The genetic analysis of herpesviruses has been a constant challenge, due to the large, complex genomes of herpesviruses and mutagenesis of viral genes by conventional recombination methods in cell culture. Recently, a completely new approach for full-length infectious clones of herpesviruses based on bacterial artificial chromosomes (BACs) has been developed. This technique allows the maintenance, propagation and genetic modification of the viral genome as a BAC plasmid in E.coli, thus making the procedures fast, safe and effective in prokaryotic cells. This technique also makes it possible for the reconstitution of viral progeny or mutants by transfection of the BAC plasmid into eukaryotic cells, thereby facilitating the analysis of viral gene functions in the context of genome. In this presentation, Epstein-Barr virus was used as an example to describe the principle, establishment of the technique and mutation introduction into the BAC plasmid, and to discuss the perspective in the use of BAC-cloned herpesviruses.

In order to provide scientific basics for exploitation and sufficient application of Dendrobium officinale leaves resources, the phenol-sulfuric acid method was applied to determine the polysaccharide content. The monosaccharides were derivated by PMP and the derivatives were identified by HPLC-DAD-ESI-MS(n) and the contents of mannose and glucose were determined simultaneously. Similarity evaluation system for chromatographic fingerprint of traditional Chinese medicine (2004A) was employed to generate the mean chromatogram and similarity analysis of the samples was carried out. The results demonstrated that polysaccharide content, monosaccharide compositions and composition ratio had an obvious difference between stems and leaves. The polysaccharide content of stems was higher than that of leaves. Monosaccharide composition in leaf was significantly different from that in stem. The polysaccharide from stems was composed of mannose and glucose, however the polysaccharide of leaves was acid heteropolysaccharide and was mainly composed of five monosaccharides, including mannose, galacturonic acid, glucose, galactose and arabinose. The similarity value of the 14 batches was above 0.9, indicating that similarity of fingerprints among different samples was high. The study can provide evidence for expanding the medicinal parts of D. officinale.
Chromatography, High Pressure Liquid ; Dendrobium ; chemistry ; Mass Spectrometry ; Plant Extracts ; chemistry ; Plant Leaves ; chemistry ; Plant Stems ; chemistry ; Polysaccharides ; chemistry

Epstein-Barr virus (EBV) is a human herpesvirus associated with important human diseases, including infectious mononucleosis syndrome, malignant lymphoma, and nasopharyngeal carcinoma. The mechanism of EBV entry into host cells remains a subject of intensive research. After decades of study, researchers have identified several key proteins and different patterns of EBV intrusion into host cells. The viral surface glycoproteins, gp350/220, gp42, gB, gH, and gL, are involved in interactions with the CR2 receptor on the surface of B lymphocytes during viral entry. However, the majority of epithelial cells lack CR2 receptor expression, which makes viral invasion much more complex than in B lymphocytes. Three different models have been proposed to explain how EBV enters epithelial cells: (1) "transfer of infection", mediated by B lymphocytes or Langerhans cells; (2) EBV utilizes its own proteins during the process of fusion with the cell membrane; and (3) progeny virions arising from EBV-infected epithelial cells cross lateral membranes into adjacent epithelial cells. This review will discuss the relevant mechanism of viral entry into B lymphocytes and epithelial cells during EBV infection.
Animals ; B-Lymphocytes ; virology ; Epithelial Cells ; virology ; Epstein-Barr Virus Infections ; virology ; Herpesvirus 4, Human ; genetics ; physiology ; Humans ; Viral Proteins ; genetics ; metabolism ; Virus Internalization

OBJECTIVETo study the effect of Qianyang Recipe (QYR) on the Gan-yang hyperactivity syndrome (GYHS), the blood pressure, and correlated vascular regulatory factors of hypertension rat.

