参附注射液、高乌甲素注射液诱导白血病细胞株HL-60分化、凋亡的研究

【目的】观察参附注射液及高乌甲素注射液诱导白血病细胞株HL-60分化、凋亡的作用,初步验证温阳法治疗肿瘤的理论。【方法】采用人类急性白血病细胞株HL-60为模型,设参附注射液组(终浓度分别为12.5、25、50、100μL/mL),高乌甲素组(终浓度分别为25、50、100μL/mL),亚砷酸钠对照组(终浓度分别为0.25、2、15μmol/L)及阴性对照组;检测细胞增殖、细胞形态、硝基四氮唑蓝(NBT)还原能力、细胞周期分布、凋亡率等指标。【结果】参附注射液作用后HL-60细胞增殖受抑制,高乌甲素注射液对HL-60的生长无抑制作用;经药物处理60 h后HL-60细胞均出现程度不同以凋亡为主的形态改变;低浓度参附注射液(12.5μL/mL)、高乌甲素注射液(50μL/mL)作用60 h后HL-60细胞NBT还原能力增强,而其他浓度组NBT还原能力反未增强;药物处理60 h后,100μL/mL高乌甲素液及25.50μL/mL参附液流式细胞仪检测直方图上呈现亚二倍体凋亡峰,凋亡率呈不同程度增高。【结论】参附注射液、高乌甲素注射液对人类急性白血病细胞株HL-60具有不同程度的诱导分化和凋亡的双重作用,为温阳法治疗肿瘤提供了一定的理论依据。

HPLC法测定注射用氢溴酸高乌甲素含量的研究

目的建立测定注射用氢溴酸高鸟甲素含量的高效液相色谱法。方法色谱条件：以十八烷基硅烷键合硅胶为填充剂；以0·025moi·L^-1磷酸二氢钾-甲醇（52：48），并用磷酸调pH值至4．0为流动相；检测波长为252nm。结果线性范国：20·72-207.20μg·ml^-1，相关系数r=0．9996．平均回收率为99．6％，RSD为0．42％，高鸟甲素及其有关杂质得到界限分离。结论本方法快速灵敏，准确实用，适用于注射用氢溴酸高鸟甲素的含量测定。

酮咯酸氨丁三醇与高乌甲素在脊柱外科术后镇痛的临床应用

目的比较脊柱外科术后单纯应用芬太尼与酮咯酸氨丁三醇或高乌甲素复合芬太尼静脉镇痛的效果和不良反应。方法选择脊柱外科手术患者60例，ASAⅠ～Ⅱ级，随机分成3组，每组20例。3组均采用异丙酚、芬太尼全凭静脉麻醉。其中A组术后单纯芬太尼1．6mg静脉镇痛；B组于麻醉诱导前静脉给予酮咯酸氨丁三醇30mg，术后芬太尼0．6mg，酮咯酸氨丁三醇90mg静脉镇痛；C组于麻醉诱导前给予高乌甲素8mg作为负荷量，术后芬太尼O．6mg，高乌甲素20nag静脉镇痛。分别观察3组术后镇痛效果和不良反应。结果3组镇痛效果均良好，A组优于B、C组，B组优于C组（P〈0．05）。B组与C组嗜睡、过度镇静、恶心呕吐和轻度低氧血症（SpO2〈95％）的发生率明显低于A组（P〈0．05），但C组高血压、窦速发生率明显高于A组（P〈0．05）。结论在脊柱外科手术后复合应用酮咯酸氨丁三醇或高乌甲素，可以显著减少芬太尼用量，达到良好的术后镇痛效果。使术后镇痛更加舒适而不良反应少。

鸭病毒性肝炎的中西医结合治疗

鸭病毒性肝炎是鸭肝炎病毒引起的雏鸭高度致死性的急性传染病。主要是危害3周龄以内的雏鸭。鸭肝炎表现为发病突然,病程短促、食欲废绝、运动失调、倒地呈游泳状,死后角弓反张。鸭病毒性肝炎发病率和死亡率都很高,是目前养禽业常见的病毒性疾病之一。介绍该病的中西医治疗方法。

