目的 探讨蛇床子素(Ost)预处理对脂多糖(LPS)引起的急性肺损伤的防治作用并初步研究其作用机制.方法 BALB/C小鼠随机分为生理盐水对照组(NS组)、蛇床子素对照组(Ost组)、模型组(LPS组)及蛇床子素处理组(Ost+LPS组).气管滴注LPS建立小鼠急性肺损伤模型.造模6 h后测定肺湿/干质量比值(W/D)、肺泡灌洗液(BALF)中蛋白含量及肿瘤坏死因子(TNF-α)浓度,同时观察肺组织病理学改变.结果 LPS可明显提高小鼠肺W/D、BALF中蛋白含量和TNF-α浓度(P<0.05),肺组织结构发生明显的病理改变;Ost处理组可有效减轻LPS引起的上述变化(P<0.05),组织损伤程度明显减轻.结论 Ost预处理对LPS导致的小鼠急性肺损伤有保护作用,其保护机制可能与抑制LPS诱导的肺通透性增加有关.

术中快速冷冻切片病理诊断技术对手术中外科医师决定手术方案具有重要作用.但由于冷冻切片诊断时间仓促,取材局限,制片质量不如石蜡切片,冷冻切片延迟诊断或误诊的情况时有发生.故回顾分析我科术中快速冷冻切片诊断3303例,统计其与石蜡切片诊断的符合率,分析导致延迟诊断和误诊的原因,总结经验教训,进一步提高快速冷冻切片诊断的准确性.

The renin–angiotensin–aldosterone system (RAAS) plays an important role in the pathogenesis of acute lung injury (ALI)/acute respiratory distress syndrome (ARDS). Angiotensin converting enzyme 2 (ACE2) plays a protective role in acute lung injury. Osthole, a natural coumarin derivative extracted from traditional Chinese medicines, is known to have anti-inflammatory effect, but the effect of osthole on the ALI is largely unknown. The aim of this study is to explore whether and by what mechanisms osthole protects lipopolysaccharide(LPS)-induced acute lung injury. Herein, we found that osthole had a beneficial effect on LPS-induced ALI in mice. As revealed by survival study, pretreatment with high doses of osthole reduced the mortality of mice from ALI. Osthole pretreatment significantly improved LPS-induced lung pathological changes, reduced lung wet/dry weight ratios and total protein in BALF. Osthole also inhibited the release of inflammatory mediators TNF-α and IL-6. Meanwhile, osthole markedly prevented the loss of ACE2 and Ang1-7 in lung tissue of ALI mice. ACE2 inhibitor blocked the protective effect of osthole in NR 8383 cell lines. Taken together, our study showed that osthole improved survival rate and attenuated LPS-induced ALI and ACE2 may play a role in it.

术中快速冷冻切片病理诊断技术对手术中外科医师决定手术方案具有重要作用.但由于冷冻切片诊断时间仓促,取材局限,制片质量不如石蜡切片,冷冻切片延迟诊断或误诊的情况时有发生.故回顾分析我科术中快速冷冻切片诊断3303例,统计其与石蜡切片诊断的符合率,分析导致延迟诊断和误诊的原因,总结经验教训,进一步提高快速冷冻切片诊断的准确性。

目的 探讨乳腺尤因肉瘤/原始神经外胚层肿瘤(primitive neuroectodermal tumors,PNET)的病理诊断及鉴别诊断.方法 对1例乳腺尤因肉瘤/PNET进行免疫组化染色,并复习相关文献.结果镜下观察瘤组织由形态一致的小圆细胞所组成,呈实性片状或乳头状排列,局部可见菊形团样结构.免疫组化显示瘤细胞表达CD99、ER、CD56、GFAP、FLI-1、Calponin等.结论乳腺尤因肉瘤/PNET是少见的小圆细胞恶性肿瘤,CD99、FLI-1及神经内分泌标记物阳性对确诊有重要意义,ER可能在乳腺尤因肉瘤/PNET诊断中有一定的意义.

