OBJECTIVE:To optimize the extraction of effective ingredients and purification for crude glycyrrhizic acid and liquiritin from Radix et Rhizoma Glycyrrhizae with different methods. METHODS:The contents and extraction percentages of glycyrrhizic acid and liquiritin were used as indexes to optimize the extraction solvent. Then the extraction process was optimized with orthogonal experiment taking the concentration of ammonium ethanol,the amount of solvent and the extracting times as indexes. Crude glycyrrhizic acid was purified with the method of recrystallization and crude liquiritin was purified with organic solvent method.RESULTS:The optimal extracting solvent was ammonium ethanol. The optimum extraction conditions were as follows:extracting the solvent 4 times(2 h/time) by adding 5 times volume of 60% ethanol(containing 0.3% ammonia water). The purity of glycyrrhizic acid was over 85% with total extraction rate at 43%,and content of liquiritin was above 15% with its total extraction rate at 67%. CONCLUSION:The method is simple and feasible and from which glycyrrhizic acid and liquiritin was been obtained,Radix et Rhizoma Glycyrrhizae can broaden the scope of the application.

BACKGROUND:To improve the survival rate of transplanted hematopoietic stem cells, dynamic monitoring of plasma content of human cytomegalovirus (HCMV) and rapid screening of early active HCMV infection in hematopoietic stem celltranplantation recipients, thus to guide the clinical medication, is preferred. OBJECTIVE:To evaluate the usefulness of fluorescence quantitative PCR assay for early detection of HCMV activation after hematopoietic stem celltransplantation. METHODS:Real-time fluorescence quantitative PCR assay was applied for HCMV monitoring in 656 blood samples from 41 hematopoietic stem celltranplantation recipients and 60 control blood samples. RESULTS AND CONCLUSION:In 656 blood samples, 96 samples were positive, and the HCMV DNA copies ranged from 5.0×102 to 1.0×107 IU/mL. Timely initiation of therapy resulted in the rapid clearance of DNA-viraemia but it recurrenced in short time by drug-resistent. Two cases from 12 postive recipients died from HCMV infection. The quantitative detection of HCMV DNA by real-time PCR is a rapid method for monitoring HCMV infection and the early diagnosis in patients after hematopoietic stem celltransplantation.

OBJECTIVE:To investigate the use of antibacterials and to evaluate the distribution and drugs resistance of pathogenic bacteria causing hospital nosocomial infection between 2005 and 2007 in our hospital. METHODS:The antibiotic resistance and the data of drug use from 2005 to 2007 in our hospital were analyzed statistically. RESULTS:Over the 3 years,the production rate of ESBLs of Escherichia.coli ascended year by year. The proportion of methicillin resistant staphylococcus aureus (MRSA) in staphylococcus aureus were 79.4%,66.3% and 73.9% respectively over the 3 years. The DDDs of antimicrobials showed an year-on-year increase,with third-generation cephalosporins and fluoroquinolones topping the first two places. CONCLUSION:Antibacterial drugs should be used rationally in clinical practice so as to reduce antibiotic resistance.

