Objective To set up a method for identification of Lonicerae japonicae flos volatile oils using Fourier-transform infrared spectroscopy. Methods The volatile oils of Lonicerae japonicae flos and Lonicerae flos was extracted by steam distillation combined with continuous liquid-liquid extraction with hexane. An oil film was prepared for Fourier-transform infrared spectroscopy scanning by dropping the volatile oils solution on the KBr disc and evaporating the solvent. The obtained infrared spectrum was treated by baseline removing and median filter smoothing. The spectral data within 1800-850 cm-1 was selected as the characteristic spectrum for hierarchical cluster analysis. And the volatile oils of Lonicerae japonicae flos and Lonicerae flos were discriminated by the result of hierarchical cluster analysis. Results Enough volatile oils were extracted for obtaining Fourier-transform infrared spectrum from small amount of Lonicerae japonicae flos. The method developed in the study was able to discriminate Lonicerae japonicae flos volatile oils from Lonicerae flos volatile oils. Conclusion The method can be used for identification of Lonicerae japonicae flos volatile oils.

Objective To explore the role of caveolin-1 in nonalcoholic fatty liver disease (NAFLD) caused by high-fat diet.Methods A total of 12 ten-week-old male C57BL/6 mice were fed with high-fat and high-cholesterol diet for 14 weeks to establish the NAFLD animal model.And six syngeneic mice fed with normal diet at the same time were taken as control.All the mice were sacrificed by the end of 14th week,body weight,liver weight and the changes of serum lipids of the two groups were compared.The changes of caveolin-1 at mRNA and protein levels in the liver of mice with NAFLD were detected by quantitative polymerase chain reaction (qPCR) and Western blot.The liver steatosis of the mice was observed under light microscopy after stained by hematoxylin and eosin.The changes of distribution of caveolin-1 in liver were examined by immunohistochemistry.The differences of caveolin-1 at mRNA and protein level in livers between the two groups were compared by t test.The differences of immunohistochemical scores of caveolin-1 expression in the livers of mice with different degree of fatty liver were analyzed by ordinal variables of two independent samples ranksum test analysis.Results After 14 weeks high-fat and high-cholesterol diet,all the mice of experiment group developed NAFLD.Nine of which were severe and three were moderate.Compared with the control group,serum total cholesterol,high density lipoprotein cholesterol and low density lipoprotein cholesterol of experiment group significantly increased ((1.940 ± 0.300) mmol/L vs (3.771±0.800) mmol/L,(1.120±0.066) mmol/L vs (2.224±0.420) mmol/L,(0.510±0.191) mmol/L vs (1.241±0.660) mmol/L,t=-3.760,-5.474,-3.332,all P＜0.01),however there was no significant difference in triglyceride (P＞0.05).The caveolin-1 of experiment group significantly increased at mRNA (1.536 ±0.226 vs 0.980± 0.272,t=3.371,P＜0.05) and protein levels (0.643±0.240 vs 0.100±0.130,t=4.847,P＜0.01).The immunohistochemical results indicated that the increased caveolin-1 expression mainly distributed in the membrane of hepatocytes,cytoplasm and membrane of lipid droplets.Conclusion The up-regulated caveolin-1 expression in the livers of NAFLD mice induced by high-fat and high-cholesterol may be involved in the mechanism of NAFLD.

