Objective To investigate the changes in histological characteristics and collagen content in uterosacral and cardinal ligaments of pefimenopausal women in relation to relaxation of pelvic supports.Methods Twenty-eight subjects undergoing hysterectomies were selected,in which 14 cases were perimenopausal women with relaxation of pelvic support as the relaxation group and 14 women at perimenopansal age with leomyoma,cervical cancer,adenomyosis as the control group.Samples of cardinal ligaments and uterosacral ligaments were obtained at hysterectomies,and the tissues were shced and stained bv Masson's trichrome technique.Histological characteristics of the samples were studied and immunohistochemistry assay was applied to demonstrate the contents of collagen types I andⅢ.Results (1)The collagen in uterosacral ligaments and cardinal ligaments were stained blue by tlle MaSson's trichome technique.In comparison to the control group,the relaxation group had milder positive stains of the collagen and the stains were distributed in unequal intensities.Collagen content was arranged in loose pattern.Focal arrangement of the collagen was dense but fragmented.Collagen fibers were atrophic. (2)In immunohistochemistry assay and image analysis,collagen was positive in light to deep brown areas.In the relaxation group,positive units of collagen types I and III in cardinal ligaments were 13.8±2.1 and 9.6±2.4 respectively.Positive units of coHagen types I and Ⅲ of cardinal ligaments in the control group were 27.4 ±3.5 and 17.7 ±4.0 respectively.Difierences between these two groups were statistically significant (P

Objective To investigate current status and problems of coagulation factor Ⅷ and Ⅸ assay in domestic laboratories so as to provide the reference for implementing the standardization and quality improvement.Methods A questionnaire survey was carried out in 76 laboratories,and quality control materials were distributed to 54 laboratories for activity assay.The questionnaire information was analyzed statistically.Test results of quality control materials were classified into three groups according to the reagents and the ranked grading analysis were used to evaluate the performance.Results This research was investigative study.The amount of sample was less than 30 per month in 72％ (52/72)of laboratories.The frequencies of calibration were different,and 33％ (24/72) of laboratories did not perform calibration in a different assay batch.39％ (28/72)of laboratories did not run internal quality control,and about 21％ (15/ 72) of laboratories just performed the normal level quality control.Individual laboratories showed a high cumulative CV (＞ 30％) of intemal quality control.For normal FⅧ and FⅨ control materials,the CV of results were 11.3％-18.2％ and 11.3％-17.9％ respectively as well as 15.3％-20.3％ and 19.5％-21％ for abnormal.Of the three groups,the proportions of laboratories which the FⅧ test results out with consensus were18％,24％ and 22％ as well as 20％,24％ and 28％ for FⅨ.Conclusions The key requirements for quality control of coagulation factors active assay remain to be addressed and implemented.The repeatability and comparability in some laboratories are not satisfactory to meet the clinical needs.With the purpose of promoting quality improvement,we need to develop guidelines,organize related training and establish a national external quality assessment scheme.

Objective To study the single nucleotide polymorphisms (SNP) in 3 sites allele (T189M, R85H, C121W) of SCN1B and the association between gene distribution and epilepsy. Methods All 330 blood samples of refractory (80 cases), non-refractory (100 cases) epilepsy patients and healthy people (150 cases) were collected. Genomic DNA of leucocyte was extracted. SNPs of three sites allele of SCN1B were tested by allele-specific primer-polymerase chain reaction (ASP-PCR).Data were analyzed by SAS 8.1 statistical software. Results Epilepsy group and healthy group had significantly statistical difference in composition of 3 sites allele on single site genotype (x~2=11.19, 11.14 and 6.50, all P < 0.05).There was no statistical significance between refractory and non-refractory epilepsy group. On gene combination, in 27 different combinations of polymorphism, mutation frequency in 3 sites (CT + AG + CG) was highest in epilepsy group (18.40%).The next was one site in CT + GG + CC (16.80%).In healthy group, frequency of non-variant in CC + GG + CC was highest (16.67%), and the next was 2 sites in CT+ AG+CC (13.73%).Thirty-five cases in epilepsy group (28.80%) had 3 sites mutation compared with 10 cases in healthy group (9.71%), and their difference had statistical significance (x~2=12.54, P<0.05).Eighteen cases in refractory epilepsy group (30.51%) had 3 sites mutation compared with 21 cases in non-refractory epilepsy group (28.77%), and the difference had no statistical significance. Fifty cases in epilepsy group (40.00%) had 2 sites mutation compared with 41 cases in healthy group (40.20%), and there was no statistical significance between them; 25 cases in refractory epilepsy group (42.37%) had 2 sites mutation compared with 21 cases in non-refractory epilepsy group (28.71%), and their difference had no statistical significance. Conclusions Mutation, especially multisite mutation of SCN1B is relatively likely to cause epilepsy in human. Gene distribution and combination of three sites allele of SCN1B in refractory epilepsy is close to that in non-refractory epilepsy.

