秦川牛宰后成熟过程中线粒体Tu翻译延长因子对牛肉持水性的影响

探究秦川牛宰后成熟过程中线粒体Tu翻译延长因子(mitochondrial Tu translation elongation factor,TUFM)表达对肉的持水性影响。以秦川牛背最长肌为研究对象,测定4℃不同成熟时间下的pH值、贮藏损失、离心损失、蒸煮损失、水分分布、肌原纤维蛋白等指标变化情况,测定不同成熟时间(0、96、192 h)下TUFM表达量及其含量、Beclin1蛋白表达量。结果显示:在秦川牛宰后成熟期间,肌原纤维蛋白发生降解,TUFM的表达量与Beclin1蛋白表达量和牛肉的持水性存在密切关系,其中蛋白质组学测定的TUFM表达量变化与TUFM含量变化趋势一致,Beclin1蛋白表达量、贮藏损失、离心损失、蒸煮损失整体均呈先上升后下降趋势,pH值呈先下降后上升趋势;Pearson相关性分析表明,牛背最长肌中TUFM表达量与低场核磁共振峰面积比P_(2b)、Beclin1蛋白表达量呈极显著正相关(P<0.01),与贮藏损失、离心损失、蒸煮损失呈显著正相关(P<0.05),与P_(21)呈极显著负相关(P<0.01),与P_(22)呈显著负相关(P<0.05),与pH值无显著相关性(P>0.05)。通过蛋白质组学鉴定出23种与TUFM相关的差异蛋白,通过基因本体论、京都基因与基因组百科全书通路分析发现,差异蛋白可通过多种途径参与能量代谢,进而介导细胞自噬;对差异蛋白和持水性指标进行Pearson相关性分析发现,有5种差异蛋白(ATP5F1D、EEF1A2、GSPT1、NDUFB5、SUCLG1)与持水性指标具有显著相关性(P<0.05、P<0.01)。分析可知,包括TUFM在内,共6种蛋白主要通过能量代谢和氧转运等途径正向或负向影响细胞自噬,从而影响肉的持水性。

Conservative management of cervical pregnancy with intramuscular administration of methotrexate and KCl injection: Case report and review of the literature

We report the case of a cervical pregnancy successfully treated with intramuscular injection of methotrexate(MTX) and intramniotic administration of potassium chloride. A 41-year-old woman was admitted to our Department with the suspicion of ectopic pregnancy. Transvaginal ultrasound revealed empty endometrialcavity, gestational sac within the cervical canal and embryonic echo measuring crown rump length 1.5 mm. Serum beta human chorionic gonadotropine(β-HCG) was measured 28590 IU/L. No cardiac activity was detected. The diagnosis of a cervical pregnancy was made. Patient was treated with intramuscular administration of methotrexate(50 mg/m2) in combination with ultrasoundguided intramniotic injection of KCl(2 meq/mL). Gradual decrease of β-HCG levels as well as ultrasound observation of collapsed gestational sac was observed. No curettage was necessitated. Patient was discharged on day 10 th and was set in follow-up on a weekly basis. β-HCG values were measured < 10 IU/L on 56 th day after MTX administration. Intramuscular administration of MTX may be effective in treatment of cervical pregnancy without additional interventional measures.

High-dose methotrexate and zanubrutinib combination therapy for primary central nervous system lymphoma

In this editorial I comment on the article,published in the current issue of the World Journal of Clinical Oncology.Primary central nervous system lymphoma(PCNSL)is a disease of elderly and immunocompromised patients.The authors reported clinical results of 19 patients with PCNSL treated with zanubrutinib/high dose methotrexate(HD-MTX)until disease progression.They demonstrated that the combination of zanubrutinib with HD-MTX led to a marked clinical response and tolerability among these patients.They also observed that cerebrospinal fluid liquid biopsy to detect circulating tumor DNA may be a good option for evaluating treatment response and tumor burden in patients with PCNSL.PCNSL is a challenging disease for treatment as these patients present with different neurological states and comorbidities.Treatment has evolved over the years from whole brain radiotherapy to HD-MTX followed by autologous stem cell transplant.Gradually,treatment of patients with PCNSL is going to become individualized.

