一例母及其亲子DNA指纹与血清蛋白、酯酶遗传的研究

采用ＤＮＡ指纹分析和聚丙烯酰胺凝胶电泳法，对一例与公驴交配生育了后代的母后代进行了亲缘鉴定和血清蛋白、酯酶遗传的分析。结果可以认定其亲子关系并证实母骡生育的事实。虽然本例母的后代在Ｐｒ、Ａｌ、Ｐａ和Ｈｂ、Ｅｓ等基因座位上的基因表达倾向于驴，但其外貌仍明显地带有种间杂种的特征。因此，尚不能简单地认为其已“回归”为纯种的驴。

Genes Expression Profile Difference in Peripheral Blood Between Esophageal Carcinoma Patients and Normal Subjects

Objective： To study the genes expression profile differences in the peripheral blood between esophageal carcinoma patients and normal subjects using the gene chip technique and screen out the esophageal early conceration associated genes. Methods： The total RNA was extracted and purified in the peripheral blood obtained from the patients with esophageal carcinoma and normal subjects. The first strand of cDNA was synthesized through retro-transcription and labeled with Cy5 and Cy3 fluorescence as probes. The mixed probes were hybridized with a piece of 4096 double dot human whole gene chip. The acquired image was analyzed by microarrav suite software using a digital computer, and the intensity of ttuorescence signal and its ratio were calculated. Results： A total of 92 genes were screened out and its expression difference was more than 2 times in the peripheral blood between the patients with esophageal carcinoma and normal subjects. Among these, the expression difference of 36 genes was more than 3 times. Two human urokinase plasminogen activator surface receptor （UPAR） genes, 80K-L protein gene, human protein tyrosine-phosphatase gent arid proto-oncogene protein mRNA were significantly up-regulated, while the collagen V type （α-2 gene was markedly down-regulated. Conclusion： 80K-L protein gene, tyrosinephophatase gene, proto-oncogene protein arid the collagen V type α-2 gene might be associated with the ontogenesis, development and its metastasis in the esophageal carcinoma. The UPAR gene may play important roles in the diagnosing the micrometastasis in the peripheral blood of esophageal carcinoma.

Synthesis and Adsorption Study of BSA Surface Imprinted Polymer on CdS Quantum Dots

A new bovine serum albumin （BSA） surface imprinting method was developed by the incorporation of quantum dots （QDs） into molecularly imprinted polymers （MIP）, which can offer shape selectivity. Preparation and adsorption conditions were optimized. Physical appearance of the QDs and QDs-MIP particles was illustrated by scanning electron microscope images. Photoluminescence emission of CdS was quenched when rebinding of the template.The quenching of photoluminescence emissions is presumably due to the fluorescence resonance energy transfer between quantum dots and BSA template molecules. The adsorption is compiled with Langmuir isotherm, and chemical adsorption is the rate-controlling step.The maximum adsorption capacity could reach 226.0 mg/g, which is 142.4 mg/g larger than that of undoped BSA MIP. This study demonstrates the validity of QDs coupled with MIP technology for analyzing BSA.

Expression and purification of the complete PreS region of hepatitis B Virus

AIM: To express the complete PreS region of HBV in E.coli with good solubility and stability, and to establish an effective method for purification of the recombinant PreS protein. METHODS: The complete PreS region (PreS1 and PreS2) was fused into a series of tags including glutathione S-transferase (GST), dihydrofolate reductase (DHFR), maltose binding protein (MBP), 6× histidine, chitin binding domain (CBD), and thioredoxin, respectively. Expression of recombinant PreS fusion proteins was examined by SDS-PAGE analysis and confirmed by Western blot. Two fusion proteins, thio-PreS, and PreS-CBD, with desirable solubility and stability, were subjected to affinity purification and further characterization. RESULTS: Recombinant PreS fusion proteins could be synthesized with good yields in E.coli. However, most of these proteins except for thio-PreS and PreS-CBD were vulnerable to degradation or insoluble as revealed by SDS-PAGE and Western blot. Thio-PreS could be purified by affinity chromatography with nickel-chelating sepharose as the matrix. However, some impurities were also co-purified. A simple freeze-thaw treatment yielded most of the thio-PreS proteins in solution while the impurities were in the precipitate. Purified thio-PreS protein was capable of inhibiting the binding of HBV virion to a specific monoclonal antibody against an epitope within the PreS1 domain. CONCLUSION: Increased solubility and stability of the complete PreS region synthesized in E.coli can be achieved by fusion with the thioredoxin or the CBD tag. A simple yet highly effective method has been established for the purification of the thio-PreS protein. Purified thio-PreS protein likely assumes a native conformation, which makes it an ideal candidate for studying the structure of the PreS region as well as for screening antivirals.

