Regulation of mouse blastocyst adhesion,outgrowth and matrix metalloproteinase—2 by focal adhesion kinase

The interaction of extracellular matrix-integrin markedly influences the adhesion,outgrowth,differentiation and expression of serine proteinases by the blastocyst,so it is regarded as a vital factor in blastocyst implantation.Although the mechanism of extracellular interactions between extracellular matrix and integrins has been well elucidated,the roles of the signaling molecules in the extracellular matrix-integrin signal transduction pathway in blastocyst implantation are unknown.This limits the understanding of blastocyst implantation and ECM-integrin signal transduction pathway.In the present study,in vitro blastocyst culture and indirect immunocytochemistry,matrix metalloproteinases(MMPs) zymography and antisense oligodeoxynucleotide(ODN) were used to investigate the expression of a fundamental molecule of integrin-dependent signal transduction pathways,focal adhesion kinase(FAK),in mouse blastocysts and its influence on mouse blastocyst adhesion,outgrowth and MMP-2.The results showed that mouse blastocysts expressed FAK.FAK protein was clustered in the peripheral migrating trophoblast cells and dispersed in the central area of blastocyst outgrowth.Fibronectin triggered pro-MMP-2 and 64kD MMP-2 activities.The antisense ODN to FAK attnuated pro-MMP-2 and 64kD MMP-2 activites which decreased abruptly and tended to disappear with increasting concentrations of the antisense ODN.Both mouse blastocyst adhesion and outgrowth on fibronectin were also influenced by the antisense ODN.Up to 20μg/mL of the antisense ODN concentration,the adhesion and out-growth rates were decreased in a dose-dependent manner.The results indicated that FAK influenced mouse blastocyst adhesion,outgrowth and MMP-2 activity by intracellular signal transduction.In other words,FAK regulates mouse implantation in terms of blastocyst adhesive and invasive abilities.     

Effects of hyaluronic acid－chitosan－gelatin complex on the apoptosis and cell cycle of L929 cells

With the development in the field of tissue engineering, the interaction between biomaterials and cells has been deeply studied. Viewing the cells seeded on the surface of materials as an organic whole, cell cycle and apoptosis are analyzed to deepen the study of cell compatibility on biomaterials, while cell proliferation and differentiation are studied at the same time. In this paper, hyaluronic acid is incorporated into the chitosan-gelatin system. Propidium iodide (PI) was used in cell cycle analysis and the double-staining of cells with annexin-V and PI was applied in cell apoptosis analysis. The results show that incorporated hyaluronic acid shortens the adaptation period of cells on the material surface, and then cells enter the normal cell cycle quickly. In addition, added hyaluronic acid inhibits cell apoptosis triggered by the membranes. Therefore, hyaluronic acid improves the cell compatibility of chitosan-gelatin system and benefits the design of biomimetic materials.     

Effect of land use on microbial biomass－C, －N and －P in red soils

Eleven red soils varying in land use and fertility status were used to examine the effect of land useon microbial biomass -C, -N and -P. Microbial biomass-C in the red soils ranged from about 68 rag C/kg to 225 mg C/kg, which is generally lower than that reported from other types of soil, probably because of low or-ganic matter and high acidity in the red soils. Land use had considerable effects on the amounts of soil Cmic.The Cmic was the lowest in eroded fallow land, followed by woodland, tea garden, citrus grove and fallow grassland, and the highest in vegetable and paddy fields. There was significant correlation between Cmic and organic matter content, suggesting that the influence of land use on Cmie is mainly related to the input and ac-cumulation of organic matter. Microbial biomass-N in the soils ranged from 12.1 Nmg/kg to 31.7 Nmg/kg andwas also affected by land use. The change of Nmic with land use was similar to that of Cmic. The microbial C/N ratio ranged from 5.2 to 9.9 and averaged 7.6. The Nmic was significantly correlated with soil total N and available N. Microbial biomass-P in the soils ranged from 4.5 mg P/kg to 52.3 rag P/kg. The microbial C/P ratio was in the range of 4-23. The Pmic was relatively less affected by land use due to differences in fertili-zation practices for various land use systems.     

