人VEGF受体Ⅱ基因3区的克隆、表达研究

用RT－PCR法从人胎儿脐静脉内皮细胞中克隆人VEGF受体Ⅱ基因3区，将其重组到pAYZ表达载体中，构建成人VEGF受体Ⅱ基因3区表达载体。将该载体转化大肝肠菌16C9，获得稳定表达，表达产物以可溶性状态存在；SDS－PAGE和Western blot分析显示，其分子量约为14kD，以纯化的表达产物免疫Balb/c小鼠，眼球采血制备抗血清，抗血清效价在10^3以上，并具有抑制VEGF诱导内皮细胞增殖的作用，在抑制肿瘤血管新生中具有潜在的应用前景。     

Ni－ZrO_2精细功能梯度材料的设计、制备和分析

基于我们以前发表的精细复合Ｎｉ－ＺｒＯ２的热渗流现象；采用薄层迭层法，设计和制备了２８种不同成分分布曲线Ｙ［ｐ，Ｙ（１）］（Ｘ）的Ｎｉ－ＺｒＯ２ＦＦＧＭ；并用电子探针和扫描电子声显微镜分析了其成分分布、应力分布和组织结构；最后得到一种成分分布曲线为Ｙ［１，４０］（Ｘ）的Ｎｉ－ＺｒＯ２ＦＦＧＭ，它具有较显著的应力缓和效果、较高的结构完整性和致密度．本文为Ｎｉ－ＺｒＯ２ＦＦＧＭ的优化设计、制备和研究其成分分布及应力分布提供了一种崭新而有效的方法．     

CSF－1R部分基因在大肠杆菌中表达、抗融合蛋白血清制备及丝氨酸／苏氨酸磷酸化的初步研究

鼠巨噬细胞集落刺激因子－１受体（ｍＣＳＦ－１Ｒ）部分序列与质粒ｐＧＥＸ－２Ｔ谷胱苷肽转移酶（ＧＳＴ）融合，融合蛋白ＧＳＴ－ＣＤ－Ｐｓｔ（胞浆区），ＧＳＴ－ＣＴｅｒｍ（Ｃ－末端）和ＧＳＴ－ＫＩ（激酶插入区）成功地在大肠杆菌ＪＭ１０９株表达。初步结果指出：（１）ＧＳＴ－融合蛋白在体外激酶分析中可以作为底物；（２）由ＰＫＡ导致的磷酸化可能具有生理学意义；（３）ｍＣＳＦ－１Ｒ被ＣＫＩＩ磷酸化。３２Ｐ标记ＧＳＴ－ＣＤ－Ｐｓｔ的磷酸氨基酸分析证实，ｍＣＳＦ－１Ｒ的胞浆区丝氨酸上被磷酸化，已制备抗ＧＳＴ－ＣＴｅｒｍ，ＧＳＴ－ＫＩ和ＧＳＴ－ＣＤ－Ｐｓｔ兔抗体。抗血清的筛选通过野生型３２Ｄ－ＣＳＦ－１Ｒ转染子免疫沉淀进行。     

纳米TiO2光诱导杀伤 Bel-7402人体肝癌细胞研究

在光诱导条件下，采用HE染色法和四甲基偶氮唑蓝比色法（MTT法），研究了纳米TiO2对Bel-7402人体肝癌细胞的杀伤作用，考察了在不同纳米TiO2浓度、不同光照时间下纳米TiO2的抑瘤效果，并且对抑瘤机制进行了探讨．结果发现：在光诱导条件下，适宜的TiO2浓度具有较高的抑瘤率，同时抑瘤过程表现出类似一级反应的动力学规律；在光诱导条件下，纳米TiO2产生的活性氧组分与癌细胞膜内外的生物大分子反应，引起广泛的细胞结构破坏；造成癌细胞内Ca^2＋离子稳态失去平衡；引发细胞微管相关蛋白2（MAP-2）表达的变化，促进微管发生重组，从而导致细胞凋亡和坏死．     

快速富立叶变换在爆破分析中的应用

美国矿务局开发成功一种计算机软件，应用快速富立叶变换方法（ＦＦＴ）估算双数据。这种软件可用来估算：１）地面振动时间变化数据的频率成分；（２）起爆器起爆时间的分散对地面振动频率谱的影响；３）照片来估算岩石爆堆的破碎分布。在完成１）、２）两项工作中使用的是一维软件，使用了Ｍｏｎｔｅ Ｃａｒｌｏ方法来模拟起爆器的分散。计算结果表示成计算机制的三维彩色轮廓图。为了采用照片数字图象技术，确定破碎块的大小分布     

