The aim of the work is to improve the synthetic process of dexrabeprazole sodium,enhance quality and yield of the product,simplify synthetic steps,and offer a stable and feasible process. Starting from 2-[[[4-(3-methoxypropoxy)-3-methylpyridine-2-yl]methyl]thio]-1 H-benzimidazole,dexrabeprazole was produced by asym-metric oxidation reaction with oxidant cumene hydroperoxide in the presence of chiral catalyst tetraisopropyl titan-ate and L-(+)-tartaric acid diethyl ester. Dexrabeprazole sodium was obtained by the reaction of purified dexrabeprazole with sodium hydroxide in a total yield of 79%with an HPLC purity of ＞99. 5%. The structure of dexrabeprazole sodium was confirmed by NMR,IR,elemental analysis and LC-MS. The improved process of dexrabeprazole sodium possesses simple operations,good yield and high purity,which is feasible for industrializa-tion.

In vitro evolution methods are often used to modify protein with improved characteristics. We developed a directed evolution protocol to enhance the thermostability of the beta-1,3-1,4-glucanase. The thermostability of the enzyme was significantly improved after two rounds of directed evolution. Three variants with higher thermostability were obtained. The mutant enzymes were further analyzed by their melting temperature, halftime and kinetic parameters. Comparing to intact enzyme, the T50 of mutant enzymes 2-JF-01, 2-JF-02 and 2-JF-03 were increased by 2.2 degrees C, 5.5 degrees C and 3.5 degrees C, respectively, the halftime (t1/2, 60 degrees C) of mutant enzymes 2-JF-01, 2-JF-02 and 2-JF-03 were shortened by 4,13 and 17 min, respectively, the V(max) of mutant enzymes were decreased by 8.3%, 2.6% and 10.6%, respectively, while K(m) of mutant enzymes were nearly unchanged. Sequence analysis revealed seven single amino acid mutant happened among three mutant enzymes, such as 2-JF-01 (N36S, G213R), 2-JF-02 (C86R, S115I, N150G) and 2-JF-03 (E156V, K105R). Homology-modeling showed that five of seven substituted amino acids were located on the surface of or in hole of protein. 42.8% of substituted amino acids were arginine, which indicated that arginine may play a role in the improvement of the thermostability of the beta-1,3-1,4-glucanase.This study provide some intresting results of the structural basis of the thermostability of beta-1,3-1,4-glucanase,and provide some new point of view in modifying enzyme for future industrial use.
Amino Acid Sequence ; Bacillus ; enzymology ; Directed Molecular Evolution ; Endo-1,3(4)-beta-Glucanase ; biosynthesis ; chemistry ; genetics ; Enzyme Stability ; Hot Temperature ; Molecular Sequence Data ; Recombinant Fusion Proteins ; biosynthesis ; chemistry ; genetics

To compare of performance of beta-1,3-1,4-glucanase gene (bgl) in different expression systems, the beta-1,3-1,4-glucanase gene (GenBank Accession No. EU623974) was amplified from Bacillus amyloliquefaciens BS5582 by PCR and was cloned into three vectors pEGX-4T-1, pET20b(+) and pET28a(+) to construct pEGX-4T-1-bgl, pET20b(+)-bgl and pET28a(+)-bgl recombinant plasmids. The pEGX-4T-1-bgl was transformed into three different Escherichia coli host strains. The pET20b (+)-bgl and pET28a (+)-bgl were transformed into E. coli BL21 (DE3) respectively. Recombinant beta-glucanase was expressed by IPTG inducement in these recombinants. E. coli BL21 (DE3)-pET28a (+)-bgl had the highest enzyme activity. In Luria-Bertani medium, the total enzyme activity was (322.0 +/- 8.8) U/mL, which was 40.1% of original strain in optimal shaking flask condition. This recombinant's performance was studied in Terrific Broth medium under inducement of IPTG and lactose at the same time., and the highest total enzyme activity could reach (1883.3 +/- 45.8) U/mL (818.8% of the original), which indicate that the recombinant strain has a good value in industry application.
Bacillus ; enzymology ; genetics ; Cloning, Molecular ; Endo-1,3(4)-beta-Glucanase ; genetics ; metabolism ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; Molecular Sequence Data ; Recombinant Proteins ; genetics ; metabolism