Weekly low-dose Semustine in a patient with peripheral T-cell lymphoma,not otherwise specified (PTCL-NOS) coupled with subcutaneous involvement and positive O^6-methylguanine-DNA methyltransferase (MGMT) promoter methylation

Peripheral T-cell lymphoma, not otherwise specified （PTCL-NOS） is a heterogeneous group of aggressive T-ce lymphomas with no treatment consensus and poor prognosis. We herein described an extraordinary refractory case of PTCL-NOS with widely involvement of subcutaneous tissue, which showed an excellent response to Semustine personal- ized chemotherapy based on the detection finding of positive O6-methylguanine-DNA methyltransferase （MGMT） promoter methylation. This is the first english report to indicate that single nitrosourea agent such as Semustine may have good efficacy and safety for widespread subcutaneous involvement of PTCL-NOS with positive MGMT promoter methylation.

Temozolomide and radiotherapy in newly diagnosed glioblastoma patients:O^6-methylguanine-DNA methyltransferase (MGMT) promotor methylation status and Ki-67 as biomarkers for survival and response to treatment

Objective:This phase II study aimed at investigating the correlation between O6-methylguanine-DNA methyltransferase (MGMT) promoter methylation and protein expression,together with Ki-67 labeling index (LI),to response,time to progression (TTP),and overall survival (OS) in newly diagnosed glioblastoma multiforme (GBM) patients treated with temozolomide (TMZ) concomitant with and adjuvant to radiotherapy (RT).Methods:From June 2005 to August 2008,34 adult patients (18-65 years),PS ≥70,with newly diagnosed GBM received TMZ 75 mg/m2 plus RT up to 60 Gy,followed by TMZ 175 mg/m2 5 days every 4 weeks for 12 doses.MGMT Methylation-specific PCR assay,MGMT protein expression,and Ki-67 expression using immunohistochemistry (IHC) were performed on the tissue blocks.The patients were followed by MRI while MR spectroscopy (MRS) was performed for the stable cases or to confirm progression and accordingly Bevacizumab 10 mg/kg every 2 weeks was added to 7 patients till further progression was proved.Results:31 cases were evaluable,12 (38.7%) had unmethylated MGMT,while 19 (61.3%) were methylated.Seventeen cases (55%) were MGMT immunonegative while 14 cases (45%) were immunopositive.The cut off value of Ki-67 LI in relation to survival was 17%,where 15 were < 17% (48.4%),and 16 were ≥ 17% (51.6%).Response evaluation started after the second dose of the adjuvant TMZ and was repeated every 2 months.The overall disease control rate (ODC) was 74.2%,where 2 patients had complete response (CR),14 had partial response (PR),and 7 had stable disease (SD),while 8 (25.8%) had progressive disease (PD).The ODC was significantly higher among methylated patients and in those with Ki-67 < 17% (P=0.0003).The median overall TTP was 12 months and the median OS was 20 months for all the patients including those who received Bevacizumab for some stable cases or as a salvage treatment in patients with good PS,the MGMT-methylated patients had a higher median TTP of 13 months (range 8 to 18 months,95% CI of 9.36 to 12.9),and OS of 24 months (range 12 to 31 months,95% CI of 16.1 to 21.32),while the unmethylated patients had a median TTP of 6.5 months and a median OS of 12 months,such correlations were highly significant (P=0.0001).MGMT immunoexpression failed to show significant correlation with MGMT promotor methylation or the outcome of the patients.Patients with Ki-67 < 17% had a median TTP of 16 months and median OS of 24 months compared to 7 and 12.5 months respectively for the patients with Ki-67 ≥17%.Significant correlation was found between the ODC,TTP,and OS with age < 52,near total excision,and TMZ doses received ≥ 10.The commonest grade 3 and 4 toxicities was neutropenia recorded in 3 patients (9.67%),thrombocytopenia in 4 patients (12.9%),and one patient with G3 nausea,vomiting,and constipations (3%),all were medically manageable.Conclusion:MGMT promotor methylation status and Ki-67 LI (but not the MGMT protein expression),could serve as prognostic markers for survival,also MGMT could identify the newly diagnosed GBM patients who will have better response to TMZ.

