手性金属配合物d,l-[Ni(phen)_3]^(2+)经Caco-2细胞单层模型的吸收转运

目的:研究手性金属配合物经Caco-2细胞单层转运吸收的手性选择性。方法:采用动力学研究方法,测定手性金属配合物跨Caco-2细胞单层的转运速率。结果:金属配合物的手性不同,导致金属配合物经Caco-2细胞单层的转运速率不同。结论:手性金属配合物经Caco-2细胞单层的转运吸收存在手性选择性的差异。

自体造血干细胞移植(ASCT)配合中药治疗多发性骨髓瘤1例

目的:评价自体外周血造血干细胞移植配合中药治疗恶性血液病MM的临床疗效。方法:1例确诊多发性骨髓瘤患者,接受自体外周血造血干细胞移植配合中药治疗。结果:移植后造血重建快,移植相关并发症减少,改善MM预后的重要手段。结论:其是治疗、根治MM的重要手段。

关节镜下髓芯减压自体干细胞体外培养回植术的手术配合

目的探索运用关节镜的微创技术有效清除死骨及准确植入骨髓间充质干细胞治疗早期股骨头缺血性坏死的手术配合。方法通过关节镜监视下髓芯减压将自体干细胞体外培养回植。结果所有随访病人根据百分法评价股骨头缺血性坏死疗效，优8例，良4例，可1例，优良率92．3％，所有切口均Ⅰ期愈合。结论此手术要求护士具备严格的无菌观念、熟练的技术操作和扎实的理论基础，通过与手术医生、麻醉师的密切配合保证手术的顺利进行。

上皮样滋养细胞肿瘤的手术配合

上皮样滋养细胞肿瘤（epithelioidtrophoblastictumor，ETT）是妊娠滋养细胞肿瘤（gestationaltrophoblasticneoplasms，GTN）中罕见的一种类型．通常继发于产后。其发病机制至今尚未明确，治疗前确诊非常困难。我院曾收治2例ETr患者，术前制定了周密的手术方案和应急措施，经充分的术前评估、术前准备和有效的术中护理配合。结合术后化疗，均取得了良好的治疗效果，现将手术配合的护理体会介绍如下。

液基细胞学配合阴道镜检查在诊断宫颈病变中的临床应月及意义

目的:探讨液基细胞学(TCT)配合阴道镜检查对宫颈病变的诊断价值。方法:对3010例来我院就诊的妇科患者行液基细胞学检查,对阳性涂片360例及可疑病例33例行阴道镜下活组织检查。结果:3010例涂片中阳性360例(11.96%)。其中ASC 253例(8.4%);LSIL 74例(2.46%);HSIL 27例(0.9%);SCC 6例(0.2%)。细胞学检查与阴道镜下活检病理结果的符合率分别为LSIL64.35%(74/115)、HSIL93.19%(27/29)、SCC100%(6/6)。结论:采用TCT联合阴道镜下活检能提高宫颈病变的阳性的诊断率。

红细胞抗体的持续存在性

由于输血、妊娠、组织移植使个体暴露于外来抗原而产生同种抗体,为了检测受血者血清中具有临床意义的同种抗体,输注前须进行红细胞配合性试验.对于不同人和不同抗体而言,抗体持续存在性的血清检测水平也是不同的.抗体免疫球蛋白的类型、数量及筛选方法的敏感性是影响输血前检测的主要因素.抗体一旦被检出,病人和医生应意识到必须避免不配合输血(特别是当病人在不同医院治疗时,医院应出具这一结果),选择缺乏相应抗原的血液输注.对于曾经检出的具有临床意义的同种抗体,即便现在检测是阴性,也应寻找配合性输血以避免迟发性溶血性输血反应(DHTR)的发生.人们至今还未统计出DHTR的发生率和死亡率,这与诊断标准和所涉及的抗体有关.DHTR常常因其反应的温和性和自限性而被忽略,然而也有一些DHTR导致贫血需要输血.本文回顾调查了有临床意义的红细胞同种抗体经历20年周期的持续存在性.

