2004年 · 第117卷第12期

Background Atrial fibrillation (AF) is a common arrhythmia associated with increased cardiovascular morbidity and mortality. This study was undertaken to analyze the epidemiological factors and evaluate the current status of treatment in patients with AF in Mainland China. Methods Retrospective analysis of hospital records were taken from patients with primary diagnosis of AF, discharged from January 1999 to December 2001. A total of 9297 cases (mean age 65. 5 years) with AF were enrolled from 40 hospitals in major parts of China. Results The percentage of hospital admissions with AF was gradually increased comparing to those of total cardiovascular admission during three years, with the average of 7.9 %. The cases distribution progressively rose with age. The causes and associated conditions of AF: advanced age 58. 1 %, hypertension 40. 3 %, coronary heart disease 34. 8%, heart failure 33.1 %, rheumatic valvular disease 23.9%, idiopathic AF 7.4%, cardiomyopathy 5.4%, diabetes 4. 1 %. The most common coexistence among these variables was advanced age with hypertension. Permanent AF almost accounted for half of these cases (49. 5 %), paroxysmal and persistent AF were 33. 7 %and 16. 7 %, respectively. Paroxysmal AF was mainly treated with rhythm control (56. 4 %) . However, 82. 8 %of patients with chronic AF had therapeutic strategy of rate control. Inpatients with persistent AF, the cardioversion had been attempted in cases more than 50 %, with only 31. 1 % of these patients who could maintain stabilized sinus rhythm. The prevalence of stroke in this group was 17. 5 %. In nonvalvular AF patients the risk factors that significantly associated with stroke included advanced age, history of hypertension, coronary heart disease and type of AF. Sixty-four point five percent of these patients received antithrombotic therapy with dominated use of antiplatelet agents. The long-term prevention with anticoagulants only accounted for 6.6%. In this investigation patients with antiplatelets as well as patients with anticoagulants showed significant lower stroke rate in comparison with those managed neither. However, the difference between antiplatelets and anticoagulants in terms of stroke rate was not significant. Conclusions Most epidemiological factors of AF from this group showed highly in accordance with those from the reports from other countries, such as age distribution, causes and associated conditions, type of AF, dominantly with approach of rate control. Both antiplatelet and anticoagulant treatments significantly reduced stroke rate. But there was no significant difference between these two kinds of treatments in reducing stroke rate. Chin Med J 2004; 117 (12): 1763-1767

Background Typical accessory pathways (APs) of Wolf-Parkinson-White syndrome have been widely discussed in recent decades. However, the characteristics of the special AP, Mahaim fibre, are not so clear. It is known that these fibres have antegrade conduction only, long conduction time, decremental node-like conduction and automaticity properties. This study was to elucidate the automaticity of Mahaim fibre and its response to effective ablation. Methods Thirteen patients with Mahaim fibre (ten atrioventricular and three atriofascicular accessory pathways) were subjected to electrophysiological study and radiofrequency ablation via catheter. The incidence and characteristics of anautomatic rtythm o riginating from Mahaim fibre were observed during the whole procedure, especially during radiofrequency current delivery. Results Repetitive and short-run automatic rhythm (rate: 65 - 72 beats per minute), with a QRS morphology similar to that of clinical pre-excited atrioventricular re-entrant tachycardia (AVRT), occurred in two patients during sinus rhythm. Conduction via Mahaim fibre was successfully eliminated by radiofrequency current. Fourteen applications of RF were associated with irregularly accelerated automatic tachycardia of Mahaim fibre (with a sensitivity of 78%), lasting for 1.2- 14 seconds. However, such automatic tachycardia of Mahaim fibre did not occur during 54 failed applications of radiofrequency current. Conclusions Mahaim fibre has the function of automaticity. The accelerated automatic tachycardia of Mahaim fibre occur red during radiofrequency catheter ablation can be used as a predictor for successful procedure. Chin Med J 2004; 117 (12): 1768-1771

