流式细胞术动态检测肝大部切除后再生相关因子的实验研究

为探讨肝细胞生长因子(HGF)、转化生长因子-α(TGF-α)、转化生长因子-β(TGF-β)在肝再生中的作用及机制.应用流式细胞术(FCM)动态检测再生肝组织S期肝细胞百分含量及HGF、TGF-α、TGF-β的变化.结果表明HGF、TGF-α浓度在肝大部分切除6 h后达高峰,而TGF-β 24 h后缓慢上升.提示HGF、TGF-α、TGF-β分别通过不同的机制影响肝再生,HGF、TGF-α为正性调节因子,TGF-β为负性调节因子.     

肝胰十二指肠联合切除对残肝再生和肝细胞生长因子,转化生长因子α、β表达的影响

目的：探讨残肝再生和肝细胞生长因子(HGF)、转化生长因子α(TGF—α)和β(TGF—β)的表达。方法：30只SD大鼠随机分为2组：①组1为大部(68％)肝切除组，15只；②组2为大部(68％)肝叶切除加部分(50％)胰腺切除加近端十二指肠切除组，15只。每组3只大鼠，于术后12、24、48、72和168h分别处死并采集肝脏样本，计算肝再生率。切除的肝脏组织用RT—PCR技术检测HGF、TGF—α及TGF—β在不同时间点的表达，并与免疫组化法检测的肝细胞增殖细胞核抗原(PCNA)进行对比研究。结果：组1残肝再生率逐步增加，至术后第7天达最高水平；PCNA峰值在术后24h，随后逐步下降。HGF和TGF—α的表达在术后12h逐渐增强，24h达到高峰，随后逐渐下降，168h时达最低水平，而TGF—β的表达在48h时达高峰，然后下降。与组1相比，组2可显著减低残肝再生及PCNA峰值，并可显著减少肝细胞HGF、TGF—α及TGF—β的表达。结论：肝胰十二指肠联合切除可减低残肝再生反应，除了肝细胞自身分泌的HGF及冗F—Q等细胞因子外，起源于胰腺和十二指肠的其他细胞因子亦可对肝细胞的增殖起调节作用。     

肝切除术后肝再生调控机制的研究进展

肝脏是一种具有再生能力的特殊机体器官。部分切除肝、肝实质细胞的肝脏,剩余的非实质细胞及各种细胞因子、生长因子和各种类型的肝干细胞可迅速响应,促进肝脏恢复正常形态功能。本文首先介绍肝切除术后肝脏再生调控的效应细胞,主要包括肝细胞、肝窦内皮细胞、胆管上皮细胞和Kupffer细胞等。然后介绍肝脏再生大致分为三个阶段,其中参与这三个阶段调控的因子主要包括肿瘤坏死因子α、白细胞介素6、肝细胞生长因子与其受体CMet,表皮生长因子（EGF）与其受体EGFR和去甲肾上腺素,卵圆细胞,在肝切除后促进肝再生,促进肝脏疾病患者恢复身心健康。     

乙型肝炎肝乐宁治疗后血清甲胎蛋白的变化

动物胎肝中存在肝细胞生长刺激激素(HSS)及肝细胞生长因子(HGF),能刺激肝细胞DNA合成[1],促进肝细胞再生,降低转氨酶(ALT),增加枯否细胞的功能,促进病变细胞恢复.肝乐宁是从羊胎肝中提取的有效成分,临床应用中发现,它能使血中甲胎蛋白(AFP)升高.为此笔者对38例使用肝乐宁治疗的乙型肝炎患者,动态观察ALT、AFP与未用肝乐宁的28例病人(对照组)进行了对比,报告如下.     

