Host genes involved in lipid metabolism are differentially affected during the early stages of hepatitis C virus (HCV) infection.Here we demonstrate that artificial up-regulation of fatty acid biosynthesis has a positive effect on the replication of the HCV full-length replicon when cells were treated with nystatin.Conversely,the HCV RNA replication was decreased when fatty acid biosynthesis was inhibited with 25-hydroxycholesterol and PDMP(D-threo-1-phenyl-2-decanoylamino-3- morpholino-1-propanol).In agreement with these results,the expression level of GlcT-1(ceramide glucosyltransferase),a host glucosyltransferase in the first step of GSL (glycosphingolipid) biosynthesis,was found to be closely associated with the expression and replication of HCV RNA.On the other hand,the viral RNA can also activate GlcT-1 in the early stage of viral RNA transfection in vitro.To identify viral factors that are responsible for GlcT-1 activation,we constructed ten stable Vero cell lines that express individual HCV proteins.Based on the analyses of these cell lines and transient transfection assay of the GlcT-1 promoter regions,we conclude that HCV proteins,especially NS5A and NS5B,have positive effects on the expression of GlcT-1.It is possible that NS5A and NS5B stimulate transcription factor(s) to activate the expression of GlcT-1 by increasing its transcription level.

Objective：To set up a system for the culture of Dendrobium chrysotoxum in vitro. Method：Tissue culture，fire fly luminescence and phenol-H2SO4 method. Result：The embryo could germinate with or without light, the MS, 1/2MS, B5, N6 mediums are suitable to the growth and the differentiation of sprout with light, 0.5　mg*L－1NAA and 1　mg*L－16-BA, and ATP have regular changes, the content of polysaccharide was 2.833% in plant and 7.254% in sprout. Conclusion：The light has no effects on the embryo germination, but the phytohormone，nitrogen source and organized elements are important to the growth and differentiation of the sprout which should be transferred to the MS, 1/2MS, B5, N6 mediums in time supplemented with NAA与6-BA, ATP may be served as the dynamic indication of nourishment demand in the plant. The content of polysaccharide in the sprout is higher and can be utilized.

OBJECTIVETo observe the effect of Qingfei Peiyuan Micro-pill (QPM) on HIV/AIDS patients with pulmonary infection of phlegm heat obstructing lung syndrome (PHOLS).

METHODSTotally 141 HIV/AIDS patients with pulmonary infection of PHOLS were randomly assigned to the treatment group (94 cases) and the control group (47cases). On the basis of Western medicine, patients in the treatment group took QPM. The therapeutic course for all was 28 days. The improvement of symptoms and signs was observed. The body temperature (BT), chest X ray, and white blood cells (WBCs) were detected.

RESULTSThe Chinese medical syndrome score was lower in the treatment group than in the control group at the 7th, 21st, and 28th day of treatment, showing statistical difference (P < 0.05). The efficacy was better in the treatment group than in the control group at the 7th, 21st, and 28th day of treatment, showing statistical difference (P < 0.05). The BT was lower in the treatment group than in the control group on the 7th day. There was no statistical difference in the patient number with normal WBCs on the 7th day (P > 0.05). But there was statistical difference in the patient number with normal WBCs on the 14th, 21st, and 28th day of treatment (P < 0.05). There was no statistical difference in the patient number with normal chest X ray on the 7th and 28th day of treatment (P > 0.05). But there was statistical difference in the patient number with normal chest X ray on the 14th and 21 st day of treatment (P < 0.05).

CONCLUSIONQPM had certain complementary effect on HIV/AIDS patients with pulmonary infection of PHOLS.

Acquired Immunodeficiency Syndrome ; complications ; Adult ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Respiratory Tract Infections ; complications ; drug therapy ; Treatment Outcome

AIMTo detect the hepatotoxic pyrrolizidine alkaloids (HPA) in the genus Ligularia Cass..

METHODSThe alkaloid extracts of Ligularia plant materials were detected and analyzed by the method of combination of TLC, and LC/MSn.

RESULTSAmong 22 species of Ligularia Cass., HPA were detected in 18 species with LC/MSn, and no HPA was detected in the remaining 4 species.

CONCLUSIONHPA was first detected with LC/MSn in L. tongelensis and other 15 species of Ligularia Cass.; HPA from these plants should be isolated, separated and identified and it is necessary to study the activities and toxicities of the HPA. The types and kinds of HPA from different species and sources are different, they should be detected separately.

Asteraceae ; chemistry ; classification ; Chromatography, Thin Layer ; Molecular Structure ; Plants, Medicinal ; chemistry ; Pyrrolizidine Alkaloids ; analysis ; chemistry ; Species Specificity ; Spectrometry, Mass, Electrospray Ionization

OBJECTIVETo investigate the changes of gene expression in rat neurons induced by 1.8 GHz radiofrequency electromagnetic fields (RF EMF) and to screen for the RF EMF-responsive genes.

