Objective:To prepare a colloid gold kit for the qualitative detection of the dopes met-amphetamine and morphine by one-step chromatography immunoassay in human samples based on the principle of the highly specific immunochemical reactions between the antigens and their antibodies which were used for the analysis of specific compounds in human urine samples at ideal cut-off concentrations.Methods:The monoclonal antibodies(McAb) of either mouse anti-morphine or mouse met-amphetamine were prepared by hybridomas and the limiting dilution respectively. The titers of McAb were tested by ELISA.The kit was assembled and the specificity,sensitivity,and stability were investigated.In addition,these characters were compared with those of issued Pan Probe single test strip.Results:①The titer of the first McAb against morphine was 1∶3.2?103, as for another, 1∶1.6?103. ②The colloid gold kit was found with following cut-off for the dope concentrations,morphine at 300 ng/ml and met-amphetamine at 1 000 ng/ml. ③The colloid gold kit showed fine specificity without cross-reaction with routine medicines for cold treatment or with the solf-drinks of cokocola or with tobacco for the individuals who did not take the dopes the 7 days.④The colloid gold kit was even more sensitive than the issued strip,because urine sample of the junkies were still positive 4-5 days post-intake.Conclusion:The colloid gold kit for indentifying the drug-users is a rapid,specific, sensitive immunoassay suitable for the simultaneous, qualitative detection of drug abuse.

BACKGROUND: Umbilical cord blood mesenchymal stem cells (UCB-MSCs) can differentiate into various types of cells under certain condisions, and easily proliferate in vitro. However, UCB-MSCs have long establishment time and low frequency.OBJECTIVE: To in vitro isolate and culture UCB-MSCs, and induce its differentiation into osteoblasts.DESIGN, TIM E AND SETTING: The in vitro cytological study was performed at the Laboratory of the Medical College of Qingdao University from June 2008 to January 2009.MATERIALS: UCB was obtained from term normal delivery women at the Department of Gynaecology and Obstetrics, Qingdao Municipal Hospital.METHODS: Human UCB-MSCs were isolated and cultured in vitro by Percoll density gradient. When reached 90% confluency,UCB-MSCs were digested by trypsin for subculture. At the third passage, UCB-MSCs at 1×106 were incubated. When reached 50% 60% cenfluency, UCB-MSCs were treated with DMEM supplemented with 0.1 μmol/L dexamathasone, 10 mmol/Lβ-sodium glycerophosphate and 50 μmol/L vitamin C. UCB-MSCs in the control group were incubated in low glucose DMEM.MAIN OUTCOME MEASURES: Growth and proliferation of MSCs were observed under the inverted microscope. Cell surface marker expression and cell growth curve were measured by flow cytometry. Cell ultrastructure was observed under the transmission electron microscope. Differentiation of UCB-MSCs into osteoblasts was determined by Won Kossa staining and alkaline phosphatase staining.RESULTS: Primary cultured UCB-MSCs had similar morphology to bone marrow mesenchymal stem cells. After passage, cell morphology was even, presenting spindle shape. UCB-MSCs at passage 3 highly expressed CD29, CD44, CD13, but did not express CD34. Growth latency was 2-3 days. Cells entered logarithm proliferation phase at days 3-4, and platform phase 1 month later. Nuclei presented round or irregular, with clear nuclear membrane, 1-2 nucleoli, rough chromatin, abundant organelles and microvilli. UCB-MSCs at passage 3 were gradually confluent following 3 days of osteogenic induction, with the presence of pavement-stone shape. 14 days later, calcified nodules by Von Kossa staining, and cells were positive for alkaline phosphatase staining. In the control group, no calcified nodules were found, and cells were negative for alkaline phosphatase staining.CONCLUSION: UCB-MSCs can be successfully isolated by Percoil density gradient, and induced to differentiate into osteoblasts in vitro.

Oncomelania hupensis is the only intermediate host of Schistosoma japonicum. The elimination of Oncomelania snails is the key technique step for schistosomiasis control. This paper summarizes the progress of the techniques of snail control，including the methods of ecology engineering，biology，molluscicides and the study on novel molluscicides，and reviews their features. In addition，this paper explores the appropriate approach to control the snails.

