Lipase gene from Penicillium expansum(lip07) was cloned and over-expressed in Pichia pastoris.a random mutant named ep8,which contained a single amino acid substitution,was obtained by using the lip07 as an error-prone PCR template in previous study.ep8 shows higher thermostability than that of lip07,To further improve the thermostability of the lipase,the Lys of wild-type(lip07) and mutant(ep8) in 202 were substituted by Ala using the Overlap extension PCR technique respectively.The mutant genes(lip07-K202A and ep8-K202A) were subcloned into pAO815,and then transformed into the Pichia pastoris GS115 for extracelluar expression,respectively.15% SDS-PAGE analysis indicated that the molecular mass of PEL-ep8-K202A and PEL-lip07-K202A are both about 28kDa,which is same with the wild-type lipase.The Tm of PEL-ep8-K202A is 41.66℃,2.63℃ higher than that of the wild-type(39.03℃) and 1.21℃ higher than the random mutant(PEL-ep8:40.45℃);the Tm of single mutant(PEL-lip07-K202A) is 37.08℃,2℃ lower than that of the wild-type lipase.

The process of U(VI) biosorption by freshwater algae Chlorella pyrenoidosa and its absorption mechanism, absorption thermodynamics and absorption kinetics were investigated in this paper. The effects of pH, contact time, initial U(VI) concentration and temperature on biosorption were studied respectively. Research result showed that the absorption effect of U(VI) by Chlorella pyrenoidosa was affected by pH value of solution to a great extent, the absorption reached its balance within 5 min with optimal pH value 6 and max absorption quantity 2.7 mg/g. On the other hand, the absorption quantity of U(VI) by Chlorella pyrenoidosa was positively correlated with the initial concentration of U(VI); and the absorption quantity did not fluctuate remarkably when temperature was varied at the range of 20℃ to 30℃. Research result also showed that the process of U(VI) absorption was congruent with the second order kinetic model, and the correlation coefficient was high reaching to 0.99. It was suggested that the U(VI) biosorption by Chlorella pyrenoidosa was a complicated process consisting of many simultaneous reactions and could be described by Languir model quite well.

A high productive poly ?-glutamic acid(?-PGA) strain PGA-N in a culture medium containing no L-glutamine was isolated from fermentation products.With the following identifications of colony mor-phology,physiological and biochemistry experiments,and genetics,the strain PGA-N was classified as a Bacillus licheniformis.According to the product environment,the base culture medium having no L-glutamine was simulated.In order to enhance the production of the strain PGA-N,the fermentation condi-tions,such as carbon source,nitrogen source,were optimized and the ?-glutamic acid production reached 5.16 g/L after getting the optimum formulation of this culture medium.PGA-N was mutagenized with com-bination of NTG and UV.A mutant PGA-N-C10 was screened which PGA production was increased from 5.16 g/L to 8.82 g/L.The study also investigated the effects of agitation speed on the cell biomass,?-PGA production and the ?-PGA molecular weight.The ?-PGA yield of PGA-N-C10 was as high as 11.00 g/L when the agitation speed was 400 r/min.

Objective To explore the relationship between Wuyishan cliff tea and esophageal cancer,and make integrated measures to prevent and control esophageal cancer.Methods A case-control study was conducted with a ratio of 1 to 1 and a health survey was conducted on objects which included 92 cases with histologically confirmed diagnoses of squamous-cell carcinoma of the esophagus and 92 controls matched by age,sex and resident.The analysis method of conditional logistic regression was used to analyze the influence of cliff tea on the development of esophageal cancer. Results The frequency of cliff tea drinking was 73.91% in cases with esophageal cancer compared with 84.78% in controls. As compared with subjects without tea drinking,subjects with cliff tea drinking significantly decreased the risk for developing esophageal cancer (P<0.05),and its OR were 0.60.The risk of esophageal cancer significantly reduced with increasing frequency of cliff tea drinking per week. Furthermore, we also found that the risk of esophageal cancer significantly reduced with the duration of cliff tea drinking and the early of beginning to drink cliff tea.Conclusion Wuyishan cliff tea drinking may protect against the risk of esophageal cancer.