Objective To explore the role of glycogen synthase kinase (GSK)-3β/β-catenin signaling pathway on human immunodeficiency virus 1 (HIV-1) negative factor (Nef) protein promoting of human herpesvirus-8 (HHV-8) viral interleukin-6 (vIL-6)-induced angiogenesis.Methods GSK-3β mutant plasmid GSK-3β-S9A,dominant negative (DN) form GSK-3β-DN and the control vector pcDNA3.1+ were transfected into endothelial cells which stably expressed HHV-8 vIL-6 or HIV-1 Nef,or co-expressed vIL-6 and Nef protein.Microtubule formation assay was performed to explore microtubule formation ability.A chick embryo chorioallantoic membrane (CAM) model was used to detect angiogenesis.The expression of GSK-3β/β-catenin signaling pathway-related kinases in transfected cells and CAM tissue were further detected by Western blot.The measurement data were compared by t test.Results The activity of GSK-3β was decreased and the ability of HIV-1 Nef protein was enhanced by transfection with GSK-3β-DN in promoting vIL-6 induced microtubule formation (3.42 vs 2.51,t =3.67,P＜0.01) and angiogenesis (6.25 vs 3.97,t=4.06,P＜0.01).In contrast,the activity of GSK-3β was significantly increased and these functions of HIV-1 Nef protein mentioned above were inhibited by transfection with GSK-3β-S9A (0.62 vs2.51,t=8.48,P＜0.01; 0.39 vs 3.97,t=8.59,P＜0.01).The results of Western blot showed that with the elevated level of,β-catenin (in cells:3.53 vs 2.07,t=6.60,P＜0.05; in tissues:2.76 vs 1.74,t=17.40,P＜0.01) and vascular endothelial growth factor (VEGF,in cells:2.68 vs 1.87,t=4.28,P＜0.01; in tissues:2.20 vs 1.39,t=7.08,P＜0.01) were increased in the GSK-3β-DN transfected cells or tissues,while the opposite results were achieved in the GSK-3β-S9A-transfected cells (GSK-3β phosphorylation:0.50 vs 1.47,t=7.33,P＜0.01; β-catenin:1.05 vs 2.62,t=29.50,P＜0.01; VEGF:0.74 vs 2.16,t=20.95,P＜0.01) or tissues (GSK-3β phosphorylation:0.35 vs 1.97,t=10.72,P＜0.01; β-catenin:0.79 vs 1.77,t=5.72,P＜0.01; VEGF:0.43 vs 1.65,t=11.89,P＜ 0.01).Conclusion GSK-3β/β-catenin signaling pathway is involved in vIL-6-induced angiogenesis promoted by HIV-1 Nef protein,which would be valuable for the therapy of Kaposi's sarcoma,an acquired immunodeficiency syndrome,as a potential molecular target.

OBJECTIVE:To compare the performance of several pattern recognition methods in the identification of volatile oils of traditional Chinese medicine(TCM)by infrared spectroscopy. METHODS:The volatile oils of several Lonicera and Citrus TCM were determined by infrared spectroscopy. All samples of infrared spectrum were classified by hierarchical clustering,K-mean clustering,artificial neural networks,and support vector machine. RESULTS:The results of hierarchical clustering and K-mean clus-tering were ineffective. Methods of artificial neural networks and support vector machine achieved correct classification rate of 100%. CONCLUSIONS:Artificial neural networks and support vector machine can be combined with infrared spectroscopy to cre-ate chemometric fingerprinting for the identification of volatile oils of TCM.

ObjectiveTo reveal status in quo of rehabilitation research according to rehabilitation periodical literature.MethodsMEDLINE and CBM are the retrieval databases during 1995—2002. ICD-10 is the main classification tool of central nervous diseases.ResultsThe percentage of rehabilitation literature is 1.32% on MEDLINE, 1.53% on CBM during 1995—2002. The ratio of rehabilitation literature is 1.67 between MEDLINE and CBM. The common study fields include stroke, paraplegia, hypertension, cerebral palsy, hemiplegia, Parkinson disease, epilepsy, and cerebral infarction, etc.The foreign study takes advantage in multiple sclerosis, and tetraplegia, etc. The fewer fields are neuromyelitis optica, arachnoiditis, and Huntington's disease, etc.ConclusionsThe study of central nervous disease rehabilitation is generally similar on some common CNS diseases, some foreign studies taking advantage.

OBJECTIVETo study the relationship between plasma homocysteine (Hcy) level and deep-vein thrombosis (DVT), and analyze the interaction of Hcy, folate and methylenetetrahydrofolate reductase (MTHFR) gene polymorphism in patients with DVT.

METHODSTotally 69 patients with DVT and 111 healthy controls were included in our case-control study. We determined the MTHFR C677T genotypes by polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP), measured the serum folate and vitamin B12 by radioimmunoassay (RIA), and measured the plasma homocysteine level by fluorescence polarization immunoassay (FPIA).

