Objective：This study was designed to evaluate the treatment value of telomerase inhibitors for animal tumor in vivo and investigate radiosensitive effects of telomerase inhibitors. Methods：With orthogonal experiment design, the telomerase inhibitors(azidothymidine AZT 300 mg· kg -1· d -1, lamivudine 150 mg· kg-1· d -1)and radiotherapy (RT, 10 gray in a week, 2 gray/Fx, 5 Fx/wk) were used to treat breast cancers (MA782)xenograft in BALB/c mice for evaluating their influence on tumor size and telomerase expression. The tumor size was measured with a digital vernier scale. Telomerase activity was examined by a PCR-based telomeric repeat amplification protocol (TRAP) coupled with ELISA. Results：The tumor size of mice increased 2.7268 times in untreated and 2.5130 times in lamivudine-administrated, which indicated lamivudine having no obvious effect on tumor volume (P >0.05)； AZT, RT, and AZT combined with RT reduced tumor volume of 20.70％ , 47.43％ , and 85.19％ , respectively, and combined model(AZT＋ RT) was more effective than others used alone(P<0.05); The A value of tumor telomerase was untreated,0.817; AZT,0.453; lamivudine,0.760; RT,0.480; AZT 0.340 combined with RT； and AZT plus lamivudine plus RT,0.166 ; which suggested that AZT, RT and lamivudine could decrease the activity of tumor telomerase (P<0.05), and furthermore, AZT combined with RT have an additive effect (P< 0.05). Conclusion： Both AZT and radiotherapy are effective to treat murine breast cancer through decreasing telomerase activity and reducing tumor size. AZT can increase the radiation sensitivity for murine breast cancer MA782. Telomerase is a new target for anticancer treatment.

Autoimmune Lymphoproliferative Syndrome ; diagnosis ; genetics ; immunology ; therapy ; Biomarkers ; blood ; Child, Preschool ; Humans ; Male ; Mutation ; Prednisolone ; administration & dosage ; therapeutic use ; Ultrasonography, Doppler, Color ; fas Receptor ; genetics

The pharmacokinetics of a long-circulating PEGylated Radix Ophiopogonis polysaccharide (ROP) was investigated in rats following i.v. or s.c. administration at three dose levels (9, 20, 50 mg x kg(-1)). A moderate coupling reaction between the hydroxyl-activated ROP and the amino-terminated mPEG was chosen to produce PEGylate ROP. The grafting degree of the prepared conjugate was 1.03, and the molecular mass of mPEG used was 20 kDa. High-performance gel permeation chromatorgraphy with fluorescein isothiocyanate prelabeling was established to determine levels of the conjugate in plasma. The results showed that the elimination half-life of the conjugate following s.c. administration was basically identical to that after iv administration. An accurate linear correlation was observed between administration doses and areas under the curve of plasma conjugate level vs. time profile, regardless of the administration route. The absolute bioavailability of the conjugate following sc administration was approximately 56%, and the mean in vivo residence time was 52.1 h, increased 2.4 times compared to those of iv administration. In general, linear pharmacokinetics was observed for the conjugate within the dose range studied, and sc should be a promising administration route for the conjugate.

The paper is to report the observation of pharmacokinetic changes of the characteristic ingredients in the combinations of different-dose herbs of Shaoyao-Gancao decoction. After the establishment of HPLC analytical method of plasma effective constituents, rats were orally administered with different-dose herbs of Shaoyao-Gancao decoction. Blood samples at different times after administering these decoctions were collected, and then were analyzed by HPLC fingerprints technology. Pharmacokinetic parameters of characteristic peaks were analyzed by SPSS 15.0 software and DAS 2.0. At last, we looked for the correlation of those pharmacokinetic parameters and the dosage of Gancao. The best dose of Shaoyao-Gancao decoction was at the ratio of 4 to 4, which was consistent with the dose commonly used in ancient times. The absorption of characteristic peaks from Shaoyao-Gancao decoction was related with the dosage of Gancao, and there existed interaction between each characteristic ingredients. There existed the right dose-ratio of Shaoyao and Gancao to get the best effect. The absorptions of effective constitutents were mutual waxing and waning in order to increase biological effects together. It's demonstrated the compatibility connotation at a right dose-ratio of Shaoyao-Gancao decoction through the angle of pharmacokinetics.

