Determination of 5-Fluorouracil in Human Plasma by High-Performance Liquid Chromatography (HPLC)

5-Fluorouracil (5-FU) has a broad spectrum of anti-tumor activity, widely applied to the treatment of cancers. However, it is necessary to determine the plasma concentration of 5-FU in clinical practice due to its narrow therapeutic index. Therefore, a simple, economic and sensitive high-performance liquid chromatography (HPLC) method was developed and validated for the determination of 5-FU in human plasma. Ethyl acetate was chosen as extraction reagent. Chromatographic separation was performed on a Diamonsil C18 column (250 mm × 4.6 mm i.d., 5 μm) with the mobile phase consisting of methanol and 20 mmol/L ammonium formate using a linear gradient elution at a flow rate of 0.8 mL/min. 5-FU and 5-bromouracil (5-BU) were detected by UV detector at 265 nm. The calibration curve was linear over the concentration range of 5—500 ng/mL and the correlation coefficient was not less than 0.992 6 for all calibration curves. The intra- and inter-day precisions were less than 10.5% and 4.3%, respectively, and the accuracy was within ±3.7%. The recovery at all concentration levels was 80.1±8.6%. 5-FU was stable under possible conditions of storing and handling. This method is proved applicable to therapeutic drug monitoring and pharmacokinetic studies of 5-FU in human.

Embryoprotein TWIST distribution in breast cancer cells MDA-MB-231 treated with 5-Fluorouracil and malnutrition

TWIST is a transcription factor that belongs to the family of helix-loop-helix proteins involved in metastasis with essential role regulating cell movement during early development, as well as in the tumor progression and metastasis of many cancers including breast cancer. It will be interesting to study the relation among cancer chemotherapy, malnutrition and the transcription factors like TWIST in order to explore the risk to metastasis. We used breast cancer line MDA-MB-231. Cell cultures were treated with 5-Fluorouracil (5-Fu), as well as changes in serum and nonessentials amino acid (NEAA), to explore the cell viability and the cellular distribution of TWIST by immunocytochemistry. Our results indicate that cell viability decreased significantly with 5-Fu treatment whereas no changes were observed in malnutrition treatment. On the other hand, TWIST protein significantly increased its distribution in cytoplasm of treated groups with malnutrition as well as in those treated with 5-Fu compared with the control. These results suggest that TWIST translocation was modified by the treatments and further studies are necessary to suggest that TWIST could be a tag protein to avoid metastasis.

Davanat／5-FUⅡ期临床启动

美国Pro Pharmaceuticals公司称，Davanat／5-fluorouracil化疗药物的Ⅱ期临床试验已开始给第一位结直肠癌患者使用。该药物将在接受标准化疗方案后疾病仍有进展的结直肠癌转移患者中评估作为Ⅲ或Ⅳ线治疗方案的效果。

手术联合5-氟尿嘧啶与糖皮质激素治疗儿童腹股沟瘢痕疙瘩

目的探讨手术联合5-氟尿嘧啶(5-Fluorouracil,5-FU)与糖皮质激素注射治疗儿童腹股沟瘢痕疙瘩的效果。方法18例患者,共19个腹股沟瘢痕疙瘩,平均病史2.7年。手术切除瘢痕疙瘩,术后2～3周将0.05ml5-FU注射液与1ml复方倍他米松注射液和1ml2%利多卡因混合后适量注射入伤口内,以后逐渐降低5-FU和复方倍他米松注射液浓度并延长注射间歇期。平均治疗时间为8个月。结果手术联合5-FU与糖皮质激素混合注射治疗儿童腹股沟瘢痕疙瘩有效率为100%,其中完全治愈者72.2%,效果良好者27.8%。结论手术联合5-FU与糖皮质激素注射是治疗儿童腹股沟瘢痕疙瘩安全有效的方法。

Melatonin influences the biological characteristics of keloid fibroblasts through the Erk and Smad signalling pathways

