中药治疗气分暑热型病毒性脑膜炎疗效分析

目的:观察分析中药治疗气分暑热型病毒性脑膜炎的疗效。方法:将82例患者随机分为对照组40例和研究组42例。两组均给予抗病毒、抗炎等基础治疗,研究组则额外予以中药颗粒剂白虎汤辨证加减治疗。比较两组退热和头痛消失时间、脑脊液7 d内恢复正常比例、C反应蛋白(CRP)水平及神经元特异性烯醇化酶(NSE)含量。结果:研究组退热时间、头痛消失时间短于对照组;脑脊液7 d内恢复正常比例高于对照组;CRP水平及NSE含量均低于对照组(P<0.05)。结论:中药辨证治疗气分暑热型病毒性脑膜炎有良效。

欧盟对生物杀灭剂有效成分评审后才会进行使用规范

在欧盟,对任何生物杀灭剂包括非农用农药可持续使用规范,都要等到对其已存在的有效成分评审完成后,才会出台。欧盟委员会环境总局的Pierre Choraine认为,在完成这项工作之前,市场上有很多产品依据现行欧盟标准难以登记,而且现在就进行使用规范的条件还不成熟。他是在九月份由欧洲化学品管理局(ECHA)组织的非农用农药业者日的活动上,发表上述观点的。Choraine先生提到。

欧盟对生物杀灭剂有效成分评审后才会进行使用规范

在欧盟,对任何生物杀灭剂包括非农用农药可持续使用规范,都要等到对其已存在的有效成分评审完成后,才会出台。欧盟委员会环境总局的Pierre Choraine认为,在完成这项工作之前,市场上有很多产品依据现行欧盟标准难以登记,而且现在就进行使用规范的条件还不成熟。他是在九月份由欧洲化学品管理局(ECHA)组织的非农用农药业者日的活动上,发表上述观点的。Choraine先生提到。

Pollution of Environmental Endocrine Disrupting Chemicals(EDCs) in Water and Its Adverse Reproductive Effect on Fish

Environmental endocrine disrupting chemicals （EDCs）, commonly found in the environment, come from industry and agriculture, including pesticides, fungicides, insecticides, herbicides, and other chemicals. Nowadays, more and more EDCs were released into the environment. EDCs go into water body via atmosphere sedi-mentation, surface runoff, soil eluviation, etc., so water body becomes the main place for existing. In order to attract scientific and public attention worldwide and to prevent EDCs pol ution, in this study we reviewed the classification of EDCs and their concentrations in natural water bodies, drinking water sources and water plants, and the reproductive toxicity of EDCs to fish were reviewed. EDCs could disturb the endocrine system and make reproductive organs and reproduction abnor-mal, resulting in fertility descending, reproduction function damage, community quan-tity decrease and even species extinction. In addition, EDCs could disrupt the homeostasis maintained by hormones, which would result in defects of neural de-velopment and abnormalities of the endocrine and reproductive systems. The exact molecular mechanisms have not been completely reported, but researches have suggested that multiple mechanisms were involved in the action of EDCs. Although there have been researches on the biohazard of EDCs, there stil exist problems of weakness in fundamental researches, difficulties in recognizing and identifying EDCs and high cost, which restraint the knowledge on them.

Desiccation Alters the Stability and Distribution of Pea Seed_borne Mosaic Virus in Pea (Pisum sativum) Cotyledon Cells

As a seed transmitted pathogen, pea seed_borne mosaic virus (PSbMV) not only replicates in embryonic cells but can also withstand seed desiccation. To understand the mechanism of PSbMV tolerance to seed desiccation, the authors compared the stability of viral coat protein (CP) and the distribution of viral particles in the cotyledon cells of pea (Pisum sativum L.) embryos collected before and after the dehydration process. Before dehydration, when the embryo was fresh and immature, degradation of CP was observed and a predominantly perinuclear distribution of viral particles in the cotyledon cells was evident. After dehydration, when the embryo was dry and mature, degradation of CP did not occur and the perinuclear viral distribution disappeared. Instead, aggregates containing PSbMV CP were found in the cytoplasm. Electron microscopy showed that these aggregates were composed of PSbMV particles. The formation of PSbMV particle aggregates is apparently triggered by seed dehydration and may be favorable to the virus survival in the desiccated embryonic cells.

