Phenylpropanoid glycosides from Sophorae Tonkinensis Radix et Rhizoma / 中国中药杂志

Three phenylpropanoid glycosides were purified by extensive chromatographic techniques including column chromatography over microporous resin, MCI, diol, sephadex LH-20, reverse phase C_(18) from water-extracts of Sophorae Tonkinensis Radix et Rhizoma. Their planar structures were elucidated by combination of various spectroscopic method, such as IR, UV, MS, and NMR. The absolute configuration of aglycone was determined by quantum chemical calculations. Their structures were elucidated as(8R)-3-(4-hydroxyphenyl)-1-propanol-2-O-β-D-glucopyranoside(1), kalopanaxin D(2),(E)-4-hydroxycinnamyl alcohol 4-O-[2'-O-β-D-apiofuranosyl(1″→2')]-β-D-glucopyranoside(3). Compound 1 was undescribed previously. Compounds 2 and 3 were firstly isolated from Sophora genus.

Effect of Nature and Flavor Subdivision of Ephedrae Herba on Rats Model of Harmful Fluid Retention in Upper Jiao / 中国实验方剂学杂志

Objective: To study the effective substance foundation of Ephedrae Herba and explore its mechanism, in order to further enrich the theory of drug resistance of Ephedrae Herba.Method: In this experiment, a compound model was used to establish rat model of Harmful Fluid Retention in upper Jiao. The Rats were randomly divided into model group, captopril group (4.38 mg·kg-1), Ephedrae Herba decoction group(468 mg·kg-1), polysaccharide group (265.36 mg·kg-1), volatile oil group (2.34 mg·kg-1), alkaloid group(40.71 mg·kg-1) and phenolic acid group (210.60 mg·kg-1), and normal group (10 mL·kg-1). The normal group and the model group were given the same volume of normal saline for four weeks. The 24 h urine volume of rats was collected by metabolic cage method. The changes of heart and lung tissue morphology were observed under light microscope. The heart index, lung index, left ventricular ejection fraction(LVEF), left ventricular short axis shortening rate(LVFS) and pulmonary permeability index, number(LPI), lung dry-wet ratio(W/D), creatine kinase isoenzyme(CK-MB), angiotensin Ⅱ(Ang Ⅱ), aldosterone(ALD), cardiac aquaporin 1(AQP1), lung AQP1, aquaporin-3(AQP3) and kidney AQP1, aquaporin-2(AQP2), interleukin-6(IL-6) and tumor necrosis factor-α(TNF-α) change were detected.Result: Compared with the normal group, heart and lungs of the model group were significantly damaged. The amount of 24 h urine, LVEF, LVFS of model rats were significantly reduced(Pα were significantly increased(PPα were significantly increased (PPα were significantly reduced (PPConclusion: Alkaloid components "Wen" and "Xin" are the effective substance basis of its action. The mechanism may be related to the inhibition of renin angiotensin aldosterone system (RAAS) and the anti-inflammatory effect.

Evaluation of estrogenic effects of Semen Descurainiae and its action mechanism / 中国药理学通报

Aim To evaluate the estrogen-like activity of Semen Descurainiae aqueous extracts (SD-ae), to deter-mine its effective chemical separation components and to study the mechanisms. Methods The estrogen-like ac-tivity of SD-ae and its effective chemical separation com-ponents were evaluated by the animal experiment, uterine weight test and cell experiment, namely E-SCREEN ex-periment. Estrogen receptor antagonist ICI182,780 inter-vention blocking experiment was carried out to detect the pathway of estrogen-like action; the HEK293 cells were co-transfected with the report gene carrier and the ERα, ERβ expression vector by cationic liposome, the report gene carrier was constructed via the estrogen-responsive component (ERE) and the report gene luciferase (Luc), then the estrogen-like signaling pathway was evaluated with standardized Luc activity; the expression of estrogen receptor ERα, ERβ and estrogen-induced gene PR mR-NA was detected by real-time fluorescence quantitative PCR. Results Compared with normal control, SD-ae low and high dose could significantly improve the uterine coefficient of immature female mice(P<0.05), and the oligosaccharides composition of Semen Descurainiae aque-ous extracts(SD-ae-Oli) and the polysaccharide composi-tion of Semen Descurainiae aqueous extracts(SD-ae-Pol) also significantly improved the uterine coefficient of im-mature female mice (P<0.01 or P<0.05); SD-ae, SD-ae-Oli and SD-ae-Pol had a significant proliferative effect on MCF-7 cells ( P <0.01 or P <0.05), while ICI182,780 intervened to block its proliferative effect. The reporter gene technology showed that the standardized Luc activities of SD-ae, SD-ae-Oli and SD-ae-Pol were significantly higher than those of the normal control when they were induced by ERβ respectively (P<0.01); and the SD-ae significantly increased the expression of ERβ mRNA in mouse uterus than the normal control, but no effect was found on the expression of ERα and PR mR- NA. Conclusions The estrogenic effect of SD-ae may be found at the first time, and its effective chemical sepa-ration components are SD-ae-Oli and SD-ae-Pol. Their estrogenic effects are mediated by ERβ. The molecular mechanism of the estrogenic effects is probably that SD-ae promotes the expression of ERβ mRNA.

