BACKGROUND: Reports of culture condition of olfactory ensheathing cells (OECs) vary. And some methods have bad reproducibility, not appropriate for actual application.OBJECTIVE: To observe the effect of differential attachment, chemicals in combination with trypsin digestion to purify rat OECs in vitro and to compare the effect with differential attachment or chemicals alone.DESIGN, TIME AND SETTING: In vitro controlled observation of cytology was performed at the Laboratory of Department of Human Anatomy & Histology and Embryology, Fujian Medical University from June 2007 to June 2008.MATERIALS: A total of 8 SD rats, 2 days old, were provided by the Laboratory Animal Center of Fujian Medical University.METHODS: The OECs were dissociated from the postnatal rat olfactory bulbs under sterile condition, and seeded in poly-L-lysine-coated culture flask at a density of 4.0×10~8 /L for primary culture. The cells were divided into 4 groups: control group (not purified); chemicals group (5 μmol/L arabinose); differential attachement group (Nash differential attachement); combination group (Nash differential attachement to eliminate most of the fibroblasts, followed by arabinose; when the cells were confluent at 6 days, the cells were digested with 1.25 g/L trypsin for 1 minute until the processes were shrank, cells became round, with some cells floating).MAIN OUTCOME MEASURES: Morphological changes of the cultured OECs and NGFRp75 immunocytochemistry were observed.RESULTS: The OECs displayed a very characteristic morphological appearance. Most of OECs were bipolar or tripolar with long and slim processes. In the unpurified control group the rapidly proliferating fibroblasts were in the majority (60%) after 7 days in culture, and confluent at day 14. The OECs occupied the most in the other groups, and their appearance remained unchanged.The surviving bipolar or tripolar OECs were positive for NGFRp75. The purifity by chemicals and differential attacehment methods was low (75%), while the combination group was high (85%).CONCLUSION: The method of purifing OECs through a combination of differential attacehment, chemicals and trypsinization is effective.

Objective To discuss clinical application of theory of TCM constitution on non-alcoholic fatty liver disease (NAFLD).Methods Literature of nearly 15 years was summarized to analyze the development of Chinese constitution theory and its application of on typing and treatment of NAFLD.Results Main types of constitution in patients with NAFLD were phlegm-dampness,deficiency of Qi and damp-heat.On tiis basis,treatment based syndrome differentiation of TCM could effectively prevent and treat NAFLD.Conclusion TCM constitution theory provide research ideas for NAFLD.

Objective To compare the therapeutic effect of intraarticular injection with sodium hyaluronate (SH) and SH combined with triamcinolone acetonide for osteoarthritis (OA) of knee joint.Methods Eighty-two patients with OA of knee joint were divided into two groups by toss a coin from March 2010 to March 2011.Forty-two patients simplely accepted SH injection (SH group),40 patients accepted SH combined with triamcinolone acetonide (combination group).Intraarticular injection was taken once a week,a total of 5 times.Efficacy measurements included visual analog scale (VAS) for pain and safety.Results After treatment for 1 week,the measured value of VAS for up and down stairs in combination group was significantly lower than that in SH group [ (22.6 ± 6.1 ) mm vs.(63.9 ± 5.8 ) mm] (P ＜ 0.05 ).After treatment for 3,6 months,the measured value of VAS for up and down stairs in two groups had no significant difference [ (42.2 ± 6.6) mm vs.(41.1 ± 5.3) mm,(48.5 ± 5.5) mm vs.(49.3 ± 6.2) mm] (P ＞ 0.05).Function of the knee joint after treatment was better than before treatment in two groups.Adverse effects were minimal. Conclusions Intraarticular injection of SH and SH combined with triamcinolone acetonide OA of knee joint is a safe and effective method.SH combined with triamcinolone acetonide can improve the efficacy in early period.

Objective To evaluate the efficiency of glycogen phosphorylase BB (GPBB) and heart-type fatty acid binding protein (H-FABP) in the early diagnosis of neonatal myocardial injury (NMI). Methods The GPBB and H-FABP levels in the blood were detected at 3, 6, 12 and 24 h after onset of the NMI. The GPBB and H-FABP levels were compared among NMI, Non-NMI and control groups. The NMI diagnosis sensitivity and specificity for GPBB, H-FABP, GPBB combined with H-FABP were compared. The relationships of GPBB, H-FABP levels with the risk factors were analyzed. Results The GPBB and H-FABP levels of NMI group were higher than those of Non-NMI and control groups (P < 0.01). The diagnosis sensitivity of four time points (3, 6, 12 and 24 h) of combined GPBB and H-FABP were higher than those of the GPBB and H-FABP (P < 0.05). The GPBB and H-FABP were negatively correlated with blood glucose level, positively correlated with asphyxia degree and septicemia degree. Conclusion The combined application of GPBB and H-FABP can improve sensitivity in the early diagnosis of NMI.

