4个矮脚鸡品系(群)6个血清蛋白(酶)位点的多态性研究

为了研究矮脚鸡的遗传多样性,采用聚丙烯酰胺凝胶电泳对兴义矮脚鸡黄羽、黑羽和麻羽及矮脚黄鸡4个品系(群)矮脚鸡6个蛋白(酶)位点进行了检测,结果表明:除Pa在矮脚黄鸡群体中表现为单态外,在其他群体中均为多态;Alb、Tf、Amy-1、ES-1、ES-2位点在各群体中均表现为多态;4个品系(群)矮脚鸡6个蛋白(酶)位点上的有效等位基因数分别为1.9769、1.8136、1.8135和1.7721,平均多态信息含量在0.3298～0.3908,呈中度多态性;4个品系(群)矮脚鸡的杂合度均较大,为0.2946～0.3680,说明它们的遗传多样性丰富,仍有较大的选育空间。

急性脑卒中后并发血管性认知障碍患者的血清半胱氨酸蛋白酶抑制剂C水平变化

目的 探讨急性脑卒中后并发血管性认知障碍（VCI）患者的血清半胱氨酸蛋白酶抑制剂（Cys）C水平的变化及其临床意义,为VCI患者的早期诊断及治疗提供依据.方法 选择2012年12月至2013年12月东营市人民医院神经内科和神经外科收治的120例急性脑卒中患者为研究对象,纳入研究组,按脑卒中后是否并发VCI将其分为VCI亚组（n=60）和非VCI亚组（n=60）;并选择同期于本院门诊体检的年龄≥50岁、无认知功能减退主诉的60例受试者纳入对照组.采用《简易精神状态评价量表》（MMSE）系统评价VCI亚组患者认知功能,并根据MMSE评分结果评价其VCI严重程度.采用全自动生化分析仪免疫比浊法检测研究组与对照组血清Cys C水平,并进行比较,同时对VCI亚组患者血清Cys C水平与VCI严重程度之间的关系进行统计学分析.本研究遵循的程序符合东营市人民医院人体试验委员会所制定的伦理学标准,得到该伦理会批准,分组征得受试对象本人的知情同意,并与之签订临床研究知情同意书.结果 VCI亚组、非VCI亚组及对照组3组受试者性别构成、年龄、受教育年限、原发性高血压、糖尿病等社会人口学资料比较,差异均无统计学意义（P＞0.05）.VCI亚组患者血清Cys C水平为（1.68±0.46） mg/L,高于非VCI亚组[（0.88±0.42） mg/L]和对照组[（0.83±0.32） mg/L],且3者比较,差异有统计学意义（F=12.99,P＜0.05）.VCI亚组患者中,VCI不同严重程度患者血清Cys C水平比较,差异有统计学意义（F=7.98,P＜0.05）.其中,MMSE评分≤13分的重度VCI患者为19例,其血清Cys C水平最高,为（1.92±0.36） mg/L;MMSE评分为≥14～20分的中度VCI患者为23例,其血清Cys C水平为（1.33±0.41） mg/L;MMSE评分为≥21～24分的轻度VCI患者为18例,其血清Cys C水平最低,为（1.09±0.28） mg/L.3者比较,差异有统计意义（F=7.98,P＜0.05）.VCI亚组患者血清Cys C水平越高,其VCI程度越严重.VCI亚组患者Cys C水平与MMSE评分的Spearmen相关系数的双变量相关分析结果示,MMSE评分与VCI患者血清CysC水平呈负相关关系（r=-0.421,P=0.039）.结论 急性脑卒中患者血清Cys C水平与其VCI的发生、发展有关.血清Cys C水平可作为急性脑卒中患者VCI早期诊断的外周血生物学标志物之一,为VCI的早期诊断与治疗提供生物学依据.