The ECM25 deletion mutant of industrial brewing yeast, G03/a, was constructed by replacing the ECM25 gene with the kanMX gene. The transformant was verified to be genetically stable. The PCR analysis showed that ECM25 gene in the G-03/a was deleted. Under aerobic conditions of ll degrees C and 28 degrees C, compared with the host strain G-03, the excretive glutathione concentration of G-03/a increased by 21.4% and 14.7%, respectively. Strains G-03 and G-03/a were inoculated in flasks and cultivated continuously for 4 generations. Compared with the host strain G-03, the glutathione concentration in the main fermentation broth and final beer of strain G-03/a increased by 32.1% and 13.8%, the stability index (SI) increased by 7.7% and 5.3%, respectively, and the flavor resistance staling value (RSV value) in final beer increased by 45.0%. During EBC fermentation, the glutathione concentration in the main fermentation broth of strain G-03/a increased by 34.0%, compared with the host strain G-03. Furthermore, no significant difference in routine fermentation parameters was found. The strain G-03/a is proved to be an excellent anti-staling brewing yeast to improve beer flavor stability.
Beer ; microbiology ; Fermentation ; Gene Knockout Techniques ; Glutathione ; secretion ; Recombination, Genetic ; Saccharomyces ; genetics

BACKGROUND: Researches on the hand allograft have been transited from laboratory aspects to clinical application.OBJECTIVE: To evaluate the clinical significance of serial monitoring of co-stimulating signals in double hand transplantation.DESIGN: Case controlled observation.SETTING: Department of Traumatic Orthopaedics, Nanfang Hospital,Southern Medical University.PARTICIPANTS: From September 2001 to December 2001, a patient who was proposed to have double hand transplantation in the Department of Traumatic Orthopaedics, Nanfang Hospital Affiliated to Southern Medical University was taken as the subject of the experiment, meanwhile 15healthy adult man were enrolled as controls.METHODS: Before operation, blood sample was collected from the patient once before given immunodepressant and twice after given immunodepressant; blood sample was also collected before arteriovenous connection during operation, as well as once a day during the first week after operation, once every other day on the second week, twice a week on the third and fourth week, followed by once a week for total two month. The first week after operation was taken as inducing period with the remainder as maintaining period. 2 mL peripheral venous blood was collected from healthy adult and anti-coagulated by EDTA of mass scores of 20 g/L.Flowcytometry was used to carry out a serial monitoring of T cell surface co-stimulating moleculars (CD28, CD54, CD11a).MAIN OUTCOME MEASURES: The change of co-stimulatingmolecules on the surface of T lymphocyte before and after transplantation.RESULTS:The changes of the three co-stimulating molecules were consistent with each other after double hand transplantation (CD28,CD54,CD11a),displaying decrement during inducing period comparing to pre-operation [(9.84±5.28)%, (55.50±3.62)%; (71.03±5.33)%, (95.10±1.26)%;(9.40±9.17)%, (29.70±3.23)%] and keeping at lower level at maintaining period [ (22.54±6.56) %, (91.28±8.12 ) %, ( 11.22±4.08 ) %].CONCLUSION:The serial monitoring of the co-stimulating signals in the peripheral blood of patients with linb allograft is beneficial to the observation of the posopertaive immunologic reaction, providing important guidance for the application of immunosuppression medicine, as well as the prediction and diagnosis of postoperative immune rejection. Meanwhile, costimulating signal induced immunological tolerance is proved promising for the application of hand transplantation.

While the basic concept of saving life and decreasing disability h as not changed in the management of polytrauma, the timing and strategies of tre atment have been gradually modified. Damage control is one of the new strategies , and its application in orthopedic traumatology is known as damage control orth opedics. Damage control orthopedics involves formalized three stages in the mana gement of fractures of long bones and pelvis. Stage 1 consists of temporary fixa tion of the unstable fractures and hemostasis. Stage 2 comprises resuscitation, warming, oxygen delivery and administration of coagulation factors to the patien t in the intensive care unit. In Stage 3 definitive fixation is done for the fra cture. Indications of damage control orthopedics: for patients with polytrauma w hose conditions are unstable or in extremis the damage control strategy is recom mended. For the borderline patients early total care may be applied, but the sur gery should be performed with great caution. It has to be converted to the damag e control strategy if conditions of the patient deteriorate during the operation . Since damage control orthopedics is an evolving practice, further work is need ed to enhance its effectiveness and to reduce the incidence of ARDS and MOF.

