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.

Objectives To develop and evaluate a real-time fluorescent quantitative RT-PCR method for the detection of a novel CXC chemokine macrophage inflammatory protein-2?(MIP-2?)mRNA expression based on TaqMan technique.Method The expression of MIP-2? mRNA was detected by TaqMan real time fluorscet quantitative RT-PCR. Results The dynamic range of the assay varied from 102 to 107 copies. The intra- and inter-assay coefficient variation (CV) were less than

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.

Objective To investigate and analyze the effect of psychological intervention combined with paclitaxel and carboplatin on the clinical efficacy and quality of life of locally advanced cervical cancer. Methods 100 cases of locally advanced cervical cancer treated in our hospital from February 2015 to November 2016 were selected and randomly divided into the control group and the experimental group, with 50 patients in each group. The control group were treated with neoadjuvant chemotherapy with cisplatin combined with fluorouracil, experimental group were treated with paclitaxel and carboplatin chemotherapy, and give attention to patients with psychological intervention, mental health, strengthen communication and exchanges, the elimination of patients with anxiety. The treatment effect and quality of life index of 2 groups were compared and analyzed. Results After treatment, the total score of quality of life in the experimental group was (75.2 ± 11.2) points, significantly higher than that of the control group (64.2 ± 9.3), with statistical difference (P<0.05). Among the patients in the experimental group, 5 patients progressed, and 9 patients were stable. The number of effective treatment was 36. In the control group, 10 patients progressed, 13 patients were stable, and the number of effective treatment was 27. Conclusion The clinical effect of psychological intervention combined with paclitaxel and carboplatin for local advanced cervical cancer is better, higher efficiency, can significantly improve the quality of life of patients, eliminate the negative emotion of patients, which has clinical significance.

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.

Objective To investigate the clinical efficacy of acupuncture plus Jisheng Shenqi pills in treating patients with diabetic neurogenic bladder. Method Patients with diabetic neurogenic bladder were allocated, using a random number table, to treatment and control groups. The treatment group received acupuncture plus Jisheng Shenqi pills and the control group, oral administration of Western medicine. Both groups of patients were treated for four weeks as a course. Post-treatment clinical total efficacy rates, pre-treatment and post-treatment postvoid residual urine volumes, and adverse reactions during treatment were compared between the two groups. Result The total efficacy rate was higher in the treatment group (93.8%) than in the control group (68.8%) (P<0.05). The postvoid residual urine volume decreased significantly in both groups of patients after treatment compared with before (P<0.05). Post-treatment postvoid residual urine volume was lower in the treatment group than in the control group (P<0.05). The adverse reaction rate was lower in the treatment group than in the control group (P<0.05). Conclusion Acupuncture plus Jisheng Shenqi pills is remarkably effective in treating patients with diabetic neurogenic bladder. It can effectively decrease postvoid residual urine volume in the patients, has higher safety and is worthy of clinical popularization.

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.

As an important device of modern medical treatment,medical monitor is a combination of modern electronics technology,computer technology and medical technology.In this paper,the development of medical monitors in China is summarized,especially the main technique characteristics of medical monitor in Chinese market.The examples of monitors made in China are presented.

Objective To explore effect of miR‐302a on proliferation and apoptosis in folate deficiency mouse embryonic stem cell (mESC) .Methods The cases were divided into complete culture medium group (control group) ,folate‐deficient culture medi‐um group (folate‐deficient group) ,folate‐deficient culture medium plus miR‐302a mimic group (miR‐302a group) .The expression of miR‐302a was examed by RT‐PCR in control group and folate‐deficient group .To construct miR‐302a mimic and then was trans‐fected into the folate‐deficient culture medium mESC .Effect of miR‐302a mimic on mESC viability was detected by MTT assay ,the effect of miR‐302a mimic on mESC apoptosis was examed by Annexin V‐FITC/PI flow dual‐staining method ,the effect of miR‐302a mimic on mESC cycle was examed by flow cytometry .The activation of phosphatidylinositol 3 kinase (PI3K)/protein kinase B (AKT)/mammalian target of rapamycin (mTOR) and expression of CyclinD1 ,p21 and p27 was assayed by Western blot . Results The expression of miR‐302a was lower in folate‐deficient group(P<0 .01) .Compared with control group ,the viability of mESC was lower ,the apoptosis of mESC was higher ,the cell cycle was arrested in G1 phase ,the level of phosphorylation of AKT and mTOR was lower ,the expression of CyclinD1 was lower ,the expression of p21 and p27 was higher in folate‐deficient group , with statistical significance (P<0 .01) .Compared with folate‐deficient group ,the viability of mESC was higher ,the apoptosis of mESC was lower ,G1 phase was shortened ,the level of phosphorylation of AKT and mTOR was higher ,the expression of CyclinD1 was higher ,the expression of p21 and p27 was lower in miR‐302a group with statistical significance (P<0 .01) .Conclusion These results suggested miR‐302a exerted anti‐apoptosis and promote cell proliferation in folate‐deficient culture medium ,which might be related to PI3K/AKT/mTOR signal pathway .