腺病毒载体介导的人凝血因子IX基因的离体和活体表达

为血友病Ｂ的基因治疗探索了高效转移和表达载体，建立了重组腺病毒载体介导的基因转移系统（Ａｄｈｆｉｘ），并进行了离体和活体表达研究。结果显示：腺病毒离体基因转移效率可达９８％以上并可在多种细胞中高效表达，ｈＦＩＸ蛋白最高可达６．８６μｇ／１０６细胞／２４ｈｒ。小鼠血浆中ｈＦＩＸ蛋白含量最高可达１０７２ｎｇ／ｍｌ血浆，并可持续表达１０周左右；以腹腔注射效果最好；免疫抑制剂环磷酰胺可明显延长表达持续时间。结果表明重组腺病毒载体可介导ｈＦＩＸｃＤＮＡ在离体细胞和活体组织中高效转移和表达。     

离子强度对于牛精蛋白在大肠杆菌中的表达的影响

通过基因合成将大肠杆菌偏好的精氨酸密码子CGT取代野生型牛精蛋白基因中的稀有精氨酸密码子AGA和AGG，得到密码子优化的牛精蛋白基因，并将其克隆入原核表达载体pGEX-2T中，组装成表达载体pGEX-2T-PS。将这个表达载体转化入大肠杆菌表达菌株BL21中，经IPTG诱导，可得到一33kD融合蛋白表达带，首次成功地将牛精蛋白在大肠杆菌中进行了表达，但其表达量很低，约为总菌体蛋白的13％。增加表遂菌株培养液的离子强度，可将蛋白表达含量提高到28％，且蛋白表达量与离子强度呈正相关。纯化后的牛精蛋白可在体外条件下与DNA发生结合。     

Ni—ZrP2精细功能梯度材料的设计,制备和分析

基于我们以前发表的精细复合Ｎｉ－ＺｒＯ２的热渗流现象，采用薄层迭层法，设计和制备了２８种不同成分分布曲线的Ｎｉ－ＺｒＯ２ＦＦＧＭ；并用电子探针和扫描电子声显微镜分析了其成分布、应力分布和组织结构；最后得到一种成分分布为Ｙ「１．４０」（Ｘ）的Ｎｉ－ＺｒＯ２ＦＦＧＭ，它具有较显著的应力缓和效果，较高的结构完整性和致密度，本文为Ｎｉ－ＺｒＦＦＧＭ的优化设计制备和研究其成分发布及应力分布提供了一种崭新而有     

β-半乳糖苷酶基因(lacZ)在大型经济海藻裙带菜中的瞬间表达

用高压氦气式基因枪将SV40启动子驱动下的β-半乳糖苷酶基因（lacZ）导入大型经济海藻裙带菜不同部位的组织切块中，48小时后染色检测，在假根、孢子叶和叶片中均检测到了lacZ的瞬间表达，研究还发现，对于裙带菜组织块的基因枪法转化，压强1300psi优于1100psi。另外在裙带菜中没有发现半乳糖苷酶的染色本底。本文结果提示：基因枪法是有效的裙带菜遗传转化方法；lacZ可以作为裙带菜基因工程研究的报告基因，SV40启动子能有效驱动外源基因在裙带菜中表面，没有组织特异性。这是lacZ在裙带菜中表面的首次报道，结果可为裙带菜基因工程育种研究提供方法学基础。     

