高通量筛选JAK-STAT6信号传导通路抑制剂方法的初步建立

目的构建可用于高通量筛选JAK/STAT6信号传导通路抑制剂的工程细胞株,建立稳定可靠的筛选方法。方法利用基因重组和转染技术,将STAT6特异性识别启动子IgE基因序列和虫荧光素酶报告基因联合插入pCMV质粒,脂质体法转染至HeLa细胞,经潮红霉素B抗性筛选及报告基因检测,得到稳定表达虫荧光素酶的工程细胞株。通过优化溶剂DMSO浓度,IL4作用浓度及孵育时间等筛选条件,建立了可靠的筛选方法,并在此基础上对1600种化合物进行了筛选。结果建立的筛选方法稳定可靠,系统Z′因子达到0.64。通过对1600种化合物的筛选,得到3个抑制效果较理想的化合物并测得其IC50值。结论所建立的高通量筛选方法可用于JAK/STAT6信号传导通路抑制剂的筛选。     

脾脏肿瘤的诊断及治疗

 目的　探讨脾脏肿瘤的诊断及治疗方法。方法　回顾分析我院 1978年 7月至 1999年 6月手术切除的 2 5例脾脏肿瘤的临床资料。结果　脾脏肿瘤男女发病率相似 ,临床表现以左上腹不适、疼痛及左季肋部肿物多见 ,辅助检查B超诊断率为 6 8% ,CT诊断率为 10 0 % ,良性肿瘤以脾囊肿最常见 ,恶性肿瘤以脾淋巴瘤最常见 ,良性肿瘤的治疗采用脾切除术或脾部分切除术 ,恶性肿瘤行脾切除或联合脏器切除辅以化疗。结论　脾脏肿瘤术前诊断依据CT、B超联合检查 ,最后确诊需手术切除及病理诊断 ,脾脏肿瘤无论良恶性均需手术治疗     

Continuous Spinal Anesthesia/Analgesia for Perioperative Management of Morbidly Obese Patients Undergoing Laparotomy for Gastroplastic Surgery

Background: The authors determined prospectively the safety of continuous spinal anesthesia combined with general anesthesia and the efficacy of postoperative pain relief with continuous spinal analgesia for morbidly obese patients undergoing vertical banded gastroplasty. Methods: 27 patients (13 men, 14 women) with a mean body mass index (BMI) of 50.4 ± 7.8 and several co-morbidities were studied. All patients were anesthetized with the same anesthetic regimen, which included midazolam, fentanyl, propofol, muscle relaxants, N₂O, isoflurane and intrathecal bupivacaine. Postoperative pain relief was provided for 5 days and all patients received the same regimen, which included intrathecal bupivacaine, fentanyl and intravenous tenoxicam. The intrathecal analgesic regimen was administered continuously through a pump which had the facility of providing bolus doses when requested in predetermined lockout intervals. Intra-operative monitoring included hemodynamic and respiratory parameters. Additional postoperative monitoring included respiratory rate, degree of sedation, sensory level of anesthesia, motor response and intensity of pain. Results: Intraoperative anesthetic technique was safe and provided satisfactory results in the immediate postoperative period. Furthermore, the postoperative analgesia regimen provided effective analgesia in all patients.The mean doses of fentanyl and bupivacaine infused intrathecally for the first 24 postoperative hours were 14.1 ± 2.0 μg.h^-1 and 0.7 ± 0.1 mg.h^-1 respectively, while the requirements of anal gesia decreased progressively with time. The technique provided effective analgesia with low pain scores, which was reflected by ease in mobilizing and performing physical exercises with the physiotherapist. Only minor complications related to anesthesia and analgesia were encountered. Conclusion: To our knowledge, this technique of anesthesia and postoperative analgesia has not been described before in morbidly obese patients. This regimen merits further controlled trials to establish its place in the perioperative management of morbidly obese patients.     

