Abstract: Transfection of Plasmodium falciparum is helpful to study the function of its genes, such as drug resistance. However, transgenic manipulation has been very challenging, mainly due to the high A/T base sequence structure (A+T content of about 82%) and low transfection efficiency of the Plasmodium genome. Electroporation-based transfection of Plasmodium falciparum has been successfully applied in the study of certain genes, and electroporation by preloading is currently the preferred method for introducing foreign DNA into Plasmodium falciparum. The site-directed editing of Plasmodium genes mostly adopts the method of two-plasmid transfection. It is generally believed that successful transfection of Plasmodium requires a large amount of high-purity plasmid DNA and an accurate transfection system. In addition to the evaluation of the current commonly used electrotransfection methods, this paper also introduces a new transfection method, namely lyse-reseal erythrocytes for transfection (LyRET). This paper also review the role of factors such as plasmid DNA concentration, the use of transfection reagents, the setting of transfection parameters, the addition of fresh red blood cells, and the markers of successful transfection in improving the success rate and efficiency of Plasmodium transfection, in the hope of providing a reference for study in this field.

This study was aimed to explore the role of human fetal bone marrow stromal cells (FBMSC) in combination with exogenous cytokines in supporting the in vitro expansion of cord blood mononuclear cells and the expression of CXCR4(+) and CD49d(+) in CD34(+) cells. Mononuclear cells (MNC) separated from cord blood (CB) were cultured in a serum-free support culture system with FBMSC or exogenous cytokines or both of them. On day 0, 6, 10 and 14, total cells were counted, CD34(+), CD34(+)CXCR4(+) and CD34(+)CD49d(+) cells were quantitated by FACS, and hematopoietic progenitor cells were assessed by semisolid culture assay. The results showed that after culturing for 14 days, CD34(+) cells, CD34(+)CXCR4(+) cells, CD34(+) CD49d(+) cells and colony forming unit (CFU) were significantly increased (P < 0.05). Compared with other groups, expansion multiple of CD34(+), CD34(+)CXCR4(+), CD34(+)CD49d(+) cells and CFU were higher than that in FBMSC and cytokine group (P < 0.05). It is concluded that the culture system used in this study can not only support the expansion of CB MNCs but also increase the number of hematopoietic stem and progenitor cells which has chemokine and adhesion capacity. This culture system may be a feasible way for in vitro culture of cord blood cells.
Antigens, CD34 ; blood ; Bone Marrow Cells ; cytology ; immunology ; Cell Proliferation ; drug effects ; Cells, Cultured ; Coculture Techniques ; Cytokines ; pharmacology ; Fetal Blood ; cytology ; Fetus ; Flow Cytometry ; Granulocyte-Macrophage Colony-Stimulating Factor ; pharmacology ; Humans ; Integrin alpha4 ; blood ; Interleukin-3 ; pharmacology ; Leukocytes, Mononuclear ; cytology ; immunology ; Receptors, CXCR4 ; blood ; Stromal Cells ; cytology ; immunology ; Time Factors

BACKGROUNDPrevious researches about necrotic pancreatic tissue infections are numerous, but the study on systemic infection related to the severe acute pancreatitis (SAP) treatment period is limited. This study aimed to investigate the distribution and drug resistance of pathogenic bacteria in patients who had hepatobiliary surgery for SAP during the past three years.

METHODSA retrospective study was conducted on the distribution, category and drug resistance of pathogenic bacteria in patients who had hepatobiliary surgery for SAP from 2008 to 2011.

RESULTSA total of 594 pathogenic bacteria samples were isolated. Among them 418 isolates (70.4%) were Gram bacteria negative, 142 isolates (23.9%) were Gram bacteria positive, and 34 isolates (5.7%) were found fungi. The most common Gram negative bacteria were Escherichia coli (19.8%), and the dominant Gram positive pathogenic bacteria were Enterococcus faecium. The distribution of SAP-related infectious pathogens was mainly in peritoneal drainage fluid, sputum, bile, and wound secretions. Almost all the Gram negative pathogenic bacteria were sensitive to carbapenum. Extended-spectrum β-lactamases (ESBLs) producing strains were more resistant to penicillins and cephalosprins than the ESBLs non-producing strains. Staphylococcus was sensitive to vancomycin and linezolid. The drug resistance of meticillin-resistant staphylococcus (MRS) to commonly used antibiotics was higher than meticillin-sensitive streptococcus (MSS). Enterococcus sp. exhibited lower drug-resistance rates to vancomycin and linezolid.

CONCLUSIONSGram negative bacteria were the dominant SAP-related infection after hepatobiliary surgery. A high number of fungal infections were reported. Drug resistant rates were high. Rational use of antibiotics according to the site of infection, bacterial species and drug sensitivity, correctly executing the course of treatment and enhancing hand washing will contribute to therapy and prevention of SAP-related infection and decrease its mortality.