Guanine-regulated proton transfer enhances CO_(2)-to-CH_(4) selectivity over copper electrode

Electrocatalytic CO_(2) reduction has attracted growing attention as a promising route to realize artificial carbon recycling.Proton transfer plays an essential role in CO_(2) reduction and dramatically impacts product distribution.However,the precise control of proton transfer during CO_(2) reduction remains challenging.In this study,we present a well-controlled proton transfer through the modification of several purines with similar molecular structures,and reveal a direct correlation between surface proton transfer capability and CO_(2) reduction selectivity over Cu electrode.With a moderate proton transfer capability,the guanine modification can remarkably boost CH_(4) production and suppress C2 products formation.In-situ ATR-SEIRAS suggests a weakened^(*)CO intermediate adsorption and a relatively low local pH environment after the guanine modification,which facilitates the^(*)CO protonation and detachment for CH_(4) generation.

Ecological Research of Former Brown-coal Quarry-the Most Lake in the Czech Republic

The presented article discusses the results of survey of the Most lake, which was created within hydric recultivation of the site of the former brown-coal mine Most Lezaky in northern Bohemia in the Czech Republic. The survey discovered plants which belong to ruderal, wetland, halophilic and xerothermal species based on their life strategy. The discovered endangered species include Carex secalina, Schoenoplectus tabaernemontani, Salsola call subsp. Rosacea and Tetragonolobus maritimus. A total of 350 species of vascular plants were discovered in the study area. During the monitoring, a total of 146 bird species were discovered. The monitored location can be currently considered as a significant gathering place namely for aquatic and wetland bird species during the winter period and the migration period （e.g., Tachybaptus ruficollis, Podiceps cristatus, Anser anser, Anas platyrhynchos, Anas strepera, Fulica atra and various species of Larus sp.） and as a significant area of occurrence of relatively rare species bound to anthropogenetically disturbed areas of mining and post-mining landscape （e.g., Anthus campestris, Anthus pratensis, Motacillaflava, Oenanthe oenanthe, Saxicola rubetra and Saxicola torquata）. Within the monitoring of the water quality, the saprobic index values （S） ranged in 2013 between 1.62 and 1.92.

Plating densities, alpha-difluoromethylornithine effects and time dependence on the proliferation of IEC-6 cells

OBJECTIVE: To characterize the role of plating densities and alpha-difluoromethylornithine (DFMO) on the proliferation of IEC-6 cells in vitro. METHODS: IEC-6 cells were seeded in 96-well microplates at various densities in the presence or absence of DFMO. Cells were counted and their proliferative capability was monitored Days 1 to 7 with MTT assay at an optical density of 570 nm. RESULTS: There was a positive relationship between cell number and OD value (r = 0.954, P 0.5 x 10(4) cells/well) inhibited the growth of cells on Day 2. When the density reaches 4 x 10(4) cells/well, the OD value increased gradually and reached a peak on Day 5. After that, the OD value began to fall. The growth of IEC-6 cells was limited at a low density (0.2 x 10(4) cells/well) on Day 4. DFMO caused a complete inhibition of proliferation of IEC-6 cells on Days 1 to 3. CONCLUSION: Proliferation of IEC-6 cells is related to plating density and incubation time. It is inhibited by DFMO, but is reversible when the incubation time is prolonged.