METHODSNewly-born SD rats in 24 hours were sacrificed to obtain cortex and hippocampus neurons. The cells were divided randomly into two groups: the experiment group (the irradiation group) and the control group (the false irradiation group). In the irradiation group, after twelve days' culture, neurons were exposed to 1.8 GHz RF EMF modulated by 217 Hz at a specific absorption rate (SAR) of 2 W/kg for 24 hours (5 minutes on/10 minutes off) while in the false control group, the neurons were put in the same waveguide as in the irradiation group, but were not exposed to any irradiation. The total RNA was isolated and purified immediately after exposure. The affymetrix rat neurobiology U34 assay was used for detecting the changes in gene expression profile according to the manufacturer's instruction. RF EMF-responsive candidate gene was confirmed by using ribonuclease protection assay (RPA).

RESULTSAmong 1200 candidate genes, the expression levels of 34 genes were up or down regulated. Microtubule associated protein 2 (Map2) gene was selected as the candidate and subjected to further analysis. RPA data clearly revealed that Map2 was statistically significantly up-regulated after neurons were exposed to the RF EMF (P < 0.05).

CONCLUSIONThe modulation of gene expression and function of Map2 as a neuron specific cytoskeleton protein is crucial to maintain the normal framework and function of neurons. The finding that 1.8 GHz RF EMF exposure increases the expression of Map2 might indicate some unknown effects of RF EMF on neurons.

Animals ; Animals, Newborn ; Cell Phone ; Cells, Cultured ; Dose-Response Relationship, Radiation ; Down-Regulation ; Electromagnetic Fields ; Female ; Gene Expression ; radiation effects ; Male ; Microtubule-Associated Proteins ; biosynthesis ; genetics ; Neurons ; metabolism ; radiation effects ; Radio Waves ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Up-Regulation

Recently, the interactions between hepatitis C virus (HCV) genes and the host cell factors were the focus of this field. Cell factors in the different biochemical pathway were approved to be interfered when HCV infection. To make sure which HCV gene(s) was the major factor during the interaction process, ten eukaryotic expression plasmids containing different functional genes of HCV: Core, E1, E2, p7, NS2, NS3, NS4A, NS4B, NS5A and NS5B were transfected into the CHO-K1 cells respectively. Then ten stable cell lines expressing different HCV functional proteins were constructed under the selective pressure of G418. DNA and mRNA of the HCV genes were both detected by PCR and RT-PCR respectively in the corresponding stable cell lines, freezation and anabiosis would not lose the HCV genes. Besides, the El, E2 and NS5B proteins were detected by Western-blot which demonstrated that the HCV genes have formed stable expression in the host cells. The activity of UDP-glucose ceramide glucosyltransferase (UGCG) in the stable cell lines increased in different degree by TLC assay. For example, the activity of UGCG in CHO-K1-E2 and CHO-K1-p7 was doubled according to the control cells,and in CHO-K1-NS2 and CHO-K1-NS5A was about 1.6 times compared with the control cells. The establishment of the stable cell lines containing different single HCV gene will provide foundation for investigating the interactions between the virus and the host factors, and for the filtration of antiviral medicine.
Animals ; CHO Cells ; Cricetinae ; Cricetulus ; Glucosyltransferases ; biosynthesis ; metabolism ; Hepacivirus ; genetics ; metabolism ; Transfection ; Viral Envelope Proteins ; biosynthesis ; genetics ; Viral Nonstructural Proteins ; biosynthesis ; genetics ; Viral Proteins ; biosynthesis ; genetics

OBJECTIVETo generate a gene delivery plasmid carrying the dominant negative form of the protein phosphatase 2A catalytic subunit a (DN-PP2Aca) driven by a hepatocellular carcinoma (HCC) tissue-specific promoter and investigate its ability to inhibit growth of cultured hepatoma cells.

METHODSThe gene delivery plasmid was constructed by PCR-amplifying DN-PP2Aca from wild-type PP2Aca using site-directed mutagenesis and then ligating the sequence-verified amplicon downstream of an alpha-fetoprotein enhancer and phosphoglycerate kinase promoter (AFpg) in the luciferase reporter vector pGL3-Basic. Following transfection into two AFP+ hepatoma cell lines (HepG2 and HepG3) and two AFP- hepatoma cell lines (SK-HEP-1 and L02), the transcriptional activity of the AFpg-driven DN-PP2Aca plasmid was tested using luciferase reporter gene assay and western blotting. The effect on cell growth was tested using MTT assay. Between group differences were assessed by t-test.

RESULTSThe AFpg-driven DN-PP2Aca plasmid showed high transcriptional activity and protein expression in both HepG2 and Hep3B cells. At 72 h after transfection, the proliferation capacities were repressed by 42.65%+/-3.99% (P = 0.0002) and 39.87%+/-3.91% (P = 0.0002) in AFP+ HepG2 and Hep3B cells, respectively (vs. untransfected). In contrast, the plasmid was transcriptionally inactive in and had no effect on proliferation of AFP- cells.