阿德福韦对慢性乙型病毒性肝炎患者血浆细胞因子及肝纤维化指标的影响

目的 探讨阿德福韦对慢性乙型病毒性肝炎(乙肝)患者血浆细胞因子及肝纤维化指标的影响.方法 选取68例轻中度慢性乙肝患者,将其随机分为阿德福韦组和对照组,各34例.对照组给予常规保肝治疗,阿德福韦组在对照组常规保肝治疗的基础上加用阿德福韦酯(10 mg,1次/d)治疗6个月.分别在治疗前和治疗6个月后检测两组患者的肝功能[丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)]、血浆细胞因子[转化生长因子β1(TGF-β1)、肿瘤坏死因子α(TNF-α)、肝细胞生长因子(HGF)]和肝纤维化指标[透明质酸(HA)、Ⅲ型前胶原(PC Ⅲ)、Ⅳ型胶原(Ⅳ C)、层粘蛋白(LN)]的变化.结果 两组患者治疗前肝功能、血浆细胞因子水平及肝纤维化指标间差异无统计学意义(P＞0.05);而治疗后两组患者ALT、AST、TGF-β1、TNF-α、HA、PC Ⅲ、Ⅳ C、LN水平均较治疗前显著下降,HGF水平较治疗前显著升高,差异均有统计学意义(P＜0.05);且治疗后两组患者上述指标比较,差异亦有统计学意义(P＜0.05).结论 阿德福韦能促进HGF的表达,减少肝组织中TGF-β1、TNF-α分泌,进而抑制肝星状细胞活化和细胞外基质的合成,促进其分解,从而起到抗肝纤维化和减轻肝组织炎症的作用.     

半乳糖凝集素-3联合HGF诱导大鼠骨髓间充质干细胞分化为肝样细胞

目的 探索半乳糖凝集素-3(Gal-3)联合肝细胞生长因子(HGF)诱导大鼠骨髓间充质干细胞(BMSCs)向肝样细胞分化的可行性及最佳诱导条件,并对诱导后的细胞进行鉴定.方法 离心、贴壁培养分离大鼠BMSCs,流式细胞术鉴定表面标志.取第3代细胞进行分组诱导培养,A组:0 μg/mL Gal-3+ 20 ng/mL肝细胞生长因子(HGF);B组:0.1μg/mL Gal-3+ 20ng/mL HGF;C组:0.5 μg/mL Gal-3+20 ng/mL HGF;D组:1.0 μg/mL Gal-3+20 ng/mL HGF;E组:2.0 μg/mL Gal-3+ 20ng/mL HGF;阳性对照组:SD大鼠肝脏细胞IAR20.分别于诱导后7、14、21、28d时段进行细胞形态学观察、免疫荧光染色检测、实时荧光定量核酸扩增检测(Q-PCR)鉴定其诱导分化情况.结果 分组诱导培养后,各组BMSCs形态逐渐向肝样细胞转化,与大鼠原代肝脏细胞IAR20相似,其中28 d时间段,C组转化成IAR20相似细胞比例最高;各组甲胎蛋白(AFP)、清蛋白(ALB)荧光染色阳性率明显增高,并且在28 d时间点C组AFP、ALB的荧光染色阳性率最高;C组AFP、ALB mRNA表达明显增加,同时以28 d时间点表达量达到峰值.结论 Gal-3联合HGF能有效诱导大鼠BMSCs分化为肝样细胞,分化后的肝样细胞能分泌肝细胞特异性产物AFP、ALB,且其诱导效率与诱导培养时间和Gal-3浓度有关.     

转化生长因子-α及β与肝再生关系研究进展

肝脏具有强大的再生能力,其调控机制复杂.肝切除术对肝脏既有机械性损伤,又有缺血再灌注损伤,其再生过程是一个复杂的病理生理过程[1].非实质细胞在肝部分切除术后肝再生的应答过程中,起着重要的调节作用,调节着肝细胞的代谢和生长,而细胞与细胞之间相互作用的桥梁主要来自于细胞因子,细胞因子在肝再生过程中又起着非常重要的作用[2].其中转化生长因子α(TGF-α)和β(TGF-β)的调节作用越来越受到重视.     

