Chemotherapy initiation with single-course methotrexate alone or combined with dactinomycin versus multi-course methotrexate for low-risk gestational trophoblastic neoplasia: a multi-centric randomized clinical trial / 医学前沿

We aimed to evaluate the effectiveness and safety of single-course initial regimens in patients with low-risk gestational trophoblastic neoplasia (GTN). In this trial (NCT01823315), 276 patients were analyzed. Patients were allocated to three initiated regimens: single-course methotrexate (MTX), single-course MTX + dactinomycin (ACTD), and multi-course MTX (control arm). The primary endpoint was the complete remission (CR) rate by initial drug(s). The primary CR rate was 64.4% with multi-course MTX in the control arm. For the single-course MTX arm, the CR rate was 35.8% by one course; it increased to 59.3% after subsequent multi-course MTX, with non-inferiority to the control (difference -5.1%,95% confidence interval (CI) -19.4% to 9.2%, P = 0.014). After further treatment with multi-course ACTD, the CR rate (93.3%) was similar to that of the control (95.2%, P = 0.577). For the single-course MTX + ACTD arm, the CR rate was 46.7% by one course, which increased to 89.1% after subsequent multi-course, with non-inferiority (difference 24.7%, 95% CI 12.8%-36.6%, P < 0.001) to the control. It was similar to the CR rate by MTX and further ACTD in the control arm (89.1% vs. 95.2%, P =0.135). Four patients experienced recurrence, with no death, during the 2-year follow-up. We demonstrated that chemotherapy initiation with single-course MTX may be an alternative regimen for patients with low-risk GTN.

Medication Analysis and Care for Acute Renal Failure with Epileptic Seizure Caused by Excretion Delay of HD- MTX by Clinical Pharmacists / 中国药房

OBJECTIVE：To investi gate the role of clinical pharmacists in the treatment of delayed excretion of acute renal failure （ARF） with epileptic seizure caused by HD-MTX in a patient ，and to provide reference for rational drug use and pharmaceutical care in such type of patients. METHODS ：A patient with diffuse large B-cell lymphoma was given HD-MTX for chemotherapy，and ARF caused by delayed methotrexate excretion occurred on the second day after methotrexate administration. Clinical physicians adjusted the rescue dose and frequency of calcium folinate but the effect was poor. Clinical pharmacists analyzed the causes of delayed methotrexate excretion by reviewing literature and combining with the patient ’s condition. It was suggested to monitor the blood concentration of methotrexate ，strengthen alkalization and hydration ，increase the volume of intravenous sodium bicarbonate from 125 mL to 250 mL，take Sodium bicarbonate tablets orally ，and monitor the pH value of urine （pH value of urine maintained above 7）. In addition ，the pharmacist told the patient to drink water as much as possible to ensure the daily urine output reached 3 000 to 4 000 mL. The blood concentration of methotrexate was 16.14 μmol/L 44 h after administration ，which proved to be excretion delay. The patient had epileptic seizure on the 13th day after methotrexate medication. The physician gave sodium valproate 0.8 g intravenously to control epilepsy. The clinical pharmacist conducted pharmaceutical care for the patient ，and found that the compliance of the patient taking Sodium bicarbonate tablets and Sodium valproate tablets orally was not good ，so medication education and pharmaceutical care were conducted ，then the patient accepted and took the drugs on time. RESULTS ： The physician adopted the suggestions of the pharmacist to monitor the blood concentration of methotrexate and performed symptomatic treatment. The urine volume of the patient increased ，the edema was reduced ，serum creatinine gradually returned to normal，and renal function recovered gradually ；the symptoms of epilepsy was controlled. CONCLUSIONS ：In the treatment process of ARF complicated with epileptic seizure caused by excretion delay of HD-MTX ，the clinical pharmacist assisted physician to improve the treatment plan and conducted pharmaceutical care and medication education for the patient ，therefore ensure the safe and rational use of drugs .

