Metabolism of free fatty acids（FFAs） is related to several important physiological events and therefore their quantitaion in biological samples arouses extensive interest and efforts.Existing gas chromatography with...Metabolism of free fatty acids（FFAs） is related to several important physiological events and therefore their quantitaion in biological samples arouses extensive interest and efforts.Existing gas chromatography with flame ionization detector（GC-FID） methods for the analysis of FFAs normally require derivatization of them in order to lower boiling points.But this extra procedure tends to induce additional error and it is laborious and time-consuming.A derivatization-free method was therefore established in the present investigation to determine FFAs in human plasma by capillary（GC-FID）.After extraction of FFAs from plasma,a highly polar FFAP（free fatty acid in plasma） column was employed to directly quantitate FFAs concentration,free from derivatization reaction.All sample pretreatments were carried out at room temperature,improving recovery of short-chain FFAs.Heptadecanoic acid（C17：0） was employed as internal standard,and the proposed method was validated for recovery,precision,sensitivity,stabi-lity,and linearity.Validation data show that it is suitable for clinical study that has been applied to the evaluation of FFAs levels in plasma of diabetic nephropathy（DN） patients during the course of treatment.Forty-seven patients diagnosed with DN were admitted to the double-blind experiment.Control group（n=17） underwent solely basic treatment and the patients did not show significant change in FFAs concentration during six months of treatment.Experiment group（n=30） was supplied with traditional Chinese medicine besides basic treatment.After six months of medication,their plasma concentration of palmitic acid（C16：0）,stearic acid（C18：0） and oleic acid（C18：1n-9） decreased while linolenic acid（C18：3n-3） increased significantly（P〈0.05）.These four compounds could be served as biomar-kers in the evaluation of drug efficacy,and their quantitation in plasma may provide additional information for disease progression in DN patients.展开更多
Objective To investigate the therapeutic effect of intensive glycemic control on patients with early diabetic nephropathy. Methods A total of 41 type 2 diabetes patients who developed microalbuminuria were divided int...Objective To investigate the therapeutic effect of intensive glycemic control on patients with early diabetic nephropathy. Methods A total of 41 type 2 diabetes patients who developed microalbuminuria were divided into two groups randomly. Patients in Group A received intensive glycemic control and the blood glucose in Group B was regularly controlled. Glycemic monitoring and control were followed for 12 weeks to observe the changes of microalbuminuria in both groups; meanwhile the levels of serum lipids and coagulation indices were also recorded. Results The urine albumin excretion rate (UAER) in Group A decreased significantly from (47.91±13.86)mg/24h to (35.31±14.56)mg/24h after 12 weeks (P<0.05),and this decrease was significantly greater than that in Group B. However,Group B had no significant difference in UAER decrease [(48.93±13.32)mg/24h to (40.48±19.62)mg/24h,P>0.05]. The decrease of triglyceride (TG) and low-density lipoprotein cholesterol (LDL cholesterol),and the increase of high-density lipoprotein cholesterol (HDL cholesterol) showed no significant differences (P>0.05). And the level of plasma fibrinogen (FIB) showed no significant decrease after 12 weeks,either (P>0.05). Conclusion Intensive glycemic control reduces the level of microalbuminuria and may ameliorate the progression of early diabetic nephropathy.展开更多