Anti-Bacterial Agents ; pharmacology ; Gram-Negative Bacteria ; drug effects ; Gram-Positive Bacteria ; drug effects ; pathogenicity ; Humans ; Microbial Sensitivity Tests ; Pancreatitis ; microbiology

OBJECTIVETo study the prevalence of chondromalacia patella among college students and the correlation with sports injury.

METHODS354 students from gymnastic department and 429 from nongymnastic department with knee joint pain were selected. 184 students from gymnastic department and 342 from nongymnastic department were checked randomly by a surgeon. 77 patients (37 males, 40 females) from gymnastic department and 119 patients (62 males, 57 females) from nongymnastic department were diagnosed as chondromalacia patellae. The amount of exercise and the occurrence of sports injury were investigated in each student. All data were analyzed with SPSS 10.0 statistical software.

RESULTSThe prevalence of chondromalacia patella was 20.1% in female students and 11.6% in male students from gymnastic department, and 5.61% in female students and 4.92% in male students from nongymnastic department. The amount of exercise and the occurrence of sports injury to the knee joint in students from gymnastic department were greater than those from nongymnastic department.

CONCLUSIONSIn both female and male students, the prevalence of chondromalacia patella is higher in gymnastic department than nongymnastic department. Sports injury is an important cause of chondromalacia patella.

Adolescent ; Adult ; Age Distribution ; Athletic Injuries ; epidemiology ; pathology ; Cartilage Diseases ; diagnosis ; epidemiology ; Case-Control Studies ; China ; epidemiology ; Comorbidity ; Cross-Sectional Studies ; Female ; Humans ; Injury Severity Score ; Knee Injuries ; epidemiology ; pathology ; Male ; Patella ; pathology ; Prevalence ; Probability ; Prognosis ; Reference Values ; Risk Factors ; Sex Distribution

Objective To explore the inter-regional,base-like and informatized support of the field medical station during rotational training.Methods The field medical station information system developed by the hospital was introduced,which had the working mode involving in a set of system and two kinds of terminals.The problems of the information system were analyzed during iner-regional,base-like rotational training.Results The information system had its functions realized,and stills had to be improved in casualty information input flow,precision materials management and allocation standard of operating terminal.Conclusion The field medical station information system contributes to enhancing its service efficiency and informatization.

Objective To explore the feasibility of evaluating the lung function by MSCT in emphysema.Methods The MSCT scan and pulmonary function tests(PFF)were respectively performed in 147 receptors within one week.They were randomly divided into 2 groups:group A(120 receptors), including normal,mild,moderate and severe abnormal pulmonary function based on the PFT,for comparing the correlation between pulmonary quantitative indexes of MSCT pulmonary function and PFT and settingup the primary grade criteria of abnormal pulmonary function in emphysema,group B(27 receptors)for evaluating the diagnostic accuracy in group A.The total lung was respectively scanned at the full inspiration and full expiration with MSCT.The pulmonary quantitative indexes of MSCT were measured with Siemens Pulmo pulmonary quantitative software.Results There was correlation between pulmonary quantitative indexes of MSCT and PFF.The Piex/in_(-910)showed best correlation with FEV_1%(r=-0.905,P

OBJECTIVETo determine the efficacy of the plant-derived bioflavonoid, quercetin, for treating nonalcoholic fatty liver disease (NAFLD) by using a rat model, and to investigate the molecular mechanism underlying its therapeutic effects.

METHODSOne-hundred Sprague-Dawley rats were randomly assigned into the normal control group (normal group), untreated NAFLD model control group (model group), 75 mg/kg/day quercetin treatment group (low-dose group), and 300 mg/kg/day quercetin treatment group (high-dose group). The NAFLD rat model was established by providing four weeks of a high-fat diet; the normal group received normal rat chow diet. The quercetin treatments were administered for eight weeks after model establishment and control groups received simultaneous gavages of isotonic saline, with continuation of the respective diets. At the end of the eight weeks (experimental week 12), the rats were sacrificed for liver and serum collection. Intergroup differences in liver index, fasting blood glucose (FBG), triglycerides (TG), interleukin (IL)-18, IL-10, malondialdehyde (MDA), and histopathological features were assessed by independent samples t-test (normal vs. model), one-way ANOVA (model vs. treatments), and least significant difference t-test (pairwise comparisons); correlations were assessed by Pearson's correlation coefficient.

RESULTSCompared with the normal group, the model group showed significantly higher liver index (t=-2.327), FBG (t=-3.482), TG (t=-0.302), and serum IL-18 (t=-2.704) (all P less than 0.05), but significantly lower IL-10 (t=2.622, P less than 0.05); the MDA level was also higher in the model group, but the difference was not significant (t=-1.083, P less than 0.05). Livers from the model group showed obvious histological features of inflammation (lymphocyte and neutrophil infiltration) and steatosis (cytoplasmic lipid droplets). Inflammation was positively correlated with IL-18 (P less than 0.05), but negatively correlated with IL-10 (P less than 0.05), while steatosis was negatively correlated with IL-10 (P less than 0.05). Compared to the model group, quercetin treatment (both low- and high-dose) led to significant decreases in the liver index, FBG and IL-18 (all, P less than 0.01), and significant increase in IL-10 (P less than 0.05); however, the changes in liver index, FBG and IL-10 were not significantly different between the low- and high-dose treatment groups, but the high-dose of quercetin did induce a significantly greater decrease in IL-18 than the low-dose (P less than 0.05).

