以影撷史者:阿素托史·哥瓦力克导演研究

阿素托史·哥瓦力克(Ashutosh Gowariker)于1964年出生于印度孟买,是印度著名导演、演员、编剧和制片人。他以演员的身份开始了职业生涯,一度活跃于印度电视和电影界,经过十余年的历练后转做电影导演。从1993年初执导筒至今,阿素托史以导演兼编剧的身份共推出了9部电影。《印度往事》(Lagaan:Once Upon a Time in India,2001)的成功使他蜚声海外,也拓宽了他的电影创作道路。

关于反对跨境拐卖越南妇女儿童的调研报告

The anticipating aim of the subject is to investigate the illegal cross-border marriage of trafficking Vietnamese women as wives as well as the criminal offence of forcing the Vietnamese women and children to prostitute. By the fieldwork of the Transfer Center for overseas trafficked women and children of Dongxing Public Security Bureau,Guangxi and the so-and-so county in the province,the subject will disclose the criminal chain of forcing the Vietnamese women and children to prostitute from the illegal cross-border marriage of trafficking Vietnamese women as wives.

硫链丝菌素可提高变青链霉菌对阿霉素的耐受性

硫链丝菌素(Thiostrepton,Thio)是一种分离自S.azureus的多肽类抗生素,分子量1650。因为绝大多数放线菌对Thio均非常敏感,所以链霉自遗传工程所构建的重组载体多含有Thio耐受基因(tsr)作为筛选标志。我们在利用p^1J922作为重组载体,变青链霉菌作为宿主克隆阿霉素生物合成基因的研究中首先发现,变青链霉菌和转化有质粒的变青链霉菌孢子对阿霉素的耐受性分别为5ug/ml和10ug/ml。在存在Thio的情况下,上述菌株对阿霉素的耐受性提高一倍。产生这一现象的原因尚不清楚。利用阿霉素产生菌DNA在体外与Thio和阿霉素反应后进行琼脂糖电泳发现:Thio自身对DNA电泳模式无任何影响,但它可明显抑制阿霉素在体外对DNA的嵌合效应。

Doxorubicin Stealth Liposomes Prepared with PEG-Distearoyl Phosphatidylethanolamine and Distribution as well as Antitumor Activity in Mice

目的：研制出能够逃避体内网状内皮细胞的阿霉素隐形脂质体，并考察其在生物体内的分布以及比较阿霉素隐形脂质体与阿霉素普通脂质体的抗肿瘤活性。方法：将聚乙二醇－二硬脂酰磷脂酰乙醇胺（PEG－DSPE）同磷脂酰胆碱和胆固醇材料加在一起，用硫酸铵梯度法制备阿霉素隐形脂质体，同法制备阿霉素普通脂质体（但脂膜中不含PEG－DSPE）；通过尾静脉注射给药，比较隐形脂质体阿霉素、普通脂质体阿霉素和游离型阿霉素（盐酸阿霉素注射液）给药后在小鼠各主要脏器组织和血液中的分布情况；采用动物移植性肿瘤实验法，用H22小鼠肝癌细胞接种于小鼠右侧腋皮下形成实体瘤，考察阿霉素隐形脂质体和普通脂质体给药后对实体瘤的瘤重抑制率。结果：通过硫酸铵梯度法制备出了包封率高达95％的阿霉素隐形脂质体；同游离型阿霉素和普通脂质体阿霉素相比，隐形脂质体阿霉素在血液中浓度显著提高，循环时间显著延长，在心脏中分布的浓度显著降低；按5mg·kg^-1剂量治疗，第二天给药和第七天给药治疗方案，阿霉素隐形脂质体给药组的瘤重抑制率均显著高于普通脂质体给药组的瘤重抑制率；按10mg·kg^-1剂量治疗，隐形脂质体给药组的瘤重抑制率比普通脂质体给药组的瘤重抑制率稍高。结论：用普通脂质体给药相比，隐形脂质体阿霉素给药后延长了其在小鼠血液中的循环时间，说明经过PEG－DSPE修饰后的脂质体有逃避网状内皮细胞吞噬的功能（隐形），并且隐形脂质体阿霉素的抗肿瘤活性显著地提高。

