OBJECTIVE:To establish the method for bacterial endotoxin test of Kuzhi injection. METHODS: The test sample dilution method was applied to exclude the interference of the Kuzhi injection in the bacterial endotoxin tachypleus amebocyte lysate(TAL) test. RESULTS: Diluted to 16 times, the Kuzhi injection showed no interference in the bacterial endotoxin TAL test. CONCLUSION: The established method is applicable for the bacterial endotoxins test of the Kuzhi injection.

OBJECTIVE:To establish the method of assaying bacterial endotoxin in cardioplegia solution.METHODS:The tachpleus amebocyte lysate(TAL) test for bacterial endotoxin in cardioplegia solution was performed in accordance with Chinese Pharmacopoeia(2005 Edition,Volume Two).RESULTS:The 8-fold diluents of cardioplegia solution did not interfere with the test for bacterial endotoxins,with Et=0.5 Es～2 Es.CONCLUSION:It is feasible to detect bacterial endotoxins in cardioplegia solution with TAL at a sensitivity of 0.06 EU?mL-1.

Background and purpose:The reason for hepatitis B virus (HBV) with hepatocyte specificity is PreS1 enchased on the hepatitis B virus envelope (HBVE). So HBVE may have a potential application in liver targeting gene transfer. In this study, we investigated whether HBVE has the ability to target liver cancer cells. Methods:HBVE was obtained from the supernatant of Hep G 2.2.15 cells through PEG8000 system and ?-propiolactone method. The pIRS2-EGFP was packed with HBVE and resulted in the product HBVE-GFP. HBVE-GFP was transfected into HepG2, A549, HeLa and FB cells. The green fluorescent protein (GFP) expression was observed under a fluorescent microscope. The rate of GFP positive cells was determined by flow cytometer.Results:The GFP could be observed in the four groups, but the HepG2 group had a higher fluorescent intensity than the other 3 groups. The transfected rate of HepG2 group was (71.35?0.03)% , much higher than other groups(P

Objective To study the validation of fluorescent multiplex microsatellite amplification technique for use in the caseworks of umbilical blood cell transplantation monitor. Methods Six post-transplant recipients of umbilical cord blood cell transplantation were monitored by analyzing microsatellite DNA loci. DNA samples were amplified using a fluorescent labeling primers multiplex amplification system of 15 microsatellites markers, followed by typing on a DNA automated sequencer. Recipients' or donors' microsatellites DNA profiles were compared before and after transplantation. Results All recipients and donors exhibited different DNA profiles. Without reference samples of pre-transplant or donors, the changes of the 15 microsetellites genotypes of the post-transplant recipients still could be analyzed. The recipient type turned to donor type was observed over time. Conclusion Under the condition of using multiplex amplification of the 15 microsatellites to monitor the umbilical blood cell transplantation, reference sample of pre-transplant or donor did not need to be detected simultaneously.

Objective To investigate the STR typing discordance between different typing methods.Methods Genotypes of 13 routine forensic STR loci in DNA samples from 100 individuals were typed by using singleplex polyacrylimide gel electrophoresis silver stain system and Power Plex16 System,respectively.The typing results between these two systems were compared.Results One genotype discordance was observed at D8S1179 locus in a DNA sample.The genotypes was 12/14 in snigleplex system and 12/15 in Power Plex16 System.Conclusion Different STR typing systems may result in different genotyps from the same DNA sample.

Objective:To observe the clinical efficacy and safety of compound Bletilla Striata cream in the treatment of patients with second degree hand rhagadia. Methods: Totally 146 patients with second degree hand rhagadia were randomly divided into the treatment group and the control group with 73 cases in each. The treatment group was treated with compound Bletilla Striata cream, while the control group was treated with urea ointment. The treatment course was 10 days. The time of paln disappearance, keratoder-ma softening and rhagadia healing were compared between the two groups, and the efficacy and adverse drug reactions of the two groups were also observed and compared. Results:After the first treatment course, the curative rate of the treatment group was 94. 5%, which was higher than that of the control group (82. 2%) with statistical significance (P<0. 05). The total effective rate of the two groups was both 100% without statistical significance (P>0. 05). The time of keratoderma softening and rhagadia healing in the treatment group was significant shorter than that in the control group (P<0. 05), while the time of paln disappearance showed no significant difference between the two groups(P>0. 05). Conclusion: Compound Bletilla Striata cream in the treatment of second degree hand rhagadia is effective and safe, which shows good prospect for clinical application.

Objective To observe the degeneration of Brugia malayi in Meriones unguiculatus model. Methods Microfilaria of Brugia malayi derived from Meriones unguiculatus was used to infect Anopheles sinensis . Infective stage larvae (L 3) from mosquito vector were collected and inoculated into abdomen of Meriones unguiculatus. Successive 33 generations of the parasite in the rodent model have been observed. Results Since 1974 when the animal model was established, the parasite has lasted for 33 generations, the positive rate of Meriones unguiculatus with microfilaria gradually reduced from 80% of the 28th generation to 16% of the 32nd generation and finally to 0 at the 33rd generation. Conclusion It became difficult for the larvae of Brugia malayi to develop and/or reproduce in the animal model after multiple inoculations for generations.

Objective: To construct a liver targeting gene transfer vector using hepatitis B virus envelope particles. Methods: Hepatitis B viruses were obtained from the supernatant of HepG 2.2.15 cells by a PEG8000 system and were inactivated by ?-propiolactone to prepare hepatitis B virus envelope. The hepatitis B virus envelope was used to pack 5.3 kb pIRES_2-EGFP to assess their packing ability. Subsequently, the products were studied with ELISA, PCR, SDS-PAGE, and electron microscopy. Finally, the product was used to transfect HepG2 cells and the green fluorescent protein (GFP) expression was observed under a fluorescent microscope. The rate of GFP positive cells was determined by flow cytometer.Results: The acquired hepatitis B virus envelope retained the surface protein HBsAg+pre S_1+pre S_2, but with no virus DNA. The prepared envelope had high packing ability for GFP and the packed GFP had a high transfection rate in HepG2 cell. Conclusion: Hepatitis B virus envelope has been successfully obtained from the supernatant of HepG 2.2.15 cells with a PEG8000 system and ?-propiolactone.

Objective:To observe the transfection efficacy and targeting efficiency of hepatitis B virus envelope(HBVE) as a gene transfer vector for liver cancer cells.Methods:HBVE was obtained from the supernatant of HepG2.2.15 cells with a PEG8000 system and ?-propiolactone.The pIRS2-EGFP was packed with HBVE to obtain HBVE-GFP and was packed with liposome to obtain Liposome-GFP.HBVE-GFP and Liposome-GFP were used to transfect human hepatoblastoma cell line HepG 2 to study the transfection efficiency.HepG 2,A 549,HeLa and FB cells were transfected with HBVE-GFP to appraise the targeting ability of HBVE-GFP.GFP protein expression was observed under a fluorescent microscope and the ratio of GFP positive cells was determined by flow cytometry.Results:(1) Transfection efficiency:The GFP protein was observed in both the liposome group and the HBVE group under the fluorescent microscope;the fluorescent intensity in the HBVE group was 3-4 times that of liposome group as determined by flow cytometry(P