高分辨CT检查对肺部结节患者原位癌及浸润性癌鉴别诊断价值

目的：探究高分辨CT检查对肺部结节患者原位癌及浸润性癌的鉴别诊断价值。方法：将2021年4月至2022年4月苏州市第九人民医院收治的90例肺部结节患者作为研究对象，对所有患者采用高分辨CT检查，然后进行病理检查，分析相较于病理检查，高分辨CT检查原位癌及浸润性癌鉴别的诊断结果及诊断价值，包括准确率、阳性预测值和阴性预测值、诊断灵敏度、特异度以及AUC等；根据高分辨CT扫描结果，将所有患者分为原位癌组以及浸润性癌组，比较原位癌组以及浸润性癌组患者的结节病理特征。结果：病理检查中发现原位癌患者48例，浸润性癌患者42例；高分辨CT检查中发现原位癌患者41例，浸润性癌患者49例。高分辨CT检查诊断的灵敏度为87.78%(79/90)，阳性预测值为4.88%(2/41)，阴性预测值为18.37%(9/49)；高分辨CT诊断原位癌的AUC值为0.96,95%CI为0.90-0.99，灵敏度88.76%，特异度为91.43%；原位癌组与浸润性癌组密度、大小、混合性磨玻璃结节、血管牵拉征、边缘分叶差异均有统计学意义（均P<0.05），而两组部位、血管增粗及血管位置差异均无统计学意义（均P>...     

LHRH-functionalized superparamagnetic iron oxide nanoparticles for breast cancer targeting and contrast enhancement in MRI

This paper shows that superparamagnetic iron oxide nanoparticles (SPIONs) conjugated to luteinizing hormone releasing hormone (LHRH) (LHRH–SPIONs), can be used to target breast cancer cells. They also act as contrast enhancement agents during the magnetic resonance imaging of breast cancer xenografts. A combination of transmission electron microscopy (TEM) and spectrophotometric analysis was used in our experiments, to investigate the specific accumulation of the functionalized superparamagnetic iron oxide nanoparticles (SPIONs) in cancer cells. The contrast enhancement of conventional T2 images obtained from the tumor tissue and of breast cancer xenograft bearing mice is shown to be much greater than that in saline controls, when the tissues were injected with LHRH–SPIONs. Magnetic anisotropy multi-CRAZED images of tissues extracted from mice injected with SPIONs were also found to have enhanced MRI contrast in breast cancer xenografts and metastases in the lungs.     

Simultaneous analysis of glycosylated and sialylated prostate-specific antigen revealing differential distribution of glycosylated prostate-specific antigen isoforms in prostate cancer tissues

Aberrant protein glycosylation has been shown to be associated with disease progression and can be potentially useful as a biomarker if disease-specific glycosylation can be identified. However, high-throughput quantitative analysis of protein glycosylation derived from clinical specimens presents technical challenges due to the typically high complexity of biological samples. In this study, a mass spectrometry-based analytical method was developed to measure different glycosylated forms of glycoproteins from complex biological samples by coupling glycopeptide extraction strategy for specific glycosylation with selected reaction monitoring (SRM). Using this method, we monitored glycosylated and sialylated prostate-specific antigen (PSA) in prostate cancer and noncancer tissues. Results of this study demonstrated that the relative abundance of glycosylated PSA isoforms were not correlated with total PSA protein levels measured in the same prostate cancer tissue samples by clinical immunoassay. Furthermore, the sialylated PSA was differentially distributed in cancer and noncancer tissues. These data suggest that differently glycosylated isoforms of glycoproteins can be quantitatively analyzed and may provide unique information for clinically relevant studies.     

Fourier transform infrared attenuated total reflection analysis of human hair: comparison of hair from breast cancer patients with hair from healthy subjects

A comparative study of Fourier transform infrared attenuated total reflection (FTIR-ATR) spectra of 32 scalp and pubic hair samples from individuals diagnosed with breast cancer and those who were negative for breast cancer showed increases in the beta-sheet/disorder structures (relative to alpha-helix structures) and C-H lipid content of hair from breast cancer patients. Thus, the presence of breast cancer appears to alter the hair growth process, resulting in changes in the composition and conformation of cell membrane and matrix materials of hair fiber. These appear to be consistent with the changes observed in X-ray diffraction patterns for hair from breast cancer patients. A blind study of 12 additional hair samples using these FTIR-ATR spectral differences as markers correctly identified all four hair samples from cancer patients (100%). Two of these samples were from breast cancer patients. Of the remaining two samples analyzing positive for cancer, one was from a prostate cancer patient and one from a lung cancer patient. Thus, it appears that the mechanism that alters hair fiber synthesis in the three types of cancer may be similar. The blind study incorrectly identified as positive for cancer three hair samples from two apparently healthy individuals and one patient considered cured from prostate cancer.     