Background:Keloids are abnormal fibrous hyperplasias that are difficult to treat.Melatonin can be used to inhibit the development of certain fibrotic diseases but has never been used to treat keloids.We aimed to discover the effects and mechanisms of melatonin in keloid fibroblasts(KFs).Methods:Flow cytometry,CCK-8 assays,western blotting,wound-healing assays,transwell assays,collagen gel contraction assays and immunofluorescence assays were applied to demonstrate the effects and mechanisms of melatonin in fibroblasts derived from normal skin,hypertrophic scars and keloids.The therapeutic potential of the combination of melatonin and 5-fluorouracil(5-FU)was investigated in KFs.Results:Melatonin significantly promoted cell apoptosis and inhibited cell proliferation,migration and invasion,contractile capability and collagen production in KFs.Further mechanistic studies demonstrated that melatonin could inhibit the cAMP/PKA/Erk and Smad pathways through the membrane receptor MT2 to alter the biological characteristics of KFs.Moreover,the combination of melatonin and 5-FU remarkably promoted cell apoptosis and inhibited cell migration and invasion,contractile capability and collagen production in KFs.Furthermore,5-FU suppressed the phosphorylation of Akt,mTOR,Smad3 and Erk,and melatonin in combination with 5-FU markedly suppressed the activation of the Akt,Erk and Smad pathways.Conclusions:Collectively,melatonin may inhibit the Erk and Smad pathways through the mem-brane receptor MT2 to alter the cell functions of KFs,while combination with 5-FU could exert even more inhibitory effects in KFs through simultaneous suppression of multiple signalling pathways.

Curcumin Reverses 5-Fluorouracil Resistance by Promoting Human Colon Cancer HCT-8/5-FU Cell Apoptosis and Down-regulating Heat Shock Protein 27 and P-Glycoprotein

Objective: To investigate the potential mechanisms that curcumin reverses 5-fluorouracil(5-FU) multidrug resistance(MDR). Methods: Cell growth and the inhibitory rate of curcumin(2–25 μg/mL) and/or5-FU(0.05–1000 μg/mL) on human colon cancer HCT-8 and HCT-8/5-FU(5-FU-resistant cel line) were determined using cel counting kit-8(CCK-8) assay. Apoptosis and cel cycle after 5-FU and/or curcumin treatment were detected by ?ow cytometry(FCM) and transmission electron microscopy(TEM). The expression of the multidrug resistance related factors p-glycoprotein(P-gp) and heat shock protein 27(HSP-27) genes and proteins were analyzed by reverse transcription polymerase chain reaction(RT-PCR) and Western blotting(WB), respectively. Results: The inhibitory rate of curcumin or 5-FU on HCT-8 and HCT-8/5-FU cells proliferation at exponential phase were in a dosedependent manner, HCT-8 cell line was more sensitive to curcumin or 5-FU when compared the inhibitory rate of HCT-8/5-FU. The 50% inhibitory concentration(IC50) of combination 5-FU and curcumin(4.0 μg/mL)in HCT-8/5-FU was calculated as 179.26 μg/mL, with reversal fold of 1.85. Another IC50 of combination 5-FU and curcumin(5.5 μg/mL) in HCT-8/5-FU was calculated as 89.25 μg/mL, with reversal fold of 3.71. Synergistic effect of 5-FU and curcumin on HCT-8 and HCT-8/5-FU cells were found. The cell cycle analysis performed by FCM showed that HCT-8 and HCT-8/5-FU cel s mostly accumulated at G0/G1 phase, which suggested a synergistic effect of curcumin and 5-FU to induce apoptosis. FCM analysis found that the percentage of apoptosis of cel s treated with curcumin, 5-FU and their combination were signi?cantly increased compared to the control group(P<0.05), and the percentage of apoptosis of the combination groups were slightly higher than other groups(P<0.05). The m RNA levels of P-gp(0.28±0.02) and HSP-27(0.28±0.09) in HCT-8/5-FU cel s treated with combination drugs were lower than cel s treated with 5-FU alone(P-gp, 0.48±0.07, P=0.009;HSP-27, 0.57±0.10, P=0.007). The protein levels of P-gp(0.25±0.06) and HSP-27(0.09±0.02) in HCT-8/5-FU cells treated with combination drugs were decreased when compared to 5-FU alone(P-gp, 0.46±0.02, P=0.005;HSP-27, 0.43±0.01, P=0.000). Conclusions: Curcumin can inhibit the proliferation of human colon cancer cells. Curcumin has the ability of reversal effects on the multidrug resistance of human colon cancer cells lines HCT-8/5-FU. Down-regulation of P-gp and HSP-27 may be the mechanism of curcumin reversing the drug resistance of HCT-8/5-FU to 5-FU.

