In this paper, absorption and pharmacokinetic study of Radix Rehmanniae was studied by liquid chromatography coupled with mass spectrometry method after oral administration to rats. By comparing the chromatograms of ultraviolet, full scan, extracted ion and selective reaction monitoring (SRM) of standard solution, Radix Rehmanniae, blank plasma and rat plasma post drug administration, catalpol and ajugol were found to be the main compounds absorbed from Radix Rehmanniae. Plasma concentrations of aucubin, dihydrocatalpol, rehmannioside A (or rehmannioside B/ melittoside) and rehmannioside D were very low. Quantitative method for catalpol and aucubin and semi-quantitative method for other compounds in rat plasma were established. The pharmacokinetic study of those absorbed components was conducted after oral administration of 6 g x kg(-1) Radix Rehmanniae water extract to rats. Cmax, t(1/2) and AUC(0-infinity) of catalpol and ajugol were (2349.05 +/- 1438.34) and (104.25 +/- 82.05) ng x mL(-1), (0.86 +/- 0.32) and (0.96 +/- 0.37) h, (4407.58 +/- 2734.89) and (226.66 +/- 188.38) ng x h x mL(-1), respectively. tmax was at 1.00 h for catalpol and ajugol. Both catalpol and ajugol were absorbed and excreted rapidly.

The objective of study was to investigate the effect of liposomal transfection of antisense oligodeoxynucleotide (ASON) on alpha-globin gene expression and proliferation of K562 cells, to explore the new way of gene therapy in beta-thalassemia. Targeted ASON of alpha-globin was designed and synthesized, and compared with positive control [sense oligodeoxynucleotide (SON) group] and blank control. By liposomal transfection, ASON, SON was co-cultured with K562. The efficiency of transfection was assayed by fluorescence microscopy and flow cytometry (FCM), the alpha-globin gene expression of K562 was measured by real-time PCR, and the proliferation of K562 was determined by Cell Count Kit-8 assay. The results indicated that the highest efficiency was at 24 hours after liposomal transfection, the gene expression level of alpha-globin in ASON group was significantly lower than that in SON group and blank control (p < 0.01). The proliferation of K562 cells was obviously inhibited, meanwhile the above effect showed the dose-dependent manner. It is concluded that the liposomal transfection of ASON inhibits the alpha-globin gene expression of K562 cells, which may be the new target for gene therapy in beta-thalassemia.
Cell Proliferation ; drug effects ; Gene Expression ; Humans ; K562 Cells ; Liposomes ; metabolism ; Oligonucleotides, Antisense ; pharmacology ; Transfection ; alpha-Globins ; metabolism

Objective To compare the clinical outcomes of gonadotropin-releasing hormone (GnRH) antagonist (GnRH-ant) fixed protocol with GnRH agonist (GnRH-a) long protocol in infertile patients with normal ovarian reserve function in their first in vitro fertilization-embryo transfer (IVF-ET) cycle,and to explore the feasibility and advantage of GnRH antagonist protocol performed in normal responders.MethodsFrom January 2011 to June 2011,771 infertile women with normal ovarian reserve function underwent their first IVF or intracytoplasmic sperm injection (ICSI) cycles in Peking University Third Hospital,which were divided into 245 cycles in GnRH-ant fixed protocol group ( GnRH-ant group) and 526 cycles in GnRH-a long protocol group ( GnRH-a group).The data of general demographic,treatment and clinical outcome were compared between two groups.ResultsAge,infertile duration,body mass index (BMI),baseline serum follicle-stimulating hormone (FSH) and estradiol levels between two groups did not reached statistical difference (P ＞ 0.05 ).The level of estradiol was (12 289 ± 6856) pmol/L in GnRH-ant group and (14934±8007)pmol/L in GnRH-a group at day of hCG injection.The mean length of stimulation was ( 10.3 ± 1.2) days in GnRH-ant group and ( 12.8 ± 1.6) days in GnRH-a group.The dose of gonadotropin was (2013 ± 607 ) U in GnRH-ant group and (2646 ± 913 ) U in GnRH-a group.The number of ovum was 15 ± 7 in GnRH-ant group and 17 ± 8 in GnRh-a group.Those clinical parameter all reached statistical difference (P ＜0.05 ).The number of embryo was 7 ±4 in GnRH-ant group and 8 ± 5 in GnRH-a group,the rate of clinical pregnancy was 40.9％ (94/230) in GnRH-ant group and 45.6％ (216/474)in GnRH-a group,the rate of implantation was 26.1％ (128/490)in GnRH-ant group and 30.9％ (307/994) in GnRH-a group,the rate of continuing pregnancy was 38.7％ ( 89/230 ) in GnRH-ant group and 42.6％ (202/474) in GnRH-a group,those parameter did not reach statistical difference (P ＞ 0.05).The rate of moderate or severe ovarian hyperstimulation syndrome was 2.4％ ( 6/245 ) in GnRH-ant group and 4.2％ (22/526) in GnRH-a group,which did not show significant difference ( P ＞ 0.05 ).ConclusionIn the first IVF or ICSI cycle of the patients with normal ovarian reserve function,the fixed GnRH-ant protocol could get the same satisfied clinical outcome,and it is more economic,convenient and safer compared with low dose depot GnRH-a long protocol.