BACKGROUND:Mesenchymal stem cells(MSCs) have immunoregulation,can reduce the occurrence rate of graft versus host disease following transplantation,and treat autoimmune disease. OBJECTIVE:To observe the immunoregulation of human umbilical cord(UC) -MSCs on bone marrow T-lymphocyte of patients with aplastic anemia. DESIGN,TIME AND SETTING:A randomized grouping design,in vitro cytological controlled study. Cases were obtained from the Department of Hematology,First Hospital and Second Hospital Affiliated to Nanchang University. Experiments were conducted at the Institute of Hematology,Second Hospital Affiliated to Nanchang University from September 2007 to November 2008. PARTICIPANTS/MATERIALS:A total of 22 patients with aplastic anemia were enrolled at the Department of Hematology,First Hospital and Second Hospital Affiliated to Nanchang University. There were 6 cases of severe aplastic anemia(3 males and 3 females) ,with a median age of 38(16-68) years;16 cases of chronic aplastic anemia(9 males and 7 females) ,with a median age of 41(25-59) years. Umbilical cord was obtained from healthy full-term fetus by normal uterine-incision delivery for isolation,culture and determination of human UC-MSCs. METHODS:MSCs were extracted from human umbilical cord. T-lymphocytes were isolated from bone marrow of patients with aplastic anemia by density gradient centrifugation. MSCs were cocultured with T-lymphocytes. In the control group,T-lymphocytes were incubated at 1?105/well. In the UC-MSCs group,human UC-MSCs were incubated at various densities of 1?105/well,0.5?105/well,0.1?105/well and 0.05?105/well. In the experimental group,T-lymphocytes were cocultured with UC-MSCs at 1:1,1:0.5,1:0.1,1:0.05. MAIN OUTCOME MEASURES:Inhibitory rate of human UC-MSCs at various concentrations on T-lymphocytes of aplastic anemia patients was measured by MTT assay. CD4+/CD8+ changes of the T cells were analyzed by flow cytometry. Expression of the two marrow hematopoietic factors(interleukin-2 and ?-interferon) in the supernatant was evaluated by ELISA. RESULTS:The second passage of MSCs had typical morphology of fibroblast,and highly expressed CD166,CD29,CD54,but lowly expressed CD13,CD34,CD45. MSCs could be induced into adipocytes. UC-MSCs showed inhibitory effect on lymphocyte proliferation of aplastic anemia patients when UC-MSCs and lymphocytes mixed at the ratio of 1:1,0.5:1,0.1:1,and their inhibitory rate were(56.2?12.1) %,(43.7?10.4) %,(28.6?8.9) %. The effect was the strongest at the ratio of 1:1. UC-MSCs adjusted the proportion of CD4+ and CD8+,inhibited the secretion of interleukin-2 and ?-interferon by T cells of aplastic anemia patients. CONCLUSION:Human UC-MSCs can mediate an immunoregulation effect on T lymphocytes of aplastic anemia patients in vitro and the inhibitory effects are dependent on the amount of UC-MSCs.

Objective To establish a method of culturing mesenchymal stem cells from umbilical cord blood. Methods The mononuclear cell fraction of umbilical cord blood was isolated with the aid of a Ficoll/Hypaque gradient, then MSC were cultivated in serum containing medium or serum-free medium. The serum containing medium (SC) consisted of LG-DMEM supplemented with 10% fetal calf serum and 10-6M hydrocortisone, with a final cell concentration of 1?106 cells/ml. The serum-free medium (SF) was supplemented with 1?10 -6M hydrocortisone and 40ng/ml fibronectin with a final cell concentration of 1?106 cells/ml. After a week, 50% of the media were renewed. Before reaching the monolayer phase cells were trypsinised and re-cultured under similar conditions for the another 3 weeks. Cells were counted at the beginning and the end of the culturing period. Phenotypes were identifiad by PAS, NAE, ALP staining. Identification of surface markers and cell cycle analysis of MSCs were performed with flow cytometry. Results Adherent cells appeared 24h after plating of mononuclear cells. The cells formed adherent heterogeneous cell populations after 4-7 days in culture, and they consisted of round and spindle-like cells. In the primary passage of culture, the cells proliferated slowly and became confluent in 14-20 days. When subcultured, the heterogeneous cell populations became homogeneous assuming flat and fibroblast-like shape. Though colonies in the medium containing serum formed earlier than those in the serum-free medium, passage times and cell morphology were the same. The total cell numbers in SF group were lower than those of SC group. The percentage of cells at G0-G1 cell cycle was higher in SF group than that of SC group with significant difference. By flow cytometry analysis, cells of two groups were negative for CD34, CD13 and CD45, but strongly positive for specific surface markers such as CD166, CD29, CD105 and CD54. Conclusion Serum-free medium was superior to medium containing serum for culturing MSCs.

OBJECTIVE:To evaluate the therapeutic effect of clarithromycin on prostatitis induced by NG,CT and U?U.METHODS:56cases of venereal disease-related prostatitis were treated with oral clarithromycin in combination with prostatic massage and hip bath.7days after withdrawing drug,examination of prostatic secretion(EPS)and detection of pathogens were carried out.RESULTS:Total effective rate was82.1%;negative turn rate was92.9%;87.5%EPS routine came back to normal;no obvious ARDs were found.CONCLUSION:Clarithromycin is high in therapeutic effect,slight in ad?verse reactions and convenient in administration.It is suitable for treatment of venereal disease-related prostatitis.