Intratubal Methotrexate Injection Combined with Chinese Herbal Medicine for Tubal Pregnancy and Following Pregnancy Prognosis

Objective: To compare the effects of treatment of tubal pregnancy (TP)and its following second pregnancy by intratubal methotrexate injection(IMI) alone and combination of IMI with Chinese herbal medicine. Methods: Thirty-five patients suffering from unruptured TP were divided into two groups at random, to the 19 patients in the treated group, the treatment of combined IMI with Ectopic Pregnancy decoction No. 2 (EP2, a traditional Chinese medical decoction) was applied, and to the other 16 patients in the control group, IMI alone was applied for control. Serum concentrations of human chorionic gonadotro-pinβ(β-HCG), size of the gestational sac, existent time of fetal cardiac beat and peritoneal fluid were measured before and after treatment. And hysterosalpingography were performed 6 months after ending the treatment to verify the presence of tubal obstruction and the condition of relapse.Results: The treatment of all the 35 women was successful. The recovery duration of serumβ-HCG, disappearance duration of TP sac and existent time of peritoneal fluid in the treated group were 20.0±7. 8 days, 1.2±0. 7 months and 10. 7±2. 9 days respectively, which were significantly different from those in the control group (24. 4 ±8.1 days, 3.6±1.7 months and 19.1±3. 2 days respectively(P<0. 05, P<0.01 and P<0. 05 respectively), but the existent time of fetal cardiac beat in the two groups (8.8±1. 9 days vs 9.0±1. 3 days) was not significantly different (P>0.05). The post-treatment oviduct obstructive rate in the two groups was 10.5% and 43.8% respectively, that in the treatment group was less significant (P<0.05). The relapse rate of EP in the treatment group was insignificantly different from that in the control group (5.3% vs 18.8%, P>0.05). Conclusion: The two therapies (IMI alone and IMI combined with EP2) could obtain e-qual efficacy in curing TP. Compared with IMI alone, the combined therapy appears to have the effects of accelerating the resorption of gestational sac and peritoneal fluid, improving the patency of fallopian tube and ameliorating the circumstance of pregnancy, which is favorable to improvement of the re-pregnancy rate and reduction of the re-occurrence of ectopic pregnancy as well as to the enhancement of the effect of IMI in killing trophocytes. But there is not enough proof to show the potency of EP2 in killing embryo.

Perioperative Use of Methotrexate and Tumor Necrosis Factor α Inhibitors Combination Therapy Is Not Likely to Increase Post-Operative Infection Rate in the National Veterans Health Administration Administrative Databases

Objective: The aim of the study is to assess the risk of post-operative outcome in rheumatoid arthritis (RA) patients continuing versus stopping combination therapy of methotrexate (MTX) and hydroxychloroquine (HCQ) or tumor necrosis factor α inhibitors (TNF) prior to surgery. Methods: Using the United States Veterans Affairs (VA) databases, we identified surgical procedures in a 17-year cohort of RA patients. Among those patients, those on MTX + HCQ or MTX + TNF were identified. Post-operative outcome variables include infection, length of post-operative hospital stay and death. Results: We identified a total of 29,708 surgeries in RA patients. Among them, we identified the most recent elective surgeries without pre-operative infection in 16,174 patients. There were 783 and 550 patients on MTX + HCQ and MTX + TNF, respectively. The rates of post-op infection were 5% and 4% for the MTX + HCQ and MTX + TNF continuing medication groups, respectively, similar to the general RA population (5%). Sensitivity analyses at various time points of discontinuation combination therapies prior to surgery did not show significant change in terms of infection. Conclusions: The prevalence of adverse outcome is low. The proportion of post-operative infection in continuing and discontinuing medicine groups is similar for both MTX + HCQ and MTX + TNF. While we were unable to formally compare proportions of post-operative infection among the two groups, these preliminary findings do not support the hypothesis that continuing either MTX + TNF or MTX + HCQ combination during perioperative period increases post-operative infection compared with discontinuation prior to therapy.