Objective To describe and compare the roles of 3 external quality assessment programs in assessment of analytical quality of serum creatinine and urea.Methods Research in quality management methods.Sixty-five laboratories those enrolled in the Natonal Center for Clinical Laboratories′programs of routine chemistry external quality assessment ( EQA ) , trueness verification ( TV ) for small molecular metabolites and external comparison of internal quality control ( IQC) simultaneously in 2013 were selected , the performances of those laboratories of serum creatinine (crea) and urea in terms of total errors(TE), bias and CV were obtained by using the above 3 programs, and these performance were assessed against the criterion listed in the analytical quality specifications for routine analysis in clinical biochemistry ( WS/T 403-2012).The failure ratio of 65 laboratories on each performance was calculated , the sensitivity of 3 external quality assessment programs in detection of analytical quality deficiency among clinical laboratories were compared.Results Only 1 laboratory failed in the 1st routine chemistry EQA in terms of TE of creatinine , failure ratio is 1.5%(1/65).Three laboratories failed in the 2nd EQA and caused a failure ratio of 4.6%(3/65).For serum urea, 3 laboratories failed in the 1st routine chemistry EQA with a failure ratio of 4.6%(3/65).Two laboratories failed in the 2nd EQA with a failure ratio of 3.1%(2/65).The failure ratios of creatinine determination in two samples in TV were 41.5%(24/65) and 21.5%(14/65) respectively, and the failure ratio of urea determination were 53.8%( 36/65 ) and 32.3%( 21/65 ) respectively.In the program of external comparison of IQC , the CVs of creatinine and urea determination ranged from 0.7% to 6.2%and from 1.0%to 7.2%respectively, their respective failure ratio range were 15.4%(10/65) and 40.0%(26/65).The failure ratio in routine EQA were much less than those in the other two programs , the laboratories failed in routine EQA program were all failed in trueness verification or /and the comparison of IQC programs, but not vice versa.Conclusions By participating in the programs of routine EQA , TV and comparison of IQC laboratories could assess the performances of inaccuracy , bias and imprecision.Laboratories should participate in different external quality assurance programs to detect their quality issues and get improved.

Objective To analyze the clinical outcomes of patients treated with in vitro fertilization and embryo transfer (IVF-ET) and influence factors of pregnancy rate. Methods We retrospectively analyzed the clinical data of 12 491 cycles, including 6832 fresh IVF/intracytoplast single sperm injection (ICSI) cycles and 5659 frozen embryo transfer (FET) cycles from 2005 to 2007. Results The clinical pregnancy rate per cycle was 32. 99% (2254/6832)in fresh embryo transfer program, and the live birth rate was 25.75% (1394/5413); the early pregnant loss rate was 9. 36% (211/2254), and the prenatal defect rate was 1.45% (25/1722). Through analysis of these patients' basic data, we found that the patients' age, causes for infertility, egg retrieval and cycle number affected the pregnancy rate. Using logistic regression method, we found that patients′ age was the most important factor affecting pregnancy outcome. In FET cycles, the clinical pregnancy rate was 38.08% (2155/5659), significantly higher than fresh embryo transfer cycles. Conclusions IVF-ET treatment is a safe and effective method for infertility couples. However, the female age and poor ovarian response are the main factors affecting pregnancy rate. Thawed embryo transfer can increase the accumulated pregnancy rate effectively.

External quality assessments play important roles in quality improvement in clinical laboratories, but most laboratories focused on the unsatisfied data only.With appropriate quality controls, laboratories can detect not only the error sourses of unsatisfied data but the potential error sourse of satisfied data as well.