The clinical observation of Compound Xuanju Capsules Combined with methotrexate treating rheumatoid arthritis / 中成药

AIM To evaluate the clinical effects of Compound Xuanju Capsules (CXC,Polyrhachis vicina Roger,Epimedii Folium,Lycii Fructus,Cnidii Fructus) combined with methotrexate (MTX) on patients with moderate or severe rheumatoid arthritis (RA).METHODS One hundred and eighty RA patients were randomized into 3 groups:CXC group,MTX group and combined group (dosed with both CXC and MTX) for a 48-week intervention.The clinical observation on the changes of the signs and symptoms,erythrocyte sedimentation rate (ESR),C-reactive protein (CRP),health assessment questionnaire (HAQ),visual analogue scale (VAS) and disease active score (DAS) 28 were performed before and after each treatment.ACR20,American College of Rheumatology 20％ improvement criteria was taken as the primary end point,and ACR50 and ACR70 as the secondary end points to standardize the patients' response measurement,and all adverse reactions were recorded as well.RESULTS At the time point right after the 12th week,the ACR20 response rate of the combined group (41.5％) was significantly higher than that of the CXC group (19.6％,P ＜ 0.05) and the MTX group (24.1％,P＜0.05).The respective ACR50 (2.1％) and ACR70 (20.8％) response rate of the combined group were significantly higher than those of CXC group and MTX group (P ＜ 0.05).At the time point right after the 24th week,the combined group still demonstrated its significantly higher ACR20 response rate (81.1％) to the CXC group (30.4％,P ＜ 0.05) and the MTX group (68.5％,P ＜ 0.05).The similar superiority in ACRS0 (60.4％) and ACR70 (54.7％) response rate of the combined group to the CXC group and the MTX group were found (P ＜0.05).After the 48th week,the combined group displayed its significantly higher ACR70 response rate (75.7％) to CXC group and MTX group (P ＜ 0.05).Given the reduction of DAS28,HAQ and VAS from the12thweek's5.26±0.83),(22.2±10.3),(6.0±0.4) to the 24thweek's (4.21 ±0.91),(17.1±10.3),(2.4±2.2),andthe48thweek's (2.19±0.56),(10.4±5.0),(0.8±0.9),the combined group's more outstanding performance compared to CXC group and MTX group started right after the 24th week (P ＜ 0.05).Generally,no difference in adverse events was detected between the combined group and MTX group (P ＞ 0.05).CONCLUSION The combined use of CXC and MTX can be an effective and safe treatment for moderate and severe rheumatoid arthritis.

Correlation between Acute Drug-induced Liver and Kidney Injury and HD-MTX Blood Concentration in Patients with Osteosarcoma / 中国药师

Objective: To analyze the correlation between HD-MTX blood concentration and acute drug-induced liver and kidney injury in the patients with osteosarcoma, and investigate the significance of HD-MTX concentration in the monitoring of liver and kidney toxicity. Methods: A total of 56 osteosarcoma patients treated with HD-MTX were selected, and after HD-MTX treatment, the blood concentration of MTX was detected by an HPLC-UV method in 48 h and 72 h after the administration. The liver and kidney function were measured at the same time. The correlation between the different concentrations of MTX and the change of liver and kidney func-tion was analyzed. Results: All the patients were monitored MTX blood concentration at different time points. After the 48-hour HD-MTX treatment, 4 patients (7. 14% ) were with acute drug-induced liver injury and 13 patients (23. 21% ) showed drug-induced kid-ney injury. The average C48hof liver injury was (2.90 ±0.78) μmol·L-1, and the average C48hof kidney injury was (1.65 ±1.12) μmol·L-1. After the 72-hour HD-MTX treatment, 7 patients ( 12. 50% ) were with drug-induced liver injury and 16 patients (28.57%) showed drug-induced kidney injury. The average C72hof liver injury was (0.30 ±0.17) μmol·L-1, while the average C48hof kidney injury was (0. 29 ± 0. 29) μmol·L-1. The function indices of liver ( ALT, ALP and TBIL) and kidney ( SCr) were significantly higher than those in the normal group (P<0. 05), and the blood concentration of MTX was partly significantly correlated with those indicators. Conclusion: There is a certain correlation between MTX induced injury and the blood drug concentration at par-ticular points, and C48hmay be more valuable to predict drug-induced liver and kidney injury.

