L-[S-11C-甲基]-蛋氨酸的快速制备和质量控制

利用回旋加速器轰击产生的11C-CO2为合成原料,甲醇化后获得反应活性很强的甲基化前体11C-CH3I,该前体再与L-高胱氨酸硫内酯在常温下反应,快速制备后获得11C标记的蛋氨酸(11C-MET).实验同时对30批次产品进行质量控制指标检测.结果表明从11C-CH3I到11C-MET合成时间为2min,合成效率为85%,产品放射化学纯度大于99%;质量控制指标合格.     

正电子发射体层摄影示踪剂^11C-乙酸的合成改进及其生物分布

目的建立快速、有效合成正电子发射体层摄影（PET）示踪剂11C-乙酸的方法并研究其生物分布,满足临床PET的用药需要。方法以^11C-CO2为原料,与CH3MgBr反应后再用17%的H3PO4酸解,产物通过加热的方法蒸出,同时通过控制^11C-CO2的释放速度和适当延长^11C-CO2与CH3MgBr的反应时间,提高反应产率。小鼠给药后不同时间处死,分别取不同器官称重并测放射线计数。结果从加速器生产出^11C-CO2到产物^11C-乙酸的蒸出完毕,需15min,放化纯度大于98%,化学纯度大于99%,平均产率为34.6%（未经衰变校正）。注射^11C-乙酸后,放射线主要集中在肝脏及肾脏。结论改进反应条件后的合成方法具有快速、高效的特点,为PET日常耗材节省了成本,提高了用药的质量。     

医专学生考试焦虑状况调查

目的了解医学生考试焦虑状况，为降低学生焦虑水平提供理论依据。方法采用Zung氏焦虑自评量表（SAS）分别在考试前后对医专学生进行无记名问卷调查。结果①考试前学生焦虑状态的发生率为21．92％，而考试后为11．98％，两者之间差异显著（χ^2=13．414，P〈0．001）；②考前男生焦虑状态的发生率占其人数的29．29％，女生占14．69％。两者之间差异显著（χ^2=12．791，P〈0．001）；考后男女生焦虑状态的发生率无显著差异（χ^2=1．172，P=0．279）。③不同年级焦虑状况比较，考试前一年级焦虑状态的发生率占其人数的11．76％，二年级学生占29．87％，三年级学生占22．69％．3个年级之间差异显著（χ^2=13．99，P〈0．001）；而考试后3个年级焦虑状态的发生率之间无差异显著（χ^2=1．413，P=0．493）。结论医学生在考试前有强烈的焦虑反应。提示学校要采取综合措施以降低学生考试焦虑水平。     

^11C-胆碱在线自动合成和生物体内分布

目的 研究[^11C-甲基]胆碱在线自动化合成的方法，并初步应用于临床试验．方法 ^11C-胆碱柱色层法在线制备，以Wistar大鼠研究动物体内生物学分布．结果 柱色层制备的^11C-胆碱放化纯度大于99％，合成效率为85％，合成时间11 min．动物分布表明：放射性主要分布于肾、肝和肠道，血液清除较快。结论 柱色层法制备^11C-胆碱时间短，效率高，操作简单。     

T淋巴细胞亚群检测在肾移植排斥反应与环孢霉素A中毒鉴别诊断中的应用

目的探讨外周血淋巴细胞亚群检测在人类同种异体肾移植术后急性排斥反应与免疫抑制剂环孢霉素A（cyclosporine A，CSA）中毒的诊断与鉴别诊断中的价值。方法采用四色流式细胞技术对26例肾移植术后肾功能正常、11例急性排斥反应、10例环孢霉素A中毒患者外周血淋巴细胞亚群中CD3^＋、CD3^＋CD4^＋、CD3^＋CD8^＋细胞的百分比进行检测。结果肾功能正常组、急性排斥反应组与环孢霉素A中毒组外周血淋巴细胞中CD3^＋细胞的百分比分别为71．83％±9．65％、73．29％±8．85％、72．06％±12．04％，3组比较差异无统计学意义；CD3^＋CD4^＋细胞的百分比分别为38．69％±9．21％、49．58％±8．41％、40．15％±9．98％，急性排斥反应组与正常组和CSA中毒组比较差异有显著统计学意义（P〈0．01）；CD3^＋CD8^＋细胞百分比分别为29.28％±9．02％、19．18％±5.35％、30．86％±9．19％，急性排斥反应组与正常组和CSA中毒组比较差异有显著统计学意义（P〈0．01）；CD3^＋CD4^＋／CD3^＋CD8^＋分别为1．76±0．97、2．92±0．71、1．81±0．92，急性排斥反应组与正常组和CSA中毒组比较差异有显著统计学意义（P〈0．01）。结论检测外周血淋巴细胞亚群的变化，对肾移植术后急性排斥反应和环孢霉素A中毒有鉴别诊断的价值。     