Clinical practice is an important period to train student's skills in pediatric surgical learning,but it is confronted with difficult condition under the new doctor-patient relationship.How to resolve these difficulties is the key to carry on teaching successfully.Aaccording to the students' interests and the knowledge structure characteristics,teachers should cultivate their professionalism,train their communication skills,extend their thinking ability with appropriate use of teaching aid to increase the teaching interests and develop self-learning ability.

Education reform and innovative education are the strong requirements of the times and social development.We developed immunological paper free examination system based on the optimized question bank,characteristics of immunological examination and information technology.This system can randomly develop electronic examination paper and can integrate the processes of exam registration,examination paper development,online examination,paper inspection,scores generation and printing into one system,which can save human resources,enhance the accuracy and fairness of the examination,making it conform to international standards.

This paper discusses the problems of optimizing teaching of medical microbiol-ogy on the aspects of optimizing teaching matters and teaching means,of integrating teaching with learning,and training teachers,etc. to improve the teaching quality in microbiology through some innovational measures.

Objective To analyze the clinical features of severe pneumonia complicated with congenital heart disease (CHD) . Methods The clinical data of 270 children patients with severe pneumonia complicated with CHD (CHD group) were collected to analyze the relative medical history ,etiology ,bacterial drug resistance and clinical outcomes .Moreover ,636 age-matched children pa-tients with pure severe pneumonia were selected as the control group .Results In the CHD group ,250 cases were infants and 35 ca-ses suffered from repeated pneumonia .Compared with the control group ,the CHD group was earlier in onset and more prone to re-peated pneumonia .Among 270 cases ,totally 220 cases were checked out the pathogens ,but no statistically significant difference in the detection rates of pathogens between the CHD group and the control group was found (P>0 .05);179 cases were infected by bacteria ,in which 126 cases were Gram-negative bacteria ,the positive rate of bacterial infection and the detection rate of Gram-nega-tive bacterial were relatively higher than those in the control group ,the difference was statistically significant (P<0 .05);95 cases were infected by virus ,with respiratory syncytial virus being the most common pathogen ,30 cases were infected by fungi ,with Can-dida albicans being predominant ,the infection rates of virus and fungus had no statistical difference between two groups (P>0 .05);in the drug sensitive test ,the detection rate of extended spectrum beta lactamase (ESBLs)-producing positive bacteria in the CHD group was higher than that in the control group ,the difference was statistically significant (P<0 .05) .Main Gram-negative bacteria Pneumonia klebsiella pneumonia subspecies ,Escherichia coli and haemophilus inf luenzae were highly sensitive to imipenem ,mero-penem ,levofloxacin and ciprofloxacin ;main Gram-positive bacteria Staphylococcus aureus and streptococcus pneumoniae were highly sensitive to vancomycin and gentamicin .Especially ,pneumonia klebsiella pneumonia subspecies ,Escherichia coli and haemophilus influenzaewere100% resistanttopenicillin.TheaveragelengthofhospitalstayintheCHDgroupwas (20.9±12.5)d,207cases developed the respiratory failure ,88 cases appeared the heart failure and 205 cases were effective in treatment ;compared with the control group ,the CHD group had a longer length of hospital stay ,higher occurrence rate of heart and respiratory failure and lower treatment effective rate (P<0 .05) .Conclusion Because of the earlier onset ,more prone to repeated pneumonia ,more sensitive to bacterial infection ,esp .Gram-negative bacteria ,more ESBLs production ,longer hospital stay and more prone to complications ,com-pared with the control group ,children with severe pneumonia complicated with CHD require active treatment .

