Objective:To explore the effects of superparamagnetic iron oxide-short hairpin RNA ( SPIO-ShRNA) dual functional molecular probes of different concentrations on morphology and biological beha -vior of ovarian cancer SKOV3 cells in vitro.Methods:The dual functional molecular probes at an iron concentration of 5, 15, 30, 45, 75, and 100 mg/L were transfected into SKOV3 cells.The transfection rate of the probe was observed by fluorescence microscope .The distribution and content of iron particles in SKOV3 cells were determined by Prussian blue staining , atomic adsorption spectrometer and electron microscopy .Cell viability was observed by cell counting kit-8 ( CCK-8 ) .The apoptosis was detected by flow cytometry .The expression of protein within the cells was detected by Western blot .The changes of the signal intensity were measured by magnetic resonance imaging (MRI).Results: The SPIO-ShRNA dual functional molecular probe was uptaken in aconcentration-dependence manner within a certain range (5-30 mg/L) .When the concentration of the probe was 45 mg/L, the labeling rate of the cell was close to 100%;With the increase of the concentration of probe , the cell survival rate decreased gradual-ly.The cell survival rate of each experimental group were 94.626%±1.050%, 93.373%±1.180%, 91.700%±3.122%, 75.100%±4.362%, 72.983%±3.233%, 71.010%±2.910%,5, 15, 30mg/L cell survival rate was not significantly decreased , the difference was not statistically significant (P=0.226, P=0.068, P=0.475);When the concentration of the probe was greater than or equal to 45 mg/L,the survival rate decreased obviously ( P<0.001);Group of 45 mg/L protein expression rate was 68.905%± 3.510%, When the concentration of the probe was greater than or equal to 45 mg/L, the inhibition rate of the protein expression level of epidermal growth factor receptor was obviously higher than those of 5, 15, and 30 mg/L groups, the difference was statistically significant (P<0.001, P=0.001, P=0.003, all P<0.01);the MRI displayed that the signal intensity was decreased with increasing concentrations of the probe.The signal intensity of 45 mg/L group was 165.55 ±4.92, compared with the blank control group (same volume of phosphate buffer saline ), normal group(unlabeled ovarian cancer SKOV3 cells), 5, 15, and 30 mg/L groups , the signal intensity of 45 mg/L group decreased significantly (all P<0.001).Con-clusion:The dual functional molecular probe can effectively transfect and specifically inhibit the expression of SKOV3 cell lines at the iron concentration of 45 mg/L, and can also be detected by MRI .The role of diagnosis and treatment of the dual functional molecular probe has been initially confirmed .

Objective To compare the incidence of adverse reaction,clinical manifestation and serious degree of adverse reaction and the intravenous injection time of the two different drugs,and provide references for the safe dosage of the drugs in clinic.Methods 200 patients from June 2009 to June 2010,who visited department of dermatology because of allergic dermatitis were chosen.They were randomly divided into the sodium thiosulfate group and the calcium gluconate group according to the drugs which were injected intravenouly.And incidence of adverse reaction,clinical manifestation and serious degree of adverse reaction and the intravenous injection time were compared between two groups.Results Compared with the calcium gluconate group,the rate of the adverse reaction of the sodium thiosulfate group is lower,and the average time needed is shorter,in addition,there is no serious adverse reaction during injection.Conclusions Intravenous injection of sodium thiosulfate has the advantage of lower incidence of adverse reaction,shorter time needed,and fewer serious adverse reaction,which is worthy of wide spread.

In this study, we investigated the pharmacokinetics parameters of SPIO-shRNA dual functional molecular probe and observed the main organ distribution by MRI in vivo. Eighteen New Zealand white rabbits were randomly divided into three groups and injected intravenously with different doses of SPIO-shRNA molecular probe, respectively. The blood samples were collected to analyze the pharmacokinetic parameters by measuring the iron content at 30 minutes before and after the injection. Twenty-four Kun Ming (KM) mice were randomly divided into 4 groups: the control group was injected intravenously with physiological saline 200 µL per mouse via the tail vein, the other 3 groups were injected intravenously with different doses of SPIO-shRNA molecular probe. MRI observation was performed in 24 hours, and the liver, spleen, kidney, brain and muscle were collected for iron quantification with Prussian blue staining to determine distribution of the SPIO-shRNA molecular probe in the main organ in vivo. Our results suggest that the molecular probe blood half-life is more than 3 hours. The data of MRI suggest the probe was distributed in liver and spleen, and the MRI signal was reduced with the increase in probe's doses (P < 0.05). The results of Prussian blue staining confirmed the results of MRI. Most of the probe could escape the phagocytosis of mononuclear phagocyte system. Our data provide the pharmacokinetic and distribution of SPIO-shRNA molecular probe in organs. Meanwhile, it suggests the choice of the time and dose of probe for MR imaging of tumor in vivo.

