OBJECTIVETo study the chemical constituents in the active parts of Huoxue Yiqi Tang.

METHODThe silica gel, Sephadex LH-20 were used to isolate the compounds, El, FAB-MS and 1D, 2D-NMR were used to identify the obtained compounds.

RESULT14 compounds were obtained from the prescription.

CONCLUSIONThey were all first time obtained from the prescription. Among them, 7, 3'-dihydroxyl-5'-methoxyisoflavone is new.

Anthraquinones ; chemistry ; isolation & purification ; Drug Combinations ; Drugs, Chinese Herbal ; chemistry ; Emodin ; chemistry ; isolation & purification ; Isoflavones ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry

To determine the optimum conditions of supercritical CO2 extraction of Xiaoyaosan, and establish its fingerprint by gas chromatography-mass spectrometry (GC-MS), the yield of extract were investigated, an orthogonal test was used to quantify the effects of extraction temperature, pressure, CO2 flow rate and time, and fingerprint analysis of different batches of extracts were by GC-MS. The optimal extraction conditions were determined as follows: extraction pressure 20 MPa, extraction temperature 50 degrees C, CO2 flow rate 25 kg x h(-1), extraction time 3 h, and average yield 2.2%. The GC-MS fingerprint was established and 27 common peaks were found, whose contents add up to 81.89% of the total peak area. Among them, 21 compounds were identified, accounting for 53.20% of the total extract. The extraction process is reasonable and favorable for industrial production. The GC-MS method is accurate, reliable, reproducible, and can be used for quality control of supercritical CO2 extract from Xiaoyaosan.
Carbon Dioxide ; chemistry ; Chromatography, Supercritical Fluid ; methods ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Gas Chromatography-Mass Spectrometry ; methods

Objective To analyze the data of gene expression profiles of choriocarcinoma and screen for choriocarcinomarelated genes. Methods Human cDNA expression microarray containing 4 096 genes was used to study the gene expression profiles in specimens of complete hydatidiform moles (n=3) and choriocarcinomas (n=3), with normal placental villi serving as the control group. The candidate genes with similar expression profiles were identified by hierarchical cluster analysis, and their expressions in normal and neoplastic tissues analyzed by electronic Northern analysis and other bioinformatics methods.Three selected genes were analyzed by β-actin semiquantitative reverse transcriptase-PCR to confirm the data. Results A total of 52 coexpressed candidate genes were identified from the gene expression data derived from the choriocarcinoma specimens, of which 19 genes were selected as choriocarcinoma-related genes by further cluster analysis, such as dynamin (L07807), kataninp60 (AF056022), zinc fingerproteinZNF184 (U6656), calmodulin (U12022), carboxypeptidaseM (BC022276), calcineurin-binding protein cabin 1 (NM_012295) and transducin-like enhancer protein TLE1 (M99435).Conclusion The identification of choriocarcinoma-related genes by cluster analysis provides new clues for seeking the key genes associated with the progression and metastasis of choriocarcinoma.

Objective To analyze the data of gene expression profiles of choriocarcinoma and screen for choriocarcinomarelated genes. Methods Human cDNA expression microarray containing 4 096 genes was used to study the gene expression profiles in specimens of complete hydatidiform moles (n=3) and choriocarcinomas (n=3), with normal placental villi serving as the control group. The candidate genes with similar expression profiles were identified by hierarchical cluster analysis, and their expressions in normal and neoplastic tissues analyzed by electronic Northern analysis and other bioinformatics methods.Three selected genes were analyzed by β-actin semiquantitative reverse transcriptase-PCR to confirm the data. Results A total of 52 coexpressed candidate genes were identified from the gene expression data derived from the choriocarcinoma specimens, of which 19 genes were selected as choriocarcinoma-related genes by further cluster analysis, such as dynamin (L07807), kataninp60 (AF056022), zinc fingerproteinZNF184 (U6656), calmodulin (U12022), carboxypeptidaseM (BC022276), calcineurin-binding protein cabin 1 (NM_012295) and transducin-like enhancer protein TLE1 (M99435).Conclusion The identification of choriocarcinoma-related genes by cluster analysis provides new clues for seeking the key genes associated with the progression and metastasis of choriocarcinoma.