目的 探讨冷冻切片和石蜡切片诊断的符合率,分析冷冻切片误诊与延迟诊断的原因.方法 回顾性分析3 303例手术中快速冷冻送检标本的组织分布,与术后石蜡切片诊断符合率及误诊、延迟报告情况.结果 术中快速冷冻送检组织以甲状腺、乳腺、骨组织和卵巢居多,该组病例中确诊3 093例(93.64%),延迟诊断204例(6.18%),误诊6例(0.18%).造成延迟诊断和误诊的主要原因是取材局限、制片不良及对某些疾病的认识不足.结论 快速冷冻诊断是手术中明确病变性质的可靠手段,由于制片和诊断受诸多因素的制约,具有一定的误诊性.冷冻病理诊断要注意与临床医师的沟通,注意形态改变与临床表现相结合.技术操作需规范,保证制片质量.诊断中尤其加强对良、恶性病变的认识及鉴别能力,抓住病变特点,准确客观做出报告.对当时不能明确性质的病变要如实告知临床医师,可延迟诊断,避免误诊.定期复查总结,不断提高诊断符合率.

The cytotoxicity of a new nitroxyl nitroxide radical, tert-butyl-2 (4,5-dihydrogen-4,4,5,5-tetramethyl-3-O-1H-imidazole-3-cationic-1-oxyl-2-pyrrolidine-1-carboxylic ester (L-NNP) was examined in MCF-7 and MDA-MB-231 cells. L-NNP treatment resulted in a significant growth inhibition in MCF-7 and MDA-MB-231 cells. Compared with control, 10, 30, and 50 μg/ml L-NNP treatments for 48 h induced significant cell and nuclei swelling, and organelle distension. The marked cell death was seen in a concentration- and time-dependant manner in L-NNP treated groups. The L-NNP treated group displayed a concentration-dependant increase in DNA double strand damage compared to the control and the 1 Gy γ-rays exposure groups. These results suggest that L-NNP could result in more lethal genotoxicity than 1 Gy γ-radiation. Based on mitochondrial alteration (membrane potential loss and SDH activity descend), DNA damage, an increase in MDA production, and GSH-PX inactivation, we predicate that L-NNP induces lipid oxidation and oxidative damage in MCF-7 and MDA-MB-231 cells. Since L-NNP initiated a significant increase in reactive oxygen species, which could largely be inhibited by NAC pretreatment, the overall data strongly suggest that the mechanism of cytotoxicity of L-NNP was its ability to act as a strong free radical, and significantly increase intracellular reactive oxygen species production. This led to intracellular oxidative damage, and antioxidant enzyme inactivation, resulting in cell death. We hypothesize that the greater cytotoxicity of L-NNP in MDA-MB-231 cells than in MCF-7 cells might be due to more ROS production in MDA-MB-231 cells, leading to more oxidative damage.

To investigate the efiects of pulsed electromagnetic fleld (PEMF) with high electric fleld intensity on bone formation in murine osteoblast-like MC3T3-E1 cells, proliferation, alkaline phosphotase (ALP) activity, mineralized nodule formation, Collagen Type I (COL-I) and core-binding factor (Cbf)a1 mRNA expression, and bone morphogenetic protein (BMP)2/4 and mothers against decapentaplegic (Smad)1/5/8 protein expression were examined in cultured MC3T3-E1 cells after exposure to PEMF at the fleld intensities of 0kV/m, 50kV/m or 400kV/m for 400 consecutive pulses daily for 7 consecutive days. After 50kV/m of PEMF exposure, none of the above parameters of MC3T3-E1 cells changed signiflcantly when compared to the control groups. However, the proliferation, ALP activity and mineralized nodule formation of MC3T3-E1 cells in 400kV/m PEMF exposure groups decreased signiflcantly although COL-I and Cbfa1 mRNA expression and BMP2/4 and Smad1/5/8 protein expression did not change. The PEMF we used at high electric fleld intensity suppressed proliferation, difierentiation and