Clinicopathological and prognostic significance of hypoxia-inducible factor-1 alpha in lung cancer: a systematic review with meta-analysis / 华中科技大学学报（医学）（英德文版）

Hypoxia-inducible factor-1 alpha (HIF-1α) plays a vital role in the initiation, evaluation and prognosis in lung cancer. The prognostic value of HIF-1α reported in diverse study remains disputable. Accordingly, a meta-analysis was implemented to further understand the prognostic role of HIF-1α in lung cancer. The relationship between HIF-1α and the clinicopathological characteristics and prognosis of lung cancer were investigated by a meta-analysis. PubMed and Embase were searched from their inception to January 2015 for observational studies. Fixed-effects or random-effects meta-analyses were used to calculate odds ratios and 95% confidence intervals of different comparisons. A total of 20 studies met the criteria. The results showed that HIF-1α expression in lung cancer tissues was significantly higher than that in normal lung tissues. Expression of HIF-1α in patients with squamous cell carcinoma was significantly higher than that of patients with adenocarcinomas. Similarly, non-small cell lung cancer (NSCLC) patients had higher HIF-1α expression than small cell lung cancer (SCLC) patients. Moreover, lymph node metastasized tissues had higher HIF-1α expression than non-lymph node metastasized tissues. A high level HIF-1α expression was well correlated with the expression of vascular endothelial growth factor and epidermal growth factor receptor in the NSCLC. Notably, NSCLC or SCLC patients with positive HIF-1α expression in tumor tissues had lower overall survival rate than patients with negative HIF-1α expression. It was suggested that HIF-1α expression may be a prognostic biomarker and a potential therapeutic target for lung cancer.

Reversing effects of curcumin on multi-drug resistance of Bel7402/5-fu cell line / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To investigate the reversing effects of curcumin on hepatocellular carcinoma drug resistance Bel7402/5-Fu cell line.</p><p><b>METHODS</b>Through the exposure to gradual increased concentrations of 5-fluorouracil (5-Fu), the cell line Bel7402 was induced to establish a multi-drug resistant sub-cell line Bel7402/5-Fu. The sensitivity to 6 chemotherapeutics of Bel7402 and Bel7402/5-Fu were detected using methyl thiazolyl tetrazolium (MTT) assay. The 50% inhibitory concentration (IC50) and resistant index (RI) were calculated. The differences of the inhibition ratio of Bel7402/5-Fu by curcumin, 5-Fu, curcumin combined with 5-Fu were detected using MTT assay. The effects of curcumin, 5-Fu, curcumin combined with 5-Fu on the Bel7402/5-Fu apoptosis were detected using flow cytometry.</p><p><b>RESULTS</b>The Bel7402/5-Fu cell line showed multi-drug resistance (MDR) to various chemotherapeutics, with the highest RI shown of 5-Fu (being 109.55 +/- 14.30 times). The inhibition ratio of 5, 10, and 20 microg/mL curcumin combined with 5-Fu (50% IC50) was respectively 21.47% +/- 1.49%, 27.10% +/- 2.32%, and 59.37% +/- 2.45%. The Bel7402/5-Fu apoptosis ratio of 5, 10, and 20 microg/mL curcumin combined with 5-Fu (50% IC50) was 30.92% +/- 2.10%, 44.87% +/- 2.24%, and 50.36% +/- 2.58%, respectively, which was obviously higher than that of the curcumin group and the 5-Fu group. Besides, the apoptosis rate increased along with increased curcumin concentration in the range of 0 -20 microg/mL.</p><p><b>CONCLUSION</b>Curcumin could induce the apoptosis of Bel7402/5-Fu. Meanwhile, it showed favorable reversing effects on MDR.</p>