The mutations in the dual oxidase 2 (DUOX2) and dual oxidase maturation factor 2 (DUOXA2) genes can cause congenital hypothyroidism (CH). This study reports the pedigree with goitrous congenital hypothyroidism (GCH) due to the coexistence of heterozygous mutations in the DUOX2 and DUOXA2 genes. The two sisters with GCH were diagnosed with CH at neonatal screening and were enrolled in this study. The DUOX2, DUOXA2, and thyroid peroxidase (TPO) genes were considered for genetic defects screening. Family members of the patients and normal controls were also enrolled and evaluated. The two girls harbored compound heterozygous mutations, including a new mutation of c.2654G>T (p.R885L) in the maternal DUOX2 allele and c.738C>G (p.Y246X) in the paternal DUOXA2 allele, that has been previously reported. The germline mutations from the families were consistent with an autosomal recessive inheritance pattern. No mutations in the TPO gene and the controls were observed.
Alleles ; Congenital Hypothyroidism* ; Female ; Germ-Line Mutation ; Humans ; Infant, Newborn ; Inheritance Patterns ; Iodide Peroxidase ; Mass Screening ; Neonatal Screening ; Oxidoreductases ; Pedigree ; Siblings

OBJECTIVETo introduce the newly found gene TIP30/CC3 into a hepatoma cell line PLC/PRF/5 and select the stable expression clones. The growth and cell cycles were studied with the clones stably expressing TIP30/CC3 or anti-TIP30/CC3, and the effects of TIP30/CC3 gene on hepatoma cells were analyzed.

METHODSThe internal expression of TIP30/CC3 protein was detected with Western blot, then TIP30/CC3 or anti-TIP30/CC3 cDNA was subcloned into a constitutive vector pcDNA3 followed by transfection into PLC/PRF/5. Stable expression clones were selected. The cell growth curve was made and cell cycles detected using flow cytometry. To confirm the results in vitro, stable-expressing cells were implanted subcutaneously into nude mice and time of tumor formation recorded and tumor volume measured.

RESULTSPLC-anti-TIP30 grew faster than the others. Three days after transfection, live cells of PLC-anti-TIP30 were 14.0*10(4), in comparison with the control PLC-DNA3 and PLC/PRF/5, the differences were statistically significant. Live cells of PLC-TIP30 were 4.9*10(4), significantly less than the two control groups. Six days after transfection, live cells of PLC-anti-TIP30 were 25.0*10(4), significantly more than the controls PLC-DNA3 and PLC/PRF/5. Live cells of PLC-TIP30 were 12.4*10(4), significantly less than the two control groups. Cell cycle analysis showed that PLC-anti-TIP30 proliferated faster, 22.4% cells were in G0/G1 (gap) phases and 58.6% cells in S (DNA synthesis) phase. The growth of the PLC-anti-TIP30 cell was retarded and many cells were arrested from G1 to S phases. Cells in G0/G1 and S phase were 44.2% and 33.3% respectively. Furthermore, the average time of tumor formation was shorter in anti-TIP30 group and longer in TIP30/CC3 group, and times were 6.0 d (with control groups) and 15.6 d (with control groups) respectively. Tumors in the nude mice grew faster in PLC-anti-TIP30 group and slower in PLC-TIP30 group.

CONCLUSIONTumor suppressor gene TIP30/CC3 can inhibit the proliferation of tumor cells and interfere in its cell cycles. It can be used as a valuable tool for hepatoma biotherapy including gene therapy.

Acetyltransferases ; biosynthesis ; genetics ; Animals ; Carcinoma, Hepatocellular ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Female ; Genes, Tumor Suppressor ; Humans ; Liver Neoplasms ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Transcription Factors ; biosynthesis ; genetics ; Transfection

AIMTo study compounds isolated from Picria fel-terrae.

METHODSThe chemical constituents were separated and purified by column chromatography on silica gel and MCI. Their structures were identified on the basis of spectral data (IR, UV, MS, ID NMR and 2D NMR).

RESULTSA new cucurbitacin, along with a known one, were obtained from the 60% EtOH extract of the whole plant.

CONCLUSIONThe new compound was identified as 11, 24-dioxo-5, 21-diene-cucurbit-3alpha-O-beta-D-xylopyranosyl-16alpha-O-alpha-L-rhamnopyranoside (dehydrobryogenin glycoside). The known one, hexanorcucurbitacin F, was obtained for the first time from Picria fel-terrae.