Plating densities, alpha-difluoromethylornithine effects and time dependence on the proliferation of IEC-6 cells

To characterize the role of plating densities and alpha difluoromethylornithine (DFMO) on the proliferation of IEC 6 cells in vitro Methods IEC 6 cells were seeded in 96 well microplates at various densities in the pre sence or absence of DFMO Cells were counted and their proliferative capabilit y was monitored Days 1 to 7 with MTT assay at an optical density of 570?nm Results There was a positive relationship between cell number and OD value ( r =0 954 , P 【0 01) Higher plating densities (】0 5×10 4 cells/well) inhibited the growth of cells on Day 2 When the density reaches 4×10 4 cells/well, the O D value increased gradually and reached a peak on Day 5 After that, the OD va lue began to fall The growth of IEC 6 cells was limited at a low density (0 2×10 4 cells/well) on Day 4 DFMO caused a complete inhibition of proliferati on of IEC 6 cells on Days 1 to 3 Conclusion Proliferation of IEC 6 cells is related to plating density and incubation time It is inhibited by DFMO, but is reversible when the incubation time is prolon ged

1-Methyl-4-phenyl-pyridinium time-dependently alters expressions of oxoguanine glycosylase 1 and xeroderma pigmentosum group F protein in PC12 cells

Objective To determine if DNA excision repair enzymes oxoguanine glycosylase 1 （OGG1） and xeroderma pigmentosum group F protein （XPF） are involved in the pathogenesis of Parkinson＇s disease （PD） in a cell model. Methods PC12 cells were treated with 1-Methyl-4-phenylpyridine ion （MPP＋） for various periods of time to induce oxidative DNA damage. MTT assay was used to determine cell viability. Immunocytochemistry with antibody against 8-hydroxy-2＇- deoxyguanosine （8-oxodG） was used to evaluate oxidative DNA damage. Immunoblotting was used to detect the protein levels of OGG1 and XPF. Results MPP＋ treatment （1 mmol/L） for 18 h and 24 h reduced cell viability to 78.6% and 70.3% of the control, respectively, in a time-dependent way. MPP＋ increased the immunoreactivity of 8-oxodG in the cytoplasm at 3 h and in the nucleus at 24 h of treatment. With the treatment of MPP＋, the expression of OGG1 was significantly increased at 1 h, reaching a peak at 3 h, and then it was decreased at 24 h, as compared to that with vehicle treatment. The same effect was exerted on XPF level, except that the XPF level reached a peak at 18 h of MPP＋ treatment. Moreover, the maximally-increased protein level of OGG1 by MPP＋ was approximately 2-fold higher than that of XPF. Conclusion MPP＋ treatment could time- dependently induce increases in OGG1 and XPF expressions in PC12 cells. Also, this study indicates that the base and nucleotide excision repair pathways may be compensatorily activated in the early stage of pathogenesis in the cells after MPP＋ treatment.

Analysis of treatment regimen in a pregnant patient with ornithine transcarbamylase deficiency

As a common urea-cycle disorder, ornithine transcarbamylase deficiency(OTCD) disables the conversion of ammonia into non-toxic urea, and its subsequent excretion results in hyperammonemia. We reported a 28-year-old woman who was diagnosed with OTCD during her first pregnancy. She was treated with arginine, citrulline and sodium benzoate for complications associated with hyperammonemia, and her protein intake was restricted. The patient’s condition was stabilized, and she delivered a baby boy via cesarean section. However, the baby died 2d later. During her second pregnancy, prenatal screening suggested that the fetus had OTCD, and an induced abortion was performed. During her third pregnancy, fetal OTCD was ruled out, and the patient was treated with oral sodium benzoate. Her blood ammonia level was stabilized, and a baby boy was successfully delivered via cesarean section. This case described the treatment process of the pregnant patient with OTCD, and the safety and efficacy of sodium benzoate were evaluated. Collectively, our findings provided the experience and evidence for the drug selection and treatment of these rare diseases.