小剂量131I治疗甲亢的临床评价

为临床研究降低^131I治疗甲亢患者药物性甲减的发生，于1993—1995年采用小剂量^131I多次给药的方法．每次剂量不大于3mCi(111MBq)^131I，治疗甲亢300例。经7—9年随访，发现药物性甲减5例，发生率为1．66％。因此采用小剂量^131I给药法，可明显降低药物性“甲减”的发生，对于提高患者的劳作能力和生活质量有其显著效果。     

儿童重型β-地中海贫血同胞脐血移植后的免疫重建

目的分析儿童重型β-地中海贫血同胞脐血移植后的免疫重建情况,评价同胞脐血移植治疗重型β-地中海贫血的效果。方法11例患者移植后均为稳定骨髓植入,分别于移植后1、3、6、12、18～24、36～48个月检测患者外周血T淋巴细胞亚群（CD3^＋细胞、CD3^＋CD4^＋细胞、CD3^＋CD8^＋细胞）、B淋巴细胞（CD19^＋细胞）和自然杀伤细胞（CD3-CD56^＋NK细胞）数量,动态分析脐血移植后患者的免疫重建。结果14例患者脐血移植后11例获得植入,植入率为78．6％。11例患者移植后中性粒细胞绝对计数≥0．5×10^9／L、血小板计数≥20．0×10^9／L和≥50．0×10^9／L的中位时间分别为17、48、53d。总淋巴细胞和T淋巴细胞亚群在12个月内均恢复正常;B细胞和NK细胞分别在6个月和3个月内恢复正常。B细胞和NK细胞的恢复比T细胞快。11例患者发生急性移植物抗宿主病（GVHD）,其中I度急性GVHD9例（81．8％）,Ⅱ度急性GVHD2例（18．2％）。结论同胞脐血移植是根治重型β-地中海贫血的有效方法。     

9525例中孕期母血清学产前筛查效率评价分析

目的分析在严格质量控制的条件下,中孕期母血清学产前筛查效率及临床价值。方法通过规范的筛查前孕妇信息采集、实验中的质量控制以及筛查后高风险孕妇的召回与随访,回顾性分析2008年1月至2013年9月间进行中孕期血清二联筛查的9525例孕妇的筛查与诊断结果。结果9525例孕妇中筛查高风险589例。其中21-三体综合征高风险478例（确诊11例）,漏诊3例,检出率78．57％,假阴性率4．66％,阳性预测值2．42％。18-三体高风险24例（确诊2例,无漏诊）,检出率100％。ONTD高风险11l例（确诊3例）,检出率100％。筛查阳性孕妇检出胎儿为其他染色体异常者5例。结论本研究通过筛查实验前、中、后各个环节严格规范的全程质量控制,获得了较好的目标疾病的检出率,说明了筛查工作多环节的质量控制、后续产前诊断服务以及随访等工作,对于全面、准确地评价和分析总体筛查效率与效益的重要性。     

脐血CD34+细胞体外定向诱导分化为T淋巴细胞的实验研究

目的：建立利用人造血干／祖细胞体外定向诱导分化为T淋巴细胞的方法，为研究T细胞生物学特性及细胞免疫提供技术平台。方法：MACS方法分离人脐带CD34^ 细胞接种到人胎儿胸腺基质单层细胞上，IMDM液体培养基含20％人AB血清并加入FL、IL-12、IL-7和IL-2细胞因子组合，于培养7、14、21、28、35、42天取非贴壁细胞利用流式细胞仪对细胞表型进行检测，并进行细胞形态学分析。结果：2周后，CD4^ CD8^ 非成熟T淋巴细胞占细胞总数的0．3％-13．3％，4-5周CD4^ CD8^ T淋巴细胞达到高峰占16．6％-26．5％，且CD3^ CD4^ CD8^ 和CD3^ CD4^-CD8^ T淋巴细胞逐渐增多，6周后达26．5％～64．9％和11．6％-38．9％。培养成熟的T淋巴细胞经PHA IL-2刺激后瑞氏染色鉴定可见大原始淋巴细胞存在。结论：利用人脐血CD34^ 在体外人胎儿胸腺基质单层细胞上加FL、IL-12、IL-7和IL-2细胞因子组合条件下，可诱导分化出T淋巴细胞，并且培养的T细胞对有丝分裂素刺激有增殖反应。     