RESULTSThe frequency of the MTHFR C677T TT genotype had no significant difference between DVT group and control group (P > 0.05). The plasma Hcy level was significantly higher in DVT group than in control group (13.03 +/- 8.74 mumol/L vs 10.14 +/- 4.30 mumol/L, P < 0.05). Both serum folate and VitB12 of patients with DVT were not significantly different from those of controls. The odds radios (OR) of hyperhomocysteinemia for DVT was 2.53 (95% CI 1.08-5.92). The interaction of low folate level and TT genotype increased the risk of DVT (OR = 3.12, 95% CI 1.17-8.38).

CONCLUSIONHyperhomocysteinemia may be an independent risk factor for DVT in Han nationality, while serum folate level and MTHRF C677T genotype are not. An interaction between serum folate level and MTHFR genotype that affect the Hcy level is an important risk factor for DVT.

Adult ; Aged ; Aged, 80 and over ; Female ; Folic Acid ; blood ; Genetic Predisposition to Disease ; Homocysteine ; blood ; genetics ; Humans ; Hyperhomocysteinemia ; blood ; genetics ; Male ; Methylenetetrahydrofolate Reductase (NADPH2) ; genetics ; Middle Aged ; Polymorphism, Restriction Fragment Length ; Venous Thrombosis ; blood ; etiology ; genetics ; Vitamin B 12 ; blood

OBJECTIVETo observe the effect of Huatuo Zaizao extractum (HTZZ) on focal cerebral ischemia/reperfusion (I/R) neurogenesis in rats induced by middle cerebral artery occlusion (MCAO) and its mechanism.

METHODTotally 55 healthy adult male Sprague-Dawley rats were divided into the sham operation group, the MCAO model group and HTZZ high, middle and low dose groups (5, 2.5, 1.25 g x kg(-1)), with 11 rats in each group, and orally administered with drugs. The focal cerebral ischemia model was established by performing a middle cerebral artery occlusion (MCAO, 90 min) followed by a seven-day reperfusion (once a day). The neurogenesis and expressions of extracellular signal-regulated kinase (ERK) and cAMP response element binding protein (CREB) were detected by the immunofluorescent staining. The enzyme linked immunosorbent assay (ELISA) was adopted to determine the vascular endothelial growth factor (VEGF) and brain-derived neurotrophic factor (BDNF).

RESULTMCAO (90 min) followed by a seven-day reperfusion resulted in the significant increase in the number of penumbra cortex newborn neurons (BrdU(+) -NeuN(+)), which was accompanied by the growth of ERK and CREB phosphorylation and VEGF and BDNF levels. HTZZ could promote the generation of newborn neurons (BrdU(+)-NeuN(+)) and the ERK and CREB phosphorylation and increase VEGF and BDNF levels at the ischemic side.

CONCLUSIONHTZZ could promote the neurogenesis, which may be the interventional targets of effective traditional Chinese medicine Huatuo Zaizao extractum in promoting the self-repair function of the cerebral ischemic areas.

Animals ; Brain Ischemia ; drug therapy ; genetics ; metabolism ; physiopathology ; Brain-Derived Neurotrophic Factor ; genetics ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Male ; Neurogenesis ; drug effects ; Neurons ; cytology ; drug effects ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reperfusion ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

OBJECTIVETo investigate the immunoregulation mechanism of Bushen Recipe (BSR) in patients with chronic hepatitis B (CHB) of Gan-Shen yin-deficiency and lingering damp-heat syndrome (GSS).

METHODSThirty-five patients with positive HBV DNA and abnormal alanine transaminase (ALT) level were assigned to the treatment group (22 patients) and the control group (13 patients), they were treated with BSR and alpha-2b interferon for 6 months respectively. Blood levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBIL) and HBV DNA were measured before and after treatment. And the expressions of immune effector molecules of nature killer (NK) cell, including perforin (PF), granzyme B (GrB), granulysin (GNLY), tumor necrosis factor-alpha (TNF-alpha) and gamma-interferon (gamma-IFN), were detected using flow cytometry.

RESULTSLevels of ALT and AST declined significantly in both groups after treatment (P < 0.05 or P < 0.01), showing insignificant difference between them. And the expressions (%) of PF and GNLY in the treatment group reduced significantly after treatment, from 69.62 +/- 27.58 to 34.86 +/- 31.60 for PF and from 64.54 +/- 25.96 to 25.72 +/- 24.98 for GNLY (both P < 0.05). In the treatment group and the control group, as compared with before treatment, the total scores of Chinese medicine symptoms were significantly declined after treatment (P < 0.01), and the total scores of Chinese medicine symtoms in the treatment group was significantly lower than that of the control group (P < 0.05). As compared with the total effective rate of Chinese medicine syndromes in the control group after treatment, that in the treatment group was significantly increased (P < 0.05).