Objective:To investigate the PIK3CA gene mutation by MALDI-TOF mass spectrometry method(MassARRAY).Methods:Genomic DNA was extracted from paraffin embedding gastric cancer tissues.Targeted PIK3CA gene fragments were amplified by PCR.The remaining and nonincorporated dNTPs were removed from amplification products by Shrimp alkaline phosphatase(SAP).Mass spectrometry detection was carried out after primer extension reaction and primer extension reaction cleanup.Results:In 144 paraffin embedding gastric cancer tissues were detected,the mutation ratio of PIK3CA_E542K(1624G>A) was 77.6%,and the mutation ratio of PIK3CA_E545K(1633G>A) was 84%.All PIK3CA mutation results were the same as pyrosequencing results.Conclusion:The MAL-DI-TOF mass spectrometry method for PIK3CA mutation detection was successfully established.The mutation frequencies of PIK3CA_E542K(1624G>A) and PIK3CA_E545K(1633G>A) of gastric cancer have primarily been set up and this founding is only con-cerning with people of Han nationality among nor-thwest China.

OBJECTIVEWe examined alterations in the expression of tumorigenesis-related genes in the pituitary gland of rats exposed to electromagnetic pulses (EMP).METHODSThe global gene expression profiles of the pituitary gland in EMP-exposed and control groups were detected by cDNA microarray analysis. We then validated and further investigated the reduced expression of two tumorigenesis-related genes, Pten, and Jund, by assessing their mRNA and protein expression by quantitative real-time-PCR, western blotting, and immunohistochemistry in the pituitary gland of rats 6 months after exposure to EMP.RESULTSEMP exposure induced genome-wide gene expression changes in the rat pituitary gland. There was decreased expression of the Pten and Jund mRNAs and proteins in EMP-exposed rats compared with in unexposed control animals.CONCLUSIONEMP exposure alters the expression of tumorigenesis-related genes in the pituitary gland. These tumorigenesis-related genes are potentially involved in the development of pituitary gland tumors in rats.

Objective:To investigate the effect of chemotherapeutics on proliferation activity of tumor.Methods:Human cervical carcinoma Hela cells were divided into two groups:cells treated with or without DDP.MTT chromatometry method was used to evaluate the concentration of DDP for Hela cells.Immunochemistry method was used to detect the P27 expression.Cell flow cytometry was used to detect the change of cell cycle and apoptosis.IFFM-D chemoluminescence machine was used to detect the ultra weak luminescence intensity. Results:The IC50 of Hela cells treated with cis-DDP for 48h was 3mg/L.When cis-DDP concentration was lower than 3 mg/L, the cell toxicity showed dose-effect relation(P0.001).Cell flow cytometry results showed that Hela cells treated with DDP appeared more G2 phase and less G1 and S phase than control cell(P0.01);Compared with Hela treated with DDP,the apoptosis rates of Hela cells were increased signifcantly at 24 h,48 h and 72 h(11.4±5.8,21.8±7.9,32.5±11.6)%.P27 expression was increased in Hela cells treated with DDP.Luminous intensity of Hela cells treated with 10- 4 mol/L luminol and 0.3%H_2O_2 were significantly higher than that of Hela cells treated with DDP(P0.001).Conclusions:Ultra weak luminescence can reflect the metabolism and proliferation of tumor cells,which can be used for screening sensitive chemotherapy drugs.

目的:探讨人宫颈癌Hela细胞株经顺铂（DDP）作用后其超弱发光与细胞增殖活性变化的关系.方法:选择DDP诱导人宫颈癌Hela细胞株,比较人宫颈癌Hela,Hela+DDP细胞株形态变化.MTT比色法、流式细胞仪检测细胞生长周期变化及凋亡情况;用IFFM-D型流动式化学发光仪检测Hela,Hela+DDP细胞的超弱发光强度变化.结果:DDP对Hela细胞有生长抑制作用,并呈时间和浓度依赖性,其48 h的IC50值为3 mg/L,当DDP浓度在3 mg/L以下时,对Hela细胞无明显毒性作用,超过此浓度时,其毒性呈剂量效应关系（P＜0.01）.流式细胞仪结果表明,与Hela细胞相比较,Hela+DDP细胞G2期细胞数增多,而G1,S期的细胞数明显减少（P＜0.01）;细胞凋亡率在24,48,72 h分别为（11.4±5.8）%,（21.8±7.9）%,32.5±11.6)%.在1×10-4mol/L鲁米诺及3 mL/L双氧水（H2O2）条件下超弱发光强度随着时间的变化,Hela+DDP细胞明显降低（P＜0.01）.结论:在DDP非毒性剂量作用后,Hela+DDP细胞超弱发光强度降低,提示超弱发光检测可能作为筛选敏感化疗药物的一项指标.