Some unhealthy life habits, such as long-term smoking, heavy drinking, sexual overstrain and frequent stay-up could induce the Yin deficiency symptoms of zygomatic red and dysphoria. Stems of Dendrobii officinalis flos (DOF) showed the efficacy of nourishing Yin. In this study, the hyperthyroidism Yin deficiency model was set up to study the yin nourishing effect and action mechanism of DOF, in order to provide the pharmacological basis for developing DOF resources and decreasing resource wastes. ICR mice were divided into five groups: the normal control group, the model control group, the positive control group and DOF extract groups (6.4 g · kg(-1)). Except for the normal group, the other groups were administrated with thyroxine for 30 d to set up the hyperthyroidism yin deficiency model. At the same time, the other groups were administrated with the corresponding drugs for 30 d. After administration for 4 weeks, the signs (facial temperature, pain domain, heart rate and autonomic activity) in mice were measured, and the facial and ear micro-circulation blood flow were detected by laser Doppler technology. After the last administration, all mice were fasted for 12 hours, blood were collected from their orbits, and serum were separated to detect AST, ALT, TG and TP by the automatic biochemistry analyzer and test T3, T4 and TSH levels by ELISA. (1) Compared with the normal control group, the model control group showed significant increases in facial and ear micro-circulation blood flow, facial temperature and heart rate (P < 0.05, P < 0.01), serum AST, ALT (P < 0.01), T3 level (P < 0.05), TSH level (P < 0.05) and notable deceases in pain domain (P < 0.01), TG level (P < 0.01). (2) Compared with the model control group, extracts from DOF (6 g · kg(-1)) could notably reduce facial and ear micro-circulation blood flow, facial temperature and heart rate (P < 0.05, P < 0.01) and AST (P < 0.05) and enhance pain domain (P < 0.01) and TG (P < 0.01). Extracts from DOF (4 g · kg(-1)) could remarkably reduce AST and ALT levels (P < 0.01, 0.05). Extracts from DOF (6 g · kg(-1) 4 g · kg(-1)) could significantly reduce T3 and increase serum TSH level (P < 0.05). DOF could improve Yin deficiency symptoms of zygomatic red and dysphoria in mice as well as liver function injury caused by overactive thyroid axis. According to its action mechanism, DOF may show yin nourishing and hepatic protective effects by impacting thyroxin substance metabolism, improving micro-circulation and reducing heart rate.
Animals ; Dendrobium ; chemistry ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Flowers ; chemistry ; Humans ; Hyperthyroidism ; drug therapy ; metabolism ; Male ; Mice ; Mice, Inbred ICR ; Phytotherapy ; Thyroxine ; metabolism ; Yin Deficiency ; drug therapy ; metabolism

AIM:To explore the molecular mechanism of panaxadiol saponin(PDS)by observing Toll like receptor(TLR)2 and TLR9 mRNA expression induced by lipopolysaccharide(LPS).METHODS:Rats were divided into LPS,LPS+PDSL,LPS+PDSM and control group,respectively.Nitric oxide synthase(NOS)activity,nitric oxide(NO)content,LPO content,SOD activity and TLR2 and TLR9 mRNA expression were assayed 4 h after intravenous injection of LPS.RESULTS:NOS activity,NO content,LPO content of LPS+PDSL group and LPS+PDSM group were significantly lower than those in LPS group.TLR2 mRNA expression in the liver tissue of LPS+PDSL group and LPS+PDSM group was decreased compared with LPS group.CONCLUSION:PDS has a protective effect on liver tissues by triggering the down-regulation of TLR2 expression,reducing NOS activity,and NO content.

Objective: To explore the effect of oxidative stress injury on the mechanism of vascular smooth muscle cell (VSMC) calciifcation induced by calcium and phosphorus in experimental rats.
Methods: The VSMC calcification was induced by incubating the cells with calcium chloride (CaCl2) andβ-sodium glycerophosphate (β-GP) for 8 days, and the cells were divided into 4 groups: ① Control group, ② Calcification group,③ Calciifcation+H2O2 group, ④ Calciifcation+catalase group. The calcium nodule formation and calcium deposition in VSMC were detected by Alizarin red staining and o-cresolphthalein complexone method, the reactive oxygen species (ROS) was detected by DCFH-DA probe staining and the protein expression of Runx2 was examined by Western blot analysis.
Results: Compared with Control group, Calciifcation group showed the higher ROS production, more calcium nodule and calcium deposition, higher Runx2 protein expression;while compared with Calciifcation group, the above indexes were even higher in Calciifcation+H2O2 group, P<0.05. The ROS production, calcium nodule, calcium deposition and Runx2 protein expression were lower in Calciifcation+catalase group than those in Calciifcation group and Calciifcation+H2O2 group, but still higher than that in Control group. The protein expression of Runx2 was similar between Calciifcation+catalase group and Control group, P>0.05.
Conclusion: CaCl2 andβ-GP treatment may induce VSMC calciifcation via activating ROS-Runx2 signal pathway in experimental rats.