ObjectiveTo establish an effective method for F9 gene mutation detection by using high resolution melting ( HRM ) curve analysis.Methods Peripheral blood samples of 55 hemophilia B (HB) patients were collected from Shanghai Ruijin Hospital during January 2005 to June 2010.Genomic DNA was extracted from the peripheral blood.PCR amplification combined with sequencing was used to identify the F9 gene mutations in 40 patients.HRM assay was established on the 21 DNA samples with known mutations in exonl to exon7 of F9 gene.Mutation scanning of exonl to exon7 by HRM combined with direct sequencing of exon8 was used in the molecular diagnosis of 15 HB patients with unknown F9 gene mutations.ResultsF9 gene mutation was detected in each of the 40 HB patients by direct sequencing.By HRM,the different melting curve patterns were identified in 19 out of 21 cases.The detection rate was about 90％.Through mutation scanning of exonl to exon7 by HRM combined with direct sequencing of exon8,F9 gene mutations were detected in all the 15 HB patients with unknown F9 gene mutations.Thirty-four F9 gene mutations had been identified in the 55 HB patients.ConclusionsA new strategy of HB genetic diagnosis,scanning mutations of exonl to exon7 combined with DNA sequencing of exon8 of F9 gene,is established in this study.The new strategy is efficient and reliable.

Objective To study two new factor Ⅸ mutations Cys82Ser and Ile288Ser in vitro and research the molecular mechanism of haemophilia B. Methods PcDNA3. 1 ( - ) FⅨwt expression plasmid was prepared. The mutated FⅨcDNA expression plasmids, PcDNA3.1 ( - ) FⅨM1 (Cys82Ser) and PcDNA3. 1 ( - ) F Ⅸ M2 (Ile288Ser) were constructed by megaprimer method respectively. Transient expression experiments were performed using HEK293 cells transfected with the expression vectors containing the wild-type or the mutation recombinant cDNA. PcDNA3. 1 ( - ) was used as a blank control. The expression proteins were detected by ELISA, factor activity assay and flourescence stain. Results The results suggested that the two FⅨ gene mutations did not induce the reduction of the mutant FⅨ mRNA compared with the wild-type FⅨ mRNA. The FⅨ:Ag in culture media and cell lysate of wild type conduct were assigned as 100. 0%. The results of PcDNA3.1 ( - ) FⅨ M1 mutation protein were (27. 1 ± 5. 2)% and (99.4 ±4. 1)% respectively. For PcDNA3. 1( - )FⅨM2, the results were (5.3 ± 1.8)% and (31.7 ±2. 5)% respectively. The FⅨ: C in culture media of wild type conduct was also assigned as 100. 0%. Then the two types of mutant protein were ( 8. 5 ± 3.2 ) % and ＜ 1%, respectively. Immunofluorescence microscopy result suggested that the intensity of perinuclear spot was reduced in cells transfected with PcDNA3.1 ( - ) FⅨM2 while staining for PcDNA3. 1 ( - ) FⅨM1 was predominantely diffuse without perinnclear enhancement. Conclusions These results strongly suggest that the FⅨ Cys82Ser mutation protein is not been correctly folded, by any possibility. The mutation protein has secretion defect. The secretion dysfunction and the protein degradation intracellular are possiblely the molecular pathology of Ile288Ser mutant protein.

BACKGROUND:Many studies have demonstrated that FirebirdTM and Taxus Express2TM can effectively reduce intrastent restenosis.However,there are few data about long-term efficacy of two stents,and reports about middle and long-term follow up are rare.OBJECTIVE:To observe the safety and biocompatibility of FirebirdTM and Taxus Express2TM in percutaneous coronary intervention(PCI) for coronary artery disease.DESIGN,TIME AND SETTING:Non-randomized concurrent control clinical observation was performed at Department of Cardiology,Hongqi Hospital of Mudanjiang Medical College from April 2005 to April 2008.PARTICIPANTS:233 patients(268 lesions) undergoing PCI were divided into FirebirdTM(n =82),Taxus Express2TM group(n =80) and bare metal stent group(n =71).METHODS:Coronary arteriography was performed through femoral artery or radial artery.Vascular inner diameter was determined using quantitative computer analysis.The patients underwent FirebirdTM,Taxus Express2TM and bare metal stenting,respectively.The patients were reexamined and followed-up using telephone every 2-4 weeks after discharging and examined using coronary arteriongraphy after 9-12 months.The follow-up lasted for 24 months.MAIN OUTCOME MEASURES:Characteristics of arteriongraphy and stenting condition of all patients;biocompatibility of stent to host;major adverse cardiac events during hospitalization and follow-up,including death,angina pectoris attacks or heart failure;coronary artery diameter decreased ≥ 50% was regarded as restenosis.RESULTS:101 stents were implanted in Firebird group,98 in Taxus Express2 group and 85 in bare metal stent group.There was no stent defluxion,dislocation,or breakage.No noticeable platelet decrease,hemolysis or white blood cell increase was found.There were no significant differences among three groups in terms of Characteristics of arteriongraphy and stenting condition.The incidence of major adverse cardiac events and intravascular restenosis in Firebird and Taxus Express2 groups was fewer than bare metal stent group(P 0.05).CONCLUSION:No specific biocompatibility responses in treatment of coronary artery diseases using FirebirdTM and Taxus Express2TM.The two drug-eluting stents are superior over bare metal stent in reducing restenosis.The safety and efficacy of two drug-eluting stents are similar.