Molecular Structure ; Plants, Medicinal ; chemistry ; Saponins ; chemistry ; isolation & purification ; Scrophulariaceae ; chemistry ; Steroids ; chemistry ; isolation & purification

China ; Cost of Illness ; Demography ; Female ; Humans ; Maternal Mortality ; Poverty Areas

OBJECTIVETo isolate and identify compounds from Picria fel-tarrae in order to utilize it better.

METHODConstituents from Picria fel-tarrae were isolated by several column chromatography and their structures were elucidated on the basis of chemical and spectral analysis.

RESULTSix compounds, N-benzoylphenylalanyl-L-phenylalaninol acetate (1), 1-hydroxy-7-hydroxymethyl-9,10-anthraquinone (2), 9, 16-dioxo-10,12,14-octadeca-trienoic acid (3), 5,7,4'-trihydroxy-flavone (4), beta-sitosterol (5), and daucosterol (6) were obtained from the fraction with relatively low polarity of EtOH extract of Picria fel-tarrae.

CONCLUSIONCompounds 1-6 were isolated from picria fel-tarrae Lour for the first time, and the 13C-NMR data of compounds 1-3 are provided firstly in the literature.

Apigenin ; chemistry ; isolation & purification ; Dipeptides ; chemistry ; isolation & purification ; Molecular Structure ; Plants, Medicinal ; chemistry ; Scrophulariaceae ; chemistry ; Sitosterols ; chemistry ; isolation & purification

OBJECTIVETo investigate the effect of Compound Qingqin Liquid (CQL) on the expression level of angiotensin II (Ang II) and COX-2 mRNA transcription and protein expression in the renal tissue of rats with uric acid nephropathy.

METHODSSD rats were randomly divided into the blank control group, the model group, the positive drug group, the high, moderate, and low dose CQL group according to number randomization principle. The model was established by gastrogavage of adenine, accompanied with yeast feeding. Distilled water was given by gastrogavage to rats in the blank control group and the model group. Allopurinol at the daily dose of 9.33 mg/kg was given by gastrogavage to rats of the positive control group. CQL at the daily dose of 3.77 g/kg, 1.89 g/kg, and 0.09 g/kg was respectively given by gastrogavage to rats in the high, moderate, and low dose CQL groups. All treatment lasted for 6 weeks. Rats were randomly divided at week 4 (3 in the blank control group, and 6 in the rest groups), and the rest rats were killed at week 6. The renal tissue was extracted. The expression level of Ang II and COX-2 mRNA transcription were detected by RT-PCR. The expression level of Ang II was detected by ELISA. The expression level of COX-2 protein was detected by Western blot and immunohistochemical assay.

RESULTSCompared with the blank control group, except the mRNA expression of Ang II at week 4, the mRNA and protein expression of Ang II and COX-2 obviously increased at week 4 and 6 in the model group (P < 0.01, P < 0.05). The COX-2 protein expression at week 4 was obviously lower in the high and moderate dose CQL groups than in the model group and the low dose CQL group (P < 0.05); the average integral of optical density value was obviously lower in the positive control group than in the model group. Except the mRNA expression of Ang II in the high dose CQL group at week 6, the mRNA and protein expression of Ang II obviously decreased in the positive control group and each dose CQL group (P < 0.01, P < 0.05). Of them, the effects were better in the high and moderate dose CQL groups than in the positive control group and the low dose CQL group (P < 0.05, P < 0.01). Besides, the mRNA expression of COX-2, the average integral of optical density value were obviously lower in the positive control group and each dose CQL group than in the model group (P < 0.05). The protein expression of COX-2 was obviously lower in the high and moderate dose CQL groups than in the model group (P < 0.05). Of them, the mRNA expression of COX-2 was better in the moderate dose CQL group than in the positive control group (P < 0.05); the protein expression of COX-2 was better in the high dose CQL group than in the low dose CQL group (P < 0.05).

CONCLUSIONCQL was capable of lowering the expression level of Ang II, COX-2 mRNA transcription and protein expression, thus suppressing the inflammatory pathological injury of the renal tissue.

Angiotensin II ; metabolism ; Animals ; Cyclooxygenase 2 ; genetics ; metabolism ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Kidney ; metabolism ; Kidney Diseases ; drug therapy ; metabolism ; Male ; RNA, Messenger ; genetics ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Uric Acid