ABO、RhD配合型输注与同型输注红细胞的回顾性病例对照研究

目的:通过回顾性病例对照研究探讨ABO及RhD的配合型输注与同型输注红细胞的临床输血效果以及安全性评价,为临床输血方案选择提供依据。方法:随机选取2017-01-01—2019-06-30在山东大学齐鲁医院进行过ABO及RhD配合型输注红细胞的患者为病例组,同期进行ABO及RhD同型输注红细胞的患者为对照组,根据病例组患者的年龄、性别、红细胞输血量、输血时间以及所患疾病等特点匹配对照组。通过本院输血管理系统查询输注前及输注后24 h、48 h的Hb、RBC、Hct,采用重复测量方差方法分析病例组与对照组之间2种不同输血方案(ABO及RhD的配合型输注红细胞与同型输注红细胞)对患者的Hb、RBC、Hct影响有无显著差别以及输注红细胞前后2组患者的Hb、RBC、Hct有无显著差别,并分析这2种因素之间是否存在交互作用。通过分析病例组与对照组在输注红细胞前及输注后1 d、3 d、7 d、15 d的间接胆红素(IBiL)、因输注红细胞而新产生不规则抗体、新产生DAT阳性、输血不良反应等指标评价其临床输注安全性。结果:病例组与对照组在年龄、性别、红细胞输血量以及疾病分布等方面均差异无统计学意义(P>0.05);病例组与对照组的输注红细胞有效性差异无统计学意义(P>0.05);与输注红细胞前相比,病例组与对照组在输注红细胞后(24h及48h)的Hb、RBC、Hct与各自输注前相比均差异有统计学意义(P<0.01),各自输注红细胞后24 h与48 h相比均差异无统计学意义(P>0.05);病例组与对照组的2种不同输注红细胞方案对患者的Hb、RBC、Hct影响差异无统计学意义(P>0.05);红细胞不同输注方案与输注前后这2种因素之间不存在交互作用(P>0.05),即输注前与输注后的Hb、RBC、Hct均不随患者进行ABO及RhD的配合型输注红细胞或同型输注红细胞变化而变化。2组患者的IBiL在输注红细胞前及输注后1 d、3 d、7 d、15 d均差异无统计学意义(P>0.05),与对照组比较差异无统计学意义(P>0.05)。无因本次输注红细胞而产生新不规则抗体、新产生DAT阳性及出现不良输血反应的病例。结论:ABO及RhD配合型输注与同型输注红细胞同样安全有效,配合型输注红细胞在抢救某些危重患者生命及某些疾病治疗方面具有重要临床意义。

Desensitization of T lymphocyte function by CXCR3 ligands in human hepatocellular carcinoma

