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Objective:To compare the different DNA extraction methods and establish an efficient and sensitive method of DNA extraction from bacteria for PCR and Real-time PCR.Methods:Bacillus coli DNA with the same concentration was extracted by 5 different DNA extraction methods,the Proteinase K method,the alkali-lysis method,the Chelex-100 with NP40 or Triton X-100 and the water-boiling method to determine OD260/280.Bacillus coli DNA with different concentrations wewe extracted to be amplified by PCR and Real-time PCR for compariing the sensitivity.Results:Among these extraction methods,Chelex-100 + NP40 method was with the best purity and sensitivity and offered a higher OD260/280(1.79?0.03).Neither false positive nor false negative was found in PCR and Real-time PCR,The sensitivity was at 10/ml of bacteria concentratio.Conclusion:The DNA extraction based on Chelex-100 + NP40 is effective,specific and sensitive,and it accommodate to PCR and Real-time PCR.

Objective:to study the absorption and absorption characteristics of isomer of propranolol hydrochloride in Caco-2cell monolayers model,which have taken as the most representative model to investigate the absorption mechanism of oral drug in recent years. Methods:Caco-2 cell monolayers model was set up to study transport of isomer of propranolol hydrochloride. RP-HPLC method was applied to analyze isomer of propranolol hydrochloride through precolumn derivatization. Results: Propranolol hydrochloride was absorbed completely,and was a passive diffusion mechanism. There was no chiral selectivity of transport and absorption of them across Caco-2 cell monolayers. Conclusion: Caco-2 cell model was established and the cell integrity was evaluated. The integrity of Caco-2 cell was in good condition,which could be used to study the absorption and transport. RP-HPLC method was established to separated isomer of propranolol hydrochloride within 20 min by precolumn derivatization. The method is convenient, sensitive,accurate and suitable for the analysis of telbivudine.

Objective:To investigate the clinical characteristics and prognostic factors of patients with primary nasal non-Hodgkin's lymphoma(NHL). Methods:The clinical features,diagnosis and treatment of 38 patients with NHL occurring in nasal cavities and paranasal sinuses were retrospectively analyzed. Results:The Ann Arbor stages were:stage Ⅰ,14patients; stage Ⅱ,11 patients;stage Ⅲ,7 patients;stage Ⅳ,6 patients. The most common symptoms at the presentation included nasal obstruction,rhinorrhea and cpistaxis. 11 patients had B symptoms(fever,night sweat,and/or weight loss). The lymphoma cells in all the cases ranged in size from small to large,and necrosis was common. Immunophenotyping were determined in 25cases and all were T cell type. 6 cases with early localized-disease were treated with radiotherapy and 7 had chemotherapy,the remaining 25 cases received combined chemotherapy and radiotherapy. The 5-year overall survival(OS) for the entire group was 78.9%,and those for patients with localized disease(stages Ⅰ and Ⅱ) and those with the advanced disease(stages Ⅲ and Ⅳ) were 92.0% and 53.8%,respectively(?2=11.29, P

Objective:To study whether the glucose tolerance was impaired in the non-hyperglycemic rats treated with strepto-zotocin(STZ).Methods:Male SD rats,aged 10￣12 weeks,were divided into two groups:experimental group,with an intraperitoneal injection of 30 mg/kg(low dose)or 60 mg/kg(high dose)STZ;control group,with injection of equal dose acidified saline.1￣12 weeks after injection,the intraperitoneal glucose tolerance tes(tIPGTT)was performed in the non-hyperglycemic rats(fasting blood glucose