Background: Diabetes mellitus (DM) is the leading cause of end stage renal disease (ESRD) worldwide. Although DM with proteinuria is the ultimate result of diabetic nephropathy (DN), a wide spectrum of non-diabetic re...Background: Diabetes mellitus (DM) is the leading cause of end stage renal disease (ESRD) worldwide. Although DM with proteinuria is the ultimate result of diabetic nephropathy (DN), a wide spectrum of non-diabetic renal diseases (NDRD) can occur in such patients. Objective: To observe the frequency and histological pattern of NDRD in diabetic patients with proteinuria and to explore their association with clinical and laboratory parameters. Methods: This cross-sectional study was conducted in the Department of Nephrology, Bangabandhu Sheikh Mujib Medical University (BSMMU), Dhaka, Bangladesh from April 2016 to September 2017. In this study a total of 38 cases of DM with proteinuria (>1 gm/24-hour) were selected purposively. Renal biopsy was done in all patients. Based on histological findings they were categorized into two groups;Group 1 with NDRD and Group II with DN. Their clinical and laboratory parameters were analyzed and compared. Results: Among the total study subjects, 21 (55.3%) were male and 17 (44.7%) were female, mean (±SD) age 43.45 ± 9.99 years in the NDRD group and 41.57 ± 9.50 years in the DN group. Thirty one cases (81.6%) out of thirty eight had NDRD and seven (18.4%) cases had isolated DN;therefore more than two third cases had NDRD. Duration of DM was found to be significantly shorter (p = 0.004) in the NDRD group. Diabetic retinopathy was present in 12.9% cases in NDRD group vs. 57.1% cases in DN group (p = 0.025). Frequency of microscopic hematuria was significantly higher (90.3%) in NDRD patients (p = 0.002). Conclusion: The frequency of NDRD in type 2 diabetic patients other than diabetic nephropathy is relatively high. Membrano proliferative glomeru-lonephritis and membranous nephropathy are more common in NDRD. Absence of diabetic retinopathy, presence of hematuria and shorter duration of DM are markers associated with NDRD in type 2 DM, which are important indicators for renal biopsy in diabetic patients with proteinuria.展开更多
【目的】观察苦瓜皂苷G对糖尿病肾病(DN)大鼠的治疗作用及机制。【方法】从55只大鼠中随机抽取45只采用一次性腹腔注射链脲佐菌素(STZ)法诱导DN模型。将造模成功的38只大鼠随机分为模型组9只、苦瓜皂苷G低剂量组9只、苦瓜皂苷G中剂量组1...【目的】观察苦瓜皂苷G对糖尿病肾病(DN)大鼠的治疗作用及机制。【方法】从55只大鼠中随机抽取45只采用一次性腹腔注射链脲佐菌素(STZ)法诱导DN模型。将造模成功的38只大鼠随机分为模型组9只、苦瓜皂苷G低剂量组9只、苦瓜皂苷G中剂量组10只、苦瓜皂苷G高剂量组10只。剩余10只大鼠设为正常组。对应灌胃给药4周后,酶联免疫吸附分析(ELISA)检测大鼠24 h尿蛋白水平,血糖仪检测空腹血糖(FBG),全自动生化分析仪检测血肌酐(SCr)、血尿素氮(BUN)水平,苏木素-伊红染色(HE)法观察大鼠肾组织病理学变化,采用实时定量聚合酶链反应(RT-qPCR)和Western Blot法分别检测E-钙黏蛋白(E-cad)、α-平滑肌肌动蛋白(α-SMA)、Notch1、锌指蛋白转录因子1(Snail1)、Jagged1 m RNA及蛋白表达情况。【结果】与正常组比较,模型组24 h尿蛋白、FBG、血清SCr和BUN水平及肾组织α-SMA、Notch1、Snail1、Jagged1 mRNA及蛋白水平显著升高,肾组织E-cad m RNA及蛋白水平降低,肾组织可见肾小球萎缩、肾小管扩张及间质纤维化;与模型组比较,苦瓜皂苷G低、中、高剂量组24 h尿蛋白、FBG、血清SCr和BUN水平及肾组织α-SMA、Notch1、Snail1、Jagged1 mRNA及蛋白水平显著降低,肾组织E-cad m RNA及蛋白水平升高,肾组织病理损伤得到改善。【结论】苦瓜皂苷G可有效改善DN大鼠肾功能障碍、减轻肾脏纤维化,其机制可能与通过调控Notch/Snail1信号通路,抑制α-SMA和Jagged1 mRNA及蛋白表达,增强E-cad mRNA及蛋白表达有关。展开更多
目的·探讨甲基转移酶3(methyltransferase like 3,METTL3)调控pri-miR-21的N^(6)-甲基腺苷(N^(6)-methyladenosine,m^(6)A)甲基化修饰在糖尿病肾病(diabetic nephropathy,DN)小鼠肾脏纤维化发病机制中的作用。方法·采用8周龄...目的·探讨甲基转移酶3(methyltransferase like 3,METTL3)调控pri-miR-21的N^(6)-甲基腺苷(N^(6)-methyladenosine,m^(6)A)甲基化修饰在糖尿病肾病(diabetic nephropathy,DN)小鼠肾脏纤维化发病机制中的作用。方法·采用8周龄雄性db/db小鼠作为DN模型小鼠,db/m小鼠作为对照,同时按照是否经尾静脉注射S-腺苷高半胱氨酸水解酶抑制剂3-脱氮腺苷(3-deazaadenosine,DAA),共随机分为4组(5只/组),分别为db/m组、db/db组、db/m+DAA组和db/db+DAA组;8周龄开始注射DAA,注射1次/5 d,共注射8次。DAA干预结束后继续饲养小鼠至19周龄,收取各组小鼠血、尿、肾脏组织标本。检测血糖、血肌酐、尿白蛋白肌酐比(albumin-to-creatinine ratio,ACR),肾脏行苏木精-伊红(H-E)染色、Masson染色及天狼星红染色观察病理变化;试剂盒检测肾脏总RNA中m^(6)A的甲基化水平;Western blotting检测肾脏METTL3及纤维化相关蛋白表达;实时定量PCR检测肾脏总pri-miR-21和成熟miR-21;使用免疫磁珠富集肾脏组织中m^(6)A甲基化RNA,并通过PCR检测其中m^(6)A甲基化的pri-miR-21。结果·相较于db/m组,db/db组小鼠血糖,血肌酐,ACR,肾脏METTL3、m^(6)A甲基化修饰水平、纤维化相关蛋白、总pri-miR-21、m^(6)A甲基化pri-miR-21和成熟miR-21表达水平均显著增加(均P<0.05),小鼠肾脏系膜基质增多、肾小球基底膜增厚、胶原纤维累积显著增加。相较于db/db组,db/db+DAA组血糖,血肌酐,ACR,肾脏m^(6)A甲基化修饰水平、纤维化相关蛋白、m^(6)A甲基化pri-miR-21和成熟miR-21表达水平均显著下降(均P<0.05),总pri-miR-21表达水平显著升高(P=0.000),METTL3蛋白表达水平未见显著变化,小鼠肾脏损伤及纤维化程度显著减轻。结论·pri-miR-21的m^(6)A甲基化修饰促进miR-21成熟,进而促进DN小鼠肾脏纤维化的发生发展;抑制METTL3可通过调控pri-miR-21的m^(6)A甲基化修饰抑制DN小鼠肾脏纤维化。展开更多