Modern Chinese Orthopaedic traumatology has stemmed from the orthopaed ics of Traditional Chinese Medicine(TMC)and western orthopaedics.On the one hand,with a long history and its unique system of theory and treatment,TMC orthopaedics h as made a great con-tribution to the development of modern Chinese orthopaedic traumatology.On the other hand,the introduction and development of western medicine have dramatically pro moted the development of modern Chinese orthopaedic traumatology.Modern Chinese surgeons of orthopaedic trauma have accomplished numerous successful f irst cases in the world since 1950s an d have thus expanded their professional influence.At pr esent,the Chinese orthopaedic trau ma community has grown much stronger and established its ow n academic associations and journals.More and more de-partments of orthopaedic traumatology have been successively establis hed as an independent en-tity in large and middle-sized hospi tals in China.So far,the Chinese orthopaedic trauma com-munity,on the whole,has reached the international level,but there are q uite a lot to be improved in the aspect of treatment.To end the backwardness,in the future Chinese orthopaedic trauma community should make great efforts in education of orthopedists on comp rehensive knowledge,establishment of formal profession al training system,national standardization of orthopaedic procedures in clinical diagnosis an d treatment,active membership of re lative international aca-demic associations and increasing p rofessional exchanges with international counterparts,as well as in research and development of operative techniques and fixators.All these will be the main strategies and developmental trends in the future for Chinese orthopaed ic traumatology. [

OBJECTIVETo discuss the diagnosis and operative technology of acetabular fracture.

METHODS41 cases of displaced acetabular fractures were enrolled. CT and 3D-CT were performed before operation to understand the classification of fractures and to determine operative regimen. All cases were fixed by reconstruction plates and screws through the best approach.

RESULTS39 cases were followed up for 1 - 4 years, with an average of 1.5 years. Curative outcomes were assessed by Matta's criteria. Clinically, 16 (53.3%) 30 fresh fractures were excellent, 11 (36.7%) good, 3 (10%) fair. The rate of excellence and goodness was 90%. In 9 cases of old fractures, 2 were excellent, 3 good, 2 fair and 2 poor. The rate of excellence and goodness was 55.6%. Complications consisted of sciatic never injury, necrosis of the femoral head, and heterotopic ossification.

CONCLUSIONSThe time of operation, degree of injury, and quality of reduction of fracture are important factors affecting the postoperative function of the hip. The key to enhance the outcome is to verdict the type of fracture, select the proper approach before operation, and precise reduction and appropriate internal fixation during operation.

Acetabulum ; injuries ; Adolescent ; Adult ; Female ; Fracture Fixation, Internal ; methods ; Fractures, Bone ; surgery ; Humans ; Male ; Middle Aged

OBJECTIVETo study the psychology and the treatment during the process of hand allograft.

METHODSThe patients were interviewed to evaluate their states of mind and their abilities to manage stress during the selection of patients. The psychology of the two patients were trained before the operation and managed accordingly afterwards.

RESULTSOne of 12 candidates was found to be unsuitable for the transplantation because of psychiatric problems. One week postoperatively, the two patients were anxious, lacked patience and were afraid of seeing the long-awaited grafted hand. After 1 week, the patients began to accept the new hand, with full acceptance of the hand 1 month later. With the recovery of hand sensation 4 to 5 months after the operation, the patients regarded the hand as their own.

CONCLUSIONPsychological problems exist during preoperative selection of patients and postoperative rehabilitation, requiring psychologists in the hand transplantation team.

Adult ; Female ; Hand Transplantation ; Humans ; Psychotherapy ; Transplantation ; psychology ; Transplantation, Homologous

The major surgical managements for tibial plateau fractures are anatomical reduction, firm fixation and bone grafting. There have appeared many new techniques in recent years, such as less invasive stability system (LISS) and arthroscopy assisted treatment. But such complications as loss of reduction, infection, soft tissue lesion around the knee, nonunion, posttraumatic arthritis, and knee stiffness seem to be inevitable. This article introduces the mechanism, treatment and prevention of the complications of the tibial plateau fracture.