Successful Treatment of Jesnner's Lymphocytic Infiltration of the Skin with Methotrexate / 대한피부과학회지

No abstract available.

Deoxyadenosine antagonizes MTX-induced suppression of inflammatory bone destruction / 实用口腔医学杂志

Objective:To study the effects of deoxyadenosine(dAdo) on methotrexate (MTX) induced suppression of inflammatory bone destruction.Methods:The culture system of whole bone marrow cells (WBMCs) was utilized to evaluate osteoclastogenesis.TRAP staining and μCT analysis were utilized to evaluate osteoclastogenesis and bone destruction in adjuvant arthritis rats.Results:In the bone marrow culture system,MTX-induced suppression of osteoclastogenesis was abrogated by the addition of dAdo.dAdo canceled MTX-induced suppression of osteoclastogenesis in the rats with arthritis,and significantly abolished the therapeutic effects of MTX on inflammatory bone destruction in the rats.Conclusion:The accumulation of dAdo may be one cause of the losing effectiveness of MTX in bone destruction.

Efecto antimetastásico de celecoxib/PLGA en un modelo murino de adenocarcinoma mamario TA3-MTX-R / Antimetastatic effect of celecoxib/PLGA in a TA3-MTX-R murine breast adenocarcinoma model

La metástasis es el proceso de propagación de un foco cancerígeno a un órgano distinto de aquel en que se inició; ocurriendo generalmente por vía sanguínea o linfática. La localización más frecuentes de las metástasis son los órganos más irrigados como el cerebro, pulmones, hígado, huesos y glándulas suprarrenales. El objetivo fue analizar el patrón de metástasis hepática de tumor TA3-MTX-R, luego de la aplicación del antiangiogénico Celecoxib microencapsulado en PLGA en ratones, así como la disminución de áreas metastásicas a nivel lobulillar. Se utilizó un modelo de tumor experimental, inducido por células TA3-MTX-R, en 18 ratones, separados en 3 grupos de 6 animales, los cuales fueron tratados con dos presentaciones de Celecoxib en administración intramuscular (Grupo 1, Control: TA3-MTX-R; Grupo 2: TA3-MTX-R+Cx y Grupo 3: TA3-MTX-R+Cx/PLGA). Los ratones fueron sacrificados y procesados histológicamente para ser teñidos con H&E y Tricrómico de Arteta. El estudio reveló que el higado muestra una marcada heterogeneidad, y un patrón de metástasis perivascular y neovascularización central y periférica. Además, Celecoxib redujo significativamente la invasión tumoral en el hígado (p<0,0001). Los resultados son similares a descripciones parciales realizadas previamente y son comparables a otras líneas tumorales. Creemos que la vía de administración del fármaco es crítica para la interpretación de los resultados. Los hallazgos son importantes para la discusión de otras investigaciones en donde Celecoxib es usado como un fármaco antiangiogénico.

Metastasis is the propagation process of a cancerous focus to an organ other than that in which it started; usually occurring through blood or lymphatic route. The most common sites of metastases are the organs most irrigated such as the brain, lungs, liver, bones and adrenal glands. The objective was to analyze the pattern of liver tumor metastasis TA3-MTX-R, after application of antiangiogenic Celecoxib microencapsulated in PLGA in mice and decreased metastatic to lobular level areas. An experimental model of tumor induced TA3-MTX-R cells was used, 18 mice divided into 3 groups of 6 animals, which were treated with two presentations Celecoxib intramuscular (Group 1, control was used -R; Group 1: TA3-MTX-R+Cx and Group 3: TA3-MTX-R+Cx/PLGA). The mice were sacrificed and processed histologically to be stained with H&E and Arteta trichrome. The study revealed that the liver shows a marked heterogeneity, and a pattern of perivascular metastasis and central and peripheral neovascularization. Furthermore, Celecoxib significantly reduced tumor invasion in the liver (p <0.0001). The results are similar to partial descriptions made previously and are comparable to other tumor lines. It is believed that the route of administration of the drug is critical for the interpretation of the results. These are important for the discussion of other investigations in which Celecoxib is used as an antiangiogenic drug.