Status of Renal and Liver Function in Rheumatoid Arthritis (RA) Patients of Chattogram, Bangladesh Treated with Methotrexate (MTX)

Rheumatoid Arthritis (RA) is a chronic autoimmune disorder that is usually manifested as inflammation in multiple joints and several extra-articular symptoms, involving the liver, kidney, eye, skin, blood, blood vessels, heart, lungs, nervous system, and other organs. Methotrexate (MTX) is the anchor drug that treats RA. As renal and liver abnormalities are more common during disease conditions as well as during the treatment period, we tried to find out if there is any impact of MTX in these organs during the treatment of RA patients. Once the disease complications are developed, it is quite difficult to reverse the disease, and treatment in this situation is not very effective. Consequently, patients suffer a lot. So, early evaluation of renal and liver function is essential for the treatment of RA patients and it might also help prevent different complications which are usually very frequently observed. This was a cross-sectional study. A total of 150 RA patients treated with MTX were evaluated for the study where female and male respondents were 115 and 35 respectively. In this study, we found that 82% of RA patients had creatinine levels ≤ 1.1 mg/dL although the normal range of serum creatinine is below 1.4 mg/dL. Usually, a 15% increase in Serum creatinine level from the baseline is considered renal impairment. We found 4% of such cases. Moreover, 2% of RA patients had creatinine levels above the normal range of 1.4 mg/dL and those patients were hypertensive as well. So, a total (4 + 2 = 6)% had renal impairments. Among them, 5% had diabetes mellitus. On the other hand, the ultrasonogram (USG) of RA patients with kidney disease showed signs of renal parenchymal disease and 3% of RA patients having renal problems whose serum creatinine level was within the normal range showed signs of chronic kidney disease (CKD). On the other hand, 2% of RA patients showed signs of hepatic parenchymal disease. In this study, 69% of RA patients had ALT levels ≤ 50 mg/dL, 23% had 50 - 100 mg/dL, and 5% had 101 - 150 mg/dL. The remaining 3% of RA patients had ALT levels above 150 mg/dL. All those patients with ALT levels above 100 mg/dL used Nonsteroidal anti-inflammatory drugs (NSAIDs) concomitantly. Different parameters of liver and renal function should be monitored strongly in RA patients treated with MTX and NSAIDs. MTX should not be given for a prolonged period without monitoring renal and liver function. As MTX, Diabetes Mellitus, Hypertension, etc., may cause renal complications, we could not concretely conclude which one is the actual causative agent.

Successful treatment of a high-risk nonseminomatous germ cell tumor using etoposide, methotrexate, actinomycin D, cyclophosphamide, and vincristine: A case report

BACKGROUND Choriocarcinoma is an infrequent entity and the most aggressive subtype of germcell tumors.Because of early metastatic spread and rapid disease progression,choriocarcinoma patients display poor prognosis.Although etoposide,methotrexate,actinomycin D,cyclophosphamide,and vincristine(EMA-CO)regimen is widely used to treat gestational trophoblastic tumors in females,its role in treating male choriocarcinoma is seldom reported.CASE SUMMARY A 32-year-old man was diagnosed with burned-out primary germ cell tumors(GCT)with retroperitoneum,liver and lung metastases.Biopsy of the liver revealed pure choriocarcinoma.The patient received bleomycin,etoposide,and cisplatin chemotherapy.After two cycles of treatment,response evaluation revealed the mixed response.EMA-CO regimen was used in the second-line therapy.After eight cycles,the patient showed a potentially resectable state and thus,all residual masses were surgically removed.The patient was completely cured,and 10 years later,he is leading a healthy life without complications.CONCLUSION This paper is the first case of high-risk nonseminomatous GCT in a male patient to be successfully treated with the EMA-CO regimen.The EMA-CO regimen can be used actively in patients with high-risk nonseminomatous GCT.

