Drug Resistance ; Hormones ; pharmacology ; Humans ; Membrane Proteins ; genetics ; Nephrotic Syndrome ; genetics

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.

Objective To explore the value of 64-slice spiral CT angiography in diagnosis of pulmonary arteriovenous fistulas(PAVF).Methods 12 patients with suspect PAVF were performed by enhanced scan with 64-SliceCT.Two-dimensional and three-dimensional reformation were performed in all cases including coronal multiplanar reconstruction(MPR),maximum intensity projection(MIP),volume rendering(VR).Resnlts 12 cases were diagnosed as PAVF by 64-SliceCT angiography.Single PAVF in 7 cases,Multi-PAVF in 5cases.We found 20 lesions in 12 patients,Except direct multi-pulmonary arteriovenous fistulas in the area of near the heart in one case.Conclusion 64-slice spiral CT angiography is a useful and noninvasive means to identify PAVF and is also useful for therapeutic planning and postoperative follow-up.

OBJECTIVETo observe the effect of Qiju Dihuang Pill (QDP) on changes of Chinese medical syndrome types in pregnant women of Gan-Shen yin deficiency syndrome (GSYDS), and to explore the correlation between imbalanced cytokine levels and GSYDS.

METHODSThis was a random controlled trail. A total of 163 pregnant women of GSYDS at 12 -16 gestational weeks were randomly allocated into the experimental group (86 cases) and the control group (77 cases). Patients in the experimental group took QDP for 2 -4 weeks. Changes of Chinese medical syndrome types and serum interferon-gamma (IFN-gamma) and interleukin-4 (IL-4) levels were observed and compared between the two groups before and after treatment.

RESULTS(1) Totally 41 patients (47.7%) in the experimental group were transformed to normal Chinese medical syndrome type. In the same period of the follow-ups, 9 patients (11.7%) in the control group were transformed to normal Chinese medical syndrome type, showing statistical difference (P < 0.05). (2) In the experimental group, the serum level of IFN-gamma and the ratio of IFN-gamma/IL-4 in the peripheral blood were obviously lower after treatment than before treatment (P < 0.01), and obviously lower than those in the control group (P < 0.01). The level of IL-4 after treatment in the experimental group was higher than that before treatment, and also higher than that in the control group, but with no statistical difference.

CONCLUSIONSThese results indicated that there was imbalanced IFN-gamma/IL-4 ratio in the peripheral blood of pregnant women of GSYDS. QDP might play a role in immunoregulation by affecting the IFN-gamma level.

Adult ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Humans ; Interferon-gamma ; blood ; Interleukin-4 ; blood ; Medicine, Chinese Traditional ; Phytotherapy ; Pregnancy ; Yin Deficiency

Adolescent ; Anti-Arrhythmia Agents ; adverse effects ; therapeutic use ; Arrhythmias, Cardiac ; drug therapy ; physiopathology ; Child ; Child, Preschool ; Female ; Follow-Up Studies ; Humans ; Infant ; Male ; Sotalol ; adverse effects ; therapeutic use ; Treatment Outcome

Objectives To investigate and evaluate the preventive and therapeutic effects of celecox ib,a selective cyclooxygenase-2(COX-2)inhibitor,on multiple colorectal adenorna and compare it with aspirin.Methods Ninty-six patients with colorectal multiple adenoma were randomly divided into A,B and C groups.Adenomas in all patients were removed with high-frequency eleetrocoagulation,electroexci- sion or argon plasma coagulation(APC)under colonoscopy.Then,group A were administered celecoxib 200 mg twice daily,group B aspirin 50 mg twice daily,group C served as control.Colonoscopy was per formed every 6 months in the first year,and every year in order to observe and evaluate the recurrence rate of adenoma and the side effects after the treatment.Results Twenty-seven patients in group A,26 pa- tients in group B and 27 patients in group C had completed the treatment.At the end of the treatment, on PP/ITT analysis,the cure rate of the eolorectal adenoma were 84.4%/100% ,78.1%/96.2% and 75.0%/88.9% in group A,B and C,respectively.During the first year of follow-up,there were 1 ,1 and 6 cases which were found recurrences of the adenomas in group A,B and C,respectively.The recurrence rates of coloreetal adenomas in group A(3.7%)and group B(4.0%)were significantly low er than that in group C(24.0%) (P＜0.05 and＜0.05,respectively).At the end of follow-up,the total recurrence rate of colorectal adenomas in group A(14.80%)and group B(19.2%)were significant- ly lower than that in group C(46.2%)(P＜0.05 and＜0.05).While the side-effective rate regroup A (3.3%)was significantly lower than that in group B(22.5%)(P＜0.05).Conclusions After re- section of the multiple colorectal adenomas,both the selective inhibitor of COX-2,celeeoxib and the non- selective inhibitor of COX-2,aspirin,may reduce its recurrence rate,but the former has a good tolerance and lower side-effects.

