Objective: To clone the sequence and characterize gene expression of an expansin gene from Tetrastigma hemsleyanum, and to predict its probable function. Methods: According to the conservative region sequences of expansin gene in GenBank database, one pair of degenerate primers were designed for RT-PCR. The conservative region fragment was first amplified by RT-PCR from cDNA template of T. hemsleyanum. The full-length sequence of expansin gene cDNA (named as Th-exp) was extended by RACE (rapid-amplification of cDNA ends). Th-exp expression levels in different organs were conducted by semi-quantitative RT-PCR and fluorescence quantitative PCR. Results: The full-length cDNA sequences of Th-exp gene were obtained in 782 bp. Th-exp gene containsed a 630 bp open reading frame, which encoded a protein of 209 amino acids (GenBank accession number KP693606). The protein encoded by Th-exp habours a typical expansin structure, including eight cysteine domains in N terminal region, four conserved tryptophan domain in C terminal region, and histidine function domain in intermediate region. Blast alignment showed that Th-exp was similar to expansin genes of Vitis vinifera and Tarenaya hassleriana. Semi-quantitative RT-PCR and fluorescence quantitative PCR indicated that Th-exp could express in the leaves, stems, ordinary fine roots, and calabash-shaped roots, while the expression level in calabash-shaped roots was higher than those in other organs. Conclusion: The expansin gene (Th-exp) is cloned from T. hemsleyanum, and it could be involved in the development of root tubers in T. hemsleyanum.

Objective: To study the anti-inflammatory of phenolic acids from Lonicerae Japonicae Flos. Methods: The compounds in the EtOAc fraction were isolated and purified by silica gel and Sephadex LH-20 column chromatographies, and preparative HPLC as well. The structures were identified by various spectroscopic data such as ESI-MS, ¹H-NMR, and ¹³C-NMR data. The anti-inflammatory effects were evaluated by a in vitro model of LPS-stimulated RAW264.7 macrophages. Results: Eight phenolic acids, three iridoids, and one flavonoid were isolated from the extracts of Lonicerae Japonicae Flos and identified as 5-O- caffeoylquinic acid (1), 4-O-caffeoylquinic acid (2), 3,5-di-O-caffeoylquinic acid (3), 3,4-di-O-caffeoylquinic acid (4), 4,5-di-O- caffeoylquinic acid (5), chlorogenic acid (6), caffeic acid (7), methyl caffeate acid (8), secoxyloganin (9), secologanoside (10), sweroside (11), and luteolin (12). Compounds 1-8 showed the anti-inflammatory activity in different degrees. Conclusion: Phenolic acids are the main anti-inflammatory constituents in Lonicerae Japonicae Flos. Caffeic acid shows the strongest activity in vitro.

Lonicerae Flos known as "Jin Yin Hua" in Chinese, is commonly used in Chinese materia medica. Numerous compounds such as flavonoids, iridoids, triterpenoids, organic acids, and volatile components have been reported from this plant. This paper reviews the researches on the chemical constituents of Lonicerae Flos in recent 20 years, and provides the references for further studies.

Objective To discover radioresistance associated molecular biomarkers and its mechanism in nasopharyngeal carcinoma by protein-protein interaction network analysis.Methods Whole genome expression microarray was applied to screen out differentially expressed genes in two cell lines CNE- 2R and CNE-2 with different radiosensitivity.Four differentially expressed genes were randomly selected for further verification by the semi-quantitative RT-PCR analysis with self-designed primers. The common differentially expressed genes from two experiments were analyzed with the SNOW online database in order to find out the central node related to the biomarkers of nasopharyngeal carcinoma radioresistance. The expression of STAT1 in CNE-2R and CNE-2 cells was measured by Western blot.Results Compared with CNE-2 cells,374 genes in CNE-2R cells were differentially expressed while 197 genes showed significant differences.Four randomly selected differentially expressed genes were verified by RT-PCR and had same change trend in consistent with the results of chip assay. Analysis with the SNOW database demonstrated that those 197 genes could form a complicated interaction network where STAT1 and JUN might be two key nodes.Indeed,the STAT1-α expression in CNE-2R was higher than that in CNE-2 (t =4.96,P ＜ 0.05).Conclusions The key nodes of STAT1 and JUN may be the molecular biomarkers leading to radioresistance in nasopharyngeal carcinoma,and STAT1-α might have close relationship with radioresistance.