Deoxyadenosine antagonized methotrexate-induced suppression of osteoclastogenesis / 实用口腔医学杂志

Objective:To study the mechanism of the effectiveness loss of methotrexate(MTX)in the treatment of rheumatoid arthri-tis.Methods:The culture system of rat whole bone marrow cells(WBMCs)and tartrateresistant acid phosphatase(TRAP)staining were utilized to evaluate osteoclastogenesis.The mRNA expression of osteoclastogenesis factors in the WBMCs culture system was examined by semi-quantitative RT-PCR.Results:Deoxyadenosine(dAdo)decreased MTX-induced suppression of osteoclastogenesis.The recov-ery effect of dAdo on MTX was partially prevented by caffeine.MTX significantly reduced mRNA expression of receptor activator of nu-clear factor kappa-B ligand(RANKL),dAdo partially recovered RANKL mRNA expression and inhibited osteoprotegerin(OPG)expres-sion.Conclusion:The accumulation of dAdo may induce the effectiveness loss of methotrexate in rheumatoid arthritis treatment.Com-bination of MTX and caffeine can be a potential therapeutic strategy.

Effects of mucus layer on trimethyl chitosan nanoparticles with different degree of quaternization / 中国药学杂志

OBJECTIVE: To study the effects of degree of quaternization(DQ) and mucus on cellular uptake and transport of trimethyl chitosan (TMC) nanoparticles.

Monoterapia biológica en pacientes con artritis reumatoidea en Argentina / Biological monotherapy in patients with rheumatoid arthritis in Argentina

Introducción: La utilización de agentes biológicos para el tratamiento de la Artritis Reumatoidea (AR) es habitualmente usada en aquellos pacientes con enfermedad activa que no hayan respondido al tratamiento con drogas modificadoras de la Artritis Reumatoidea convencionales (DMARD, por sus siglas en inglés) o que hayan presentado intolerancia a las mismas. Al estado actual de la evidencia, la terapia combinada de agentes biológicos más un DMARD convencional (principalmente metotrexato) constituye el estándar de tratamiento. Sin embargo existen algunos escenarios como la intolerancia, la falta de adherencia y la aparición de eventos adversos a las DMARDs convencionales donde la monoterapia biológica emerge como una opción terapéutica válida. Según los distintos registros a nivel internacional, la frecuencia de utilización de agentes biológicos en monoterapia oscila entre 12 a 39%. Debido a la ausencia de estos datos a nivel local decidimos realizar este estudio para conocer el porcentaje de pacientes que se encuentran en monoterapia biológica y analizar las causas que llevaron a este tipo de tratamiento. Materiales y métodos: Estudio de tipo corte transversal donde se invitó a participar a diferentes centros reumatológicos distribuidos a lo largo de Argentina. Cada centro revisó las historias clínicas de los últimos 30 a 50 pacientes consecutivos vistos con AR, mayores de 18 años, que habían presentado inadecuada respuesta al tratamiento con DMARDs y que estaban bajo tratamiento biológico. Se completaba una ficha por cada paciente incluido, registrando datos demográficos, de la enfermedad y tratamientos previos. Resultados: Se incluyeron 32 centros y se evaluaron 1148 historias clínicas de pacientes con AR durante el mes de octubre y noviembre del 2012. Un 21,4% (246) de los pacientes al momento del estudio se encontraba bajo tratamiento biológico en monoterapia...

Introduction: The use of biological agents for the treatment of rheumatoid arthritis (RA) is commonly used in patients with active disease who have not responded to treatment with conventional rheumatoid arthritis-modifying drugs (DMARDs) or Who have presented intolerance to them. At the present state of evidence, combined therapy of biological agents plus conventional DMARD (mainly methotrexate) is the standard of treatment. However, there are some scenarios such as intolerance, lack of adherence and the appearance of adverse events to conventional DMARDs where biological monotherapy emerges as a valid therapeutic option. According to different international registries, the frequency of use of biological agents in monotherapy ranges from 12 to 39%. Due to the absence of these data at the local level we decided to carry out this study to know the percentage of patients who are in biological monotherapy and to analyze the causes that led to this type of treatment. Materials and methods: A cross-sectional study where different rheumatologic centers throughout Argentina were invited to participate. Each center reviewed the medical records of the last 30 to 50 consecutive patients seen with RA, older than 18 years, who had inadequate response to treatment with DMARDs and who were under biological treatment. One card was completed for each patient included, recording demographic, disease and previous treatment data. Results: Thirty-two centers were included and 1148 clinical records of patients with RA were evaluated during October and November 2012. A total of 244 patients (246) at the time of the study were under monotherapy...