Epidermal growth factor-ferritin h-chain protein nanoparticles for tumor active targeting

Human ferritin H-chain protein (FTH1)-based nanoparticles possess a precisely assembled nanometer-scale structure and high safety. However, their applications for imaging and drug delivery towards cancer cells remain limited due to a lack of target specificity. Epidermal growth factor receptor (EGFR) is overexpressed in many malignant tissues including breast cancer, and has been used as a therapeutic target for cancer treatment. Herein, a genetic method is shown to generate EGF-FTH1 chimeric proteins. EGF-FTH1 nanoparticles with EGF on the surface are then produced. The data demonstrate that EGF-FTH1 nanoparticles, with a small size (11.8 ± 1.8 nm), narrow size distribution, and high biosafety, can specifically bind to and then be taken up by breast cancer MCF-7 cells and MDA-MB-231 cells, but not normal breast epithelial MCF-10A cells. In contrast, binding and absorption of nontargeted ferritin-based nanoparticles to breast cancer cells are negligible. In vivo studies show that EGF-FTH1 nanoparticles are accumulated in breast tumors in a mouse xenograft model. Interestingly, the concentration of EGF-FTH1 nanoparticles in the tumor site is significantly reduced when mice are pretreated with an excess of free EGF. These results imply that EGF-EGFR interaction plays an important role in regulating the tumor retention of EGF-FTH1 nanoparticles.     

The Fibrillar Matrix: Novel Avenues for Breast Cancer Detection and Treatment

Breast cancer is marked by large increases in the protein fibers around tumor cells. These fibers increase the mechanical stiffness of the tissue, which has long been used for tumor diagnosis by manual palpation. Recent research in bioengineering has led to the development of novel biomaterials that model the mechanical and architectural properties of the tumor microenvironment and can be used to understand how these cues regulate the growth and spread of breast cancer. Herein, we provide an overview of how the mechanical properties of breast tumor tissues differ from those of normal breast tissue and non-cancerous lesions. We also describe how biomaterial models make it possible to understand how the stiffness and viscosity of the extracellular environment regulate cell migration and breast cancer metastasis. We highlight the need for biomaterial models that allow independent analysis of the individual and different mechanical properties of the tumor microenvironment and that use cells derived from different regions within tumors. These models will guide the development of novel mechano-based therapies against breast cancer metastasis.     

Cocktail Strategy Based on Spatio‐Temporally Controlled Nano Device Improves Therapy of Breast Cancer

Metastatic breast cancer may be resistant to chemo‐immunotherapy due to the existence of cancer stem cells (CSC). Also, the control of particle size and drug release of a drug carrier for multidrug combination is a key issue influencing the therapy effect. Here, a cocktail strategy is reported, in which chemotherapy against both bulk tumor cells and CSC and immune checkpoint blockade therapy are intergraded into one drug delivery system. The chemotherapeutic agent paclitaxel (PTX), the anti‐CSC agent thioridazine (THZ), and the PD‐1/PD‐L1 inhibitor HY19991 (HY) are all incorporated into an enzyme/pH dual‐sensitive nanoparticle with a micelle–liposome double‐layer structure. The particle size shrinks when the nanoparticle transfers from circulation to tumor tissues, favoring both pharmacokinetics and cellular uptake, meanwhile achieving sequential drug release where needed. This nano device, named PM@THL, increases the intratumoral drug concentrations in mice and exhibits significant anticancer efficacy, with tumor inhibiting rate of 93.45% and lung metastasis suppression rate of 97.64%. It also reduces the proportion of CSC and enhances the T cells infiltration in tumor tissues, and thus prolongs the survival of mice. The cocktail therapy based on the spatio‐temporally controlled nano device will be a promising strategy for treating breast cancer.     

Imaging breast cancer cells and tissues using peptide-labeled fluorescent silica nanoparticles

The imaging of tumor cells and tumor tissue samples is very important for cancer detection and therapy. We have taken advantages of fluorescent silica nanoparticles (FSiNPs) coupled with a molecular recognition element that allows for effective in vitro and ex vivo imaging of tumor cells and tissues. In this study, we report on the targeting and imaging of MDA-MB-231 human breast cancer cells using arginine-glycine-aspartic acid (RGD) peptide-labeled FSiNPs. When linked with RGD peptide using the cyanogen bromide (CNBr) method, the FSiNPs exhibited high target binding to alphavbeta3 integrin receptor (ABIR)-positive MDA-MB-231 breast cancer cells in vitro. Further study regarding the ex vivo imaging of tumor tissue samples was also carried out by intravenously injecting RGD peptide-labeled FSiNPs into athymic nude mice bearing the MDA-MB-231 tumors. Tissue images demonstrated that the high integrin alphavbeta3 expression level of the MDA-MB-231 tumors was clearly visible due to the special targeting effects of the RGD peptide-labeled FSiNPs, and the tumor fluorescence reached maximum intensity at 1 h postinjection. Our results break new ground for using FSiNPs to optically image tumors, and may also broaden the applications of silica nanoparticles in biomedicine.     

