RASSF2A Promoter Methylation in Hepatitis B Virus-related Hepatocellular Carcinogenesis and Its Correlation with Elevated Serum α-Fetoprotein Level

Loss of the RASSF2A expression induced by methylation-mediated silencing has been reported in several human cancers, but the methylation status of RASSF2A in hepatocellular carcinoma （HCC） is rarely studied so far. In this study, we investigated the RASSF2A expression and its methylation status in a cohort of 45 hepatitis B virus-associated HCC tissues and their adjacent non-carcinoma tissues by using RT-PCR and MS-PCR. Promoter methylation of RASSF2A was found in 31 （68.9%） out of 45 HCC tissues and 12 （40%） out of 30 adjacent normal tissues, respectively （P〈0.05）. The methylation status of PASSF2A was closely associated with the loss of RASSF2A expression and elevated serum α-fetoprotein level, but not significantly with clinical stage, hepatic fibrosis and K-ras mutation. It was concluded that aberrant methylation of the RASSF2A gene with the subsequent loss of RASSF2A expression plays an important role in the pathogenesis of HCC.     

Relationship between the Expression of RASSF1A Protein and Promoter Hypermethylation of RASSF1A Gene in Bladder Tumor

To investigate the relationship between the expression of RASSF1A protein and promoter hypermethylation of RASSF1A gene, RASSF1A protein expression was measured by Western blotting in 10 specimens of normal bladder tissues and 23 specimens of bladder transitional cell carcinoma （BTCC）. The promoter methylation in BTCC and normal bladder tissues was detected by methylation-specific PCR （MSP）. The results showed that the expression level of RASSF1A protein was significantly lower in BTCC tissues than that in normal bladder tissues. However, it was not correlated with its clinical stages and pathological grades. The frequency of promoter methylation of RASSF1A gene was higher in BTCC tissues than that in normal bladder tissues. In 14 patients with the aberrant promoter methylation, 13 showed loss or low expression of RASSF 1A protein. It is concluded that RASSF1A gene promoter methylation may contribute to the low level or loss of RASSF1A protein expression, the inactivation of RASSF1A gene and the genesis of BTCC. But, it may bear no correlation with its clinical stages and pathological grades.     

Transgenic mice with overexpression of human scavenger receptor A on endothelial cells

Objectives To establish a new transgenic mouse model for determining the function and role of human scavenger receptor A (SR- A) in atherosclerosis in vivo. Methods Human scavenger receptor minigene- driven mouse tie- 1 promoter was constructed and confirmed by endonuclease digestion and sequence analysis. Transgenic mice were generated via the microinjection method. PCR and Southern blot were used t o screen the positive transgenic mice. RT- PCR and immunohistochemical analysis were used to detect the level and location of human SR- AⅠ expression in transgenic mice. The activity of human SR- AⅠ was determined by morphologi c observation of aortic endothelial cells of transgenic mice under transmission electron microscopy. Results The electrophoresis assay showed the expected 4 fragments of 0. 9 kb, 1. 1 kb, 1. 2 kb and 4. 2 kb in the SmaⅠ digest and 2 fragments of 0. 8 kb and 6 . 7 kb in BglⅡ digest of plasmids pTie- 1/hSR- A. The fragment sequence of tie - 1 p romoter and human SR- A cDNA in plasmids pTie- 1/hSR- A was correct and no A TG b efore the translation initiation sites of human SR- A was found by sequence ana lysis. 561 injected and surviving embryos with the purified human SR- A minigen e were implanted into the oviducts of 19 ICR pseudopregnant mice. Among the 54 surviving pups from 13 foster mothers, 7 were identified by PCR and Sou thern blot analysis. The results of RT- PCR and immunohistochemical analysis sh owed human SR- A was specifically expressed on vascular endothelial cells of the aorta and renal artery, as well as hepatic sinusoidal endothelial cells in tran sgenic mice. Transmmion electron microscope (TEM) of aorta of transgenic mice s howed that a large number of vesicles, multivesicle bodies and swollen mitochond ria filled the plasma of endothelial cells. Conclusions A transgenic mouse model with overexpression of human SR- A in endothelial cells was successfully established. The transgene was integrated and transmitted int o the chromosome of transgenic mice. Tie- 1 promoter controlled the transgene t o express in endothelial cells in mice. Pinocytic activity of aortic endothelia l cells in transgenic mice was higher than that of C57BL/6J mice. Our studies w ill provide a new transgenic model for investigation of atherosclerosis and func tions of human SR- A.     