Effect of Wheat Based Diet Supplemented with Xylanase on Blood Sugar and Total Protein in Serum of Geese

[Objective] The effects of wheat based diet supplemented with xylanase on blood sugar and total protein in serum of geese were studied. [ Method ] By using the randomized design of single factor, the 1-day-old healthy goslings were divided into 6 groups and fed with corn based diet, wheat based diet and wheat based diet supplemented with xylanase at different concentrations respectively, the contents of blood sugar and total protein in serum were determined. [ Result] The wheat based diet supplemented with xylanase could increase the blood sugar and total protein in serum of geese and wheat based diet supplemented with 0.2% xylanase generated the best effect, which was higher than those of corn based diet group. As for the concentration of protein in serum, wheat based diet supplemented with 0.2% xylanase was significantly different from corn based diet and wheat based diet. [ Conclusion] The wheat based diet supplemented with xylanase could enhance geese production.

Evaluation of serum catnepsin B and D in relation to diniGopatnological staging of colorectal cancer

AIM: Proteolytic degradation of the extracellular matrix facilitates cancer invasion and promotes metastasis. The study aims at evaluation of preoperative and postoperative serum cathepsins B and D levels in correlation with selected anatomoclinical features of colorectal cancer.METHODS: Blood samples were collected from 63colorectal cancer patients before curative operation of the tumor 10 d later. Blood that was obtained from 20healthy volunteers, served as a control. The activity of cathepsin B was measured with Bz-DL-arginine-pNA as a substrate at pH 6.0, while cathepsin D activity was determined with urea-denatured hemoglobin (pH 4.0).RESULTS: The preoperative and postoperative activities of cathepsin B were significantly (P＜0.00001) lower in serum of colorectal cancer patients than in control group.However, postoperative values of this protease were significantly increased in comparison with preoperative ones (P = 0.031). Activity of cathepsin D appeared to be significantly higher in colorectal cancer sera (P＜0.00001)compared with controls. No statistically significant differences between preoperative and postoperative activity of cathepsin D were noted (P = 0.09). We revealed a strong linkage of cathepsins' levels with lymph node status and pT stage of colorectal cancer.CONCLUSION: Blood serum activities of cathepsin B and D depend on the time of sampling, tumor size and lymph node involvement. Significantly, increased activity of cathepsin D could indicate a malignant condition of the large intestine. In our work, the serum postoperative decrease of cathepsin B activity appears as an obvious concomitant of local lymph node metastasis-the wellknown clinicopathological feature of poor prognosis.

Effect of Pore-size of Mesoporous SBA-15 on Adsorption of Bovine Serum Albumin and Lysozyme Protein

In this paper,the effects of pore-size of SBA-15 on the adsorption kinetics and equilibrium of large protein molecules Bovine serum albumin(BSA)and lysozyme(LYS)have been investigated.The mesoporous molecular sieve SBA-15 with six different pore sizes were synthesized with P123 triblock copolymer as the template agent,and 1,3,5-trimethylbenzene(TMB)and isopropyl alcohol as the pore-expanding agent.The samples were characterized by N2 adsorption/desorption,Scanning Electron Microscopy(SEM),Transmission Electron Microscopy(TEM)and X-Ray Diffraction(XRD).It is found that BSA and LYS were adsorbed rapidly on SBA-15 materials with large pores.The BSA adsorption capacity of sieve with the pore diameter of 21.4 nm reached 500 mg·g-1 within 25 minutes.However,if the pore diameter was smaller than 14 nm,the BSA adsorption capacity of the sieve was only about 220 mg·g-1.The adsorption equilibrium data fits in the Langmuir model,where the coefficient of effective use of specific area of mesoporous molecular sieve was found to be 0.03,0.18,0.37 and 0.48,corresponding to the pore diameter of 10.1 nm,13.2 nm,15.4 nm and 21.4 nm,respectively.The equilibrium loading amount of LYS on SBA-15 materials with pore size of 15.4 nm could be up to 1000 mg·g-1.The coefficient of effective use of surface area of mesoporous molecular sieve with diameter of 3.9 nm,7.4 nm,10.1 nm,13.2 nm and 15.4 nm was 0.10,0.47,0.56,0.71 and 0.79,respectively.It is also noted that greater pore size of mesoporous molecular sieve would lead to a higher coefficient of effective use of surface area.

Acute pancreatitis in aging animals:Loss of pancreatitis-associated protein protection?