Study on the Regulation of Qinxitong Combined with Methotrexate Tablets on Multiple Inflammatory Indicators and Mechanism Factors in Rheumatoid Arthritis

Objective:To study the changes of several inflammatory index mechanism factors in the clinical efficacy of tablet and methotrexate in rheumatoid arthritis.Methods:20 patients with rheumatoid arthritis were randomly divided into observation(n=10)and control(n=10),then normal people as normal(n=10),all three groups were given methotrexate and the observation group were treated with Qin interest pain tablets.Expression of inflammatory index mechanism factors in each group.Results:After treatment,inflammatory index mechanism factors were detected;a significant decrease of IFN-g,IL-1b,IL-10,IL-37,TNF-a and other factors was found before and after combination treatment(P<0);IL-8 and IL-13 expression(P<0);the CRP,ESR score of the study group was lower than the control group(P<0.05);and the inflammatory index mechanism factors were affected by drug combination.Conclusion:Tablets combined with methotrexate in rheumatoid arthritis patients is better than methotrexate.It obviously changed the expression of the patient’s inflammatory index mechanistic factors,which has a regulatory effect on the inflammatory mechanistic factors.

Bovine Serum Albumin (BSA) Microspheres Containing Methotrexate (MTX) I.Pharmaceutical Aspects

Microspheres Ⅰ,Ⅱ and Ⅲ were produced by emulsion technique.Microsphere I was solidified by glutaraldehyde crosslinking,microsphere Ⅱ was solidified by glutaraldehyde crosslinking and further treated with glycine solution and microsphere Illwas solidified by heating denaturation only.The results showed that the microsphere diameter produced by cross[inking was bigger than that prepared by heating.The microsphere Ⅱ had higher hydrophilicity than Microsphere I had.The methotrexate (MTX) contents in microspheres Ⅰ and Ⅱ were 2.73±0.053%,2.87±0.119% respectively. microsphere Ⅲ was only blank microspheres with MTX adsorbed on their surfaces.In vitro release studies,microspheres I and I have maintained sustained release of MTX till the next day,it was found that the drug releases from microspheres Ⅰ and Ⅱ were governed by Higuchi diffusion law.

工程教育中心何以推动科教融合——荷兰4TU工程教育中心的探索性单案例研究

工程教育中心作为建立在大学或研究机构中的跨学科交叉合作平台,是连接科学研究与教育实践的纽带,在高质量工程人才培养中发挥着重要作用。荷兰4TU工程教育中心利用4所顶尖理工大学在工程学科和教育领域的独特优势,积极与研发单位、教育单位、企业部门合作,通过将前沿科学研究彻底融入工程课程设计、教学模式等多个方面,形成了独具一格的科教融合工程人才培养模式。文章从战略目标、组织架构、运行机制、质量保障4个维度详实分析了4TU工程教育中心推动科教融合的内在机制,总结归纳其在主题项目设置、教育共同体形成、课程体系迭代、创新网络构建、内外部质量保障等方面的核心特征,期望对我国科教融合的工程教育改革与建设有所启示。

Analytical methodologies for determination of methotrexate and its metabolites in pharmaceutical,biological and environmental samples

Methotrexate(MTX)is a folate antagonist drug used for several diseases,such as cancers,various malignancies,rheumatoid arthritis(RA)and inflammatory bowel disease.Due to its structural features,including the presence of two carboxylic acid groups and its low native fluorescence,there are some challenges to develop analytical methods for its determination.MTX is metabolized to 7-hydroxymethotrexate(7-OH-MTX),2,4-diamino-N10-methylpteroic acid(DAMPA),and the active MTX polyglutamates(MTXPGs)in the liver,intestine,and red blood cells(RBCs),respectively.Additionally,the drug has a narrow therapeutic range;hence,its therapeutic drug monitoring(TDM)is necessary to regulate the pharmacokinetics of the drug and to decrease the risk of toxicity.Due to environmental toxicity of MTX;its sensitive,fast and low cost determination in workplace environments is of great interest.A large number of methodologies including high performance liquid chromatography equipped with UVevisible,fluorescence,or electrochemical detection,liquid chromatography-mass spectroscopy,capillary electrophoresis,UVevisible spectrophotometry,and electrochemical methods have been developed for the quantitation of MTX and its metabolites in pharmaceutical,biological,and environmental samples.This paper will attempt to review several published methodologies and the instrumental conditions,which have been applied to measure MTX and its metabolites within the last decade.