AIM: Despite the presence of lymphocyte infiltration, human hepatocellular carcinoma (HCC) is typically a rapidly progressive disease. The mechanism of regulation of lymphocyte migration is poorly understood. In this study,we investigated various factors regulating T cell migration in HCC patients. We examined serum CXC chemokine levels in HCC patients and demonstrated the production of CXC chemokines by HCC cell lines. We determined the effect of both HCC patient serum and tumor cell conditioned supernatant upon lymphocyte expression of chemokine receptor CXCR3 as well as lymphocyte migration. Lastly,we examined the chemotactic responses of lymphocytes derived from HCC patients.METHODS: The serum chemokines IP-10 (CXCL10) and Mig (CXCL9) levels were measured by cytometric bead array (CBA) and the tumor tissue IP-10 concentration was measured by ELISA. The surface expression of CXCR3 on lymphocytes was determined by flow cytometry. The migratory function of lymphocytes to the corresponding chemokines was assessed using an in vitro chemotactic assay. Phosphorylation of extracellular signal-regulated kinase (ERK) was determined by Western blot analysis.RESULTS: Increased levels of IP-10 and Mig were detected in HCC patient serum and culture supernatants of HCC cell lines. The IP-10 concentration in the tumor was significantly higher than that in the non-involved adjacent liver tissues.HCC cell lines secreted functional chemokines that induced a CXCR3-specific chemotactic response of lymphocytes.Furthermore, tumor-cell-derived chemokines induced initial rapid phosphorylation of lymphocyte ERK followed by later inhibition of ERK phosphorylation. The culture of normal lymphocytes with HCC cell line supernatants or medium containing serum from HCC patients resulted in a significant reduction in the proportion of lymphocytes exhibiting surface expression of CXCR3. The reduction in T cell expression of CXCR3 resulted in reduced migration toward the ligand IP-10, and both CD4+ and CD8+ T cells from HCC patients exhibited diminished chemotactic responses to IP-10 in vitro compared to T cells from healthy control subjects.CONCLUSION: This study demonstrates functional desensitization of the chemokine receptor CXCR3 in lymphocytes from HCC patients by CXCR3 ligands secreted by tumor cells. This may cause lymphocyte dysfunction and subsequently impaired immune defense against the tumor.

Fiber-modified adenoviral vector expressing the tumor necrosis factor-related apoptosis-inducing ligand gene from the human telomerase reverse transcriptase promoter induces apoptosis in human hepatocellular carcinoma cells

AIM: Because of a major resistance to chemotherapy, prognosis of hepatocellular carcinoma (HCC) is still poor. New treatments are required and gene therapy may be an option. Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) induces apoptosis in multiple malignant tumors, and using adenoviral vectors has shown a targeted tumor-specific therapy. However, repeated administration of adenoviral vectors can lead to cell resistance, which may be caused by the initial coxsackie-adenovirus receptor (CAR). One technique to overcome resistance is the use of modified adenoviral vectors containing an Arg-Gly-Asp (RGD) sequence. In this study we constructed an adenoviral vector (designated Ad/TRAIL-F/RGD) with RGD-modified fibers, expressing the TRAIL gene from the human telomerase reverse transcriptase (hTERT) promoter, and evaluated its antitumor activity in HCC cell lines.METHODS: To investigate the effects of Ad/TRAIL-F/RGD in human HCC cell lines Hep G2 and Hep 3b, cells were infected with Ad/CMV-GFP (vector control), Ad/gTRAIL (positive control), and Ad/TRAIL-F/RGD. Phosphatebuffered saline (PBS) was used as control. Cell viability was determined by proliferation assay (XTT), and apoptosis induction by fluorescence activated cell sorting (FACS).RESULTS: Cells treated with Ad/TRAIL-F/RGD and Ad/ gTRAIL showed a significantly reduced cell viability in comparison to PBS and Ad/CMV-GFP treatment in both cell lines. Whereas, treatment with PBS and Ad/CMVGFP had no cell-killing effect. The reduced cell viability was caused by induction of apoptosis as shown by FACS analysis. The amount of apoptotic cells was similar after incubation with Ad/gTRAIL and Ad/TRAIL-F/RGD. CONCLUSION: The new RGD modified vector Ad/TRAILF/RGD could become a potent therapeutic agent for the treatment of HCC, adenovirus resistant tumors, and CAR low or negative cancer cells.

Effect of ligand of peroxisome proliferator-activated receptor γ on the biological characters of hepatic stellate cells