门静脉栓塞及结扎联合肝细胞生长因子对肝纤维化大鼠肝再生的研究

目的 探讨门静脉栓塞及结扎联合肝细胞生长因子（hepatocyte growth factor,HGF）对肝纤维化大鼠肝再生的作用。方法 制备大鼠肝纤维化模型,将100只肝纤维化大鼠随机分为5组,术前对照组、门静脉栓塞组(A1组)、门静脉结扎组(A2组)、门静脉栓塞+肝细胞生长因子组(B1组)和门静脉结扎+肝细胞生长因子组(B2组）,每组20只。除术前对照组外以上各组栓塞或结扎大鼠门静脉右支。每组于术前及术后1、3、7及14 d分批处死大鼠,每次5只,检测各组时间点大鼠外周血清中丙氨酸氨基转移酶(alanine aminotransferase,ALT)、天冬氨酸氨基转移酶(aspartate aminotransferase,AST)的含量;称取各叶肝脏及全肝重量,计算术后不同时间点非栓塞及非结扎肝叶占整个肝脏的百分比;免疫组化检测增殖细胞核抗原（proliferating cell nuclear antigen,PCNA）与Ki- 67的表达。结果 术后各组ALT、AST呈一过性升高改变,表现为术后第1天达到高峰,B1与B2组术后第1、3、7天ALT及AST峰值分别较A1与A2组低(P <0.05)。各组大鼠非栓塞及非结扎侧肝重/全肝重百分比均随时间增加而增加,术后第7、14天B1组非栓塞叶肝重/全肝重较A1组高,B2组非结扎叶肝重/全肝重较A2组高(P<0.05)。免疫组织化学改变：A1与A2组、B1与B2组术后非栓塞及非结扎侧肝叶PCNA与Ki-67的表达较术前明显增加并于第3天达高峰(P<0.05),然后缓慢下降,第14天A1与A2组恢复至术前水平(P>0.05),而B1与B2组较术前仍明显升高(P<0.05)。结论 肝纤维化大鼠选择性门静脉栓塞、门静脉结扎后对侧肝脏均有再生能力;外源性HGF能明显提高肝再生能力。     

流式细胞术动态检测肝大部切除后再生相关因子的实验研究

为探讨肝细胞生长因子（HGF）、转化生长因子-α（TGF－α）、转子生长因子－β（TGF－β）在肝再生中的作用及机制。应用流式细胞术（FCM）动态检测再生肝细胞S期肝细胞百分含量及HGF、TGFα、TGF－β的变化。结果表明：HGF、TGF－α浓度在肝大部分切除6h后达高峰，而TGF－β24h后缓慢上升。提示HGF、TGF－α、TGF－β分别通过不同的机制影响肝再生，HGF、TGF－α为正性调节因子，TGF－β为负性调节因子。     

大鼠骨髓间充质干细胞分化为肝样细胞的实验研究

目的探讨大鼠骨髓间充质干细胞(MSCs)体外诱导分化为肝样细胞的能力。方法利用密度梯度离心法分离、纯化培养大鼠骨髓间充质干细胞,在特定的培养液中加入肝细胞生长因子(HGF)和表皮细胞生长因子(EGF)联合培养进行定向诱导,分别于1周和2周后通过逆转录-聚合酶链反应(RT-PCR)鉴定经诱导后细胞甲胎蛋白(AFP)和白蛋白(ALB)的表达。结果诱导1周后RT-PCR检测MSCs示AFP表达,诱导2周后的MSCs AFP呈阴性,而ALB检测结果呈阳性。结论MSCs在体外特定的条件下可定向诱导分化为肝样细胞。     