Background Prehospital delay remains one of the main causes of reduced benefit of reperfusion therapy for patients with acute myocardial infarction (AMI). The largest proportion of prehospital delay involve s the interval between the onset of synptoms and the decision to seek medical treatment. The purpose of this study was to examine the factors associated with the extent of care-seeking delay in Beijing for patients with AMI. Methods A structured interview was conducted in 102 patients with AMI in eight hospitals in Beijing. Results The mean decision time inpatients with AMI was (204±43) minutes, and prehospital delay time was (311 ±54) minutes. Only 34 % of patients sought medical care within one hour and a further 36 %of patients presented to one of the eight hospitals within two hours after onset. Educational level, atypical presentation of AMI, and family members at the site where AMI occurred were associated with longer delay time in seeking medical assistance (P <0.05, respectively), whereas the intensity of chest pain was inversely related to patients' delay time (P <0. 01). Patients who perceived their family relationship as good, attributed their symptoms to AMI origin, knew the time- dependent nature of reperfusion therapy, or used emergency medical service tended to seek medical care in a more rapid manner (P <0. 05, respectively). Conclusions Patients with AMI in Beijing delay seeking medical care to a great extent. Health education to increase the level of awareness of the target population at increased risk of AMI, including patients and their family members, is probably beneficial to reduce patients' care-seeking delay. Chin Med J 2004; 117 (12): 1772-1777

Background Many patients requiring allogeneic hematopoietic stem cell transplantation (HSCT) do not have an human loukocyte antigen (HLA)-matched donor. Alternative donors, such as HLA mismatched family donors, are associated with higher rates of graft rejection and acute graft versus host disease(aGVHD) if T cells are not first depleted. We developed a new technique for HLA mismatched allogeneic HSCT using GCSF primed bone marrow plus GCSF-mobilized peripheral blood stem cells without ex vivo T cell depletion. Methods In this study, 58 patients, including 33 with high-risk or advanced leukemia, were transplanted with cells from an HLA-haploidentical family donor with 1-3 mismatched loci. After conditioning, patients received GCSF-primed bone marrow grafts that had not been depleted ex vivo of T cells, in combination with GCSF-mobilized peripheral blood stem cells, as well as GVHD prophylaxis. Result All patients achieved sustained, full donortype engraftment. The incidence of grade Ⅱ- ⅣaGVHD was 31.9%, including 3 patients with grade Ⅲ-Ⅳ aGVHD. The development of aGVHD was not associated with the extent of HLA disparity. Chronic GVHD was observed in 30 of 51 evaluable patients(65. 4 %) . Fourteen patients died among whom 7 died of recurrent disease and 7 of transplant-related complications. Forty-four of the 58 patients survived, and 42 remained disease free at the time of a median followup of 12 months (3. 5 to 39. 5 months) . The 2-year probabilities of disease-free survival were 74. 8 %and 69. 3 %for standard- and high-risk patients, respectively. Conclusion We developed a new method to use bone marrow from haploidentical family donors without ex vivo T cell depletion, in combination with GPBSCs, as a source of stem cells even in cases of HLA mismatched transplantation. Chin Med J 2004; 117 (12): 1778-1785

Background Urethral reconstruction for both congenital and acquired etiologies remains a challenge for most urologic surgeons. Tissue engineering has been proposed as a strategy for urethral reconstruction. The purpose of this study was to determine whether a naturally derived extracellular matrix substitute developed for urethral reconstruction would be suitable for urethral repair in an animal model. Methods A urethral segmental defect was created in 20 male rabbits. The urethral extracellular matrix, obtained and processed from rabbit urethral tissue, was trimmed and transplanted to repair the urethral defect. Then, the regenerated segment was studied histologically by haematoxylin-eosin staining and Van Gieson staining at 10 days, 3 weeks, 6 weeks, and 24 weeks postoperation. Retrograde urethrography was used to evaluate the function of the regenerated urethras of 4 rabbits 10 and 24 weeks after the operation. The uro dynamics of 4 rabbits from the experimental group and control group I were assessed and compared. In addition, 4 experimental group rabbits were examined by a urethroscope 24 weeks after the operation. Results At 10 days after operation, epithelial cells had migrated from each side, and small vessels were observed in the extracellular matrix. The matrix and adjacent areas of the host tissue were infiltrated with inflammatory cells. The epithelium covered the extracellular matrix fully at 3 weeks postoperation. Well-formed smooth-muscle cells were first confiirmed after 6 weeks, at which point the inflammatory cells had disappeared. At 24 weeks postoperation, the regenerated tissue was equivalent to the normal urethra. Urethrography and uro dynamic evaluations showed that there was no difference between normal tissue and regenerated tissue. Conclusions Urethral extracellular matrix appears to be a useful material for urethral repair in rabbits. The matrix can be processed easily and has good characteristics for tissue handling and urethral function. Chin Med J 2004; 117 (12): 1786-1790

