Objective To establish a specific fluorescence quantitative method for determining the mRNA expression of Toll-like receptor3(TLR3)in peripheral blood mononuclear cells(PBMCs).Methods Using the Beacon Designer 2.1 software,specific primers and Taqman-MGB probe were designed.The plasmid pMD18-T-TLR3 was constructed as calibrator and the amplified fragment was obtained by reverse- transcript-PCR(RT-PCR).RNA quantification based on cycle threshold values(Ct)was used to establish the standard curve.According to which,the TLR3 mRNA levels in 30 normal individuals,20 patients with primary biliary cirrhosis(PBC)and 20 ones with chronic liver cirrhosis induced by HBV were calculated automatically by software after the fluorescence of PCR product was detected continuously during amplification.Results The linear detection range of the assay for TLR3 gene and ?-actin was 10~2-10~8(r= -0.9974)and 10~3～10~8(r=-0.9984),respectively.The coefficient of variation of both intra-and inter- assay reproducibility for high concentration sample were 6.7% and 8.7%,respectively,and those for low concentration sample were 12.3% and 14.0%.The TLR3 mRNA expression level ranges from 3.46?10~2- 4.51?10~3 copies/?g RNA,4.92?10~2-1.42?10~4 copies/?g RNA and 2.58?10~2-7.17?10~3 copies/?g RNA for 30 healthy individuals,20 PBC patients and 20 ones with chronic liver cirrhosis induced by HBV, respectively.Conclusion We have successfully set up a FQ-RT-PCR method for detecting TLR3 mRNA, which may be used as an excellent tool for the clinic and basic study on the expression of TLR3 gene.

The leakage of genetically engineered microorganism(GEM) at the beginning of operation was the important content for ecological risk assessment when GEM was inoculated in a membrane bioreactor(MBR) for bioaugmentation. The effects of different operating conditions on leaking density and intercepting efficiency of GEM were investigated in a submerged microfiltration MBR at the beginning of operation. The interception characteristics were also discussed. The results showed that different operating conditions had different influences on intercepting efficiency:higher sludge concentration was profit for interception,while higher aeration intensity and flux had opposite effects on interception. When the inoculated density of GEM was 1.0?1010 CFU/mL,the leaking densities varied from 1.0?102 CFU/mL to 2.5?102 CFU/mL under different operating conditions at the beginning of operation and the maximum intercepting efficiency could be higher than 8 lg. The main factors determining intercepting efficiency at the beginning of operation were membrane module interception,sludge adsorption as well as suspended GEM transfer inhibition and their contribution shares under certain conditions were 82.3%,14.9% and 2.8%,respectively. Gel layer formation during MBR stable operation was helpful to increase intercepting efficiency. The contribution shares for GEM interception of membrane module,sludge and gel layer were 75.3%,10.7% and 14.0%,respectively,under certain conditions.

According to the characteristics of sludge samples from full-scale wastewater treatment bioreac-tors, the essential total DNA extraction method for most environmental samples, lysozyme-SDS-phenol/ chloroform method, was modified to improve sample pretreatment, intensify cell lysis and enhance the effi-ciency of impurity removal. Obtain a general total DNA extraction method for industrial sludge samples. Such a method was applied for total DNA extraction of sludge samples from several running full-scale an- aerobic or aerobic bioreactors in Shijiazhuang, China. The results indicated that the modified method was suitable for all the sludge samples in this study, showing the satisfying generality. The extracted total DNA of all sludge samples were pure, with about 1.8 of A260/ A280 ratio. The method was also efficient; with average total DNA yield of over 0.7 mg/g and maximum yield of 0.85 mg/g. Moreover, all the extracted to- tal DNA samples could serve as templates directly to amplify 16S rDNA by PCR. The PCR products could be separated well by denaturing gradient gel electrophoresis (DGGE) and the DGGE band patterns were clear enough to be used for further analysis. All these facts indicated that the total DNA extraction method provided in this study could meet the requirements of sludge samples research, from full-scale wastewater treatment bioreactors, using molecular biology technologies.