Randomized study on the safety and efficacy of dual-axis rotational versus standard coronary angiography in / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Dual-axis rotational coronary angiography (DARCA) was developed as an innovative adaptation of rotational angiography (RA), but it requires a longer coronary injection compared to standard coronary angiography (SA). As the body of the average Chinese patient is smaller than that of most western patients, with the same contrast injection time, the risk of complications from the contrast agent is increased in this population. The purpose of this study was to assess the clinical safety and efficacy of DARCA in the diagnosis of coronary artery disease (CAD) in the Chinese population by directly comparing it to SA.</p><p><b>METHODS</b>Two hundred Chinese patients were randomized to either the SA group (n = 100) or DARCA group (n = 100). Contrast utilization, radiation exposure and procedure time were recorded for each modalities. Blood pressure (BP), heart rate (HR) pre and post injection symptoms and any arrhythmias were recorded.</p><p><b>RESULTS</b>Compared to the SA group, there was a 42% reduction in contrast utilization, 55% reduction in radiation exposure and a 31% shorter procedure time in the DARCA group. In both groups, there were slight declines in the systolic BP values in the left coronary artery (LCA) post injection (P < 0.01). Moreover, post injection HRs for the LCA were also reduced in the DARCA group (P < 0.01). But all of these changes were small, transient and without clinical importance. Only one patient (1%) in the DARCA group had an attack of ventricular tachycardia immediately post injection and it resolved by itself during LCA angiography. No arrhythmias occurred in the SA group.</p><p><b>CONCLUSION</b>DARCA is a safe, efficient, and clinically comparable alternative to SA in the diagnosis of coronary artery disease in the Chinese population with less contrast utilized, which is less radiation exposure and a shorter procedure time than SA.</p>

Long-term effects of biodegradable versus durable polymer-coated sirolimus-eluting stents on coronary arterial wall morphology assessed by virtual histology intravascular ultrasound / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The durable presence of polymer coating on drug-eluting stent (DES) surface may be one of the principal reasons for stent thrombosis. The long-term coronary arterial response to biodegradable polymer-coated sirolimus-eluting stent (BSES) in vivo remained unclear.</p><p><b>METHODS</b>Forty-one patients were enrolled in this study and virtual histology intravascular ultrasound (VH-IVUS) was performed to assess the native artery vascular responses to BSES compared with durable polymer-coated SES (DSES) during long-term follow-up (median: 8 months). The incidence of necrotic core abutting to the lumen was evaluated at follow-up.</p><p><b>RESULTS</b>With similar in-stent late luminal loss (0.15 mm (0.06-0.30 mm) vs. 0.19 mm (0.03-0.30 mm), P = 0.772), the overall incidence of necrotic core abutting to the lumen was significantly less in BSES group than in DSES group (44% vs. 63%, P < 0.05) (proximal 18%, stented site 14% and distal 12% in BSES group, proximal 19%, stented site 28% and distal 16% in DSES group). The DSES-treated segments had a significant higher incidence of necrotic core abutting to the lumen through the stent struts (73% vs. 36%, P < 0.01). In addition, more multiple necrotic core abutting to the lumen was observed in DSES group (overall: 63% vs. 36%, P < 0.05). Furthermore, when the stented segments with necrotic core abutting to the lumen had been taken into account only, DSES-treated lesions tended to contain more multiple necrotic core abutting to the lumen through the stent struts than BSES-treated lesions (74% vs. 33%), although there was no statistically significant difference between them (P = 0.06).</p><p><b>CONCLUSIONS</b>By VH-IVUS analysis at follow-up, a greater frequency of stable lesion morphometry was shown in lesions treated with BSESs compared with lesions treated with DSESs. The major reason was BSES produced less toxicity to the arterial wall and facilitated neointimal healing as a result of polymer coating on DES surface biodegraded as time went by.</p>

