丝裂霉素C眼用凝胶的制备及质量控制

制备丝裂霉素C眼用凝胶剂并建立其质量控制方法.以丝裂霉素为主药,以卡波姆和羧甲基纤维素( HPMC K4M)为基质制备眼用凝胶,采用HPLC法测定制剂中丝裂霉素C的含量.所制得的眼用凝胶为淡紫色透明状,丝裂霉素C在21 - 210μg/ml范围内呈良好的线性关系,A=0.482C+ 0.009,精密度RSD为0.049...     

加替沙星眼用凝胶的制备及质量控制

介绍了加替沙星眼用凝胶的制备,以HPLC法测定制剂中加替沙星的含量及有关物质,平均回收率为99.46％,RSD％:0.53％,3批样品的含量分别为:99.98％,99.92％,99.90％,有关物质为:0.236％,0.246％,0.237％.该法适用于加替沙显眼用凝胶质量控制.     

清开灵眼用凝胶的制备与质量控制

研发清开灵眼用凝胶并对其质量进行控制。以羟丙甲纤维素（HPMC）主要基质刺成眼用凝胶,采用定性和定量方法进行质量控制,定量方法采用紫外可见分光光度仪测定其主要有效成分：黄芩苷和胆酸的含量。清开灵眼用凝胶成型性、澄明度好,且成本低,质量可控。本方法简便、准确、重现性好,可用于清开灵眼用凝胶的质量控制。     

血管抑素联合丝裂霉素C防止原发性翼状胬肉术后复发的临床疗效观察

目的　观察血管抑素（Angiostatin AS）联合丝裂霉素C（Mitomycin MMC）防止原发性翼状胬肉术后复发的临床疗效。 方法 对365例（369眼）原发性翼状胬肉随机分成3组,分别采用A组行逆行性翼状胬肉切除术联合丝裂霉素（121眼）,B组行逆行性翼状胬肉切除术联合血管抑素（122眼）,C组行逆行性翼状胬肉切除术联合丝裂霉素及血管抑素（126眼）等方法治疗,随访12～18个月,比较其复发率。 结果 3组间差异具有显著性意义（P<0.05）,C组复发率（0.8%, 1/126）最低,其次是B组（5.7%, 　7/122）,最后是A组（7.4%, 9/121）。 结论 逆行性翼状胬肉手术联合丝裂霉素C及血管抑制素治疗原发性翼状胬肉,其疗效显著,复发率低,值得推广。     

自体角膜缘干细胞移植联合丝裂霉素C治疗翼状胬肉疗效观察

目的：观察自体角膜缘干细胞移植术联合丝裂霉素C应用治疗翼状胬肉的疗效。方法对102例122只眼行胬肉切除,将同侧眼颞上方带有角膜缘干细胞的结膜瓣移植到胬肉处的巩膜上,术中联合应用0.2mg/ml丝裂霉素C治疗。结果术后反应轻,移植片存活,4只眼复发,复发率为3.3%。结论自体角膜缘干细胞移植术联合丝裂霉素C应用治疗翼状胬肉,方法简单,恢复角膜缘屏障,降低了复发率,是安全、有效的方法。     

角膜缘干细胞移植联合丝裂霉素C治疗复发性翼状胬肉

目的 观察自体角膜缘干细胞移植联合丝裂霉素C治疗复发件翼状胬肉的疗效.方法 将58例(62眼)复发性翼状胬肉患者随机分为2组,治疗组(32例,35跟)采用角膜缘于细胞移植联合应用丝裂霉素C;对照组(26例,27眼)单纯采用角膜缘干细胞移植术治疗.术后随访6～30个月并比较疗效.结果 治疗组1眼冉复发,再复发率2.86%;对照组5眼再复发,再复发率18.52%,两组差异具有统计学意义(P<0.05).治疗组角膜创面上皮平均愈合时间为(8.34 ±2.38)d,对照组为(7.21±2.55)d,差异无统计学意义(P>0.05).两组均未发现严重并发症.结论 角膜缘干细胞移植联合应用丝裂霉素C治疗复发性翼状胬肉再复发率低,并发症少,为复发性翼状胬肉安全和有效的治疗方法 .     