Study on Rational Breeding of High-Yield Avermectin Industrial Strain

[Objective] Strain Biok Av-023 used as the control was employed on screening of high-avermectin yield mutants by rational screening.[Method] With Biok Av-023 as the original strain,the positive mutation strain was firstly screened by routine UV mutagenesis,and then the high-yield avermectin producing strain was selected by the breeding way inferred by L-Ile induction.[Result] UV mutation and L-Ile directional screening had showed that the best L-Ile screening concentration was 0.5%,and the high-yield mutation strain AV60s-32 after re-screening reached the highest titer of 4520 IU/ml,which increased by 23.4% compared with the original strain.[Conclusion] The production of avermectin can be effectively enhanced by the combined way of UV mutation and L-Ile rational breeding.

Analgesic Effects of Tetrodotoxin Combined with Acetylsalicylic Acid on Acetic Acid-induced Abdominal Constriction and Formalin Test in Mice

Aim To study the effects of tetrodotoxin （TTX） combined with acetylsalicylic acid （ASA） on nociceptive stimulus in mice. Methods To assess the antinociceptive effects of TTX, ASA or TTX plus ASA, the acetic acid-induced abdominal constriction test and formalin pain test were used. Results TTX （0.5 - 4.0 μg· kg^-1 ） or ASA （25 - 200 mg· kg^-1 ） im produced a significant inhibition of acetic acid-induced abdominal constriction. The median inhibitory doses （ID508） were 2.1 μg· kg^-1 for TTX（ and 64 mg· kg^-1 for ASA. TTX and ASA also showed a dose-dependent inhibition of the second phase response in the formalin pain model, the ID508, being 2.3μg·kg^-1 and 74.2 mg· kg^-1, respectively. The ihteraction between TTX and ASA was synergistic, as evidenced by the fact that （1） when ASA alone compared with the combination of TTX （0.79 μg · kg^-1 or 0.39μg· kg^-1 ） and ASA, the ID508, of ASA reduced from 64.0 mg· kg^-1 to 5.8 mg· kg^-1 or 12.6 mg· kg^-1, and from 74.2 mg· kg^-1 to 7.4 mg· kg^-1 or 13.0 mg· kg^-1 on tile two models of nociceptive tests, respectively; and that （2） synergism in the analgesic effects was shown by isobiolographic analysis. Conclusion TTX, ASA and the combination of the two drags produce analgesic effects in acetic acid-induced abdominal constriction test and formalin-induced pain test. The interactions between TTX and ASA may be useful in developing novel analgesic agents.

In vitro characterization of two new doxorubicin liposomes modified with argine-glycine-aspartic acid tripeptide or glycine-argine-glycine-aspartic acid-serine pentapeptide

Aim Peptides as ligands have shown the active targeting properties to the receptors like integrins, a family of receptors over-expressed in cancers. The present study was to develop and characterize two peptides modified drug-containing liposomes. Methods Argine-glycine-aspartic acid （RGD） tripeptide and glycine-argine-glycine-aspartic acid-serine （GRGDS） pentapeptide were used for modifications on the doxorubicin-loaded sterically stabilized liposomes （SSL-doxorubicin） for the liposome preparation, RGD-SSL-doxorubicin and GRGDS-SSL-doxorubicin, respectively. Characterizations were performed by measurements of the encapsulation efficiency, particle size and zeta potential, release rates in a simulated in vivo environment, and cytotoxicity to ovarian cancer cells. Cell uptake was investigated by flow cytometry and confocal microscopy methods. Results All encapsulation efficiencies of the liposomes were above 95%, and the modifications using RGD or GRGDS did not affect the final encapsulation efficiency. Average particle sizes of the liposomes Were in the range between 105.7 ± 3.5 nm and 130.5 ± 3.0 nm, and zeta potential values were between -3.3 ± 0.3 and -6.1 ± 0.3 mV. Approximately 2/5 of doxorubicin was released from liposomes before 12 h in the simulated in vivo environment containing fetal bovine serum. Inhibitory rates to cancer cells of the modified liposomes were slightly lower as compared to free doxorubicin. Similar phenomena were observed in the uptake measured by flow cytometry and confocal assay. After uptake applying various formulations on the cancer cells, doxorubicin was mainly distributed in the nuclei of SKOV-3 cells. Conclusion Two new doxorubicin-contained liposomes were successfully prepared and modified with argine-glycine-aspartic acid （RGD） tripeptide and glycine-argine-glycine- aspartic acid-serine （GRGDS） pentapeptide. In vitro characterization indicated that modifications did not alter significantly the properties of the sterically stabilized liposomes.