METHODSThirty SD rats were randomly divided into the normal control group, the model group, and the QYR group, ten in each. Hypertension rat model of GYHS was prepared using Aconiti Praeparata Decoction plus ephedrine plus salt water. Rats in the QYR group orally took QYR physic liquor, while distilled water was given to rats in the normal control group and the model group. They were medicated for 28 successive days. The facial temperature, the grip strength, and the systolic pressure were determined once every 7 days. Rats' irritable degree and feather color were observed and recorded once every 14 days. After the last administration the plasma renin (PR), angiotensin II (Ang II), aldosterone (ALD), atrial natriuretic peptide (ANP), calcitonin gene-related peptide (cGRP) were determined.

RESULTSCompared with the model group of the same phase, the facial temperature of rats in the QYR group significantly decreased on the 14th, 21th and 28th day after administration. The systolic pressure obviously decreased on the 21st day after administration. On the 28th day after administration symptoms such as irritability, dry hair were improved, and the Ang II level decreased. There was significant difference in all these changes (P<0.05, P<0.01).

CONCLUSIONSQYR could relieve GYHS rats' symptoms such as facial hotness, irritability, dry hair, and so on, and decrease the systolic pressure. Decreased Ang II level might be one of its mechanisms.

Aldosterone ; blood ; Angiotensin II ; blood ; Animals ; Atrial Natriuretic Factor ; blood ; Calcitonin Gene-Related Peptide ; blood ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Hypertension ; blood ; diagnosis ; Male ; Medicine, Chinese Traditional ; Rats ; Rats, Sprague-Dawley ; Renin ; blood

OBJECTIVETo observe the effect of Polygonatum sibiricum on Yin deficiency model rats induced by long-term overload swimming.

METHODExcept for the normal group, all of the remaining rats performed the long-term overload swimming for eight weeks, with five days every week and once every day, to establish the Yin deficiency model. The daily swimming time increased from 10 min to 180 min at the end of the 7th week, with the water depth of 60 cm and the water temperature at 30 degrees C. After the success of the modeling, the rats were orally administered with different doses of aqueous extracts from P. sibiricum (2.5, 10 g x kg(-1)) for eight weeks. After the final administration, their blood were collected from orbits to measure immunoglobulin A, G and M (IgA, IgG, IgM), interleukin 2 and 6 (IL-2, IL-6) and cAMP, cGMP contents in plasma General behavioral indicators (weight, facial temperature, pain threshold and holding power) of rats were observed during the drug administration.

RESULTCompared with the model control group, aqueous extracts from P. sibiricum was given for eight weeks to significantly increase the rat weight and holding power of Yin deficiency model rats, decrease the facial temperature and the sensitivity of pain threshold, and increase IgA, IgG, IgM and IL-6 content and IgG content in serum, but without statistical difference. Aqueous extracts from P. sibiricum (10 g x kg(-1)) could also increase IL-2 content in serum, and decrease cAMP content and cAMP/cGMP ratio.

CONCLUSIONP. sibiricum could improve the general behavioral indicators (weight, holding power, pain threshold and facial temperature), immunologic functions (IgA, IgG, IgM) and cyclic nucleotide (cAMP, cAMP/cGMP), so as to ameliorate such Yin deficiency symptoms as dysphoria in chestpalms-soles, weight loss, soreness and weakness of waist and knees, immunologic dysfunction and cyclic nucleotide system disorders.

Animals ; Body Weight ; drug effects ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Humans ; Male ; Polygonatum ; chemistry ; Rats ; Rats, Sprague-Dawley ; Swimming ; Yin Deficiency ; drug therapy ; physiopathology

OBJECTIVETo verify the regulatory effects of acupuncture on exercise tolerance in patients with chronic obstructive pulmonary disease (COPD) at stable phase.