Distribution of a-MSH-like immunoreactive cells in the nervous system,Hatschek's pit and other tissues of amphioxus,Branchiostoma belcheri

Immunohistochemical localization of α-melanocyte-stimulating hormone （α-MSH） in the nervous system, Hatschek＇s pit and other tissues of amphioxus （Branchiostorna belcheri） was performed using the antibody against synthetic α-MSH. The results revealed that α-MSH-like immunoreactive cells were distributed at the dorsal side and ventral side of brain vesicle, the dorsal side and the surrounding of nerve tube, and in the epithelial cells of Hatschek＇s pit, the zone 1, 3, and 6 of endostyle and gut. The immunoreactive substance was also found in the primary oocytes of the small and large growth stage of ovary and early stage spermatogenic cells in testis. These findings indicate that α-MSH is an ancient and highly conserved hormone and it is extensively distributed in amphioxus. Although Hatschek＇s pit in amphioxus does not have a structure of the intermediate lobe of vertebrate adenohypophysis, it has already hosted α-MSH-like endocrine cells, implying that the functional differentiation of α-MSH-like cells occurred earlier than the differentiation of the tissue structure. The results of the present study provided a new evidence for the endocrinology of Hatschek＇s pit and for the origin and evolution of vertebrate adenohypophysis.     

A carbon－and oxygen－isotoperecord of recent environ－mental change from Qinghai Lake, NE Tibetan Plateau

A c. 300-year oxygen and carbon isotope record derived from fine-grained and ostracod carbonate from Qinghai Lake testifies to dramatic interannual to illterde-cadal linmologicai change. Fine-grained carbonates, whichare mainly authigenic, are likely to have formed in the epillmnion of the lake and their isotopic composition reflects the summer temperature and, more importantly, the isotopic composition of the near-surface waters, which is mainly afunction of evapor ative concentration. Ostracod shells aresecreted in the benthos of the lake, and their isotopic compo-sition reflects summer bottom-water conditions, together with fractionation effects, which may differ between species.Differences between contemporaneous values from authi-genic carbonates and ostracod shells may provide an indica-tion of stratification within the lake and variations in effec-tive precip itation over the northeast part of the Tibetan Pla-tcau over the past 300 years. A period of moderate evapora-tive concentration, from about 300 to 100 yr BP, was inter-rupted by a marked wet phase from-100 to 40 vr BP. which was in turn followed by a return to drier conditions in the most recent part of the record. The increase in δ^18O values in the latter part of the record accords well with instrumental records of lake-level lowering and salinity increase since about 1955 AD.     

JWA, a novel microtubule－associated protein, regulates homeostasis of intracellular amino acids in PC12 cells

Our previous studies demonstrated that JWA,a novel retinoic acids responsive and cytoskeleton related gene, is associated with cell differentiation and apoptosis. In the present study, to elucidate if the JWA is a novel kind of microtubnle-associated proteins (MAPs) and functiona llylink to microtubule, we first successfully identified JWA from the physically purified MAPs complex of rat braint issues. The results of co-immunoprecipitation, gene transfection and immunofluorescence microscopy assays from HBE and NIH3T3 cells provide strong evidence for a linkage between JWA and β-tubulin. In general, JWA is stably binding to β-tubulin whenever microtubule is polymerized or not,and it may be critical to the mitosis process. In addition, by use of the antisense oligonucleotides technique, we also showed that JWA is a negative modulator on intracellular amino acids in PC12 cells. Further analysis indicated that JWA selectively regulates both taurine, an inhibitory aminoacid, and glutamate, an excitatory amino acid. In conclusion,JWA is not only structurally associated, but also a novel functional MAP.     

Effect of Quercetin on Breeding and Apoptosis of Cervical Cancer HeLa Cell and on Growth of Transplanted Tumor in Nude Mice

Effect of quercetin on HeLa cell system of cervical cancer was studied by methods of MTT and Annexin V-FITC/PI. The results show that quercetin has functions of inhibiting breeding of HeLa cells and inducing apoptosis of the cells. The total apoptosis rate is positively proportional to reaction duration and concentration of quercetin used. The maximum apoptosis rate being （88.76±2.35）% was obtained when the concentration was 50.0 μmol/L and the cells were treated with quercetin for 72 hours. Based on establishing a model of tumor of cervical cancer transplanted into nude mice, quercetin of different concentrations was injected into abdominal cavity of nude mice and situation of tumor growth was reviewed. The result showed that with quercetin concent＇ration increasing from 0 to 100.0 μmol/L, the transplantation volume and weight of the tumors decreased from （279.59±70.58） mm^3 and （0.145±0.019） g to （128.72±36.12） mm^3 and （0.089± 0.019） g respectively, while apoptosis rate of the transplanted tumor increased from （9.63±1.85）% to （34,98±0.47）%, which proved that quercetin inhibited increment of volume and weight of transplanted tumor in nude mice bodies.     