Studies on Adriamycin Magnetic Gelatin Microspheres

The preparation and properties of adriamycin magnetic gelatin microspheres(Adr- MG-ms)were reported.The synthesis of magnetic iron oxide ultrafine particle and embolization effects of magnetic gelatin microspheres(MG-ms)in dog were studied.Adr- MG-ms consist of 2%(w/w)of adriamycin(Adr)as the core,and 68% of gelatin and 30% of magnetite as the shell with a mean particle size of 22 μm. In vitro experiment,the release rate of drug demonstrated that the microspheres have sustained-release properties.The average diameter of magnetic iron oxide was approximately l0 nm. Transcatheter embolization with MG-ms and ￣(99m)Tc-labelled MG-ms was performed under external magenet control in dog liver,respectively.Gamma photography and angiogram revealed that MG-ms level was almost equal both in left and right hepatic arteries without magnet,while with magnet(1200 Gs),MG-ms level in left hepatic artery(target site)was about 2.25 fold higher than in right hepatic artery,and few MG-ms in thyroid gland,brain and heart was observed.Results showed that the MG-ms is a promising embolic agent for treatment of hepatic cancer under external magnet control.

Reversal of Multidrug Resistance by Neferine in Adriamycin Resistant Human Breast Cancer Cell Line MCF-7/ADM

Objective: To study the reversal effect of neferine on adriamycin (ADM) resistant human breast cancer cell line MCF-7/ADM. Methods: The cytotoxic effect of Nef or ADM was determined by 3-[4, 5-dimethylthiazol-2.-yl], 5-diphenyl tetraxolium bromid (MTT) assay. Apoptosis and the expression of P-glycoprotein (P-gp) were detected by flow cytometry (FCM). The intracellular ADM concentration was measured by HPLC. Results: Nef at 1, 5, 10 mol/L decreased the IC50 of ADM to MCF-7/ADM from 11.63 g/mL to 4.59, 2.44, 0.27 g/mL respectively. MCF-7/ADM could resist the apoptosis induced by ADM while Nef (1-10 mol/L) could augment ADR-mediated apoptosis. Nef (10 mol/L) increased the accumulation of ADM up to 2.88 fold in MCF-7/ADM but not in sensitive cells MCF-7/S and reduced the expression of P-gp in MCF-7/ADM cells. Conclusion: Nef can circumvent multidrug resistance (MDR) of MCF-7/ADM cells and the mechanism was associated with the increase of intracellular accumulation of ADM and the reduced expression of P-gp in MCF-7/ADM cells.

A New Antifungal Flavonol Glycoside from Hypericum perforatum

In addition to six known flavonoids quercitrin, hyperoside, avicularin, rutin, quercetin and kaemferol, a new flavonol glycoside named 6″_O_acetyl quercetin 3_O_β_ D _alloside (1) was isolated from the aerial parts of Hypericum perforatum L. The structures were determined on the basis of spectroscopic methods (UV, IR, FAB_MS, 1H_NMR and 13 C NMR). Antifungal assay of all compounds showed that metabolite 1, quercitrin and quercetin were inhibitory to the growth of phytopathogenic fungus Helminthosporium sativum Pamel King et Bakke with minimum inhibitory concentrations (MICs) of 25, 50 and 50 μg/mL, respectively. Moreover, glycoside 1 and quercitrin were also shown to be able to inhibit the growth of Fusarium graminearum Schw. with MIC of 100 μg/mL. The MICs of ketoconazole used as control against the test fungi were 0.5 μg/mL in our assay.

