OBJECTIVE:To prepare Luhuifangba burn cream and establish a method for its quality control.METHODS:The burn cream was prepared from freeze-dried powder of aloe barbadensis miller gel,borneol,hexadecanol,cosmolin,sodium dodecylsulfate,glycerol,ethylparaben and distilled water.Polysaccharide in the preparation was identified by paper chromatography,and borneol was identified by means of the color reaction between1%vanillin-sulfuric acid and bornyl alcohol and isoborneol.The content of polysaccharide in the cream was determined using anthrone-sulfuric acid method.RESULTS:The preparation was a cream in ivory white color,which spread easily and smelled slightly oily.Polysaccharide and borneol in aloe were able to be identified by the above-mentioned method.The linear range of polysaccharide was0.02～0.2mg/ml,and the average recovery was106.3%(RSD=2.5%).CONCLUSION:The preparation method is feasible,and the quality control method is simple,accurate,and reproducible.

Objective To observe the expressions of antimicrobial peptides LL-37,human beta defensin2 (HBD-2) and HBD-3 in the lesions of lupus vulgaris and tuberculosis verrucosa cutis,and to speculate the pathogenesis of cutaneous tuberculosis.Methods Tissue specimens were collected from the lesions of 18 patients with lupus vulgaris or tuberculosis verrucosa cutis and 15 patients with psoriasis,as well as from the normal skin of 10 healthy controls.The specimens were embedded by paraffin and subjected to an immunohistochemical analysis for the detection of antimicrobial peptides LL-37,HBD-2 and HBD-3.Statistical analysis was performed by using the SPSS 13.0 software package,and t test was carried out to compare the expressions of LL-37,HBD-2 and HBD-3 between these groups.Results LL-37 and HBD-2 were observed mainly in the upper and middle layer of the epidermis,appendages and vascular endothelial cells in cutaneous tuberculosis lesions.The expression levels of LL-37 and HBD-2 were significantly higher in tuberculosis lesions than in the normal skin (t =2.632,2.399,both P ＜ 0.05).Psoriatic and tuberculosis lesions shared a similar expression pattern for LL-37 and HBD-2.HBD-3 was absent in the lesions of cutaneous tuberculosis,but present in both psoriatic lesions and normal skin.Conclusions Both LL-37 and HBD-2 may participate in the immune response in cutaneous tuberculosis,while the absence of HBD-3 may contribute to the pathogenesis of cutaneous tuberculosis.

ObjectiveTo investigate the role of electroencephalogram (EEG) examination in the prognosis evaluation of patients with severe craniocerebral injury.MethodsFifty-seven patients with severe craniocerebral injury were selected as study subjects and their clinical data were analyzed retrospectively.All the patients got EEG examination,and the relationships between EEG grading and Glasgow coma scale (GCS) score,the prognosis of outcome were analyzed.ResultsThe EEG of 57 patients within 24 h after hospitalization were all abnormal.There was significantly negative correlation between EEG grading and GCS score (r =-0.742,P ＜0.05).EEG grading was significantly positively correlated with the prognosis of outcome (r =0.730,P＜ 0.05).ConclusionsEEG examination and EEG grading can evaluate the prognosis of patients with severe craniocerebral injury.It provides reference for clinical treatments.

Objective To observe the expression of Toll-like receptors (TLRs) 2,4 and 9 in lesions of lupus vulgaris and tuberculosis verrucosa cutis,and to evaluate the possible roles of these receptors in the pathogenesis of cutaneous tuberculosis.Methods Skin specimens were obtained from the lesions of 18 patients with clinically and pathologically diagnosed lupus vulgaris or tuberculosis verrucosa cutis,15 patients with plaque-type psoriasis (positive control),and from perilesional normal skin of 10 patients with pigmented nevi (negative control).Immunohistochemistry was used to detect the expression of TLRs 2,4 and 9 in these specimens,and the expression intensity was expressed as absorbence values.Data were analysed by using the SPSS 13.0 software,and t test was conducted to assess the differences between the three groups of specimens.Results TLR-2 was observed mainly in the middle and upper layer of epidermis in lesions of cutaneous tuberculosis and psoriasis,and in the whole epidermis except the basal layer in normal skin,with no significant difference in the expression intensity between the three groups of specimens (0.25 ± 0.04 vs.0.25 ± 0.05 vs.0.28 ± 0.03,P ＞ 0.05).TLR-4 was weakly expressed in cutaneous tuberculosis lesions,absent or slightly expressed in normal skin and psoriatic lesions,and the expression level of TLR-4 was significantly higher in cutaneous tuberculosis lesions than in normal skin and psoriatic lesions (0.16 ± 0.07 vs.0.07 ± 0.09 and 0.02± 0.05,t =2.58,6.24,P ＜ 0.01 and 0.05).TLR-9 was expressed in the whole epidermal layer,including appendages and vascular walls,of cutaneous tuberculosis and normal skin,with a significant increase in cutaneous tuberculosis than in the normal skin (0.25 ± 0.05 vs.0.19 ± 0.05,t =2.88,P＜ 0.05).Conclusions TLRs 2,4 and 9 are all expressed in cutaneous tuberculosis losious,and the expression intensity of TLR-4 and TLR-9 is higher in cutaneous tuberculosis lesions than in normal skin,hinting that TLR-4 and TLR-9 play a certain role in the immune response of cutaneous tuberculosis.