AIM:To investigate the effect of age on severity of acute pancreatitis(AP) using biochemical markers,histology and expression of the protective pancreatitisassociated proteins(PAPs).METHODS:AP was induced via intraductal injection of 4% sodium taurocholate in young and old rats.Sera and pancreata were assayed at 24 h for the parameters listed above;we also employed a novel molecular technique to assess bacterial infiltration using polymerase chain reaction to measure bacterial genomic ribosomal RNA.RESULTS:At 24 h after induction of AP,the pancreata of older animals had less edema(mean ± SE histologic score of young vs old:3.11 ± 0.16 vs 2.50 ±-0.11,P < 0.05),decreased local inflammatory response(histologic score of stromal infiltrate:3.11 ± 0.27 vs 2.00 ± 0.17,P < 0.05) and increased bacterial infiltration(174% ± 52% increase from sham vs 377% ± 4%,P < 0.05).A decreased expression of PAP1 and PAP2 was demonstrated by Western blotting analysis and immunohistochemical staining.There were no differences in serum amylase and lipase activity,or tissue myeloperoxidase or monocyte chemotactic protein-1 levels.However,in the most-aged group,serum C-reactive protein levels were higher(young vs old:0.249 ± 0.04 mg/dL vs 2.45 ± 0.68 mg/dL,P < 0.05).CONCLUSION:In older animals,there is depressed PAP expression related to a blunted inflammatory response in AP which is associated with worsened bacterial infiltration and higher C-reactive protein level;this may explain the more aggressive clinical course.

Protein Templated Au-CuO Bimetallic Nanoclusters toward Neutral Glucose Sensing

In this study,the application of bovine serum albumin(BSA)as a carrier to glucose-sensitive materials for the detection of glucose was proposed.Au-Cu O bimetallic nanoclusters(Au-Cu O/BSA)were prepared using BSA as a template,the new sensing material(Au-Cu O/BSA/MWCNTs)was synthesized by mixing with multi-walled carbon nanotubes(MWCNT)and applied to non-enzymatic electrochemical sensors to detect glucose stably and effectively under neutral condition.The scanning electron microscopy was used to investigate the morphology of the synthesized nanocomposite.The electrochemical properties of the sensor were studied by cyclic voltammetry.Glucose detection experiments show that Au-Cu O/BSA/MWCNTs/Au electrode has good glucose detection ability,stability,accuracy,repeatability,and high selectivity in neutral environment.Unlike existing glucose-sensitive materials,due to the use of BSA,the composite material is firmly fixed to the electrode surface without a Nafion solution,which reduces the current blocking effect on the modified electrode.The composite materials can be effectively preserved for extremely long periods,higher than 80%activity is maintained at room temperature in a closed environment for 3 to 4 months,due to the special effects of BSA.In addition,the feasibility of using BSA in glucose-sensitive materials is confirmed.

Serum manganese superoxide dismutase and thioredoxin are potential prognostic markers for hepatitis C virus-related hepatocellular carcinoma