Constituents from Ranunculus sieboldii Miq.

Aim To investigate the chemical composition of Ranunculus sieboldii Miq..Methods Repeated column chromatography over silica gel, polyamide and RP-18 followed by gelfiltration on sephadex LH-20 were used to isolate chemical constituents, and their structures wereelucidated by extensive spectroscopic methods (UV, IR, MS, ~1H NMR, ^(13)C NMR) including 2D NMR(COSY, HMQC, HMBC, NOESY) techniques and by direct comparing spectral data with those reported inliterature. Results Five flavonoid glycosides named apigenin-4'-O-α-L-rhamnopyranoside (1),apigenin-7-O-β-D-glucopyranosyl-4'-O-α-L-rhamnopyranoside (2), apigenin-8-C-α-L-arabinopyranoside(3), apigenin-8-C-β-D-ga-lactopyranoside (4) , tricin-7-O-β-D-glucopyranoside (5), together withtricin (6), luteolin (7), scopoletin (8), esculetin (9), scoparone (10), ferulic acid (11),protocatechuic acid (12) , and tematolide (13) were isolated from the 95% etha-nolic extract of itswhole plant, and their cytotoxic activities were preliminarily tested. Conclusion Compounds 1-12were obtained from this genus and compound 13 from this species for the first time. Furthermore,compound 1 was for the first time isolated from nature while the ^(13)C NMR data of compounds 2 and3 are reported for the first time. The bioassay revealed that compound 1 was active against BEL-7407and A549 cell lines (IC_(45) 43, 77 μg·mL^(-1)), 8 and 10 showed inhibitory activities on KB celllines (IC_(50) 78, 44 μg·mL^(-1)) and HL-60 cell lines (Ic_(50) 85, 85 μg·mL^(-1)), while 7exerted moderate cytotoxic activities on KB, BFL-7407, A549 and HL-60 cell lines with their IC_(50)being 51, 55, 44 and 10 μg·mL^(-1) , respectively.

Expression of Porcine Reproductive and Respiratory Syndrome Virus ORF7 Gene and Purification and Immunological Activity Analysis of the Recombinant Protein

[Objective] The aim of this study was to realize efficient expression of the porcine reproductive and respiratory syndrome virus （PRRSV） ORF7 gene in genetic engineering bacteria and analYze the immunological activity of the recombinant protein after purification. [ Method] The constructed recombinant expression vector pET-ORF7 was transformed into Escherichia co1BL21 （DE3） and induced by IPTG under the optimal condition. After analysis of SDS-PAGE and Western Blot, the expression products were purified by Ni-NTA His · Bind Resin chrom- atographic column under denaturing condition and renatured by gradient dialysis. Subsequently, the immunological activity of the renatured recombinant protein was detected by Westem Blot and indirect ELISA. [ Result] The recombinant plasmid pET-ORF7 expressed in E. coli successfully, and the fusion protein was in the form of inclusion body. By SDS-PAGE detection, the molecular weight of the expression protein was approximate 33 kD, according with the expectation. Analysis by Bandscan software showed that the expressed fusion protein was about 50% of total bacterial protein of BL21 （DE3）. Wastem Blot and indirect ELISA detection showed that the renatured protein could react with PRRSV positive serum specifically, indicating its good immunological activity. [ Conclusion] This study lays a foundation for the preparation of PRRSV monoclonal antibody and diagnostic kit.