诱导多能干细胞培养中饲养层制备的条件优化

目的建立分离培养小鼠胚胎成纤维细胞(Mouse embryonic fibroblast MEF)的体系,探讨诱导多能干细胞(induced pluripotent stem cell,iPSC)培养所需饲养层的最佳制备方法,制备高质量的IPSC饲养层细胞。方法采用胰蛋白酶消化法,将昆明小鼠胚胎制成细胞悬液接种培养。用不同浓度的丝裂霉素C处理MEF细胞制备饲养层,观察处理后的细胞活力及有无再增殖现象,在制备的饲养层细胞上接种IPS细胞,观察各组IPS细胞生长状态。结果采用0.0625%胰蛋白酶消化孕12.5-14.5d雌鼠胚胎组织获得高活力的MEF细胞,用15μg/ml的丝裂霉素C处理后MEF细胞无明显增值,细胞活力在90%以上,其IPS细胞生长状况最好,效果最佳。结论该方法可获得高活力的MEF细胞。15μg/ml的丝裂霉素C处理1.5h后的MEF细胞制成的饲养层最适于IPS细胞生长。     

CF-1小鼠胚胎成纤维细胞饲养层的制备**

背景：小鼠胚胎成纤维细胞作为干细胞生长用饲养层,优于无饲养层,它能够分泌一些既促进干细胞生长又能抑制干细胞分化的因子。 目的：建立CF-1小鼠胚胎成纤维细胞饲养层最佳的分离培养方法,分析丝裂霉素C抑制成纤维细胞增殖的最佳浓度与时间,用于培养诱导多能性干细胞。 方法：取孕10~15 d CF-1小鼠,分离胎鼠原代成纤维细胞,经不同质量浓度(5,10,15,20 mg/L)丝裂霉素C处理不同时间(1,1.5,2,2.5,3 h)制备饲养层,并观察其增殖情况。将人诱导多潜能干细胞或胚胎干细胞在丝裂霉素C处理过的饲养层上培养,观察细胞集落生长情况。 结果与结论：制备CF-1小鼠胚胎成纤维细胞饲养层的最佳胎龄为13.0~14.0 d。丝裂霉素C抑制CF-1小鼠胚胎成纤维细胞增殖的最佳浓度及作用时间为10 mg/L,2.5 h,成纤维细胞饲养层可以维持5~7 d。诱导多能性干细胞与胚胎干细胞在经10 mg/L丝裂霉素C处理2.5 h后饲养层上能发育成典型的“鸟巢”状干细胞集落。 关键词：胚胎成纤维细胞;诱导多潜能分化干细胞;饲养层;丝裂霉素C;细胞集落 doi:10.3969/j.issn.1673-8225.2012.10.017     

丝裂霉素联合自体角膜移植术治疗翼状胬肉50例分析

目的探讨丝裂霉素C联合自体角膜缘干细胞移植治疗翼状胬肉的效果。方法对50例（75）眼翼状胬肉患者行翼状胬肉切除和丝裂霉素C联合自体角膜缘干细胞移植术。结果50例（75眼）翼状胬肉患者随访12-24个月，2例复发。结论翼状胬肉切除和丝裂霉素C联合自体角膜缘干细胞移植术疗效好．可较大地降低胬肉术后的复发率。     

