薄层扫描法测定抗心毒颗粒中黄芪甲苷的含量

目的 :建立用薄层扫描法测定抗心毒颗粒中黄芪甲苷含量的方法。方法 :甲醇提取 ,乙醚脱脂 ,正丁醇萃取后蒸干 ,上D10 1型大孔树脂柱洗脱 ;以 [正丁醇 -醋酸乙酯 -水 (4∶1∶5 )的上层液 ]-甲醇 (10∶1)为展开剂 ,置氨蒸汽饱和的展开缸内 ,喷以 10 %硫酸乙醇溶液 ,10 0℃至斑点显色。检测波长 :λs =5 30nm ,λr =70 0nm ,狭缝 2× 1.2 ,线性参数Sx=3。结果 :3批样品中黄芪甲苷的平均含量为 0 .5 2 0mg/袋 ,平均回收率为 96 .90 %。结论 :上述方法简单、准确 。

A New Technique for Whole Clearing

[Objective] The research aimed to study a new whole clearing and preparation technology.[Method] After young seeds of Stephania sp.preserved in the alcohol solution,as well as the fresh pistil and nucellus of Sorghum bicolor were separated,they were soaked and cleared in the solution of 84 disinfectant.And the plant materials could be mounted by using three methods.[Result] The globular embryos and heart-shaped embryos at different stages of Stephania sp.as well as the pistil,nucellus,embryo sac,hypostase o...

生命倒计时

据说,非洲有个民族,婴儿呱呱坠地就算是有60岁的寿命,以后逐年递减,直到零岁。人生大事都是在这60年内内完成,此后便是颐养天年的岁月了。 这算得上是个绝妙智慧恩德计岁方法。它把平常做惯了的“加法”变成做“减法”,为自己设定了一个生命的“倒计时”。倒计时原本是对某个特定时间的警示。

春季需排毒

从中医角度看，冬季具有封藏、收引的特性。在捂了一个漫长的冬季后，人体的脏腑内很容易堆积各种毒素，所以春季需要及时为脏腑排毒。

每当我把《冶金财会》翻开……

每当我把《冶金财会》翻开, 我仿佛遨游在知识的浩翰大海。 喜读那满刊百花齐放丰富多彩的文章, 使我大开眼界从中受益匪浅! 翻阅那来自四面八方的经济信息, 喜看冶金工业蓬勃发展捷报频传, 面对改革开放的祖国日新月异, 真是令我欢欣鼓舞心喜满怀。

《金瓶梅》中妇女形象初探

探析小说反映明代丑恶社会本质深度腐蚀当时妇女思想品性 ,以妇女容态言谈描写塑造妇女贪财、附势、淫荡、暴唳人物形象 ,研究小说塑造与之对立端庄、正直、贤慧、善良妇女形象 ,认识小说描写各异妇女形象之社会现实意义、挖掘小说所刻划妇女形象值得批判、借鉴、肯定、扬弃重要艺术价值。

Detection of hepatitis C virus core antigen for early diagnosis of hepatitis C virus infection in plasma donor in China

AIM： To evaluate the efficacy of a new hepatitis C virus （HCV） core antigen assay developed in China. METHODS： After the determination of HCV infection, 49 serial samples were selected from II regular plasma donors in 5 different plasma stations. To compare the performance of HCV core antigen detection and HCV PCR, these samples were genotyped, and each specimen was analyzed by ELISA for the detection of HCV core antigen and by qualitative HCV PCR. RESULTS： Among all of the sequential samples, the original 23 specimens were HCV RNA-negative, and 36 samples were HCV RNA-positive. Twenty-seven samples （75%） were HCV core antigen-positive from these HCV RNA-positive specimens. Conversely, 27 samples （93.2%） were found HCV RNA-positive in HCV core antigen- positive samples. Intervals between HCV RNA and HCV core antigen-positive, as well as between HCV core antigen-positive and HCV antibody-positive were 36.0 and 32.8 d, respectively. CONCLUSION： This HCV core antigen assay, developed in China, is able to detect much of anti-HCV-negative, HCV RNA-positive preseroconversion window period （PWP） plasma donations.