Objective To evaluate the commutability of certified reference material IRMM ERM-DA 471/IFCC for cystatin C measurement among 8 methods.Methods 46 individual samples used for the commutability study were residual serum samples collected from clinical laboratories.The individual samples interspersedwith pooled sera and the certified reference material IRMM ERM-DA 471/IFCC and the whole set of samples was divided into 2 subsets for measurements in 2 days.The measurements were performed by8 different methods o.The samples were measured in duplicate order.Calibration was performed every day. Passing-Bablok regression was performed to compare the slopes and intercepts of mean values of the serum samples derived from different methods. Pearson correlation cofficient was also calculated. Deming regeression and 95%confidence intervals were calculated to evaluate the statistics commutability of ERM-DA 471/IFCC.The minimal specification of bias derived from biological variations was calculated to evaluate the clinical commutability.Results The within-laboratory CVs of pool sera ranged from 0.5% to 4.0%.The Passing-Bablok slope ranged from 0.765 to 1.311 and intercepts ranged from -0.04 to 0.241.The determination coefficient of Pearson regression ranged from 0.988 to 0.999.Deming regeression and 95%confidence intervals demonstrated commutability of ERM-DA 471/IFCC in 4/28 (14.3%) methods pairs. The minimal specfication bias ( 5.12%) demonstrated commutability of ERM-DA 471/IFCC in15/28 (53.6%) methods pairs.Conclusions The ERM-DA 471/IFCC domonstrated poor commutability between some methods pairs. The commutability of ERM-DA 471/IFCC should be evaluated before used as calibrators.

Objective To study the applicability of a new analytical specification defined in WS/T 403-2012 in the external quality assessment schemes and the external comparision of internal quality control .Methods It was a quality management method study.Total error allowable criterions listed in WS/T 403-2012 and GB/T 20470-2006 were selected to assess the results of 23 analytes in the 1st challenge of 2013 routine chemistry external quality assessment.The acceptable rate of 23 analytes were calculated with the two specifications.Criterions of imprecision derived from the two standards were applied to assess the coefficient of variation with internal quality control data.Results With the specification based on WS/T 403-2012, the ratio of laboratories that all five samples were passed in the 1st challenge for 23 analytes ranged from 55.5%to 94.7%.The ratio of laboratories with 80%or more samples passed in 2013 EQA ranged from 73.9%to 98.5%.While ratios of two kinds described above evaluated based on GB /T 20470-2006 ranged from 63.0%to 99.2%, and from 90.0% to 99.7%, respectively.The acceptable rate of CV according to the two criterions ranged from 55.5% to 94.7% and 63.0% to 99.2%, respectively.Conclusions As evaluation criterions of external quality assessment allowable total error and internal quality control imprecision in routine chemistry , the specification in WS/T 403-2012 can be used to assess the analytical performance of clinical laboratory more objectively and comprehensively.It can help laboratories to identify the latent problems for further quality improvement.

Objective To investigate the relationship between the imbalance of suppressors of cytokine signaling 1(SOCS1)/SOCS3 and abnormal activation of monocytes/ macrophages in children with acute-phase asthma, and explore the molecular mechanism of chronic inflammatory process on airway.Methods The present study enrolled 20 asthmatic children and 20 age-matched normal children. Dual-color flow cytometric analysis was performal to detect the percentage of CD_ 80 、CD_ 86 expressing on CD_ 14 ~+ cell. Reverse transcriptase-polymerase chain reaction and real-time PCR were used to analyze SOCS1,SOCS3 expression in monocytes/ macrophages.Results The proportions of CD_ 80 ,CD_ 86 expressed on CD_ 14 ~+ cell in children with asthma were significantly higher than those in control subjects(CD_ 80 :7.0% vs 1.70%),(CD_ 86 :11.37% vs 2.03%),all P

Constructing prokaryotic expression vector pKY-RMBAY by gene recombination and research its optimizing productive conditions.By PCR technology synthesizing the gene of the RMBAY with preference codon of E.coli and the RMBAY gene was inserted into high efficiency expression vector pKYB-MCS.Expressed fusion proteins in E.coli ER2566 were purified with Chitin-Beads column.Fusion proteins binding on Chitin-Beads was cut on N-terminus of intein due to the induction of ?-mercaptoethanol and the target peptide RMBAY was released.The RMBAY was identified by mass spectrum.Experiment results showed RMBAY can be high efficiently expressed in E.coli ER2566,with optimizing productive conditions the yield of the RMBAY may be 6.7mg/L fermentation product and its purity is greater than 98%.The molecular weight of RMBAY is 3.887 kDa by mass spectrum and that accords with its theory value.