Coronary plaque characterization of nonculprit or nontarget lesions assessed by analysis of in vivo intracoronary ultrasound radio-frequency data / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Unheralded sudden death and acute myocardial infarction are common manifestations of coronary atherosclerosis. Such events are related to thrombotic occlusion at the site of non-flow limiting atherosclerotic plaques in epicardial coronary arteries. This study aimed to assess plaque characterization of nonculprit lesions in patients with acute coronary syndrome (ACS) compared with those with stable angina pectoris (SAP) determined by analysis of intravascular ultrasound (IVUS) radiofrequency (RF) data.</p><p><b>METHODS</b>In 81 patients, nonculprit vessels with < 50% diameter stenosis and nontarget segment of culprit vessels with < 50% diameter stenosis were studied with IVUS. Tissue maps were reconstructed from RF data using IVUS-Virtual Histology software.</p><p><b>RESULTS</b>Mean lipid core percentage was significantly higher in patients with ACS than in those with SAP ((25.78 +/- 6.30)% vs (9.11 +/- 4.90)%, P < 0.001). In addition, patients with SAP showed more fibrotic vessels ((59.66 +/- 16.87)% vs (49.07 +/- 10.20)%, P < 0.001). There was no significant difference in either mean calcium ((4.37 +/- 2.40)% vs (5.12 +/- 3.00)%, P = 0.225) or fibrolipid ((24.94 +/- 9.40)% vs (25.82 +/- 13.60)%, P = 0.731) percentages in nonculprit vessels, but the mean calcium percentage was significantly higher in nontarget lesions of culprit vessels ((5.51 +/- 3.29)% vs (3.57 +/- 2.10)%, P = 0.003). In addition, there was a positive correlation between lipid core and remodeling index (RI) (r = 0.847, P < 0.001) and a negative correlation between fibrous tissue and RI (r = -0.946, P < 0.001).</p><p><b>CONCLUSIONS</b>In this study, in both nonculprit vessels and nontarget lesion of culprit vessels, plaque characterization of nonculprit lesions determined by spectral analysis of IVUS RF data was significantly different in patients with ACS. The percentage of lipid core was significantly higher in patients with ACS than in those with SAP. Conversely, SAP patients showed more fibrotic content. In vivo plaque composition and morphological changes were related to remodeling of the coronary artery tree.</p>

Research of signaling pathway of vascular endothelial growth factor regulating by hepatitis B virus X protein / 中华外科杂志

<p><b>OBJECTIVE</b>To study the function of nuclear factor-kappaB (NF-kappaB) signaling pathway in regulating vascular endothelial growth factor (VEGF) by hepatitis B virus X protein (HBx).</p><p><b>METHODS</b>After the establishment of L02-HBx cell line with stable transfected HBx gene, NF-kappaB signaling pathway blocker PDTC was introduced to cut off its signal transduction. Double immunofluorescent staining and laser scanning confocal microscopy were applied to study the activation and deactivation of NF-kappaB signaling pathway. Real-time PCR and Western blot were used to observe the expression of VEGF gene before and after the HBx transfection, as well as the treatment with PDTC.</p><p><b>RESULTS</b>The NF-kappaB signaling pathway of L02-HBx cells was activated after transfection with HBx gene as compared to the control L02 cells without treatment. The mRNA and protein levels of VEGF in L02-HBx cells increased 4.07 +/- 0.31 and 4.34 +/- 0.64 times respectively. The difference was of statistical significance (P < 0.05) in comparison with the control cells. The mRNA levels of VEGF decreased to 2.33 +/- 0.22 and 1.86 +/- 0.18(P < 0.05) and at the same time the expression of VEGF also reduced to 2.52 +/- 0.29 and 2.17 +/- 0.34 (P < 0.05), after treatment with 25.0 micromol/L and 50.0 micromol/L PDTC for 24 h respectively when the NF-kappaB signaling pathway was blocked. There was no significant difference in VEGF mRNA and protein levels when treated with 12.5 micromol/L PDTC for 24 h.</p><p><b>CONCLUSION</b>NF-kappaB signaling pathway maybe one of the routes through which HBx up-regulate the expression of VEGF to promote angiogenesis in hepatocellular carcinoma.</p>

Inducible expression of hypoxia-inducible factor 1alpha on the proliferation and invasion property of HepG2 cells under normoxia in vitro / 中华外科杂志

