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

To construct plasmid of recombinant protein candidate vaccine of respiratory syncytial virus, express it in E. coli, and to investigate its immunogenicity and protective efficacy. A CD8+ T cell epitope from respiratory syncytial virus (RSV) M2 protein F/M2:81 - 95 and the G:125-225 (G1) gene fragments from RSV-G protein containing B cell epitopes were amplified by PCR method and then inserted into the prokaryotic expression vector pET-DsbA after bonding to a linker. The fusion protein DsbA-G1-Linker-F/M2:81-95 (D-G1LF/M2) was expressed successfully in E. coli BL21 (DE3). The product was proved to be RSV-specific by Western-blot. After purified by affinity chromatography on Ni+ Sepharose and renatured by gradient dialysis. D-G1LF/M2 was used to immune BALB/c mice. D-G1LF/M2 induced high anti-D-G1LF/M2 IgG, anti-RSV IgG and neutralizing antibody titers in serum and lung of BALB/c mice, and elicied RSV-specific CTL responses. The IgG subclass distribution revealed that IgG1/IgG2a ratio was 2.66. Viral titration indicated that D-G1LF/M2 could protect BALB/c mice against RSV challenge in lung.
Animals ; Antibodies, Viral ; blood ; immunology ; Escherichia coli ; genetics ; metabolism ; Humans ; Immunoglobulin G ; blood ; immunology ; Mice ; Mice, Inbred BALB C ; Plasmids ; genetics ; immunology ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology ; Respiratory Syncytial Virus Infections ; prevention & control ; Respiratory Syncytial Virus Vaccines ; biosynthesis ; genetics ; immunology ; Respiratory Syncytial Virus, Human ; genetics ; immunology ; Viral Envelope Proteins ; genetics ; Viral Fusion Proteins ; genetics ; Viral Proteins ; genetics

OBJECTIVEto evaluate the clinical outcomes of transpedicular lumbar wedge resection osteotomy in treating adult idiopathic scoliosis.

METHODStwenty-five adult idiopathic scoliosis patients treated with transpedicular lumbar wedge resection osteotomy from July 2001 to November 2007 were included, among whom 18 were female and 7 were male. Nine of 25 were with double major curve in thoracic and thoracolumbar/lumbar spine, and 16 were with single curve in thoracolumbar/lumbar spine. The average age was 35 years (29 - 48 years) at operation. Osteotomy were performed at T(11), T(12), L(1) or L(2). The motion evoked potential monitoring system and awaking test were used during surgery. The preoperative, postoperative immediately and latest standing posteroanterior and lateral radiographs were reviewed.

RESULTSall patients were operated successfully. The average operation time was 274 min (range, 220 - 380 min) and the average blood loss were 2328 ml (range, 1500 - 5000 ml). The average coronal Cobb angle of all patients in thoracolumbar/lumbar curves was 88° (range, 70° - 121°) before operation, which was corrected to 43° (range, 35° - 70°). The coronal correction rate was 44%. The average kyphosis angle of all in thoracolumbar/lumbar curves was 63° (range, 50° - 90°) before operation, which was corrected to 10° (range, -40° - 21°). The sagittal correction rate was 86%. Nerve root injury occurred in 3 of all patients who complained about postoperative radicular pain. No spine cord injury, delayed paralysis, infection and instrumentation failure were found. With a follow-up of 2 - 4 years, no correction loss or decompensation happened. The back pain existing before operation was relieved in large measure. The cosmetic appearance were all promoted significantly.

CONCLUSIONSthe transpedicular thoracolumbar/lumbar wedge osteotomy is efficient and safe in the correction of adult idiopathic scoliosis. The correction is much better on the sagittal plane than that on the coronal plane.

Adult ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Osteotomy ; methods ; Retrospective Studies ; Scoliosis ; surgery ; Treatment Outcome

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

The aim of this study was to explore the characteristics of Toll-like receptor expression in mesenchymal stem cells derived from bone marrow of healthy donor (BM-MSCs). BM-MSCs were isolated from bone marrow of healthy donor by Ficoll method. Expressions of CD34, CD45, HLA-DR, CD44 and CD71 in BM-MSCs were detected by flow cytometry. CD71 in BM-MSCs was assayed by immunocytochemistry. The adipocyte and osteoblast induction of BM-MSCs were detected by alizarin red stain and oil red stain respectively. TLR 1 - 10 mRNA levels in BM-MSCs were evaluated by semiquantitative RT-PCR. The results showed that expressions of CD34, CD45 and HLA-DR in BM-MSC were negative while the expressions of CD44 and CD71 were positive. CD71 in BM-MSCs was positive. After induced by osteoblast and adipocyte inductor, BM-MSCs were positive for alizarin red staining and oil red staining respectively. All of TLR 1 - 10 mRNA were found in BM-MSCs with high expression levels of TLR2, TLR3, TLR4, TLR7, TLR8, TLR9 and low expression levels of TLR1, TLR5, TLR6, TLR10. In conclusion, different levels of TLR 1 - 10 mRNA were expressed in BM-MSCs of healthy donor.
Bone Marrow Cells ; metabolism ; Cell Differentiation ; Cells, Cultured ; Humans ; Mesenchymal Stromal Cells ; metabolism ; RNA, Messenger ; genetics ; Toll-Like Receptors ; metabolism

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