OBJECTIVE:To optimize the ultrasonic extraction process of total alkaloids from mulberry leaves. METHODS:Based on the single factor experiment,response surface method was adopted to investigate the effects of ultrasonic extraction time, ultrasonic power,ethanol volume fraction,solid-liquid ratio and pH of solvent on the extraction rate of total alkaloids from mulber-ry leaves,then test data were analyzed by Design Expert 8.0.5 software,and the optimized process was confirmed and verified. RE-SULTS:The multiple correlation coefficient of established binomial equation fitting model was 0.969 9;the optimized condition for extracting alkaloid from mulberry leaf was ultrasonic extraction time of 48 min,ultrasonic extraction power of 800 W,ethanol vol-ume fraction of 70%,material-liquid ratio of 1∶25,pH 5. Under these conditions,the extraction rate of total alkaloids was 0.422%, with the bias ratio was less than 2% compared with the model predictions. CONCLUSIONS:The established model has good fit-ting performance,the extraction process is stable and reliable,and can be used for the extraction of alkaloids from mulberry leaves.

OBJECTIVETo explore the relationship between blood stasis (BS) syndrome and coronary lesion in patients with coronary heart disease (CHD).

METHODSSyndrome types of 500 patients collected from multiple centers whose diagnosis of CHD confirmed by coronary angiography were differentiated. And the relationship between BS syndrome, its subtypes, and coronary lesion (affected branches, degree of constriction) were analyzed.

RESULTSThe affected branches of coronary artery in patients of BS syndrome was 2.28 +/- 0.28, while that in the non-BS syndrome patients was 2.07 +/- 0.86, showing significant difference between them (P < 0.05); as compared to patients of non-BS syndrome, the coronary lesions in patients of BS syndrome were mostly multi-vascular, and of more severe degree (P < 0.05). In patients of various BS syndrome subtypes, the average number of affected coronary branches in patients of yang-deficiency subtype was 2.58 +/- 0.65, which was significantly more than the number in patients of other BS syndrome subtypes. The constriction degree of coronary lesions in patients of yang-deficiency BS syndrome subtype were mostly severe or moderate, and single branch lesion was rarely seen, as compared with those in patients of phlegm-stasis obstruction subtype, the difference was significant (P < 0.05). The corresponding correlative analysis showed that close correlation was found between yang-deficiency subtype of BS syndrome and severe coronary constriction with the correlation distance of 0.1899.

CONCLUSIONRelationship between BS syndrome and coronary lesion (its number of branches and degree of constriction) truly exists to a certain extent.

Adult ; Aged ; Coronary Angiography ; Coronary Disease ; diagnosis ; diagnostic imaging ; Coronary Thrombosis ; diagnosis ; diagnostic imaging ; Female ; Humans ; Male ; Middle Aged

Objective To evaluated the effect of hyperbaric oxygenation therapy(HBO)on the RhoA ex- pression and nerve function after transient focal cerebral ischemia in a rat model of middle cerebral artery occlusion. Methods One hundred and twenty-six healthy Sprague-Dawley rats were used and randomly divided into a sham op- eration group(shame group,n=42),a treatment group(n=42),and a control group(n=42).The animal model of middle cerebral artery occlusion(MCAO)was established by using the Zea-Longa method with the animals in the treatment and the control groups,and sham operation was performed with those in the sham group.HBO was applied to the animals in the treatment group.The RhoA protein expression was observed by using immunohistochemistry technique,and the neurological function was evaluated by Bederson's scale at different time points after MCAO.Re- sults(1)Weakly positive expression of RhoA could be located in bilateral cortex and the basal ganglia in the sham group.The expression of RhoA in the treatment group and control group was increased as early as 6 hours after MCAO when compared with that of the sham group,and peaked at 48 h after MCAO and decreased after then,but was still higher than that of the shame group at 7th day to 14th day after MCAO.It was also found that the expression of RhoA of the treatment group was significantly lower than that of the control group(P

Objective：To study the toxicity mechanism(s) of ifosfamide(Ifo) in suspending cultured rat hepatocytes.Methods：Hepatocytes of adult rat were isolated using two-step perfusion method and cultured suspendingly. Cell viability,intracellular enzyme leakage, contents of sulfhydryl groups and MDA contents of hepatocytes were examined 3 hours after ifosfamide was administered at 5,10,20 mmol/L. Surface and ultrastructure of hepatocytes were also observed. Results：Cell viability and TSH,NPSH,PSH contents of hepatocytes significantly declined, and LDH,AST activities in media increased due to the leakage of intracellular enzymes. The decrease in PSH content was ascribed to depletion of TSH. The higher the dose was, the more serious these changes became. However, MDA contents of the hepatocytes were not found increased at any ifo dose groups. In pathological examination, “bulla" formation was found on the surface of the hepatocytes, deformation，swelling even vacuolation of mitochondria and dilation of rough，smooth endoplasmic reticulum were also observed. Conclusions：Ifo has toxic effect on suspending cultured rat hepatocytes. The decrease in sulfhydryl groups contributes to the hepatotoxicity induced by Ifo.

