A highly sensitive immuno-PCR assay based on sandwich ELISA and PCR was developed to detect the circulating antigen in trichinellosis. Antigens were purified from the muscle larvae of Trichinella spiralis, and the myeloma cells were fused with spenocytes immunized with T. spiralis antigens to product the specific monoclonal antibodies. Indirect ELISA was used to select the antibody-secreting hybrodoma cells. By this method of procedure, monoclonal antibody F4C6 against the T. spiralis ES antigen was obtained, which was used as the indicator antibody, while the rabbit polyclonal antibodies against T. spiralis were to be used as capturing antibodies. The plasmid Bluecript Ⅱ KS was amplified by PCR with biotin-labeled primer M13-20, and thus the biotin-labeled DNA was obtained. Both the second antibody and DNA labeled with biotin were to be linked with 100 ng/ml avidin. The whole procedures of assay consisted of two steps, in which the circulating antigens were captured by monoclonal antibody through sandwich ELISA in the first step, and the DNA linked by monoclonal antibody was amplified by PCR in the second step. The sensitivity of this method was compared with that of the ELISA assay. It was found that the measuring ranges to detect the circulating antigens in trichinellosis were 50 pg/L to 0.005 pg/L for the immuno-PCR assay, and 5 μg/L to 0.05 μg/L for ELISA assay, the former was quite higher than that of the latter. It is evident that this method is highly sensitive for the detection of circulating antigens in trichinellosis.

Child ; Child, Preschool ; Female ; Fluoroacetates ; poisoning ; Hemoperfusion ; Humans ; Infant ; Male

OBJECTIVEThe clinical results of restoring defective teeth with two types of glass fiber prothetic systems is investigated to acquire clinical experience for further application of glass fiber posts with independent intellectual property rights.

METHODSA total of 120 out-patients with restored defective teeth were selected from the Department of Stomatology, Beijing Xuanwu Hospital of Traditional Chinese Medicine and Peking University School and Hospital of Stomatology and randomly divided into two groups. OUYA FIBER posts and Tenax Fiber White posts were used to restore defective teeth in the experimental group and the control group, respectively. Clinical evaluation was conducted one week and three months post-restoration.

RESULTSBoth clinical satisfactory rates of OUYA FIBER posts and Tenax Fiber White posts were higher than 98% one week post-restoration and higher than 96% three months post-restoration. No significant differences were observed between the clinical results of restoring defective teeth with the two types of fiber posts. Throughout the healing process, no root canal fracture or marginal staining were observed.

CONCLUSIONOUYA FIBER post and Tenax Fiber White post showed similar clinical outcomes during the period of observation in this study. However, the long-term effects should be observed in future studies.

OBJECTIVE:To study the optimum formation technics of Xuesaitong drop pill.METHODS:Parallel tests were conducted on the dosage of different base materials and the main drug with the forming percentage and the rate of qualified weight as the index of evaluation,the orthogonal test was conducted on the4factors,including the temperature of drops and the liquor condensate,the drug height in the drug storage tank and the dropping distance.RESULTS:The ratio of base materials and the main drug was2.5∶1.The optimum forming technics could be seen as follows,the height of the drug storage tank was3cm,the temperature of drops was90℃,the dropping distance was5cm and the temperature of the liquor condensate was12.5℃.CONCLUSION:There was a high rate of end product of dropping pill prepared with this optimum process,which was in conformity with the standard stated in the Chinese Pharmacopoeia.

Objective To investigate the effects of photochemotherapy on the angiogenic ability of endothelial cells and expression of integrin. Methods In vitro angiogenesis assay ( UVA exposure dose: 0, 2.0, 5.0 J/cm~2) was used to detect the effects of photochemotherapy on the angiogenic ability of endothelial cells. Reverse polymerase chain reaction and flow cytometry were employed to determine the effects of photochemotherapy on the expression of integrin mRNA and protein, respectively. bFGF stimulation group and PMA stimulation group were divided according to the inductors, and subgroups were divided according to the UVA exposure doses of 0, 2. 0 and 5. 0 J/cm~2 in each group. Results Combination of UVA (2.0 and 5.0 J/cm~2) and 8-MOP (100 ng/mL) resulted in a decrease in the angiogenic ability of endothelial cells in vitro and the expression of integrin mRNA and protein. Conclusion Photochemotherapy may inhibit the angiogenic ability of endothelial cells through downregulating the expression of integrin.

Objective To investigate the relationship between the mutation of ERG11 gene,a target of azole antifungal drugs (fluconazole,itraconazole,voriconazole),and azole-resistance in Candida albicans isolates from patients with AIDS.Methods Ninety-three Candida albicans strains were isolated from patients with AIDS.DNA was extracted from these isolates,and ERG11 gene was amplified by PCR followed by bidirectional sequencing.DNAman software was used to compare the resultant sequence with the reference sequence of ERG11 gene (GenBank accession no.X13296).Then,different base sequences were translated into amino acid sequences to determine whether missense mutations occured.Results A total of 40 mutation sites were identified in these isolates,including 27 silent mutations and 13 missense mutations.One or no missense mutation was detected in Candida albicans strains resistant to 1 antifungal agent,while those resistant to 2 or 3 antifungal agents simultaneously harbored 2 or 3 missense mutations.Conclusion The missense mutations in ERG11 gene are probably connected with azole resistance in Candida albicans.