Effect of Four Pesticides on Controlling Autumn Cowpea Greenhouse Whitefly

In order to mitigate the occurrence and damage of cowpea whitefly, reduce chemical pesticide consumption, residue and environmental pollution, the methods of forecasting, randomized block design andstatistical analysis were used in field efficacy trials, to study the control effect of cowpea anthracnose by usingchemical pesticides dinotefuran, acetamiprid, pymetrozine, and bio-pesticide avermectin. The results showed that after applying 1.85% avermectin EC 375 g, 20% dinotefuran SP 600 g, 5% acetamiprid WP 600 g, and 25% pymetrozine WP 600 g （control pesticide） per hm2 once every 5-8 days, and 3 times continuously underserious autumn occurrence of greenhouse whitefly, the control efficacy was 90.9%, 97.0%, 88.0%, 93.9%respectively on the 7th day after the pesticides application; 97.4%, 92.1%, 84.2%, 89.4% respectively on the 14th day after the pesticides application; and 95.6%, 86.7%, 82.2%, 84.5%, respectively on the 20th day afterthe pesticides application. The control effects among avermectin, acetamiprid and pymetrozine were not significantly different, while the effects of avermectin and pymetrozine were significantly higher than that of acetamiprid on the 7th day. The control effects among avermectin, dinotefuran and pymetrozine were not significantly different, while significantly higher than that of acetamiprid on the 14th day. The control effect of avermectin was significantly higher than that of dinotefuran, acetamiprid and pymetrozine on the 20th day. Meanwhile, the control effect of avermectin was slightly lower, higher and much higher than that of dinotefuran on the 7th, 14th, 20th day respectively after the treatment. These results indicated that the four pesticides,especiaLly biopesticide avermectin, were ideal pesticides, which could not only be used for controlling cowpea whitefly, but also realize modern, green, organic and ecological agricultural production.

Study on the Activity of Herbal Extract to Kill Root-knot Nematode of Cucumber Seedlings Stage

Present mature plants hydroponic technology was used,combined with some excellent characteristics,such as growth conditions was easy to control and process of root growth was easy to continuously observe,the nematicidal activity of 5 kinds of Chinese herbs extracts and the compound solution of Avermectin,with strong contact toxicity effect indoor,was systematically studied and investigated the affection on the root-knot nematode parasitized on the cucumber seeding stage. It is found that under the premise of no influence on root growth of cucumber,extracts from Picrorhiza scrophulariiflora and Punica granatum showed strong prevention and nematicidal activity,and had the similar efficacy of Avermectin; while the extracts from Cibotium barometz,Aucklandia lappa Decne and Fructus cnidii showed low nematicidal activity and various degrees inhibition effect on plant growth.

Validity and Systematic Position of Rana altaica(Rana:Ranidae):Results of a Phylogenetic Analysis

In order to evaluate the phylogenetic position and validity of Rana altaica,we investigated the phylogeny of brown frogs in Eurasia by Bayesian Inference and Maximum Parsimony analyses of a fragment from the mitochondrial DNA gene Cytochrome b.Both analyses resolved R.altaica as nesting deeply within R.arvalis.Most samples of the nominal R.altaica from the Altai region and specimens from Central Siberia shared a haplotype with R.arvalis based on the network analysis.The matrilineal relationships suggested that R.altaica should be considered as a junior synonym of R.arvalis.Furthermore,our study suggested that the species group division of Chinese brown frogs should be re-evaluated within a phylogenetic context.

Development of a New Azithromycin Glutamate for Parenteral Preparation, the Toxicity in Sprague-Dawley Rats and Pharmacokinetics in Human Healthy Volunteers

