BACKGROUND:Superparamagnetic iron oxide (SPIO) labeling can trace the migration of stem cells in vivo, and the fluorescent DiI dye is suitable for marking and tracing cells because of its less influence on cellviability, proliferation and differentiation.
OBJECTIVE:To observe the effect and safety of SPIO and fluorescent DiI dye to double label bone marrow mesenchymal stem cells.
METHODS:The bilateral lower limbs of rats were isolated sterilely. Bone marrow was obtained by rinsing using low-glucose DMEM. Bone marrow mesenchymal stem cells were isolated by the whole bone marrow adherence method and purified by differential attachment method. Purified cells were dual-labeled with SPIO particle and fluorescent DiI dye.
RESULTS AND CONCLUSION:Bone marrow mesenchymal stem cells could be separated at 8-10 days after primary culture and the subculturing cycle was 3-4 days. Rat bone marrow mesenchymal stem cells could be effectively labeled with SPIO-DiI and the labeling efficiency was almost 100%. Blue irons contained in intracytoplasmatic vesicles could be observed clearly with Prussian blue staining, and the fluorescence microscopy showed red fluorescence at cytoplasm. Survival and apoptosis percentages obtained by MTT analysis were similar among labeled and unlabeled bone marrow mesenchymal stem cells that were both about 95%.These findings indicate that the rat bone marrow mesenchymal stem cells could be efficiently labeled with SPIO-DiI to construct a nano-bioprobe, without significant changes in morphology, viability and proliferation.

Objective To explore the clinical and serological features of patients with pneumomdiastinum (PNM) and dermatomyositis associated interstitial lung disease (DM-ILD).Methods A total of 145 polymyositis/dermatomyositis (PM/DM) patients hospitalized in our department from March,2010 to December,2012 were recruited.The sera,clinical and laboratory data were collected.Anti-aminoacyl-tRNA synthetase (anti-ARS) and anti-melanoma differentiation-associated gene 5 (MDA5) antibodies were detected by RNA-immunoprecipitation (RNA-IP) and ELISA,respectively.Anti-NXP2,anti-TIF1γ,anti-SRP,anti-SAE and antiMi2 antibodies were detected by immunoprecipitation-Western blotting.Variables were compared between DM patients with and without PNM,as well as between DM-ILD patients with and without PNM.Chi square test,Fisher's exact test,t test,Mann-Whitney U test and a multivariate logistic regression model were used for statistical analysis.Results A total of 11 DM patients (10 patients positive for anti-MDA5 and 1 patient positive for anti-Mi2) developed spontaneous PNM.No PM patient developed PNM.No differences of sex,age at the onset of DM,serum ferritin levels and C reactive protein (CRP) levels was observed between DM patients with and without PNM.Compared with DM patients without PNM,DM patients with PNM had significantly higher frequencies of rapidly progressive ILD (RP-ILD) (63.6％ vs 24.4％,x2=7.25,P=0.01),anti-MDA5 antibodies (90.9％ vs 52.4％,x2=5.86,P=0.02),clinically amyopathic DM (CADM) (63.6％ vs 22.0％,x2=8.57,P=0.007) and cutaneous ulcers (36.4％ vs 11％,x2=5.20,P=0.04),but significantly lower creatine kinase (CK) levels [58.5 (30.5,394.3) U/L vs 284 (73.0,917.0) U/L,t=207.5,P=0.04].The logistic multivariate analysis indicated that cutaneous ulcer was the only independent risk factor for the occurrence of PNM in DM [OR=5.98,95％ confidence interval (CI) (1.12,31.98),P=0.037].In the comparisons between DM-ILD patients with and without PNM,a higher frequency of CADM was observed in the PNM group (63.6％ vs 27.9％,x2=5.37,P=0.03),but no significant difference of the frequencies of RP-ILD,anti-MDA5 antibodies,cutaneous ulcers and CK levels was found in these two groups.All patients with PNM were treated with corticosteroids and immunosuppressants.During the follow-up period,6 patients died of respiratory failure.Conclusion Spontaneous PNM is a refractory complication with poor prognosis,and tends to occur in DM patients with RP-ILD,anti-MDA5 antibody,CADM diagnosis and low CK level,especially in patients with cutaneous ulcers.