巩膜坏死性炎症的诊断与治疗研究

目的探讨巩膜坏死性炎症的病因、病理改变、诊断和治疗。方法巩膜坏死性炎症患者均详细询问病史及病程,眼科A/B超声和CT检查,查血沉和类风湿因子等。用免疫抑制剂/皮质类固醇及血管扩张剂药物和手术治疗。结果药物治疗12例中6例后部巩膜炎治愈;6例丝裂霉素C所致者治愈5例,1例眼球摘除。13例(18眼)前部和1例后部巩膜坏死者手术清除坏死病灶组织及脓性分泌物,用自体筋膜和结膜掩盖6例9眼,自体阔筋膜修补4例6眼,异体巩膜修补2例2眼,异体角膜修补1例眼,均治愈。结论巩膜坏死性炎症的病因多为免疫系统异常和异物刺激抗原抗体复合物沉积于小血管壁引起血管炎症所致。前部病变病程迁延病灶明显诊断较易,治疗以手术联合药物为主;后部病变易漏诊和误诊,治疗以药物为主。丝裂霉素可致巩膜坏死应注意用药浓度和时间。     

Mutations induced in a shuttle vector plasmid exposed to monofunctionally activated mitomycin C

Reductive activation of mitomycin C leads to its covalent binding to DNA, forming monoadducts and cross-links. The cytotoxicity of mitomycin C has been attributed to cross-link formation, whereas monoadducts are assumed to cause mutagenicity. We have developed a ³²P-postlabeling technique to measure mitomycin C DNA adducts. Using this technique, we have measured monoadduct formation in the shuttle vector plasmid pSP189 and have determined mutations induced by monoadduct formation. The shuttle vector plasmid was incubated with mitomycin C under conditions favoring monofunctional activation of mitomycin C. The plasmid was then replicated in human Ad293 cells, rescued in bacteria, and analyzed for mutations in the supF tRNA gene sequence of pSP189. One major mitomycin C/DNA adduct was observed by ³²P-postla-beling and was characterized as a monoadduct ofguanine. When pSP189 was exposed to monofunctionally activated mitomycin C, increases in adduct levels and mutation frequency were found to be related to mitomycin C concentration. The majority of the mutations involved single bases, with base substitutions making up 59.1% of the total mutations observed. Of the base substitutions, 67.2% were transversions and 32.8% were transitions, with nearly 80% of all base substitutions involving G:C base pairs. Deletions, either as single bases or large deletions, also involved G:C base pairs the majority of the time. The observed bias of mutations atG:C and the formation of a mitomycin C/DNA monoadduct involving guanine suggests that monoadduct formation may be responsible for the mutations. Environ. Mol. Mutagen. 29:143–151, 1997. © 1997 Wiley-Liss, Inc.     

越橘叶黄素酯软胶囊缓解视疲劳的保健作用

目的：观察越橘叶黄素酯软胶囊对缓解视疲劳功能的效果.方法：选择长期用眼,视力易疲劳的成人104例,随机分成试食组和对照组,每组各52例.试食组食用越橘素酯软胶囊,对照组食用安慰剂,食用方法均为每天1次,每次1粒.结果：经过45 d试食后,试食组对眼痛、眼胀、畏光、视物模糊、眼干涩等临床症状有所改善,试食组临床观察总有效率为80.77％,与对照组（23.88％）比较差异有显著性（P＜0.05）.试验组明视持久度较试验前提高0.11,与自身试食前及对照组试验后比较,差异皆有显著性（P＜0.05）.结论：越橘叶黄素酯软胶囊具有缓解视疲劳功能的作用.     

Quantitation of mitomycin C in human ocular tissues by high-performance liquid chromatography-photo-diode array detection