Aim In order to improve the solubility of azithromycin, the objectives of the present study were to screen an appropriate salt for azithromycin by comparing acute hepatic and renal toxicities in animals, and study the pharmacokinetics of final chosen azithromycin salt. Methods Various salts of azithromycin, such as glutamate, citrate, hydrochloride, sulphate, dihydrogen phosphate, lactobionate, tartrate, and aspartate were given intravenously to Sprague Dawley rats at a dose of 10 mg once daily for 14 consecutive days via tail vein. The acute hepatic and renal indicators were measured before and after administration. A pharmacokinetic study was performed on 12 healthy human volunteers. The subjects were equally divided into two groups by a randomized crossover design. Azithromycin glutamate injection was administered by intravenous infusion or intramuscular injection at a single dose of 500 mg, respectively. Azithromycin concentrations in plasma were determined by microbial inhibition zone assay, and the pharmacokinetic parameters were calculated using a practical pharmacokinetic software 3P87 program. Results Azithromycin glutamate was least toxic to the liver and kidney of the rats, thus being selected as a final salt for parenteral preparation of azithromycin. Pharmacokinetic results showed that the area under the plasma concentration-time curves （AUC0-120h） were 21.47 ± 1.57 h·μg·mL^-1 for intravenous infusion, and 19.36 ± 2.44 h·μg·mL^-1 for intramuscular injection. The absolute bioavailability of intramuscular injection was 92.59%. Conclusion Azithromycin glutamate is suitable for the future clinical application, and its pharmacokinetics is characterized in human volunteers in the present study.

Evolution of systemic therapy of advanced hepatocellular carcinoma

Hepatocellular carcinoma (HCC) commonly occurs in hepatitis B endemic areas, especially in Asian countries. HCC is highly refractory to cytotoxic chemotherapy. This resistance is partly related to its tumor biology, pharmacokinetic properties, and both intrinsic and acquired drug resistance. There is no convincing evidence thus far that systemic chemotherapy improves overall survival in advanced HCC patients. Other systemic approaches, such as hormonal therapy and immunotherapy, have also disappointing results. Recently, encouraging results have been shown in using sorafenib in the treatment of advanced HCC patients. In this review, we concisely summarize the evolution of developments in the systemic therapy of advanced HCC.

Synergistic growth inhibitory effects of Phyllanthus emblica and Terminalia bellerica extracts with conventional cytotoxic agents:Doxorubicin and cisplatin against human hepatocellular carcinoma and lung cancer cells

AIM: To examine the growth inhibitory effects of Phyllanthus emblica (P. emblica) and Terminalia bellerica (T. bellerica) extracts on human hepatocellular carcinoma (HepG2), and lung carcinoma (A549) cells and their synergistic effect with doxorubicin or cisplatin. METHODS: HepG2 and A549 cells were treated with P. emblica and T. bellerica extracts either alone or in combination with doxorubicin or cisplatin and effects on cell growth were determined using the sulforhodamine B (SRB) assay. The isobologram and combination index (CI) method of Chou-Talalay were used to evaluate interactions between plant extracts and drugs. RESULTS: P. emblica and T. bellerica extracts demonstrated growth inhibitory activity, with a certain degree of selectivity against the two cancer cell lines tested. Synergistic effects (CI < 1) for P. emblica /doxorubicin or cisplatin at different dose levels were demonstrated in A549 and HepG2 cells. The T. bellerica/ cisplatin or doxorubicin also showed synergistic effects in A549 and HepG2 cells. In some instances, the combinations resulted in antagonistic effects. The dose reduction level was different and specific to each combination and cell line. CONCLUSION: The growth inhibitory activity of doxorubicin or cisplatin, as a single agent, may be modified by combinations of P. emblica or T. bellerica extracts and be synergistically enhanced in some cases. Depending on the combination ratio, the doses for each drug for a given degree of effect in the combination may be reduced. The mechanisms involved in this interaction between chemotherapeutic drugs and plant extracts remain unclear and should be further evaluated.

Partial Beclin 1 silencing aggravates doxorubicin-and Fasinduced apoptosis in HepG2 cells

AIM： To investigate the role of Beclin 1 on the susceptibility of HepG2 cells to undergo apoptosis after anti-Fas antibody or doxorubicin treatment. METHODS： Beclin 1 silencing was achieved using RNA interference. DNA ploidy, the percentage of apoptotic cells and the mitochondrial membrane potential were assessed by flow cytometry. Levels of Beclin 1, BCI-XL and cytochrome c, and the cleavage of poly （ADP-ribose） polymerase （PARP） were assayed by using Western blots. RESULTS： Beclin 1 expression decreased by 75% 72 h after Beclin 1 siRNA transfection. Partial Beclin 1 silencing significantly increased the percentage of subG1 cells 24 and 40 h after treatment with doxorubicin or anti-Fas antibody, respectively, and this potentiation was abrogated by treatment with a pan-caspase inhibitor. Partial Beclin 1 silencing also increased PARP cleavage, mitochondrial membrane depolarization and cytosolic cytochrome c. The pro-apoptotic consequences of partial Beclin 1 silencing were not associated with a decline in Bcl-XL expression.CONCLUSION： Partial Beclin 1 silencing aggravates mitochondrial permeabilization and apoptosis in HepG2 cells treated with an anti-Fas antibody or with doxorubicin.