Risk of liver disease in methotrexate treated patients

Methotrexate is the first line drug treatment for anumber of rheumatic and non-rheumatic diseases. It is effective in controlling disease activity and preventing disease-related damage, and significantly cheaper than many alternatives. Use in rheumatoid arthritis infers a significant morbidity and mortality benefit. Methotrexate is generally well tolerated but can cause symptomatic adverse events. Multiple serious adverse events have been attributed to methotrexate, based largely on older reports using high or daily doses, and subsequent case reports and circumstantial evidence. The risk with modern dosing regimens: Lower doses, weekly schedules, and concomitant folic acid is less clear. Clarification and dissemination of the actual risk is crucial so appropriate judgements can be made for patients who may benefit from this treatment. Methotrexate has been associated with a range of liver related adverse events ranging from asymptomatic transaminase elevations to fibrosis and fatal hepatic necrosis. Concern over potential liver toxicity has resulted in treatment avoidance, cessation, or recommendations for investigations which may be costly, invasive and unwarranted. Modern laboratory monitoring of liver blood tests may also influence the risk of more serious complications. The majority of present day studies report an approximate doubling of the relative risk of elevated transaminases in methotrexate treated patients but no increased risk of symptomatic or severe liver related adverse events. In this article we will review the evidence around methotrexate and liver related adverse events.

Use of methotrexate in inflammatory bowel disease in 2014: A User's Guide

Methotrexate has been used an immunomodulator in many autoimmune diseases,including inflammatory bowel disease. However,many physicians are unfamiliar or uncomfortable with its use in the management of inflammatory bowel disease. We summarize the data for use of methotrexate in common clinical scenarios:(1) steroid dependant Crohn's disease(CD);(2) maintenance of remission in steroid free CD;(3) azathioprine failures in CD;(4) in combination therapy with Anti-TNF agents in CD;(5) decreasing antibody formation to Anti-TNF therapy in CD;(6) management of fistulizing disease in CD; and(7) as well as induction and maintenance of remission in ulcerative colitis. An easy to use algorithm is provided for the busy clinician to access and safely prescribe methotrexate for their inflammatory bowel disease patients.

Cytoprotective effects of amifostine,ascorbic acid and N-acetylcysteine against methotrexate-induced hepatotoxicity in rats

AIM: To investigate the potential role of oxidative stress and the possible therapeutic effects of N-acetyl cysteine (NAC), amifostine (AMF) and ascorbic acid (ASC) in methotrexate (MTX)-induced hepatotoxicity.

Protective effects of Balanites aegyptiaca extract, melatonin and ursodeoxycholic acid against hepatotoxicity induced by methotrexate in male rats

Objective: To compare the degree of ameliorative effects of Melatonin(MEL), Ursodeoxycholic acid(UDCA) and Balanites aegyptiaca(BA) against hepatotoxicity induced by MTX for one month. Methods: Eighty adult male rats(Sprague Dawely) weighing(190±10g), were randomly divided into eight equal groups: Control, MTX, MEL, BA, UDCA, MTX+MEL, MTX+BA, MTX+UDCA. Liver function biomarker enzymes, liver tissue oxidative stress parameters, together with total antioxidant capacity and tumor necrosis factor(TNF-α) were determined. Histopathological and immunohistochemistry examinations for TNF-α were also done. Results: MTX showed significant increase in alanine transaminase(ALT), aspartate transaminase(AST), alkaline phosphatase(ALP), gamma glutamyl transferase(GGT), total and direct bilirubin, as well as TNF-α levels, oxidized glutathione(GSSG), malodialdehyde(MDA) and nitric oxide(NO). whereas, total protein, albumin, total antioxidant capacity, reduced glutathione(GSH), glutathione peroxidase(GPx), glutathione reductase(GR), glutathione S-transferase(GST), superoxide dismutase(SOD) and catalase(CAT) levels were significantly decreased in MTX treated group. These alterations were improved by MEL and BA treatment, whereas no improvement was noticed in UDCA treatment. Conclusions: BA may be as promising as MEL in the hepatoprotection against MTX toxicity through their antioxidant and radical scavenging activities. In addition, it is not recommended to co-administer UDCA with MTX as it enhanced inflammation and damage to the liver.