体外培养小鼠耳蜗Corti器对庆大霉素的吸收

目的 观察庆大霉素-德州红耦联物在体外培养小鼠耳蜗Corti器的分布情况,比较不同时间庆大霉素吸收的差异.方法 分离小鼠耳蜗Corti器,体外培养,培养液中加有庆大霉素-德州红耦联物,分别于培养后3 h、6 h、12 h、24 h、48 h、3 d.4 d和7 d收集标本,荧光染色后行激光扫描共聚焦显微镜观察基底膜庆大霉素的分布情况.结果 小鼠耳蜗Corti器在培养3 h后即可见外毛细胞的纤毛和胞体两侧的胞膜有庆大霉素分布;随着培养时间的延长,庆大霉素在Corti器中的所有细胞均见分布,尤以外毛细胞明显,主要聚集在毛细胞顶端纤毛的下方;培养24 h后庆大霉素在外毛细胞的聚集达到最高峰,而在支持细胞未见明显的聚集;体外培养7 d后在毛细胞胞质中仍可见庆大霉素分布.结论 小鼠Corti器体外培养可用来动态检测庆大霉素在耳蜗细胞的吸收和聚集情况.     

Cytogenetic and molecular genetic analysis of tumorigenic human bronchial epithelial cells induced by radon alpha particles

To establish a cell culture model for lung carcinogenesis, independent populations of the human papillomavirus 18-immortalized human bronchial epithelial cell line BEP2D were treated with high linear energy transfer radon-simulated alpha-particles, expanded and xenotransplanted into Nu/Nu mice. Six independent cell lines were established from tumors that developed from three separate radiation treatments as follows: treatment (Tx) 1 (30 cGy--two doses), H2BT, Tx 2 (30 cGy-- single dose), R30T1L, R30T2 and R30T3L, Tx 3 (30 cGy--single dose), H1ATN and H1ATBA1. Cytogenetic analysis revealed common changes in all tumor lines: loss of the Y chromosome (ch), one of three copies of ch8, one of three copies of ch14, and one of two copies of ch4p16-pter and ch11p15-pter. Analysis of polymerase chain reaction-amplified short tandem repeats of informative loci confirmed the loss of chY in all lines and loss of heterozygosity (LOH) at eight loci spanning the length of ch8 in all lines from Tx's 1 and 2. Our data support previous studies indicating the presence of tumor suppressor genes on ch8. LOH also was confirmed on ch14 at locus D14S306 in all cell lines from Tx 2 and in one of two lines from Tx 3. This region, 14q12-q13, may contain changes in one of the five known somatostatin receptor genes (SSTR1). No LOH was detected at any of the informative loci tested for on ch4 or ch11.      

Model of normal prepubertal growth

Overheating may cause terminal apnoea and cot death. Rectal temperature and breathing patterns were examined in normal infants at home during the first 6 months of life. Twenty one infants had continuous overnight rectal temperature and breathing recordings for 429 nights (mean 20.4 nights, range 7-30) spaced over the first six months of life. Periods when breathing was 'regular' were directly marked on single night records. Sleep state was determined from respiratory variables. 'Regular' breathing was a reliable marker of 'quiet' sleep (specificity 93%). The duration of 'quiet' sleep increased from 6 to 22 minutes from two weeks to three months of age and then remained static, as did the proportion of sleep spent in the quiet phase (9% to 34%). Rectal temperature fell during 66% of quiet sleep and usually rose during rapid eye movement (REM) sleep. The drop in rectal temperature was maximal at the start of quiet sleep, whereas the maximum rise during REM sleep was reached after 10 to 15 minutes. Oscillations in rectal temperature are associated with changes in sleep and breathing state. The maturation of rectal temperature patterns during the first six months of life are closely related to a maturation of sleep state and breathing patterns.     

Feeding problems in young PKU children

Behavioural feeding problems were found to be more prevalent in a group of 15 PKU children aged 1-5 years when compared to non-PKU controls. The parents of PKU children identified poorer appetites ( p < 0.01), a more limited range of foods consumed ( p < 0.03) and more gastrointestinal symptoms such as vomiting and constipation ( p < 0.03) than control children. The children were slower to feed ( p < 0.03), were more likely to dislike sweet foods and some ate separately from the rest of the family at mealtime ( p < 0.03). The effects on normal feeding behaviour should be considered when advocating strict diet therapy for young PKU children.