Background Evidence for the importance of genetic factors in male infertility is accumulating. This study was designed to identify a novel testis-specific gene related to spermatogenesis by a new strategy of digital differential display (DDD) . Methods Based on the generation of expressed sequenced tags (ESTs), comparing the testis libraries with other tissue or cell line libraries by the DDD program, we identified a new contig of the ESTs which were derived from testis libraries and represented a novel gene. Multi-tissue RT-PCR was performed to analyse its tissue-specific expression. The full-length cDNA of the new gene was obtained using the BLAST program. Sequencing was performed and the result was analysed. Semi-quantitative RT-PCR and Northern blot analyseis of mRNA from differential normal tissues were performed to clarify the expression pattern of the new gene. The sequence of the opening reading frame was integrated into the pQE-30 vector expressed in Escherichia coil strain Ml 5 (pREP4). With IPTG induction, the target protein was detected. Results A full-length cDNA sequence of the new gene named SPATA12 (GeneBank accession number AY221117) in human testis was identified. SPATA12 was 2430 bp in length, located in chromosome 3p21. l-3p21.2. The sequence of the opening reading frame was 676 - 1248 bp, as was confirmed by RT-PCR and sequencing. The cDNA encodes a novel protein of 190 amino acids with a theoretical molecular weight of 20 417. 8 and isoelectric point of 5. 23. The sequence has no significant homology with any known protein in databases. Semi- quantitative RT-PCR and Northrn-blot analyses of multiple tissues showed that SPATA12 was expressed significantly in normal human testis. The expression recombinant of SPATA12 was constructed and a high level of the histidine-tagged fusion protein was obtained. Conclusions DDD can be confirmed by SPATA12 as a novel computational biology-based approach for identification of the testis-specific expression genes. SPATA12 may function as a testicular germ cell associated gene that plays some roles in spermatogenesis. Moreover, a great amount of SPATA12 protein could be obtained by the gene recombination technique, thus providing a reliable foundation for investigating the biological function of this new protein. Chin Med J 2004; 117 (12): 1791-1796