Antineoplastic Agents ; therapeutic use ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; genetics ; Drug Delivery Systems ; Erlotinib Hydrochloride ; Female ; Genes, erbB-1 ; Humans ; Lung Neoplasms ; drug therapy ; genetics ; Male ; Mutation ; Protein Kinase Inhibitors ; therapeutic use ; Quinazolines ; therapeutic use ; Receptor, Epidermal Growth Factor ; antagonists & inhibitors ; genetics

OBJECTIVETo see the change of capillary of heart in Athlete's Heart, so that to discover the mechanism of pathologic change.

METHOD18 male SD rats were separated randomly into control group (without any exercise), aerobic exercise group (swimming for 75 min every day), and overload group (swimming for 180 min with 5% weight of its body every day). After 5 days per week, 12 weeks, exercise training stopped and heart of rats were observed under Transmission Electron Microscope.

RESULTSIn aerobic exercise group, the capillary cavities in heart expand, the walls of capillary become thick; the number of mitochondrion increases; endothelium cells become active in growth. However, after overload exercise, the walls of capillary cockle and protuberances appear. The mitochondrion swell and the cristae become disorder. Most of endosomes expand and their number increases. The karyons become abnormity in shape and uniformity in electronic density, besides the nuclear envelope cockle. The basilar membranes become thick and unclear.

CONCLUSIONAfter exercise training, both physical and pathologic changes in heart capillary are found. In suitable exercises group, the capillaries change physically; the pathologic changes are becoming visible after overload exercise however.

Animals ; Capillaries ; ultrastructure ; Cardiomegaly ; etiology ; pathology ; Male ; Physical Conditioning, Animal ; adverse effects ; Physical Endurance ; Rats ; Rats, Sprague-Dawley ; Sports

OBJECTIVETo investigate the effect of heme oxygenase-1 (HO-1) expression on cell growth and apoptosis in imatinib resistant chronic myeloid leukemia (CML) cells (K562/A02-IM), and explore the relationship between HO-1 gene and CML.

METHODSThe expression of HO-1 in 20 drug-resistant CML patients was detected by RT-PCR. Different concentrations of hemin were used to induce HO-1 expression of K562/A02-IM, HO-1 expression at different time was detected by RT-PCR and Western blot analysis. Cell apoptosis was detected by Annexin V/PI staining, and MTT assay was used to detect viability of K562/A02-IM cells after induction or inhibition of HO-1 gene by hemin and zinc protoporphyrin (ZPP).

RESULTSRT-PCR showed that HO-1 was expressed in the bone marrow mononuclear cells (BMMNCs). When treated with hemin at different concentrations (0, 10, 20, 40 µmol/L) for 16 h, the expression of HO-1 in K562/A02-IM was increased in a dose-dependent manner, and peaked at 20 µmol/L of hemin for 16 h. The apoptosis rates were (17.61 ± 0.01)%, (12.13 ± 0.11)%, (7.94 ± 0.03)% and (4.62 ± 0.15)% at 0,10, 20 and 40 µmol/L of hemin respectively for 16 h and were (14.7 ± 0.05)%, (8.1 ± 0.07)% and (16.3 ± 0.13)% at 20 µmol/L of hemin treatment for 8,16, and 24 h respectively. Hemin induced apoptosis of K562/A02-IM cells in a dose-dependent manner. The expression of HO-1 was induced in K562/A02-IM cells in a dose-dependent manner, and the survival of K562/A02-IM cells was significantly increased as compared to that of control group. When HO-1 was inhibited by ZPP, the cells survival was sharply decreased compared to that of the control group (P < 0.05).

CONCLUSIONHO-1 was expressed in the BMMNCs. It is a kind of molecules whose expression can be induced and can promote the growth of drug-resistant cells. Inhibition of HO-1 expression probably be used for the treatment of drug-resistant CML.

Antineoplastic Agents ; pharmacology ; Benzamides ; Cell Cycle ; Cell Proliferation ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; genetics ; Heme Oxygenase-1 ; genetics ; Humans ; Imatinib Mesylate ; K562 Cells ; Piperazines ; pharmacology ; Pyrimidines ; pharmacology

OBJECTIVEThis study is to see the pathologic change of cardiac myocyte in Athlete's Heart, and explore the mechanism of the pathologic change.

METHODSFifteen male SD rats were separated randomly into control group (without any exercise), aerobic exercise group (Ae group, swimming for 75 min every day), and overloading exercise group (Oe group, swimming for 180 min with a loading of 5 percent of body weight every day). After 5 days per week for 12 weeks, swimming stopped, the rat hearts were prepared to specimens and examined under Transmission Electron Microscope.

