Objective:To construct prokaryotic expression vector of mutant nm23-H1(S120G,S120G-P96S, S44A), and then express and purify the proteins. Methods: Mutant nm23-H1 fragments were amplified by PCR. The prokaryotic expression vectors of pET28a-nm23-H1 were constructed by gene recombination technique and verified by restriction enzyme analysis and sequencing. The positive clones were transformed into E. coli BL21(DE3) and soluble analysis of the expression was conducted in these systems. The proteins were purified by nickel column chromatography and identified by Western blot. Results: The sequences and open read frames of all the pET28a-nm23-H1 were completely correct. After transforming, these plasmids could express the target proteins. The protein production was very high. Among these mutant proteins, S44A was soluble expression and S120G was partly soluble. However, S120G-P96S was mostly in the inclusion bodies. The molecular weight of these mutant proteins was 20 kD detected by Western blot, which was as the same as the objective protein. Conclusion: We have succeeded in constructing the prokaryotic expression vectors of pET28a-nm23-H1(S120G, S120G-P96S, S44A), and the S120G and S44A proteins can be used in following studies. However, S120G-P96S protein which expressed in this system may not be suitable for the following studies.

As a common and frequently encountered dermatosis, psoriasis has been attracted the attention of dermatologists worldwide. In the past 30 years, radioimmunoassay, immunohistochemistry and molecular biological techniques were employed to determine the holistic levels of neuro immune protein (NIP), cyclic adenosine monophosphate (cAMP), cyclic guanosine monophosphate (cGMP), and cAMP/cGMP ratio in epidermis and dermis and body fluid (blood and urine), as well as the expression status of CD44, heat shock proteins (HSP) and nerve growth factor (NGF) in skin lesions of the patients with psoriasis and the healthy volunteers. The regulating function of autonomous nervous system was examined by analyzing the variability of heart rate (HRV). In addition, the sleeping status of psoriasis patients was compared with that of normal subjects by polysomnography. Furthermore, a series of surveys from the psychological prospect, including type A personality, depression, anxiety disorders and stressful life events, were performed among psoriasis patients. All the basic researches and clinical studies listed above have indicated that psoriasis is a kind of psychosomatic disease; and psoriatic patients are suffering from a maladjustment of the holistic regulating function in different degrees. It has been proved in the long-term clinical practices that the side effect of the total medicinal therapy exceeds the positive effect. It is recommended that great efforts should be given to provide the patients with effective cognitive education and psychological therapy. As a result, patients can release their tense and mental pressure by themselves. In summary, on the basis of a safe and simple medication therapy and well adjustment of holistic status, psoriasis patients can achieve satisfactory effect, prolonged duration of remission and improved their quality of life.

Thermotherapy is one of the combined therapies of malignant tumor. It can enhance the sen-sitivity to radiotherapy by inhibiting cell DNA repair,changing cell signaling, modulating cell cycle checkpoint and inhibiting tumor cell metastasis. Many clinical trials have also confirmed that the combination of thermother-apy and radiotherapy can enhance the curative effect of head and neck,ventrum and pelvic malignant tumor. In this article,the synergistic effects and mechanism of tumor thermoradiotherapy are reviewed.

Objective To explore the multi-resistance pattern of 88 strains of isoniazid-resistant Mycobacteria in recent years,so as to provide reference for prevention and treatment of tuberculosis.Methods Absolute concentration method was applied to performing drug susceptibility test for 181 isolates of Mycobacteria to 10 kinds of antitubercuIosis drugs. Results The primary resistance rate was 19.3%,and the acquired drug resistance rate Was 55.8%.The percentage of isoniazid-or double resistant-Mycobacteria was 37.6%.The Mycobacterium which was resistant to three or above drugs accounted for 85.2 %.Conclusion The majority of isoniazid-resistant Mycobacterium is a multi-resistant strain,which should be thought highly in prevention and treatment of tuberculosis to control or reduce the initial and acquired resistance rates.

[Objective] To explore the culture conditions of inducing callus from tissue of Aristolochia contorta Bge (ACB). [Methods] The leaves of ACB were used as the explants. Basic medium (including 1/2 MS medium, MS medium and modified MS medium) containing corresponding phyto-hormones was applied for the induction of ACB callus. The influerices of different culture conditions such as three kinds of basic medium, different pH values and addition of different kinds of phyto-hormones at different concentrations into the basic medium respectively or simultaneously, on the callus induction of ACB were observed. [Results] (1) After the three kinds of basic medium were added with the phytohormones of 0.1mg/L kinetin (KT) and 0.2mg/L ?-naphthalene acetic acid (NAA) , a large amount of ACB calluses were induced in the MS medium and modified MS medium, while ACB calluses did not occur in the 1/2 MS medium. (2) When the pH value of the culture medium composed of modified MS medium and 0.1mg/L KT and 0.2mg/L NAA was at 5.8, 7.0 and 8.0, a large amount of ACB calluses were induced in the medium with pH value being 7.0 and 8.0 while a few calluses occurred in the medium with pH value being 5.8. (3) ACB calluses were induced in modified MS medium with NAA added , but calluses did not occur in modified MS medium with KT added. When the modified MS medium was added with different kinds of phyto-hormones at different concentrations, ACB calluses were loose in the medium with high-concentration NAA and this did not benefit to the differentiation of buds for too fast cell division, and the callus induction rate was low in the medium with low-concentration NAA. The optimized culture medium was modified MS medium with 1mg/L KT and 0.5mg/L NAA added. [Conclusion] The optimum culture conditions of inducing callus from ACB tissues are: MS medium or modified MS medium with 1mg/L KT and 0.5 mg/L NAA added being the culture medium, and pH value being 7.0-8.0.