Analysis of Cardiotoxicity from rh-Endostatin Therapy Combined with Chemotherapy

OBJECTIVE To evaluate the cardotoxicity from recombinant human endostatin(rh-endostatin)combined with chemotherapy. METHODS A total of 12 cancer patients treated with rh- endostatin combined with chemotherapy were selected,and their clinical data collected.Their symptoms,including cardiopalmus, chest distress,dyspnea and changes in their electrocardiogram (ECG),myocardium enzymogram and left ventricular ejection fraction(LVEF),were observed during the drug treatment.These indicators were used for early diagnosis of cardiotoxicity. RESULTS Compared with a pre-therapeutic value,there was a significant increase in the CK-MB value at one week after starting the treatment as well as at the end of treatment(P<0.05).There was a significant change in the ECG at the end of treatment, compared to a pre-therapeutic condition(P<0.05),but there was no significant difference when comparing the pre-and post- therapeutic LVEF values. CONCLUSION It was recognized that mild cardiac adverse reactions exist in the regimen of recombinant human endostatin combined with chemotherapy.This therapy caused definite injury to the cardiac muscle,but cardiac functions were not obviously changed.CK-MB and ECG may be used as indicators for early monitoring cardiac toxicity.Vigilance against cardiac adverse reactions should be heightened during a course of rh-endostatin combined with chemotherapy.

Recombinant adeno-associated virus serotype 9 with p65 ribozyme protects H9c2 cells from oxidative stress through inhibiting NF-κB signaling pathway

Background Oxidative stress is a major mechanism underlying the pathogenesis of cardiovascular disease. It can trigger inflammatory cascades which are primarily mediated via nuclear factor-κB （NF-κB）. The NF-κB transcription factor family includes several subunits （p50, p52, p65, c-Rel, and Rel B） that respond to myocardial ischemia. It has been proved that persistent myocyte NF-κB p65 activation in heart failure exacerbates cardiac remodeling. Mechods A recombinant adeno-associated virus serotype 9 carrying enhanced green fluorescent protein and anti-NF-κB p65 ribozyme （AAV9-R65-CMV-eGFP） was constructed. The cells were assessed by MTT assay, Annexin V–propidium iodide dual staining to study apoptosis. The expression of P65 and P50 were assessed by Western blot to investigate the under-lying molecular mechanisms. Results After stimulation with H2O2 for 6 h, H9c2 cells viability decreased significantly, a large fraction of cells underwent apoptosis. We observed a rescue of H9c2 cells from H2O2-induced apoptosis in pretreatment with AAV9-R65-CMV-eGFP. Moreover, AAV9-R65-CMV-eGFP decreased H2O2-induced P65 expression. Conclusions AAV9-R65-CMV-eGFP protects H9c2 cells from oxidative stress induced apoptosis through down-regulation of P65 expression. These observations indicate that AAV9-R65-CMV-eGFP has the potential to exert cardioprotective effects against oxidative stress, which might be of great importance to clinical efficacy for cardiovascular disease.

Genes transactivated by hepatitis C virus core protein, a microarray assay

AIM: To explore the new target genes transactivated by hepatitis C virus (HCV) core protein and to elucidate the pathogenesis of HCV infection. METHODS: Reverse transcribed cDNA was subjected to microarray assay. The coding gene transactivated by HCV core protein was cloned and analyzed with bioinformatics methods. RESULTS: The expressive vector of pcDNA3.1(-)-core was constructed and confirmed by restriction enzyme digestion and DNA sequencing and approved correct. mRNA was purified from HepGZ and HepG2 cells transfected with pcDNA3.1(-)-core, respectively. The cDNA derived was subjected to microarray assay. A new gene named HCTP4 was cloned with molecular biological method in combination with bioinformatics method. CONCLUSION: HCV core is a potential transactivator. Microarray is an efficient and convenient method for analysis of differentially expressed genes.

Enhancement of CTLs induced by DCs loaded with ubiquitinated hepatitis B virus core antigen

AIM： To investigate whether hepatitis B virus （HBV） could induce a hepatitis B virus core antigen （HBcAg）specific cytotoxic T lymphocyte （CTL） response in vitro by dendritic cells （DCs） transduced with lentiviral vector-encoding ubiquitinated hepatitis B virus core antigen （LV-Ub-HBcAg）.METHODS： Recombinant LV-Ub-HBcAg were transfected into highly susceptible 293 T cells to obtain high virus titres, Bone marrow-derived DCs isolated from BALB/c mice were cultured with recombinant granulocyte-macrophage colony-stimulating factor and recombinant interleukin （IL）-4. LV-Ub-HBcAg, lentiviral vector-encoding hepatitis B virus core antigen （LV-HBcAg）, lentiviral vector （LV） or lipopolysaccharide were added to induce DC maturation, and the DC phenotypes were analyzed by flow cytometry. The level of IL-12 in the supernatant was detected by enzyme-linked immunosorbent assay （ELISA）. T lymphocytes were proliferated using Cell Counting Kit-8. DCs were cultured and induced to mature using different LVs, and co-cultured with allogeneic T cells to detect the secretion levels of IL-2, IL-4, IL-10and interferon-γ in the supernatants of T cells by ELISA. Intracellular cytokines of proliferative T cells were analyzed by flow cytometry, and specific CTL activity was measured by a lactate dehydrogenase release assay.RESULTS： LV-Ub-HBcAg-induced DCs secreted more IL-12 and upregulated the expression of CD80, CD86 and major histocompatibility class ]I, DCs sensitised by different LVs effectively promoted cytokine secretion; the levels of IL-2 and interferon-y induced by LV-Ub- HBcAg were higher than those induced by LV-HBcAg, Compared with LV-HBcAg-transduced DCs, LV-Ub- HBcAg-transduced DCs more efficiently stimulated the proliferation of T lymphocytes and generated HBcAgspecific cytotoxic T lymphocytes.CONCLUSION： LV-Ub-HBcAg effectively induced DC maturation. The mature DCs efficiently induced T cell polarisation to Thl and generated HBcAg-specific CTLs.