The purpose of this study was to evaluate the effect of telomerase inhibitors combined with X-irradiation on bone marrow hematopoiesis in tumor-carrying mice. With an orthogonal experiment design, the telomerase inhibitors [azidothymidine, AZT 300 mg/(kg.day) and lamivudine 150 mg/(kg x day), per os, bid, x 2 weeks] and X-irradiation [total dose 10 Gy (2 Gy x 5) in 1 week] were used to treat BALB/c mice carrying breast cancer MA(782) for evaluating the influence on peripheral blood cells, bone marrow nucleated cells and telomerase activity. Telomerase activity was detected by a PCR-based telomeric repeat amplification protocol (TRAP) coupled with ELISA. The results showed that the number of marrow nucleated cells (x 10(7)/femur) was 2.1875 in untreated group, and 1.7375, 1.7500 and 1.3475 in irradiated, lamivudine and AZT groups, respectively, these suggested that AZT and irradiation could obviously decrease the number of marrow nucleated cells (P< 0.01 or P < 0.05). The peripheral WBC increased 3.7% in untreated mice, and irradiation, lamivudine and AZT reduced 18.09%, 16.19% and 41.00% of WBC, respectively (P < 0.05). Irradiation, lamivudine and AZT showed no obvious effect on RBC and platelet counts (P > 0.05). The telomerase activity (A(450) nm) of marrow cells was 1.498, 1.483, 0.816 and 0.727 in untreated, irradiation, lamivudine and AZT groups, respectively. It is concluded that AZT and lamivudine combined with X-irradiation inhibit bone marrow nucleate cells and the peripheral WBC, manifest inhibitory effect on telomerase activity in murine bone marrow, but have no effect on the peripheral RBC and platelet.
Animals ; Bone Marrow ; drug effects ; radiation effects ; Enzyme Inhibitors ; pharmacology ; Female ; Hematopoiesis ; drug effects ; radiation effects ; Lamivudine ; pharmacology ; Mice ; Mice, Inbred BALB C ; Telomerase ; antagonists & inhibitors ; X-Rays ; Zidovudine ; pharmacology

OBJECTIVE: To elucidate the effects of mammalian sterile 20-like kinase 1 (MST1) gene on tumor necrosis factor (TNF)-α-mediated human umbilical vein endothelial cell (HUVEC) apoptosis. METHODS: Cultured HUVECs were treated with either vehicle or TNF-α (1-100 ng/mL) for 24 hours. Cell apoptosis was measured by TUNEL staining, and MST1 activity was analyzed by Western blot. In order to knock down MST1 expression in HUVECs, cells were transfected with 100 nmol/L MST1 small interference RNA (siRNA) using Lipofectamine 2000 for 24 hours, and the transfection efficiency was analyzed by Western blot. MST1 siRNA-transfected cells were treated with 10 ng/mL TNF-α for an additional 24 hours. Cell apoptosis was measured by TUNEL staining and caspase-3 activity was detected by Western blot. RESULTS: MST1 activity was stimulated in a dose-dependent manner after TNF-α treatment (10, 40, 100 ng/mL) and reached the maximal effect at 100 ng/mL. MST1 activity also paralleled the onset of apoptosis as determined by TUNEL staining (P<0.001). Transfection with MST1 siRNA markedly diminished MST1 gene expression in a dose-dependent manner. MST1 siRNA (100 nmol/L) significantly silenced MST1 gene (P<0.05) and reduced TNF-α-induced endothelial cells apoptosis (P<0.05) by way of inhibiting MST1 gene activation and, accordingly, suppressing caspase-3 activity. CONCLUSION Silencing of MST1 expression by siRNA diminishes TNF-α-mediated human umbilical vein endothelial cell apoptosis by inhibiting the cascade effect of caspase-3.
Apoptosis ; genetics ; Cells, Cultured ; Hepatocyte Growth Factor ; genetics ; metabolism ; Human Umbilical Vein Endothelial Cells ; cytology ; Humans ; Proto-Oncogene Proteins ; genetics ; metabolism ; RNA Interference ; RNA, Messenger ; genetics ; metabolism ; RNA, Small Interfering ; genetics ; Tumor Necrosis Factor-alpha ; pharmacology