CONCLUSIONNAFLD rats have higher serum levels of IL-18 but lower levels of IL-10 than their healthy counterparts, and these differential cytokine expressions may be related to liver inflammation and steatosis. Quercetin treatment may help to delay the progression of NAFLD, possibly by adjusting the balance of inflammatory cytokines.

Animals ; Fatty Liver ; blood ; drug therapy ; Interleukin-10 ; blood ; Interleukin-18 ; blood ; Male ; Non-alcoholic Fatty Liver Disease ; Quercetin ; pharmacology ; therapeutic use ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the effects of quercetin on serum levels of resistin and interleukin (IL)-18 and incidence of insulin resistance (IR) in nonalcoholic fatty liver disease (NAFLD) using a rat model.

METHODSNAFLD was induced in Sprague-Dawley rats by administering a high-fat diet for four weeks. The model rats were then treated with quercetin (oral gavage administration; low dose group: 75 mg/kg/day, high dose group: 300 mg/kg/day) for eight weeks. Untreated model rats served as controls. Serum levels of resistin, triglyceride (TG), IL-18, fasting plasma glucose (FPG), fasting insulin (FINS), and malondialdehyde (MDA) were measured by standard biochemical assays before and after the quercetin administration. In addition, the insulin resistance index (HOMA-IR) was calculated and pathological changes in liver were observed by histological analysis.

RESULTSCompared to the untreated model rats, the quercetin treated model rats showed significantly lower serum resistin (5.98 vs. 2.70), serum IL-18 (10.93 vs. 8.21), FPG (7.45 vs. 4.99), FINS (12.69 vs. 8.59), and HOMA-IR (4.22 vs. 1.87) (all P less than 0.01). Compared to the untreated model group, the high dose group showed significantly lower TG (t = 4.70) and MDA (t = 5.14) (both P less than 0.01). Serum levels of resistin and IL-18, and levels of TG, FPG and FINS were found to be positively correlated with HOMA-IR and the degree of liver disease (r more than 0, all P less than 0.05). The degree of degeneration was decreased in accordance with the dosages of quercetin, as compared to the untreated model group (U = 4.41 and 2.19, both P less than 0.05), and the pathological degree was less extensive in the high dose group than in the low dose group (U = 2.44, P less than 0.01).

CONCLUSIONQuercetin treatment reduces levels of inflammatory cytokines and improves lipid peroxidation and IR in NAFLD rats, and its beneficial effects appear to increase with higher dosage.

Animals ; Insulin Resistance ; Interleukin-18 ; blood ; Non-alcoholic Fatty Liver Disease ; Quercetin ; Rats ; Rats, Sprague-Dawley ; Resistin

With the wide application of electronic data management (EDC), the data management is shifting to a new mode. In order to recognize the advantages of EDC, we choose 20 representative registered clinical trials, which involve 5 404 subjects and 321 sites. We found that EDC has many beneficial impacts on the course of clinical trial data management, including the process of data collection, data cleaning, data quality control and clinical trial decision-making. The result also provides a reference for the adoption of EDC in clinical trials.
Clinical Trials as Topic ; Data Collection ; standards ; Information Storage and Retrieval ; standards ; Quality Control

OBJECTIVETo assess the reliability and validity of the Aging Males' Symptoms (AMS) scale in the male population of Shanghai.

METHODSWe enrolled 973 males aged 40 years and over in a community of Shanghai, China. Using the AMS scale, we calculated the split-half reliability coefficient and Cronbach's alpha coefficient, assessed the validity through confirmatory factor analysis and correlation analysis, and obtained the domain scores of different people by analysis of variance and independent sample test.

RESULTSThe split-half reliability was > 0.78 (P < 0.01) and Cronbach's alpha coefficients of all the dimensions > 0.82 (P < 0.01). Confirmatory factor analysis showed 3 domains in the AMS scale, Pearson correlation coefficients of all the items to their domains were > 0.49 (P < 0.01), and the total testosterone level was not correlated with AMS scores, with Pearson correlation coefficient of -0.04 (P > 0.05). Statistically significant differences were found in AMS scores among different age groups as well as among those with different chronic disease histories, but not in the psychological domain among different age groups.

CONCLUSIONThe reliability and validity of the AMS scale are acceptable in assessing aging males'symptoms among the male population of Shanghai, but further studies are needed to determine whether it could be used as a tool for screening late-onset hypogonadism (LOH) in males.

Adult ; Aged ; Aging ; China ; epidemiology ; Humans ; Hypogonadism ; epidemiology ; psychology ; Male ; Middle Aged ; Psychometrics ; Reproducibility of Results ; Surveys and Questionnaires