Aim To study the inhibitory effects of F951,a novel bcl-2 antisense oligodeoxynucleotide,on expression of bcl-2,growth of tumor and survival time of nude mice transplanted subcutaneously with acute myeloid leukemia.Methods HL-60 cells with high expression of bcl-2 were proliferated in vitro.The models of the nude mice with HL-60 cells were established by subcutaneous transplantation with drugs directly injected.The effects of F951 and F951 with low dose Ara-c on growth of tumor and survival time of mice with tumor were observed.The expressions of bcl-2 mRNA in the tumors implanted were detected by fluorescent quantitation RT-PCR.The morphologic structure of tumor tissues was assayed by light microscope.Results After each group mice with tumors were treated for 14 days,the volume,the weight of tumor and the bcl-2 mRNA expression of tumor tissue were shown respectively as follows: NS control group(15.17?3.40)cm3、(12.69?0.92)g、9.79?104 Copies??g-1;FNS group(15.91?3.77)cm3,(12.38?1.21)g;8.31?104 Copies??g-1;Ara-C group(1.24?0.55)cm3,(2.32?0.49)g,2.59?104 Copies??g-1;F951 group(2.6?1.55)cm3,(3.53?0.67)g;1.01?103 Copies??g-1;F951+Ara-C group(0.62?0.48)cm3,(1.05?0.63)g,9.5?102 Copies??g-1.The data above showed that F951 could downregulate the expression of bcl-2 in nude mice with HL-60 cells xenograft and inhibit growth of tumor.The growth of tumor of F951 group was reduced,and the inhibitory rate was 72.18%,there was significant difference comparing control groups with NS and FNS(P

Aim To investigate whether F951,a novel Bcl-2 antisense oligodeoxynucleotide,down-regulates Bcl-2 expression in HL60 cells and inhibits HL-60 cells proliferation.Methods HL60 cells were cultured with F951 in variant doses.The proliferation of HL60 cells was assayed by MTT and Typan Blue exclusion test.Expression of Bcl-2 protein and its mRNA was measured by FACS and RT-PCR respectively.The apoptotic cells were detected by DNA ladder.Results After HL60 cells were treated with F951 in 5,10,20 ?mol?L~(-1) doses respectively for 1～5 days,they showed apparent inhibition of proliferation.With the improvement of the concentration of F951 and the prolongation of the time of treatment, F951 showed stronge effect in the aspect of inhibiting the HL60 cells proliferation.It was determined with MTT method that the inhibition rates of HL60 cells treated with 5,10,20 ?mol?L~(-1) F951 were 20.56%, 37.66%, 54.11% respectively. F951 significantly down-regulated the expression of Bcl-2 mRNA and protein in the HL60 cells.Typical DNA ladder was seen from gel electrophoresis. With the improvement of the concentration of F951,it showed more apparent effect in the aspect of inducing DNA ladder.Conclusion F951 can inhibit cells proliferation through down-regulating Bcl-2 gene expression and promoting cells apoptosis in HL60 cells.

Aim To observe the apoptosis of human leukemic cells induced by F951, a novel bcl-2 antisense oligodeoxynucleotide. Methods HL60 cells were cultured with F951 in variant doses. Apoptotic cells were detected by flow cytometry, DNA ladder, electron microscope observation and terminal-deoxynucleotidyl transferase mediated nick end labeling (TUNEL). Results After HL60 cells were treated for 48 hours, mitochondria apoptotic cells can be detected at the ratio of 3.00% in the untreated group, 13.57% in the FNS control group and 30.95%, 38.08%, 52.55% with 5, 10, 20 ?mol?L-1 F951 respectively; the ratio of cells with caspase activity was 0.08% in the untreated group, 0.14% in the FNS control group and 43.68%, 60.54%, 80.37% with 5, 10, 20 ?mol?L-1 F951 respectively. Typical DNA ladder was seen from gel electrophoresis in F951 treated groups, and more apparent effect in the aspect of inducing DNA ladder had been observed with the improvement of F951 concentration. Detected through TUNEL and electron microscope, apoptotic cells in untreated group and FNS control group can only be found by chance, but very commonly seen in F951 treated groups and more frequently with the improvement of F951 concentration. Conclusion F951 can induce cell apoptosis in HL60 cells. Such effect is achieved through the inhibition of bcl-2 gene expression by F951 which initialize apoptosis passageway in consequence.