Background Mutations in GJB2 gene are a major cause of autosomal recessive congenital hearing loss and the cause in some rare cases of the autosomal dominant form. The purpose of this study was to investigate the frequency and the features of GB2 mutations in the Chine se patients with congenital sensorineural deafness. Methods Using PCR anplifying the entire coding region of GJB2 gene and direct DNA sequencing to analyze mutations in this gene among unrelated 69 cases with autosomal recessive congenital nonsyndromic deafness and 27 cases of dominant congenital deafness and 35 sporadic cases. We also detected mutations in GJB2 in 100 control subjects with normal hearing. Results 17. 4 %(12/69) of the probands in the autosomal recessive, 7. 4 %(2/ 27) of dominant families and 5.7%(2/ 35) of the sporadic congenital deafness patients had deafness-causing mutations in GB2, respectively. Nine types of the mutations in GB2 were detected in the recessive and sporadic group. They consisted of five types of polymorphism, and four types of deafness-causing mutation with homozygous 35del Gin 1 sporadic (1/35), and 235delC frame shift mutation in 1 sporadic (homozygotes) and 10 recessive patients (2 heterozygotes and 8 homozygotes), and homozygous 442G→missense mutation and homozygous 465T→A nonsense mutation in 1 different recessive proband, respectively. The 465T→A that related to recessive deafness was a novel mutation found by this study. The homozygous (10/69, 14. 5 %) and the heterozygous (2/69, 2. 9 %) GJB2 mutation in the recessive patients (12/69, 17. 4 %)and the homozygotes in the sporadic patient (2/35, 5.7 %) all had congenital severe to profound sensorineural hearing loss. 511G→A missense mutation and 299 - 300delATframeshift mutation were found intwo autosomal dominant congenital deafness families (2/27, 7.4 %) . The total mutation frequency of GJB2 was 12. 2 %(16/131) in the Chinese patients with congenital sensorineural deafness and 235delC was the most common deafness-causing mutation. Six types of mutation —5 types of polymorphism and 1 type of heterozygous deletion (235delC) mutation were found in the 100 control subjects. The carry rate of the most frequent type of mutation 235delC was 0. 5 %in the controls (1/200 alleles). 109G→A was the most frequent (15/ 100, 15 %) and 79G→A was the second common (8/ 100, 8 %) polymorphism in this population. Conclusions The general mutation rate of GJB2 is 12. 2 %(16/ 131) and the 235delC is the most common type of deafness-causing mutation in Chine se patients with congenital hearing loss. 465T →A nonsense mutation that is associated to autosomal recessive deafness is a novel mutation found by this screening. 511G→A and 299 - 300delAT mutations contribute to autosomal dominant hearing loss. The study further supports the view that the common types of mutation in GJB2 according to different ethnic background and that the mutation prevalence in the East Asian deafness population is lower than that in the white population. Chin Med J 2004; 117 (12): 1797-1801

Background Complement receptor type 2 (CR2) is the receptor for C3d and C3dg and for Epstein-Barr virus. The aim of our study was to explore whether CR2 can independently mediate the activation of mitogen-activated protein kinases (MAPKs, including ERK, JNK, and p38MAPK), and to highlight the molecular mechanism of CD4+ cell deletion in AIDS. Methods HOS cells (HOS-CR2) and HOS-CD4 cells (HOS-CD4CR2) stably expressing CR2 were established and then identified by FACS and We stern blotting. Activation and blocking tests of MAPKs were assessed by We stern blot. Cell proliferation was determined using Cell Titer Aqueous One Solution Reagent. Results FACS results showed that the positive rates of HOS-CR2 and HOS-CD4CR2 cells were greater than 96 %, and Western blot showed that the CR2 expression levels on HOS-CR2 and HOS-CD4CR2 cells were high. Activation and blocking tests of MAPKs (ERK, JNK, and p38MAPK) were carried out in HOS-CR2, HOS-CD4, and HOS-CD4CR2 cells. The activation of MAPKs in HOS-CR2 cells stimulated with PMA (100 ng/ml) and NHS (10%) was identical. The activation of MAPKs increased at 5 minutes, reached a peak at 10 minutes, and decreased to baseline within 30 minute s, all in a time-dependent manner; the activation of MAPKs was blocked by anti-CR2 McAb, PD98059 (inhibitor of ERK), and Wortmanin(inhibitor of PI-3K), respectively. In HOS-CD4 cells, MAPKs were activated by HIV-gp 160. In HOS-CD4CR2 cells, MAPK activation was induced by HlV-gpl60, 10%NHS, and HIV-gp160 + 10 %NHS;phosphorylation of p38MAPKwas dramatically induced by HIV-gp160 + NHS, and lasted for 1 hour. The cell proliferation results showed that HlV-gpl60 inhibited the proliferation of HOS-CD4 and HOS-CD4CR2 cells(P<0.01) and that NHS enhanced the effect of HIV-gp160 (P<0.01). Conclusions The activation of MAPKs is independently mediated by CR2 and that anti-CR2 McAb, PD98059, and Wortmanin block the activation of MAPKs, respectively. The results of the signal transduction and cell proliferation assays of HOS-CD4CR2 cells show that CR2 plays a role in the pathogenesis of HIV infection, especially in the inhibition of CD4+ cell proliferation. Chin Med J 2004; 117 (12): 1802- 1808