The pharmacokinetics of a long-circulating PEGylated Radix Ophiopogonis polysaccharide (ROP) was investigated in rats following i.v. or s.c. administration at three dose levels (9, 20, 50 mg x kg(-1)). A moderate coupling reaction between the hydroxyl-activated ROP and the amino-terminated mPEG was chosen to produce PEGylate ROP. The grafting degree of the prepared conjugate was 1.03, and the molecular mass of mPEG used was 20 kDa. High-performance gel permeation chromatorgraphy with fluorescein isothiocyanate prelabeling was established to determine levels of the conjugate in plasma. The results showed that the elimination half-life of the conjugate following s.c. administration was basically identical to that after iv administration. An accurate linear correlation was observed between administration doses and areas under the curve of plasma conjugate level vs. time profile, regardless of the administration route. The absolute bioavailability of the conjugate following sc administration was approximately 56%, and the mean in vivo residence time was 52.1 h, increased 2.4 times compared to those of iv administration. In general, linear pharmacokinetics was observed for the conjugate within the dose range studied, and sc should be a promising administration route for the conjugate.
Animals ; Area Under Curve ; Biological Availability ; Drug Carriers ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; isolation & purification ; pharmacokinetics ; Half-Life ; Injections, Intravenous ; Injections, Subcutaneous ; Ophiopogon ; chemistry ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Polyethylene Glycols ; chemistry ; Polysaccharides ; administration & dosage ; chemistry ; isolation & purification ; pharmacokinetics ; Rats ; Rats, Sprague-Dawley

This study was aimed to identify the characteristics of childhood myeloid/natural killer cell precursor acute leukemia (M/NKPAL), and to summarize the therapeutical experiences of this rare hematologic malignancy. A child case of M/NKPAL accompanied by CNS leukemia was enrolled in this study, the therapeutic regiments and the results of long time following up were analysed and evaluated. The results showed that the unusual child case of M/NKPAL with CNS infiltration was diagnosed, showing immunophenotype of CD7(+), CD33(+), CD34(+), CD56(+), HLA-DR(+), MPO(-) and negative for other NK cell, T and B cell differentiation antigens; the chromosomal abnormalities were trisomy 8 and deletion of chromosome 12p. The child case was treated with daunorubicin and cytarabine, and achieved complete remission. Then, 5 courses of acute myeloid leukemia-oriented chemotherapy were given as consolidation chemotherapy, all of the 5 courses contained high dose cytarabine. This child case was given 9 times of lumbar puncture and intrathecal injection, besides these, this case was also given cranial radiotherapy with a dose of 36 Gy. After treated with these methods, the child case achieved long-term complete remission. It is concluded that the M/NKPAL is a rare disease with distinctive immunophenotypic characteristics, acute myeloid leukemia-oriented chemotherapy regimen with high dose of cytarabine may be able to induce long-term remission.
Central Nervous System Neoplasms ; therapy ; Child, Preschool ; Consolidation Chemotherapy ; Female ; Humans ; Immunophenotyping ; Karyotyping ; Leukemia, Large Granular Lymphocytic ; therapy ; Leukemia, Myeloid, Acute ; therapy ; Treatment Outcome