任何生物在其生命活动的合成代谢过程中.都自发地辐射出一种极其微弱的光子流,称为生物超微弱发光,亦称超弱光子辐射(Ultraweak Plaoto Emission,简称UPE).有机体内的这种普遍发光现象与机体的氧化代谢、脱氧化作用、信息传递、光合作用,细胞的分裂和死亡以及生长调节作用都有密切关系[1].本次实验以大肠埃希菌为研究对象,榆测细菌加入抗生素后一段时间内UPE的数值变化,并结合药敏结果进行分析,现报告如下.

肝癌对化疗药物的敏感性低,其多药耐药是肝癌治疗失败的主要原因.P-糖蛋白(P gP)在肝癌组织中高表达,可能是参与肝癌多药耐药的关键分子[1],然而抑制P-gp的活性,并不能有效逆转肝癌的多药耐药[2].因此,除了P-gp,可能有其他的多药耐药分子共同参与了肝癌的多药耐药.乳腺癌耐药相关蛋白(BCRP)是近年来发现的一个新的多药耐药分子,可排泄多种化疗药物[3-4].已有研究结果显示,BCRP在肝癌中有表达,并可能是肝癌干细胞的一种标志物[5-6].然

Objective To study the variation of the intracellular oxygen free radicals after apoptosis induced by cisplatin in human cervical carcinoma cells.Methods Human cervical cancer cell line(Hela) were treated with different concentrations of DDP. The activation of the cell line,the changes in cell cycle,and the morphological changes of apoptosis cells were detected by MTT colorimetric assay,flow cytometry and electron microscopy,and the contents of intracellular malondialdehyde(MDA),superoxide dismutase (SOD) and glutathione(GSH) were detected by chemical colorimetric method.Results According to the MTT,the IC_(50) of DDP was 3 mg/L at the 48 h point,and DDP had no significant cytoxicity on Hela cell line when its concentration was less than 3 mg/L;contrarily, the effects were dose-dependent when the concentration of DDP was higher than 3 mg/L(P0.001).Compared with control group,the percentage of G2 phase cells increased(90.80±8.37)%in Hela+DDP group,while the number of G1 and S phase cells decreased significantly,the values were(30.84±8.02,5.30±0.60)%,and(P0.01),respectively.The typical morphological changes of the early apoptotic cells could be observed by scanning and transmission electron microscopy.Compared with the control group,the activities of SOD and GSH decreased significantly(P0.05) at the 48 hours point in the DDP group,and the concentration of MDA increased remarkably(P0.01).Conclusions DDP induces the death of Hela cells;the mechanism may be related to the oxidative damage in cells.

病理学作为一门桥梁学科在医学中起着举足轻重的作用,而实验课在病理学的教学中又占据着重要地位,引导学生上好病理实验课成为每位教员不可推卸的责任.为了上好实验课,对实习内容及实习方法进行了初步探索,取得了良好的效果.

This study was aimed to evaluate the reversed effects of cyclosporin A (CsA) on multidrug resistance (MDR) of human leukemic cell line HL-60/ADM, and to investigate the relationship of the oxygen free radical content between HL-60/ADM cells and the reversed HL-60/ADM cells (HL-60/ADM + CsA). The cytotoxicity and the reversed effects of CsA on multidrug resistance of human leukemic cell line HL-60/ADM were studied by MTT, flow cytometry (FCM) and immunohistochemical assay; the oxygen free radical for HL-60/ADM and HL-60/ADM + CsA cell lines were detected by colorimetric method. The results showed that the CsA less than 4 microg/ml had no significant cytotoxicity on HL-60/ADM, while the cytotoxicity was rised with CsA concentration increasing; And CsA (4 microg/ml) combined with ADM (1 microg/ml) could obviously restrain the growth of HL-60/ADM cells (p < 0.001). The P-gp expression of HL-60/ADM decreased obviously after exposure to CsA (4 microg/ml) for 72 hours, at the same cell conditions, MDA concentration of the reversed groups (HL-60/ADM + CsA cells) was higher than that of the control groups (HL-60/ADM cells) (p < 0.05), while the levels of SOD and GSH in the reversed groups were significantly lower than that in control groups (p < 0.001). It is concluded that MDR of HL-60/ADM can be reversed effectively by low dose of CsA, the level of oxygen free radical increases and the activity of antioxidants decreases in the reversed cells. Oxygen free radicals may be involved in this reverse process, which thereby lead to the cell death.