Grade 4 oral mucositis and prolonged pancytopenia with high dose intravenous methotrexate in a patient of philadelphia positive acute lymphoblastic leukemia.

Antimetabolite drugs are rarely associated with grade 3 or 4 mucositis. However specific side effects such as bone marrow suppression, pancytopenia, hospitalization, high cost limit the use of intravenous methotrexate. Here we report a case of 25-year-old girl patient with Acute Lymphoblastic Leukemia (ALL) treated with 24 hour methotrexate in her continuous phase of chemotherapy. After 12 hours of completing the treatment she developed oral mucosal pain & she was not able to speak. After a week, her White Blood Cell (WBC) count was 600/cumm with fever and she could not speak and eat or drink. So she was hospitalized for few days till she can drink something. We report high grade mucositis with methotrexate. Hematologists should be aware of this possible side effect to undertake early intervention.

Effect of 5-FU and MTX on the Expression of Drug-resistance Related Cancer Stem Cell Markers in Non-small Cell Lung Cancer Cells

Cancer stem cells (CSCs) are often characterized by the elevated expression of drug-resistance related stem-cell surface markers, such as CD133 and ABCG2. Recently, we reported that CSCs have a high level of expression of the IL-6 receptor (IL-6R). The purpose of this study was to investigate the effect of anticancer drugs on the expression of the drug resistance-related cancer stem cell markers, ABCG2, IL-6R, and CD133 in non-small cell lung cancer (NSCLC) cell lines. A549, H460, and H23 NSCLC cell lines were treated with the anticancer drugs 5-fluorouracil (5-FU; 25 microg/ml) and methotrexate (MTX; 50 microg/ml), and the expression of putative CSC markers was analyzed by fluorescent activated cell sorter (FACS) and the gene expression level of abcg2, il-6r and cd133 by reverse transcriptasepolymerase chain reaction (RT-PCR). We found that the fraction of ABCG2-positive(+) cells was significantly increased by treatment with both 5-FU and MTX in NSCLC cells, and the elevation of abcg2, il-6r and cd133 expressions in response to these drugs was also confirmed using RT-PCR. Also, the number of IL-6R(+) cells was increased by MTX in the 3 cell lines mentioned and increased by 5-FU in the H460 cell line. The number of CD133(+) cells was also significantly increased by both 5-FU and MTX treatment in all of the cell lines tested. These results indicate that 5-FU and MTX considerably enhance the expression of drug-resistance related CSC markers in NSCLC cell lines. Thus, we suggest that antimetabolite cancer drugs, such as 5-FU and MTX, can lead to the propagation of CSCs through altering the expression of CSC markers.

Profiling single nucleotide polymorphisms (SNPs) across intracellular folate metabolic pathway in healthy Indians.

Background & objectives: Many pharmacologically-relevant polymorphisms show variability among different populations. Though limited, data from Caucasian subjects have reported several single nucleotide polymorphism (SNPs) in folate biosynthetic pathway. These SNPs may be subjected to racial and ethnic differences. We carried out a study to determine the allelic frequencies of these SNPs in an Indian ethnic population. Methods: Whole blood samples were withdrawn from 144 unrelated healthy subjects from west India. DNA was extracted and genotyping was performed using PCR-RFLP and Real-time Taqman allelic discrimination for 12 polymorphisms in 9 genes of folate-methotrexate (MTX) metabolism. Results: Allele frequencies were obtained for MTHFR 677T (10%) and 1298 C (30%), TS 3UTR 0bp (46%), MDR1 3435T and 1236T (62%), RFC1 80A (57%), GGH 401T (61%), MS 2756G (34%), ATIC 347G (52%) and SHMT1 1420T (80%) in healthy subjects (frequency of underlined SNPs were different from published study data of European and African populations). Interpretation & conclusions: The current study describes the distribution of folate biosynthetic pathway SNPs in healthy Indians and validates the previous finding of differences due to race and ethnicity. Our results pave way to study the pharmacogenomics of MTX in the Indian population.