Objective To investigate outcomes of transanal endoscopic microsurgery(TEM) for the treatment of rectal villous adenoma and early-stage rectal carcinoma.Methods A series of 31 patients with rectal tumors underwent TEM from November 1995 to December 2003.The operation was performed under general anesthesia.The patients were placed in a dependent position dictated by the location of the tumor.A special rectoscope was inserted into the anus with CO_2 insufflation to keep the rectum open.Under the stereoscope and laparoscopic-type instruments,the tumor was completely resected(submucosal or full-thickness excision) using a needle diathermy or a 5-mm ultrasonic dissector.The operative wound was closed with intra-lumen continuous sutures.Results The rectal tumor was completely removed with negative resection margins in all the 31 patients.The operating time was 45~220 min(mean,95 min) and the intraoperative blood loss was 0~180 ml(mean,40 ml).Complications included temporary flatus incontinence in 2 patients,acute retention of urine in 1 patient,exacerbation of chronic obstructive airway disease in 1,and secondary hemorrhage following Aspirin taking in 1.The postoperative pathological stages were pT_0 in 16 patients,pT_(is) in 2 patients,pT_1 in 7,pT_2 in 3,and pT_3 in 3.Follow-up checkups in the 31 patients for 2~92 months(mean,23 months) revealed no local recurrence.Conclusions TEM is a safe and effective minimally invasive surgical technique for the treatment of rectal villous adenoma and early rectal carcinoma.

Objective To detect the DMD gene mutation sites and the regions of breakpoints in Duchenne/Becker muscular dystrophy (DMD/BMD) patients in northern China. Methods Multiplex amplifiable probe hybridization (MLPA) was used to detect the mutation in 59 cases (51 cases with DMD and 8 with BMD) from northern China and dystrophin gene mutations in their parents. Results From northern China and dystrophin gene mutations 59 families found gene deletions in 33 cases of 59 DMD/BMD patients (55.9%), duplications in 6 cases (10. 2%) and point mutation in one case (1.7%). Intron 44 was most frequently affected (n = 13, 33.3%), followed by intron 50 (n = 11, 28.2%) and intron 45 (n=8, 20.5%). The novel mutations were identified, in two patients including two independent duplications carried by patient D1 149 and a point mutation [5208del(A)] carried by patient D1 65, which were not included in Leiden database. In addition, an exon 22 deletion was found in one patient, which was the first reported case in Chinese patients. Conclusions Deletions are mostly located in the hotspot between exon 45 and 50. Duplications mostly occurred in the 5' end of the gene. Intron 44 is the most frequently affected breakpoint in northern Chinese population.

The development of health-related quality of life (HRQL) instrument in traditional Chinese medicine (TCM) is increasing rapidly in China, but few studies focus on their necessity. This study explores the necessity of the development of TCM instruments from both theoretical and practice perspectives, and aims to explain whether the adoption of the cross-medical style is valid. Through theoretical analysis, both TCM and Western medicine instruments show the same objectives, whereas TCM instruments are more suitable for the Chinese social behavior, customs and expectations. In practical analysis, 47 TCM instruments were identified, among which 17 had 18 corresponding Western medical instruments. In the domains layer, except for physiological, psychological and social factors, TCM instruments focus more on the harmony between body and spirit, humanity and nature or human and society and the constitution, etc. In the facts layer, TCM instruments focus on the emotions, initiative social intercourse, TCM symptoms, diet, sleep, taste, feces and urine, etc. In addition, significant differences existed in the methods of information selection. There is no need to modify cross-medical style research except when TCM characteristic terms exist, but attention must be paid to the influence of culture in different areas. Therefore, the TCM instruments can resolve the limitations of the application of Western medical instruments to the Chinese setting, while also having remarkable abilities of information coverage and detection. Both forms of instruments have the capacity and requirement to inter-communicate with each other in order to serve the whole Chinese cultural system. Generally speaking, there is no need to modify the instruments in cross-medical style research. But this point requires further demonstration in the rigorous designed clinical trials.

Objective　 This paper was designed to investigate the feasibility of non-invasive prenetal diagnosis of Duchenne muscular dystrophy(DMD). Methods　 The nucleated red blood cells(NRBC) were separated with percoll using a discontinuous density gradient method. The cells were smeared on microscope slides using a cyto-centrifuge and then stained by Wright-Giemsa. NRBCs were detected and individually retrieved into glass capillary pipettes using a micromanipulator under microscopic observation. The whole genome of a single cell was amplified by improved primer extension preamplification(PEP). The procedures for making prenatal diagnosis of DMD and determining the origin of NRBCs proceeded at the same time using sex determination and linkage analysis of several STR loci of dystrophin. Genotypes were analyzed by amplifying the 9 STR fragments using fluorescence-PCR technique and NRBCs origin was further determined. Results　 A case of DMD in male fetus was diagnosed. Conclusion　 With the use of the method reported, the non-invasive prenatal diagnosis of DMD is possible.