The effect of low temperature storage on dormancy breaking, sprouting and growth after planting of Tulipa edulis was studied. The results showed that starch content and activity of amylases significantly decreased during 10 weeks of cold storage, soluble protein content raised at first then decreased, and the peak appeared at the 6th week. However, total soluble sugar content which in- creased slowly at first than rose sharply and reducing sugar content increased during the storage duration. The bulbs with cold storage treatment rooted in the 6th week, which was about 2 weeks earlier than room temperature storage, but there were less new roots in the late period of storage. After stored at a low temperature, bud lengths were longer than that with room temperature treatment. Cold storage treatment could promote earlier emergence, shorten germination time, prolong growth period and improve the yield of bulb, but rarely affect the emergence rate. It was not beneficial to flowering and fruiting. The results indicated that 6-8 weeks of cold storage was deemed to be the key period of dormancy breaking preliminary.
Cold Temperature ; Plant Dormancy ; Plant Roots ; chemistry ; growth & development ; physiology ; Tulipa ; chemistry ; growth & development ; physiology

The study was conducted to explore the response of growth and yield of Tulipa edulis to flower bud removal and artificial pollination. And flower bud removal and artificial pollination were carried out in the squaring period and bloom stage respectively. The morphological index and biomass indicators were determined and the yield was counted in harvest time. Result showed that flower bud removal was beneficial to the growth of T. edulis, resulting in increasing growth index, biomass as well as the yield of bulb. The diameter and dry weight of T. edulis fruit by artificial pollination were increased significantly compared with the control. Seed setting percentage increased to 100%, and the number of seed as well as the single grain weight increased by 69.03% and 16.48%, respectively, which did not significantly affect the bulb production. In conclusion, Flower bud removal treatment accelerates bulb biomass increase, so as to improve its yield. Artificial pollination raised significantly seed setting percentage, seed number as well as the single grain weight.
Biomass ; Botany ; methods ; Flowers ; growth & development ; physiology ; Pollen ; growth & development ; physiology ; Pollination ; Tulipa ; growth & development ; physiology

Clinical Laboratory Techniques ; Hepatitis, Autoimmune ; diagnosis ; Humans

Derris eriocarpa, a traditional Chinese medicine belonging to the family of Leguminosae, is widely distributed mainly over Yunnan, Guangxi and Guizhou of China. Modern pharmacological researches on this herb showed that it had extensive bioactivities, such as promoting urination, removing dampness and cough and reducing inspissated mucus and other biological activities. The extensive studies on the chemical constituents of this plant have resulted in the isolation of triterpenoids, steroids, fatty acid and others, but the flavone compounds haven't reported before. In our further research on the ethyl acetate of this plant, nine flavone compounds were obtained by column chromatography on silica gel, Sephadex LH-20, semi-prep HPLC, polyamide column chromatography and recrystallization for separation and purification. The structures were determined on the basis of extensive spectroscopic analysis, including MS, NMR experiments and comparison with spectroscopic data in the literature, respectively, as diosmetin (1), 3, 3'-di-O-methylquercetin (2), afromosin (3), 6, 3'-dihydroxy-7, 4'-dimethoxyisoflavone (4), odoratin (5), 7, 3'-dihydroxy-8, 4'-dimethoxyisoflavone (6), 6, 4'-dihydroxy-7, 3'-dimethoxyisoflavone (7), 5, 7, 4'-trihydroxy-3, 3', 5'-trimethoxyflavone (8), and alpinumisoflavone (9). All these compounds were isolated from Derris eriocarpa How for the first time. And the in vitro assays showed that compound 2 possessed moderate inhibitory activity against human cancer cells K562 and HEL.
Derris ; chemistry ; Flavonoids ; chemistry ; isolation & purification ; pharmacology ; Humans ; K562 Cells

OBJECTIVETo investigate the oxidative damage to lung tissue and peripherial blood in PM2.5-treated rats.

METHODSPM2.5 samples were collected using an auto-sampling instrument in summer and winter. Treated samples were endotracheally instilled into rats. Activity of reduced glutathione peroxidase (GSH-Px) and concentration of malondialdehyde (MDA) were used as oxidative damage biomarkers of lung tissue and peripheral blood detected with the biochemical method. DNA migration length (microm) and rate of tail were used as DNA damage biomarkers of lung tissue and peripheral blood detected with the biochemical method.

RESULTSThe activity of GSH-Px and the concentration of MDA in lung tissue significantly decreased after exposure to PM2.5 for 7-14 days. In peripheral blood, the concentration of MDA decreased, but the activity of GSH-Px increased 7 and 14 days after experiments. The two indicators had a dose-effect relation and similar changing tendency in lung tissue and peripheral blood. The DNA migration length (microm) and rate of tail in lung tissue and peripheral blood significantly increased 7 and 14 days after exposure to PM2.5. The two indicators had a dose-effect relation and similar changing tendency in lung tissue and peripheral blood.