RASSF1A expression inhibits cell growth and enhances cell chemosensitivity to mitomycin in BEL-7402 hepatocellular carcinoma cells

Background The antitumor role of Ras association domain family 1A （RASSF1A） gene and its potential molecular mechanisms are not well understood. The objective of this study was to observe the antitumor ability of RASSF1A in hepatocellular carcinoma, and study the mechanisms of cell apoptosis induced by RASSF1A. Methods After stably transfecting a RASSFIA （wild-type or mutant） expression vector into the BEL-7402 hepatocellular carcinoma cell line, RT-PCR and Western blotting was used to detect the RASSF1A expression levels in recombinant cells. The effects of wild-type RASSF1A on cell growth were observed in vitro by analyzing cell proliferation rate, cell colony formation, and in vivo by analyzing tumorigenesis in nude mice. In addition, the effect of RASSF1A gene expression on the chemosensitivity of human hepatocellular carcinoma cells to antitumor drugs was examined by inhibition of cell proliferation and the percentage of apoptotic cells. Results Wild-type RASSF1A, not the mutant, suppressed cell growth in vitro and in vivo. Re-expression of wild-type RASSF1A could enhance the inhibition of cell proliferation and the percentage of apoptotic cells following cell treatment with mitomycin, but had no significant effect when combined with adriamycin, etoposide, 5-fluorouracil and cisplatin treatment. Conclusion Wild-type RASSF1A inhibits cell growth and enhances cell chemosensitivity to mitomycin in hepatocellular carcinoma, suggesting that RASSF1A may serve as a new target for gene therapy in hepatocellular carcinoma patients.     

The infectivity of gametocytes of Plasmodium falciparum from patients treated with artemisinin.

Twenty-seven patients with gametocytes of Plasmodium falciparum(PF) were divided into groups A,B,and C.A daily dose of 1200 mg artemisinin was given for 5 days to group A,a state dose of 750 mg of mefloquine to group B and a single dose of 750mg     

Molecular epidemiology of respiratory syncytial virus

Objective To determine the epidemiologic pattem of subgroups A and B and genotypes of respiratory syncytial virus (RSV) during two noncontinuous epidemics during 1990- 1991 and 1997- 1998 in Beijing. Methods Nasopharyngeal secretion (NPS) samples of RSV positive or RSV isolates tested by indirect immunofluorescence (IIF) assay were classified into subgroups A and B. Isolates of RSV were divided into at least six different lineages, designated NP1-NP6, by restriction mapping of the N gene. Np1, 3 and 6 were given by subgroup B isolates, while NP2, 4 and 5 were given by subgroup A isolates. Strains of subgroup A were further subdivided into six lineages SHL1-SHL6 on the basis of the SH gene sequence. SH lineages were closely related to each other and to NP1-NP6. Strains of SHL1, 3 and 4 were closely related and belonged to NP2, SHL2 and 6 to NP4, and SHL5 to NP5. Results Of 145 RSV NPS samples from the 1997- 1998 epidemic, 83 (57.2％) were of subgroup B RSV positive, 62 (42.8％) of subgroup A RSV positive. The rate of occurrence of subgroup A to B strains was about 1:1.3. Two of 10 isolates during the epidemic were subgroup A strains, whereas 8 were subgroup B strains. The rate of occurrence of subgroup A to B strains was 1:4. Eight subgroup A strains of 10 isolates from the 1990 - 1991 epidemic were dominant; the proportion of subgroup A to B strains was 4:1. With 10 RSV isolates in 1997- 1998, all 2 subgroup A strains gave N gene fragment restriction pattern NP4, and fell into SH lineage SHL2, whereas 8 subgroup B strains all belonged to NP3. All 8 subgroup A isolates from the 1990- 1991 epidemic gave pattem NP4, and fell into SHL2, while 2 subgroup B strains all belonged to NP3. The classification of subgroups A and B deduced from NP pattems corresponded to the definition of these subgroups by monoclonal antibodies. Conclusions These observations confirm that subgroups A and B or multiple lineages of RSV co circulated in Beijing, but different genome types predominated each year. Moreover, very similar viruses were isolated up to more than 5 years ago, indicating that despite apparent diversity of the subgroup A strains, the separate lineages might be relatively stable.     

