Enzyme immobilization is the core technology of biocatalysis. Over the past few decades, enzyme immobilization research mainly focused on single enzyme immobilization. In recent years, multi-enzyme immobilization attracts more and more attention as it could increase the local concentration of reaction and improve the reaction yield. In this review, a summary of the recent progress, together with our research, is presented. Special emphasis is placed on four methods in multi-enzymes co-immobilization, namely, the nonspecific covalent co-immobilization, the nonspecific non-covalent co-immobilization, the non-covalent encapsulation co-immobilized and the site specificity co-immobilized. Finally, some industrial uses of immobilized multi-enzymes were addressed and the application prospect of multi-enzyme immobilization was highlighted.
Biocatalysis ; Enzymes, Immobilized

Objective:To probe into the expression and the clinic significance of LEA on colorectal carcinoma. Methods:The expression of LEA and CEA in 140 colorectal cancer specimens and 100 colorectal non-cancerous specimens had been detectd by immunohistcchemistry S-P method.Results:The expression of LEA was relative to tumor differentiation degree and exhibits higher selectivity in high differentiation ade-nocarcinoma ( P 0.05) . The positive rate of LEA in adenoma was much higher than surrounding non-cancerous mucosa and normal mucosa. In normal mucosa the positive rate of LEA was obviously lower than that of CEA ( P

Objective　To observe the ER ultrastructures in humn embryonic epithelial cells of colonic mucosa、renal tubule and hepatocytes and also their alterations in embryogenesis.Methods　Transmission electromicroscopy technique.Results　With the embryonic development, the ER increased in amount, became complex in structure and with its characteristic ER structures in different cells. Conclusion　The changes of ER structures are one of the characters during cell differentiation.

There are more prognostic factors affecting the patients with cerebral hemorrhage, including bleeding volume, bleeding site, cause of bleeding, and blood pressure regulation during acute phase. Among them, the regulation of blood pressure during acute phase is a man-controlled important factor. However, the ideal blood pressure level has not yet to he determined at present. Therefore, more clinical trials are needed to determine the exact time window of blood pressure management in patients with intracerebral hemorrhage during acute phase and the range of blood pressure control.

The structure of social network and its information spreading characteristics in 41 users of Sina microb-logs were studied , which showed that the exchange level was quite low in the 41 users of health information in Sina microblogs, usually in small groups.Although the 41 users were found in the core area, their exchange was sporadic even with no exchange in the core nodes or in other nodes, thus effective information sharing could not be achieved and information could not be widely spread .

AIM:To develop a new method to determine cucurbitacin B in Pedicellus Melo by capillary zone(electrophoresis(CZE).) METHODS:All the samples were analysed on a fused silica capillary(75 cm?75 ?m I.D.) by using 50 mmol/L sodium borate solution(containing 5% acetonitrile) as the background electrolyte with the running voltage at 12.0 kV and detection wavelength of 265 nm.And clorprenaline hydrochloride was applied as the internal standard. RESULTS:The results showed a good linear correlation between relative peak area of cucurbitacin B(y) and its concentration(x) over the range of 0.15～1.2 mg/mL.Regression equation was y=(0.003 4x)+0.058,r=0.999 7,and the average recovery for baicalin was 100.2% with the RSD at(2.11%). CONCLUSION:The method is rapid,simple,reproducible and produces low pollution.It serves as a novel way to determine cucurbitacin B.

OBJECTIVE:To observe the clinical efficacy and safety of triple therapy of aspirin,clopidogrel and urinary kallidi-nogenase in the treatment of recurrent transient ischemic attack. METHODS:180 patients with recurrent transient ischemic attack were randomly divided into control group and observation group. Control group was orally treated with Aspirin enteric-coated tab-lets 100 mg,once a day + Clopidogrel hydrogen sulfate tablets 75 mg,once a day. Based on the treatment of control group,obser-vation group was additionally treated with Urinary kallidinogenase for injection 0.15 PNA unit adding into Sodium chloride injec-tion 100 ml by intravenous injection,once a day. The treatment course for both groups was 2 weeks. The clinic data was observed, including clinical efficacy,and LDL,HDL TC and TG levels before and after treatment,recurrence rate of cerebral ischemia,inci-dence of cerebral infarction and adverse reactions after 6 months of follow-up. RESULTS:The total effective rate in observation group was significantly higher than control group,and the recurrence rate of cerebral ischemia and incidence of cerebral infarction were significantly lower than control group(P<0.05). After treatment,HDL level in 2 groups were significantly higher than be-fore,and observation group was higher than control group;levels of LDL,TC and TG were significantly lower than before,and observation group was lower than control group(P<0.05). There were no severe adverse reactions in groups during treatment. CON-CLUSIONS:Triple therapy of aspirin,clopidogrel and urinary kallidinogenase has significant efficacy in the treatment of transient ischemic attack,with good safety.

