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【Objective】 To study the serological characteristics and genetic background of 20 samples with ABO blood group discrepancies in Shenyang. 【Methods】 Serological test, polymerase chain reaction-sequence specific primer (PCR-SSP) and sequencing of the full coding of ABO gene and the Intron 1 were conducted in 20 samples with ABO blood group discrepancies. 【Results】 Ten subtypes (Am, Bw, Bx, B3,, A2B, AxB, A2Bw, A2Bx, AwB and ABw) were detected in 20 samples, with AB subtype as the dominant. Sixteen ABO alleles were found, including 5 common alleles (A1.01, A1.02, B. 01, O. 01.01 and O. 01.02), nine rare alleles (AW.37, BW.03, BW.08, B3.07, cisAB.02, cisAB.03, cisAB.06, BA.04 and O. 01.04) and two novel alleles (AM.03 and cisAB.07). The AM.03 allele had a nucleotide change at position 912 (C to A) compared with A1.02 allele, which resulted in an amino acid substitution (S304R). The cisAB.07 allele was observed a missense mutation at position 797 (T to C) which resulted in an amino acid substitution (M266R) compared with B. 01 allele. The serologic had been changed, and both A antigen and B antigen were expressed. 【Conclusion】 The study revealed the genetic background of 20 samples with ABO blood group discrepancies, and two new alleles as ABO*AM.03 (912 C to A) and ABO*cisAB.07 (797 T to C) were first reported.
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【Objective】 To study the molecular basis of D variant and explore the molecular genetic mechanism of novel weak D alleles. 【Methods】 Blood samples were screened for D variants by serological method. The nucleotide sequences of coding region were amplified by PCR and sequenced directly, and RHD gene heterozygosity was detected. 【Results】 Weak D phenotype was confirmed by serological test, and two novel alleles were detected by DNA sequencing. The first was novel weak D 1102A allele, 1102G>A mutation in exon 8, resulting in a 368Glu>Arg substitution in two samples. The second was novel weak D 399C allele, carried a 399G>C mutation in exon 3, which led to a 133Lys>Asn substitution. 【Conclusion】 In this study, D variants were detected by sequence-based typing, and two new alleles were identified.
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Objective: To screen the effective hemostasis fraction of Balanophora involucrata. Methods: Balanophora involucrata was extracted by 95% ethanol using reflux extraction, and the residue was extracted by water. The filtrate was combined and concen-trated, and then extracted by petroleum ether, ethylacetate and n-butylalcohol in turn to respectively obtain petroleum ether phase, eth-ylacetate phase, n-butylalcohol phase and H2O phase. The effective hemostasis fraction was determined by measuring the time of blood coagulation and hemorrhage combined with plasma recalcification time. Results: Compared with the blank group, ethylacetate extract from Balanophora involucrata could significantly shorten the time of blood coagulation and hemorrhage, and the plasma recalcification time (P<0. 01). n-Butylalcohol group at high dose could shorten the time of hemorrhage and the plasma recalcification time (P<0.05),while had no effect on the time of blood coagulation(P>0.05). Petroleum ether extract and H2O phase couldn't shorten the time of blood coagulation and hemorrhage, and the plasma recalcification time(P>0. 05). Conclusion: The effective hemostasis frac-tion of Balanophora involucrata mainly concentrates in ethylacetate extract.
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Objective To explore the effect of total saponins from Medicago polymorpha (TSMP) on glucose metabolism in type 2 diabetes mellitus (T2DM) rats and its possible mechanism.Methods T2DM rats were established by feeding with fat diets and intraperitoneally injecting with STZ 30 mg/kg.The rats were divided into control group,model group,metformin group (0.2 g/kg),TSMP high dose group (1.4 g/kg) and TSMP low dose group (0.7 g/kg),which were administrated for four weeks.At the end of administration,blood samples were collected to determine insulin resistance index (IRI),levels of fasting blood glucose (FBG),glycosylated hemoglobin (HbAlc),hepatic glycogen,interleukin (IL)-1 β,tumor necrosis factor (TNF)-α,free fatty acids (FFA) and Leptin,and activities of pyruvate kinase (PK),hexokinase (HK),glucose-6-phosphatase (G-6-Pase),fructose-1,6-bisphosphatase and glucokinase (GK).Western blot was used to detect the expression levels of G-6-Pase and phosphoenolpyruvate carboxykinase (PEPCK) proteins.Results Compared withthe model group,high dose (1.4 g/kg) TSMP decreased IRI,levels of FBG,HbA1 c,IL-1β,TNF-α,FFA and Leptin,and activities of G-6-Pase and fructose-1,6-bisphosphatase in T2DM rats (P＜ 0.05),while increased level of hepatic glycogen,and activities of PK,HK and GK (P＜0.05).Moreover,high dose (1.4 g/kg) TSMP down-regulated expression levels of G-6-Pase and PEPCK protein in liver tissues.Conclusion Chronic administration of TSMP can improved glucose homeostasis in T2DM rats,which might be related to promoting utilization of glucose,and alleviating inflammatory and insulin resistance.
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Objective To explore the effect of total saponins from Medicago polymorpha (TSMP) on glucose metabolism in type 2 diabetes mellitus (T2DM) rats and its possible mechanism.Methods T2DM rats were established by feeding with fat diets and intraperitoneally injecting with STZ 30 mg/kg.The rats were divided into control group,model group,metformin group (0.2 g/kg),TSMP high dose group (1.4 g/kg) and TSMP low dose group (0.7 g/kg),which were administrated for four weeks.At the end of administration,blood samples were collected to determine insulin resistance index (IRI),levels of fasting blood glucose (FBG),glycosylated hemoglobin (HbAlc),hepatic glycogen,interleukin (IL)-1 β,tumor necrosis factor (TNF)-α,free fatty acids (FFA) and Leptin,and activities of pyruvate kinase (PK),hexokinase (HK),glucose-6-phosphatase (G-6-Pase),fructose-1,6-bisphosphatase and glucokinase (GK).Western blot was used to detect the expression levels of G-6-Pase and phosphoenolpyruvate carboxykinase (PEPCK) proteins.Results Compared withthe model group,high dose (1.4 g/kg) TSMP decreased IRI,levels of FBG,HbA1 c,IL-1β,TNF-α,FFA and Leptin,and activities of G-6-Pase and fructose-1,6-bisphosphatase in T2DM rats (P＜ 0.05),while increased level of hepatic glycogen,and activities of PK,HK and GK (P＜0.05).Moreover,high dose (1.4 g/kg) TSMP down-regulated expression levels of G-6-Pase and PEPCK protein in liver tissues.Conclusion Chronic administration of TSMP can improved glucose homeostasis in T2DM rats,which might be related to promoting utilization of glucose,and alleviating inflammatory and insulin resistance.
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Objective To establish fingerprint and mutual mode of aqueous extracts of Eucommia ulmoides Oliv.from different regions by high performance liquid chromatography ( HPLC). Methods The column of SinoChrom ODS￣BP (250 mm×4.6 mm, 5 μm) was used.The mobile phase consisted of acetonitrile￣0.1% phosphoric acid with gradient elution.The flow rate was 1.0 mL.min-1 , the detective wavelength was 230 nm, and the column temperature was 25 ℃ . Results The fingerprint consisted of 14 common peaks.The overall similarity for ten batches of Eucommia ulmoides Oliv.from different regions was 0.592-0.986.The standard fingerprint of Eucommia ulmoides Oliv.was established by HPLC. Conclusion This fingerprint method is simple and repeatable, and it provides a scientific basis for the quality control of Eucommia ulmoides Oliv.