<p><b>OBJECTIVE</b>To study the inducible expression of hypoxia-inducible factor 1alpha (HIF-1alpha) on the proliferation and invasion property of HepG2 cells under normoxia in vitro.</p><p><b>METHODS</b>Constructed the HepG2(Tet-on-HIF-1alpha) cell line which could induce the expression of HIF-1alpha by doxycycline; Under normoxia in vitro, MTT assay was used to observe the proliferative and adhesive activity of cells, and the invasive activity was determined by transwell cell culture chamber method.</p><p><b>RESULTS</b>Under normoxia, the HIF-1alpha mRNA and protein of HepG2(Tet-on-HIF-1alpha) cells could be induced up to (5.899 +/- 2.176) and (2.179 +/- 0.742) folds by doxycycline (1 microg/ml); There were no difference of A(490 nm) between the Dox(+)and Dox(-) group in experiment detecting the proliferation activity (P > 0.05); But in adhesive experiment, the A(490 nm) of Dox (+) group was 0.662 +/- 0.058, higher than the Dox(-) group 0.526 +/- 0.808 (P = 0.008); The invasive cell number of Dox(+) group was 37.611 +/- 8.424, but in the Dox(-) group, the number was 25.333 +/- 8.117 (P < 0.01).</p><p><b>CONCLUSIONS</b>Under normoxia in vitro, the Tet-on gene regulate system could increase the HIF-1alpha protein by inducing the HIF-1alpha mRNA; HIF-1alpha has no influence with the proliferation activity, but it could enhance the adhesive and invasive properties of HepG2 cells.</p>

Recent advances in metabonomics / 中国医学科学院学报

Metabonomics (or metabolomics) aims at the comprehensive and quantitative analysis of the wide arrays of metabolites in biological samples. Metabonomics has been labeled as one of the new" -omics" joining genomics, transcriptomics, and proteomics as a science employed toward the understanding of global systems biology. It has been widely applied in many research areas including drug toxicology, biomarker discovery, functional genomics, and molecular pathology etc. The comprehensive analysis of the metabonome is particularly challenging due to the diverse chemical natures of metabolites. Metabonomics investigations require special approaches for sample preparation, data-rich analytical chemical measurements, and information mining. The outputs from a metabonomics study allow sample classification, biomarker discovery, and interpretation of the reasons for classification information. This review focuses on the currently new advances in various technical platforms of metabonomics and its applications in drug discovery and development, disease biomarker identification, plant and microbe related fields.

Central Blood Pressure and the Extent of Coronary Artery Disease / 中华高血压杂志

50%.The patients were categorized into as:one-,two-, and three-vessels coronary artery disease group.Central aortic SBP and DBP was measured by cathetarization dur- ing angiography of coronary artery and brachial blood pressure was measured using cuff method.Results Periph- eral SBP,PP and ascending aortic SBP,PP,fractional systolic pressure(FSP=SBP/MAP)were increased and as cending aortic fractional diastolic pressure(FDP=DBP/MAP)was reduced when the diseased coronary vessels were increased(P

Endocytosis of Ca Alginate Nanocapsules by Dendritic Cells and Function Induction / 中国生物工程杂志

The endocytosis/phagocytosis of calcium alginate nanocapsules by peripheral blood derived dendritic cells was confirmed with the use of Quantum dots labeling.Results further demonstrated these nanoparticles could cause the upregulation of HLA-DR expression to induce the maturation of dendritic cells in vitro.Dendritic cells stimulated by nanocapsules covalently loaded with BSA were shown to stimulate the proliferation of self T cells with blank capsules and free BSA as controls,suggesting their potential applications in cancer cell therapy as a new antigen delivery vehicle with a strong adjuvant effect.

Preparation of Calcium Alginate Nanocapsules with Adjuvant Effect / 中国生物工程杂志

A novel reverse emulsification-phase transition process was designed to prepare alginate-based nanocapsules in the present study. Ca alginate nanocapsules with an average hydrodynamic diameter below 300 nm were obtained by adjusting surfactant and alginate concentration. Instrumental analysis observations revealed a morphologically spherical shape with a relatively uniform size distribution without aggregation.?-potential measurements indicated an topically electrostatically negative nature of capsules in neutral aqueous solution. These nanocapsules were proven to have an equivalent potency to stimulate the maturation of dendritic cells originated from CD14 positive cells isolated from human peripheral blood. Protein loading via covalent conjugation was demonstrated feasible with EDC/NHS facilitated reaction between carboxyl and amino groups. These nanocapsules are considered to have important applications as a new adjuvant in cancer cell therapy, vaccine design and targeted drug delivery for active components of protein nature.