Transfer of Paralytic Shellfish Toxins via Marine Food Chains：A Simulated Experiment

Objective To study the transfer of paralytic shellfish toxins (PST) using four simulated marine food chains: dinoflagellate Alexandrium tamarense→Artemia Artemia salina→Mysid shrimp Neomysis awatschensis; A. tamarense→N. awatschensis; A. tamarense→A. salina→Perch Lateolabrax japonicus; and A. tamarense→L. japonicus. Methods The ingestion of A. tamarense, a producer of PST, by L. japonicus, N. awatschensis, and A. salina was first confirmed by microscopic observation of A. tamarense cells in the intestine samples of the three different organisms, and by the analysis of Chl.a levels in the samples. Toxin accumulation in L. japonicus and N. awatschensis directly from the feeding on A. tamarense or indirectly through the vector of A. salina was then studied. The toxicity of samples was measured using the AOAC mouse bioassay method, and the toxin content and profile of A. tamarense were analyzed by the HPLC method. Results Both A. salina and N. awatschensis could ingest A. tamarense cells. However, the ingestion capability of A. salina exceeded that of N. awatschensis. After the exposure to the culture of A. tamarense (2 000 cells·mL-1) for 70 minutes, the content of Chl.a in A. salina and N. awatschensis reached 0.87 and 0.024 μg·mg-1, respectively. Besides, A. tamarense cells existed in the intestines of L. japonicus, N. awatschensis and A. salina by microscopic observation. Therefore, the three organisms could ingest A. tamarense cells directly. A. salina could accumulate high content of PST, and the toxicity of A. salina in samples collected on days 1, 4, and 5 of the experiment was 2.18, 2.6, and 2.1 MU·g-1, respectively. All extracts from the samples could lead to death of tested mice within 7 minutes, and the toxin content in artemia sample collected on the 1st day was estimated to be 1.65×10-5 μg STX equal/individual. Toxin accumulation in L. japonicus and N. awatschensis directly from the feeding on A. tamarense or indirectly from the vector of A. salina was also studied. The mice injected with extracts from L. japonicus and N. awatschensis samples that accumulated PST either directly or indirectly showed PST intoxication symptoms, indicating that low levels of PST existed in these samples. Conclusion Paralytic shellfish toxins can be transferred to L. japonicus, N. awatschensis, and A. salina from A. tamarense directly or indirectly via the food chains.     

OBSERVATION OF THE TONGUE

Here"tongue"refers to the pulsc and muscle tissues of the tongue,thatis,the tongue proper.Through observation of the tongue the doctor canjudge the patient's state of internal organs and the trend of his condition. I.Observation of the Colour of Tongue. A normal tongue looks reddish,it is sleek,neither sticky and eely nordry.A tongue looking colourless and bloodless is called a pale tongue,whichis caused by diseases of cold factors belonging to deficiency of vital energy.A tougne looking scarlet is called red tongue,whrch is a sign of heatdiseases of typos of sthenia or deficiency of vital energy.A tongue looking     

Effects of heme precursors on CYP1A2 and POR expression in the baculovirus/Spodoptera frugiperda system