Objective To investigate the influence of menstrucal cycle and anatomic site on the fractional anisotropy (FA) values of diffusion tensor imaging (DTI) in normal breast. Methods Prospectively enrolled 96 volunteers, who have identified with normal menstrucal phase and without breast diseases were found via the breast examination, ultrasound and MRI scan. The cases were divided into three groups according to menstrucal phase: menstrual period group(menstrual cramps 1 to 6 d), proliferative phase group(menstrual cramps 7 to 14 d) and secretory phase group(menstrual cramps 15 d to the next), and each group consisted of 32 subjects. All subjects were performed bilateral breast cross-sectional T1WI, T2WI, DWI and DTI scaning. On the nipple level figture, the mammary gland was divided into three regions including the anterior, central and posterior parts, and the FA values of the different phases and regions were measured. The Kruskal-Wallis H test was applied to analyse the difference of FA values in different menstrual phase and anatomic site. Results The FA values of the anterior region in menstrual phase, proliferative phase and secretary phase were 0.21 ± 0.07, 0.24 ± 0.09 and 0.17 ± 0.07, and the difference had significant difference(P=0.014).The FA values of the central region were respectively 0.15±0.08, 0.18±0.09 and 0.15±0.07, and without the statistically significant difference(P=0.090). The FA values of the posterior region were 0.21 ± 0.11, 0.24 ± 0.13 and 0.16 ± 0.11, and also showed significant difference(P=0.002). In different regions, the difference of FA values between menstrual phases and proliferative phases were also had statistically significant(P=0.018, 0.045, respectively). In the same region, the FA value was lowest in the secretary phase, and the proliferative phase was slightly higher than menstrual phase. Conclusion The FA values are affected by menstrual cycle and anatomic site.

The effect of cognition need on information seeking behaviors of information users and the difference in behaviors of information users were investigated in 184 undergraduates from the same grade with different back-grounds of subjects using the cognition need scale and undergraduates information-seeking behaviors scale ,respec-tively.The data were analyzed by descriptive statistical analysis, correlation analysis, rank test, and multivariate linear regression analysis , respectively , showing that the cognition need is significantly correlated with the informa-tion seeking behaviors of undergraduates in terms of information evaluation, information searching, information management and information use .

Objective　To prevent the after cataract induced by lens epithelial cells proliferation from postoperative cataract, monoclonal antibody (McAb) specific for rabbit lens epithelial cell is made, it will be the carrier further to be conjugated with cytotoxin. The conjugations will inhibit lens epithelial cells growth and not damage the other tissues of eye. Thereby McAb is the experimental bases of preventing after cataract.Methods　BALB/c mice were immunized by mixture of rabbit lens epithelial cells and Freund's adjuant. The immunized mouse spleen cells were fused with parental mouse myeloma cells (BALB/c　SP2/0) using polyethylene glycol (PEG-4000). The fused cells were selectively cultured by hypoxanthine, aminopterin and thymidine (HAT) culture medium. The specificities of the supernatant from hybridomas were tested by indirect immunofluorescence and immunohistochemistry SP (Streptavidin peroxdase conjugated method). The positive hybridomas were further cloned three times by methylcellulose culture medium to ensure monoclonality. At last, the consensual reaction of McAb was tested on human eye tissues.Results　Hybridomas were produced by fusion of spleen cells of immunized mice and mouse myoloma cells (SP2/0) with PEG-4000, and grown selectively in medium containing HAT after 16 days. Antibodies of the supernatant from hybridomas were tested on frozen sections of rabbit lens epithelial cell by indirect immunofluorescence. Only a positive clone secreted McAb against antigen of rabbit lens epithelial cell. The specificity of McAb was tested on paraffin sections of whole rabbit eye and whole human eye by immunohistochemistry SP. The results indicated that McAb was only positive to rabbit lens epithelial cell membrane and it was negative to the other tissues of rabbit or whole human eye tissues.Conclusions　McAb specific for rabbit lens epithelial cell was manufactured successfully. The specificity of McAb was strong. There was no consensual reaction on human eye tissues. It might be the experimental bases of further targeting chemotherapy on after cataract.