MMP-2、MMP-9及其抑制剂在肺癌中的表达与临床意义

目的探讨肺癌组织中MMP2、MMP9及其抑制剂的表达与肺癌的生长、转移等临床特征及预后的关系.方法采用免疫组织化学法SP法检测45例肺癌组织中MMP2、MMP9及其抑制剂的表达,观察它们与肺癌的生长、TMN分期的关系.结果MMP2、MMP9、TIMP1和TIMP2在45例肺癌组织中的阳性表达率分别为57.78%(26/45)、37.78%(17/45)、35.56%(16/45)及51.11%(23/45);它们的表达与肺癌的原发灶范围、淋巴结转移、远处转移、TNM分期及平均生存时间有统计学意义(P<0.05).结论MMP2、MMP9及其抑制剂的表达与肺癌的TNM分期及平均生存时间关系密切,它们在组织中的高表达是判断肺癌预后的独立因素.     

CD44v6、E-cadherin和ki-67蛋白质与腮腺多形性腺瘤的生物学行为相关性研究

为检测腮腺多形性腺瘤中CD44v6、E-cadherin和ki-67蛋白表达,以探讨其与腮腺多形性腺瘤生物学行为的相关性。应用免疫组织化学SP法染色检测腮腺多形性腺瘤组、多形性腺瘤恶变组和瘤旁非瘤组织组各组间CD44v6、Ecadherin和ki-67蛋白表达。结果显示,(1)CD44v6、E-cadherin和ki-67在腮腺多形性腺瘤组中的阳性表达率分别为100%、96%和73.8%;(2)E-cadherin和ki-67在腮腺多形性腺瘤组、多形性腺瘤恶变组和瘤周非瘤组织间的阳性表达率均有差异,其表达与腮腺多形性腺瘤的浸润转移有关;(3)CD44v6在腮腺多形性腺瘤组与瘤周非瘤组织组,多形性腺瘤恶变组与瘤周非瘤组织组中的表达有统计学差异。由此可知,E-cadherin和ki-67与腮腺多形性腺瘤的生物学行为有关,可作为新的肿瘤标志物,其表达对判断预后有一定的价值。     

Expression and regulation of metalloproteinases-2, -9 and tissue inhibitors of metalloproteinases in rat corpus luteum

The expression and regulation of metalloproteinases-2, -9 (MMP-2, -9) and their tissue inhibitors TIMP-1, -2, -3 mRNA were studied in this experiment. In the PMSG- hCG primed pseudopregnant rat, MMP-2, -9 mRNA levels were the highest at Day 1, decreased from Day 4, and reached the minimal level at Day 8, then increased at Day 14; no significant changes were observed in TIMP-2 mRNA expression from Day 1 to Day 14; TIMP-3 mRNA expression was the lowest at Day 1, increased from Day 4, reached the maximal level at Day 8, and persisted to Day 14. TNF-αcould significantly increase the expression of MMP-2, -9 and TIMP-1 mRNA in the in vitro perfused pseudopregnant CL, and decrease the expression of TIMP-3 mRNA, but had no effect on TIMP-2 mRNA expression. The results indicate that MMP-2, -9 and TIMP-1, -2, -3 might be involved in the regulation of CL function and maintenance of CL structure via their coordinated gene expression. TNF-α could inhibit luteal regression via increasing MMP-2, -9 and TIMP-1 mRNA in the in vitro perfused pseudopregnant ovary.     

CD44v6和p16在宫颈上皮内瘤变及宫颈癌中的表达及意义

目的探讨CD44v6和p16表达产物在宫颈上皮瘤样病变及宫颈癌中表达的意义。方法对正常宫颈鳞状上皮、宫颈上皮内瘤变（CIN）和宫颈癌组织共95例,采用免疫组织化学SP法,对宫颈癌变过程中CD44v6和p16蛋白进行研究,将结果进行统计分析。结果CD44v6蛋白在CIN中的表述高于正常宫颈上皮（P〈0.01）,在宫颈癌中的表达也高于正常宫颈上皮（P〈0.01）,且高于CIN中的表达（P〈0.01）,p16蛋白在CIN中的表达高于正常宫颈上皮（P〈0.01）,在宫颈癌中的表达也高于正常宫颈上皮（P〈0.01）;CD44v6和p16两种蛋白在CIN和宫颈癌中的表达无明显差异。结论CD44v6和p16蛋白的表达与宫颈癌的发生有关,提示这两种蛋白有可能作为高危人群早期筛查的一种免疫组化指标。     