Prevalence and risk factors of methotrexate hepatoxicity in Asian patients with psoriasis

AIM: To establish the prevalence of liver fibrosis and to evaluate the possible risk factors for fibrosis and progression in Asian with psoriasis treated with methotrexate (MTX) based on liver histology. METHODS: Patients with psoriasis treated with MTX referred to the Department of Gastroenterology, Tan Tock Seng Hospital for liver biopsy were identified and retrospectively studied. Patient case notes and electronic records were retrieved from the hospital database and relevant data collated. Histological changes of liver biopsies were staged according to Roengik score. The factors assessed were age, gender, ethnicity, cumulative dose of MTX, presence of comorbid conditions such as diabetes, hypertension, hyperlipidemia, and ethanol use. We also assessed the histological change in those with multiple liver biopsies. Statistical analysis was performed using Stata V.9.2. RESULTS: There were altogether 59 patients (median age 50 years old, range 22-81 years old, male, 88%) with 98 biopsies liver biopsies; 6 normal [median cumulative dose (MCD), 2285 mg]; 62 gradeⅠ (MCD 2885 mg), 23 grade Ⅱ (MCD 1800 mg) and 7 grade Ⅲ (MCD 1500 mg). There was no grade Ⅳ or cirrhosis. The prevalence of liver fibrosis (grade Ⅲ) was 12%. Of the factors assessed, diabetes (P=0.001) and hypertension (P=0.003) were significant for fibrosis on univariate analysis but not on multivariate analysis. Of the 26 patients who had more than one biopsy (median 2, range 2-6), 57.7% (n=15) were stable, 34.6% (n=9) had progression and 7.7% (n=2) had regression of histological grades. On univariate analysis, nonChinese ethnicity (P=0.031), diabetes (P=0.018), and hyperlipidemia (P=0.011) were predictive of progression of grades, but these were not significant on multivariate analysis. CONCLUSION: Liver fibrosis in Asian psoriatic population on MTX is comparable to the West. Cumulative dose was not associated with liver fibrosis. Metabolic syndrome is important factors.

Degradation of methotrexate by UV/peroxymonosulfate:Kinetics,effect of operational parameters and mechanism

Methotrexate(MTX)is one of the most consumed anti-cancer drugs in the pharmaceutical market around the world.The widespread occurrence of MTX in aquatic environment through hospital effluent has attracted increasing concern due to its potential to induce water pollution.In the present study,the degradation of MTX in aqueous medium was investigated by UV-activated peroxymonosulfate(PMS).A significant improvement in degradation rate by increasing UV intensity and PMS concentration while the decrease in degradation efficiency with the increase of solution p H and initial concentration of MTX was observed.The proposed UV/PMS process could achieve more than 90%MTX degradation in 30 min with a good mineralization degree(65%).A pseudofirst order kinetic model was employed and successfully predicted the degradation of MTX.The effect of other operational parameters such as the initial concentration of the targeted compound,dosage of oxidant(PMS),solution p H and UV intensity on the degradation rate were investigated.At the last,the main transform intermediates were identified using LC–MS and possible degradation pathways were proposed.The results show that UV/PMS can be used as an efficient technology to treat pharmaceuticals such as methotrexate containing water and wastewater.