RESULTSThe Ae group, the number and volume of mitochondria increased, and the membrane of mitochondria remained entire. Few of dense bodies were found in cytoplasm. The nucleus envelopes of expansion nucleus appear as dentition. These changes were considered as the adaptation to exercises. At the same time, some pathologic changes of the cardiac myocytes similar to senescence also appeared, such as mitochondria expanse, the crista disorder or disappearance, unclear mitochondria membrane, many dense bodies in cytoplasm, nucleus disfiguration and chromatin collection at edge.

CONCLUSIONAfter exercise training, some pathologic changes of cardiac myocyte also occur with physiological changes. With the raise of exercise intension, the pathologic changes become more obvious, even appearance of cardiac myocyte death.

Animals ; Cardiomegaly ; etiology ; pathology ; Exercise Tolerance ; Male ; Microscopy, Electron ; Mitochondria, Heart ; ultrastructure ; Myocardial Contraction ; Myocytes, Cardiac ; ultrastructure ; Rats ; Rats, Sprague-Dawley ; Swimming

OBJECTIVETo define the differences in gene expression patterns between glycidyl methacrylate (GMA)-transformed human lung fibroblast cells (2BS cells) and controls.

METHODSThe mRNA differential display polymerase chain reaction (DD-PCR) technique was used. cDNAs were synthesized by reverse transcription and amplified by PCR using 30 primer combinations. After being screened by dot blot analysis, differentially expressed cDNAs were cloned, sequenced and confirmed by Northern blot analysis.

RESULTSEighteen differentially expressed cDNAs were cloned and sequenced, of which 17 were highly homologous to known genes (homology = 89%-100%) and one was an unknown gene. Northern blot analysis confirmed that eight genes encoding human zinc finger protein 217 (ZNF217), mixed-lineage kinase 3 (MLK-3), ribosomal protein (RP) L15, RPL41, RPS 16, TBX3, stanniocalcin 2 (STC2) and mouse ubiquitin conjugating enzyme (UBC), respectively, were up-regulated, and three genes including human transforming growth factor beta inducible gene (Betaig-h3), alpha-1,2-mannosidase 1A2 (MAN 1A2) gene and an unknown gene were down-regulated in the GMA-transformed cells.

CONCLUSIONAnalysis of the potential function of these genes suggest that they may be possibly linked to a variety of cellular processes such as transcription, signal transduction, protein synthesis and growth, and that their differential expression could contribute to the GMA-induced neoplastic transformation.

Air Pollutants, Occupational ; toxicity ; Carcinoma, Squamous Cell ; genetics ; pathology ; Cell Line, Transformed ; Epoxy Compounds ; toxicity ; Fibroblasts ; cytology ; drug effects ; Gene Expression Profiling ; Glycoproteins ; metabolism ; Humans ; Lung ; cytology ; Male ; Mannosidases ; drug effects ; metabolism ; Methacrylates ; toxicity ; Mitogen-Activated Protein Kinase 3 ; drug effects ; metabolism ; Oligonucleotide Array Sequence Analysis ; Ribosomal Proteins ; metabolism ; Signal Transduction ; genetics ; Transforming Growth Factor beta ; drug effects ; metabolism ; Ubiquitins ; metabolism ; Zinc Fingers ; drug effects ; physiology

OBJECTIVETo evaluate the genotoxic and nongenotoxic effects of short-term exposure to glycidyl mathacrylate (GMA) on human lung fibroblast cells (2BS cells) in vitro.

METHODSDNA strand breakage was determined by single cell gel electrophoresis, and DNA ladder formation assay and flow cytometric analysis were carried out to detect apoptic responses of cells to GMA exposure. The HPRT gene mutation assay was used to evaluate the mutagenicity, and the effect of GMA on gap junctional intercellular communication (GJIC) in the exposed cells was examined with the scrape loading/dye transfer technique. The ability of GMA to transform 2BS cells was also tested by an in vitro cell transformation assay.

RESULTSExposure to GMA resulted in a dose-dependent increase in DNA strand breaks but not apoptic responses. GMA was also shown to significantly induce HPRT gene mutations and morphological transformation in 2BS cells in vitro. In contrast, GMA produced a concentration-dependent inhibition of GJIC.

CONCLUSIONSGMA elicits both genotoxic and nongenotoxic effects on 2BS cells in vitro. The induction of DNA damage and gene mutations and inhibition of GJIC by GMA may casually contribute to GMA-induced cell transformation.

Cell Communication ; Cell Differentiation ; Comet Assay ; DNA Damage ; DNA Mutational Analysis ; Epoxy Compounds ; toxicity ; Fibroblasts ; Gap Junctions ; Humans ; Hypoxanthine Phosphoribosyltransferase ; genetics ; Lung ; cytology ; Methacrylates ; toxicity