Objective:To investigate the role of proteinase-activated receptor 1 and 2(PAR1,PARs)in regulation ofthe expression oftype I/Ⅲ collagen protein of hepatic stellate cell isolated and cultured from SD rats liver.Methods:Hepatic stellate cell isolated and cultured from SD rats livers with a modification technique of Friedman and centrifugation of Nycodenz Gradient.Expression of mRNA for PAR1, PAR2,Ⅰcollagen and Ⅲ collagen were examined by reverse transcription polymerase chain reaction(RT-PCR)and protein examined by immunocytochemistry,observated characteristic cell shape,and examined expression of hexadecanoic acid(HA)and laminin(LN)of supernatant from culturesd HSC.Results:In the course of culturing rat HSC,vitamin A autofluorescence and lipid droplet were decreased gradually or lost following HSC activation,the cells morphology changed to stellate cells and cells proliferation activated.Ⅰ/Ⅲ collagen protein expressions were also increased,the expression of HA,LN,PAR1 and PARs paralleled with the expression ofⅠ/Ⅲ collagen protein and correlated with activation and proliferation of HSC.Conclusion:The expression of PARs correlated with activation and proliferation of HSC,could promote the expression ofⅠ/Ⅲ collagen protein,HA and LN.PARs might play an important role in the mechanisms of hepatic fibrosis.

The oocyte cryopreservation is more promising than embryo freezing in clinical applications,preservation of female fertility,legal and ethical aspects in this decade.The redundant oocytes retrievedfrom ⅣF cycles can potentially donate oocytes,which have unique advantages in economy and feasibili-ty.We have achieved success in embryo freezing,but just obtained poorer results in oocyte cryopreserva-tion which was mainly because of the low rates of survival,fertilization,and cleavage.The character ofthe plasma membrane and the time of cortical granules present at the metaphase of meiosis Ⅱ with thespindle system consist of the major difference between oocytes and embryos.Moreover,the oocytes shouldbe fertilized by sperm at the appropriate time.We used a refined slow freezing method to improve the sur-vival rate and increased sucrose concentration to dehydrate oocytes.Vitrification was another approach toprevent harms.Besides,intracytoplasmic sperm injection was used to overcome possible zona hardeningafter the release of cortical granules.Oocytes after cryopreservation showed serious disturbances of the mi-crotubules immediately after thawing as they were vulnerable to the thermal changes and easy to depoly-merize.Fertilization of these oocytes with disorganized spindles led to chromosomal aneuploidy,digyny,and degradation of the cleavage rate.We can improve cryopreserved oocytes to normal fertilization anddevelopment by appropriate incubation and timing of insemination,as the microtubules repolymerize in atime dependent way after incubation which is compatible with recovery of the spindles.With the improve-ment of survival,fertilization,and development,oocyte cryopreservation will play an imperative role.

Heme oxygenase(HO) is the rate-limiting enzyme in the degradation of heme.Three HO isoforms have been identified:HO-1,HO-2 and HO-3.The substances of heme degradation,carbon monoxide(CO),biliverdin and free iron(Fe~(2+)),have an important function in anti-oxidant and protecting cells.And the anti-oxidant function of induction HO-1 expression is mediated in the signal transduction of HO-1 gene expression and the promoter of HO-1 gene in the cells.So the new way is provided in the relevent liver-injury which is caused in anti-oxidant stress or non-oxidant stress.

Objective: To explore the effect of sequential external counterpulsation on sudden deafness.Methods: All the patients were treated with sequential external counterpulsation instrument produced by Guangzhou South-China Medical Instrument Company.Results: After treatment,the hearing threshold of 60 cases was all improved,with an increase from 5 to 53 db,and with an average threshold shift of 29?8.7db.86.7% of cases acquired satisfied therapeutic effects except 8 cases whose hearing threshold increased by less than 15db.Conclusion: the results showed that,the treatment of sudden deafness with Sequential External Counterpulsation has good therapeutic effects.Earlier and longer treatment means better effect

Objective To determine the content of hesperidin in Weifukang capsules by HPLC,and to establish the quality standard of Weifukang.Methods HPLC method was performed on C18 column(4.6 ?200 mm,5 ?m).The mobile phase consisted of acetonitrile-sodium dihydrogen phosphate(0.05 mol/L)-phosphoric acid(200 ∶800 ∶0.5).The flow rate was 1.0 mL/min,column temperature was 25 ℃,and the detection wavelength was 283 nm.Results Hesperidin was linear at the range of 0.38 ?g～1.90 ?g(r=0.999 2)the mean recovery rate was 97.88 %,and RSD was 1.69 %.Conclusion This method is effective and reproducible and provides a basis for the quality control of Weifukang capsules.