATP and energy charge potential, a lower (P < 0.05) muscle content of lactate during ischemia, and a lower (P < 0.05) muscle MPO activity throughout 16 h of reperfusion compared with the control. These observations indicated for the first time that KATP channels are also involved in the anti-infarction effect of IPC in noncontractile skeletal muscle in vivo. Presently, the cause and importance of energy-sparing and neutrophil-inhibitory effects of IPC and LMK are not known.
We used adenosine A1 receptor agonist N6-1(phenyl-2R-isopropyl)-adenosine (PIA), A1 receptor antagonist 8-cyclopentyl-1,3-dipropylxanthine (DPCPX), and ATP-sensitive K+ (KATP) channel blockers sodium 5-hydroxydecanoate (5-HD) and glibenclamide (Glib), as probes to investigate the role and mechanism of adenosine in ischemic preconditioning (IPC) of noncontractile skeletal muscle against infarction, using the pig latissimus dorsi muscle flap model. Except for Glib, all drugs were delivered to each muscle flap by 10-min local intra-arterial infusion to avoid systemic effects. Muscle flaps that were subjected to 4 h of global ischemia and 48 h of reperfusion sustained 40 +/- 2% infarction. IPC with three cycles of 10 min ischemia and reperfusion, preischemic adenosine, or PIA treatment reduced (P < 0.05) muscle infarction to 24 +/- 2, 18 +/- 2, and 24 +/- 2%, respectively. The anti-infarction effect of IPC and adenosine was blocked by DPCPX, 5-HD, and Glib (P < 0.05). Preischemic adenosine treatment also maintained higher muscle contents of phosphocreatine, ATP, and energy charge potential and lower muscle contents of dephosphorylated metabolites and lactate during ischemia and a lower muscle myeloperoxidase (MPO) activity during reperfusion compared with the control (P < 0.05). Preischemic adenosine treatment did not increase muscle content of adenosine during ischemia or reperfusion. Furthermore, adenosine given at the onset of reperfusion was not effective in attenuating muscle MPO activity or infarction. Taken together, these observations indicate that adenosine, through A1 receptors, initiates the mechanism of IPC with postreceptor involvement of KATP channels in skeletal muscle. However, adenosine is unlikely to play a key role in the effector mechanism. Presently, the cause and role of energy sparing and neutrophil inhibitory effects associated with the anti-infarction effect of preischemic adenosine treatment are unknown.
OBJECTIVE: CD4+ helper T lymphocytes and gamma delta T cells play important role in antimicrobial immunity, however, the mechanism of interactions in question still remains to be elucidated. METHODS: CD4+ T cells and gamma delta T cells separated from peripheral blood lymphocytes of healthy donors by panning technique were stimulated with intracellular pathogen-Mycobacterium tuberculosis and human herpes virus-6 as well as extracellular pathogen-Trichomonas Vaginalis respectively in vitro. RESULTS: The stimuli induced CD4+ T cells significantly to proliferate and producing Th1-type cytokine profile-IL-2 and IFN-gamma. With a semipermeable co-culture system in vitro, antigen-activated CD4+ Th1-like cells were found to promote the proliferation and cytotoxicity of gamma delta T cells via soluble factors. CONCLUSION: Functions of gamma delta T cells might be inhanced by CD4+ cells with Th1-type cytokines in antimicrobial immunity.
OBJECTIVES: To assess the value of B-ultrasonography in the diagnosis and treatment of emergency complications in lung cancer patients. METHODS: Thirty-five patients with lung cancer who suffered from emergency conditions were examined with B-ultrasonography. Of the 35 cases, 5 had superior vena caval syndrome due to compression by a centrally located tumor mass in the right upper lobe; 10 had moderate to large accumulation of fluid in the pericardium with cardiac tamponade; 2 had metastasis to the myocardium and pericardium with acute hemorrhagic pericardial effusion; 20 had massive pleural effusion. The ultrasonographic features of these complications were described. Patients with pericardial or pleural effusion received intervention treatment with Elemen emulsion under B-ultrasound guidance. RESULTS: Combined with systemic chemotherapy, in 4-week period, complications were temporarily ameliorated in 82.8% and became clinically stable in 8.5% of the cases. Treatment was of no effect in 8.5%. CONCLUSION: B-ultrasonography is of value in the diagnosis of emergency complications of lung cancer and is of help to guide intervention treatment.
While producing transgenic mice by ES cell route, it is important to know whether ES cells used have a strong ability to produce chimeras or not. In this report, two methods were used for studying the chimeric ability of two ES cell lines (HDC and MESPU-13). One method was the blastocyst microinjection, that is, chimeras were produced by injecting 15 ES cells into the cavity of C57BL/6J blastocysts. Another was GPI electrophoresis for examinizing the chimerism of ES cells in internal tisssues and organs of chimeras. The results showed that the ability of producing chimeras of MESPU-13 cells was stronger than that of HDC cells. The reason was discussed in detail.
Two compounds were isolated from Isodon pharicus. Their structures were elucidated as 11 beta, 13 alpha, 15 alpha-trihydroxy-entkaur-16-en-3 alpha-(beta-D-glucoside) (named isodopharicin F) and eugenyl-beta-D-glucopyranoside by spectral data and chemical evidence.