CONCLUSIONPM2.5 has a definite oxidative effect on lung tissue and peripheral blood. The activity of GSH-Px and the concentration of MDA are valuable biomarkers of oxidative lung tissue damage induced by PM2.5. The DNA migration length (microm) and rate of tail are simple and valuable biomarkers of PM2.5-induced DNA damage in lung tissues and peripheral blood. The degree of DNA damage in peripheral blood can predict the degree of DNA damage in lung tissue.

Animals ; DNA Damage ; drug effects ; Drug Administration Routes ; Drug Administration Schedule ; Lung ; drug effects ; pathology ; Lung Diseases ; blood ; chemically induced ; pathology ; Male ; Oxidative Stress ; Particle Size ; Particulate Matter ; administration & dosage ; toxicity ; Rats ; Rats, Wistar ; Seasons

Objective To find out the status of brick-tea type fluorosis in the epidemic areas.Methods Based on "Scheme for Epidemiological Brick-tea Type Fluorosis in Sichuan Province",ten counties were selected in Sichuan brick-tea areas and ten towns were selected in every county,then the epidemicologic survey was performed in children of 8～12 year-old and adults aged above 20 years old.Results 5044 children and 4053 adults were selected from brick-tea areas.The rates of dental fluorosis in children and adults were 55.69%(2809/5044)and 60.41%(4053/6709)respectively.The dental fluorosis was mainly of mild damage.The skeletal fluorosis found in X-ray film was 44.64%(167/1241)and in clinical examination,38.94%(3883/9973).The levels of urine fluoride in children and adults were 1.88 and 2.78 mg/L.The level of urine fluoride was not differenet among children of different age,but in adults it was higher in the elder than the younger.The level of fluoride in urine was related to the severeness of skeletal fluorosis(r=0.74).The detective rates of skeletal fluorosis in agricuIture,pasturing,and agriculture-pasturing areaswere 31.70%(1369/4318),50.04%(1228/2454),and 40.17%(1286/3201),respectively.The X-ray detecting rates of skeletal fluorosis in men and wonlen were 49.57%(229/462)and 41.72%(325/779) respectively(χ2=11.72,P＜0.05).Conclusion The prevalence of brick-tea type fluorosis is very serious in the regions studied.

OBJECTIVETo study the effects of prophylactic intra-iliac and hepatic arterial infusion chemotherapy on pelvic recurrence and liver metastasis after radical resection for rectal cancer.

METHODSEighty-four rectal cancer patients,undergone radical resection on Dukes stage B or C,were randomly assigned to postoperative intra-iliac and hepatic arterial infusion chemotherapy group(group I) and routine vein chemotherapy group(group II). Five-year survival and recurrence rates were compared between the two groups.

RESULTSAmong the 84 rectal cancer patients with radical resection, the 5-year liver metastasis and pelvic recurrence rates were 30.2% (13/43) and 18.6% (8/43) respectively in group II, 17.1% (7/41) and 9.8% (4/41) in group I, the difference was significant between 2 groups (chi(2)=4.31, P<0.05). The mean tumor-free survival time was 26.2 months in group I and 15.8 months in group II (t=5.05, P<0.01), the difference was significant (t=5.05, P<0.01). The five-year survival rate in group I (65.9%) was significantly higher than that in group II (56.5%) (u=8.86, P<0.01). Cox multivariate analysis showed that, compared with those in group II, the relative risks of pelvic recurrence and liver metastasis in group I decreased 20% (coefficient of relative risk: 0.7959), and the five-year mortality also decreased 20% (coefficient of relative risk: 0.8034).

CONCLUSIONProphylactic intra-iliac and hepatic arterial infusion chemotherapy can reduce the rates of pelvic recurrence and liver metastasis after radical resection of rectal cancer.

Adult ; Chemotherapy, Adjuvant ; Chemotherapy, Cancer, Regional Perfusion ; Female ; Hepatic Artery ; Humans ; Iliac Artery ; Liver Neoplasms ; prevention & control ; secondary ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; prevention & control ; Pelvic Neoplasms ; prevention & control ; secondary ; Pelvis ; pathology ; Rectal Neoplasms ; drug therapy ; pathology ; Survival Rate