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.

Objective: The current study was designed to evaluate the clinical application of radioimmunoimaging(RII) for lymph node metastasis in esophageal carcinoma. Methods:1)131I was used to label McAb G9(specific to cellular membrane antigen of human esophageal carcinoma) and form labeling compound 131I-G9. Administration of 131I-G9 in esophagus submucosally with a specific injector for the purpose of submucosal injection via endoscopies in preoperative patients with squamous cell carcinoma of thoracic esophagus followed by RII. 2)The samples of dissected lymph node were used for detection of radioactivity. Results: 1)The pictures at 48 h showed that small radioactivity concentrated dots appeared dispersedly in mediastinum and upper abdomen around esophagus and cardiac gastric. The lymph nodes were considered metastatic in above regions. 2)The pathological results of the lymph nodes dissected compared to the RII result. The metastatic lymph nodes were found in the regions of dispersedly concentrated radioactivity, while no metastatic lymph nodes could be found in the radioactivity free regions. 3)After counting for radioactivity, lymph node metastases showed higher antibody uptake than the non-metastases lymph nodes. The difference was statistically significant. Conclusion: 131I-G9 may be used to locate metastatic lymph nodes in patients with esophageal carcinoma.

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

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

Objective To investigate the expression of the excision re-pair cross complementing 2( ERCC2) and P53 in the tissue of advanced liver cancer, and the relationship between the expression and the scheme with platinum chemotherapy curative effect and prognosis.Methods A total of 56 cancer patients were given oxaliplatin 135 mg? m-2 and cisp-latin 75 mg? m-2 for treatment of 2-4 recycle.Use the immunohisto-chemistry to detect the expression of the ERCC2 and P53 in the tissue of advanced liver cancer, give a platinum-based drugs chemotherapy, and then evaluate the chemotherapeutic effect, progression -free survival time and overall survival time.Results The expression of ERCC2 and P53 was associated with curative effect of platinum chemotherapy (P<0.05).The overall survival time of patients with positive expression of ERCC2 and P53 was longer than that of patients with the negative expression ( P<0.05 ) , and the progression-free survival time of patients with P53 positives was extended ( P <0.05 ) .Conclusion The expression of the ERCC2 and P53 was associated with the curative effect of platinum chemo-therapy in the gastric carcinoma patients, which can be used as a detection index to evaluate the prognosis of the gastric cancer by chemotherapy.

Objectives To evaluate the expression profile of myoD microRNA-29 (miR-29) family in L6 myoblast differentiated to myotube or L6 myotube treated by glucose and insulin, and to further probe the molecular mechanism of myoD regulating the expression of miR-29 clusters.
Methods The expression of myoD and miR-29 family was detected by using real-time PCR and Western blot analysis. The potential promoter and transcription factors binding sites of miR-29 clusters were predicted by Promoter scan and transcriptional factor search. The promoter sequence of miR-29b1-a and miR-29b2-c cluster was cloned into a luciferase reporter plasmid and the regulatory effect of myoD was analyzed by using dual luciferase reporter assay. Electrophoretic mobility shift assay was further conducted to indicate the binding of myoD on specific sequence. Moreover, overexpression of myoD was achieved by a recombinant adenovirus system (Ad-myoD). L6 cells were infected with Ad-myoD and real-time PCR was conducted to analyze the expression of miR-29b and miR-29c.
Results The expression levels of myoD, miR-29a, miR-29b, and miR-29c were increased in L6 myoblast differentiated to myotube. The expression of myoD, miR-29b, and miR-29c was up-regulated in L6 myotube treated with glucose and insulin, but miR-29a depicted no significant change. Dual luciferase reporter gene assay showed that myoD functioned as a positive regulator of miR-29b2-c expression and myoD could bind to the specific sequence located at the promoter region of miR-29b2-c cluster. Enforced expression of myoD led to a marked increase of miR-29b and miR-29c levels in L6 cells.
Conclusion MyoD might act as a crucial regulator of myogenesis and glucose metabolism in muscle through regulating the expression of miR-29b2-c.

OBJECTIVETo explore the mechanism by which HBV X gene(HBx) inhibits apoptosis of human hepatoma cell line HepG2 in terms of miRNA.

METHODSThree cell lines were prepared: HepG2 cells stably transfected with HBx (HepG2/HBx), HepG2 cells stably transfected with pcDNA3.1 (HepG2/pcDNA3.1) and HepG2 cells. Flow cytometry was adopted to measure the apoptosis of these three cells and Taqman fluorescence quantitative PCR was used to examine miR-192 expression. After HepG2 cells was transfected with miR-192, the apoptosis was analyzed by flow cytometry and the expressions of p53 and PUMA at mRNA and protein levels were evaluated by SYBR Green quantitative PCR and Western blot, respectively.

RESULTSCompared with HepG2/pcDNA3.1 cells (11.46% ± 0.69%) and HepG2 cells (12.5% ± 0.66%), the apoptosis rate of HepG2/HBx cells (2.37% ± 0.35%) was significantly reduced (F = 171.722, P < 0.01). The level of miR-192 was 49.1% ± 5.9% in HepG2 cells, which was dramatically down-regulated (F = 14.319, P = 0.019) as compared to the other two groups (HepG2/pcDNA3.1: 98.0% ± 8.9%; HepG2: 100%). Compared with HepG2 cells transfected with miR-NC (10.74% ± 1.15%), transfection of miR-192 into HepG2 cells led to increased apoptosis (15.74% ± 1.17%) (F = 18.415, P = 0.013) and higher p53 and PUMA expressions at mRNA (p53: 1.68 ± 0.12 vs 0.90 ± 0.09, F = 43.115, P = 0.003, PUMA: 1.66 ± 0.10 vs 0.98 ± 0.06, F = 22.541, P = 0.009) and protein (p53: 3.07 vs 1, PUMA: 2.13 vs 1) levels.

CONCLUSIONHBx could inhibit apoptosis of HepG2 cells through down-regulation of miR-192 which induces apoptosis of HepG2 cells.

Apoptosis ; Carcinoma, Hepatocellular ; genetics ; metabolism ; pathology ; Down-Regulation ; Genes, Viral ; Hep G2 Cells ; Hepatitis B virus ; genetics ; Humans ; Liver Neoplasms ; genetics ; metabolism ; pathology ; MicroRNAs ; metabolism ; Trans-Activators ; genetics ; metabolism