Objective To investigate the antitumor therapeutic effect of combined therapy of magnetic induction heating by nano-magnetic particles, herpes simplex virus thymidine kinase gene(HSV-tk suicide gene) and internal radiation in mice bearing MCF-7 breast carcinoma. Methods The transfection reagents, plasmids heat shock protein-HSV-tk (pHSP-HSV-tk), ferroso-ferric oxide nano-magnetic fluid flow and 188Re-ganciclovir-bovine serum albumin-nanopaticles (GCV-BSA-NP) were prepared. The heating experiments in vivo were carried out using ferroso-ferric oxide nano-magnetic fluid flow. Sixty mice tumor models bearing MCF-7 breast carcinoma were established and randomly divided into six groups. Group A was the control group, B was gene transfection therapy group, C was hyperthermia group, D was gene transfection therapy combined with radionuclide brachytherapy group, E was gene therapy combined with hyperthermia group, and F was gene therapy, hyperthermia combined with radionuclide brachytherapy group. The tumor growth, tumor mass and histopathological changes were evaluated. The expression of HSV-tk in the groups of B, D, E and F was detected by RT-PCR. Poisson distribution and one-way analysis of variance (ANOVA) were used for statistical analysis by SPSS 10.0 software. Results In the animal heating experiments, the temperature of tumor increased up to 39.6 ℃, 43.2 ℃, and 48.1 ℃ quickly with different injected doses (2, 4 and 6 mg respectively) of nano-magnetic particles and maintained for 40 min. The temperature of tumor tissue reduced to 36.8 ℃, 37.5 ℃ and 37.8 ℃ in 10 min when alternating magnetic field (AMF) stopped. The tumor mass in Groups C ((452.50 ±30.29) mg), D ((240.98 ±35.32)mg), E((231.87 ±27.41) mg) and F ((141.55 ±23.78) mg) were much lower than that in Group A ((719.12±22.65) mg) (F=800.07, P＜0. 01), with the most significant treatment effect in Group F.The tumor mass in Group B((684.05 ±24.02) mg) was higher than that in Group D (t =32. 805, P ＜0. 05). Semi-quantitative RT-PCR analysis showed that the expression of HSV-tk in Groups B and D (0.33 ±0. 13 and 0. 46 ±0.12) was significantly different from that in Groups E and F (0.66 ±0.13 and 0.74 ±0. 11)(F = 21. 573, P ＜ 0.05). Conclusion Combined use of hyperthermia, gene therapy and radionuclide brachytherapy could effectively depress the growth of MCF-7 breast carcinoma, thus possessing treatment potential for this tumor.

The aim of this study was to investigate the origin of bone marrow-derived mesenchymal cells in 34 patients who had received a sex-mismatched hematopoietic stem cells transplant (HSCT). The mesenchymal stem cells (MSC) from 34 patients were collected for test. The different passage MSC of patients were analyzed by flow cytometry (FCM) to reveal the cell surface antigen expression. The polymerase chain reaction (PCR) analysis of the amelogenin (AMEL) genes was used to detect donor cells from host cells. The cultured MSC were stained by fluorescence in situ hybridization (FISH) probes for chromosomes X (Xp11.1-Xq11.1) and Y (Yq12) to distinguish donor cells from host cells. The results indicated that 31 out of 34 cases showed the confluent stroma and 24 out of these 31 cases were successfully passaged for more than 5 generations which could be used for PCR and FISH analysis. According to FCM results the number of CD14+CD45+ cells, which was regarded as monocyte/macrophage from the cultured MSC (passage 5) was less than 0.04%. In PCR assay, the marrow-derived MSC from the passage 5 were found to be of host origin. FISH assay demonstrated that marrow-derived MSC from the passage 5 were found to be of host origin up to 100%. It is concluded that the origin of MSC in bone marrow of patients after allogeneic stem cell transplantation was confirmed to be derived still from the recipients their own.
Adolescent ; Adult ; Bone Marrow Cells ; pathology ; Cells, Cultured ; Chromosomes, Human, X ; genetics ; Chromosomes, Human, Y ; genetics ; Female ; Hematologic Neoplasms ; therapy ; Hematopoietic Stem Cell Transplantation ; Humans ; In Situ Hybridization, Fluorescence ; Leukocyte Common Antigens ; analysis ; Lipopolysaccharide Receptors ; analysis ; Male ; Mesenchymal Stromal Cells ; pathology ; Middle Aged ; Peripheral Blood Stem Cell Transplantation ; Transplantation Conditioning ; Transplantation, Homologous