Mutations in hepatitis B virus core regions correlate with hepatocellular injury in Chinese patients with chronic hepatitis B

AIM: To elucidate the relationship between the frequency of core mutations and the clinical activity of hepatitis B virus (HBV)-related liver disease and to characterize the amino acid changes in the core region of HBV.METHODS: We studied 17 Chinese patients with chronic hepatitis B according to their clinical courses and patterns of the entire core region of HBV.RESULTS: Amino acid changes often appeared in the HBV core region of the HBV gene in patients with high values of alanine aminotransferase (ALT) or with the seroconversion from HbeAg to anti-HBe. The HBV core region with amino acid changes had high frequency sites that corresponded to HLA Ⅰ/Ⅱ restricted recognition epitopes reported by some investigators.CONCLUSION: The core amino acid changes of this study occur due to influence of host immune system. The presence of mutations in the HBV core region seems to be important for predicting the clinical activity of hepatitis B in Chinese patients.

THE COEXPRESSION OF THE preS1(1-42) AND THE CORE(1-144)ANTIGEN OF HBV IN E.coli

Objective.To study the therapeutic T cell vaccine for the treatment of chronic hepatitis B by improving the cellular immunization of HBsAg vaccine with the coexpression of the preS1(1-42)and the Core(1-144)anti-gen of HBV in E.coli.Methods.The genes of HBcAg (1-144)and preS1(1-42)were amplified and fused by PCR.This fused gene was inserted in the prokaryotic expression vector pET-11d and expre ssed in E.coli.Results.It was showed by SDS-PAGE that the pr otein molecular weight of the coexpr ession product was about 20kD,20%of all bacteria prote in.The monoclonal antibodies again st core and preS1antibody could re-act with this fused protein by Western-blot technique respectively.The fused gene was verified by sequencin g.Under the immune electron microscop y,this fused protein is typical particles of HBcAg but in an aggregated form.Conclusion.The results might aid for studying T c ell immunotherapeutic vaccine for c hronic hepatitis B.

Biomarkers with Potential Predictive Value for Cardiotoxicity in Anticancer Treatments

Rapid development of anticancer treatments in recent years has greatly improved prognosis of cancer patients.However,with extension of survival time of cancer patients,various short-term and long-term side effects brought about by anticancer treatments,especially cardiotoxicity,have become increasingly prominent.Nonetheless,at present,there is few diagnostic methods with extremely high sensitivity and specificity to detect and accurately predict whether patients with anticancer treatment will experience cardiovascular complications.Inflammation,fibrosis and oxidative stress are considered to be important mechanisms involved in cardiotoxicity anticancer treatments.The cardiovascular biomarkers having the ability to predict and detect cardiovascular dysfunction earlier than clinical symptoms as well as left ventricular ejection fraction monitored by echocardiography,are of great value to timely treatment adjustment and prognosis evaluation.Cardiac troponin T/I and brain natriuretic peptide/N-terminal prohormone of brain natriuretic peptide have been routinely used in clinical practice to monitor cardiotoxicity,and some new biomarkers such as soluble suppression of tumorigenecity-2,myeloperoxidase,growth differentiation factor-15,galectin-3,endothelin-1,have potential in this area.In the future,larger-scale experimental studies are needed to provide sufficient evidences,and how to detect them quickly and at low cost is also a problem to be dealed with.