Objective To construct the eukaryotic expression vector pEGFP-N1/IL-37b and analyze the expression of IL-37 gene in THP-1 cells. Methods Total RNA was extracted from human peripheral blood mononuclear cells ( PB-MCs) and the coding region of IL-37b gene was amplified by RT-qPCR. Then, the gene was cloned into pEGFP-N1 eu-karyotic expression vector. After transfected the recombinant plasmid into THP-1 cells, the expression of IL-37 was detec-ted by RT-qPCR and Western blot. Results Double restriction enzyme digestion and gene sequencing showed that IL-37b gene was correctly inserted into the eukaryotic expression vector pEGFP-N1. RT-qPCR and Western blot showed that the IL-37 expression level was increased significantly (P<0. 01) after transfection in THP-1 cells. Conclusions We successful-ly constructed a novel anti-inflammatory cytokine IL-37 eukaryotic expression vector pEGFP-N1/IL-37b, which lays a foun-dation for further study on IL-37 functions and its association with related diseases.

Objective To evaluate the promotional effectiveness of different TCM appropriate technology packages; To provide a scientific basis for improving the technology package. Methods 8 assessment indexes, including input costs, the number of experts, the number of people with senior professional post, the number of grassroots people under the guidance of experts, the number of training hours, the number of people receiving training, evaluating rate and the number of users were selected. TOPSIS method was used to evaluate the promotional effectiveness of five kinds of TCM packages in grassroots. Results The Ci values for appropriate technology packages were 0.76 for pains in neck, shoulder, waist and lower extremities, 0.49 for gynecological diseases, 0.44 for infantile diarrhea, 0.66 for chronic gastropathy, and 0.00 for tumors. Conclusion The effectiveness of promotion of TCM appropriate technology packages from high to low is pains in neck, shoulder, waist and lower extremities package, chronic gastropathy package, gynecological diseases package, infantile diarrhea package, tumors package.

OBJECTIVE:To provide empirical reference for protecting the supply of clinical drugs. METHODS:Literature re-view was combined with the conditions of drugs that lost the bidding in the centralized bidding in Anhui province to determine the short drugs need to investigate. Questionnaire was adopted to investigate the situations and reasons of short drugs in 5 third-grade class-A hospitals of 5 areas in Anhui province from Nov. 2013 to Oct. 2014. RESULTS:A total of 5 questionnaires were sent out, and 5 were received with effective response rate of 100%. There were totally 54 short drugs in the 5 third-grade class-A hospitals, including the most serious shortage of drugs for neurocirculatory system,accounting for 20.37%. Shortage was mainly due to the low price of drugs,accounting for 48.15%,and insufficient supply,less suffering patients/low dosage and other reasons. CON-CLUSIONS:In view of the shortage of drugs,government departments should improve the drug pricing and bidding policy,pro-duction enterprises should enhance the enterprise production and development capabilities,business companies should optimize the distribution pattern of network and medical institutions should establish drug management system with a clear division of power and responsibility to relieve the drug shortages.

Objective To investigate the gene defects of a pedigree with inherited coagulation factor Ⅺ (FⅪ) deficiency by analyzing its phenotype and molecular genetic characteristics. Methods A pedigree with inherited FⅪ deficiency was enrolled in this study. The activated partial thromboplastin time (APTF), prothrombin time (PT), FⅪ activity (FⅪ: C) and FⅪ antigen (FⅪ: Ag) were determined for phenotype diagnosis. Fifteen exons and their flanks of F11 gene from the proband's genomic DNA were amplified by polymerase chain reaction (PCR), and the PCR products were directly sequenced to analyze the F11 gene mutation. The PCR products amplified from genomic DNA from the proband, her parents and 100 healthy donors were digested with restriction enzyme BssSI to exclude gene polymorphism and confirm the mutation site. The cleavage site in the signal peptide was predicted by the SignalP software. Results The values of APTT, PT, FⅪ: C and FⅪ: Ag of the proband were 69.5 s, 12.3 s, 2.6% and 2.5%, respectively, indicating that this case was cross-reacting material (CRM) negative. The same values of healthy controls were 35 s, 13 s, 100% and 100%, respectively. As compared with Genbank AY191837 sequence, four variants in F11 exons were found. G3733C heterozygous mutation in exon 2 causod Gly to Arg substitution at-1 amino acid position in signal peptide (G-1R). The G3733C mutation in exon 2 introduced a new BssSI enzyme digestion site. Further analysis of the 100 randomly collected DNA samples from the normal population excluded the possibility of G3733C as a polymorphism. CI6642T heterozygous mutation in exon 8 introduced a premature stop codon at 263 amino acid position (Q263Term). Conclusions G-1R mutation and Q263Term compound heterozygous mutation in F11 gene are the mechanism of FⅪ deficiency for the proband. G-1R mutation is a novel F11 gene mutation causing inherited FⅪ deficiency.