Objective To analyze the clinical,radiological and pathological characteristic of Telangiectatic Osteosarcoma. Methods The processing of diagnosis and treatment of the 2 cases with telangiectatic osteosarcoma was analyzed. Results Telangiectatic osteosarcoma is one kind of maligamant tumor that always happens in young people.Extensive osteolytic lesions could be found in X-ray films.Aneurismal bone cyst-like cavities with blood cells could be seen histologically. Heteromorphism and karyokinesis of oncocytes are histological characteristic for telangiectatic Osteosarcoma different from aneurismal bone cyst.Electrical microscope and immunohistological detection have much supplementary usage. Conclusion Clinical characteristic,radiological and histopathological detect of telangiectatic osteosarcoma,is the key for physicians from misdiagnosis.

Objective:To explore the diagnosing value of CT-guide pereutaneous pulmonary biopsy and testing the serum levels of tumor markers in diagnosis of lung carcinoma.Methods:A total of 335 cases of pulmonary nodules under- went atuo-spring-gun CT guide percutaneous pulmonary biopsy and analysis by pathology.Some of these cases were tested the serum levels of tumor markers.Results:All operations were successful with the sensitivity 100%.238 cases of malignant tumors were tested(the rate of diagnosis is 84.5%).Among them,there were 135 cases of adenocarcinoma,71 cases of squamous lung carcinoma,29 cases of small-cell lung carcinoma,19 cases of alveolar cell carcinoma,5 cases of mesothelioma and 25 cases of metastatic carcinoma.49 cases of benign lesion were tested.Among them,there were 39 cases of tuberculousis,10 cases were granuromatics and others,3 cases of undecided conclusion.After biopsy,27 cases were complicating hemoptysis(8.1%)and 22 cases of pneumatothorax(6.6%).The sensibility of CEA,NSE, CYFRA21-1 of diagnosing the lung carcinoma were 51.9% ,45.8%,and 62.8% respectively.Conclusion:CT-guide per- cutaneous pulmonary biopsy has a practical value for the diagnosis of lung cancer,particularly those with peripheral pul- monary nodules.The serum levels of CEA、NSE and CYFRA21-1 also play an important value to help the diagnosis.

To investigate the effects of electromagnetic pulse (EMP) on the morphology of the rat' s main organs (lung, kidney, liver, spleen, heart, testicle, ovary, and so on). Light microscope was used to observe the pathological changes in the main organs after the rats were exposed to EMP with respectively 1000, 1100 and 12000 pulses at different time point. After 24 hours exposure to 1000 and 1100 pulses of EMP, the alveolus epithelium were lightly swoln, dilatation and congestion were found in the blood vessel of the alveolar wall and the glomerulus. After 3 clays exposure, the morphostructure of alveolus, glomerulus and the renal tubule got right. There were no significant changes in other organs yet. Shortly after the 12000 pulses exposure and till 24 hours, congestion and swelling became more serious in the lung and the glomerulus with haemorrhage and eosinophils infiltration. The sacculus expanded evidently with a large amount of tubular protein casts. Congestions were also found in the liver, cardiac muscle, testicle, ovary, spleen and wall of digestive tract. The lesion aggravated gradually with dosages increasing. All these changes returned to normal after 3 days exposure. Results suggested that exposure to EMP of 12000 pulses would obviously increase the permeability of the blood vessel, especially in the organs with function of blood exchange barrier such as the glomerular filtration barrier or the air-blood barrier in alveolar wall. Thus the lung and the kidney are the sensitive organs to EMP exposure, their morphological and functional change is a responsive mechanism, which is recoverable.