In the item bank construction of medical microbiology examination questions,the accurate difficulty coefficient is an important parameter to guarantee the quality of the bank.Thus,the accurate difficulty estimate plays an important role in the item bank construction of examination questions.Methods of estimating the difficulty of medical microbiology test questions were explored,based on the knowledge points,the type of test questions,the structure of test questions and the degree of students’ familiarity with the questions and so on.Finally,the feasibility of methods was confirmed through the simulated tests.

Objective To optimize the formulation of the sunscreen used in military training.Methods The different formulations of sunscreen were prepared according to the orthogonal design,and then applied on 3M adhesive tape posted on the color dish with a certain amount;the absorbance in 280 nm-320 nm and 320 nm-400 nm of them were measured and the arithmetic mean (A) was concluded.By comparing A,the protective effect and the main factors affecting preventive bask were determined.The preventive bask test of the optimum formula was carried out in human trials using the method of multi point increasing ultraviolet irradiation.Results OMC,TinosorbTM M,rutin,pearl powder and baicalin all had significant effects in preventing UVA.OMC had a significant effect in preventing UVB.The formula 1 (containing the five materials mentioned above) had the best effects in preventing both UVA (SPF 18) and UVB (PA+).Conclusions The formula 1 has the best effects in preventing both UVA and UVB.

Aim To investigate the effects of aloe coarse polysaccharid on proliferation and the intracellular calcium level and the leakage rate of lactate dehydrogen-ase(LDH) of the epidermal cellsin vitro.Methods The epidermal cells were treated with aloe coarse po-lysaccharid of75,150,300,600 and 1 200 mg.L-1respectively.The proliferation of epidermal cells wasobserved interms of the proliferation curve and the pop-ulation doubling/day(PD/d).The concentration ofintracellular calcium and the leakage rate of LDHwerealso measured.Results In aloe coarse polysaccharidgroup,a dose-dependent and statistically significant(P

The previous pharmacokinetic methods can be only limited to drug analysis in vitro, which provide less information on the distribution and metabolismof drugs, and limit the interpretation and assessment of pharmacokinetics, the determination of metabolic principles, and evaluation of treatment effect. The objective of the study was to investigate the pharmacokinetic characteristics of gene recombination angiogenesis inhibitor Kringle 5 in vivo. The SPECT/CT and specific 131I-Kringle 5 marked by Iodogen method were both applied to explore the pharmacokinetic characteristics of 131I-Kringle 5 in vivo, and to investigate the dynamic distributions of 131I-Kringle 5 in target organs. Labeling recombinant angio-genesis inhibitor Kringle 5 using 131I with longer half-life and imaging in vivo using SPECT instead of PET, could overcome the limitations of previous methods. When the doses of 131I-Kringle 5 were 10.0, 7.5 and 5.0 g/kg, respectively, the two-compartment open models can be determined within all the metabolic process in vivo. There were no significant differences in t1/2α, t1/2β, apparent volume of distribution and CL between those three levels. The ratio of AUC(0 ? 1) among three different groups of 10.0, 7.5 and 5.0 g/kg was 2.56:1.44:1.0, which was close to the ratio (2:1.5:1.0). It could be clear that in the range of 5.0–10.0 g/kg, Kringle 5 was characterized by the first-order pharmacokinetics. Approximately 30 min after 131I-Kringle 5 was injected, 131I-Kringle 5 could be observed to concentrate in the heart, kidneys, liver and other organs by means of planar imaging and tomography. After 1 h of being injected, more radionuclide retained in the bladder, but not in intestinal. It could be concluded that 131I-Kringle 5 is mainly excreted through the kidneys. About 2 h after the injection of 131I-Kringle 5, the radionuclide in the heart, kidneys, liver and other organs was gradually reduced, while more radionuclide was concentrated in the bladder. The radionuclide was completely metabolized within 24 h, and the distribution of radioactivity in rats was similar to normal levels. In our study, the specific marker 131I-Kringle 5 and SPECT/CT were suc-cessfully used to explore pharmacokinetic characteristics of Kringle 5 in rats. The study could provide a new evaluation platform of the specific, in vivo and real-time functional imaging and pharmacokinetics for the clinical application of 131I-Kringle 5.