AIM:To evaluate the clinical significance of oxidative stress markers in patients with hepatitis C virus(HCV)related hepatocellular carcinoma(HCC).METHODS:Sixty-four consecutive patients who were admitted to Kagoshima University Medical and Dental Hospital were enrolled in this retrospective study.All patients had chronic liver disease(CLD) due to infection with HCV.Thirty patients with HCV-related HCC,34 with HCV-related CLD without HCC(non-HCC),and 20 healthy volunteers(HVs) were enrolled.Possible associations between serum manganese superoxide dismutase(MnSOD) and thioredoxin(TRX) levels and clinical parameters or patient prognosis were analyzed over a mean follow-up period of 31.7 mo.RESULTS:The serum MnSOD levels were significantly higher in patients with HCV-related HCC than in patients without HCC(P = 0.03) or HVs(P < 0.001).Similarly,serum TRX levels were also significantly higher in patients with HCV-related HCC than in patients without HCC(P = 0.04) or HVs(P < 0.01).However,serum levels of MnSOD and TRX were not correlated in patients with HCC.Among patients with HCC,the overall survival rate(OSR) was lower in patients with MnSOD levels ≥ 110 ng/mL than in patients with levels < 110 ng/mL(P = 0.01),and the OSR tended to be lower in patients with TRX levels < 80 ng/mL(P = 0.05).In addition,patient prognosis with HCC was poorest with serum MnSOD levels ≥ 110 ng/mL and serum TRX levels < 80 ng/mL.Furthermore,a multivariate analysis using a Cox proportional hazard model and serum levels of five factors(MnSOD,prothrombin time,serum albumin,serum α-fetoprotein(AFP),and serum des-γ-carboxy prothrombin) revealed that MnSOD levels ≥ 110 ng/mL(risk ratio:4.12,95% confidential interval:1.22-13.88,P = 0.02) and AFP levels ≥ 40 ng/mL(risk ratio:6.75;95% confidential interval:1.70-26.85,P < 0.01) were independent risk factors associated with a poor patient prognosis.CONCLUSION:Serum MnSOD and TRX levels are potential clinical biomarkers that predict patient prognosis in HCV-related HCC.

Serum metrix metalloproteinase-9 combined with homocysteine, IL-6, TNF-α, CRP, HbA11c and lipid profile in the incipient diabetic nephropathy with or without macrovascular diseases

Objective ： To evaluate the changes of serum matrix metalloproteinase-9 （romp-9） in patients of incipient diabetic nephropathy with or without macrovascular disease and to analyze the factors associated with homocysteine（hcy）, interleukin-6（IL-6）, tumor necrosis factor-alpha （TNF-α）, highly sensitive C-reactive protein （hsCRP）, HbAlc and lipid profile in those patients in order to know whether this marker or other factors are more important to induce diabetic macrovascular disease. Methods： Type 2 diabetes mellitus（T2DM） subjects with incipient diabetic nephropathy with or without macrovascular disease were selected for participation and divided into 2 groups. The patients in group 1 （n= 38） used insulin, and patients in group 2 （n=34） were treated with an oral antidiabetic drug. Then serum mmp-9, hey, IL-6 and TNF-α in these patients were measured, and compared to the healthy subjects as control （n= 16）. The results were analyzed by SPSS13. Results： Serum romp-9 and hcy of the patients having incipient diabetic nephropathy with macrovascular disease were higher than that of patients without macrovascular disease （P〈0.01）. For insulin-injected patients, whether they accompanied with macrovascular diseases or not, the serum levels of romp-9, hcy, IL-6 and TNF-α were all lower, but no significant statistics compared with non-insulin used patients or the healthy subjects. The serum level of romp-9 was more correlated with the serum hcy in antidiabetic drug used patients. （P〈0. 000） Conclusion： The serum level of romp-9 plays an important role of pathogenesis in the macrovascular disease in the incipient diabetic patients, and the serum level of hcy also can reflect the severely degree of macrovascular disease in these patients, insulin can reduce these markers.

Prognostic values of serum LDH and β2-MG in patients with non-Hodgkin’s lymphoma

Objective： To evaluate the clinical significance of quantitative determination of serum lactic dehydrogenase （LDH） and beta 2-microgtobulin （β2-MG） in patients with non-Hodgkin＇s lymphoma （NHL）. Methods： Date of the 65 patients from 2003 to 2004 were retrospectively analyzed, relationship between LDH and β2-MG activities and tumor phase, effect of treat- ment as well as prognosis were studied. Results： The levels of LDH and β2-MG in the advanced stage, B group and bone marrow involvement were obviously higher than their comparison counterpart and the difference was significant （P 〈 0.05）. There was no significant difference between the high LDH group and normal LDH group in the two-year survival rate. But the difference in the four-year survival rate was significance. Conclusion： The level of LDH and β2-MG are valuable for clinical stage, prognosis and evaluation of therapeutic response in patients with NHL.