CONCLUSIONThe AFpg-driven DN-PP2Aca plasmid exhibits selective cytotoxicity against AFP+ hepatoma cells, and may represent a useful gene therapy strategy to treat HCC.

Carcinoma, Hepatocellular ; genetics ; metabolism ; Enhancer Elements, Genetic ; Genetic Therapy ; Genetic Vectors ; Hep G2 Cells ; Humans ; Liver Neoplasms ; genetics ; metabolism ; Mutation ; Promoter Regions, Genetic ; Protein Phosphatase 2 ; genetics ; alpha-Fetoproteins ; genetics

OBJECTIVE We have recently reported that cysteinyl leukotriene (CysLT) signaling plays an important role in microglial interleukin (IL)-1β secretion and subsequent neurotoxicity. The present study aimed to examine microglial morphological changes and the upstream molecular underlying IL-1βproduction in CysLT receptor agonist leukotriene D4 (LTD4)-treated BV2 microglia in vitro. METHODS Twenty-four hours after murine microglial BV2 cells were stimulated with LTD4 (1-100 nmol·L- 1), the cell proliferation and morphology were observed. The expression level of cysteinyl aspartate-specific protease 1 (CASP1) protein was measured by Western blotin BV2 cells. In addition, BV2 cells were pretreated with or without CysLT1 receptor antagonist montelukast for 1 h and the effects of monte-lukaston LTD4-stimulated microglial activation and CASP1 expression were evaluated. RESULTS The number of BV2 cells had an increasing tendency after 24 h treatment with LTD4, but no significant differences were observed between the control and LTD4-treated cells (P>0.05). Under basal and resting conditions, BV2 microglial cells displayed a ramified morphology. However, LTD4 at 100 nmool · L- 1 drove microglial morphological changes from a ramified towards an amoeboid shape. The expression of CASP1 protein was significantly upregulated in 100 nmool·L-1 LTD4-treated BV2 microglia (P<0.01). Furthermore, pretreatment with CysLT1 receptor antagonist montelukast prevented cell morphological changes and suppressed the increased CASP1 expression in LTD4-treated BV2 cells (P<0.05). CONCLUSION CysLT receptor agonist LTD4 induces morphological changes and CASP1 expressionin BV2 microglia, which can be inhibited by CysLT1 antagonist. These results suggest the involvement of CysLT signaling in microglial morphological changes and CASP1 expression.

Objective:To explore the efficacy of negative pressure suction curettage combined with InMode diamond carving in the treatment of axillary osmidrosis.Methods:A total of 200 patients with bromhidrosis, 89 males and 111 females, aged (27.85±7.98) years, were prospectively included in the study from February 2022 to February 2023 in Huamei Zixin Medical Cosmetology Hospital in Sichuan Province. According to different treatment methods, the patients were divided into 2 groups with 100 cases in each group by random number table method. The control group received negative pressure suction curettage, and the experimental group received combination with InMode diamond carving. After follow-up for one year, the efficacy, recurrence rate and incidence of adverse reactions were compared between the two groups.Results:In the experimental group, 94 cases were cured, 6 cases were effective, 0 cases were ineffective; the effective rate was 100% (100/100). In the control group, 90 cases were cured, 5 cases were effective and 5 cases were ineffective; the effective rate was 95% (95/100). The effective rate of experimental group was higher than that of control group, and the difference was statistically significant (χ 2=5.128, P=0.024). In the experimental group, no cases relapsed, and the recurrence rate was 0. In the control group, 5 cases relapsed, and the recurrence rate was 5.3% (5/95). The recurrence rate of experimental group was lower than that of control group, and the difference was statistically significant (χ 2=5.390, P=0.020) In control group, 5 cases of subcutaneous hematoma, 1 case of scar contracture, 7 cases of scar hyperplasia, and 1 case of skin necrosis occurred; the incidence of adverse reactions was 14.0% (14/100). In the experimental group, 2 cases of subcutaneous hematoma, 5 cases of scar contracture and 5 cases of scar hyperplasia occurred; the incidence of adverse reactions was 12.0% (12/100). There was no significant difference in the incidence of postoperative adverse reactions between the two groups (χ 2 = 0.177, P=0.674). Conclusions:Negative pressure suction curettage combined with InMode diamond carving has better curative effect and lower recurrence rate in the treatment of axillary osmidrosis.

OBJECTIVETo provide scientific basis for quality control of Lindera aggregata.

METHODHPLC analytical method was established using a Lichrospher C18 column and acetonitrile-water (56:44) as the mobile phase, detected at 235 nm.

RESULTThe linear range of linderane is between 0.0642 - 0.5774 microg, the average recovery was 98.4%, RSD1.7% (n = 9).

CONCLUSIONContents of linderane in commercially available and collected samples were from 0.028% to 0.123% and from 0.056% to 0.222% respectively.

Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; analysis ; Lindera ; chemistry ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; Sesquiterpenes ; analysis