Comparative Study of the Biological Activities of the Skin Secretions from Six Common Chinese Amphibians

Water soluble skin secretions of six common Chinese amphibians were studied for their biological and enzymatic activities.The skin secretions of Tylototriton verrucosus,Bombina maxima ,and Bufo andrewsi were found toxic to mice with the intraperitoneal LD 50 of 11 5?mg/kg,18 8?mg/kg,and 264?mg/kg,respectively.No acute lethal toxicities were observed for the skin secretions of Rana nigromaculata,Rana guentheri and Rana limnocharis in a dose up to 500?mg/kg.The lethal toxicities of the skin secretions of T verrucosus and B maxima to mice are in the same grade as those of Viperidae snake venoms.The toxic components in T verrucosus and B maxima skin secretions are the proteins with molecular weights ranging from 3 to 60?kDa.All the skin secretions had both proteolytic activity and trypsin inhibitory activity.The skin secretions from T verrucosus , B maxima and B andrewsi also displayed wide spectrum antimicrobial activity.On the other hand,the skin secretions from B andrewsi and B maxima showed cytotoxicity on human cancer cells.All the six samples had not significant effects on mammalian blood coagulation system.Phospholipase A 2 activity was only found in the skin secretions of T verrucosus .None of these skin secretions showed acetylcholine esterase activity.

Radiation therapy concurrent with weekly cisplatin therapy for loco-regionally advanced nasopharyngeal carcinoma:outcomes of a clinical trial

Objective:The purpose of this study was to define the maximum tolerated dose(MTD) by describing the doselimiting toxicity(DLT) of weekly cisplatin concurrently with conventional plus 3-dimensional conformal radiotherapy(CT + 3DCRT) in patients with loco-regionally advanced nasopharyngeal carcinoma(NPC).Methods:Patients with loco-regionally advanced NPC(III/IVa stage) were enrolled into a dose-escalating study.Toxicity was graded according to Common Terminology Criteria for Adverse Events version 3.0(CTCAE v3.0).MTD was defined when 2 of 6 patients developed DLT.The starting dose of cisplatin was 15 mg/m2/w,with a subsequent dose escalation of 5 mg/m2/w in cohorts of 3 new patients.CT + 3DCRT was given to the nasopharynx and the upper neck;the lower neck was treated by a single anterior field irradiation.The prescription dose was 70-80 Gy by 35-40 fractions to the nasopharynx gross tumor,and 66-70 Gy by 33-35 fractions to the positive neck lymph nodes.Results:From Jun.2008 to Sep.2009,24 patients received complete chemoradiotherapy,and all of them were eligible for toxicity evaluation.On the first five dose levels of 15 mg/m2/w and 35 mg/m2/w,no patient experienced DLT.On the next dose level of 40 mg/m2/w,1 patient experienced DLT of grade 3 myelosuppression for 1.4 weeks,and among the additional 3 patients,no one developed DLT.On the seventh dose level of 45 mg/m2/w,all the patients developed grade 3 myelosuppression for more than 1 weeks,and the dose-escalating trial stopped.The 23(95.8%) patients achieved clinical complete remission(CR) in the local site;22(91.7%) achieved CR in the regional site,and all patients got CR 3 months later.After a median follow-up of 16.4 months,1 patient developed liver metastases 2 months later,1 patient developed bone metastases 10 months later and 22 kept disease-free survival.Conclusion:The MTD of cisplatin weekly with concurrent CT + 3DCRT for local advanced NPC is 40 mg/m2/w,with myelosuppression as DLT.

Indinavir Resistance Evolution in One Human Immunodeficiency Virus Type 1 Infected Patient Revealed by Single-Genome Amplification