Hepatotoxicity of Combination Treatment with Leflunomide and Methotrexate in RA Patients / 대한류마티스학회지

OBJECTIVE: Leflunomide is the newest disease-modifying anti-rheumatic drug (DMARD) that is known to have an equivalent clinical efficacy and tolerability to methotrexate (MTX). Previous studies reported that a co-treatment with MTX and leflunomide can induce additive clinical improvements in RA patients. However, a previous study also demonstrated a reversible elevation of the transaminase levels in up to 63% of patients administered a combination treatment of leflunomide and MTX. This study examined the hepatotoxicity of a combination treatment of MTX and leflunomide. METHODS: From March, 2004, to February, 2006, 203 patients who had been treated in 3 rheumatology clinics, Goyang city, South Korea, were reviewed retrospectively. The data showed that 38.92% of patients scored higher than grade 1 hepatotoxicity and 6.90% of patients scored higher than grade 2 according to the NCI/NIH (National Cancer Institute/National Institutes of Health) Common Toxicity Criteria. RESULTS: The median onset time of hepatotoxicity was 5.91 months after treatment. Leflunomide administration was stopped in 39 patients due to several adverse reactions. Among the 39 patients, hepatotoxicity was observed in only 20.51%, suggesting that the hepatotoxicity was not more frequent than expected. Hepatotoxicity did not increase in proportion to the dose of leflunomide and MTX, age, gender, and disease activity. CONCLUSION: These results indicate that a combined treatment of leflunomide and MTX can be used safely by monitoring the liver enzyme, particularly in the first six months.

Serum methotrexate level is inversely related to the outcome of osteosarcoma patients / 소아과

PURPOSE: To evaluate the correlation between serum methotrexate (MTX) peak levels and clinical outcome of osteosarcoma, as well as to determine the correlation of these levels with the histologic response and event-free survival (EFS). METHODS: To maintain the homogeneity of the study population, we selected 52 patients with localized extremity osteosarcoma who had received two cycles of neoadjuvant chemotherapy consisting of high-dose (HD) MTX (12 g/m2), cisplatin (100 mg/m2), and doxorubicin (60 mg/m2). RESULTS: Totally, 204 courses of HD MTX were administered. The serial MTX levels (mean+/-SE) at 4 h (peak), 24 h, 48 h, and 72 h were 1292.14+/-12.83 micrometer, 9.29+/-3.89 micrometer, 1.73+/-1.37 micrometer, and 0.58+/-0.44 micrometer, respectively. The peak MTX serum level was 1292.14+/-12.83 micrometer. Neither the continuous average MTX peak level nor the dichotomized MTX peak level was related to the histologic response. However, the patients with a high 24-h MTX level (3.4 micrometer) had a poor histologic response (P=0.044). An inverse relationship was observed between MTX levels and survival: the EFS was better in the patients with a mean MTX peak level of less than 1,400 micrometer (P=0.002) and mean 24-h MTX level of less than 3.4 micrometer (P=0.011). CONCLUSION: The inverse correlation between the MTX level and the outcome is an unexpected finding. Further study on the pharmacokinetics of MTX is required to substantiate our findings and elucidate the mechanism involved.