Large recombinant protein displayed on filamentous phage surface and its interaction with small molecule / 生物工程学报

Recombinant proteins were expressed as fusions with the phagemid system of pHEN-KM13 and the characteristics and activities of the fusion proteins displayed on the surface of filamentous phagintain the were studiedon abilty. The altered titer of rescued phages from the phagemid system after trypsin treatment indicated the relative quantity of the phages displaying fusion proteins. The rescue phages displayed foreign proteins could keep the bacterial infection ability, while the bald phage without foreign protein displayed on its surface was sensitive to trypsin treatment and lost the bacterial infection ability. To determine the upper limit for filamentous phage display, four recombinant proteins, glutathione-S-transferase and glutathione-S-transferase fused with three various size peptide linkers, were fused to N terminus of capsid protein gp3 and rescued by helper phage KM13. The rescued phages which displayed fused protein with the size of 40kD or less maintain the infection ability. To assay the activity of the phage displayed protein, the known small molecule probe was used in the interaction study with protein incorporated on phage surface. Results showed that the glutathione-S-transferase on phage surface still bound to glutathione specifically. It indicated that the glutathione-S-transferase displayed on phage surface was correctly folded and functionally active. The results demonstrated that it was feasible to use small molecule probes to interact with the protein displayed on phage surface. In turn, the method described here also demonstrated that phage display system could be utilized to investigate the interactions between protein and small molecules.

Preparation of recombinant HIV-TATm-survivin (T34A) protein and its pro-apoptosis activity to cancer cells in vitro / 生物工程学报

As a novel member of the IAP (Inhibitor of apoptosis protein) family, survivin was observed to be expressed in most human cancerous cells. Fusion protein TATm-survivin (T34A) has drawn considerable attention because it is a potential anti-tumor protein that can be transduced into cancer cell with the help of HIV-TAT domain. In this study, the cDNA encoding survivin was cloned by RT-PCR from human breast cancer cell lines B-Cap-37. An expression vector of pRSET-B-HIV-tatm-survivin (T34A) was constructed by PCR after survivin (T34A) was mutated by site-directed mutagenesis. Subsequently, the resultant plasmid was transformed into E. coli BL21 (DE3). Recombinant HIV-TATm-Survivin (T34A) protein was expressed efficiently with 0.5mM IPTG as inducer, reaching a yield of 650mg/liter (as inclusion body) in fermentation culture. The inclusion bodies were solubilized, refolded and purified to a purity of 96% by ion exchange chromatograghy and size-exclusion chromatography. Remarkable effects of the purified recombinant HIV-TATm-Survivin (T34A) on the morphology of cell line SW1990 and B-Cap-37 were observed after being administrated for 4h. MTT assay showed recombinant HIV-TATm-survivin (T34A) protein could inhibit significantly cell proliferation of SW1990 and B-Cap-37 and SSMC-7721 in vitro. Apoptosis rate and cell circle of SW1990 and B-Cap-37 that had been treated with target protein (final concentration 30 microg/mL) were detected with flow cytometry. Results revealed that more than 65% cancer cells were arrested at G1 phase. The study suggested that TATm-survivin (T34A) protein was a hopeful protein drug in the treatment of cancers by facilitating apoptosis of cancer cells. Key words recombinant HIV-TATm-Survivin (T34A), expression and purification, pro-apoptosis bioactivity, SW1990 and B-Cap-37 cancer cell lines

Purification of a ginsenoside-Rb1 hydrolase from Helix snailase / 生物工程学报

Through a combination of twice DEAE chromatography by NaCl stepwise and gradient elution with gel filtration chromatography, a kind of ginsenoside-Rb1 hydrolase from crude Helix snailase was separated. The hydrolase was purified to apparent homogeneity on SDS-PAGE. It was estimated that the purified hydrolase was consisted of four identical subunits with a molecular mass of 110-115 kD by SDS-PAGE and gel filtration chromatography. The Km and Vmax values for ginsenoside-Rb1 were calculated to be 0.790 mmol/L and 10.192 micromol/(min x mg) of protein respectively. The ginsenoside-Rb1 hydrolase could only hydrolyze the glycosidic bond at the C20 position of ginsenoside-Rb1 into ginsenoside-Rd.