Objective： This study was designed to investigate the relationship between expression level of multidrug resistance-associated protein (MRP) or P-glycoprotein (P-gp) genes and chemotherapy efficacy or clinical drug resistance in the patients with malignant lymphomas. Methods： Using the methods of semi quantitative reverse transcription polymerase chain reaction (RT-PCR) and flow cytometry (FCM), The authors examined the expression level of MRP and P-gp of 46 lymphoma patients. Results： The expression level and positive rate of P-gp in the recurrent patients was higher than that in untreated patients (P<0.001). There was no difference in MRP gene expression level and positive rate between recurrent and untreated patients (P>0.05). Chemotherapeutic effective rate in P-gp positive patients (26.67％ ) was lower than that in P-gp negative patients (83.87％ )(P<0.001). While there was no difference between MRP gene positive and negative patients, their chemotherapy effective rate had no difference. Relevant analysis showed that there was no correlation between MRP and P-gp (r-0.0808, P>0.05). Conclusion： P-gp expression correlates to multidrug resistance, and is the major mechanism of clinical drug resistance of lymphomas, whereas, MRP gene does not appears to play any role in that course.

AlM: To investigate the association between the degree of anterioruveitis and related factors including inflammatory markers as well as sacroiliac joint imaging in patients with ankylosing spondylitis ( AS) .
METHODS: Anterior changes evaluated by slit lamp, erythrocyte sedimentation rate ( ESR ) , C - reactive protein ( CRP ) and magnetic resonance imaging of 55 cases with AS associateduveitis were retrospectively analyzed. A modified endotoxin-induced uveitis ( ElU ) clinical standard was used for uveitis grading. SPARCC sacroiliac scoring was used to evaluate bone edema of sacroiliac joint. The correlation between the degree of uveitis and sacroiliitis was assessed.
RESULTS: ln the 55 patients with AS, ElU grading scored 2-10, and SPARCC index scored 0-22. Further analysis showed that the severity of uveitis was significantly correlated with ESR (r=0. 869, P<0. 001) and CRP (r=0. 485, P<0. 001). The degree of anterior uveitis in AS patients was not correlated with inflammation of sacroiliac joint (r=0. 237, P=0. 081).
CONCLUSlON: Local autoimmunity of uveitis and sacroiliac joint inflammation with subsequent bone formation in AS might be mutually independent processes.

Objective To observe the effect of hypoxia reoxygenation on activity of superoxide dismutase(SOD)and MDA content as well as[Ca~(2+)],concentration and mitochondria membrane poten- tial of intestinal epithelial cell-6(IEC-6)in IEC culture medium and explore the protective effect and mechanism of serum containing Ziqi-liquid(a preparation of Chinese herbal medicine)on hypoxia reoxy- genation damaged IEC-6.Methods Hypoxia reoxygenation damage model of IEC-6 was made.SOD activity and MPA content in IEC-6 culture medium were determined by ultraviolet spectrometry after hy- poxia reoxygenation and treatment with Ziqi-liquid.Meanwhile,MMP changes and[Ca~(2+)]concentration were detected by laser scanning confocal microscopy.Results After hypoxia reoxygenation,SOD and MMP were significantly decreased,but MDA content and[ Ca~(2+)] concentration significantly increased (P＜0.01),and significantly facilitated by serum containing Ziqi-liquid.Conclusion Hypoxia reoxy- genation can damage IEC-6,but the serum containing Ziqi-liquid has significant protective effect on it.

OBJECTIVETo investigate the chemical constituents of dried whole plants of Artemisia annua.

METHODThe chemical constituents were isolated by repeated silica gel chromatography, medium pressure column chromatography, and semi-preparative HPLC, and their structures were elucidated by spectroscopic analyses and comparison of NMR data with those reported in literature.