Effects of fructose-1,6-diphosphate on concentration of calcium and activities of sarcoplosnic Ca^(2+)-ATPase in cardiomyocytes of Adriamycin-treated rats

Objective: To observe the effects of fructose-1,6-diphosphate (FDP) on serum levels of cardiac troponin 1 (cTnl) and creatine kinase-MB (CK-MB), as well as the concentration of calcium in cardiomyocytes (Myo[Ca2+]) and activity of sarcoplosnic Ca2+-ATPase (SRCa2+-ATPase) in Adriamycin (ADR)-treated rats. Methods: Rats were intraperitoneally injected with ADR (2.5 mg/kg every other day for 6 times) and then with different dosages of FDP (every other day for twenty-one times). Bi-antibodies sandwich Enzyme linked immune absorption assay (ELISA) was performed to detect serum level of cTnl. CK-MB was detected by monoclonal antibody, Myo[Ca2+] was detected by fluorescent spectrophotometry and the activity of SRCa2+-ATPase was detected by inorganic phosphate method. Results: FDP (300, 600, 1200 mg/kg) significantly reduced the serum levels of cTnl and CK-MB, while at the same time decreased calcium concentration and increased SRCa2+-ATPase activity in cardiomyocytes of ADR-treated rats (P<0.01). Conclusions: FDP might alleviate the cardiotoxic effects induced by ADR through decreasing calcium level as well as increasing SRCa2+-ATPase activity in cardiomyocytes.

Treatment of hepatoma with liposome-encapsulated adriamycin administered into hepatic artery of rats

AIM： To observe the therapeutic effects of liposomeencapsulated adriamycin （LADM） on hepatoma in comparison with adriamycin solution （FADM） and adriarnycin plus blank liposome （ADM ＋ BL） administered into the hepatic artery of rats. METHODS： LADM was prepared by pH gradient-driven method. Normal saline, FADM （2 mg/kg）, ADM＋BL （2 mg/kg）, and LADM （2 mg/kg） were injected via the hepatic artery in rats bearing liver W256 carcinosarcoma, which were divided into four groups randomly. The therapeutic effects were evaluated in terms of survival time, tumor enlargement ratio, and tumor necrosis degree. The difference was determined with ANOVA and Dunnett test and log rank test. RESULTS： Compared to FADM or ADM ＋ BL, LADM produced a more significant tumor inhibition （tumor volume ratio： 1.243±0.523 vs 1.883±0.708, 1.847±0.661, P 〈 0.01）, and more extensive tumor necrosis. The increased life span was prolonged significantly in rats receiving LADM compared with FADM or ADM＋BL （231.48 vs 74.66, 94.70） （P 〈 0.05）. CONCLUSION： The anticancer efficacies of adriamycin on hepatoma can be strongly improved by liposomal encapsulation through hepatic arterial administration.

Protection of Calcitonin Gene-Related Peptide-Induced Preconditioning against Myocardial Injury due to Adriamycin in the Isolated Rat Heart *

The protective effect of calcitonin gene-related peptide (CGRP)-induced preconditioning on myocardial injury due to adriamycin was studied in the isolated perfused rat heart. Adriamycin (100 and 200 (mol / L) caused a gradual decrease in coronary flow (CF) and cardiac function (LVP and LV dp/dtmax), and an increase in the level of MDA. Pretreatment with CGRP at the concentration of 5 nmol/L for 5 min markedly reduced the attenuation of CF and cardiac function and inhibited the elevation of MDA content induced by adriamycin. The findings suggest that the pretreatment with CGRP possesses a protection against myocardial injury elicited by adriamycin. The present results also suggest that the protection of CGRP may be related to a reduction in lipid peroxidation.