Background Inhibition of tumor growth by endostatin has been shown to be an effective strategy in cancer therapy in mice. However, its widespread application has been hampered by difficulties in a largescale production of the recombinant endostatin protein, rapid loss bio activity of the protein, and the cumbersome daily administration. These limitations could be resolved by in vivo delivery and expression of the endostatin gene. In this study, we observed the effect and advantage of endostatin gene therapy mediated by a recombinant adenoviral vector (Ad/hEndo) on the growth of hepatocellular carcinoma BEL-7402 xenografted tumors, comparison with recombinant endo statin protein. Methods Hepatocellular carcinoma BEL-7402 cells were inoculated subcutaneously in the flank of Balb/ c nude mice. Nine days after tumor cell inoculation, animals were given a cycle of four courses of intra-tumoral injections of Ad/hEndo of 5 X108 pfu (low-dose group) and 1 ×109 pfu (high-dose group) at intervals of six days, respectively. Recombinant human endostatin protein (rhEndo) was administrated daily subcutaneously at a dose of 10 mg ·kg-1 d-1 at a site nearby the tumor for ten days. The expression of endo statin mRNA in tumor tissue was analyzed by reverse transcription- polymerase chain reaction (RT-PCR) after Ad/hEndo injection. Dynamic changes of concentration of endostatin protein in tumor tissue were quantitated by enzyme-linked immunosorbent assay (ELISA). Results After 4 courses of treatment, the tumor growth rates of high-dose treated group with 1 X109 pfu of Ad/ hEndo were inhibited by 42. 26 % conpared with the Ad/ LacZ control group (P=0. 001) and by 46. 26 % compared with the NIH buffer control group (P=0. 003), respectively. However, in this study, Ad/ hEndo at low dose of 5 ×108 pfu failed to demonstrate significant inhibition of tumor growth, compared with control groups. After daily administration of recombinant human endostatin protein (rhEndo) for 9 days, the ratio of T/C (rhEndo group versus PBS group) was less than 47 %. However, two days after rhEndo treatment ceased, the ratio of T/C was more than 50 %. The peak of expression of endo statin mRNA in tumor tissue was at 2 or 3 days after administration intratumorally with Ad/ hEndo of 1 ×109 pfu and gradually dropped undetectable by day 7. Dynamic analysis of endo statin concentration in tumor tissue showed that the highest level of mRNA is up at the third day after injection, and dropped to basal level three weeks later. Conclusions Endo statin gene therapy mediated by a recombinant adenoviral vector had significantly inhibited the growth of hepatocellular carcinoma BEL-7402 xenografted tumors at a high dose of 1 ×109 pfu compared with other groups. The analysis of dynamic expression of endo statin in vivo indicated that Ad/ hEndo had acquired a high-level,relatively long-term expression in vivo and bio activity capability. Chin Med J 2004; 117 (12): 1809- 1814

Background Although there were several clinical and experimental studies discussing the pathogenesis of dural arteriovenous fistula (DAVF), the pathological process leading to intracranial DAVF so far remains unknown. In this study, we investigated the expression of vascular growth factors in order to elucidate the possible role of these factors for the development of DAVF and to study the biological activity of this unco mmonlesion. Methods We examined the histological features, proliferative and angiogenic capacities of the tissue specimens obtained from 6 patients who underwent surgery at our institution. Immunohistochemical staining for vascular endothelial growth factor (VEGF), its receptors Flk-1 and Flt-l, ephrin-B2, MIB-1 and proliferating cell nuclear antigen (PCNA) was performed using standard immuno histo chemical techniques. Results A positive immuno staining was found for all antibodies studied except MB-1, whereas nuclear endothelial expression of PCNA was observed in only 3/6 cases. VECF stained positive in all of the available specimens (6/6) . Flk-1 showed a positive immuno re action in only 2/6 cases and Flt-1 in 4/6 cases. Ephrin-B2 was expressed in the majority (5/6) of the cases. Conclusions These results support the hypothesis that DAVFs might be acquired dynamic vascular malformations with low biological activity. Vascular growth factors like VEGF and ephrin-B2 might play a pivotal role in the formation of DAVF. Chin Med J 2004; 117 (12): 1815-1820