Objective:To develop and validate a clinical prediction model for the risk of malignant ventricular arrhythmia in patients with acute myocardial infarction (AMI) during hospitalization, and evaluate the effect of the prediction model.Methods:A retrospective study was conducted. A total of 2 649 patients with AMI admitted to cardiology department of Changzhou No.2 People's Hospital of Nanjing Medical University from December 2012 to August 2020 were enrolled. The clinical characteristics including gender, age, medical history, discharge diagnosis, vital signs during hospitalization, electrocardiogram characteristics at admission, laboratory examination indexes, interventional treatment, drug usage, malignant ventricular arrhythmias [mainly included sustained ventricular tachycardia (VT), ventricular flutter or ventricular fibrillation (VF)], and death were recorded. All patients were divided into two groups according to whether VT/VF occurred during their hospitalization. Independent risk factors for VT/VF during hospitalization were evaluated by multivariate Logistic regression analysis, and a clinical prediction model was constructed. The receiver operating characteristic curve (ROC curve) was plotted, and the area under ROC curve (AUC) was calculated to evaluate the accuracy of the prediction model.Results:A total of 2 649 eligible patients with AMI were enrolled, of whom 134 (5.06%) developed VT/VF during hospitalization. The in-hospital mortality rate in VT/VF group was significantly higher than that in non-VT/VF group (38.1% vs. 1.7%, P < 0.01). Compared with the non-VT/VF group, the patients in the VT/VF group with lower systolic blood pressure [SBP (mmHg, 1 mmHg = 0.133 kPa): 125.9±28.2 vs. 132.0±24.2], higher random blood glucose (mmol/L: 8.6±4.8 vs. 7.4±3.7), worse cardiac function [Killip heart function grade ≥ 3: 36.6% vs. 10.7%, left ventricular ejection fraction (LVEF) < 0.50: 56.7% vs. 33.6%, frequent premature ventricular contractions: 12.7% vs. 1.2%] and more hypokalemia (46.3% vs. 17.3%), with significant differences (all P < 0.05). Multivariate Logistic regression analysis showed that Killip classification of cardiac function ≥ 3 [odds ratio ( OR) = 3.540, 95% confidence interval (95% CI) was 2.336-5.363], random blood glucose > 11.1 mmol/L ( OR = 1.841, 95% CI was 1.171-2.893), LVEF < 0.50 ( OR = 0.546, 95% CI was 0.374-0.797), frequent premature ventricular contractions ( OR = 12.361, 95% CI was 6.077-25.144), potassium < 3.5 mmol/L ( OR = 4.268, 95% CI was 2.910-6.259), SBP < 90 mmHg ( OR = 0.299, 95% CI was 0.150-0.597) and creatinine (Cr) > 100 μmol/L ( OR = 2.498, 95% CI was 1.170-5.334) were independent risk factors for VT/VF in patients with AMI (all P < 0.05). The clinical prediction model of VT/VF risk was constructed based on the variables selected by multivariate regression analysis. The ROC curve analysis showed that the AUC of the model in predicting VT/VF was 0.779 (95% CI was 0.735-0.823, P < 0.001); the optimal cut-off value of the model was 17, the sensitivity was 76.1%, the specificity was 67.3%. Conclusions:The incidence of VT/VF during hospitalization of AMI patients significantly increases the risk of in-hospital death. The independent risk factors of VT/VF are Killip grade ≥ 3, random blood glucose > 11.1 mmol/L, LVEF < 0.50, frequent ventricular premature beats, potassium < 3.5 mmol/L, SBP < 90 mmHg and Cr > 100 μmol/L. The newly constructed clinical prediction model has certain predictive value for the occurrence risk of VT/VF.

Objective:To establish the fingerprints of the best anti-MDR-MRSA components in the combination of honeysuckle and forsythia. Methods:The best active anti-MDR-MRSA ingredients of honeysuckle and forsythia were dissolved in 10% methanol water. A Waters Atlantis T3 (250 mm × 4. 6 mm, 5 μm) column was eluted with water and methanol as the mobile phase. The optimal chro-matographic conditions were determined by optimizing the mobile phase and chromatographic conditions. The fingerprints similarity of 10 batches of active anti-MDR-MRSA components in honeysuckle and forsythia was studied. Results:The best active ingredients of the combination of honeysuckle and forsythia showed 7 peaks with promising separation. The similarity of the 10 samples was greater than 0. 9. Conclusion:The fingerprints of the best active anti-MDR-MRSA ingredient in the combination of honeysuckle and forsythia are stable, and can be used for the quality evaluation of the components.