Intestinal flora is an important component of the body and is closely related to the health of infants and toddlers.There are many hypotheses about the origin and establishment mechanism of intestinal flora, but most theories are still controversial.The intestinal flora is not static, and its biodiversity and abundance will continue to differentiate and evolve with the growth and development of nurslings.It is now believed that intestinal flora plays a variety of roles in maintaining the homeostasis of the infant's internal environment.This review summarizes the basic research on the physiology of infantile intestinal flora in recent years, to provide some reference for the development of this branch in the field of pediatrics.

OBJECTIVE： To establish a UHPLC method for determination of the contents of 11-keto-β-boswellic acid(KAB) and 11-keto-β-acetyl-boswellic acid(AKBA)in Frankincense and explore the suitability and accuracy of substitute reference substance method with DRS origin software for qualitative and quantitative determination of chromatographic peaks. METHODS ：The samples were separated by UHPLC for determination of AKBA and KBA. AKBA was used as a reference to investigate the accuracy of KBA identification using DRS origin software on 19 different C18 columns. The RSDs of relative correction factors were calculated for different detection wavelengths and instruments.The relative correction factor method and the external standard method were selected for quantification and the differences were compared. RESULTS: The established method met the requirements of methodology and the average recovery was 100.21%(n=6) with RSD of 2.47%. The DRS origin software can be used to accurately determine the chromatographic peaks. The correct factor of AKBA vs. KBA was 0.936 and it was consistent under different conditions. There were no significant differences between the content calculated by the relative correction factor method and by the external standard method. CONCLUSION： This method is intelligent, feasible, reliable and economical, and can be used for the determination of frankincense content.

The genes of short armed hammerhead ribozyme targeting against two sites on positive strand (194-196) and negative strand (89-91) of ASSVd were designed, synthesized and cloned according to the action manner of hammerhead ribozyme. The full lengths of the genes are 42 bp (RzASSVd(+)) and 40 bp (RzASSVd(-)). After transcription in vitro, the ASSVd positive and negative RNA labeled with 32P were mixed with the ribozyme transcript and incubated 3-4 h at 50 degrees C or 37 degrees C. The results were assayed on 8% PAGE (containing 8 mol/L urea) and autoradiogrammed. As predicted, the transcript of the active RzASSVd(-) could cleave the ASSVd negative strand RNA with a high activity but had no cleavage effect on the ASSVd positive strand. The transcript of the RzASSVd(+) gene could cleave the ASSVd positive strand but its cleavage activity was very low. As the same, it cannot cleave the negative strand either. On the base of the result, we construct dimmer ribozyme gene pGEMRzASSVd(+/-) containing both RzASSVd(+) and RzASSVd(-).
Cloning, Molecular ; Malus ; virology ; Plant Diseases ; virology ; RNA, Catalytic ; genetics ; metabolism ; therapeutic use ; RNA, Viral ; metabolism ; Substrate Specificity ; Transcription, Genetic ; Viroids ; metabolism

BACKGROUND:Exogenous basic fibroblast growth factor (bFGF) plays an important role in the ligament tissue healing process, and the use of transgenic methods to transfect exogenous genes into cells can promote the secretion of bFGF. OBJECTIVE:To observe phenotypic changes and the bFGF protein expression after bFGF recombinant adenovirus was used to transfect rabbit bone marrow mesenchymal stem cells (BMSCs). METHODS:Passage 2 BMSCs were divided into three groups:Ad.bFGF-eGFP group, Ad.eGFP group and control group. Under a phase contrast microscope we observed the changes in cellmorphology. The expression of bFGF protein in BMSCs was determined by enzyme-linked immunosorbent assay (ELISA). The proliferative curve was detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). RESULTS AND CONCLUSION:The transfected cells showed a uniform phenotype of fibroblasts. MTT colorimetric assay revealed that more proliferative activity of transfected BMSCs was shown in the Ad.bFGF-eGFP group than in the Ad.eGFP group and control group. ELISA results showed that expression of bFGF protein was higher in the Ad.bFGF-eGFP group than in the Ad.eGFP group and control group (P<0.05). BFGF recombinant adenovirus can induce the differentiation of BMSCs into fibroblasts, increase proliferative ability and promote the expression of bFGF protein.

Objective To investigate the typical value of gamma-radiation dose rate and its confidence interval in tank-transported copper ore by using bootstrap resampling techniques. Methods Bootstrap resampling method, coupled with kernel density estimation, introduced to acquire the typical value of gamma-radiation dose rate in copper ore. Results The typical value of gamma-radiation dose rate in copper ore was expressed as the central tendency of the means of resampling, and two kinds of confidence interval, empirical percentile and bias-corrected accelerated confidence interval, were provided as standard error. Conclusion It is clearly demonstrated that this method has an advantage to give a robust description in explanation of central tendency and variation range of gamma-radiation dose rate data profiles.

Objective To investigate the dynamical changes of Rho kinase in the cerebrospinal fluid (CSF) and its relationship with cerebral vascular spasm CVS. Methods CSF were collected on the ist, 3rd, 7th, 10th and 14th day after subarachnoid hemorrhage. The expression of Rho-kinase mRNA in CSF was determined by RT-PCR. The expression of endothelin-1 in CSF was determined by radioimmuno-assay. TCD was used to measure the velocity of the cerebral artery. Results The levels of ET-1 and Rho-kinase mRNA in CSF were re-markably increased on the 3rd day, and reached at the peak on the 7th day after subarachnoid hemorrhage, which were significantly higher than those without CVS. Conclusion There is a positive correlation between the level of Rho-kinase mRNA and ET-1 in CSF. Rho-kinase may participate in the development of CVS.