Psychological impact of chronic hepatitis C:Comparison with other stressful life events and chronic diseases

AIM： To examine the psychological impact of chronic hepatitis C （CHC） diagnosis in a large cohort of CHC patients as compared with other stressful life events and chronic diseases carrying a risk of life-threatening complications. METHODS： One hundred and eighty-five outpatients with compensated CHC were asked to self-grade, using a 100-mm visual analogue scale （VAS）, the degree of stress caused by the learning of CHC diagnosis and the perceived severity of their disease. Diagnosis-related stress was compared to four other stressful life events and perceived CHC severity was compared to four other common chronic diseases. RESULTS： Learning of CHC diagnosis was considered a major stressful event （mean ± SD scores： 72±25）, significantly less than death of a loved-one （89±13, P〈0.0001） and divorce （78 ± 23, P〈0.007）, but more than job dismissal （68 ± 30, P〈 0.04） and home removal （26±24, P〈 0.0001）. CHC was considered a severe disease （74± 19）, after AIDS （94±08, P〈 0.001） and cancer （91± 11, P〈 0.001）, but before diabetes （66±23, P〈0.001） and hypertension （62±20, P〈0.001）. Perceived CHC severity was not related to the actual severity of liver disease, assessed according to Metavir fibrosis score. In multivariate analysis, diagnosisrelated stress was related to perceived disease severity （P〈0.001）, trait anxiety （P〈 0.001） and infection through blood transfusion （P〈 0.001）. CONCLUSION： Our results show the considerable psychological and emotional burden that a diagnosis of CHC represents, even in the absence of significant liver disease. They should be taken into account when announcing a diagnosis of CHC in order to reduce its negative effects.

Hepatitis C virus core proteins derived from different quasispecies of genotype 1b inhibit the growth of Chang liver cells

AIM: To investigate the influence of different quasispecies of hepatitis C virus (HCV) genotype 1b core protein on growth of Chang liver cells. METHODS: Three eukaryotic expression plasmids (pEGFP-N1/core) that contained different quasispecies truncated core proteins of HCV genotype 1b were constructed. These were derived from tumor (T) and non- tumor (NT) tissues of a patient infected with HCV and C191 (HCV-J6). The core protein expression plasmids were transiently transfected into Chang liver cells. At different times, the cell cycle and apoptosis was assayed by flow cytometry, and cell proliferation was assayed by methyl thiazolyl tetrazolium (MTT) assay. RESULTS: The proportion of S-phase Chang liver cells transfected with pEGFP-N1/core was significantly lower than that of cells transfected with blank plasmid at three different times after transfection (all P < 0.05). The proliferation ratio of cells transfected with pEGFP-N1/corewas significantly lower than that of cells transfected with blank plasmid. Among three different quasispecies, T, NT and C191 core expression cells, there was no significant difference in the proportion of S- and G0/G1-phase cells. The percentage of apoptotic cells was highest for T (T > NT > C191), and apoptosis was increased in cells transfected with pEGFP-N1/core as the transfection time increased (72 h > 48 h > 24 h). CONCLUSION: These results suggest that HCV genotype 1b core protein induces apoptosis, and inhibits cell- cycle progression and proliferation of Chang liver cells. Different quasispecies core proteins of HCV genotype 1b might have some differences in the pathogenesis of HCV persistent infection and hepatocellular carcinoma.

The Identification of Three Sizes of Core Proteins during the Establishment of Persistent Hepatitis C Virus Infection in vitro

Similar to Hepatitis C virus (HCV) infection in humans, HCVcc infection can also result in persistent and chronic infection. The core protein is a variable protein and exists in several sizes. Some sizes of core proteins have been reported to be related to chronic HCV infection. To study the possible role of the core protein in persistent HCV infection, a persistent HCVcc infection was established, and the expression of the core protein was analysed over the course of the infection. The results show that there are three sizes of core proteins (p24, p21 and p19) expressed during the establishment of persistent HCVcc infection. Of these, the p21 core protein is the mature form of the HCV core protein. The p24 core protein is the phosphorylated form of p21. The p19 core protein appears to be a functional by-product generated during the course of infection. These three core proteins are all localized in the cytoplasm and can be encapsidated into the HCV virion. The appearance of the p19 and p24 core proteins might be related to acute HCVcc infection and chronic infection respectively and may play an important role in the pathology of a HCV infection.

