No abstract available.
Galactitol* ; Galactosemias* ; Humans ; Infant* ; Mass Screening*

BACKGROUND: To access the accuracy and clinical usefulness of microplate identification (ID) system for the identification of Enterobacteriaceae, we compared microplate ID system with API 20E(bioMerieux, Etoile, France). METHODS: Ninety-two cultures of Enterobacteriaceae and one isolate of Aeromonas species were simultaneously identified by microplate ID system and the API 20E. Twenty biochemical tests used in microplate ID system were lactose, sucrose, and H2S in Kligler's iron agar media; indole, sucrose, raffinose, arabinose, trehalose, adonitol, dulcitol, sorbitol, cellibiose, methy-red, phenylalanine deaminase, ornithine decarboxylase, lysine decarboxylase, arginine dihydrolase, urease, and citrate in microplate; and oxidase test. The identification was obtained by considering percent likelihood(% ID), modal frequency and ID score method. RESULTS: Among the 92 cultures of Enterobacteriaceae and one isolate of Aeromonas species, agreement rate of identification according to the % ID between microplate ID system and API 20E were 90.3% to the species level and 97.8% to the genus level. CONCLUSIONS: For the identification of clinical Enterobacteriaceae isolates, the microplate ID system compares favorably with API 20E in identification accuracy and have the advantage of costsaving and easy to use.
Aeromonas ; Agar ; Arabinose ; Arginine ; Citric Acid ; Enterobacteriaceae* ; Galactitol ; Iron ; Lactose ; Lysine ; Ornithine Decarboxylase ; Oxidoreductases ; Phenylalanine ; Raffinose ; Ribitol ; Sorbitol ; Sucrose ; Trehalose ; Urease

OBJECTIVETo establish the processing method of Cistanche tubulosa decoction pieces.

METHODThe orthogonal test of four factors and three levels was used to optimize the main factors in the process of fresh C. tubulosa decoction pieces processing, including the thickness, temperature, and the time for inactivation of the enzyme in the plant. The result showed that the optimized condition was that fresh C. tubulosa was cut into 4 mm thickness, and heated at 70 degrees C for inactivation the enzyme in the plant for 6 min. Moreover, the optimized method was compared with the method of insolation and traditional dried method.

RESULTThe content of echinacoside in the C. tubulosa decoction piece by the optimized method was 7.3 times of that dried by insolation, and 12.8 times of that by traditional dry method; the content of verbascoside was 6. 5 and 14. 9 times of that dried by insolation and by traditional dry method, respectively; the content of galactitol was 7.1 and 13.2 times of that dried by insolation and by traditional dry method, respectively.

CONCLUSIONThe quality of C. tubulosa decoction pieces could be improved by this method, and its crud drug could be saved, which would protect the source of the mild Herba Cistanche, and produced the better economic and ecological benefits.

Cistanche ; chemistry ; Desiccation ; methods ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; standards ; Galactitol ; analysis ; Glucosides ; analysis ; Glycosides ; analysis ; Hot Temperature ; Phenols ; analysis ; Plants, Medicinal ; chemistry ; Quality Control ; Technology, Pharmaceutical ; methods

OBJECTIVETo explore the expression of multidrug resistance gene 1 ( MDR1), glutathione-S-transferases-pi (GST-pi) in osteosarcoma and soft tissue sarcoma tissues from 34 patients and their correlation with chemotherapy resistance.

METHODSMDR1 and GST-pi expressions were analyzed by real-time fluorescence quantitative polymerase chain reaction (FQ-PCR) and flow cytometry (FCM) at mRNA and protein levels, respectively. Chemotherapy sensitivity on adriamycin, cisplatinum, fluorouracil, mitomycin C, dacarbazine, vincristine, methotrexate in tumor tissues were detected by MTT assay.

RESULTSThe nonsensitive rates on adriamycin, cisplatinum, fluorouracil, mitomycin C, dacarbazine, vincristine, methotrexate in tumor tissues were 41.18%, 17.7%, 47.1%, 50.0%, 76.5%, 61.8% and 52.9%, respectively. The expression of P-glycoprotein (P-gp) and GST-pi in tumor tissues was 1.54 and 2.58 (relative fluorescence intensity). Chi2 analysis showed that there was a positive correlation between P-gp expression and drug resistance on ADM, GST-pi expression and resistance on ADM, DDP and MMC (P < 0.05). There was not seen obvious correlation between expression of MDR1, GST-pi and age, gender, pathological type, tumor size in osteosarcoma and soft tissue sarcoma patients (P > 0.05). The expression of GST-pi was increased in patients receiving preoperative chemotherapy. The rate of postoperative recurrence was higher in patients with higher GST-pi expression level than those with lower GST-pi expression level before operation (P < 0.05).

CONCLUSIONIndividual differences exist in chemotherapy sensitivity and expression of MDR1 and GST-pi in osteosarcoma and soft tissue sarcomas patients. Chemotherapy can induce up-regulation of GST-pi protein expression. Primary high expression of GST-pi is the main mechanism of resistance of osteosarcoma and soft tissue sarcomas to chemotherapy and is related to poor prognosis.

