Based of the study on the impact of protoplasts under different conditions of enzymolysis solution, culture time, enzymolysis time and enzymolysis temperature in sequence using orthogonal test. The best optimal condition of producing protoplast was :0.5% helicase add 0.5% lysozyme and 1.0% cellulase, enzymolysis time was 3 hours , enzymolysis temperature was 32℃,under these condition the production of protoplast can be 4.25?10~(6 )per mL. Ca~(2+) and PEG can increase the regeneration ratio of protoplast.After induced mutation by complex factor under DES,UV and LiCl,the mutagenesis strain which own increasing enzyme activity and transmissibility steaty was obtained.

Ras is a kind of protein closely related to neoplasms,the persistent activation of which would induce tumorigenesis and progression.RASAL1 is a Ca~(2+) regulated ras GTPase-activating-like protein, the loss or decreased expression of RASAL1 induces ras activation, then results in tumongenesis and progression.In this review we sum up the mechanism of ras activation, the relationship between neoplasms and RASAL1 and the mechanism of RASAL1 expression decreasing.

Objective: To find the reason of risk on medical equipment. Methods: Based on RCA, to adopt the event classification to evaluate the X-ray equipment, high-frequency electro tome, electro-cardiac monitor, infusion pump, ventilator, defibrillators and infant incubator, discuss the root cause of medical risk. Results:To deduce the medical equipment risk by PM and quality control. Conclusion:With the analysis of quality control, the application of RCA is gradually explained in management of medical equipment.

OBJECTIVE To investigate the infection and drug-resistance of Escherichia coli.METHODS Double disk diffusion test and improved three-dimensional test were adopted to test ESBLs and AmpC of 135 E.coli strains obtained from bile cultivation from operations of biliary inpatients in our hospital from 2003 to 2005,and King-Bauer′s was used to test the sensitivity of 14 kinds of common antibiotics.RESULTS The detection rate of ESBLs producing bacteria was 30.4%(41 strains),and that of AmpC producing bacteria was 5.2%(7 strains);drug-sensitivity test indicated,that E.coli was sensitive to meropenem,imipenem,cefoperazone / subactam,ciprofloxacin and amikacin.CONCLUSIONS E.coli is a main bacterium in biliary infection,and the strains producing ESBLs and /or AmpC are resistant to the cephalosporins of third and fourth generation;prematurely and excessively using broad-spectrum antibiotics before operation is the primary cause to increase in enzyme-producing strains.

Objective To assess the effects of hydrocortisone (HC) on brain injury in septic shock rats induced by lipopolysaccharide (LPS).Methods Pathogen-free Wistar male rats were chosen to produce septic shock model by injection of LPS (Escherichia coil,O55:B5),25 mg/kg LPS was given by femoral vein.All rats in accordance with the random number table,were randomly divided into 3 groups:normal control group,LPS group,and low dose of HC group (HC 5 mg/kg).Every group had ten rats.Six hours after the septic shock model was established,the blood sample and brain tissue were collected to perform histopathological examination and measure the serum level of neuron specific enolase (NSE).Results The serum level of NSE,brain water content,inflammation score,neural damage score,vascular reaction score in LPS group were significantly higher than those in control group [(10.078 ± 0.526) μg/L vs.(3.719 ± 0.602) μg/L,(82.328 ±0.011) ％ vs.(78.896 ±0.005) ％,(2.750 ±0.463) scores vs.(0.125 ±0.345) scores,(2.875 ± 0.353) scores vs.(1.125 ± 0.354) scores,(2.875 ± 0.463) scores vs.(0.500 ± 0.534) scores,P ＜ 0.01,respectively].After HC treatment,the serum level of NSE,brain water content,and brain tissue pathological score [(6.297 ± 1.393) μg/L,(80.283 ± 0.005) ％,(0.625 ± 0.744) scores,(1.375 ± 0.744) scores,(1.250 ± 0.463) scores] were significantly lower than those in LPS group (P ＜ 0.05,respectively).Conclusion After giving low dose of HC,the serum level of NSE,brain edema,brain pathological changes all got improvement,which indicated that low dose HC could have protect effect on septic encephalopathy induced by LPS.

Electrical status epilepticus in sleep (ESES) indicates a special electroencephalographic pattern showing sleep-induced continuous paroxysmal discharges of spike-wave complexes.ESES can be seen in a series of epileptic syndromes in children characterized by seizures,ESES and cognitive impairment,including epilepsy with continuous spikes and waves during slow sleep,Landau Kleffner syndrome,typical and atypical benign childhood epilepsy with centro-temporal spikes.Though the mechanism generating ESES remains elusive,great advances in several aspects,such as genetic studies (GRIN2A,copy number variations,ect.),high-frequency oscillations and brain networks have been achieved.Treatment for ESES related disorders should focus on both epileptic seizures and ESES,and the introduction of appropriate antiepileptic drugs or other strategies like hormone therapy should be considered to furthest eliminate epileptic seizures and protect cognitive function.

In our university,the training for geriatric postgraduates of clinical medicine is now in the initial stage.Therefore,it is important to explore the training model to improve the quality of postgraduate.The program focused on cultivating geriatric postgraduates' ability of diagnosis and treatment,helping them have the concept of integral service and enhancing their teaching and thinking ability of clinical sciences to provide high-level comprehensive geriatric doctors for the society.

Fusion PCR, which employs chimeric primers to generate PCR products with complementary ends in its amplifications, is a rapid and flexible method in joining different DNA fragments. This method can assemble DNA fragments without the treatment of restriction endonucleases and T4 DNA ligase. It offered a shortcut for the construction of homologous recombinant fragments. Through assembling three recombinant fragments successfully, fusion PCR procedure was improved and manipulation essentials of fusion PCR were described in detail. The results indicated that the improved procedure can assemble three or four fragments simultaneously, and the length of each fusion product is above 4.5 kb. The result recombinants were proposed to be use in further experiments, which structures had been confirmed by sequencing.

Based on a ?-tubulin gene EST sequence from Trichoderma harzianum mycelium cDNA library,a 1.74kb ?-tubulin gene coding region,together with a 1.5kb 5'-flanking region and a 1.0kb 3'-flanking region was cloned from T.harzianum by IPCR methods.The gene encodes a 446 amino acid polypeptide,which shows a high degree of homology with other fungal ?-tubulins.A three-dimensional model of the T.harzianum ?-tubulin was built based on the crystal structure of bovine tubulin using homology modeling method.Tertiary protein structure of the ?-tubulin should provide a basis for understanding a significant body of research on tubulin's properties and interactions with antimicrotubule agents.

Objective To investigate the clinical effect of insulin on gestational diabetes mellitus(GDM).Methods GDM patients were randomly divided into group A and group B,group B were treated with short term intensive insulin pump therapy,and group A were treated with insulin pump in short term combined with biosynthetic human insulin therapy.Results The body mass index and diabetes family history rates pre-pregnancy in group B were significantly higher than group A(P<0.05).Logistic analysis showed that family history of BMI pre-pregnancy and diabetes family history were independent risk factors for GDM drug treatment(P<0.05).Compared with the group B,the improvement of blood glucose levels and the incidence of complications in patients with group A were better than the former(P<0.05).Conclusion Rational application of insulin can improve the efficacy of clinical treatment of GDM, reduce the incidence of complications, with high safety.