According to the sequence of P450 cDNA of Eleutherococcus senticosus, specific primers were designed. Frokaryotic ex pression vector pET30a-P450 was constructed and the prokaryotic expression conditions were optimized. Results showed that the BL21 after being transformed with the recombinant expression vector accumulated the high amount of recombinant protein. SDS-PAGE analysis showed that the recombinant protein was about 53 kDa. The recombinant accumulated the highest amount of recombinant protein af ter IPTG (1 mmol x L(-1)) at 27-37 degrees C for 24 h. Consequently P450 gene of E. senticosus could be expressed successfully by prokaryotic expression vector pET30a-P450. Induction temperature, IPTG concentration, medium type and amount of induction time could all influence the expression of target protein, but the impact strength was different.
Cloning, Molecular ; Cytochrome P-450 Enzyme System ; genetics ; metabolism ; Eleutherococcus ; enzymology ; genetics ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; Genetic Vectors ; genetics ; metabolism ; Plant Proteins ; genetics ; metabolism ; Plasmids ; genetics ; metabolism ; Recombinant Proteins ; genetics ; metabolism

In this study,we discussed the way of imparting the microbial knowledge and elevating non-biological student's science in public optional course "Microbes and Man" from the teaching practice.During the course,we first chose the material according to students' characteristic,and then introduced several methods like lecturing on subject,visiting factories and making microbial experiments in order to inter-est students in microbiology and get better in teaching effects.

Commercial production of bioethanol from lignocellulosic hydrolysates requires efficient fermenting strains. The abilities of the strain to converting all types of sugars in the hydrolysate to ethanol in high yield and to effectively tolerating/metabolizing inhibitors are necessary. Genome shuffling is a novel method for breeding, and it has been applied in pharmaceutical and food industry. This review summarized the technique of genome shuffling including principle, process, applications and its prospect for strains improvement in ethanol production from lignocellulosic hydrolysates.

There are abundant of lignocelluloses in agricultural wastes. And it can provide a variety of sugars involve pentose and hexose through pretreatments or hydrolyzation of these lignocelluloses. This review summarized the current status of bioethanol production by bacteria, and compared the productivity of some kinds of ethanologenic bacteria such as recombinant Zymomonas mobilis and Escherichia coli.

Objective: To observe the clinical efficacy of spleen-invigorating and qi-benefiting pediatric massage (tuina) for treating recurrent respiratory tract infection in children with cerebral palsy due to qi deficiency of spleen and lung. Methods: A total of 70 children with cerebral palsy who suffered from recurrent respiratory tract infection due to qi deficiency of spleen and lung were randomized into an observation group and a control group by the random number table method, with 35 cases in each group. Both groups were treated with conventional rehabilitation training, while the observation group was given additional spleen-invigorating and qi-benefiting pediatric massage, and the control group additionally took oral Yu Ping Feng granule. The clinical efficacy was evaluated by the total effective rate, traditional Chinese medicine (TCM) symptom score, and serum levels of immunoglobulin (Ig) A, IgM and IgG. Results: The difference in total effective rate between the two groups after treatment was statistically significant (P<0.05). After treatment, the TCM symptom scores in both groups decreased to varying degrees than those before treatment, and the intra-group differences were statistically significant (all P<0.01); the differences in the scores of various TCM symptoms between the two groups were statistically significant (all P<0.05). After treatment, the levels of IgA, IgM and IgG of the children in both groups increased to varying degrees, and the intra-group differences were statistically significant (all P<0.05). The between-group differences in the IgA, IgM and IgG levels after treatment were statistically significant (all P<0.05). Conclusion: Spleen-invigorating and qi-benefiting pediatric massage can effectively treat recurrent respiratory tract infection due to qi deficiency of spleen and lung in children with cerebral palsy, relieve the clinical symptoms and improve immune function, and thus is worthy of clinical promotion and application.