ATP-Binding Cassette, Sub-Family B, Member 1 ; biosynthesis ; genetics ; Adolescent ; Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Bone Neoplasms ; drug therapy ; genetics ; metabolism ; Child ; Cisplatin ; therapeutic use ; Doxorubicin ; therapeutic use ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; Female ; Flow Cytometry ; Follow-Up Studies ; Glutathione S-Transferase pi ; biosynthesis ; genetics ; Humans ; Male ; Middle Aged ; Mitolactol ; therapeutic use ; Mitomycins ; therapeutic use ; Osteosarcoma ; drug therapy ; genetics ; metabolism ; Prognosis ; RNA, Messenger ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Sarcoma ; drug therapy ; genetics ; metabolism

BACKGROUNDBoth survivin and lung resistance related protein (LRP) are related to the chemoresistances in hepatocellular carcinoma (HCC). But the relationship between survivin and LRP is indefinite. The aim of this study was to investigate the effects of down-regulation of survivin on LRP expressions and the reversal of chemoresistances in HCC both in vitro and in vivo.

METHODSThe expressions of survivin were detected by RT-PCR and Western blotting in HCC cell line SMMC-7721 and SMMC-7721/ADM. The sensitivities of these two cell lines to ADM were evaluated by MTT assays. SiRNA which targeted survivin was transfected into SMMC-7721/ADM cells, then the sensitivity of SMMC-7721/ADM cells to ADM and the expressions of survivin and LRP were detected respectively. SMMC-7721/ADM cells were transplanted subcutaneously into nude mice to establish xenograft tumors. Antitumor activities of RNA interference (RNAi) targeting survivin, various doses of ADM and combination therapies were observed respectively. Possible toxicities were evaluated. LRP expression changes were tested. Student's t test was used for evaluating statistical significance.

RESULTSThe expressions of survivin in SMMC-7721/ADM cell line showed significant elevation compared to those in SMMC-7721 cell line (P < 0.05). Positive siRNA down-regulated the expressions of survivin significantly (P < 0.05). SiRNA targeting survivin could sensitize SMMC-7721/ADM cells to ADM and down-regulate the expressions of LRP significantly (P < 0.05). Growths of the tumors were significantly inhibited in positive siRNA group as compared with those in the control group from the 8th day (P < 0.05). Combination therapies caused significant tumor inhibitions compared with tumors of nude mice in the other three groups respectively (P < 0.05). No toxicities were found in nude mice treated by siRNA and combination therapies. The expressions of LRP were markedly reduced in tumors treated with siRNA targeting survivin (P < 0.05).

CONCLUSIONSDown regulation of survivin gene by RNAi can increase chemosensitivity of HCC both in vitro and in vivo. The reversal of drug resistance may be reduced through the inhibitions of LRP.

Animals ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Blotting, Western ; Carcinoma, Hepatocellular ; drug therapy ; genetics ; metabolism ; Cell Line, Tumor ; Doxorubicin ; therapeutic use ; Drug Resistance, Neoplasm ; Humans ; Inhibitor of Apoptosis Proteins ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Microtubule-Associated Proteins ; genetics ; metabolism ; Mitolactol ; therapeutic use ; Mitomycins ; therapeutic use ; RNA Interference ; physiology ; RNA, Small Interfering ; genetics ; physiology ; Reverse Transcriptase Polymerase Chain Reaction ; Vault Ribonucleoprotein Particles ; genetics ; metabolism ; Xenograft Model Antitumor Assays

AIMTo investigate the apoptosis induced by diacetyldianhydrogalactitol (DADAG) and its mechanism in human HL-60 leukemia cells.

METHODSInhibition of proliferation was measured by MTT assay. DADAG-induced apoptosis in HL-60 cells was observed by electron microscopy, flow cytometry and DNA fragmentation assay. The levels of Bcl-2 family proteins were detected by Western blotting. Caspase-3 activity was determined by ApoAlert CPP32 colorimetric assay kit.

RESULTSDADAG exhibited potent antiproliferative activity and induced apoptosis in HL-60 cells. After treatment with DADAG 8 micrograms.mL-1 for various times, the Bcl-XL protein level decreased in a time-dependent manner, while the Bad protein level was upregulated. The caspase-3 activity increased markedly after treatment with DADAG for 24 h. The apoptotic signals were suppressed by z-VAD.fmk (a general inhibitor of caspases), whereas z-DEVD.fmk, a selective inhibitor of caspase-3, only induced partial reversion of the apoptotic effects.

CONCLUSIONDADAG-induced apoptosis in HL-60 cells required caspase-3 activation and caspase-3 activation was related with Bcl-2 family members.

Antineoplastic Agents ; pharmacology ; Apoptosis ; Caspase 3 ; Caspases ; metabolism ; Cell Division ; drug effects ; Dianhydrogalactitol ; analogs & derivatives ; pharmacology ; HL-60 Cells ; Humans ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; bcl-X Protein