AIM: To investigate multi-potential of rat bo ne marrow mesenchymal stem cells (rBMMSC) and mutation inclination, the rBMMSC w ere long passaged in vitro. METHODS: Cellular cycles of diff erent passages were assayed by FA CSan flow cytometry and karyotypes of passage 6, passage 25 and passage 45 were compared by G-binding analysis. RESULTS: The early passages and long-term passages all showed st rong proliferation; passage 6, passage 25 and passage 45 all showed normal karyo type. CONCLUSION: Long-term culture and passage of rBMMSC still remain s strong proliferation. With this capability, the mutation inclination is not enhanced.

Objective Through the analysis of the pathogens distribution of out-patient department and emergency department patients with diarrhea from Shenzhen Hospital of Peking University in 2013~2014,so as to provide reference for the clinical diagnosis and treatment of diarrhea disease.While understanding the improved Salmonella detection results.Methods Var-ied pathogenic bacteria were isolated and identified from 1 719 diarhea stool samples of native Shenzhen Hospital of Peking University from 2013 to 2014 through enrich culture,separate culture,biochemistry,serology etc.Pathogenic virus were test-ed for 451 watery stool specimens by fluorescence PCR.Analyzed statistical differences between the direct inoculation and selenite cystine broth enrichment for Salmonella.Results Picked out 143 disease germs from 1 719 examples diarrhea pa-tient’s stool samples,among which there were 25 strains of ETEC,12 strains of EPEC,8 strains of EAEC,1 strains of EIEC,19 strains ofVibrio parahemolyticus,76 strains Salmonella,2 strains Shigella and 0 strain ofVibrio cholera.There were 10 samples with two disease germs timely.Picked out 189 disease viruses from 451 examples diarrhea patient’s stool without disease germs,among which there were 79 Rotaviruspositive,91 Norwalkvirus positive,9 Adenoviruspositive,10 Astrovirus positive.There were 4 samples with Rotavirus and Norwalkvirus timely.After Salmonella ways to improve the positive rate of 0.6% (17/2 627)increased to 4.4% (76/1 719),χ2=67.2,P<0.01,the difference was statistically signifi-cant.Conclusion The detectable rate of Salmonella and Norwalk virus was the majority,and the clinic enhance the test of the diarrhea pathogenic microorganism,including the improvement of detection method,to reduce the missing rate of them,to provide the scientific basis for the diagnostical therapectic measures.

Objective To develop a sensitive and simple LC/MS method for determination of methomyl in blood samples.Methods Methomyl was extracted from blood by liquid-liquid extraction with ethyl acetate, and then analyzed on a Zorbax SB-C18 (2.1mm×50mm, 5μm) column. The mobile phase consisted of acetonitrile-0.1% formic acid with gradient elution, at a lfow rate of 0.5 mL/min, at 40℃. LC-MS was performed in ESI source with MRM mode for quantiifcation.ResultsThe linear range of the concentration were 0.05~2.0μg/mL for methomyl (r>0.995). The relative recoveries of methomyl were in the range of 90%~108%. The RSDs of intra-days and inter-day were both less than 15%.Conclusion The method is a simple, quick, sensitive and could be used for determination of methomyl in blood samples.

Objective To investigate the duodenum absorptive character of monosialotetrahexosylganglioside sodium (GM1) in rats.Methods The contents of phenolsulfonphthalein (as indicators) and GM1 were determined with ultraviolet-visible (UV) method and high performance liquid chromatography (HPLC) method in rats with in situ cycle intestinal perfusion model.Results The ratio of duodenum absorption of GM1 was 10％ in 2 h after cycle and 22％ in 6 h after cycle,respectively.The Ka was (0.030± 0.012)h,and absorption t1/2 was (25.50 ± 8.56)h in 8 h after cycle.Conclusions GM1 is absorption in rat duodenum,and the accumulate absorption of GM1 is almost linearly related to the cycle time.The absorption dynamics of GM1 may be first-order kinetic process.