Value of D-Dimers in patients with acute aortic dissection

Objective: To evaluatel the value of D-dimers in patients with acute aortic dissection (AAD). Methods: This study consisted of 16 patients with AAD and 27 non-AAD patients. Serum D-dimets were measured by Sta-Liatest D-DI immunoturbidimetric assay. Results: D-dimer level was higher (P ＜ 0.001) in patients with AAD(7.91 ± 5.52 μg/ml) than that in non- AAD group(1.57±1.24 μg/ml). D-dimer was positive (＞0.4 μg/ml) in all patients with AAD and in 10 control group patients (37%). Among patients with acute AAD, D-dimers tended to be higher in Stanford A than in Stanford B (8.67 ± 4.31 μg/ml vs. 3.24±1.27 μg/ml, P ＜0.01). D-dimer values tended to be higher in more extended disease(3.84 ± 1.65 μg/ml, 8.57 ± 3.58 μg/ml and 11.87 ± 5.69 μg/ml in thoracic aorta, thoracic and abdominal aorta, thoracic and abdominal aorta and iliacal arteries, respectively, P ＜ 0.05 for both 8.57 ± 3.58 and 11.87 ± 5.69 vs. 3.84 ± 1.65 ). Including the control group into the analysis, we found a sensitivity of 100%, a negative predictive value of 100%, and a specificity of 66% and a positive predictive value of 64% for D-dimer in diagnosis of AAD in our patients with suspected AAD. Conclusion: D-dimer was elevated in patients with AAD. A negative D-dimer test result could be useful in excluding AAD.     

Research of combined liver-kidney transplantation model in rats

Objective： To set up a simple and reliable rat model of combined liver-kidney transplantation. Methods： SD rats served as both donors and recipients. 4℃ sodium lactate Ringer＇s was infused from portal veins to donated livers,and from abdominal aorta to donated kidneys, respectively. Anastomosis of the portal vein and the inferior vena cava （IVC） inferior to the right kidney between the graft and the recipient was performed by a double cuff method, then the superior hepatic vena cava with suture. A patch of donated renal artery was anastomosed to the recipient abdominal aorta. The urethra and bile duct were reconstructed with a simple inside bracket. Results： Among 65 cases of combined liver-kidney transplantation, the success rate in the late 40 cases was 77.5%. The function of the grafted liver and kidney remained normal. Conclusion： This rat model of combined liver-kidney transplantation can be established in common laboratory conditions with high success rate and meet the needs of renal transplantation experiment.     

BLOOD PRESSURE CHANGE WITH AGE IN SALT-SENSITIVE TEENAGERS

Objective To observe blood pressure change with age in salt-sensitive teenagers whose salt sensitivity were determined by repeated testing.Methods Salt sensitivity was determined through intravenous infusion of normal saline combined with volume-depletion by oral diuretic furosemide in 55 teenagers. After five years, salt sensitivity was re-examined and subject blood pressure was followed up. Blood pressure changes in salt-sensitive teenagers were compared to that of non-salt sensitive teenagers over five years.Results After 5 years, the repetition rate of salt sensitivity determined by intravenous saline loading is 92.7%. In teenagers with salt sensitivity on the baseline, both the systolic blood pressure increments and increment rates were much higher than non-salt sensitive teenagers (12.7±12.1 mmHg vs. 2.8±5.2 mmHg, P＜ 0.01; 12.2%± 12.0% vs. 2.5% ±4.4%, P＜ 0.001,respectively). There was a similar trend for diastolic blood pressure (8.4 ± 6.4 mmHg vs. 3.7 ± 6.4 mmHg, P = 0.052; 13.2% ±10.6 % vs. 6.8%± 10.1%, P = 0.053, respectively).Conclusions Salt sensitivity determined by intravenous saline loading showed good reproducibility. Blood pressure increments with age were much higher in salt-sensitive teenagers than non-salt sensitive teenagers, especially in terms of systolic blood pressure.     