A chromatographic method, which can quantitate mitomycin C (MMC) along with two antiglaucoma drugs, is described. The separation of MMC, alphagan and timolol was performed on a reversed-phase C18 column with water-methanol-trifluoroacetic acid (65:35:0.01, v/v) as the mobile phase. By monitoring at 360, 248 and 296 nm, the lower limits of detection for MMC, alphagan and timolol are, respectively, 1.0, 2.0 and 5.0 ng (injection amount) at three-time S/N ratio. The dynamic ranges of quantitation for the three drugs are, respectively, 1.0 ng-10.0 microg, 2.0 ng-10.0 microg and 5.0 ng-10.0 microg with linearity being larger than 0.9960. This method was applied to the determination of MMC levels in Tenon's and trabeculum tissues of 10 glaucoma patients. MMC levels in these tissues, which were obtained from glaucoma filtering surgery, were determined following a multiple extraction with methanol. The recovery of MMC for a two-batch extraction was better than 91.2%. The reproducibility of measurement for the MMC levels in these tissues is 2.5-6.0% RSD for triplicate injections. The intra-day variation of retention times for the MMC peaks was less than 1.6% RSD (n=3). The inter-day variation of retention times for the MMC peaks was less than 4.8% RSD (n=3). MMC was detectable in three trabeculum tissues out of 10 cases (ranging from 0.8 to 25.5 ng/mg specimen), while MMC was detected in nine Tenon's tissues out of 10 cases (ranging from 0.3 to 21.1 ng/mg specimen). The results obtained show that the method is sensitive and selective for the quantitation of MMC.     

HPLC法测定癌痛消方及其拆方中松脂醇二葡萄糖苷和芍药苷的含量

目的 建立癌痛消方及其拆方中松脂醇二葡萄糖苷和芍药苷的测定方法。方法 采用十八烷基硅烷键合硅胶柱（Inertsil ODS-3柱 250×4.6 mm,5 μm）;流动相:乙腈-0.1%磷酸（15∶85）为流动相,进行梯度洗脱;检测波长:230 nm（0~16 min,松脂醇二葡萄糖苷）,277 nm（16~30 min,芍药苷）;流速:1.0 ml/min;柱温:30oC。结果 癌痛消-补虚组中松脂醇二葡萄糖苷在1.0~5.0 μg范围内呈现良好的线性关系,r=0.9992,平均回收率为97.23%（RSD=1.31%,n=6）;癌痛消-抑癌组中芍药苷在1.0~5.0 μg范围内呈现良好的线性关系,r=0.9993,平均回收率为97.08%（RSD=1.38%,n=6）;癌痛消-全方中松脂醇二葡萄糖苷在1.0~5.0 μg范围内呈现良好的线性关系,r=0.9992,平均回收率为98.01%（RSD=1.30%,n=6）;癌痛消-全方中芍药苷在1.0~5.0μg范围内呈现良好的线性关系,r=0.9993,平均回收率为96.97%（RSD=1.54%,n=6）;结论 本方法专属性强,准确度高,重现性良好,可用于癌痛消方的质量控制。     

An examination of the mitomycin C induction of chromosome polymorphisms in cultures of Plasmodium falciparum.

Cloned parasites of the FCR3 strain of Plasmodium falciparum that survived treatment with either mitomycin C or ethidium bromide were grown and subcloned. The chromosomes of 10 cloned isolates from each population were analysed by contour-clamped homogenous electric field gel electrophoresis. Eight of 10 isolates from the mitomycin C-treated population contained a detectable polymorphic chromosome change while none of the isolates from the ethidium bromide-treated population did. One of the polymorphic changes involved chromosome #4. A pyrimethamine resistant derivative of FCR3, FCR3-D5, that had previously been shown to contain a single nucleotide change in the dihydrofolate reductase-thymidylate synthase (DHFR-TS) gene but no detectable chromosome polymorphisms, was sequentially treated with mitomycin C and a higher concentration of pyrimethamine than the strain had previously been shown to be resistant to in order to determine if there was a correlation between the selection of chromosome #4 polymorphisms and the applied selective pressure. Most of the cloned survivors of this treatment were found to contain chromosome polymorphisms that involved chromosome #4, the chromosome on which the DHFR-TS gene is located. The polymorphic changes were usually different in the different isolates. The selection of mitomycin C-treated, pyrimethamine resistant strains with chromosome #4 polymorphisms will be discussed.     