In order to explore the diagnosis and therapy of Diamond Blackfan anemia (DBA), the clinical data of 45 cases of DBA admitted in our hospital from February 1994 to July 2011 were analyzed retrospectively. The clinical characteristics, results of laboratory examination, treatment reaction and outcome of disease were investigated. The results indicated that out of 45 children diagnosed as DBA, 14 cases (31.1%) had short stature and physical malformation. All patients had anemia with reticulocytopenia. Thirty-four patients (75.6%) had mean corpuscular volume. Eleven patients (24.4%) had macrocytic anemia. Bone marrow examination showed a marked erythroid hypoplasia in all patients. Out of 29 cases tested for fetal hemoglobin (HbF), 13 cases (44.8%) had high level of HbF. Erythroid colony-forming unit of bone marrow was tested in 25 patients, among them 12 patients (48%) showed normal plasia, 13 (52%) showed hypoplasia. The erythropoietin (EPO) levels of 17 patients were elevated. Karyotypes were examined in 28 patients, and showed all normal. The treatment was based on corticosteroids and Cyclosporine A. Thirty patients had good response to corticosteroid therapy, and 10 of them obtained a sustained corticosteroid-induced remission. Twenty cases discontinued corticosteroid therapy after remission, as a result, 15 cases (75%) relapsed, moreover all the relapsed cases still had good response to corticosteroid. Two relapsed patients suffered from aplastic anemia, one of them died of therapy failure. Six patients were unresponsive to corticosteroid, 1 of which achieved remission with cyclosporine A and the others continued to receive regular transfusions. 3 patients received iron chelation therapy. It is concluded that the clinical characteristics, complete blood count, bone marrow smear, HbF level and EPO level are useful to make a diagnosis of DBA. Most patients have a good response to corticosteroid therapy, but relapse rate is high when drug was discontinued. Patients unresponsive to corticosteroid should receive regular transfusions and chelation therapy.
Anemia, Diamond-Blackfan ; diagnosis ; therapy ; Bone Marrow Examination ; Child ; Child, Preschool ; Erythroid Precursor Cells ; Female ; Humans ; Infant ; Male ; Retrospective Studies

OBJECTIVETo study the therapeutic effects of azithromycin in treatment of congenital toxoplasmosis in children.

METHODSDefinite diagnosis of congenital toxoplasmosis was made on the basis of clinical manifestation combined with one or more positive results of the following laboratory tests and excluded other congenital infectious diseases: toxoplasma DNA (TOX-DNA), circulating toxoplasma antigen (TOX-CAG), and toxoplasma IgM antibody (TOX-IgM). All the patients were given oral azithromycin 10 mg/(kg.d) for 6 days followed by 8 days without medication (one course of treatment), and the regimen was persisted for 2 months and then another 2-month treatment was given at a 1-month interval. The authors continued to provide further treatment according to the state of the illness at one month interval. The patients received 2 to 8 (average 5) courses of treatment. The patients were followed-up for 2.5 to 5 (average 4) years.

RESULTSThe treatment was effective in all the patients and the patient's condition was improved. The authors repeated in 12 cases the four tests for toxoplasma (TOX-DNA, TOX-CAG, TOX-IgM, and TOX-IgG) 9 months to one and a half years after treatment. In 10 cases all these tests showed negative results, in 2 cases TOX-IgG was positive and in the other 4 cases symptoms disappeared.

CONCLUSIONThe results of the study showed that oral azithromycin had significant therapeutic effects with little side effect and was well tolerated. Azithromycin may become an alternative therapy in treatment of congenital Toxoplasma gondii infection in children.

Anti-Bacterial Agents ; administration & dosage ; therapeutic use ; Azithromycin ; administration & dosage ; therapeutic use ; Female ; Follow-Up Studies ; Humans ; Infant ; Infant, Newborn ; Male ; Prognosis ; Toxoplasmosis, Congenital ; diagnosis ; drug therapy ; Treatment Outcome