BACKGROUND: It has been proved that platelet activation is involved in the development of diabetic angiopathy. Glycoprotein (GP) Ib/IX/V complex is one of the main platelet membrane glycoproteins, and the receptor of both von Willebrand Factor and thrombin. It plays a key role in the process of platelet activation.OBJECTIVE: To observe the expression changes of GP Ib/IX/V complex and its component GP Ibα in patients with type 2 diabetes mellitus. DESIGN: Case-control study. SETTING: Department of Integrated Chinese and Western Medicine, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology.PARTICIPANTS: A total of 51 type 2 diabetic outpatients who visited Union Hospital were enrolled from December 2005 to January 2007. The diagnosis was based on the independent criteria from WHO in 1999. Of all the 51 patients, there were 23 females and 28 males, with a peripheral platelet count of over 50×109 L-1. All the subjects had no history of administrating drugs two weeks before the examination, which would potentially influence platelet count. According to disease controlling condition, the patients were assigned to well controlled patient (WCP) group (n = 25) and poorly controlled patient (PCP) group (n =26); and according to whether angiopathy was accompanied, diabetic patients were divided into vascular disease (VD) group (n =27) and non-vascular disease (NVD) group (n =24). Meanwhile 23 healthy subjects were enrolled as normal control group. Informed consents were obtained from subjects and their relatives. The experiments were approved by the ethical committee of Union Hospital.METHODS: Fasting venous blood was harvested from all the subjects' elbow on the early morning of visit day.①Biochemical analysis: Fasting plasma glucose (FPG), glycosylated hemoglobin (HbA1c) and fasting plasma insulin (FINS) was measured by the Clinical Chemistry Department in Union Hospital.② Measurement of platelet membrane GP Ib/IX/V complex and its component GP Ibα: First, 3 mL cubital fasting blood was drawn from each subject and was anti-coagulated with 38 g/L natrium citricum. After that, all samples were fixed with 10 g/L paraform for 45 minutes. Then 50 μL well-fixed blood was added into the polystyrene tube, meanwhile 20 μL monoclonal antibody, such as CD42a-b-c-d and PE-labeled CD42b, was respectively mixed gently with the blood sample and incubated at room temperature in dark for 30 minutes. Next, 20 μL FITC-labeled rat IgG was mixed with the sample containing CD42a-b-c-d and incubated equally. In the end all blood samples were analyzed by FACS420 flow cytometry and the results were expressed as mean fluorescence intensity (MFI). ③ Platelet maximun aggregation rate (MAR) was detected according to reference.MAIN OUTCOME MEASURES: ①Biochemical indicators;②The expressions of GP Ib/IX/V complex and GP Ibα;③Platelet MAR.RESULTS: Fifty-one patients with type 2 diabetes mellitus and twenty-three healthy subjects were all involved in the result analysis.①There were significant differences in FINS in WCP group and PCP group compared with normal controls (P ＜ 0.01). FPG and HbA1c were significantly higher in PCP group compared with normal control group and WCP group (P ＜ 0.01).②Expressions of GP Ib/IX/V complex and GP Ibα were significantly lower in WCP group and PCP group compared with normal control group (P ＜ 0.01), significantly lower in PCP group than in WCP group (P ＜ 0.05), and also significantly lower in VD group and NVD group compared with normal control group (P ＜ 0.01). Moreover the expression of GP Ibα in VD group and NVD group was significantly lower than that of normal control group (P ＜ 0.05), and it also significantly decreased in VD group compared with NVD group (P ＜ 0.05). MFI of GP Ib/IX/V complex had an obvious negative correlation with FBG, HbA1c and FINS (r =-0.634, -0.573, -0.649, P ＜ 0.05), and GP Ibα MFI was obviously negatively correlated with FBG and HbA1c (r =-0.602, -0.543, P ＜ 0.05).③Platelet MAR of diabetic patients were remarkably higher than in healthy subjects (t =-3.852, P ＜ 0.01). Platelet MAR in PCP and VD groups were respectively higher than those in WCP and NVD groups (P ＜ 0.05). CONCLUSION: Platelet activation exists in the early stage of type 2 diabetes mellitus without diabetic angiopathy, and is more obvious after diabetic angiopathy. There is a positive correlation between platelet activation and blood glucose. As a receptor of thrombin and von Willebrand Factor, GP Ib/IX/V complex may be involved in the development of diabetic angiopathy.

OBJECTIVETo explore potency material bases of Xuebijing (XBJ) formula, and to analyze its effects at the molecular network level.

METHODSTotally 16 sepsis-related targets were selected and classified into three categories such as inflammation, immune, and coagulation referring to biological roles. Then molecular database of chemical compositions in XBJ formula were constructed to explore mutual actions with inflammation, immune, and coagulation targets.

RESULTSDanshen root and safflower, with more effector molecules with immune and coagulation targets, have extensive anticoagulation and anti-inflammation effects. The former 10 molecules with better mutual actions with sepsis targets were sequenced as tryptophane, danshensu, gallic acid, salvianolic acid D, protocatechuic acid, salvianolic acid A, danshensu C, vanillic acid, rosmarinic acid, phenylalanine. There existed two phenomena in XBJ formula as follows. One component had stronger actions with multi-targets, for example, danshensu had actions with 13 targets. Meanwhile, different components acted on the same target protein, for example, 8 molecules acted with MD-2.