Characterization of lipid parameters in diabetic and non-diabetic atherosclerotic patients

Background ＆ Objective The relationship between lipid profile perturbation and diabetes associated complications has long been an area of interest. Dyslipidemia is a potent predictor of cardiovascular morbidity and mortality in diabetic patients. The aim of present study was to investigate relationship between aging and lipid profiles in diabetic and non-diabetic atherosclerotie patients. Methods Five hundred and seventy six individuals （45-75 year age） participated in this study. Among these, 192 were having history of diabetes mellitus and atherosclerosis. Individuals are categorized on the base of health （normal, non-diabetic atheroselerosis, diabetic atherosclerosis） and age （45-55 years, 56~55 years, and 66-75 years）. All the participants were subjected to the procedures like a detailed history, biochemical analysis for fasting blood sugar, hemoglobin Alc, total cholesterol （TC）, triglycerides （TG）, low-density lipoprotein-（LDL）, very low-density lipoprotein （VLDL） and high-density lipoprotein （HDL）. All these parameters were compared between diabetic and non-diabetic atherosclerotic patients of all three age groups. TC/HDL and LDL/HDL were also calculated. Results Diabetic atherosclerotic individuals （both males and females） had high level of TC, TG, LDL, VLDL and low level of HDL in comparison to non-diabetic atherosclerotie and normal control individuals. Among all three age groups, lipoprotein abnormality was observed to be more frequent in females than males. There was a significant increase in TC/HDL and LDL/HDL ratio in diabetic atherosclerotic subjects compared to age and sex matched non-diabetic atherosclerotic and normal control. Conclusions Degree of dyslipidemia increases with increase in age in both genders. Female are more prone to diabetic dyslipidemia and hence have more risk of developing atherosclerosis with increasing age.

Expressions of MVD, VEGF, Ki67 in Residual Prostate Cancer after Cryoablation

OBJECTIVE To analyze the effects of cryoablation on the mice bearing Rm-I prostate cancer through detecting tumor angiogenesis and cancer cell proliferation in the mice after cryoablation, and to explore the effects of cryoablation on vascular endothelium growth factor （VEGF）, Ki67 protein expression and microvessel density （MVD） in the mice bearing prostate cancer. METHODS Sixty Rm-1 mouse models of prostate cancer were established. Experimental mice were randomized into 2 groups： the cryoablation group （n = 30） and the control group （n = 30）. After file therap）4 tumor tissues of the mice in group A and B were obtained at day 0 （without cryoablation）, 1st, 3rd, 5th, 7th, 14th day, respectivelj6 after cryoablation, and the expressions of MVD, VEGF and Ki67 proteins were detected at the same time points. RESULTS The expressions of MVD, VEGF and Ki67 proteins in group A were decreased. The lowest values of the factors were detected on the 3rd day after cryoablation, and increased slowly after that. The expressions of MVD, VEGF and Ki67 proteins in the control group were not changed. Significant changes of the expressions of MVD, VEGF and Ki67 proteins in the group A were found at different time points. Correlation analysis suggested a positive correlation between the expressions of VEGF and MVD proteins （r = 0.8793）, a positive correlation between the expressions of Ki67 and MVD proteins （r = 0.7614）, and a positive correlation between the expressions of VEGF and ki67 proteins （r = 0.6921）. CONCLUSION After argon-helium cryoablation treatment for the mice bearing prostate cancer, the expressions of MVD, VEGF and Ki67 proteins in local tumor were reduced on the 1st day. The lowest values of the factors were detected on the 3rd day after cryoablation, and then increased after that. Cryoablation combined with other modalities of treatment may effectively improve the treatment effects of cryoablation for prostate cancer.