AIM： To study the effect of rosiglitazone, which is a ligand of peroxisome proliferator-activated receptor gamma （PPARy）, on the expression of PPARy in hepatic stellate cells （HSCs） and on the biological characteristics of HSCs. METHODS： The activated HSCs were divided into three groups： control group, 3 μmol/L rosiglitazone group, and 10 μmol/L rosiglitazone group. The expression of PPARγ, α-smooth muscle actin （α-SMA）, and type Ⅰ and Ⅲ collagen was detected by RT-PCR, Western blot and immunocytochemical staining, respectively. Cell proliferation was determined with methylthiazolyltetrazolium （MTT） colodmetric assay. Cell apoptosis was demonstrated with flow cytometry. RESULTS： The expression of PPARγ at mRNA and protein level markedly increased in HSCs of 10 μmol/L rosiglitazone group （tvalue was 10.870 and 4.627 respectively, P〈0.01 in both）. The proliferation of HSCs in 10 μmol/L rosiglitazone group decreased significantly （t = 5.542, P〈0.01）, α-SMA expression level and type Ⅰ collagen synthesis ability were also reduced VS controls （tvalue = 10.256 and 14.627 respectively, P〈0.01 in both）. The apoptotic rate of HSCs significantly increased in 10 μmol/L rosiglitazone group vs control （X^2= 16.682, P〈0.01）. CONCLUSION： By increasing expression of PPARγ in activated HSCs, rosiglitazone, an agonist of PPARγ, decreases α-SNA expression and type Ⅰ collagen synthesis, inhibits cell proliferation, and induces cell apoptosis.

Functional expression of a proliferation-related ligand in hepatocellular carcinoma and its implications for neovascularization

AIM： To detect the expression of a proliferation-related ligand on human hepatocellular carcinoma （HCC） cell lines （SK-Hepl, HLE and HepG2） and in culture medium. METHODS： APRIL expression was analyzed by Western blotting in HCC cell lines. Effects of APRIL to cell count and angiogenesis were analyzed, too. RESULTS： Recombinant human APRIL （rhAPRIL） increased cell viability of HepG2 cells and, in HUVEC, rhAPRIL provided slight tolerance to cell death from serum starvation. Soluble APRIL （sAPRIL） from HLE cells increased after serum starvation, but did not change in SK-Hepl or HepG2 cells. These cells showed down-regulation of VEGF after incubation with anti-APRIL antibody. Furthermore, culture medium from the HCC cells treated with anti-APRIL antibody treatment inhibited tube formation of HUVECs. CONCLUSION： Functional expression of APRIL might contribute to neovascularization via an upregulation of VEGF in HCC.

Transfection of apoptosis related gene Fas ligand in human hepatocellular carcinoma cells and its significance in apoptosis

AIM: To evaluate the expression of apoptosis related gene Fas ligand (FasL) in human hepatocellular carcinoma (HCC) cells HepG2 and its significance in apoptosis. METHODS: Levels of soluble Fas ligand (sFasL) in a group of patients with hepatitis B virus (HBV)-induced chronichepatitis, HBV-positive liver cirrhosis and HCC were evaluated. In a further study, the recombinant eukaryotic expression plasmid pcDNA3.1hisB-FasL was transfected into HCC cells HepG2 by lipofection, and then soluble FasL was examined in the supernatant of culture cells by EIA, FasL expression in HepG2 cells was detected by immuohistochemistry. After being stained by annexin V and propidium iodine, cells were passed through a flow cytometer and examined by a fluorescence microscope and a laser scanning microscope.RESULTS: The sFasL levels were significantly lower in patients with HCC when compared to the patients with hepatitis or liver cirrhosis. In comparison with untransfected cells, the soluble FasL could be detected in the supernatant of transfected cells. FasL was expressed on the membranes and cytoplasm of transfected cells. The apoptotic cell rate was 36.30% in transfected cells, and was 11.53% in untransfected cells. Moreover, the different stage of apoptotic cells could be distinguished by annexin V and propidium iodine staining.CONCLUSION: Fas ligand is an apoptotic pathway of HCC cells.