The aim of this study was to investigate the expanding capacity of bone marrow-derived mesenchymal stem cells (MSCs) in 34 patients who received a marrow and/or peripheral blood stem cells transplant (SCT). Marrow samples were obtained from iliac crest aspirates of healthy individuals (normal controls) and patients for the isolation, purification, and expansion of MSCs. The different passage MSCs of patients were analyzed by flow cytometry (FCM) to reveal the cell surface antigen expression. The expanding function of MSCs from patients after SCT, which might be affected by cytotoxic therapy in the conditioning regimen, colony-forming unit-fibroblast (CFU-F), confluent time, and passage number of the culture were measured, and then compared with those in the normal controls. At the same time, the numbers of colony-forming unit of hematopoietic progenitor were detected and compared with normal controls. In addition, CFU-F, confluent time, and passage number of MSCs were compared between group of BMSCs plus PBSCs co-transplanted patients and group of BMSCs plus PBSCs with the second donor umbilical cord blood cells (UBCs) co-transplanted patients. The results indicated that a confluent monolayer of stroma cells was generated in 31 out of the 34 cases (91.1%), a subconfluent monolayer was generated in one case (2.9%), no adherent stromal layer was generated in 2 cases (5.9%). As compared with the normal controls, the time generating a confluent layer of stroma cells from primary MSCs of patients was longer significantly, and the passage number and CFU-F of MSCs of patients in vitro was less than that of normal controls significantly. Compared with the group of BMSCs plus PBSCs co-transplanted patients, the confluent time of MSCs in group of BMSCs plus PBMSCs with UBCs co-transplanted patients was shorter, the passage number and CFU-F count in this group were higher. It is concluded that the MSCs of patients after HSCT are damaged, and the co-transplant of BMSCs and PBSCs plus UBCs can partially improve in vitro expanding capacity of MSCs from patients.
Adolescent ; Adult ; Bone Marrow Cells ; pathology ; Cell Proliferation ; Cells, Cultured ; Female ; Fetal Blood ; cytology ; transplantation ; Hematologic Neoplasms ; pathology ; therapy ; Hematopoietic Stem Cell Transplantation ; Hematopoietic Stem Cells ; cytology ; Humans ; Male ; Mesenchymal Stromal Cells ; pathology ; Middle Aged ; Peripheral Blood Stem Cell Transplantation ; Young Adult

To study feasibility of HLA-DRB1 matching by using denatured high performance liquid chromatography (DHPLC), 20 pairs of DNA samples from donors and recipients of hematopoietic cell transplantation (HCT) for DRB1 matching and 2 pairs of samples from donors and recipient of HCT for DRB1 mismatching were studied by DHPLC and PCR-SSP. After being amplified and annealed slowly to produce heteroduplex, PCR products for exon 2 of DRB1 were detected by DHPLC to find matched or mismatched peaks in chromatogram. The results showed that DHPLC and PCR-SSP were consistant with matched or mismatched HLA-DRB1 typing. The results of DHPLC and PCR-SSP for matching were compared by using kappa test (kappa = 0.776, P = 0.00), which suggested DHPLC for HLA-DRB1 matching was in agreement with PCR-SSP. In conclusion, DHPLC for HLA-DRB1 matching is economic and convenient, moreover, will not be affected by unknown genes in HLA-DRB1 locus.
Base Sequence ; Blood Donors ; Chromatography, High Pressure Liquid ; methods ; Feasibility Studies ; HLA-DR Antigens ; genetics ; immunology ; HLA-DRB1 Chains ; Hematopoietic Stem Cell Transplantation ; Histocompatibility Testing ; methods ; Humans ; Molecular Sequence Data ; Polymorphism, Genetic ; genetics