Objective To establish clonal human embryonic stem cell lines and investigate their biological characteristics. Methods Cells were derived from one inner cell mass of human blastocyst, multiplied for 20 passages, and then dissociated into single cell suspension by digestion with 0.5% trypsin. Single cell was picked up and plated into individual well of a 96-well plate containing feeder-layers directly under a dissection microscope. The outgrowth clones were passed by treatment with collagenase. Surface markers were detected by cytochemistry and histoimmunochemistry. Karyotypes were tested using standard G-banding techniques. The pluripotency was analyzed by inoculating cells into severe combined immunodeficient (SCID) mice. Results Two clonal human embryonic stem cell lines were established. Cells of these two lines possess the characteristics and differentiating potencies: normal 46 XX karyotypes; expressing a series of surface markers such as: alkaline phosphotase, stage-specific embryonic antigen (SSEA)-4, tumor recognition antigen (TRA)-1-60, TRA-1-81 etc; and forming teratomas comprising derivatives of three embryonic germ layers such as neural tissue, cartilage, squamous epithelium and columlar epithelium when injected into SCID mice. Conclusions The two single cell-cloned human embryonic stem (hES) cell lines were derived in our laboratory. The cells possess stable biological characteristics of undifferentiated hES cells.

OBJECTIVE: To establish the HPLC method for the determination of chloramphenicol and dapsone in compound chloramphenicol cream. METHODS: The analytical column was Hypersil ODS and the mobile phase consisted of 0.1%sodium heptanesulfonate solution (a mixture of 0.1% sodium heptanesulfonate 500 mL and dimethylformamide 5 mL plus acetic acid glacial 0.5 mL)-acetonitrile (75∶25) at a flow rate of 1.0 mL?min-1. The detection wavelength was 272 nm and the sample size was 20 ?L. RESULTS: The linear ranges of chloramphenicol and dapsone were 25.59～153.54 ?g?mL-1 (r=0.999 9) and 12.57～75.42 ?g?mL-1 (r=0.999 9), respectively. The average recovery rates were 100.62% (RSD=0.92%) and 100.93% (RSD=1.22%), respectively. CONCLUSION: The method is simple and accurate and can be used for the quality control of compound chloramphenicol cream.

Objective:To study the effect of cervical canal mucosa dng excision on cervical columnar eversion in Loop electrosurgical excision procedure(LEEP).Methods:A prospective randomized control trail was performed in 125 cervical intraepithelial neoplasia(CIN)patients.Cervix and cervical canal were conically excised with triangle electrode in 62 patients in the control group;in the study group,a small ring electrode was put into the cervical canal and the cervical canal mucosa was excised about 0.5～0.8cm,after excision as the control group.Follow-up was performed postoperatively in 1,3,6 months respectively.The operation time,the bleeding volume,the rate of cervical columnar eversion and cervical adhesion or stenosis were compared between the two groups.Results:The rate of cervical columnar eversion in the study group (1/63,1.59%)was lower than that in the control group(9/62,14.52%).There was significantly statistical difference between them(P=0.008).There was no statistical differences between them in the operation time,the bleeding volume,and the rate of cervical adhesion or stenosis (P>0.05).Conclusions:The cervical canal mucosa ring excision in LEEP can effectively prevent postoperative cervical columnar eversion.

Objective To understand the spectroscopic property of AO-H2SO4-KBrO3-naphthol system and the enhanced mechanism of resonance light scattering(RLS),and to develop a method for the determination of trace naphthols in urine.Methods In the dilute H2SO4 medium,naphthols could react with KBrO3 and AO to form ion-association complexes,which produced a new RLS spectrum and resulted in the great enhancement of RLS.The characteristics of RLS spectrum,three-dimensions fluorescence spectrum,absorption spectrum and fluorescence spectrum and the optimum conditions of reaction were studied.Results The enhanced intensity of RLS was 468 nm.The linear range was at 1.41?10-7-2.80?10-5 mol/L for ?-naphthol,1.28?10-7-3.00?10-5 mol/L for ?-naphthol.The relative coefficient and the limits were 0.999 6 and 0.422?10-7 mol/L,0.999 3 and 0.385?10-7 mol/L for ?-naphthol and ?-naphthol,respectively.The urine samples analysis of the relative standard deviation was 4.8%-7.3% and the average recovery rate was 90.7%(n=6).Conclusion This method is sensitive,simple,rapid and applicable to the determination of trace naphthols in human urine.