Combined intravesical hyperthermia and mitomycin chemotherapy: a preliminary in vivo study

Previous clinical studies of the combination of local intravesical hyperthermia with cytostatic drugs for the treatment of Superficial Transitional Cell Carcinoma of the urinary bladder (STCCB) showed encouraging results both in reducing recurrence rate to 20-30% within 2 years and in ablative success rate of 79%. Our objectives were to evaluate bladder tissue and adjacent organs during and following hyperthermia treatment. An intravesical catheter equipped with a radio-frequency antenna (Synergo SB-TS 101.1 System) was used for hyperthermia and intravesical chemotherapy (mitomycin C) was instilled in vivo for 60 min in two anaesthetized sheep. Thirteen to fifteen thermocouples were sewn surgically on the internal and external surfaces of the bladder wall and on adjacent organs to monitor the temperature during the treatment. We expected the intravesical temperature to be under 46 degrees C and the external layers below 45 degrees C. The bladder was filled with 50 mL of chemotherapeutic solution (400 micro g/mL of mitomycin C in distilled water). The sheep were sacrificed at the end of the treatment. Three other sheep, which underwent thoracic surgery, served as control group. Histological changes in both groups showed foci of oedema and haemorrhage with inflammation in the lamina propria and serosa. Foci of desquamation of the epithelium were noticed in the treated sheep. Histological analysis of the treated group showed no significant differences from the control group. The control group showed similar changes, some less pronounced. The combined treatment of hyperthermia with mitomycin C did not cause major damage to the urinary bladder or adjacent organs. All changes were superficial and reversible, and the control group showed similar changes, some less pronounced. Although this is an experimental model based on one single session treatment, rather than repeated treatments, it suggests that the approach may be useful in future studies both in models and man.     

Nd：YAG泪道激光成形术治疗泪道阻塞的体会

目的 探讨应用泪道激光治疗泪道阻塞的方法及疗效。方法 应用Nd：YAG泪道激光仪按泪道探通方法连续击射阻塞部位。用抗生素和丝裂霉素C混合液冲洗畅通后植入义管，每周冲洗一次，1月后拔管。结果共治疗53例60眼，随访1～3月。治愈57眼占95％，好转2眼占3．33％，无效1眼仍有溢脓，1月后行泪囊鼻腔吻合术治愈。结论 激光泪道成形术治疗泪总管、鼻泪管阻塞及泪囊炎，方法简单、疗效满意。     

茴拉西坦及其制剂含量测定方法的改进

目的:改进茴拉西坦及其制剂的含量测定方法。方法:采用高效液相色谱法,以C18为色谱柱,流动相为乙腈-水(35:65),检测波长283nm。结果:茴拉西坦在0.07792～6.2336μg范围内,线性关系良好(r=1.0000);平均回收率为100.2%,RSD为1.6%。结论:方法准确可靠,结果重现性好,可用于茴拉西坦的质量控制。     

Mitomycin C associated hemolytic uremic syndrome

Mitomycin C associated Hemolytic Uremic Syndrome (HUS) is a potentially fatal but uncommon condition that is not yet widely recognised. It consists of microangiopathic hemolytic anemia, thrombocytopenia and progressive renal failure associated with mitomycin C treatment and affects about 10% of patients treated with this agent. The renal failure usually develops about 8-10 mth after start of mitomycin C treatment and the mortality is approximately 60% from renal failure or pulmonary edema. Renal lesions are similar to those seen in idiopathic HUS and include arteriolar fibrin thrombi, expanded subendothelial zones in glomerular capillary walls, ischemic wrinkling of glomerular basement membranes and mesangiolysis. The mechanism of action is postulated as mitomycin C-induced endothelial cell damage. We describe the clinical course and pathological findings in a 65 yr-old man with gastric adenocarcinoma who developed renal failure and thrombocytopenia while on treatment with mitomycin C and died in pulmonary edema.