CONCLUSIONThe mechanism of BSR in immuneregulating on patients with CHB of GSS is by way of declining the expressions of relative immune effector molecules to promote the recovery of the damaged liver.

Adjuvants, Immunologic ; therapeutic use ; Adolescent ; Adult ; Diagnosis, Differential ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Hepatitis B, Chronic ; drug therapy ; immunology ; Humans ; Killer Cells, Natural ; immunology ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Phytotherapy ; Young Adult

Objective To evaluate the values of gastrin‐releasing peptide (pro‐GRP) and neuronspecific enolase (NSE) in differ‐ential diagnosing of small cell lung cancer (SCLC) .Methods Serum samples from 120 SCLC patients ,130 non‐small cell lung canc‐er (NSCLC) ,80 Patients with benign lung disease and 90 healthy donors were collected to detect the level of pro‐GRP and NSE .All data were analyzed by SPSS13 .0 and then we analyzed the serum level and positive rates of the two tumor markers .ROC was gener‐ated by GraphPad Prism 5 .Results The expression level of pro‐GRP and NSE in SCLC group were significant higher than NSCLC group、benign lung disease group and healthy donors group (P<0 .05) .The positive rates of pro‐GRP in SCLC were higher than the other three groups (P<0 .05) .However ,there had no significant difference between SCLC group and NSCLC group in the posi‐tive rates of NSE(P>0 .05) .ROC area under curve of pro‐GRP ,NSE and both were 0 .890 ,0 .810 and 0 .915 ,separately .Conclusion The tumor biomarker of NSE could only identify of benign and malignant lung diseases ,but can not identify the type of lung canc‐er including SCLC and NSCLC ;Nonetheless the tumor biomarker of pro‐GRP could not only identify benign and malignant lung dis‐eases ,but also identify the pathological type of SCLC and NSCLC ;Combined determination of pro‐GRP and NSE had significant values for the differential diagnosis of SCLC .

OBJECTIVETo investigate the effects of endothelial microvesicles (EMVs) induced by calcium ionophore A23187 on H9c2 cardiomyocytes.

METHODSHuman umbilical vein endothelial cells (HUVECs) were treated with 10 micromol/L A23187 for 30 min. EMVs from HUVECs were isolated by ultracentrifugation from the conditioned culture medium. EMVs were characterized using 1 and 2 microm latex beads and anti-PE-CD144 antibody by flow cytometry. For functional research, EMVs at different concentrations were cocultured with H9c2 cardiomyocytes for 6 h. Cell viability of H9c2 cells and the activity of LDH leaked from H9c2 cells were tested by colorimetry. Moreover, apoptosis of H9c2 cells was observed through Hoechst 33258 staining and tested by FITC-Annexin V/PI double staining.

RESULTSEMVs were induced by A23187 on HUVECs, and isolated by ultracentrifugation. We identified the membrane vesicles (< 1 microm) induced by A23187 were CD144 positive. In addition, the EMVs could significantly reduce the viability of H9c2 cells, and increase LDH leakage from H9c2 cells in a dose dependent manner (P < 0.05). Condensed nuclei could be observed with the increasing concentrations of EMVs through Hoechst 33258 staining. Furthermore, increased apoptosis rates of H9c2 cells could be assessed through FITC-Annexin V/PI double staining by flow cytometry.

CONCLUSIONMicrovesicles could be released from HUVECs after induced by A23187 through calcium influx, and these EMVs exerted a pro-apoptotic effect on H9c2 cells by induction of apoptosis.

Annexin A5 ; Apoptosis ; Calcimycin ; pharmacology ; Calcium ; metabolism ; Cell Line ; Cell Membrane ; drug effects ; Coculture Techniques ; Flow Cytometry ; Fluorescein-5-isothiocyanate ; analogs & derivatives ; Human Umbilical Vein Endothelial Cells ; Humans ; Myocytes, Cardiac ; drug effects ; Staining and Labeling