Objective To control residual DNA by optimizing methodology during the production of rabies vaccine using Vero cells as a vector .Methods The antigen recovery rate was assessed by linked immunosorbent assay-sandwich technique while the residual DNA was detected by DNA probe hybridization method .Antigen recovery and removal of DNA were the main indexes for evaluateing ultrafiltration , the vital part of rabies vaccine production .Three key factors in ultrafiltration were assessed: selection of membrane packages , ultrafiltration pressure and the concentration ratio .Then protamine was used to pretreat ultrafiltrates .Based on the two indicators mentioned above , the effect of protamine pretreat-ment on the ultrafiltrate was evaluated .Results and Conclusion The optimum condition of ultrafiltration was obtained on the basis of the general antigen recovery rate , DNA removal rate and actual production .The primary parameters of ultrafil-tration were as follows:7.5 ×105 ultrafiltration membrane packages, 20 times concentrated, 15 psi ultrafiltration pressure. After pretreatment with protamine , ultrafiltration has proved to be a molecular sieve in intercepting DNA ,while protamine can tangle the fragmented DNA and form a larger molecular segment , which is believed to be more conducive to ultrafiltra-tion interception .

Objective To investigate the levels of 8 minerals that are related to human health in serum of elderly popu?lation in Tianjin and to provide evidence based guidance for scientifical and rational dietary. Methods Venous blood was collected from 714 elderly people and the levels of 8 minerals were examined by inductively coupled plasma-mass spec?trometry (ICP-MS). Results The percentage of the elderly people who are short of at least one mineral was up to 92.7%. The percentages of deficiency in serum zinc, manganese, magnesium, chrome, iron, calcium, copper, selenium are 79.3%, 73.4%, 58.5%, 42.4%, 39.8%, 23.0%, 1.5%and 0.6%respectively . The level of Ferrum in men’s serum is higher than that in women, while the levels of calcium, chrome and copper in men’s serum are lower than those in women. Conclusion The level of the minerals is generally low in the elderly population from Tianjin and ways were needed in improving dietary.

Objective To investigate the clinical features of polymyositis/dermatomyositis (PM/DM) with anti-nuclear matrix protein (anti-2NXP2) antibodies in Han-Chinese.Methods ImmunoprecipitationWestern Blotting (IP-WB) method was used for screening anti-NXP2 antibodies in 141 adult Han-Chinese patients with PM/DM.The clinical and laboratory data were collected,analyzed and compared with the antiNXP2-positive patients reported in the literature.Statistical analyses were performed using chi-square test and Fisher's exact test.Results Seven (5％) patients,including 6 with DM and 1 with PM,were identified as anti-NXP2 positive.Consistent with the 114 anti-NXP2-positive patients reported in the literature,Han patients with anti-NXP2 had higher frequencies of weakness,heliotrope rash and Gottron's sign.Compared with anti-NXP2-negative patients,anti-NXP2-positive patients presented significantly higher frequencies of dysphagia [43％(3/7) vs 9％(12/134),x2=8.04,P=0.027] and edema [43％(3/7) vs 2％(3/134),x2=26.94,P=0.001 4],while an absence of PM/DM related cancer was observed in Han patients with anti-NXP2.Conclusion Adult Han-Chinese patients with anti-NXP2 are DM predominant,and are characterized by a high frequencies of edema and dysphagia.

Objective To investigate the effect on proliferation and apoptosis by RNA interference to inhibit Nucleostemin(NS) gene expression in gastric carcinoma SGC-7901 cells.Methods The NS-siRNA expression vector was transfected into gastric-carcinoma cells with LipofectamineTM2000 reagent.Then we detected the cell proliferation inhibition ratio by MTT assay,the levels of NS gene expression in all groups by RT-PCR,cell cycle by flow cytometry,cell apoptosis ratio by Annexin-V-FITC/PI kit.Results Compared with that in the control group,cell proliferation in S group decreased;at 24,48 and 72 h the cell proliferation inhibition ratio was 53.21%,71.54% and 87.47%,respectively,the level of NS gene expression reduced in S group.G0/G1 phase cell was 58.34%,S phase cell was 20.68%,and the cell apoptosis was 26.85%.Conclusion RNA interference could substantially inhibit NS gene expression in gastric carcinoma SGC-7901 cells,decrease cell proliferation,arrest cell cycle and increase apoptosis ratio.