Objective To establish a method to determine six kinds of PAEs in human serum simultaneously,and on this basis to understand the level of PAEs in human body.Methods Ultrasonic extraction,gas chromatography-mass spectrometry-selected ion monitoring(GC-MS-SIM) detection and quantitative analysis based on internal standard were used to detect six kinds of PAEs in human serum simultaneously.Finally this method was used in fifty human serum samples.Results The results showed that this method had a good linear relation in the range from 10 to 1 000 ng/ml,and the correlation coefficients of standard curve were higher than 0.999.The average recovery rate and the relative standard deviation(RSD,n=6) of the target compounds in standard-addition blank was 94.5% and 4.4% respectively,the lower limits of detection for DEP,DMP,DBP,BBP,DEHP,DNOP were 0.31,0.85,0.90,0.63,0.88,0.41 ng/ml respectively,and the recovery rates of all samples ranged from 67.98% to 110.8%.Conclusion This method has good recovery rate,reproducibility,lower limit of detection and can be used for the determination of many kinds of PAEs substances in human serum.

Purpose　The aim is to study the transduction of m urine stem cell factor(SCF) into umbilical blood cells by LipofectinRMmediated and expression in them.Methods　Murine SCF cDNA encoding extra cellu lar domain was isolated using PCR from plasmid pRC/CMV(containing SCF gene), and then recombined into the expression vector pcDNA3,and transferred into enrichme nt cultural umbilical blood cells. Semi-quantitative RT-PCR was used to examin e the expression on mRNA level.And the effects of supernatant of transfected umb ilical blood cells were investigated alone or in coordinate with GM-CSF by colo ny formation test of human bone marrow cells in semisolid culture.Results 　SCF mRNA was expressed in transfected umbilical blood cells.The su pernat ant of transfected umbilical blood cells could increase the number of CFU-GM, s ynergizing with GM-CSF.Conclusion　The supernatant of umbil ical b lood cells transfected with vector containing mSCF gene can stimulate the colony formation of human bone marrow cells in combination with GM-CSF.

Based on theories of career guidance and refined management,this paper introduced the investigation and practice of refined career planning and guidance for pharmacy graduate students in Peking University,and puts forward some ideas to improve the career planning and guidance in the future.

Objective: To study long time aerobic training effect on mitochondrial DNA (mtDNA) content and activities of mitochondrial respiratory chain complexes in the myocardium and brain tissues of aged rats. Method: Forty-five rats were randomly divided into three groups Group A(n=15)younger rats did not received training; Group B (n=15) old rats did not received training; Group C (n=15) old rats received gradually training for 90 days. The mtDNA content were determined by methods of Yah erc, the activity of mitochondrial respiratory chain complexes in myocardium and brain tissues were determined by methods of Wu etc. Result: The mitochondrial DNA content in myocardium and brain tissues significantly increased (P<0.01) and the activity of mitochondrial respiratory chain complexes in myocardium and brain tissues significantly decreased (P<0.05-0.01) in the aging control group as compared with those in the young group. The mitochondrial DNA content in myocardium significantly decreased (P< 0.05), while there was no significant difference in brain. The activity of mitochondrial respiratory chain complexes in myocardium and brain tissues significantly incroased(P<0.05-0.01) in the aging training group as compared with those in the aging control group. Conclusion: the long time aerobic training could decline the mitochondrial DNA content and increase the activity of mitochondrial respiratory chain complexes in myocardium and brain tissues in aging rats.

The target organs of the most common complications in thoracic radiotherapy mainly contain esophagus,lung and cardiovascular system.Studies of influences such as the related physical dose factors and combined chemotherapy and so on,susceptible biomarks,protective factors,become the study focus,which provide the light for clinical prevention and therapy.

Objective To observe the clinical efficacy of Lao Shi Jiu (moxibustion at a group of ten acupoints) plus Wu Zhu Xiang Sha Liu Jun decoction in treating chronic gastritis due to deficient cold in spleen-stomach. Method Ninety patients with chronic gastritis due to deficient cold in spleen-stomach were randomized into a moxibustion-medication group and a Western medication group by using the random number table, 45 cases each. The moxibustion-medication group was intervened by Lao Shi Jiu plus Wu Zhu Xiang Sha Liu Jun decoction, while the Western medication group was prescribed with conventional medications. The clinical efficacy and changes in symptoms were observed after the intervention. Result The total effective rates were respectively 88.9% and 75.6% in the moxibustion-medication group and Western medication group, and the rate in the moxibustion-medication group was significantly higher than that in the Western medication group (P<0.05); the gastroscopy-based total effective rates were respectively 91.1% and 77.8% in the moxibustion-medication group and Western medication group, indicating that the moxibustion-medication group was superior to the Western medication group (P<0.05); the improvement of stomachache in the moxibustion-medication group was more significant than that in the Western medication group (P<0.05). Conclusion The moxibustion-medication group is superior to the Western medication group in comparing the clinical efficacy and improvement of symptoms in treating chronic gastritis due to deficient cold in spleen-stomach.