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.

Objective]To observe the effect of abdominal acupuncture combined with body acupuncture on chloasma of stagnation of the liver-type. [Method] 30 patients were randomized into treatment group and control group, 15 in each. Treatment group were given abdominal needle combining acupuncture treatment, control group were only given acupuncture treatment. Each group were treated once a day, 5 times made a course, after each course had a rest for 2 days. After 4 courses, to observe the effect and the skin color score, and area score and declined index. [Results]After treatment, the two groups were effective, but the treatment group was more obvious, and scores difference compared with the control group was statistically significant(P<0.01). The total effective rate was 93.3% in the treatment group 86.67% in the control group, there was a significant difference between the two groups(P<0.05). [Conclusion]Abdominal hepatic stagnation type treated by acupuncture combined with body acupuncture on chloasma treatment, no pain or abdominal pain to a lesser degree, even more easily accepted by patients, is better than body acupuncture.

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 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 .

BACKGROUND:It is confirmed that the onset of osteoarthritis is closely related to inflammatory cytokines. OBJECTIVE: To investigate the levels of interleukin-6 and matrix metaloproteinase-3 in the synovial fluid of patients with osteoarthritis and their correlation with the severity of osteoarthritis. METHODS: Synovial fluid specimens of 78 patients were harvested to detect the levels of interleukin-6 and matrix metaloproteinase-3 using ELISA assay. The severity of osteoarthritis was estimated by the Hospital for Special Surgery (HSS) score. Pearson correlation analysis was adopted to explore the pairwise correlations between interleukin-6, matrix metaloproteinase-3 and HSS score. RESULTS AND CONCLUSION:The levels of interleukin-6 and matrix metaloproteinase-3 in the synovial fluid were (13.1±5.7) and (989.3±429.5) μg/L, respectively; the HSS score was (56.4±12.0) points. Pearson correlation analysis showed that the expression levels of interleukin-6 and matrix metaloproteinase-3 had a significant positive correlation (r=0.70,P< 0.001); the levels of interleukin-6 and matrix metaloproteinase-3 had a negative correlation with the HSS scores (r=-0.70,r=-0.75,P < 0.001). Experimental findings show that there is a positive correlation between interleukin-6 and matrix metaloproteinase-3 in the synovial fluids, and these two cytokines also have a positive correlation with the severity of osteoarthritis, indicating that to measure interleukin-6 and matrix metaloproteinase-3 levels in the synovial fluids of osteoarthritis patients is important for early diagnosis and treatment of osteoarthritis.

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.

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

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 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 investigate the distribution of intramuscular nerves in the different skeletal muscles of the rabbit. Methods Muscle architecture and modified sihler's neural staining methods were used. Results The nerve branches of innervating flat M.pectoralis major arose from the anterior pectoral and posterior pectoral nerve.The former innervated cross fibers mainly,it penetrated the middle of muscle belly and formed a “U-Shaped” nerve ansa.The latter innervated oblique fibers mainly.There were many anastomoses between them.The nerve of pennate M.plantaris derived from tibial nerve.After entering muscle,the nerve trunk gradually divided into many primary branches toward medial and lateral fibers,these branches then subdivided into numerous arboroid second and terminal branches toward all of muscle fibres;The nerve of innervating spindle M.extensor digitorum longus came from N.fibularis communis.Two extramuscular nerve trunk were seen.Superior trunk mainly innervated those muscle fibers of inserting in the second toe,Inferior one mainly distributed to the fibers of rest within this muscle.Conclusion The spatial arrangement of the muscle fibres were related to distribution of the intramuscular nerves;Walking of intramuscular nerve branches had two patterns which run perpendicular and/or parallel to muscle fasciculi.