3'-Azido-2',3'-dideoxythymidine induced deficiency of thymidine kinases 1, 2 and deoxycytidine kinase in H9 T-lymphoid cells

Continuous cultivation of T-lymphoid H9 cells in the presence of 3′-azido-2′,3′-dideoxythymidine (AZT) resulted in a cell variant cross-resistant to both thymidine and deoxycytidine analogs. Cytotoxic effects of AZT, 2′,3′-didehydro-3′-deoxythymidine as well as different deoxycytidine analogs such as 2′,3′-dideoxycytidine, 2′,2′-difluoro-2′-deoxycytidine (dFdC) and 1-ß-D-arabinofuranosylcytosine (Ara-C) were strongly reduced in H9 cells continuously exposed to AZT when compared to parental cells (>8.3-, >6.6-, >9.1-, 5×10⁴-, 5×10³-fold, respectively). Moreover, anti-HIV-1 effects of AZT, d4T, ddC and 2′,3′-dideoxy-3′-thiacytidine (3TC) were significantly diminished (>222-, >25-, >400-, >200-fold, respectively) in AZT-resistant H9 cells. Study of cellular mechanisms responsible for cross-resistance to pyrimidine analogs in AZT-resistant H9 cells revealed decreased mRNA levels of thymidine kinase 1 (TK1) and lack of deoxycytidine kinase (dCK) mRNA expression. The loss of dCK gene expression was confirmed by western blot analysis of dCK protein as well as dCK enzyme activity assay. Moreover, enzyme activity of TK1 and TK2 was reduced in AZT-resistant cells. In order to determine whether lack of dCK affected the formation of the active triphosphate of the deoxycytidine analog dFdC, dFdCTP accumulation and retention was measured in H9 parental and AZT-resistant cells after exposure to 1 and 10 μM dFdC. Parental H9 cells accumulated about 30 and 100 pmol dFdCTP/10⁶ cells after 4 hr, whereas in AZT-resistant cells no dFdCTP accumulation was detected. These results demonstrate that continuous treatment of H9 cells in the presence of AZT selected for a thymidine analog resistant cell variant with cross-resistance to deoxycytidine analogs, due to deficiency in TK1, TK2, and dCK.     

补肾健脾养血活血方对女性血管性痴呆患者性激素和血脂水平的影响

目的：研究补肾健脾养血活血方对女性血管性痴呆（VD）患者血清雌二醇（E2）、睾酮（T）、总胆固醇（TC）、甘油三脂（TG）、高密度脂蛋白（HDL）及低密度脂蛋白（LDL）水平的影响。方法：选取女性VD患者31例,将其随机分为中药治疗组（治疗组）16例,西药对照组（对照组）15例,治疗组给予由补肾健脾养血活血方制成的复方胶囊制剂,对照组予以西药喜得镇治疗。两组患者均治疗1个月为1个疗程,连服3个疗程。同时选取20例正常人群作为正常组,进行疗效比较。结果：①治疗组有效9例,显效5例,总有效率为87.5%;对照组有效6例,显效4例,总有效率为62.5%。两组总有效率比较有显著性差异。30例女性VD患者与正常组比较,血清E2水平降低（P〈0.05）,TC、TG、LDL水平提高（P〈0.05）,两组间比较均有显著性差异。②治疗组E2、E2/T、TC、TG、HDL与本组治疗前比较有显著性差异（P〈0.05或P〈0.01）,治疗后E2、T、E2/T、TC、TG、HDL与对照组比较有显著性差异（P〈0.05或P〈0.01）。结论：补肾健脾养血活血方可通过补肾调节性激素及血脂水平,对女性VD患者有一定疗效,是防治VD的有效方药。     

ISO/TC215传统医学信息标准跟踪研究

调研了ISO／TC215健康信息学技术委员会的传统医学信息标准化工作,介绍了“中医药语苦系统的语义网络框架”、“中医药文献元数据”、“传统医学中临床所见的描述结构”、“草药术语系统的分类结构”等4个ISO／TO 215正在进行的标准项目,可供传统医学信息标准研究人员参考。     

Aqueous extract of unripe Rubus coreanus fruit attenuates atherosclerosis by improving blood lipid profile and inhibiting NF-κB activation via phase II gene expression

Ethnopharmacological relevance

The fruit of Rubus coreanus has been used as a traditional herbal medicine for alleviation of inflammatory and vascular diseases in Asian countries.

Aim of the study

The anti-atherogenic effect of unripe Rubus coreanus fruit extract (URFE) and its underlying mechanism were analyzed in mice fed a high-fat diet (HFD) and in cell culture system.

Materials and methods

Mouse was freely given HFD alone or supplemented with URFE for 14 weeks, followed by analysis of atherosclerotic lesions and serum lipid levels. For in vitro assay, macrophages were pretreated with URFE, followed by stimulation with lipopolysaccharide (LPS). Expression levels of inflammatory genes (TNF-α, IL-1β, and iNOS) and phase II genes (heme oxygenase-1, glutamate cysteine lygase, and peroxiredoxine-1) as well as intracellular reactive oxygen species (ROS) level and NF-κB activation pathway were analyzed in cultured macrophages as well as mouse sera and aortic tissues.