安心颗粒对急诊经皮冠状动脉介入术后生活质量影响的研究

目的：评价使用安心颗粒对急诊经皮冠状动脉介入术（PPCI）术后生活质量的影响.方法：将160例接受PPCI的急性ST段抬高型心肌梗死患者随机分为安心颗粒组（术前顿服安心颗粒8.8g,术后安心颗粒4.4 g/次,每日2次）和对照组（仅接受基础药物治疗）.所有患者均服用阿司匹林、氯吡格雷和阿托伐他汀.分别在入院时、出院前1d、出院后180 d时,应用心肌梗死多维度量表（MIDAS）、中文版SF-36评价量表对患者生活质量评分.并观察术后30 d以内的出血并发症、血小板减少症发生情况.结果：入院时和出院前1d,两组患者的心肌梗死MIDAS、SF-36量表评分比较无差异（P＞0.05）;出院后180 d时,与对照组比较,安心颗粒组MIDAS、SF-36评分明显减低（P＜0.05）;组内与入院时比较,两组出院前1d、出院后180 d时,MIDAS、SF-36评分均降低（P＜0.05）.两组患者在随访期间均无大量出血、少量出血、重度和极重度血小板减少症发生,安心颗粒组有4例、对照组有7例发生不明显出血（P＞0.05）.两组发生轻度血小板减少症的患者数比较无差异（P＞0.05）.结论：PPCI使用安心颗粒,能改善急性ST段抬高型心肌梗死患者的生活质量,且不增加出血风险.     

The comparative studies of the influences of Urapidil and Nicardipine on sino-atrial node function, atrio-ventricular node function and hemodynamics

Objective:To investigate the influences of urapidil and nicardipine on rabbit sinus function,atrio-ventricular node function and hemodynamics.Methods:Thirty-two Angora's rabbits were selected and randomly divided into four groups.U1 group:urapidil 0.25 mg/kg;U2 group:urapidil 0.5 mg/kg;N1 group:nicardipine 10 μg/kg;N2 group:nicardipine 20 μg/kg.All these medicine were administrated within 30 seconds.Measurements were taken before and after the administration of urapidil or nicardipine for the following data:mean blood pressure(MAP),heart rate(HR),sino-atrial conduction time(SACT),maximal sinoatrial recovery time(SNRTmax)corrected sinus node recovery time(CSNRT),index of sinus node recovery time(SNRTI),Wenckebach A-V conduction frequency (WB),and P-R interval.Results:Significant MAP and HR changes were identified in all of the four groups before and after administration of both urapidil and nicardipine.No significant changes could be found in the rest of the parameters.Intergroup analysis showed that SACT and CSNRT of N1 and N2 groups were shorter than those of the U2 group(P＜0.01);the MAP decreased(P＜0.01)and the HR increased drastically(P＜0.01).Conclusions:Neither urapidil(0.25 mg/kg,0.5 mg/kg)nor nicardipine(10μg/kg,20μg/kg)has any significant influence on rabbit sinus function or rabbit atrio-ventricular node function.Nicardipine could be a better choice than urapidil for parafunctional sinus node patients.     

Expression of OPGmRNA and ODFmRNA in the patients of hip fracture due to osteoporosis

Objective：To investigate the gene expression of osteoprotegerin（OPG） and osteoclast differentiation factor（ODF） in the bone tissue of patients with hip fracture due to osteoporosis. Methods：OPGmRNA and ODFmRNA in the bone tissue in 50 cases of osteoporosis sufferers（over 50 years old） with hip fracture（Observer Group） and 30 cases of hip facture sufferers with no osteoporosis（Control group） were analyzed with the Semi-Quantitative RT-PCR method. Results：The mRNA expressed of ODF, OPG were both high in the patients with hip fracture. In the control group, the expression of OPG mRNA was observed, while the expression of ODF mRNA was very slight. Conclusion：Aged patients contained all signals including OPG, ODF that are essential for inducing osteoclastogenesis and promoting bone resorption.     