Role of CD97^(stalk) and CD55 as molecular markers for prognosis and therapy of gastric carcinoma patients

Objectives： To explore the mechanism of development and aggressiveness in gastric carcinomas by investigating the expression and role of CD97 and its cellular ligand CD55 in gastric carcinomas. Methods： Tumor and corresponding normal mucosal tissue, collected from 39 gastric carcinoma patients, were examined by immunohistochemistry and RT-PCR for the expression of CD97 and CD55. Results： CD97^stalk was strongly stained on scattered tumor cells or small tumor cell clusters at the invasion front of gastric carcinomas. The expression of CD97^stalk was frequently observed in tumors of stage Ⅰ and T1 gastric carcinoma patients. The expression of CD97^stalk between Stage Ⅰ and Stage Ⅱ, Ⅲ, Ⅳ specimens showed significant difference （P〈0.05）, between T1 and T2, T3, T4 specimens also showed significant difference （P〈0.05）. Specimens with tumor invasion depth limited in mucosa of T1 specimens showed higher positive CD55 expression than specimens with the same tumor invasion depth in T2, T3, T4 specimens, the expression of CD55 between T1 and T2, T3, T4 specimens was significantly different （P〈0.05）. There was strong correlation between the distribution patterns of CD97^stalk and CD55 on tumor tissues （r=0.73, P〈0.05）. Signet ring cell carcinomas frequently contained strong CD97^stalk and CD55-staining. Conclusions： Our results suggest that CD97^stalk is probably involved in the growth, invasion and aggressiveness of gastric carcinomas by binding its cellular ligand CD55. CD97^stalk and CD55 could be useful as molecular markers for prognosis and therapy of gastric carcinoma patients.

Analysis of Heterosis, Combining Ability and Heritability of Cadmium Content in Brown Rice of Three-line Indica Hybrid Rice

Five cytoplasmic male sterile （CMS） lines were used as parents in an incomplete diallet cross and six restorer lines of rice design （Nc II design）. Thirty hybrid combinations with the same growth period were selected as experimental ma- terials to investigate the heterosis, combined ability and heredity of Cd content in brown rice of indica hybrid rice. According to the results, Cd content in brown rice showed a significantly negative heterosis; the general combining ability and specific combination ability of Cd content in CMS and restorer lines both reached extremely significant level （P〈0.01）, indicating that both genetic improvement of parents and e- valuation of combinations are important to the breeding of hybrid combinations with low accumulation of Cd; the broad-sense heritabitity and narrow-sense heritability of Cd content were both relatively high with slight differences, which respectively reached 97.73% and 80.10%, indicating that Cd content in brown rice mainly de- pends on the additive action of genes; in addition, parent improvement showed bet- ter effect on the selection of early generation.

Mechanism of counterattack of colorectal cancer cell by Fas/Fas ligand system

AIM： To determine the role of Fas/Fas ligand （FasL） in the immune escape of colon cancer cells. METHODS： Immunohistochemistry was used to observe the expression of Fas and FasL in the tissues of colon cancer patients. In situ hybridization was used to detect the localization of FasL mRNA expression in cancer tissues. Terminal deoxynucleotide transferase-mediated dUTP nick end labeling （TUNEL） assay and CD45 staining were performed to detect the apoptosis of tumor-infiltrating lymphocytes （TILs）. Co-culture assays of colon cancer cells （SW480） and Jurkat cells （Fas-sensitive cells） were performed to observe the counterattack of colon cancer cells to lymphocytes. RESULTS： Of 53 cases of colon carcinomas, 23 cases （43.4%） expressed Fas which was significantly lower as compared to the normal colonic mucosa （73.3%, P〈0.01）, and 45 cases （84.9%） of colon carcinomas expressed FasL, whereas only two cases （3.75%） in normal mucosa expressed FasL. FasL expression in the colon cancer cells was found to be associated with increased cell death of TIEs. The apoptotic rate of TIL in the FasL-positive staining regions of tumor cells was significantly higher than that in the FasL-negative staining region （54.84±2.79% vs 25.73±1.98%, P〈0.01）. The co-culture of SW480 cells and Jurkat cells confirmed the function of FasL on the SW480 cells. The apoptotic rates of Jurkat cells were found to be related with the amount of SW480 cells. CONCLUSION： Colon cancer cells can escape the immune surveillance and killing via decreasing Fas expression, and can counterattack the immune system via increasing FasL expression. Fas/FasL can serve as potential targets for effective antitumor therapy.