Human Immunodeficiency Virus Type 1 exists in vivo as quasispecies, and one of the genome's characteristics is its diversity. During the antiretroviral therapy, drug resistance is the main obstacle to effective viral prevention. Understanding the molecular evolution process is fundamental to analyze the mechanism of drug resistance and develop a strategy to minimize resistance. Objective: The molecular evolution of drug resistance of one patient who had received reverse transcriptase inhibitors for a long time and had treatment which replaced Nevirapine with Indinavir was analyzed, with the aim of observing the drug resistance evolution pathway. Methods: The patient, XLF, was followed-up for six successive times. The viral populations were amplified and sequenced by single-genome amplification. All the sequences were submitted to the Stanford HIV Drug Resistance Database for the analysis of genotypic drug resistance. Results: 149 entire protease and 171 entire reverse transcriptase sequences were obtained from these samples, and all sequences were identified as subtype B. Before the patient received Indinavir, the viral population only had some polymorphisms in the protease sequences. After the patient began Indinavir treatment, the variants carrying polymorphisms declined while variants carrying the secondary mutation G73S gained the advantage. As therapy was prolonged, G73S was combined with M46I/L90M to form a resistance pattern M46I/G73S/L90M, which then became the dominant population. 97.9% of variants had the M46I/G73S/L90M pattern at XLF6. During the emergence of protease inhibitors resistance, reverse transcriptase inhibitors resistance maintained high levels. Conclusion: Indinavirresistance evolution was observed by single-genome amplification. During the course of changing the regimen to incorporate Indinavir, the G73S mutation occurred and was combined with M46I/L90M.

Aptamers Against Viral Hepatitis:from Rational Design to Practical Application

Aptamers are short nucleic acids or peptides that strongly bind to a protein of interest and functionally inhibit a given target protein at the intracellular level. Besides high affinity and specificity, aptamers have several advantages over traditional antibodies. Hence, they have been broadly selected to develop antiviral agents for therapeutic applications against hepatitis B and C viruses (HBV, HCV). This review provides a summary of in vitro selection and characterization of aptamers against viral hepatitis, which is of practical significance in drug discovery.

Neither gastric topological distribution nor principle virulence genes of Helicobacter pylori contributes to clinical outcomes

AIM: Studies on Helicobacter pylori (H pylon) and gastroduodenal diseases have focused mainly on the distal sites of the stomach, but relationship with the gastric cardia is lacking. The aim of this study is to determine if the gastric topology and genotypic distribution of Hpyloriwere associated with different upper gastrointestinal pathologies in a multiethnic Asian population. METHODS: Gastric biopsies from the cardia, body/corpus and antrum were endoscoped from a total of 155 patients with dyspepsia and/or reflux symptoms, with informed consent. H pylori isolates obtained were tested for the presence of 26kDa, ureC, cagA, vacA, iceA1, iceA2 and babA2 genes using PCR while DNA fingerprints were generated using random amplification polymorphic DNA (RAPD). RESULTS: Hpyloriwas present in 51/155 (33%) of patients studied. Of these, 16, 15 and 20 were isolated from patients with peptic ulcer diseases, gastroesophageal reflux diseases and non-ulcer dyspepsia, respectively. Of the Hpyloripositive patients, 75% (38/51) had Hpyloriin all three gastric sites. The prevalence of various genes in the H pylori isolates was shown to be similar irrespective of their colonization sites as well as among the same site of different patients. The RAPD profiles of H pylori isolates from different gastric sites were highly similar among intra-patients but varied greatly between different patients. CONCLUSION: Topographic colonization of H pylori and the virulence genes harboured by these isolates have no direct bearing to the clinical state of the patients. In multiethnic Singapore, the stomach of each patient is colonized by a predominant strain of H pylori，irrespective of the clinical diagnosis.

Undifferentiated embryonal sarcoma of liver in an old female:Case report and review of the literature

The clinical characteristics of undifferentiated(embryonal) sarcoma of the liver(UESL) were investigated and the best treatment modalities were recommended.Both histology and immuno-histochemistry demonstrated the cellular features of this peculiar tumor.The tumor size was 12 cm × 9 cm × 8 cm in the right liver lobe.The patient underwent surgical resection of the tumor.The postoperative recovery was uneventful and she died eight months after diagnosis.The tumor showed mixed spindle and polygonal cells within the myxoid matrix.Some tumor cells contained eosinophilic hyaline globules that were positive for resistant diastase.Immunohistochemistry showed positive vimentin.Stellate and spindle cells were positively stained with alpha-1-antichymotrypsin(AACT) and CD68.This case indicates that UESL is not obviously differentiated in old-aged adults.