The cardiotoxicity and long-term efficacy of different doses of epirubicin in the adjuvant treatment of breast cancer patients: A case control study

Abstract Objective： The aim of the study was to observe the cardiac toxicity caused by different doses of epirubicin in the adjuvant treatment of breast cancer and to evaluate the long-term efficacy. Methods： The 180 cases of breast cancer patients received epirubicin based adjuvant chemotherapy. The patients were randomly assigned to high-dosage group （90 rag/m^2）, medium-dosage group （70 mg/m^2） and low-dosage group （50 rag/m^2）, the primary endpoint was cardiac toxicity. The secondary outcomes were the 5-year overall survival （OS） and 5-year disease-free survival （DFS）. Results： During chemo- therapy, the clinical symptoms such as palpitation, dyspnea and paroxysmal nocturnal dyspnea occurred in 6 patients with the high-dosage group, 4 patients with the medium-dosage group and 3 patients with the low-dosage group. The number of patients who had changed in electrocardiogram （ECG） was 7, 5 and 4 in three groups, respectively. The echocardiographic showed each group had only one case with LVEF 〈 50%, there was no significantly difference （P 〉 0.05）. In the three groups, the 5-year DFS rates were 73.3% （44/60） in high-dose group, 53.3% （32/60） in medium-dose group and 41.6% （25/60） in low dose group. The 5-year OS rates were 85.0% （51/60）, 68.3% （41/60） and 58.3% （35/60） in three groups, respectively. The differences were statistically significant （P 〈 0.05）. Conclusion： The high-dose epirubicin in adjuvant chemotherapy with CEF （cyclophosphamide, epirubicin and fluorouracil） regimen could improve the 5-year OS rate and 5-year DFS rate on patients of breast cancer. The cardiotoxicity was mild-moderate and well tolerated.

EXPRESSION OF PERFORIN AND FAS LIGAND IN INFILTRATING CELLS OF MURINE MYOCARDIUM INFECTED WITH COXSACKIEVIRUS B3

Purpose. To clarify the role of perforin-and Fas ligand (L)-mediated cytotoxicity pathogenesis of viral myocarditis. Materials and methods. Forty balb/c mice were randomly divided into experimental group (n = 20) and control group (n = 20), and inoculated intraperitoneally with coxsackievirus B3(CVB3) and Eagle’s solu- tion without CVB3, respectively. The mice were sacrificed and their hearts were removed at day 7 post-in- oculation. Expression of perform and FasL were detected with immunohistochemistry, reverse transcription-polymerase chain reaction (RT-PCR) and in situ hybridization. Results. (1)Perform-and FasL-positiye cells were demonstrated in experimental murine hearts by im- munohistochemistry, however, no cells were discovered in control murine hearts; (2) The examination of RT-PCR showed the positive ratios of perform and FasL mRNA in myocardium were significantly higher in experimental group (100% and 100 % ) than that in control group (20% and 30 %, P<0.05); (3)Positive signals of perform and FasL mRNA were found in myocardium of all the experimental mice by in situ hybridization, but nothing was detected in control group. Conclusion. Perform and FasL can be expressed in infiltrating cells in murine myocardium with acute myocarditis caused by CVB3, suggesting perform and FasL might play an important role in pathogenesis of viral myocarditis.

GENE TRANSFER INTO PORCINE MYOCARDIUM VIA PERICARDIAL CAVITY BY HOMEMADE EASY PUNCTURE DEVICE

Objective To explore the feasibility and safety of gene transfer into porcine myocardium via the pericardial cavity by a homemade easy device. Methods Replication-deficient recombinant adenoviral vector carrying LacZ report gene (Ad-LacZ) was constructed by the calcium phosphate precipitation method. Twelve healthy Chinese mini-swine were randomly divided into experimental group (n=6) and control group (n=6). Acute myocardial infarction (AMI) model was established by balloon occlusion of the distal part of D1 branch of left anterior descending (LAD) artery, at the same time the intra-pericardial cavity injections were performed through the small incision of the abdominal wall below the xyphoid appendix using a homemade device. Then gene transfer was performed using a central venous catheter. The pericardium was pretreated with injection of a mixture of collagenase (1 200 U) and hyaluronidase (3 000 U) in both groups. Then 2.0×109 plaque formation unit (PFU) Ad-LacZ was injected into the pericardial cavity in experimental group, while 1 mL of normal saline was injected in the control group. The β-galactosidase activity detection and X-gal staining of the ischemic myocardium were performed on the 3rd, 7th, and 28th day after injection. Results The LAD artery was occluded completely and infarction and ischemia were detected by histological assessment. In experimental group, the X-gal staining positive cells and β-galactosidase activity quantification were detectable on the 3rd day after injection, increased markedly on the 7th day, and then declined on the 28th day. The transfer efficiencies indicated by the positive myocardial cells were 16.7%, 45.6%, 22.8% on the 3rd, 7th, 28th day, respectively. In control group, no positive cells and β-galactosidase activity were observed. Conclusion Adenovirus can be transferred into ischemic myocardium and express target gene in the AMI model for four weeks with the homemade easy device via pericardial cavity pretreated by collagenase and hyaluronidase.