Monitoring the progression of metastatic breast cancer on nanoporous silica chips

Breast cancer accounted for 15 per cent of total cancer deaths in female patients in 2010. Although significant progress has been made in treating early-stage breast cancer patients, there is still no effective therapy targeting late-stage metastatic breast cancers except for the conventional chemotherapy interventions. Until effective therapy for later-stage cancers emerges, the identification of biomarkers for the early detection of tumour metastasis continues to hold the key to successful management of breast cancer therapy. Our study concentrated on the low molecular weight (LMW) region of the serum protein and the information it contains for identifying biomarkers that could reflect the ongoing physiological state of all tissues. Owing to technical difficulties in harvesting LMW species, studying these proteins/peptides has been challenging until now. In our study, we have recently developed nanoporous chip-based technologies to separate small proteins/peptides from the large proteins in serum. We used nanoporous silica chips, with a highly periodic nanostructure and uniform pore size distribution, to isolate LMW proteins and peptides from the serum of nude mice with MDA-MB-231 human breast cancer lung metastasis. By matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and biostatistical analysis, we were able to identify protein signatures unique to different stages of cancer development. The approach and results reported in this study possess a significant potential for the discovery of proteomic biomarkers that may significantly enhance personalized medicine targeted at metastatic breast cancer.     

Detection of a thousand copies of miRNA without enrichment or modification

We report direct quantitative analysis of multiple miRNAs with a detection limit of 1000 copies without miRNA enrichment or modification. A 300-fold improvement over the previously published detection limit was achieved by combining capillary electrophoresis with confocal time-resolved fluorescence detection through an embedded capillary interface. The method was used to determine levels of three miRNA biomarkers of breast cancer (miRNA 21, 125b, 145) in a human breast cancer cell line (MCF-7). A 30 pL volume of the cell lysate with approximately a material content of a single cell was sampled for the analysis. MiRNA 21, which is up-regulated in breast cancer, was detected at a level of approximately 12 thousand copies per cells. MiRNAs 125b and 145, which are down-regulated in breast cancer, were below the 1000-copy detection limit. This sensitive method may facilitate the analysis of miRNA in fine-needle-biopsy samples and even in single cells without enrichment or modification of miRNA. Advantageously, the instrumental setup developed here can be reproduced by others as it requires no sophisticated custom-made parts.     

Detection and verification of glycosylation patterns of glycoproteins from clinical specimens using lectin microarrays and lectin-based immunosorbent assays

Aberrant glycosylation is a fundamental characteristic of progression of diseases such as cancer. Therefore, characterization of glycosylation patterns of proteins from disease tissues may identify changes specific to the disease development and improve diagnostic performance. Thus, analysis strategies with sufficient sensitivity for evaluation of glycosylation patterns in clinical specimens are needed. Here, we describe an analytical strategy for detection and verification of glycosylation patterns. It is based on a two-phase platform including a pattern discovery phase to identify the glycosylation changes using high-density lectin microarrays and a verification phase by developing lectin-based immunosorbent assays using the identified lectins. We evaluated the analytical performance of the platform using the glycoprotein standard and found that the lectin microarray could detect specific bindings of glycoprotein to lectins at the nanogram level and the lectin-based immunosorbent assay could be used for verification of protein glycosylation. We then applied the approach to the analysis of glycosylation patterns of two glycoproteins, which are highly expressed in prostate cancer in our prior studies, prostate specific antigen (PSA) and membrane metallo-endopeptidase (MME), from aggressive (AC) and nonaggressive prostate cancer (NAC) tissues. The observed differences in glycosylation patterns of PSA and MME may represent a significant clinical importance and could be used to develop multiplex assays for diagnosis of aggressive prostate cancer.