NF-E2:a Novel Regulator of Alpha-hemoglobin Stabilizing Protein Gene Expression

Objective To investigate whether α-hemoglobin stabilizing protein (AHSP), the α-globin-specific molecular chaperone, is regulated by erythroid transcription factor NF-E2. Methods We established the stable cell line with NF-E2p45 (the larger subunit of NF-E2) short hairpin RNA to silence its expression. Western blot, real-time polymerase chain reaction, and chromatin immunoprecipitation (ChIP) analysis were performed to detect the expression of AHSP, the histone modifications at AHSP gene locus, and the binding of GATA-1 at the AHSP promoter with NF-E2p45 deficiency. ChIP was also carried out in dimethyl sulfoxide (DMSO)-induced DS19 cells and estrogen-induced G1E-ER4 cells to examine NF-E2 binding to the AHSP gene locus and its changes during cell erythroid differentiation. Finally, luciferase assay was applied in HeLa cells transfected with AHSP promoter fragments to examine AHSP promoter activity in the presence of exogenous NF-E2p45. Results We found that AHSP expression was highly dependent on NF-E2p45. NF-E2 bound to the regions across AHSP gene locus in vivo, and the transcription of AHSP was transactivated by exogenous NF-E2p45. In addition, we observed the decrease of H3K4 trimethylation and GATA-1 occupancy at the AHSP gene locus in NF-E2p45-deficient cells. Restoration of GATA-1 in G1E-ER4 cells in turn led to increased DNA binding of NF-E2p45. Conclusion NF-E2 may play an important role in AHSP gene regulation, providing new insights into the molecular mechanisms underlying the erythroid-specific expression of AHSP as well as new possibilities for β-thalassemia treatment.

Expression differences of serum prealbumin in benign and malignant colorectal tumors

Objective: The aim of the study was to investigate the expression differences of serum prealbumin in patients with benign and malignant colorectal tumors and its clinical significance. Methods: The concentrations of total protein, albumin, prealbumin, hemoglobin of 113 colorectal cancer patients(cancer group) and 87 colorectal adenomas(adenoma group) were tested in Yixing Hospital Affiliated to Jiangsu University(China) during August 2013 to December 2013. Then the differences between the two groups were compared. Results: In colorectal cancer patients, the concentrations of serum prealbumin in 39/113 cases, total protein in 16/113 cases, albumin in 38/113, hemoglobin in 32/113 were lower than normal ranges. While, in colorectal adenoma patients, the concentrations of serum prealbumin in 4/87 cases, total protein in 2/87, albumin in 1/87, hemoglobin in 2/87 were below the detection limit. Comparative analysis showed that, average expression levels of serum prealbumin, albumin, total protein, hemoglobin in colorectal cancer patients were lower than those of colorectal adenoma patients, the difference was statistically significant(P < 0.05), and colorectal cancer patients were more likely to have lower levels of above indicators(P < 0.05). Conclusion: Compared to colorectal adenoma patients, patients with colorectal cancer have lower average expression levels, and were easier to have lower expression levels of serum albumin, albumin, total protein and hemoglobin, which suggest that colorectal cancer patients are more likely to have metabolic change, and clinic notable.

Expression of Thrombospondin-1 is Correlated with Microvessel Density in Prostate Cancer

OBJECTIVE To observe the expression of thrombospondin-1 ( TSP-1) in prostate cancer, and examine its expression in relation to angiogenesis. METHODS The expression of TSP-1 and microvessel density (MVD) were studied in 22 prostate cancer patients by using immunohistochemistry. RESULTS Positive expression of the TSP-1 protein was detected in 16 (72.7%)of the 22 cases. Most of the positive staining for TSP-1 was seen in the cytoplasm of the cancer cells, but some was in the extracellular matrix. The mean MVD in the 22 prostate cancer cases was 71.21±31.14 vessels per 100 high field of vision. Tumors with an elevated expression of TSP-1 showed a high MVD resulting in a correlation between TSP-1 immunopositivity and microvessel density that was highly significant (r= 0.54, P=0.009). CONCLUSION TSP-1 is strongly expressed in most prostate cancers and is associated with neovascularization. Therefore TSP-1 is a likely contributor to the extensive neovascularization in prostate cancer and increased TSP-1 expression might participate in an angiogenic phenotype.

Radiofrequency induction on sodium/iodide symporter expression of thyroid cancer

Objective; The aim of this study was to investigate the effects of radiofrequency treatment on sodium/iodide symporter expression of thyroid cancer ceils. Methods： In 29 thyroid cancer patients with low or no expression of soda ／ iodide symporter, the radio frequency combined 1311 therapy was used, the whole-body scintigraphy and serum Ig were detected before and after the radiofrequency treatment. Results： The whole-body scintigraphy showed that 4 cases （4/29） before radiofrequenc_y treatment had positive iodine uptake, 19 cases （19/29） two weeks after radiofrequency treatment had the positive iodine uptake, 12 cases （12/29） four weeks after radiofrequency treatment had the positive iodine uptake. Four weeks after radiofrequency treatment, 5 cases had increased serum Ig levels, 17 cases had decreased serum Ig levels, 7 cases showed no change. 25 cases （25/29） were effective, 15 cases （15/29） were cured. Conclusion： The radiofrequency induced the non-expressed the sodium/iodide symporter of thyroid cancer cells regain the iodine intake ability, it improved the clinical efficacy of 131I therapy in dedifferentiated thyroid cancer.

