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.

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 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.

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.

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

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.

Objective To explore the correlation between CT perfusion imaging parameters and differentiation degree of gastric cancer.Methods Fifty patients with gastric cancer proved by surgery and pathology underwent 64-slice spiral CT perfusion imaging before surgery,and the CT perfusion parameters were obtained including blood flow(BF),blood volume(BV),mean transit time (MTT)and permeability surface(PS).The patients were divided into 3 groups (well,moderate and poor differentiation)according to the degree of cell differentiation.Statistical analysis was performed by using the SPSS 17.0 statistics software.Results The perfusion parameters of BF (mL·min-1 ·100 g-1 ),BV (mL/100 g),MTT (s)and PS (mL·min-1 ·100 g-1 )were as follows:75.28±6.81,9.01± 0.94,9.89±1.65 and 10.05±0.71 in well differentiation group with 10 patients (20%),110.01±31.90,18.18±5.62,9.81±3.69 and 40.08±1 5.82 in moderate differentiation group with 24 patients (48%),138.59±38.09,21.08±4.1 1,9.47 ±1.80 and 57.50± 13.28 in poor differentiation group with 1 6 patients (32%)respectively.Among three groups,there were statistic differences in BF, BV and PS between well differentiation group and moderate differentiation group or low differentiation group (P <0.05),however there was no statistic difference in MTT among three groups (P >0.05).Conclusion The BF,BV and PS values may be regarded as the evaluation index for malignant degree of gastric cancer.

BACKGROUND: The association of tumor necrosis factor alpha (TNF-α) gene polymorphisms with the onset and development of ankylosing spondylitis (AS) has been the focus of studies on AS in the field of genetics.OBJECTIVE: To explore the association of the polymophisms of TNF-α promoter gene at positions-308 and -238 with AS susceptibility and clinical pathological changes.DESIGN: A case-control study.SETTING:The Rheumatic Immunology Department of Changhai Hospital of the Second Military Medical University of Chinese PLA.PARTICIPANTS: Totally, 108 AS patients were recruited from Rheumatic Immunology Department of Changhai Hospital, Second Military Medical University of Chinese PLA from January 1999 to December 2003 ,they had no kinship. The ratio of men to women was 5.3: 1. They aged from 13 to 71 (30-± 12) years old, and AS was divided into Ⅰ- Ⅳ radiographic stages according to the sacro-iliac joint damage. A total of 100 healthy controls were randomly selected from the blood donators(Shanghai Hospital) who were aged from 19 -56 (33 ±9) years old, and the ratio of men to women was 4.9: 1. Informed consent was obtained from all the subjects.ti-coagulated with EDTA. Polymerase chain reaction amplification and purification of the TNF-α promoter region was made and the sequence of polymerase chain reaction products was examined and displayed by Chromas 1.62 softcorresponding radiographic stage of sacro-iliac joint damage was assessed to investigate the influence of gene polymorphisms on AS.MAIN OUTCOME MEASURES: DNA direct sequencing method was used to detect -238 and -308 allele phenotypes for investigating the association with clinical presentations.G and -238G/A allele was 98.1% (106 cases) and1. 9% (2 cases) respectively in AS group and 95.0% (95 cases) and 5.0% (5 cases) respecquency of TNF-α promoter gene at positions -308. 1.1(G/G) and - 308.1.2(G/A) alleles was 82.4% (89 cases) and 17.6% (192 cases) respectively in AS group, which was not significantly different compared respectively with 85.0% (85 cases) and 14.0% (14 cases) of the control of sacro-iliac joint damage and the frequency of TNF-α promoter gene at the position of - 308 (G/G) and (G/A): AS patients with(G/G) phenotype who were confirmed of radiographic stage Ⅰ, Ⅱ, Ⅲ, and Ⅳ were observed in 3/35/40/11cases,compared with (G/A) phenotype of 1/12/6/0 cases.The difference was statistically significant (χ2GMH = 4.77, P ＜ 0.05 ).CONCLUSION: Our data suggest that the polymorphisms of TNF-α promoter gene at positions of - 238 and - 308 allele has no association with AS susceptibility, but the polymorphisms of TNF - α promoter gene at the position of -308 might exert great influence on AS according to the radiographic stage of sacro-iliac joint damage.