Near-infrared responsive 5-fluorouracil and indocyanine green loaded MPEG-PCL nanoparticle integrated with dissolvable microneedle for skin cancer therapy

The prevalence of skin cancer is rising along with the rapid population aging in recent years.Traditional therapies,such as surgical treatment,radiotherapy,chemotherapy,photodynamic therapy,and immunotherapy,may accompany serious side effects,limiting their clinical benefits.According to the biological characteristics of skin cancer,we have already established two kinds of synergetic systems of photothermal therapy(microneedle)and chemotherapy,containing gold nanorods(GNR).Although the microneedle system exhibited great potential for skin cancer treatment,the system could be still improved further.So,we designed a near-infrared lightresponsive 5-fluorouracil(5-Fu)and indocyanine green(ICG)loaded monomethoxy-poly(ethylene glycol)-polycaprolactone(MPEG-PCL)nanoparticle(5-Fu-ICG-MPEG-PCL),and then 5-Fu-ICG-MPEG-PCL was integrated with a hyaluronic acid dissolvable microneedle system(HA MN)to get 5-Fu-ICG-MPEG-PCL loaded HA MN for treating skin cancers,including human epidermoid cancer and melanoma.In this system,hyaluronic acid,the microneedle carrier,possesses good skin penetration ability and is approved by FDA as a pharmaceutical adjuvant;5-Fu is recommended by FDA for skin cancer treatment;ICG,a photothermal agent,possesses a strong photothermal ability and is approved by FDA for its use in the human body.We hypothesized that 5-Fu-ICG-MPEG-PCL could be delivered by the dissolvable microneedle through the skin,and the release behavior of the drug in the nanoparticle could be controlled by near-infrared light for achieving a single-dose cure of skin cancer,improving the cure rate of skin cancer and providing a new idea and possibility for the clinical treatment of skin cancer.

低浓度5-氟尿嘧啶与糖皮质激素联合激光治疗瘢痕疙瘩的疗效

目的 观察低浓度5-氟尿嘧啶(5-Fu)与糖皮质激素局部注射联合激光治疗瘢痕疙瘩的疗效及安全性.方法 瘢痕疙瘩70例,102处病灶,平均病史8.6年,平均治疗时间为10.29个月.70例瘢痕疙瘩患者108处病灶进行单纯瘢痕内注射,0.6 ml 5-Fu与5 ml曲安奈德、1 ml 2%利多卡因混合后局部注射于瘢痕全层注射,每2～4周1次,瘢痕完全萎缩后逐渐降低药物浓度并延长注射间歇期.另一组70例瘢痕疙瘩患者102处病灶将三联药物局部注射半个月后Nd:YAG激光照射,每2～4周1次.结果 在治疗6个月以上的患者中,5-氟尿嘧啶与糖皮质激素联合激光治疗瘢痕疙瘩的总有效率为97.06%,其中完全缓解者占45.10%,极大缓解者占49.02%,部分缓解者占2.94%,未缓解者为2.94%.结论 低浓度5-氟尿嘧啶与糖皮质激素局部注射联合激光治疗瘢痕疙瘩的疗效显著.