TNF? mRNA in human bronchoalveolar lavage(BAL)cells was quantitated by a olymerase chain reaction(PCR)using a cRNA internal standard,cRNA molecules were in vitro transcribed from pAW108 plasmid in which the sequences of upstream primer and omolementary sequences of downstream primer of TNF? were inserted.The cRNA and mRNA extracted from BAL cells were mixed and reverse-transcribed into cDNA.The resultant cDNA mixture was 1:3 diluted and amplified by PCR using TNF? specific primers.~(32)p-labeled upstream primers were included in the PCR reaction,cDNA fragments amplified was run on a 3% agarose gel.The radioactivity of positive bands was determined in a scintillation ounter.After plotting variable template concentrations of the internal standard pAW108 cRNA and the number of BAL cells against the radioactivity of their PCR products,the levels of TNF? mRNA in BAL cells were quantitated by comparision to those of cRNA internal standard.

OBJECTIVE:To discuss the necessities and countermeasures of pharmaceutical care in the rural areas of China.METHODS:The existing difficulties of pharmaceutical care in the rural areas of China were analyzed realistically.RESULTS&CONCLUSION:In order to successfully carry out pharmaceutical care,firstly the state food and drug adminis-tration must strengthen the supervision,construct the supply network of drugs and ensure the quality of medication;secondly the pharmacists'ability must be enhanced and pharmacists'quantity increased through various of training and education;thirdly the consultation service should be carried out by specially assigned persons.Only in this way can we improve the quality of pharmaceutical care and ensure the secure,efficient and economic drug use.

Objective To study the potential effect of gene-environment interaction between glutathione S-transferase M1 (GSTM1) and serum organochlorines residues on the risk of breast cancer in women, in China. Methods Seventy newly pathologically diagnosed female patients with breast cancer from September 2006 to October 2007 were selected as the cases from five large hospitals in Tangshan. The controls were identified at the same hospital as cases. 1∶1 matched case-control study. Between the cases and controls, the difference of age was not over two years and the residence was similar. The organochlorine residues levels in the serum were measured by gas chromatography (GC). Genotypes of GSTM1 polymorphisms were analyzed by multiplex allele-specific polymerase chain reaction (PCR). Interaction indexes (?) were calculated to determine the type of gene-environment interaction. Results After confounding factors adjusted, the result showed that interaction existed in genetic polymorphisms of GSTM1 and DDT, HCH residues, and interaction indexes (?) value were 1.237 and 1.379. Conclusion GSTM1 genetic polymorphisms and DDT, HCH may present an interaction in the development of breast cancer.

Abstract Objective:To construct bmcella melitensis Br.melitensis genomic DNA expression library and study its immune effect.Meth-ods: Hind Ⅲ digested Br.melitensis genomic DNA fragments were cloned into pSV-?-Gal plasmds.The recombinant plasmids were transformedinto JM109 host bacteria. After large-scale isolation, the recombinant plasmids were inoculated into the quadriceps muscle of mice. In the 9thday after third boost immunization .agglutination antibodies against Br. melitensis and lymphocyte mitogenic ability of mice were determined. Re-sults : Expression library nucleic acid vaccine can induce the production of agglutination antibodies against Br. melitensis and stimulate the lym-phocyte porliferation primed by ConA. Conclusion:The genomic DNA expression library may provide a new vaccine against Br. melitensis infec-tion.

Using human acidic fibroblast growth factor Specific primers we tested the technique of screening cDNA library by a DNA polymerase chain reaction.With the method,we have suc-cessfully isolated encoding region cDNA of human interleukin 8 from both human dermal fibrob-last and PMA stimulated U937 cell cDNA libraries.