Objective：CYP1A2 and NADPH-CYP450 oxidoreductase（POR）were expressed in the baculovirus/ Spodoptera frugiperda（sf9）system.The aim of this study was to investigate the effects of heme precursors on the expression of CYP1A2 and POR.Methods：The heme precursors[δ-Aminolaevulinic Acid（5-ALA）,Fe^3＋and hemin]were introduced into the system to evaluate their effects on the expression of CYP1A2,POR and their co-expression.All the proteins were identified using immunoblotting,CO-difference spectroscopy,or cytochrome c assay.Results：In the present study,functional CYP1A2 and POR were successfully expressed in the baculovirus/sf9 system,and both of them showed high activities.Co-addition of 5-ALA and Fe^3＋significantly improved expression of CYP1A2 by about 50%compared with the addition of 5-ALA,Fe^3＋or hemin alone.Either co-addition of 5-ALA and Fe^3＋or addition of 5-ALA or Fe^3＋alone improved the POR expression level 2 fold and its activity 7-10 fold compared with control（no addition）.However,unlike CYP1A2,there was no difference between the co-addition and addition of these heme precursors alone.Different ratios of BvCYP1A2 to BvPOR also affected the co-expression of CYP1A2 and POR,with a 3：1 ratio of BvCYP1A2/BvPOR significantly increasing their co-expression.Surprisingly,the addition of 0.1 mM 5-ALA or Fe^3＋alone,but not their co- addition,could significantly improve the CYP1A2 and POR co-expression（P〈0.05）.Conclusion：5-ALA and Fe^3＋increased the expression of CYP1A2 and POR in a baculovirus/sf9 system,but the pattern of their expression was different between their expression alone and co-expression.     

Hypercalcemic crisis due to a mediastinal parathyroid cyst diagnosed by ultrasound-guided fine needle aspiration

Hypercalcemic crisis, generally accepted as serum calcium concentration greater than 3.5 mmol/L,constitues a life-threatening endocrinologic emergency,and is most frequently caused by either primary hyperparathyroidism （PHPT） or malignant diseases.Parathyroid cysts are uncommon lesions, most of that are located in the low part of the neck. By routine neck ultrasound scan investigation in a large series of 6621 patients, only 5 parathyroid cysts were detected, yielding a prevalence of 0.075% in setting of unselected patients.The parathyroid cysts in the mediastinum are much less frequently encountered, with only 106 cases reported in English literature.2,3 Moreover, less than half of these cases presented as functional with elevated serum calcium and parathyroid hormone, and only 10 cases were associated with hypercalcemic crisis.3 Herein, we present a rare case of mediastinal parathyroid cyst associated with recurrent hypercalcemic crisis, which diagnosed by ultrasound-guided fine needle aspiration （FNA）.     

细针穿刺术诊断头颈部淋巴瘤的价值

1文献来源Kuvezdic＇KG,Aurer I,Ries S,et al.FNAbased diagnosis of head and neck nodal lymphoma[J].Coll Antropol,2010,34（1）：7-12.2证据水平2b。     

细针穿刺对淋巴瘤诊断技术的价值

1文献来源Hehn ST,Grogan TM,Miller TR.Utility offine needle aspiration as a diagnostic technique inlymphoma[J].J Clin Oncol,2004,22（15）：3046-3052.     

细针穿刺细胞学检查在淋巴瘤病理诊断中的意义

＂鉴于这些肿瘤（恶性淋巴瘤）病因的未知性及多样性,本分类依据所有可获得的信息来定义疾病,包括形态学、免疫表型、遗传学及临床特征,     

细针穿刺活检在WHO分类基础上的非霍奇金淋巴瘤诊断价值

1文献来源Mourad WA,Tulbah A,Shoukri M,et al.Primary diagnosis and REAL/WHO classification of non-Hodgkin＇s lymphoma by fine-needle aspiration：     