Construction of recombinant eukaryotic expression plasmid containing murine CD40 ligand gene and its expression in H22 cells

AIM: To construct a recombinant murine CD40 ligand (mCD40L) eukaryotic expression vector for gene therapy and target therapy of hepatocellular carcinoma (HCC).METHODS: mCD40L cDNA was synthesized by RT-PCR with the specific primers and directly cloned into T vector to generate middle recombinant. After digestion with restriction endonuclease, the target fragment was subcloned into the multi-clone sites of the eukaryotic vector. The constructed vector was verified by enzyme digestion and sequencing,and the product expressed was detected by RT-PCR and immunofluorescence methods.RESULTS: The full-length mCD40L-cDNA was successfully cloned into the eukaryotic vector through electrophoresis,and mCD40L gene was integrated into the genome of infected H22 cells by RT-PCR. Murine CD40L antigen molecule was observed in the plasma of mCD40L-H22 by indirect immuno-fluorescence staining.CONCLUSION: The recombined mCD40L eukaryotic expression vector can be expressed in H22 cell line. It providesexperimental data for gene therapy and target therapy ofhepatocellular carcinoma.

Social-Aware Joint Mode Selection and Link Allocation for Device-to-Device Communication Underlaying Cellular Networks

Joint mode selection and link allocation are crucial to achieve the advantage of Device-to-Device(D2 D) communications in improving spectral efficiency. In practice, cellular users tend to not be totally altruistic or absolutely selfish. How to stimulate them to devote their links and how to allocate their links to D2 D pair candidates efficiently are two main challenges. In this paper, we encourage cellular users through the variable payment with regard to the social tie strength between cellular users and D2 D pair candidates. In particular, the social tie strength is inferred through a graph inference model and its impact on the payment is quantified as a negative exponential function. Then, we propose a resource scheduling optimization model based on the non-transferable utility coalition formation game, and a distributed coalition formation algorithm based on the Pareto preference and merge-and-split rule. From them, the final coalition structure is obtained, which reflects the strategy of mode selection and link allocation. Numerical results are presented to verify the effectiveness of our proposed scheme.

Overexpression of decoy receptor 3 in hepatocellular carcinoma and its association with resistance to Fas ligand-mediated apoptosis

AIM： To characterize the expression and genomic amplification of decoy receptor 3 （DcR3） in hepatocellular carcinoma （HCC） and to evaluate the role of DcR3 in apoptosis.METHODS： We examined 48 cases of HCC for DcR3 expression by RT-PCR and DcR3 gene amplification by quantitative genomic PCR. DcR3 protein was detected by immunohistochemistry. Terminal deoxynucleotidyl transferase-mediated dUTP digoxigenin nick and labeling （TUNEL） was used to identify the apoptosis cells in tissues. Primary hepatoma cell culture and MTT test were used to evaluate the protection against FasL- and chemicalinduced apoptosis by DcR3 expression. RESULTS： DcR3 mRNA overexpression was detected in 60% HCC （29/48） patients. The occurrence of HCC was not associated with amplification of the gene. One sample base substitution was found in three sites as a sequence in Genbank. The expression of DcR3 in HCC was associated with the apoptotic index （0.067±0.04 vs 0.209±0.12, P〈0. 01）, size of mass, stage, and infiltration or metastasis （41.2% vs71.0%, 40% vs75%, 51.8% vs84.6%, P〈0. 05）. DcR3 expression could protect hepatoma cells against apoptosis induced by FasL, but not by chemicals. CONCLUSION： These data suggest that in addition to gene amplification there may be another mechanism underlying DcR3 overexpression. The effect of overexpression of DcR3 on the apoptosis of cancer cells may have direct therapeutic implications for the management of HCC.