Objective To investigate the effect of all-trans-retinoid(ATRA) on the migration of endothelial cells and the expression of gelatinases. Methods In vitro migration assay was used to determine the effect of ATRA on the endothelial cell migration induced by phorbol 12-myristate-13-acetate (PMA), semi-quantitative RT-PCR and Western blot were used to observe the effect of ATRA on the gelatinase expression at mRNA and protein levels respectively, while the proteolytic activities of gelatinases were assessed by zymography. Results The endothelial cell migration elicited by PMA was significantly inhibited when incubated with 0.1 ?mol/L, 1.0 ?mol/L and 10.0 ?mol/L ATRA. Compared with the control, the inhibition rates were (44.68 ? 7.79)%, (65.20 ? 4.59)% and (78.37 ? 2.58)%, respectively. ATRA also reduced the expression and activities of gelationases in a dose dependant manner. At 10 ?mol/L concentration, the inhibition rate of mRNA expression, protein expression and protein activities for gelatinase A was (59.39 ? 7.98)%, (78.40 ? 3.23)% and (53.02 ? 7.23)%, respectively. For gelatinase B it was (65.23 ? 3.62)%, (82.49 ? 2.88)% and (47.32 ? 7.72)%, respectively. Conclusions The expression and activities of gelatinases are downregulated in the endothelial cells when incubated with ATRA, which may be the possible mechanism of ATRA inhibiting the endothelial cell migration elicited by PMA in vitro.

Objective To investigate the vole of comfort nursing in skin prick test. Methods 300 allergic rhinitis patients who received skin prick test from Jan. to Nov. 2009 in our department as control group were given regular nursing, while 303 allergic rhinitis patients from Jan. to Nov. 2010 as experimental group were received comfortable nursing care and care effects of both groups were compared. Results Compared with the control group, the occurrence rate of pain in the experimental group was significantly decreased, and there was statistical difference between both groups (X2 = 118, P ＜ 0. 01). Conclusions Comfort nursing care employed in skin prick test could significantly improve the quality of nursing care, reduce the pain of patients.

In order to evaluate the characteristics of the spray drying of total flavonoids of Epimedium extracts assisted with soybean polysaccharide, a certain percentage of soybean polysaccharide or polyvidone were added to the total flavonoids of Epimedium extract to conduct the spray drying. The effect of soybean polysaccharides against the wall sticking effect of the spray drying was detected, as well as the powder property of total flavonoids of Epimedium spray drying powder and the dissolution in vitro behavior of the effective component. Compared with the total flavonoids of Epimedium spray drying powder, soybean polysaccharide revealed a significant anti-wall sticking effect. The spray drying power which had no notable change in the grain size made a increase in the fluidity, improvement in the moisture absorption and remarkable rise in the dissolution in vitro behavior. It was worth further studying the application of soybean polysaccharide in spray drying power of traditional Chinese medicine.
Epimedium ; chemistry ; Flavonoids ; analysis ; Particle Size ; Polysaccharides ; chemistry ; Powders ; Soybeans ; chemistry

Objective To observe that antiretroviral efficacy, immune reconstitution of four-year highly active antiretroviral therapy (HAART), and evaluate its side effect in Chinese HIV-1-infected patients. Methods A total of 258 HIV-1 infected patients, given HAART regimens composed of two nucleoside reverse transcriptase inhibitor (NRTI) and one non-nucleoside reverse transcriptase inhibitor (NNRTI) for mean 51.5 months, measured HIV RNA viral load(VL) and the counts of CD4+ T cell,CD8+ T cell at the baseline and 6, 12, 24, 36 and 48 months after HAART initiation, respectively,monitoring side effect, blood routine, main biochemical parameters, and other disadvantageous accidents during the 51.5-month treatment. Results Plasma HIV-1 RNA level was determined by fluorescent quantitative polymerase chain reactions (FQ-PCR) at the baseline and 6, 12, 24, 36 and 48 months after starting HAART, and showed 5.27, 2.97, 2. 74, 2. 62, 2. 67 and 2.75 lg (copies/ml), respectively. The counts of CD4+ T cell from (127±63) cells/μl at the baseline increased to (190±115), (248±93),(269±127), (296 ± 156) and (317 ± 195) cells/μl at 6, 12, 24, 36 and 48 months after starting HAART. A total of 149 treated patients (57.8%)had gastrointestinal side effects, peripheral polyneuropathy, various rashes, central nervous system disorders, fever or baldness. Twenty-two patients changed one of three medicines to another because toxicity. Sixteen changed the regimen to the second line HAART for lactic acidosis or other serious toxicities. Conclusions A total of 258 HIV-1 infected Chinese patients treated with two NRTI and one NNRTI as first line HAART regimen during mean 51.5 months,showed a good antiretroviral efficacy and immune reconstitution, but a few site-effects at the parts of patients. It is necessary to treat adverse effect and change HAART regimen for severe toxicity in time.