Relevance of muscle biopsy for diagnosing multifocal motor neuropathy

Multifocal motor neuropathy （MMN） is a rare,.focal,inflammatory,demyelinating disease of the peripheral nerves with pure motor involvementJ MMN is clinically characterized by slowly progressive,asymmetric,distal,upper limb predominant weakness,in the absence of sensory disturbances） Weakness is usually multifocal and connected to a distinct motor nerve,such as the musculocutaneous nerve resulting in biceps weakness,the posterior interosseus nerve resulting in finger drop,the median,ulnar,or radial nerve resulting in dexterity problems or grip weakness,or the peroneal nerve resulting in a foot drop.Onset of clinical manifestations is between 20 and 50 years of age.The prevalence of MMN is reported as 1-2 per 100 000.2 MMN is three times more frequent in men as compared to women.     

Association between intervertebral disc degeneration and disturbances of blood supply to the vertebrae

Low back pain is a common public health problem in western industrialized societies and the world as well.Studies indicate that the prevalence rate ranges to 35%, with around 10% of patients from 12% becoming chronically disabled. It also places an enormous economic burden on society. Although the exact cause of low back pain has yet to be defined, intervertebral disc degeneration is considered a major source of it. Since patients with degenerative discs are often asymptomatic, the mechanisms of it are still unclear.     

Saffron （Crocus sativus L.） and major depressive disorder： a meta-analysis of randomized clinical trials

Due to safety concerns and side effects of many antidepressant medications, herbal psychopharmacology research has increased, and herbal remedies are becoming increasingly popular as alternatives to prescribed medications for the treatment of major depressive disorder （MDD）. Of these, accumulating trials reveal positive effects of the spice saffron （Crocus sativus L.） for the treatment of depression. A comprehensive and statistical review of the clinical trials examining the effects of saffron for treatment of MDD is warranted. OBJECTIVE： The purpose of this study was to conduct a meta-analysis of published randomized controlled trials examining the effects of saffron supplementation on symptoms of depression among participants with MDD. SEARCH STRATEGY： We conducted electronic and non-electronic searches to identify all relevant randomized, double-blind controlled trials. Reference lists of all retrieved articles were searched for relevant studies. INCLUSION CRITERIA： The criteria for study selection included the following： （1） adults （aged 18 and older） with symptoms of depression, （2） randomized controlled trial, （3） effects of saffron supplementation on depressive symptoms examined, and （4） study had either a placebo control or antidepressant comparison group. DATA EXTRACTION AND ANALYSIS： Using random effects modeling procedures, we calculated weighted mean effect sizes separately for the saffron supplementation vs placebo control groups, and for the saffron supplementation vs antidepressant groups. The methodological quality of all studies was assessed using the Jadad score. The computer software Comprehensive Meta- analysis 2 was used to analyze the data. RESULTS： Based on our pre-specified criteria, five randomized controlled trials （n = 2 placebo controlled trials, n = 3 antidepressant controlled trials） were included in our review. A large effect size was found for saffron supplementation vs placebo control in treating depressive symptoms （M ES = 1.62, P 〈 0.001）, revealing that saffron supplementation significantly reduced depression symptoms compared to the placebo control. A null effect size was evidenced between saffron supplementation and the antidepressant groups （M ES = -0.15） indicating that both treatments were similarly effective in reducing depression symptoms. The mean Jadad score was 5 indicating high quality of trials. CONCLUSION： Findings from clinical trials conducted to date indicate that saffron supplementation can improve symptoms of depression in adults with MDD. Larger clinical trials, conducted by research teams outside of Iran, with long-term follow-ups are needed before firm conclusions can be made regarding saffron＇s efficacy and safety for treating depressive symptoms.     

痘苗病毒天坛株的减毒与其作为疫苗载体研究进展

1痘苗病毒天坛株的历史天花是人类历史上最可怕的传染病之一。天花由天花病毒（variola virus）引起,人是该病毒的惟一宿主。人感染天花病毒后的死亡率达到30%~40%[1]。世界上公认的对天花最早的准确记录来自中国,晋代药学家葛洪于公元430年左右,在《肘后备急方》中对天花有清楚的描述[1]。