METHODSThirty cases of COPD were randomly divided into a treatment group (16 cases) and a placebo group (14 cases). Based on specified aerobic exercise, acupuncture was applied in the treatment group and placebo acupuncture was used in the placebo group. The acupoints included Danzhong (CV 17), Rugen (ST 18), Guanyuan (CV 4), Zhongwan (CV 12), Tianshu (ST 25) and so on. The needle did not penetrate into the skin for the placebo group. The treatment was required for 2 to 3 times per week for totally 5 weeks. The indices of exercise tolerance, including 6-min walking distance (6-MWD), exercise time, maximum oxygen uptake (VO2max) forced expiratory volume in 1 second (FEV1)/forced vital capacity (FVC), maximum ventilatory volume (MVV), St. George respiratory questionnaire (SGRQ) were observed in two groups before and after treatment.

RESULTS(1) Exercise tolerance: the differences of 6-MWD and exercise time were statistically significant between groups, which were more superior in the treatment group (both P<0.01); the VO2max was significantly increased after treatment in the treatment group (P<0.05), but there was no difference between two groups (P>0.05). (2) Pulmonary ventilation function: the differences of FEV1%, FEV1/FVC and MVV% were statistically significant between groups, which were more superior in the treatment group (P<0.05, P<0.01); (3) SGRQ: the SGRQ was significantly improved after treatment in the treatment group (P<0.05), but there was no difference between two groups (P>0.05).

CONCLUSIONThe acupuncture could improve the exercise tolerance in patients with chronic obstructive pulmonary disease at stable phase, and shorten the onset time of aerobic exercise. Besides, acupuncture combined with aerobic exercise could effectively improve the pulmonary function.

Acupuncture Points ; Acupuncture Therapy ; Aged ; Exercise Tolerance ; Female ; Humans ; Male ; Middle Aged ; Pulmonary Disease, Chronic Obstructive ; physiopathology ; therapy

OBJECTIVETo study the endothelioid differentiation effect of Epimedin C on murine embryonic mesenchymal stem cells (C3H/10T1/2).

METHODSC3H/10T1/2 cells were cultivated in vitro. The cytotoxicity of Epimedin C at different concentrations was determined by MTT assay and crystal violet assay. Morphological changes were observed under microscope after treated with Epimendin C. The effect of Epimendin C on the cell cycle distribution was determined by flow cytometry. mRNA expression levels of endothelial markers, such as CD31, CD34, vascular endothelial zinc finger 1 (Vezf1), angiopoietin 1 (Ang1), and angiopoietin 2 (Ang2) were detected by semi-quantitative PCR. Protein expression levels of platelet endothelial adhesive molecule 1 (CD31), ecto-5'-nucleotidase (CD73), endothelial cell specific molecule-1 (ESM-1), and integrin β5 were determined by immunocytochemical (IHC) staining.

RESULTSEpimedin C could not affect the survival rate of C3H/10T1/2 cells at 1-30 μmol/L. Its cell cycle distribution was not significantly changed after treated by 30 μmol/L Epimedin C for 24 h. C3H/10T1/2 cells were differentiated to vascular endothelial cells by Epimedin C treatment, with significant morphological changes (whirlpool-like structure). PCR results indicated that mRNA levels of classic endothelial mark- ers, namely CD34, Vezf1, Ang1, and Ang2 were significantly increased in C3H/10T1/2 cells after treated with Epimedin C for 5 days (P < 0.05, P < 0.01). Protein expression levels of CD31, CD73, and ESM-1 were also positively expressed after treated with Epimedin C for 5 days, showing statistical difference when compared with those of the control group (P < 0.01, P < 0.05).

CONCLUSIONEpimendin C could induce C3H/10T1/2 cells to differentiate into endothelioid cells.