Silencing of UBP43 by shRNA Enhances the Antiviral Activity of Interferon against Hepatitis B Virus

Previous studies have shown that expression of the interferon-sensitive gene （ISG）15 protease UBP43 is increased in the liver biopsy specimens of patients who do not respond to interferon （IFN）-α therapy. We hypothesized that UBP43 might hinder the ability of IFN to inhibit HBV replication. In this study, we investigated whether vector-based siRNA promoted by HI （psiUBP43） could enhance IFN inhibiting HBV replication in cell culture. UBP43 was specifically silenced using shRNA. In HepG2.2.15 cells, the HBeAg and HBV DNA levels were significantly reduced by IFN after transfection of shRNA, imphicated that vector-based siRNA promoted by H1 （psiUBP43） could enhance IFN inhibiting HBV replication in cell culture. These data suggest that UBP43 modulates the anti-HBV type I IFN response, and is a possible therapeutic target for the treatment of HBV infection.

Genetic Characteristics of 2009 Pandemic H1N1 Influenza A Viruses Isolated from China's Mainland

A total of 100 H1N1 flu real-time-PCR positive throat swabs collected from fever patients in Zhejiang, Hubei and Guangdong between June and November 2009, were provided by local CDC laboratories. After MDCK cell culture, 57 Influenza A Pandemic （H1N1） viruses were isolated and submitted for whole genome sequencing. A total of 39 HA sequences, 52 NA sequences, 36 PB2 sequences, 31 PB1 sequences, 40 PA sequences, 48 NP sequences, 51 MP sequences and 36 NS sequences were obtained, including 20 whole genome sequences. Sequence comparison revealed they shared a high degree of homology （96%-99%） with known epidemic strains （A/Califomia/04/2009（H1N1）. Phylogenetic analysis showed that although the sequences were highly conserved, they clustered into a small number of groups with only a few distinct strains. Site analysis revealed three substitutions at loop 220 （221-228） of the HA receptor binding site in the 39 HA sequences： A/Hubei/86/2009 PKVRDQEG→PKVRDQEA, A/Zhejiang/08/2009 PKVRDQEG→PKVRDQER, A/Hubei/75/2009 PKVRDQEG→PKVRDQGG, the A/Hubei/75/2009 was isolated from an acute case, while the other two were from patients with mild symptoms. Other key sites such as 119, 274, 292 and 294 amino acids of NA protein,627 of PB2 protein were conserved. Meanwhile, all the M2 protein sequences possessed the Ser32Asn mutation, suggesting that these viruses were resistant to adamantanes. Comparison of these sequences with other H1N1 viruses collected from the NCBI database provides insight into H1N1 transmission and circulation patterns.

Estimation and Test of Interaction Parameters in the UNIFAC Model of Group-OCOO-with Group-CH_3,ACH, CH_3OH and CH_3COO-

Isobaric vapor-liquid equilibria (VLE) are experimentally measured for the binary systems of dimethyl carbonate (DMC)+ ethylene carbonate and methanol + ethylene carbonate at 101.325kPa. The thermodynamic consistency of these experimental data is tested with an available statistic method. Interaction parameters of the carbonate group -OCOO- with the group -CH3, ACH, CH3OH and CH3COO- in UNIFAC model are determined using the experimental and literature VLE data. The results show that the calculated VLE data using the new UNIFAC parameters agree excellently with the experimental data in this work and in literature. These results are useful in the research on DMC and diphenyl carbonate synthesis by transesterification in design of reactor and distillation tower.