The clinicopathological and immuohistochemical analysis of solid-pseudopapillary tumor of the pancreas： report of 9 cases

Objective：To investigate the clinical features, pathological characteristics and immunophenotype of solid-pseudopapillary tumor of the pancreas（SPTP）. Methods：Nine surgically treated cases of SPTP were retrospectively reviewed. Hematoxylin and Eosin（HE） staining and immunohistochemical staining were used to analyze all cases, and the general clinical data was collected. Results：Six patients were asymptomatic except for a palpable mass. Two patients complained of vague-epigastric pain. One patient appeared jaundice. The tumor was encapsulated and solid tissues alternately with cystic tissues. Histologically, the histological structure of solid portion was pseudopapillary with a fibrovascular core. Tumor cells were uniform and medium-sized which were arranged in sheets ets or nests or pseudopapillary patterns. Immunohistochemical studies demonstrated that SPTP proved positive in vimentin（9/9 cases）, AAT（9/9 cases）, NSE（9/9 cases）, ACT（7/9 cases）, CK20（2/9 cases）, CgA（1/9 cases）, S-100（3/gcases）, PR（4/gcases）, Syn（3/9 cases） and CD56（5/9cases）, negative in CEA and ER. Conclusion：SPTP is a tumor predominantly occurring in young women frequently without special symptoms. This tumor has various characteristical histological patterns with different immunophenotype.     

Dynamic Changes in Serum Estradiol and Progesterone levels in Patients of Premenstrual Syndrome with Adverse Flow of Liver-qi

Objective:To probe into the influence of changes of ovarian hormones on the pathogenesis of the specific sub-type premenstrual syndrome(PMS)and reveal partial microcosmic mechanisms of adverse flow of liver-qi.Methods:Estradiol(E2)and progesterone(P)levels in serum were determined at different phases of menstrual cycle by radioimmunoassay.Results:In the group of PMS with adverse flow of liver-qi.the secretive peak value Of E2 and P at the follicular phase significantly decreased,and the secretive peak value at the luteal phase did not come into being.Conclusions:Low E2 and P secretive peak at the follicular phase and absence of secretive peak at the luteal phase is one of the microcosmic mechanisms of PMS with adverse flow of liver-qi.One of the pathophysiologic mechanisms of specific sub-type PMS is probably the continuous low level of E2and P.     

Real-time Three-dimensional Echocardiography in Assessment of Left Ventricular and Right Ventricular Volumes

Real-time three-dimensional echocardiography (RT3DE)is a new ultrasound technique that enables dynamic threedimensional visualization and quantification of the heart in real time. Investigation of feasibility and methodology of RT3DE in determining left ventricular (LV) and right ventricular (RV) volumes, RT3DE was performed in 35 normal adults using Philips SONOS 7500 system with a 2-4 MHz matrix array transducer. The 60°×60° "pyramid" volume database was obtained and analyzed on a TomTec echo workstation. Both LV and RV volumes were calculated with four 3DE methods (i.e. apical 2, 4, 8, and 16-plane) through manually tracing ventricular endocardial borders in end diastole and end systole. Stroke volumes were then calculated. LV volume was also measured by 2DE Simpson's rule using GE VIVID 7 ultrasound machine.     

SCREENING FOR A 21-CHROMOSOME ABNORMALITY IN PREIMPLANTED EMBRYOS OF ELDERLY WOMEN

Increasing maternal age is the only etiological factor unequivocally linked to Down's syndrome in humans. The occurrence rate of newborns with Down's syndrome is about 1/220 in women over 35 years old. However, the occurrence rate in embryos fertilized in vitro, of the elder woman is unclear. Using FISH we screened the number of chromosome 21 in preimplanted embryos of 5 elderly women (average age, 38.4 years) to study the feasibility and necessity of screening trisomy 21 in embryos in patients over 35 years old at the in vitro fertilization (IVF) center.