Animals ; Cell Differentiation ; drug effects ; Cell Line ; Cells, Cultured ; Flavonoids ; pharmacology ; therapeutic use ; In Vitro Techniques ; Mesenchymal Stromal Cells ; physiology ; Mice ; RNA, Messenger

In order to investigate the anti-proliferative effects of triptolide (TP) on 4T1 mice breast cancer cell line in vitro and in mouse model, as well as the possible mechanisms, we detected the effect of TP on cell proliferation by MTT assay or Crystal Violet Staining in our research. Flowcytometry combined with FITC-Annexin V/PI staining were used for detecting TP induced 4T1 cell apoptosis. The protein expression of ERalpha, p-ERalpha, ERbeta, p-ERbeta, ERK, p-ERK, p38, p-p38, SAPK/JNK, and p-SAPK/JNK was tested by western blotting. We also compare TP with chemotherapy drug doxorubicin in 4T1 tumor bearing BLAB/c mice model, the Xenogen bioluminescence imaging, H&E, and IHC result indicated that TP exhibits an anticancer proliferation activity. As a result, TP in 100, 10, 1, 0.1 micromol x L(-1), all inhibited the proliferation of 4T1 cells by MTT assay and Crystal Violet Staining. TP which concentrations is 10, 1, 0.1 micromol x L(-1) could induce the apoptosis of 4T1 cells and reduce the cell proliferation. TP in 200 microg x kg(-1) could inhibit the tumor growth in vivo. The anticancer proliferation of TP was involved in its effect on reducing expression of ERalpha, p-ERalpha, ERbeta, and p-ERbeta, but nothing to do with the activation of MAPK signaling pathway.
Animals ; Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; Diterpenes ; pharmacology ; therapeutic use ; Down-Regulation ; drug effects ; Epoxy Compounds ; pharmacology ; therapeutic use ; Female ; Lung Neoplasms ; secondary ; Mammary Neoplasms, Experimental ; drug therapy ; metabolism ; pathology ; Mice ; Mice, Inbred BALB C ; Phenanthrenes ; pharmacology ; therapeutic use ; Phosphorylation ; drug effects ; Receptors, Estrogen ; metabolism ; Tumor Burden ; drug effects

Objective:To explore clinical and histopathological characteristics of primary biliary cirrho-sis-autoimmune hepatitis overlap syndrome.Methods:Clinical data and pathological findings of 10 pa-tients were reviewed.Results:Serum glutamine transpeptidase,alkaline phosphatase levels,alaninetransaminase,aspartate transaminase,serum IgG and IgM were elevated in all the patients.They were allpositive for anti-mitochondrial antibody and AMA-M2.Nine patients were positive for anti-nuclear anti-body and one patient was positive for anti liver-kidney microsome antibody.Liver biopsies in these pa-tients revealed:ten patients had bile duct lesion,hepatitis activities ranged from moderate to severe,andfibrosis ranged from S1 to S3.Conclusion:PBC-AIH overlap syndrome is mostly found in middle-agedwomen.It has the clinical and histopathological characteristics of both PBC and AIH.Accurate andprompt diagnosis of overlap syndrome patients should be based on the clinical presentation,biochemicaland immune indexes,and hepalic pathological changes.

Based on the Bacillus sp., isolated from Lop Nur Desert, the technology of separation and purification and the antioxidant effect were studied. After centrifugation and vacuum filtration, the deproteinization of supernatant was operated with Sevag reagent. The crude exopolysaccharide (EPS) was obtained by precipitation with ethanol. The optimum conditions for the isolation were as follow: pH 7.0, temperature 4?C, time 1.5 h, and material to ethanol ratio 1: 4. Dissolved in water, the crude EPS was fractional separated on activated carbon column (1.5 cm ? 24 cm), eluted with distilled water, 60% ethanol, 95% ethanol, and the main fraction was collected. Then the EPS was purified on Sephadex G-100 gel column, eluted with NaCl (0.2 mol/L). Fractions (4 mL, each) were also combined according to total sugar by phenol-sulfuric acid method and protein content was determined by Coomassie brilliant blue. The results showed that EPS was relatively homogeneous glycoprotein. The data of antioxidation in vitro showed that the EPS had a high antioxidant activity, which could quench hydroxyl radical, superoxide radical and had antilipid peroxidation activity. All of these indicated that EPS was a good natural antioxidant.