Objective To construct the content of health education based on transcultural nursing in discharged patients who were acute myocardial infarction (AMI) and had different cultural backgrounds,and to instruct the nurses to choose and formulate the individual health education program and prescription according to patients' cultural background.Methods Twenty experts who worked in tertiary A hospitals for 15 years,had secondary senior positions,were bachelor degree or above in nursing,cardiovascular and social science were chosen and received the expert consultation of Delphi method through the self-designed framework consulting table with three-level core content of the health education.The items on the content of health education at every level were quantitatively analyzed.Results The Cr was 0.865,and the Cs was 0.840,and the Ca was 0.890.The kandall coefficients of concordace at every level were respectively 0.425,0.443,0.408,and chi-square test showed that the difference was statistically significant (P ＜ 0.01).The number of items on the core content of health education through two rounds of expert consultation was determined including 11 items Grade Ⅰ,42items Grade Ⅱ and 126 items Grade Ⅲ.Conclusions The expert opinion of items on the content of health education in discharged patients with AMI at every level is intensive,and the reliabihty of the result is high,and which provides the scientific basis for carrying out effective health education in discharged patients who are AMI and have different cultural backgrounds.

OBJECTIVETo investigate the therapeutic effect of nerve growth factor (NGF) on changes of myelin basic protein (MBP) and functional repair of sensory and motor nerve following sciatic nerve injury.

METHODSThe sciatic nerves of rats were injured by sectioning with shaver,and divided into 3 groups: NGF group (Group A), group of normal saline solution (Group B), untreated group (Group C). The time point of observation was at the 4th week after operation. Sensory evoked potential (SEP) and motor evoked potential (MEP) were detected by Model WD-4000 nerve potential working diagnosis system. Immunohistochemical analysis was used for identification of MBP.

RESULTSThe latency of SEP in the Group A at the 4th week after operation was shorter than that in the Group B (P<0.05). The MEP was elicited in 76% of the Group A and was higher than that in the Group B. Results of immunohistochemistry showed that there were less MBP-positive cells in the Group A than in the Group B in one and four weeks respectively.

CONCLUSIONSNGF can improve the conductive function of injured peripheral nerve and facilitate regeneration of nerve.

Animals ; Evoked Potentials ; Female ; Immunohistochemistry ; Myelin Basic Protein ; metabolism ; Nerve Growth Factor ; pharmacology ; Peripheral Nerve Injuries ; Peripheral Nerves ; metabolism ; Rats ; Rats, Wistar ; Sciatic Nerve ; injuries ; metabolism

OBJECTIVETo explore the molecular pathogenesis and clinical phenotypes in 10 probands with inherited fibrinogen (Fg) deficiency.

METHODSThe diagnosis of hereditary Fg deficiency was validated by prothrombin time (PT), thrombin time (TT), Fg activity (Fg:C) and Fg antigen (Fg:Ag) in plasma. All of the exons and their flanking sequences of the Fg gene were analyzed by direct sequencing. Detected mutations were confirmed by reverse sequencing.

RESULTSThe ranges of Fg:C and Fg:Ag in the 10 probands were 0.52-0.91 g/L and 0.62-2.98 g/L, respectively. Five of the probands had type I disorders, and 5 had type II disorders. Seven point mutations were identified, among which 6 have located in the D region. γThr277Arg, γAsp316His, γTrp208Leu and Lys232Thr were novel mutations, and αArg19Ser was first reported in Chinese. Four probands had the same mutation site (γArg275). As to the clinical manifestation, probands with type I disorders were asymptomatic or with mild or medium symptoms, while those belonged to type II disorders had moderate or serious symptoms. Two probands have carried an Arg275Cys mutation but had different clinical manifestations.

CONCLUSIONMutations of the Fg gene seem to aggregate to the D region of FGG in our region, and Arg275 is a common mutation. However, no correlation has been found between the mutation site and clinical manifestations.

Adolescent ; Adult ; Afibrinogenemia ; blood ; classification ; genetics ; Base Sequence ; Child ; DNA Mutational Analysis ; methods ; Exons ; genetics ; Family Health ; Female ; Fibrinogen ; genetics ; metabolism ; Genotype ; Humans ; Male ; Middle Aged ; Mutation, Missense ; Partial Thromboplastin Time ; Phenotype ; Point Mutation ; Polymerase Chain Reaction ; Prothrombin Time ; Thrombin Time ; Young Adult