^131I治疗36例Graves病伴白细胞减少患者的临床疗效观察

为探讨^131I治疗对Graves病（GD）伴白细胞（WBC）减少患者或服用ATD后出现白细胞减少的GD患者的疗效,给予36例GD伴白细胞减少的患者^131I治疗,并分别测定患者治疗前和治疗后1个月、3个月、6个月的血常规、甲状腺激素水平等相关指标。结果显示^131I治疗3个月后,FT3、FT4水平明显降低,TSH升高,与治疗前比较有显著性差异（P〈0.05）;^131I治疗后3个月,外周血白细胞水平逐渐恢复,与治疗前比较有显著性差异（P〈0.05）。^131I治疗GD伴白细胞减少是一种安全可靠、疗效肯定的治疗方法,应尽早选用。     

11^C-蛋氨酸的体内生物学分布和初步临床应用

目的研究新型脑肿瘤显像剂：11^C-蛋氨酸（11^C-MET）的体内生物学分布，探讨11^C—MET的临床显像方法及其在脑肿瘤显像中的应用．方法测定11^C—MET在小鼠体内的分布，对健康志愿者和病理证实胶质瘤患者进行PET脑显像．结果小鼠体内分布实验和脑PET显像发现，11^C—MET在血液中清除较慢，在肿瘤内有较长的滞留时问，平台期位于30—45min之间，延迟相对有利于脑瘤的显像，其余放射性主要集中在消化系统．结论11^C—MDT有较高的肿瘤／脑比值，是理想的脑肿瘤显像剂．     

Genetic regulation of gametogensis: The receptor encoded by the human C-MET oncogene is expressed in testicular tissue and on human spermatozoa

Because of its distinctive ability to act as a mitogen, a mitogenand a morphogen, hepatocyte growth factor/ scatter factor (HGF/SF)has all the characteristics of a molecule able to function inregulatory networks of motility, such as the spermatogenic epithelium,and this through binding of its receptor p190^MET (C-MET). Inthis study we report the expression of C-MET in the human seminiferousepithelium and on spermatozoa from men being treated for infertilityand sperm donors. The presence of C-MET was demonstrated byimmunochemistry on the cell membrane of spermatogonia, spermatocytes,spermatids and on spermatozoa, whereas Sertoli cells and Leydigcells did not show expression. Comparison of C-MET expressionon spermatozoa of the 90% Percoll layer of subfertile patientsand donors revealed clearly two distinct groups (unpaired t-test,P < 0.001), whereas comparison of C-MET expression on spermatozoain the 47% Percoll layer was not significantly different betweenpatients and donors. In addition, there was a significant inversecorrelation between sperm concentration and the C-MET expressionof spermatozoa in the 90% Percoll layer (r = –0.80, 95%confidence interval, –0.92 to –0.55; P < 0.0001),but not with the C-MET expression of spermatozoa in the 47%Percoll layer. In conclusion, the presence of C-MET was demonstratedin the seminiferous epithelium and on mature and immature spermatozoa,indicating a role for this growth factor receptor in the differentiationand/or migration that occurs during human spermatogenesis. C-MET/male infertility/spermatozoa/testis     

Liquid chromatography-electrospray ionization mass spectrometric analysis of corticosterone in rat plasma using selected ion monitoring.