Serum methotrexate level is inversely related to the outcome of osteosarcoma patients / 소아과

PURPOSE: To evaluate the correlation between serum methotrexate (MTX) peak levels and clinical outcome of osteosarcoma, as well as to determine the correlation of these levels with the histologic response and event-free survival (EFS). METHODS: To maintain the homogeneity of the study population, we selected 52 patients with localized extremity osteosarcoma who had received two cycles of neoadjuvant chemotherapy consisting of high-dose (HD) MTX (12 g/m2), cisplatin (100 mg/m2), and doxorubicin (60 mg/m2). RESULTS: Totally, 204 courses of HD MTX were administered. The serial MTX levels (mean+/-SE) at 4 h (peak), 24 h, 48 h, and 72 h were 1292.14+/-12.83 micrometer, 9.29+/-3.89 micrometer, 1.73+/-1.37 micrometer, and 0.58+/-0.44 micrometer, respectively. The peak MTX serum level was 1292.14+/-12.83 micrometer. Neither the continuous average MTX peak level nor the dichotomized MTX peak level was related to the histologic response. However, the patients with a high 24-h MTX level (3.4 micrometer) had a poor histologic response (P=0.044). An inverse relationship was observed between MTX levels and survival: the EFS was better in the patients with a mean MTX peak level of less than 1,400 micrometer (P=0.002) and mean 24-h MTX level of less than 3.4 micrometer (P=0.011). CONCLUSION: The inverse correlation between the MTX level and the outcome is an unexpected finding. Further study on the pharmacokinetics of MTX is required to substantiate our findings and elucidate the mechanism involved.

The comparison of beta-hCG levels according to treatment modalities in tubal pregnancy / 대한주산의학회잡지

OBJECTIVE: The aims of this study were to evaluate the effect of the methotrexate (MTX) method by comparing the change of the serum beta-hCG level between the MTX method and salpingectomy for treating tubal pregnancy, furthermore by analyzing differences between the MTX success group and the failure group, and to provide helps in establishing criteria for choosing the MTX method. METHODS: Medical records of the 118 patients who were diagnosed with tubal pregnancy were reviewed retrospectively for the period of January 2006 to December 2007 at Kangnam St. Marys Hospital. RESULTS: Between the MTX success group and the failure group, no difference was observed in pregnancy duration, but statistically significant differences were observed in the size of hematoma and the quantity of hemoperitoneum at the site of tubal pregnancy in ultrasonography and in the serum beta-hCG level (p<0.05). Compared with salpingectomy, the MTX method showed the patterns of a low decrease rate of serum beta-hCG level after treatment and even its increase during the early period of treatment, but the serum beta-hCG level decreased rapidly from Day 7 after treatment and became equal to that of salpingectomy on Day 28 after treatment. CONCLUSION: The size of hematoma and the quantity of hemoperitoneum at the site of tubal pregnancy and the serum beta-hCG level before treatment are important factors for success in the MTX method. The MTX method may be safe and effective relatively in hemodynamically stable tubal pregnancy patients, who need preserve the salpinx particularly.

Optimización de cultivos de hepatocitos humanos para estudios de citotoxicidad / Optimization of human hepatocyte cultures for citotoxicity studies

Los cultivos de hepatocitos entregan un valioso acercamiento al estudio de las funciones metabólicas específicas del hígado, evaluación de citotoxicidad. No existen líneas humanas inmortales con función normal. La inmortalización de hepatocitos humanos con el método UCHT1(medio de cultivo condicionado por células tumorales de tiroides) permitirá prolongar la sobrevida y función de estos, siendo útil para evaluar funcionalidad y citotoxicidad. Objetivo: Optimizar el cultivo de hepatocitos humanos. Metodología: En cultivos primarios de hepatocitos humanos, se agregó medio UCHT1 cultivando en superficies de colágeno, polilisina, gelatina y matrigel. Como control positivo, se utilizó línea Gherschenson (GER) para evaluar curva de crecimiento y producción de Glucógeno (PAS). Se evaluó citotoxicidad (LIVE/DEAD) en hepatocitos GER expuestos a Metotrexato (10, 100 y 1000 mM) a 24, 48 y 72 hrs. Resultados: Se realizó 3 cultivos primarios. Fue efectiva la utilización de Polilisina y Colágeno. Duración 8 meses. No se ha realizado la curva de crecimiento, ni evaluación de funcionalidad en hepatocitos humanos. La línea GER tiene un crecimiento exponencial (tiempo duplicación: 36 hrs). Se observó producción de glucógeno en condiciones de diferenciación hasta 120 hrs. La citotoxicidad por Metotrexato tiene una curva dosis dependiente, significativa en todas las concentraciones (p<0,001) (CL50 a 1000 mM a 24 hrs). Conclusiones: Se logró establecer una línea primaria de hepatocitos humanos. La polilisina y el colágeno han optimizado el establecimiento de cultivos primarios. El método PAS permitió evaluar producción de glucógeno (diferenciación). Los valores de citotoxicidad demostraron un efecto dosis dependiente en las condiciones experimentales. Logrando estandarizar el método para evaluación futura de líneas celulares humanas.