Circulating adhesion molecules in patients with virus-related chronic diseases of the liver

AIM: In the inflammatory state, intercellular adhesion molecule-1 (ICAM-1) and vascular cellular adhesion molecule-1 (VCAM-1) play a key role in promoting migration of immunological cells from the circulation to target site.Aim of our study was to investigate soluble forms of these molecules in patients with virus-related chronic liver diseases, to assess their behavior in different pathologies and correlation with severity of liver damage.METHODS: Circulating ICAM-1 and VCAM-1 were assayed by EIA commercial kits (R&D System Co.,Abington, UK) in 23 patients with chronic active hepatitis (CH), 50 subjects affected by liver cirrhosis (LC) and 15 healthy controls comparable for sex and age. In patients, serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were also detected by autoanalyzer.RESULTS: LC patients had significantly higher ICAM-1 values than CH patients (38.56±7.4 ng/mL vs 20.89±6.42 ng/mL; P＜0.001) and these ones had significantly higher values than controls (12.92±1.08 ng/mL; P＜0.001). In CH group, ICAM-1 levels were significantly related to inflammatory activity (P= 0.041) and ALT values (r= 0.77;P＜0.05). VCAM-1 values were significantly increased only in LC patients (P＜0.001) and related to severity of liver impairment.CONCLUSION: These findings suggest that the determination of serum ICAM-1 can be considered as an additional useful marker of hepatocellular necrosis and inflammatory activity in chronic hepatitis, while serum VCAM-1 is an indicator of liver fibrogenesis and severity of disease in cirrhosis.

Molecular Characterization of Nonstructural Protein NS38 of Grass Carp Reovirus

Viral nonstructural proteins in both enveloped and non-enveloped viruses play important roles in viral replication. Protein NS38 of Grass carp reovirus (GCRV), has been deduced to be a non-structural protein, and, consistent with other reoviruses, is considered to cooperate with the NS80 protein in viral particle assembly. To investigate the molecular basis of the role of NS38, a complete protein was expressed in E.coli for the first time. It was found that there is a better expression of NS38 induced with IPTG at 28 ℃ rather than 37 ℃. In addition, the antiserum of NS38 prepared with purified fusion protein and injected into rabbit could be used for detecting NS38 protein expression in GCRV infected cell lysate, while there is not any reaction crossed with purified virus particle, confirming NS38 is not a component of the viral structural protein. The result reported in this study will provide evidence for further viral protein-protein and protein-RNA interaction in dsRNA viruses replication.

Isolation, identification, and cytotoxicity of a new isobenzofuran derivative from marine Streptomyces sp. W007

A new isobenzofuran derivative( 1) was isolated from the marine Streptomyces sp. W007 and its structure was determined through extensive spectroscopic analyses, including 1D-NMR, 2D-NMR, and ESI-MS. The absolute configuration of compound 1 was determined by a combination of experimental analyses and comparison with reported data, including biogenetic reasoning, J-coupling analysis, NOESY, and 1 H- 1 HCOSY. Compound 1 exhibited no cytotoxicity against human cells of gastric cancer BGC-823, lung cancer A549, and breast cancer MCF7.

Toxicity and Accumulation of Selenite in Four Microalgae

The toxicity and bioaccumulation of selenite in four microalgae, Spirulina platensis, Dunaliella salina, Dunaliella bardawill and Phaeodactylum tricornutum cultured in the presence of selenite were investigated. Lower concentrations of selenite were generally nontoxic and frequently stimulated algal growth, while higher concentrations of selenite inhibited algal growth. Selenite was more toxic to D. salina and D. bardawill than to S. platensis and P. tricornutum . All algae cultured in selenite were able to incorporate Se to different degrees, which depended on algal species. The distributions of selenite among intracellular macromolecular compounds were different among algal species: most of the selenite was associated with proteins in S. platensis, D. salina and D. bardawill , while most of the selenite was associated with lipids in P. tricornutum , which reflected the physiological differences among the algae. These observations suggest that algae are able to accumulate selenite and bind it with intracellular macromolecular compounds when exposed to high concentration of selenite. This may represent a form of storage or detoxification of selenite by the algae.