Apolipoprotein E gene polymorphisms and risk for coronary artery disease in Chinese Xinjiang Uygur and Han population / 中国医学科学杂志(英文版)

<p><b>OBJECTIVE</b>To examine the relationship between apolipoprotein E (Apo E) gene polymorphism and risk of coronary artery disease (CAD), analyzing association of polymorphism with classical risk factors.</p><p><b>METHODS</b>A total of 124 patients (including 84 Han population and 40 Uygur population) with angiographically verified CAD or myocardial infarction were prospectively evaluated. Data referring to hypertension, diabetes, and tobacco consumption were recorded. The levels of total cholesterol (TC), high density lipoprotein (HDL) cholesterol, Apo A1 and B, and triglycerides (TG) were determined. DNA was obtained from 124 patients and 70 controls. In order to determine Apo E genotypes, DNA was PCR amplified and digested with HhaI. The genetic polymorphism of Apo E is due to three common alleles, epsilon (epsilon) 2, epsilon3, epsilon4, at a single autosomal gene locus. These alleles determine the six phenotypes E2/2, E3/3, E4/4, E4/2, E4/3, and E3/2.</p><p><b>RESULTS</b>In Uygur population, the frequency of the epsilon2, epsilon3, and epsilon4 was 0.155, 0.648, and 0.197 respectively. In Han population, the frequency of the epsilon2, epsilon3, and epsilon4 was 0.081, 0.772, and 0.146 respectively. In the patient group, the frequency of the epsilon2, epsilon3, and epsilon4 was 0.060, 0.758, and 0.182 respectively. In the control group, the frequency of the epsilon2, epsilon3, and epsilon4 was 0.193, 0.671, and 0.136 respectively. epsilon2 frequency of Uygur' patients and controls was 0.050 and 0.290 respectively. Serum low density lipoprotein (LDL) cholesterol, TC, and TG values tended to decrease from the Apo E-4 phenotypes to Apo E-2 phenotypes. When deletion polymorphism of epsilon2 was compared with the common risk factors for CAD, its risk ratio (RR) is 4.38.</p><p><b>CONCLUSIONS</b>These studies confirm and find that Apo E phenotype distribution in Uygur population differs significantly from that in Han population in Xinjiang. CAD patients have significantly lower epsilon2 allele and slightly higher epsilon3 or epsilon4 allele frequency than controls, especially in Uygur population. It shows protective effects of epsilon2 on CAD.</p>

HRCA and application in detection of genetically modified plant / 生物工程学报

In this article primary studies of the application of hyperbranched rolling cycle amplification (HRCA) in exogenous genes detection of transgenic plants were done. Four padlock probes were designed according to the sequences of four genes/DNA fragments that are used widely in transgenic plants; part of the sequence of pKK233 was chosen as the linking part of padlock probes and a pair of HRCA primers was designed according to the sequence of linking part. Study of the specificity of ligation in HRCA with isotope labeled padlock probes indicated padlock probes could be ringed effectively only when corresponding target DNA exited in the same reaction system and could not be ringed when there was no corresponding target DNA exited. Ligation time is very different according to the characteristic of target DNA being used. 5 min to 10 min is enough if the target DNA is plasmid; 30 min to 60 min is needed if the target is genome DNA of plant because it's sequence is more complex than that of plasmid's. HRCA time was analyzed which indicated longer reaction time can obviously increase the amount of products. Quantity of enzyme in HRCA was also analyzed. Different amount of enzyme (from 0.5 unit to 4 units) can give similar result when other conditions are not changed. On the basis of the research, transgenic tobacco was detected with these four padlock probes and the results were just as prospective. In order to increase the efficiency of detection, multiplex HRCA (MHRCA)was used. In MHRCA more than one padlock probes are used at the same time in the same reaction system to detect more than one targets. Because the amplification products of MHRCA will be complex and it is almost impossible to analyze with electrophoresis, so reverse-blot is used. Detection results of transgenic tobacco with this method are the same with anticipation. Compare to MPCR method we established before MHRCA is more convenient to operate and more effective in detecting exogenous genes in transgenic plants.

Study of the expression membrane protein LASS2 / 生物工程学报

A human membrane protein LASS2 (Homo sapiens longevity assurance homologue 2 of yeast LAG1), which has important physiologic functions, was expressed in three different expression systems. Only the LASS2 protein carboxyl terminal hydrophilic fragment could be expressed in the prokaryote expression system and its polyclonal antibody was produced. The full length of LASS2 protein could be expressed successfully in both eukaryotic in vitro translation system and Bacuvirus expression system: Bac-to-Bac system. SDS-PAGE analysis revealed that the molecular weight of expressed product of LASS2 was about 28 kD. The product was also proved by Western blot. This recombinant LASS2 protein was purified by metal affinity resin and the purity is above 90%.