Effects of aerobic training on serum paraoxonase activity and its relationship with PON1-192 phenotypes in women

Background：Paraoxonase 1（PON1） is an antioxidant enzyme that protects high-density lipoprotein（HDL） and low-density lipoprotein against oxidation.Limited studies have addressed the influenc of exercise on PON1 activity and its relationship with PON1 phenotypes.We investigated relationships between PON1-192 phenotypes,PON1 activity,aerobic exercise,and blood lipid and lipoprotein concentrations in middle-aged women.Methods：An exercise group（n=50） engaging in regular aerobic exercise and a control group（n=41） were selected from a subset of 300 Caucasian women that met the inclusion criteria.Serum PON1,salt-stimulated PON1（SSPON1）,and arylesterase（ARE） activities;cholesterol levels and ARE activities of total HDL and HDL subgroups（HDLs）（supernatants obtained by polyethylene glycol）;and blood lipid and lipoprotein concentrations were determined by standardized enzymatic methods.PON1-192 QQ（low activity）,QR（moderate activity）,and RR（high activity） phenotype groups were define using serum SSPON1/ARE activity ratios.The R-carries（RC） phenotype group consisted of the QR and RR groups combined.Results：All lipid and lipoprotein concentrations were greater in the exercise group than in the control group.Regardless of phenotype,no significan differences were observed between the exercise and control groups in terms of serum PON1,SSPON1,or ARE activity associated with HDLs（p〉 0.05）,whereas PON1 activities in QQ-phenotyped women in the exercise group were significant y higher than those in the control group（p〈0.01）,but not the RC group.A statistically significan interaction between PON1 phenotypes（QQ and RC groups） and exercise（exercise and control groups） on PON1 activity was found.Conclusion：These results showed that a regular aerobic exercise program can improve PON1 activity depending on PON1-192 phenotype,but not on lipid and lipoprotein levels,in middle-aged Turkish women.

ahospholipase Applications in Cheese Production

Phospholipase is an enzyme that hydrolyzes phospholipids releasing a variety of products, like for example lyso-phospholipids, free fatty acids, di-acylglycerols, choline phosphate and phosphatidates, depending on the site of hydrolysis. In cheese production, lysophospholipids act as surface-active agents in the cheese curd, helping emulsification of water and fat during processing and reducing syneresis. Phospholipases are more specific and have little or no activity toward di- or triglycerides. As a result of phospholipid hydrolysis, flavor defects do not occur due to the main formation of palmitic, oleic, and stearic acids, which are non-volatile short chains fatty acids. According to the scientific studies the use of phospholipase is able to increase the yield of cheese and reduce the environmental impacts of cheese production. Protein and fat largely determine cheese yield. Depending on the milk composition, 75% to 78% of milk protein and 85% to 95% of milk fat are entrapped in the cheese curd. The remaining protein and fat are lost in the whey and, to a lesser extent, in the brine. Crucially in the production of pasta filata cheese fat losses occur in the hot stretching step, where the fresh curd is molded and stretched in hot water. The lysophospholipid-casein complexes should be studied to understand the mechanism leading to cheese yield improvements.

Effects of dendritic cell vaccines on hematogenous micrometastasis of bladder cancer carrying for PBL-SCID mice

Objective： To study the effect of dendritic cells loaded with whole tumor antigen on hematogenous micrometastasis of bladder cancer model in hu-PBL-SCID mice. Methods： T24-3 ceil subset was selected from human bladder transitional cell carcinoma T24 cell line by Boyden chamber system. The SCID mice intraperitoneally injected with 4 × 10^7 hu-PBL and subcutaneously injected with 3 × 10^6 T24-3 cells were named hu-PBL-T24-3-SCID model. Human IgG level in the blood plasma of mice was detected by ELISA, and human CD3^＋, CD4^＋, CD8^＋ T cells in blood and spleen cells of mice were detected by FCM analysis for human immune reconstruction study. Human CK20 mRNA expression in mice peripheral blood was detected by RT-PCR to investigate metastasis of tumor cells. The PBMCs were isolated from human peripheral blood, and were induced into DCs by co-culture with rhGM-CSF and rhlL-4 in vitro. The DC vaccines were produced by co-culturing with whole tumor antigen which was purified through freezing and melting T24-3 cell subset. After T24-3 cells injected into SCID mice for 5 weeks, the mice were treated with DC vaccines. Results： All mice were initially treated at 5th week. The expression of CK20 mRNA in peripheral blood of DC vaccines treated mice was the lowest. There was 2 mice showing CK20 mRNA expression and 3 mice with metastasis tumor in PBS group. MMP-7 mRNA expression in tumor tissues of DC vaccines treated mice was statistically lower than that of PBS group （P 〈 0.01）. Conclusion： DC vaccines have a good effect on hu-PBL-SCID mice bladder cancer model by reducing hematogenous micrometastasis.