基金Supported by the National Basic Research Program of China(Nos.2007CB511903,2005CB523503)the International Cooperation Project of Ministry of Science and Technology of China(No.S2010GR0583)the National Natural Science Founda- tion of China(Nos.90709045,20805026)
文摘Metabolism of free fatty acids（FFAs） is related to several important physiological events and therefore their quantitaion in biological samples arouses extensive interest and efforts.Existing gas chromatography with flame ionization detector（GC-FID） methods for the analysis of FFAs normally require derivatization of them in order to lower boiling points.But this extra procedure tends to induce additional error and it is laborious and time-consuming.A derivatization-free method was therefore established in the present investigation to determine FFAs in human plasma by capillary（GC-FID）.After extraction of FFAs from plasma,a highly polar FFAP（free fatty acid in plasma） column was employed to directly quantitate FFAs concentration,free from derivatization reaction.All sample pretreatments were carried out at room temperature,improving recovery of short-chain FFAs.Heptadecanoic acid（C17：0） was employed as internal standard,and the proposed method was validated for recovery,precision,sensitivity,stabi-lity,and linearity.Validation data show that it is suitable for clinical study that has been applied to the evaluation of FFAs levels in plasma of diabetic nephropathy（DN） patients during the course of treatment.Forty-seven patients diagnosed with DN were admitted to the double-blind experiment.Control group（n=17） underwent solely basic treatment and the patients did not show significant change in FFAs concentration during six months of treatment.Experiment group（n=30） was supplied with traditional Chinese medicine besides basic treatment.After six months of medication,their plasma concentration of palmitic acid（C16：0）,stearic acid（C18：0） and oleic acid（C18：1n-9） decreased while linolenic acid（C18：3n-3） increased significantly（P〈0.05）.These four compounds could be served as biomar-kers in the evaluation of drug efficacy,and their quantitation in plasma may provide additional information for disease progression in DN patients.
基金supported by the Foundation of Shaanxi Provincial Science and Technology Plan Projects (2007K14-04)
文摘Objective To investigate the therapeutic effect of intensive glycemic control on patients with early diabetic nephropathy. Methods A total of 41 type 2 diabetes patients who developed microalbuminuria were divided into two groups randomly. Patients in Group A received intensive glycemic control and the blood glucose in Group B was regularly controlled. Glycemic monitoring and control were followed for 12 weeks to observe the changes of microalbuminuria in both groups; meanwhile the levels of serum lipids and coagulation indices were also recorded. Results The urine albumin excretion rate (UAER) in Group A decreased significantly from (47.91±13.86)mg/24h to (35.31±14.56)mg/24h after 12 weeks (P<0.05),and this decrease was significantly greater than that in Group B. However,Group B had no significant difference in UAER decrease [(48.93±13.32)mg/24h to (40.48±19.62)mg/24h,P>0.05]. The decrease of triglyceride (TG) and low-density lipoprotein cholesterol (LDL cholesterol),and the increase of high-density lipoprotein cholesterol (HDL cholesterol) showed no significant differences (P>0.05). And the level of plasma fibrinogen (FIB) showed no significant decrease after 12 weeks,either (P>0.05). Conclusion Intensive glycemic control reduces the level of microalbuminuria and may ameliorate the progression of early diabetic nephropathy.