Objective:To investigate the protective effect and potential mechanism of cordycepin on renal proximal tubular cells injury induced by lipopolysaccharide (LPS).Methods:Renal proximal tubular cells NRK-52E were incubated on a cell culture plated at a density of 1×10 5/mL for experiment, then divided into control group (Ctrl group), LPS group (cells were stimulated with 1 mg/L LPS), 10 μmol/L or 20 μmol/L cordycep in intervention groups (LPS+C 10 group and LPS+C 20 group). Cell viability was measured using cell counting kit-8 (CCK-8) reagent. The level of intracellular reactive oxygen species (ROS) was detected by 2',7'-dichlorofluorescin diacetate (DCFH-DA) staining. The protein expressions of inflammatory factors intercellular cell adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), interleukin-1β (IL-1β), and nuclear factor-κB (NF-κB) were detected by Western blotting. Results:Compared with the Ctrl group, LPS significantly inhibited NRK-52E cell viability, increased intracellular ROS, and up-regulated the expressions of ICAM-1, VCAM-1, IL-1β and NF-κB. Compared with LPS group, after treated with 10 μmol/L or 20 μmol/L cordycepin, NRK-52E cell viability was significantly increased (Ctrl group as 1: 0.717±0.017, 0.916±0.036 vs. 0.554±0.046) and intracellular ROS level was significantly decreased (Ctrl group as 1: 1.527±0.165, 1.098±0.168 vs. 2.543±0.127), meanwhile the expressions of ICAM-1, VCAM-1, IL-1β and NF-κB were significantly down-regulated [Ctrl group as 1, ICAM-1/GAPDH: 2.364±0.097, 1.561±0.074 vs. 3.101±0.121; VCAM-1/GAPDH: 2.866±0.135, 1.920±0.098 vs. 4.170±0.119; IL-1β/GAPDH: 2.358±0.107, 1.563±0.179 vs. 3.301±0.210; phosphorylation NF-κB p65 (NF-κB p-p65)/GAPDH: 2.559±0.166, 1.596±0.148 vs. 3.183±0.098], the differences were statistically significant (all P < 0.05). Compared with the LPS+C 10 group, the cell activity of LPS+C 20 group was more significant (0.916±0.036 vs. 0.717±0.017, P < 0.01), and the expressions of ICAM-1, VCAM-1, IL-1β, NF-κB were down-regulated more significantly (ICAM-1/GAPDH: 1.561±0.074 vs. 2.364±0.097, VCAM-1/GAPDH: 1.920±0.098 vs. 2.866±0.135, IL-1β/GAPDH: 1.563±0.179 vs. 2.358±0.107, NF-κB p-p65/GAPDH: 1.596±0.148 vs. 2.559±0.166, all P < 0.05).Conclusion:Cordycepin could significantly increase the survival rate of NRK-52E cells, reduce intracellular ROS level, and inhibit inflammation, and the anti-inflammation effect can be related with NF-κB pathway.

AIM:There are some reports about the treatment of spinal cord injury with neural stem cells(NSCs) transplantation,but the ideas about the transplantation time and patterns and detection indexes are still different.In this study,the effects of cryopreserved NSC transplantation on the axonal regeneration after the spinal cord injury in rats were observed.METHODS:The experiment was carried out in the experimental animal center of China Medical University from June 2005 to June 2006.①Thirty-six adult Wistar rats,either male or female and 250-300 g,were provided by the experimental animal department of China Medical University.Neural stem cells were isolated from 10 neonate rats and cultured.All treatments for animals were accorded with the animal ethical criteria.②NSCs at logarithmic phase were cryopreserved at-70 ℃ for 2 weeks and labeled with 5-bromodeoxyuridine(Brdu) after rewarming.After the models of spinal cord injury were established,the NSCs were transplanted into the injured site immediately.Thirty-six rats were randomly divided into NSC transplantation group,DMEM solution group,and control group.③NSC survival and migration were detected by immunohistochemistry,and the reconstruction of spinal cord were detected by horseradish peroxidase(HRP) staining.RESULTS:Of the 36 spinal injured model rats,4 died of overdose of anesthesia,and 5 died of infection,but all were supplemented.Brdu-labeled positive NSCs were detected in the injured spinal cord after transplantation on the 7th day,and increased on the 14th day,then gradually decreased since the 28th day till disappeared.The number of HRP positive cells in transplantation group was significantly higher than DMEM solution.CONCLUSION:Cryopreserved NSCs can survive in the injured site after transplantation and promote the reconstruction of axoplasm pathway of injured spinal cord.

In recent years,the humanistic quality education in vast number of medical colleges and universities shows a low level of wandering state.This in not only related with the poor condition of current humanistic quality course,but also related with the diametrically opposed humanistic quality education and medical professional education and lack of organic integration.To improve the medical colleges' educational level is urgent to establish the concept of holographic education of great medicine,great education and great culture,improve the teaching quality of current humanistic quality education,actively explore the deep integration of humanistic quality education and medical professional education,improve the humanistic quality and nurturing ability of teachers staff in medical universities and colleges,and so on.

An overall improvement of teaching quality is the core task of the reform and the development of higher education. Based on the analysis of the main factors of affecting the teaching quality in universities and combined with the implementation of “views on improving the overall qual-ity of higher education”by the Education Ministry, countermeasures for improving the teaching quality of undergraduates have been proposed such as increasing higher education funds investment to im-prove the condition of running a school, paying attention to the construction of teaching management staff to improve the level of teaching management, establishing Teacher Development Research Center to consolidate the fundamental position of undergraduate teaching , utilizing the high-quality social resources to implement the collaborative education innovation, and improving the personnel training system to strengthen the teaching quality standard .