Objective To study the effect of azone,oleic acid,and their combination on the trans-dermal obsorption of baicalin.Methods Improved Franz-cells and HPLC were used for the transdermal study.The effect of azone,oleic acid,and their combination on the transdermal absorption of baicalin was studied using uniform design.Results The 24 h penetration amount of baicalin was obtained as 466.11 ?g/cm~2 and 77.34 ?g/cm~2 when the azone concentration was 10% or the oleic acid concentration was 14%,respectively.When using a combination of azone and oleic acid in the ratio of 10%∶0.5%,the 24 h penetration amount of baicalin was 641.88 ?g/cm~2 and the enhancement ratio of penetration was 146.69.Conclusion Both azone and oleic acid could improve the skin penetration amount of baicalin.Combinated application of azone and oleic acid could further enhance baicalin transdermal delivery rate.The maximum penetration amount of baicalin is obtained by using a combination of azone and oleic acid in the ratio of 10%∶0.5%.

Objective To investigate the expression patterns of artemisinin biosynthetic genes in Artemisia annua L.during the development stage and in different tissues,and to explore the mechanism of spatial and temporal modulators for artmisinin production.Methods The transcriptional profiles of artemisinin biosynthetic genes in the capital and cooperative pathways were quantitatively assayed by real time fluorescence quantitative-polymerase chain reaction(RTFQ-PCR) in different tissues of roots,stems,leaves and flowers,and in leaf-flourishing,pre-floral,and post-floral periods.Results The expression levels of the tested genes were extremely low in June,raised in July,and reached their peak values in August(before flowering),but dropped gradually in September(after blooming).In August,the transcription levels of the tested genes increased by 3 to 15 times compared to the lowestlevels.In particular,ADS and CYP71AV1 mRNA levels had the great elevation,which were 12 and 15 times as much as those in June.During the flowering period,the artemisinin biosynthetic genes mRNA expression was detectable in roots,stems,leaves and flowers,and the expression levels had no obvious difference except that ADS mRNA level in leaves was 2 times higher than that in other tissues(P

Two kinds of incorrect cultivation modes were existed since the launch of cultivation for professional degree postgraduate of clinical medicine:taking professional degree postgraduates as scientific degree postgraduates to cultivate or taking scientific degree postgraduates as professional degree postgraduates to cultivate.In this paper,taking the cultivation of oncologic professional degree postgraduates as example,we put forward some suggestions:strengthening clinical skill training,promoting thinking mode of evidence-based medicine and enhancing clinical trial and statistical training.

Objective To study the effect of the evidence-based nursing in the treatment of acute myocardial infarction by the early thrombolytic therapy with urokinase.Method 138 patients with acute myocardial infarction were randomly divided into the observation group and control group in equal number: the former received the evidence-based nursing and the latter conventional nursing care.The two groups were compared in terms of recanalization rate,fatality and incidence of angina.Result The recanalization rate,fatality and incidence of angina in the observation group were all significantly lower than those in the control group(all P<0.05).Conclusion The evidence-based nursing used to nurse the acute myocardial infarction patient to be treated by is cared by early thrombolytic therapy with urokinase may prevent such complications as angina and reduce recanalization rate and fatality.

Purpose:To investigate the effect of in vitro antisense oligodeoxynucleotide of cyclin D 1 on the cyclin D 1 gene expression and cell proliferation of human stomach adenocarcinoma cell BGC 823 cell line.Methods:Phosphorothioate cyclin D 1 ASODN and random oligodeoxynucleotide (RODN) were synthesized and transfected into BGC 823 Cells. Their effects on cell proliferation were examined by MTT method,RT PCR method,immunohistochemical study.Results:Cyclin D 1 ASODN could significantly inhibit the growth of BGC 823 Cell lines.The RODN showed no such effect.The inhibition peaked at 48 hour after transfection by MTT method and was dose dependent.ASODN could downregulate the expression levels of cyclin D 1 mRNA and protein by RT PCR method and immunohistochemical study respectively.Conclusions:The data suggested that ASODN could specifically inhibit the expression of cyclin D 1 mRNA and protein and regulate cell cycle and cell proliferation of BGC 823 cells. [