Opioid receptor, is classified into three subtypes, mu, kappa and delta, with the mu-type receptor plays important roles in opioid analgesia and opioid addiction. The cDNA encoding mu-type receptor was obtained by RT-PCR from human brain RNA and was cloned into pcDNA3.1(+). The resultant recombinant plasmid pcDNAMORs were transfected into CHO cells by liposome. After PCR identification, the positive clone were treated with agonist and antiagonist were tested for their competence of signal transduction. CHO cells that contained mu-opioid receptor in the expression vector pcDNA3.1(+) acquired naloxone-blockable high-affinity specific binding of morphine and DAMGO. The concentration of cAMP in CHO cells transfected with pcDNAMOR was reduced after binding to morphine and DAMGO, and increased after binding naloxone. These results indicate that the mu-type receptor expreesd on the CHO cell has similar biological property as the nature receptor. The availability of these specific cell lines will facilitate the drug development and promote our understanding the mechanism underlying opiate addiction.
Animals ; Brain Chemistry ; CHO Cells ; Cricetinae ; Cricetulus ; DNA, Complementary ; biosynthesis ; genetics ; Humans ; Receptors, Opioid, mu ; biosynthesis ; genetics ; Transfection

Adolescent ; Child ; Child, Preschool ; Chromosomes, Human, Pair 6 ; Female ; Humans ; Infant ; Loss of Heterozygosity ; Male ; Microsatellite Repeats ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics

B7-1 Antigen ; genetics ; immunology ; Carcinoma, Hepatocellular ; genetics ; immunology ; Enterotoxins ; genetics ; immunology ; Genetic Vectors ; Humans ; Liver Neoplasms ; genetics ; immunology ; T-Lymphocytes ; immunology ; Transfection ; Tumor Cells, Cultured

OBJECTIVETo explore the trend of influenza epidemics, predominate strains of the virus in Tianjin city and to analyze the economic impact of vaccine-based interventions for the prevention and control of influenza.

METHODSData on epidemiological studies and on virus surveillance was gathered. Monte Carlo mathematical simulation modelwas used for data analyses.

RESULTSFrom 29, December, 2003 to 2, January, 2005, the proportion of influenza-like infection cases was accounted for 8.93% of the total number of patients from the outpatient departments of 4 general hospitals in Tianjin. The proportion reached its peak from November to February and it was accounted for 9.39% in epidemic period, which was significantly different from that in the non-epidemic period (u = 15.53, P< 0.05). There was 56 strains of influenza virus isolated in which 45 were indentified as A(H3N2) and 11 as B with a total positive rate as 13.21%. Predominant strain was found bing type A(H3N2). The positive rate was 15.41% in the epidemic period, which was significantly different from that in the non-epidemic period (u = 2.519, P < 0.05). The cost per visit ranged from 475.93 to 581.69 Yuan (RMB) with an average cost of 528.81 Yuan. When the attack rate increased to 30 percent, the positive mean net returns would have been 24 million Yuan among the 0-19 age group.

CONCLUSIONInfluenza did not seem to be prevalent in Tianjin from 29, December, 2003 to 2, January, 2005. The main type of influenza was type A(H3N2). It is necessary to pay attention to the all-year round surveillance program due to the transformation of influenza type A and the slight increase of incidence in summer.

Adolescent ; Adult ; Child ; Child, Preschool ; China ; epidemiology ; Health Care Costs ; statistics & numerical data ; Humans ; Infant ; Infant, Newborn ; Influenza A Virus, H3N2 Subtype ; physiology ; Influenza B virus ; physiology ; Influenza, Human ; economics ; epidemiology ; immunology ; virology ; Middle Aged ; Population Surveillance ; Seasons ; Viral Vaccines ; immunology

Objective:To observe the effect of Governor Vessel-unblocking and mind-refreshing acupuncture plus functional training on neural development in infants with brain damage and seek an effective method for early intervention of infantile brain damage.Methods:Eighty infants with brain injury were recruited and allocated to a treatment group and a control group by their visiting sequence,with 40 cases in each group.The control group received exercise training,40 min each session and 6 sessions a week,and tuina treatment,30 min each time and 6 times a week.Based on the treatment protocol for the control group,the treatment group additionally received Governor Vessel-unblocking and mind-refreshing acupuncture,3 times a week and 10 sessions as a course at a 2-week interval.Before the treatment and after 14-week treatment,the gross motor function measure (GMFM) and developmental quotient (DQ) of Bejing Gesell developmental scale were used to evaluate the development of the infants.Results:After the treatment,the GMFM score and DQs of Gesell scale all increased by different levels in the two groups,and the intra-group differences were statistically significant (all P＜0.05);the scores of the treatment group were superior to those of the control group,and the between-group differences were statistically significant (all P＜0.05).Conclusion:Governor Vessel-unblocking and mind-refreshing acupuncture plus functional training can significantly promote the development of gross motor and cognitive functions in infants with brain damage,and it is an early and effective intervention for infantile brain damage.