Background: Hepatocyte cultures are a valuable tool to study specific metabolic liver functions and cytoxicity. Human hepatocyte cell lines with normal function do not exist. Immortalization of human hepatocytes with a rat thyroid cell line (UCHT1) allows long-term survival and function of these cells, becoming useful to evaluate functionality and cytotoxicity. Aim: To optimize long-term culture of human hepatocytes. Material and Methods: UCHT1 media was added to primary cultures of human hepatocytes, seeding in collagen, gelatin, matrigel and polilisine surfaces. Gherschenson cell line (GER) was used as a positive control to evaluate the growth curve and Glycogen production (PAS). Cytotoxicity was evaluated (LIVE/ DEAD) in GER hepatocytes exposed to Metotrexate (10, 100 and 1000 µM) 24, 48 and 72 hrs. Results: Three primary cultures were made. The use of Polilisine and Collagen was effective. Cultures were kept for 8 months. Growth curves or evaluation of functionality in human hepatocytes, were not carried out. GER cell line had an exponential growth (duplication time: 36 hrs). Production of glycogen in differentiation conditions was observed up to 120 hrs. Cytotoxicity by Metotrexate had a dose dependent curve with a 50% lethal dose calculated as 1000 µM at 24 hrs. Conclusions: A primary line of human hepatocytes was obtained. Polilisine and collagen optimized the establishment of primary cultures. PAS method allowed the evaluation of glycogen production (differentiation). Cytotoxicity demonstrated a dose dependent effect in experimental conditions.

Detection of RFC,DHFR and GST-? expression in human osteosarcoma cell line with MTX-resistence by FQ-PCR / 中国矫形外科杂志

[Objective]To study the difference in expression of RFC, GST-?, DHFRmRNA between human osteosarcoma U2-OS cell line and the MTX-resisitant variants U2-OS/R1-R3, and to investigate the significance in MTX resistance for human osteosarcoma. [Methods]Three resistant MTX human osteosarcoma cell lines were established by pulse exposure parental cell line(U2-OS) in gradually increased dose of MTX . The expression of RFC,GST-?,DHFRmRNA were assayed by real-time fluorescence quantitative polymerase chain reaction(FQ-PCR).[Results]Three MTX-resistant variants(U2-OS/R1-R3) were successfully established , the results of the FQ-PCR revealed that the MTX resistance was associated with the decreased expression of the RFC mRNA and increased expression of DHFR mRNA and GST-? mRNA.[Conclusion]The author investigated the MTX resistant mechanism of human osteosarcoma cell line at a gene level. The decreased expression of RFC mRNA and the increased expression of DHFR mRNA and GST-? mRNA participate in the MTX resistance in human osteosarcoma cell lines U-2 OS. This provides the evidence for exploring the MTX resistance mechanism in clinical osteosarcoma patients ,and helps to screen the patients who are insensitive to MTX chemotherapy.

Research on combinative effect of methotrexate and caffeine on osteosarcoma cell line / 中国矫形外科杂志

[Objective]To observe whether or not caffeine can improve the cyto-toxicity effect of methotrexate(MTX) on osteosarcoma cell line.[Method]Osteosarcoma cell(OS-732)were incubated with no drug,caffeine,MTX or MTX adding caffeine separately,and were named as control group,caffeine group,MTX group and mixing group respectively.The ratios of cells on G2M phase of each group were measured with flow cytometry after 48 h incubation and the cell inhibition ratios were measured with the MTT colorimetric analysis after 72 h incubation.[Result]The ratios(%)of cells on G2M phase were control group(23.210?0.416),caffeine group(23.120?0.440),MTX group(28.770?0.531)and mixing group(0.105?0.002)(P