文摘Background: Diabetes mellitus (DM) is the leading cause of end stage renal disease (ESRD) worldwide. Although DM with proteinuria is the ultimate result of diabetic nephropathy (DN), a wide spectrum of non-diabetic renal diseases (NDRD) can occur in such patients. Objective: To observe the frequency and histological pattern of NDRD in diabetic patients with proteinuria and to explore their association with clinical and laboratory parameters. Methods: This cross-sectional study was conducted in the Department of Nephrology, Bangabandhu Sheikh Mujib Medical University (BSMMU), Dhaka, Bangladesh from April 2016 to September 2017. In this study a total of 38 cases of DM with proteinuria (>1 gm/24-hour) were selected purposively. Renal biopsy was done in all patients. Based on histological findings they were categorized into two groups;Group 1 with NDRD and Group II with DN. Their clinical and laboratory parameters were analyzed and compared. Results: Among the total study subjects, 21 (55.3%) were male and 17 (44.7%) were female, mean (±SD) age 43.45 ± 9.99 years in the NDRD group and 41.57 ± 9.50 years in the DN group. Thirty one cases (81.6%) out of thirty eight had NDRD and seven (18.4%) cases had isolated DN;therefore more than two third cases had NDRD. Duration of DM was found to be significantly shorter (p = 0.004) in the NDRD group. Diabetic retinopathy was present in 12.9% cases in NDRD group vs. 57.1% cases in DN group (p = 0.025). Frequency of microscopic hematuria was significantly higher (90.3%) in NDRD patients (p = 0.002). Conclusion: The frequency of NDRD in type 2 diabetic patients other than diabetic nephropathy is relatively high. Membrano proliferative glomeru-lonephritis and membranous nephropathy are more common in NDRD. Absence of diabetic retinopathy, presence of hematuria and shorter duration of DM are markers associated with NDRD in type 2 DM, which are important indicators for renal biopsy in diabetic patients with proteinuria.
文摘【目的】观察苦瓜皂苷G对糖尿病肾病(DN)大鼠的治疗作用及机制。【方法】从55只大鼠中随机抽取45只采用一次性腹腔注射链脲佐菌素(STZ)法诱导DN模型。将造模成功的38只大鼠随机分为模型组9只、苦瓜皂苷G低剂量组9只、苦瓜皂苷G中剂量组10只、苦瓜皂苷G高剂量组10只。剩余10只大鼠设为正常组。对应灌胃给药4周后,酶联免疫吸附分析(ELISA)检测大鼠24 h尿蛋白水平,血糖仪检测空腹血糖(FBG),全自动生化分析仪检测血肌酐(SCr)、血尿素氮(BUN)水平,苏木素-伊红染色(HE)法观察大鼠肾组织病理学变化,采用实时定量聚合酶链反应(RT-qPCR)和Western Blot法分别检测E-钙黏蛋白(E-cad)、α-平滑肌肌动蛋白(α-SMA)、Notch1、锌指蛋白转录因子1(Snail1)、Jagged1 m RNA及蛋白表达情况。【结果】与正常组比较,模型组24 h尿蛋白、FBG、血清SCr和BUN水平及肾组织α-SMA、Notch1、Snail1、Jagged1 mRNA及蛋白水平显著升高,肾组织E-cad m RNA及蛋白水平降低,肾组织可见肾小球萎缩、肾小管扩张及间质纤维化;与模型组比较,苦瓜皂苷G低、中、高剂量组24 h尿蛋白、FBG、血清SCr和BUN水平及肾组织α-SMA、Notch1、Snail1、Jagged1 mRNA及蛋白水平显著降低,肾组织E-cad m RNA及蛋白水平升高,肾组织病理损伤得到改善。【结论】苦瓜皂苷G可有效改善DN大鼠肾功能障碍、减轻肾脏纤维化,其机制可能与通过调控Notch/Snail1信号通路,抑制α-SMA和Jagged1 mRNA及蛋白表达,增强E-cad mRNA及蛋白表达有关。