RESULT15 compounds were isolated and identified to be 5-O-[(E)-Caffeoyl] quinic acid(l), 1,3-di-O-caffeoylquinic acid(2), 4 5-di-O-caffeoylquinic acid(3), 3, 5-di-O-caffeoylquinic acid (4), 3, 4-di-O-caffeoylquinic acid (5), methyl-3,4-di-O-caffeoylquinic acid(6), methyl-3,5-di-O-caffeoylquinic acid(7), 3,6'-O-diferuloylsucrose(8), 5'-β-D-glucopyranosyloxyjasmonic acid(9), Scopoletin(10), scoparone (11), 4-O-β-D-glucopyranosyl-2-hydroxyl-6-methoxyacetophenone (12), chrysosplenol D (13), casticin (14), chrysosplenetin(15).

CONCLUSIONCompounds 2, 6, 8 and 9 are obtained from the Artemisia genus for the first time. Compounds 7 and 15 are obtained from this plant for the first time.

Artemisia annua ; chemistry ; Chromatography, Gel ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Flavonoids ; chemistry ; isolation & purification ; Medicine, Chinese Traditional ; Plants, Medicinal ; Quinic Acid ; analogs & derivatives ; chemistry ; isolation & purification ; Silica Gel

OBJECTIVETo investigate the outcome of in vitro fertilization and embryo transfer (IVF-ET) in couples with the husband positive for chronic infection of hepatitis B virus (HBV).

METHODSThis study involved 102 infertile couples receiving IVF-ET with the husbands(but not the wives) positive for hepatitis B surface antigen (HBsAg), and another 204 couples negative for HBsAg receiving the treatment served as the control group. The cumulative embryo score, fertilization rate, cleavage rate, rate of good quality embryos, implantation rate, clinical pregnancy rate, first trimester and late miscarriage rates, delivery rate, and neonatal malformation rate were recorded and compared between the two groups.

RESULTSBetween the HBsAg-positive and the control groups, the cumulative embryo score (52.8-/+18.7 vs 55.4-/+16.9), insemination rate (66.9% vs 66.1%), cleavage rate (97.6% vs 97.2%), rate of good quality embryos (34.0% vs 37.1%), implantation rate (40.9% vs 34.6%), clinical pregnancy rate (56.9% vs 50%), first trimester miscarriage rate (6.9% vs 5.9%) and late pregnancy miscarriage rate (8.6% vs 4.9%), delivery rate (40.2% vs 43.6%) and neonatal malformation rate (0 vs 0) were all similar (P>0.05;).

CONCLUSIONChronic HBV infection in the husband might not affect the outcome of IVF-ET treatment.

Case-Control Studies ; Embryo Transfer ; Female ; Fertilization in Vitro ; Hepatitis B Surface Antigens ; blood ; Hepatitis B, Chronic ; physiopathology ; Humans ; Male ; Pregnancy ; Pregnancy Outcome

OBJECTIVETo investigate the anti-mutagenic action of the total flavonoids of Astragalus (TFA).

METHODThree groups of concentrations of TFA and one inducer group were used. The anti-mutagenic action of TFA was studied by experiments in vitro and in vivo including miconucleolus assay, semen teraogenesis test and gene mutagenesis of V79/HGPRT.

RESULTTFA did not affect the weight of the mice during the experiment period. The high concentration of TFA could significantly reduce cyclohophamide-induced miconucleolus number and gene mutagenesis of V79/HGPRT compared with inducer group. However, TFA did not make statistic change in the semen teraogenesis induced by mitomycin C.

CONCLUSIONTFA has antimutagenesis effect.

Animals ; Antimutagenic Agents ; isolation & purification ; pharmacology ; Astragalus membranaceus ; chemistry ; Cells, Cultured ; Chromosome Aberrations ; drug effects ; Cricetinae ; Cricetulus ; Flavonoids ; isolation & purification ; pharmacology ; Male ; Mice ; Micronucleus Tests ; Mutagenesis ; drug effects ; Plants, Medicinal ; chemistry ; Testis ; cytology ; drug effects