Based on a discussion of the major characteristics concerning the flipped classroom model, an in-depth analysis on the four structural elements in the flipped class teaching activities has been made. The teachers are at the same time instructors and teaching resources developers as well, with the feature of team-organizing. As for the students, they become highly-motivated and active learners. The teaching con-tents are profoundly enriched and the teaching media serves as both tools for teaching and learning, and appliance for recognition and communication. It is of great theoretical and instructional significance for us to deeply analyze the structural elements of flipped classroom teaching thus we can contribute not only to the teaching model innovation but also to the current educational reform. Finally, under the continuous de-velopment of emerging technologies, flipped classrooms will surely attract more students to flip learning, which will greatly promote students' learning ability.

Objective:To investigate the promoting effect of Ginsenoside Ro on the differentiation of THP-1-derived dendritic cells (DCs) induced by GM-CSF and IL-4.Methods: Sensitive leukemia-derived DC cell line was screened first.Then,the selected sensitive cell line THP-1 was stimulated to differentiate into DCs by cytokines (GM-CSF and IL-4) and small(5 μmol/L),middle(10 μmol/L),and large (20 μmol/L) dose of Ginsenoside Ro respectively.The expressions of CD1a,MHCⅡ and CD86 of leukemia-derived DCs were detected by flow cytometry.In addition,the transcription levels of CD1a,CD86 and MHCⅡ of leukemia-derived DCs were detected by RT-PCR.ELISA was used to measure the protein levels of TNF-α and IL-6 in the culture supernatant.Results: THP-1 was the sensitive leukemia cell line which could be induced to differentiate into DCs by cytokines.Compared with cytokine stimulation alone,the expression of CD1a,MHCⅡ and CD86 in leukemia-derived cells was significantly increased after the stimulation of Ginsenoside Ro combined with cytokine(P<0.05).The CD1a,CD86 and MHCⅡ mRNA expression was significantly increased after the treatment of Ginsenoside Ro combined with cytokine(P<0.05).Moreover,the protein levels of TNF-α and IL-6 in culture supernatant were significantly increased (P<0.05) after the stimulation of Ginsenoside Ro in combination with cytokines.Conclusion: Ginsenoside Ro can significantly promote the differentiation of leukemia-derived DCs.

Objective To explore the correlation between thyroid nodules and uric acid levels and to find their gender differences.Methods A total of 68 056 subjects in a regional medical physical examination center of Shanxi Province from January 2013 to June 2015 were enrolled in this study.All the participants′ general information and parameters were recorded.Thyroid nodules were detected by color Doppler ultrasonography.Results The total prevalence of thyroid nodule was 35.5%, 30.7% in males and 40.0% in females.The prevalence of single nodule was 50.1%, and multiple 49.9%.Compared with no nodule group, thyroid nodule group tended to be older, with higher BMI, and with a worse metabolic status(all P<0.01).The uric acid levels were lower[(352.37±78.14 vs 357.70±77.51) μmol/L, P<0.01] in thyroid nodule group in male and higher[(260.22±61.91 vs 253.91±59.18) μmol/L, P<0.01] in female.Conclusion Thyroid nodules may be associated with metabolism and inflammation.In males, hyperuricemia group had lower, while in females, hyperuricemia ones were with a higher prevalence of thyroid nodules.

Objective:To evaluate the feasibility and safety of uniportal video-assisted thorascopic pneumonectomy for non-small cell lung cancer patients, as well as the surgical techniques.Methods:From January 2017 to June 2018, 23 non-small cell lung cancer patients who underwent uniportal VATS pneumonectomy in our hospotal were retrospectively evaluated. There were 20 males and 3 females with a mean age of(60.3±6.0) years. The clinical data, complications and operative mortality were analyzed.Results:All 23 patients with lung cancer were operated safely. There was no conversion to thoracotomy. The mean operating time was(160.8±65.0) min. The mean blood loss was(131.3±121.3) ml. The mean hospital stays were(14.0±6.3) days. The mean postoperative hospital stays were(8.0±4.5) days. Postoperative morbidity occurred in 4 patients(17.4%). There was no perioperative mortality. Pathology examination of all patients were lung cancer(5 cases of adenocarcinoma, 15 cases of squamous carcinoma, 2 cases of large cell carcinom, 1 case of adenocarcinoma combined with large cell carcinoma).Conclusion:Uniportal video-assisted thorascopic pneumonectomy is a feasible operation that could be performed safely in a selected group of patients by experienced surgeons.