A simple and fast yet highly sensitive and specific method based on HPLC coupled to electrospray ionization mass spectrometry has been developed for the quantitation of corticosterone in rat plasma. After extraction of rat plasma (100 microl) with diethyl ether using 5-pregnen-3beta-ol-20-one-16alpha-carbonitrile (Sigma) as internal standard, HPLC was performed on a short C8 column (Zorbax-Eclipse, 50x4.6 mm I.D.) using a steep methanol-water gradient (methanol 54% to 90% in 6 min). Detection was performed on a single quadruple mass spectrometer in selected ion monitoring mode (m/z 369 for corticosterone and 364 for the internal standard). The detection limit of the assay was 9 fmol (3 pg) of corticosterone on column. In vitro data were subjected to curve fitting (cubic, r2=0.9999). Recovery of corticosterone after extraction ranged from 81 to 93%. The relative standard deviations for intra- and inter-assay precision ranged from 0.8 to 3.6% and 5.2 to 12.9%, respectively. Corticosterone did not undergo any appreciable degradation when stored in plasma at -20 degrees C for 2 months. The assay is routinely used in our laboratory to examine corticosterone levels as a marker of stress in rats and may also be used for the determination of 18-hydroxy-11-deoxycorticosterone.     

宫颈癌基因组HLA-Ⅰ类基因微卫星不稳定和杂合性缺失的研究

目的　探讨人白细胞抗原 Ⅰ类基因在宫颈癌中微卫星不稳定和杂合性缺失的情况 ,并构建宫颈癌基因组在该区域的精细缺失图谱。方法　应用聚合酶链反应 单链长度多态性 银染技术 ,对 3 0例宫颈癌活检标本进行杂合性缺失和微卫星不稳定的检测。结果　在 3 0例宫颈癌活检组织中 ,有 2 3例( 76.7% )存在有 1个或多个位点的杂合性缺失 ,微卫星位点C3 2 11的杂合性缺失频率最高 ,可达5 0 % ( 15 /3 0 )。微卫星不稳定的发生率为 66.7% ( 2 0 /3 0 ) ,其中位点D6S2 5 8发生微卫星不稳定频率最高 ,达 40 % ( 12 /3 0 )。结论　人白细胞抗原 Ⅰ类基因的微卫星不稳定和杂合性缺失在宫颈癌的发生和发展过程中起着重要作用 ,同时发现位点C12 5～C3 2 11之间是宫颈癌患者的一个最小的共同缺失区域 ,该区域可能存在有与宫颈癌相关的抑癌基因。     

Different fibre populations distinguished by their calcium transient characteristics in enzymatically dissociated murine flexor digitorum brevis and soleus muscles

Enzymatically dissociated flexor digitorum brevis (FDB) and soleus fibres from mouse were used to compare the kinetics of electrically elicited Ca²⁺ transients of slow and fast skeletal muscle fibres, using the fast Ca²⁺ dye MagFluo4-AM, at 20–22°C. For FDB two Ca²⁺ transient morphologies, types I (MT-I, 11 fibres, 19%) and II (MT-II, 47 fibres, 81%), were found, the kinetic parameters (amplitude, rise time, half width, decay time, and time constants of decay) being statistically different. For soleus (n = 20) only MT-I was found, with characteristics similar to MT-I from FDB. Correlations with histochemically determined mATPase, reduced nicotinamide adenine dinucleotide diaphorase and α-glycerophosphate dehydrogenase activities, as well as immunostaining and myosin heavy chain electrophoretic analysis of both muscles suggest that signals classified as MT-I may correspond to slow type I and fast IIA fibres while those classified as MT-II may correspond to fast IIX/D fibres. The results point to the importance of Ca²⁺ signaling for characterization of muscle fibres, but also to its possible role in determining fibre function.     

Rapid determination of gemifloxacin in human plasma by high-performance liquid chromatography-tandem mass spectrometry

A method was developed for the determination of gemifloxacin (I) in human plasma using high-performance liquid chromatography-tandem mass spectrometry. Prior to analysis, the protein in plasma samples was precipitated with acetonitrile containing [13C2H3] gemifloxacin (II) to act as an internal standard. The supernatant was injected onto a PLRP-S column without any further clean-up. The mass spectrometer was operated in positive ion mode, employing a heat assisted nebulisation. electrospray interface. Ions were detected in multiple reaction monitoring (MRM) mode. The assay requires 50 microl of plasma and is precise and accurate within the range 10-5,000 ng/ml. The average within-run and between-run coefficients of variation were <11% at 10 ng/ml and greater concentrations. The average accuracy of validation standards was generally within +/-7% of the nominal concentration. There was no evidence of instability of I in human plasma following three complete freeze-thaw cycles and samples can safely be stored for at least 6 months at -20 degrees C. The method proved very robust and was successfully applied to the analysis of clinical samples from patients dosed with gemifloxacin.     