OBJECTIVE: To investigate the association between-1304T/G polymorphism in the promoter of MKK4 gene and the susceptibility in sporadic nasopharyngeal carcinoma. METHOD: MKK4-1304T/G genotypes were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in 90 NPC cases and 30 healthy controls. RESULT: The number of nasopharyngeal carcinoma patients carrying with TG+GG genotype was much higher than those of controls (82.2% vs 66.7%, χ² =10.076, P < 0.05). Analysis showed that compared with the-1304TT genotype, -1304TG heterozygous reduced risk of nasopharyngeal carcinoma 0.56 fold (95% CI = 0.164-1.178, P < 0.01) and-1304GG lower 0.58 fold (95% CI = 0.126-1.381, P < 0.01), TG+ GG genotype variation risk of nasopharyngeal carcinoma decreased 0.72 fold (95% CI = 0.105-0.753, P < 0.01). CONCLUSION MKK4 gene-1304TG genotype can reduce risk of nasopharyngeal carcinoma, and it may be an independent protection factor in sporadic nasopharyngeal carcinoma.
Carcinoma ; Genetic Predisposition to Disease ; Genotype ; Heterozygote ; Humans ; MAP Kinase Kinase 4 ; genetics ; Nasopharyngeal Carcinoma ; Nasopharyngeal Neoplasms ; genetics ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Promoter Regions, Genetic

OBJECTIVE:To optimize the formulation of nanostructured lipid carriers loaded with lornoxicam (LN-NLC). METHODS:Emulsification-solvent evaporation method was used to prepare the LN-NLC. Using drug-lipid ratio,dosage of soy lec-ithin,liquid-lipid ratio (proportion of liquid lipid to total lipid) and dosage of emulsifier as factors,the overall normalized value was calculated by particle size,Zeta potential and entrapment efficiency as indexes was used as comprehensive index. Central com-posite design-response surface method was used to optimize the formulation and investigate the appearance and stability of prepared LN-NLC. RESULTS:The optimal formulation were as follows as drug-lipid ratio of 1:50,dosage of soy lecithin of 162.5 mg,liq-uid-lipid ratio of 25% and emulsifier dosage of 958.2 mg. The particle size of prepared LN-NLC was(96.9±3.3)nm,Zeta poten-tial was(-16.1±0.3)mV,entrapment efficiency was(60.1±0.9)%(n=3),which showed relative error of 2.47%,-4.55%,-0.17%with predicted value,respectively. The prepared LN-NLC was spherical. It had no obvious changes in particle size and Ze-ta potential in sealed storage for 30 d in 4 ℃,and the entrapment efficiency only declined 1.2%. CONCLUSIONS:The LN-NLC formulation is successfully optimized,and the LN-NLC has good stability.

Background and purpose:HPV infection is known as the primary cause of cervical cancer worldwide To investigate high risk type human papillomavirus (HPV) prevalence and incidence rates of cervical intraepithelial neoplasia (CIN) and their screen risk factors in women with different methods of screening.Methods:1137 residents, workers and service women aged 15-59 from Shenzhen city were investigated for cervical cancer in an epidemiology screening study.The high risk types of human papillomavirus of liquid-based cytology samples were tested by hybrid capture 2 (HC-Ⅱ) and liquid-based cytology test (LCT) was also performed at the same time. Women for HPV-positive with LCT ≥ atypical squamous cells of undetermined sign (ASCUS) or HPV-negative with LCT ≥ low grade squamous intraepithelial lesion (LSIL) were biopsied in colposcopy and then were examined by pathology. All data was managed by Foxbase. ?2 test and unconditional Logistic regression model were used for data analysis by SPSS 10.0.Results:1137 women were eligible in our research, the overall rates of HPV infection was 14.0%. HPV detection rates in residents, workers and service women were 14.1%,9.2%,18.9% respectively. HPV detection rates in workers group was significantly lower than that of service women and residents (P