CONCLUSIONXBJ formula had certain potential synergistic effects with sepsis targets, which could provide certain referential roles for findina new type anti-septic drugs.

Caffeic Acids ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; therapeutic use ; Gallic Acid ; Hydroxybenzoates ; Inflammation ; Lactates ; Sepsis ; drug therapy

Humans ; Liver Diseases ; Practice Guidelines as Topic ; Prognosis

Purpose:To further testify the role of costimulatory molecular B7 in renal ischemia/reperfusion in-jury. Methods:The mRNA transcription level of B7 was assayed with reverse transcription semiquantative poly-merase chain reaction in the uninephrectomized rat model established by warm ischemia/reperfusion. Results :ThemRNA transcription level of B7 was very low in normal and ischemia renal tissue, but was gradually increased af-ter repefusion,and reached peak level at 72 hours after reperfusion; The mRNA transription level of B7 in the is-chemia 60 min reperfusion group was higher than that in the ischemia 30 min reperfusion group. Conclusions :Up-regulation of costimulatory molecular B7 further elucidates the mechanism of relationship between ischemia/reperfusion injury and acute rejection; More heavier ischemia/reperfusion injury, more possible acute rejection.

A poly-?-glutamic acid producing strain--BLN-2, was isolated from the soybean products. According to the biochemical characteristics and 16S rRNA, the strain was identified as Bacillus subtilis. Using soybeans as culture, the solid-state fermentation conditions of BLN-2 have been studied. The results showed that the optimal carbon and nitrogen sources of BLN-2 were glucose, fructose, NaNO3 and KNO3, respectively. The orthrogonal experiments showed, when the final concentration of the fructose which was added to the soybean culture was 0.5%, the glucose, NaNO3 and KNO3 final concentraion were 2.0%, the production of ?-PGA was the highest--89.05 g/kg. It is 48.42% higher than other comparable soybean medium under the same conditions.

BACKGROUNDThere are few effective methods for treating injuries to the lower trunk of brachial plexus, and the curative effect is usually poor. The purpose of this study was to provide anatomic references for transferring the brachialis muscle branch of musculocutaneous nerve (BMBMCN) for selective neurotization of finger flexion in brachial plexus lower trunk injury, and to evaluate its clinical curative effects.

METHODSMicroanatomy and measurement were done on 50 limbs from 25 adult human cadavers to observe the origin, branch, type of the BMBMCN and median nerve, as well as their adjacent structures. Internal topographic features of the fascicular groups of the median nerve at the level of the BMBMCN were observed. In addition, the technique of BMBMCN transfer for selective neurotization of finger flexion of the median nerve was designed and tested in 6 fresh adult human cadavers. Acetylcholinesterase (AchE) staining of the BMBMCN and median nerve was done to observe the features of the nerve fibers. This technique was clinically tried to restore digital flexion in 6 cases of adult brachial plexus lower trunk injury. These cases were followed up for 3, 6, 9 and 12 months postoperatively. Recovery of function, grip strength, nerve electrophysiology and muscle power of the affected limbs were observed and measured.

RESULTSThe brachialis muscle was totally innervated by the musculocutaneous nerve (MCN). Based on the Hunter's line, the level of the origin of the BMBMCN was (13.18 +/- 2.77) cm. AchE histochemical staining indicated that the BMBMCN were totally made up of medullated nerve fibers. At the level of the BMBMCN, the median nerve consistently collected into three fascicular groups as shown by microanatomy in combination with AchE stain. The posterior fascicular group was mainly composed of anterior interosseous nerves and branches to the palmaris longus. The technique was tested in six fresh cadavers successfully, except that stoma split occurred in one case. Five of the six cases recovered digital flexion 12 months after operation, and at the same time grip strength, muscle power, and nerve electrophysiology also recovered markedly.

CONCLUSIONSThe technique of transferring the BMBMCN for selective neurotization of finger flexion is anatomically safe and effective, with satisfactory clinical outcomes.

Acetylcholinesterase ; analysis ; Adult ; Brachial Plexus ; anatomy & histology ; injuries ; Brachial Plexus Neuropathies ; surgery ; Clinical Trials as Topic ; Female ; Humans ; Male ; Middle Aged ; Musculocutaneous Nerve ; transplantation ; Nerve Transfer ; methods ; Retrospective Studies