Advanced primary peritoneal carcinoma: clinicopathological and prognostic factor analyses

Objective： To investigate the factors favoring a positive prognosis for advanced primary peritoneal carcinoma （PPC）. Methods： Twenty-four cases meeting the criteria for PPC were analyzed retrospectively for the clinicopathologic profiles. Immunohistochemistry was used to determine the expressions of p53, Top2α, Ki-67 and Her-2/neu. Then all these clinicopathological factors and molecular markers were correlated with the prognosis. Results： There were 15 cases of primary peritoneal serous papillary carcinoma （PPSPC）, 6 cases of mixed epithelial carcinoma （MEC） and 3 cases of malignant mixed Mullerian tumor （MMMT）. All patients underwent cytoreductive surgery with optimal debulking achieved in 3 cases. Among those receiving first-line chemotherapy, 13 patients received the TP regimen （paclitaxel-cisplatin or carboplatin） and 7 patients received the PAC regimen （cisplatin-doxorubicin-cyclophosphamide）. The median overall survival of all patients was 42 months, while the breakdown for survival time for patients with PPSPC, MMT and MEC was 44, 13 and 19 months, respectively. The expressions of p53, Top2α and Ki-67 were all demonstrated in 11 cases respectively. None showed the expression of Her-2/neu. There were significant differences in the median survival between patients with PPSPC and those with MMMT （44 months vs 13 months, P〈0.05）, also between patients receiving TP combination and those receiving the PAC regimen （75 months vs 28 months, P〈0.05）. Another significant difference in the median progression-free survival （PFS） was identified between patients with positive p53 immunostaining and those with negative p53 immunostaining （15 months vs 47 months, P〈0.05）, whereas age, menopausal status, residual tumor size and the other molecular factors did not significantly impact survival. Conclusion： Patients with PPC should be treated with a comprehensive management plan including appropriate cytoreductive surgery and responsive chemotherapy. Overestimating an o     

胃印戒细胞癌中CD44v6，Survivin与PTEN的表达及临床病理学意义

探讨黏附相关因子变异体CD44v6、凋亡抑制基因Survivin和抑癌基因FIEN在胃印戒细胞癌组织中的表达与其临床病理特征之间的关系,及三者的关系。表明应用免疫组织化学SP法,检测CD44v6、Survivin和FIEN蛋白在73例胃印戒细胞癌组织中的表达。结果胃印戒细胞癌中CD44v6、Survivin和PIEN蛋白的阳性表达率分别为78．1％（57／73）、63．0％（46／73）、34．2％（25／73）。CD44v6随胃印戒细胞癌浸润深度加深、淋巴结的转移逐渐上升（P〈0．05）;随浸润深度加深、淋巴结的转移、器官转移、临床分期增加Survivin蛋白的阳性表达率逐渐上升（P〈0．01）,而FTEN基因的阳性表达率逐渐降低（P〈0．05）。在胃印戒细胞癌组织中CD44v6与PFEN蛋白的表达呈负相关（相关系数r=-0．214,P〈0．05）,Survivin与PTEN的表达呈负相关（r=-0．404,P〈0．01）,CD44v6与Survivin的表达呈正相关（r=0．28,P〈0．01）。因此可知,CD44v6、Survivin和PIEN基因的异常表达与胃印戒细胞癌临床病理特征有关,PIEN和Survivin基因表达可以作为判断胃印戒细胞癌生物学行为的指标;CD44v6与Survivin的协同作用,以及两者与PTEN拮抗作用共同参与胃印细胞癌的发生发展过程。     