Results

URFE supplementation reduced HFD-induced atherosclerotic lesion formation which was correlated with decreased levels of lipids, lipid peroxides, and inflammatory mediators (TNF-α, IL-1β, and nitric oxide) in sera as well as suppression of inflammatory gene in aortic tissues. In addition, pre-treatment of macrophages with URFE also suppressed LPS-induced NF-κB activation, ROS production, and inflammatory and phase II gene expressions. Inhibition of phase II enzyme and protein activities attenuated the suppressive effects URFE on ROS production, NF-κB activation, and inflammatory gene expression.

Conclusion

These results suggest that URFE attenuates atherosclerosis by improving blood lipid profile and inhibiting NF-κB activation via phase II antioxidant gene expression.     

The p.R1109X mutation in SH3TC2 gene is predominant in Spanish Gypsies with Charcot-Marie-Tooth disease type 4

Charcot-Marie-Tooth (CMT) disease type 4 (CMT4) is the name given to autosomal recessive forms of hereditary motor and sensory neuropathy (HMSN). When we began this study, three genes or loci associated with inherited peripheral neuropathies had already been identified in the European Gypsy population: HMSN-Lom (MIM 601455), HMSN-Russe (MIM 605285) and the congenital cataracts facial dysmorphism neuropathy syndrome (MIM 604168). We have carried out genetic analyses in a series of 20 Spanish Gypsy families diagnosed with a demyelinating CMT disease compatible with an autosomal recessive trait. We found the p.R148X mutation in the N-myc downstream-regulated gene 1 gene to be responsible for the HMSN-Lom in four families and also possible linkage to the HMSN-Russe locus in three others. We have also studied the CMT4C locus because of the clinical similarities and showed that in 10 families, the disease is caused by mutations located on the SH3 domain and tetratricopeptide repeats 2 (SH3TC2) gene: p.R1109X in 20 out of 21 chromosomes and p.C737_P738delinsX in only one chromosome. Moreover, the SH3TC2 p.R1109X mutation is associated with a conserved haplotype and, therefore, may be a private founder mutation for the Gypsy population. Estimation of the allelic age revealed that the SH3TC2 p.R1109X mutation may have arisen about 225 years ago, probably as the consequence of a bottleneck.     

The involvement of NR4A1 and NR4A2 in the regulation of the luteal function in rats

The nuclear receptor 4A (NR4A) members play important roles in cellular proliferation, differentiation and apoptosis. The current study first evaluate the expression of ovarian NR4A1 during different luteal stages in rats. Immature rats aged 28 days were treated with sequential Pregnant mare serum gonadotropin (PMSG) (D -2) / human chorionic gonadotropin (hCG) (D 0) to induce pseudopregnancy. Serum progesterone (P₄) and ovarian expression of NR4A1 were detected by RIA and WB, respectively, at follicle stage (D 0), early (D 2), middle (D 7) and late (D 14 and D 20) luteal stages. To confirm the role of NR4A1 during the luteal regression, rats were treated with prostaglandin F_2α analog (PGF) for 0–8?h on D 7 to detect the expressions of NR4A1 and NR4A2. RIA result showed that serum P₄ reached highest level on D 7 and then declined. WB results showed that there were two types of NR4A1 (NR4A1-L and NR4A1-S) expressed in the ovary. The ovarian NR4A1-L decreased at the late luteal stage (D 20). However, the NR4A1-S increased at the late luteal stage (D 14). After PGF treatment on D 7, the expression of NR4A1-S increased which peaked at 0.5–1?h and then declined; while NR4A1-L expression did not change within 8 h. Real-time PCR results showed that the ovarian NR4A1 mRNA increased within 0.5?h, maintained high at 1 h and then declined. The NR4A2 mRNA expression exhibited a similar pattern to that of NR4A1 mRNA, though its abundance was not as high as NR4A1. IHC results revealed that NR4A1-L was expressed mainly in the cytoplasm of luteal steroidogenic cells, faintly expressed in the follicle theca cells, oocytes and the pericytes; while NR4A2 was primarily localized in the cytoplasm of luteal steroidogenic cells. In conclusion, all these results demonstrate that NR4A2 as well as NR4A1 might be involved in the luteal development and luteolysis in rats.     

常州市成人血脂异常的流行特征分析

目的探讨常州地区当前一般人群血脂异常的流行特点。方法利用2012年1 259名健康检查者资料,分析并比较不同性别、不同年龄人群血清甘油三酯（TG）、总胆固醇（TC）、高密度脂蛋白胆固醇（HDL-C）、低密度脂蛋白胆固醇（LDL-C）的异常率以及TG与TC、LDL-C、HDL-C的关系。结果①男性TG、TC、LDL-C、HDL-C异常率均高于女性,按年龄分层,男女TC、LDL-C异常率均随年龄增加呈上升趋势,但男性TG异常率与年龄不呈正比。无论男性还是女性,不同年龄段HDL-C异常率未见统计学差别。②男性血脂异常者中有49.9%为混合型,女性则有52.2%为混合型。③logistic回归分析表明,校正性别、年龄后,TG异常与TC异常呈正相关（OR 95%CI=1.563~3.409）,与HDL-C异常呈负相关（OR 95%CI=0.246~0.974）。结论常州地区成年人血脂异常率较高,且混合型血脂异常占较高比例,今后应积极开展预防血脂异常的健康教育。