Localization of the fast skeletal muscle troponin I gene (TNNI2) to 11p15.5: genes for troponin I and T are organized in pairs

We have localized the gene encoding the fast skeletal muscle isoform of troponin I (TNNI2) to 11p15.5 by PCR-based analysis of somatic cell hybrid panels: based on the Genebridge4 radiation hybrid panel, TNNI2 is coincident with the marker D11S922. The gene encoding the fast skeletal muscle troponin T gene (TNNT3) has been previously assigned to 11p15.5 suggesting that TNNI2 and TNNT3 may be closely linked. The overall location of genes encoding troponin I and T isoforms now reveals that they are organized at three loci each containing a troponin I/troponin T gene pair. This organization contrasts with all other sarcomeric protein genes and has implications for the evolution of these two gene families, for their regulation and for the analysis of mutations suspected to result in cardiomyopathy.     

Utilization of a sol–gel method for encapsulation of doxorubicin

The sol-gel pre-doping method was used to encapsulate doxorubicin in silica gels and optimum conditions of preparation of drug-loaded gel were established, ensuring both reproducible and effective results of quantitative encapsulation of doxorubicin and its gradual but complete release. Doxorubicin was encapsulated in polysiloxane polymers using the method based on sol-gel encapsulation without a catalyst, with an acid catalyst (HCl) and a base catalyst (NH₃). The time of gelation of the gel loaded with doxorubicin, encapsulation efficiency of the drug and the degree of release of the drug from the gel are all affected by the kind of catalyst (acidic or basic) or its absence at the gel preparation stage, and the temperature of the gelation process. The time of sol gelation when using the NH₃ or HCl catalyst was 9 days at 21°C, 2 days at 30°C and 1.5 days at 37°C, while for the gel prepared without a catalyst it was 90 days at 21°C, 75 days at 30°C and 70 days at 37°C. The efficiency of doxorubicin encapsulation was 99.5 ± 0.5% (w/w) for acid-catalyzed gel, 98.9 ± 1.01% (w/w) for base-catalyzed gel and 86.4 ± 11.6% (w/w) for non-catalyzed gel. A 100% (w/w) release of doxorubicin by diffusion through pores was found only in the case of base-catalyzed gel after a 140-h incubation time. For acid-catalyzed gel and non-catalyzed gel, the total amounts of released doxorubicin after 140 h of incubation were 3-5% (w/w) and 9-11% (w/w), respectively. The stability of doxorubicin encapsulated in the three kinds of gel matrices was found to be improved compared to the stability of a free form of the drug in solution.     

Utilization of a sol-gel method for encapsulation of doxorubicin

The sol-gel pre-doping method was used to encapsulate doxorubicin in silica gels and optimum conditions of preparation of drug-loaded gel were established, ensuring both reproducible and effective results of quantitative encapsulation of doxorubicin and its gradual but complete release. Doxorubicin was encapsulated in polysiloxane polymers using the method based on sol-gel encapsulation without a catalyst, with an acid catalyst (HCl) and a base catalyst (NH3). The time of gelation of the gel loaded with doxorubicin, encapsulation efficiency of the drug and the degree of release of the drug from the gel are all affected by the kind of catalyst (acidic or basic) or its absence at the gel preparation stage, and the temperature of the gelation process. The time of sol gelation when using the NH3 or HCl catalyst was 9 days at 21 degrees C, 2 days at 30 degrees C and 1.5 days at 37 degrees C, while for the gel prepared without a catalyst it was 90 days at 21 degrees C, 75 days at 30 degrees C and 70 days at 37 degrees C. The efficiency of doxorubicin encapsulation was 99.5 +/- 0.5% (w/w) for acid-catalyzed gel, 98.9 +/- 1.01% (w/w) for base-catalyzed gel and 86.4 +/- 11.6% (w/w) for non-catalyzed gel. A 100% (w/w) release of doxorubicin by diffusion through pores was found only in the case of base-catalyzed gel after a 140-h incubation time. For acid-catalyzed gel and non-catalyzed gel, the total amounts of released doxorubicin after 140 h of incubation were 3-5% (w/w) and 9-11% (w/w), respectively. The stability of doxorubicin encapsulated in the three kinds of gel matrices was found to be improved compared to the stability of a free form of the drug in solution.