HIF-1 inhibitors as anti-cancer therapy

Hypoxia is a hallmark of solid tumors.Hypoxia increases the progression of malignancy and metastasis by promoting angiogenesis and triggering the over-expression of various protein products critical for tumor growth.The transcription factor HIF-1 mediates cellular response to hypoxia by promoting processes,such as glycolysis and angiogenesis.Clinical evidence has demonstrated that expression of HIF-1 is strongly associated with poor patient prognosis and activation of HIF-1 contributes to malignant behavior and therapeutic resistance.Therefore,HIF-1 is a viable target for cancer therapy.This review summarizes agents that have been described in the literature as HIF-1 inhibitors.The majority of these compounds are indirect inhibitors of HIF-1.

Expression of hunchback during oogenesis and embryogenesis in Locusta migratoria manilensis(Meyen)

hb(hunchback) is a contributing factor in anteroposterior axial patterning of insects.Although the hb function in Locusta migratoria manilensis has been investigated,its expression pattern remains unknown.Here,the mouse polyclonal antibody was produced against Hb fusion protein,and then its expression pattern during oogenesis and embryogenesis of L.migratoria manilensis was examined by immunohistochemical staining.Hb protein was detected in the oocyte nucleus which was positioned centrally within the developing oocyte.The oocyte nucleus gradually moved to the posterior end of the egg along with the oocyte maturing.In freshly laid eggs,Hb formed gradient at the posterior end of the egg,and then hb was expressed as a band in the middle of the blastodisc.As the blastodisc differentiated into the head and trunk,the expression region became wide,which would develop into spatial gnathal and thoracic segments.With abdominal segmentation,the expression domain in the gnathal and thoracic region became faint and eventually faded out,while the Hb expression domain appeared at the posterior growth zone in a discontinuous expression manner.The hb expression pattern of L.migratoria manilensis is greatly similar to that of other locusts,such as Schistocerca americana and another L.migratoria.Compared with other insects,hb expression is conserved in the gnathal and thoracic domains,while divergent in oogenesis and abdomen.

Effect of aqueous extract of Sanweitanxiang powder on calcium homeostasis protein expression in ischemic-reperfusion injury rat heart

OBJECTIVE: To investigate the underlying mechanism of reduced myocardial ischemia-reperfusion (I/R) injury in rats using the traditionalTibetan medicine Sanweitanxiang powder (SWTX). METHODS: Rats were randomly divided into six groups (n=10) as follows: (a) propranolol dinitrate control group, given propranolol dinitrate 0.02 g/kg for 10 days before I/R, (b) SWTX with a high dose group, given SWTX 1.5 g/kg for 10 days before I/R, (c) SWTX with a medium dose group, given SWTX 1.25 g/kg for 10 days before I/R, (d) sham group (Sham), in which the rat heart was exposed by pericardiotomy but without I/R, (e) SWTX with a low dose group, given SWTX 1.0 g/kg for 10 days before I/R, and (f) I/R injury group. Rats were intragastrically pretreated with propranolol dinitrate orSWTX. After that, the operation to cause ischemia and reperfusion was conducted.The histopathologic changes of rat hearts were observed by hematoxylin and eosin staining and transmission electron microscopy. Ca2+ homeostasis protein expression was determined by western blot. RESULTS: After SWTX pretreatment, the development of ultrastructural pathological changes from IR injury was attenuated. A decrease in the expression of B-cell lymphoma 2 associated X protein, and an increase in the expression of B-cell lymphoma 2 were observed. An increased activation of extracellular signal regulated kinases were found. Compared with the sham group, the expression of sarcoplasmic reticulum calcium-ATPase, phospholamban, and calsequestrin were all up-regulated after pretreatment with SWTX. CONCLUSION: The protective mechanism of SWTX pretreatment on myocardial I/R injury might be related to its effect on maintaining the balance of calcium homeostasis in rat heart.