Methotrexate combined with methylprednisolone for the recovery of motor function and differential gene expression in rats with spinal cord injury

Methylprednisolone is a commonly used drug for the treatment of spinal cord injury, but high doses of methylprednisolone can increase the incidence of infectious diseases. Methotrexate has anti-inflammatory activity and immunosuppressive effects, and can reduce in- flammation after spinal cord injury. To analyze gene expression changes and the molecular mechanism of methotrexate combined with methylprednisolone in the treatment of spinal cord injury, a rat model of spinal cord contusion was prepared using the PinPointTM preci- sion cortical impactor technique. Rats were injected with methylprednisolone 30 mg/kg 30 minutes after injury, and then subcutaneously injected with 0.3 mg/kg methotrexate 1 day after injury, once a day, for 2 weeks. TreadScan gait analysis found that at 4 and 8 weeks after injury, methotrexate combined with methylprednisolone significantly improved hind limb swing time, stride time, minimum longitudinal deviation, instant speed, footprint area and regularity index. Solexa high-throughput sequencing was used to analyze differential gene ex- pression. Compared with methylprednisolone alone, differential expression of 316 genes was detected in injured spinal cord treated with methotrexate and methylprednisolone. The 275 up-regulated genes were mainly related to nerve recovery, anti-oxidative, anti-inflammatory and anti-apoptotic functions, while 41 down-regulated genes were mainly related to proinflammatory and pro-apoptotic functions. These results indicate that methotrexate combined with methylprednisolone exhibited better effects on inhibiting the activity of inflammatory cytokines and enhancing antioxidant and anti-apoptotic effects and thereby produced stronger neuroprotective effects than methotrexate alone. The 316 differentially expressed genes play an important role in the above processes.

Prediction of Response of Collagen-induced Arthritis Rats to Methotrexate： An ~1H-NMR-based Urine Metabolomic Analysis

Over one half the patients with rheumatoid arthritis （RA） are being treated with methotrexate （MTX）. Although well proven, the efficacy of MTX varies in individual patients. This study examined the metabolic biomarkers that can be used to predict the therapeutic effect of MTX by using metabolomic analysis. Rats were immunized with collagen to rapidly cause collagen-induced arthritis （CIA） and then treated with 0.1 mg/kg MTX for 4 weeks. The clinical signs and the histopathological features of CIA were observed to evaluate the therapeutic effects. Urine samples of CIA rats were collected, and analyzed by using 600 M 1H-nuclear magnetic resonance （1H-NMR） for spectral binning after the therapy. The urine spectra were divided into spectral bins, and 20 endogenous metabolites were assigned by Chenomx Suite. Multivariate analyses were performed to identify the spectral pattern of endogenous metabolites related to MTX therapy. The results showed that the clustering of the spectra of the urine samples from the responsive rats （n=20） was different from that from the non-responsive rats （n=11）. Multivariate analysis showed difference in metabolic profiles between the responsive and non-responsive rats by using partial least squares-discrimination analysis （PLS-DA） （R2=0.812, Q2=0.604）. In targeted profiling, 13 endogenous metabolites （uric acid, taurine, histidine, methionine, glycine, etc.） were selected as putative biomarkers for predicting therapeutic response to MTX. It was suggested that 1H-NMR-based metabolomic analysis can be used to predict the therapeutic effect of MTX, and several metabolites were found to be related to the therapeutic effects of MTX.

Two Cases of Primary Intraocular Lymphoma:Fine Needle Aspiration Diagnosis and Intravitreal Methotrexate Treatment

We described clinical process of two cases of intraocular lymphoma in aspects of early diagnosis by fine needle aspiration（FNA） and biopsy and treatment by intravitreal methotrexate（MTX）.Two patients were suspected to have primary intraocular lymphoma（PIOL） with geographic yellow-white infiltrates and vitreous opacity.FNA confirmed malignant intraocular lymphoma in one patient and failed in the other patient due to complication of vitreous hemorrhage.Subsequent vitreous biopsy confirmed malignant intraocular lymphoma in the other patient.Both patients were treated by intravitreal methotrexate.In case 1 the tumor had complete remission and follow-up of 12 months had not found any signs of recurrence.In case 2 the patient died of brain metastasis 22 months after the ocular biopsy.Our findings demonstrate that although cytological examination of vitrectomy specimens remains the gold standard in diagnosis of PIOL,examination of FNA and biopsy increases the reliability of early diagnosing or excluding a PIOL.Individualized intravitreal methotrexate can be used to effectively treat PIOL.More effective integrated program treating primary central nervous system lymphoma/PIOL is worthy of looking forward to.