A novel organic-inorganic hybrid monolith for trypsin immobilization

In proteomics, attention has focused on various immobilized enzyme reactors (IMERs) for the realization of high throughput digestion. In this report, a novel organic-inorganic hybrid monolith based IMER was prepared in a 100 μm i.d. capillary with 3-glycidoxypropyltrimethoxysilane (GLYMO) as the monomer and tetraethoxysilane (TEOS) as the crosslinker. Trypsin immobilization was achieved via the reaction between vicinal diol groups, which were obtained from hydrolysis of epoxy groups, and the amino groups of trypsin. Bovine serum albumin was digested thoroughly by this IMER in 47 s. After micro-reverse phase liquid chromatography-tandem mass spectrometry (μRPLC-MS/MS) analysis and database searching, beyond 35% sequence coverage was obtained, and the result was comparable to that of 12 h in solution digestion. The present IMER has potential for high throughput digestion.

Dynamic changes of serum cholinesterase activity after severe trauma

Objective： The aim of the present study was to examine dynamic changes in serum cholinesterase （ChE） activity during early-stage severe trauma and the clinical significance of these changes. Methods： This prospective, observational study included 81 patients with severe trauma who were treated between October 2011 and April 2013 in the emergency intensive care unit （EICU） of a university-affiliated, tertiary-care, grade A general hospital in China. Serum ChE activity was measured on Days 1, 3, and 7 post-injury. The correlation of dynamic changes in serum ChE activity with trauma severity and prognosis was assessed. Correlations between changes in serum ChE activity after injury and albumin （ALB）, prealbumin （PAB）, transferrin （TRF）, and C-reactive protein （CRP） levels were also analyzed Results： Serum ChE activity in trauma patients was 42.3%-50.2% lower on Days 1, 3, and 7 compared with the control （P〈0.001 for all time points）, and it continued to decrease after Day 7 in both the survival and death subgroups. In the subgroup with an injury severity score （ISS） of 〈25, serum ChE activity initially decreased, but eventually increased. However, activity decreased continuously in the ISS〉25 subgroup. ChE activity was significantly lower in both the death and the ISS〉25 subgroups than in the survival and ISS〈25 subgroups on Days 1, 3, and 7 after injury. Activity was negatively correlated with ISS and acute physiology and chronic health evaluation Ill （APACHE III） at all time points. When comparing the receiver operating characteristic （ROC） curves for predicting prognosis, the area under the curve （AUC） in the dot of serum ChE was similar to the AUCs in plots of ISS and APACHE Ⅲ, but significantly smaller than the AUC in the plot of the trauma and injury severity score （TRISS）. Serum ChE activity was positively correlated with ALB, PAB, and TRF at all time points post-injury. Activity was not significantly correlated with CRP on Day 1, but was significantly and negatively correlated with CRP on Days 3 and 7. Conclusions： There is a significant decrease in serum ChE activity after severe trauma. Serum ChE may be regarded as a negative acute phase protein （APP） and the dynamic changes in serum ChE may be useful as an auxiliary indicator for evaluating trauma severity and predicting prognosis.

Changes of balance between proteinase and their inhibitors in blood of pigs with high-velocity missile wounds

Objective: To study the effect of imbalance between lysosomal enzymes and their inhibitors in blood on disturbance of the local and whole body after trauma.Methods: The dynamic changes of lysosomal enzymes and proteinase inhibitors were studied in 12 pigs with femoral comminuted fractures in both hind limbs caused by high velocity missiles. Four normal pigs served as controls.Results: After injury, the activity of Cathepsin D in arterial plasma increased gradually and reached the highest level at 8 hours, acid phosphatase in serum began to increase at 12 hours and the value of serum elastase did not change significantly. The level of αl-antitrypsin, a proteinase inhibitor in plasma, decreased significantly in the early stage after injury [ 73.5% ±6.4% and 81.0% ±5.1% of the baseline value (1.67 μmol · ml-1 · min-1 ± 0.29 μmol · ml-1 · min-1) at 1 and 2 hours after injury, respectively, P < 0.05 ], then increased gradually and was higher than the baseline value at 12 hours after injury.Conclusions: Imbalance between lysosomal enzymes and proteinase inhibitors occurs soon after injury, which might result in continuous tissue damage and play an important role in the disturbance of general reaction after injury.