三阴乳腺癌与基底细胞样乳腺癌病理形态和生物学特征探讨

目的探讨三阴乳腺癌(triple-negative breast cancer,TNBC)与基底细胞样乳腺癌(basal-likebreast cancer,BLBC)及非三阴乳腺癌(non triple-negative breast cancer,NTNBC)的关系及其形态、生物学特征。方法应用免疫组织化学(immunohistochemistry,IHC)方法对96例乳腺癌标本进行HER2、ER、PR蛋白的检查;应用FISH(fluorescent in situ hybridization)方法对3例HER2 IHC 3+及5例HER2 IHC 2+标本进行HER2基因扩增的检查;对22例TNBC进行了CK5/6或EGFR的检查。按照检查结果,将其分为TNBC、NTN-BC和BLBC,比较三者的病理形态及生物学特征。结果 NTNBC占72.92%(70/96),其组织学Ⅰ、Ⅱ及Ⅲ级的病例分别为28.57%(20/70)、57.14%(40/70)和14.29%(10/70),淋巴结转移率为44.44%(16/36);TN-BC占乳腺癌的27.08%(26/96),无组织学Ⅰ级病例,组织学Ⅱ及Ⅲ级的病例分别为53.85%,(14/26)及46.15%,(12/26)例,淋巴结转移率为61.11%(11/18);BLBC占TNBC的63.64%(14/22),同TNBC一样,无组织学Ⅰ级病例,Ⅱ、Ⅲ级的病例分别为57.14%(8/14)及42.86%(6/14),淋巴结转移率为55.56%(5/9)。TN-BC与BLBC低分化病例的比例以及淋巴结的转移率均高于NTNBC。3例HER2 IHC 3+的病例FISH检查结果同IHC,5例HER 2 IHC 2+标本FISH检查1例基因扩增,3例阴性,1例结果不确定。结论 TNBC与BLBC占乳腺癌1/4的比例,组织学多为中、低分化,浸润性生长,易发生淋巴结转移和复发,临床预后较差。虽然BLBC与TNBC大部分有重叠,但其具有独自的特异性,应成为独立的组织病理学类型。CK5/6或EGFR可用作从TNBC中筛选BLBC的指标。IHC 2+病例要做FISH检查,以正确指导治疗。     

组织芯片分析CD44v6和PCNA在口腔鳞癌及癌前病变中的表达及意义

目的：研究CDOv6和PCNA的表达与口腔鳞癌的发生、发展的关系。方法：应用组织芯片技术,通过免疫组织化学SP法分别检测一张组织芯片上117例组织样本中CD44v6、PCNA蛋白表达情况。结果：CD44v6在正常口腔黏膜组织和癌前病变呈阳性表达;癌组织中CD44v6的表达明显,但其表达程度随肿瘤恶性程度的增高而降低,PCNA的表达程度随着增生细胞和癌分级增加而升高。结论：在正常口腔黏膜上皮、增生上皮以及鳞癌中这两种蛋白的表达之间呈负相关。应用组织芯片大规模高效检测临床组织样本是可行的,具有快速、方便、经济、准确的特点。     

Expression and significance of cyclin D1, p27kip1 protein in bronchioloalveolar carcinoma

PURPOSE: To investigate the relationship between expression of cell cycle-related protein cyclin D1, p27kip1 and the pathogenesis of bronchioloalveolar carcinoma (BAC) and the value of prediction of prognosis. METHODS: Cyclin D1 and p27kip1 protein were detected by immunohistochemical En Vision method in 43 BACs. RESULTS: The positivity of cyclin D1 in BAC was 65.1% (28/43), which was significantly higher than that in normal pulmonary tissue (0/13), P<0.01. No statistically significant association was found between cyclin D1 expression data and sex, age, tobacco-use history, histologic subtype (mucinous vs nonmucinous), stromal fibrosis, lymph node metastasis, clinical stage or postoperative survival period (P>0.05), while cyclin D1 expression was found to be negatively correlated with tumor size (P<0.05). The positivity of p27kip1 in BACs was 51.2% (22/43), significantly lower than that in normal pulmonary tissue (12/13), P<0.01. p27kip1 expression level was not associated with sex, age, tobacco-use history, tumor size or histologic subtype (P>0.05), but was negatively correlated with stromal fibrosis, lymph node metastasis and clinical stage (P<0.05); and positively associated with postoperative survival period (P<0.01). The survival rate of p27kip1 positive group was significantly higher than that of p27kip1 negative group (P<0.01). No statistically significant correlation was found between cyclin D1 and p27kip1 expression. CONCLUSIONS: Increased cyclin D1 expression and decreased p27kip1 expression are related to the pathogenesis of BAC; decreased p27kip1 expression is associated with metastasis progression; immunodetection of p27kip1 is useful for assessment of prognosis.