Objective: To investigate the characteristics and prognostic factor of central nervous system (CNS) involvement in patients with hemophagocytic lymphohistiocytosis (HLH) . Methods: From January 2006 to October 2015, 152 patients with HLH, 88 patients had CNS involvement, their clinical data were collected, and survival was analyzed using the Kaplan-Meier life table method, univariate and multivariate Cox regression model analyses were applied to identify the risk factors of prognosis. Results: ①57.9% patients complicated with neurological symptoms, cerebrospinal fluid abnormalities were observed in 37.0% patients, 57.5% patients had abnormal neuroradiology. ②36 patients survived well, 3 patients lost to follow-up, 49 dead, 1 survival patient had epilepsy. ③The 3-year overall survival rate of 88 patients was 44%. ④abnormal CSF and unreceived IT bore a significant, independent adverse prognostic value (P<0.05) . Conclusion CNS involvement in HLH has a high frequency and poor prognosis, few patients remained neurologic sequelae; abnormal CSF related to poor prognosis, positive intrathecal injections could improve the prognosis.

Objective:To explore the influencing factors of disability acceptance in patients with spinal cord injury, and analyze the correlation between alexithymia and disability acceptance.Methods:The convenience sampling method was used to select 276 patients with spinal cord injury admitted to Affiliated Hospital of Jining Medical College from October 2018 to June 2020 as the research objects. The general information questionnaire, the Toronto Alexitymia Scale (TAS-20), and the Acceptance of Disability Scale (ADS) was used to conduct the survey, Pearson correlation was used to analyze the correlation between alexitymia and disability acceptance in patients with spinal cord injury, hierarchical regression analysis was used to analyze the factors that affect the disability acceptance of patients with spinal cord injury.Results:The total score of ADS of 276 patients with spinal cord injury was (172.68 ± 37.72) points, and the total score of TAS-20 was (55.60 ± 9.49) points. The results of Pearson correlation analysis showed that all dimensions and total score of TAS-20 were negatively correlated with all dimensions and total score of ADS ( r values were -0.821--0.101, all P<0.05). The results of hierarchical regression analysis showed that gender, course of disease, post-injury working status were the risk factors of disability acceptance ( P<0.05), and alexithymia was the protective factor of disability acceptance( P<0.05), gender, course of disease, post-injury working status and alexithymia level could jointly explain the 22.1% variation in disability acceptance in patients with spinal cord injury. Conclusions:The disability acceptance of patients with spinal cord injury is at a moderate level, and alexitymia is negatively correlated with disability acceptance. alexitymia, gender, course of disease, and post-injury work status are the influencing factors of disability acceptance of patients with spinal cord injury.

Objective: To evaluate the role of tuberculin skin test prified protein derivative (PPD) in defining the screening scope of close contacts of tuberculosis cases in disposal of tuberculosis outbreak in schools. Methods: In a senior middle school in August 2019, 1 553 students of the grade two were tested by PPD because of a school tuberculosis outbreak. PPD results were compared to grade one students without any association with this tuberculosis epidemic, who were also tested by PPD when beginning school. Potential association between PPD distribution characteristics and tuberculosis case distribution were analyzed. Results: Twenty nine grade two students were diagnozed as tuberculosis infection, seven of which were PPD positive, and with the same MIRU-VNTR genotype. In grade one, 0.1 % (1/796) student showed strong PPD positive, 34.3% (273/796) student showed positive. For grade two students, significant higher rate of strong PPD positive [5.9% (45/757)], and PPD positive [52.0% (394/757)] were observed ( χ 2=45.71, 49.90, P <0.01). Proportion of strong PPD positive in the first floor of the teaching building ( 19.4 %), where tuberculosis cases clustered, was significantly higher than that in other floors ( χ 2=89.81, P <0.01); Number of strong PPD positive students increased with TB cases in each floor of the teaching building ( r =0.99, P <0.01). Proportion of strong PPD positive and PPD positive in floors of the dormitory, where TB cases lived, was significantly higher than in other floors ( χ 2=49.4, 64.9 , P <0.01). Number of strong PPD positive and PPD positive students increased with TB cases in each floor of the teaching building ( r =0.84, 0.56, P <0.01). Conclusion Strong PPD positive rate well reflects tuberculosis infection of close contacts, and is recommended for defining the scope of screening.