Epidemiological characteristics of public health emergencies of infectious diseases in schools of Beijing from 2010 to 2020 / 中国学校卫生

Objective@#To analyze the epidemiological characteristics of infectious diseases at schools in Beijing from 2010 to 2020, providing evidence for the prevention and control strategies in school infectious diseases.@*Methods@#Information on public health emergencies was collected from Public Health Emergency Reporting System (the subsystem of Chinese Disease Prevention and Control Information System) reported from 2010 to 2020.@*Results@#A total of 146 public health emergencies and 138 public health emergencies of infectious diseases in schools were reported in Beijing from 2010 to 2020, including 4 291 cases with the rate of 2.32% and affected or exposed 185 179 cases. There were significant difference in mean annual incidence rates( χ 2=782.46, P <0.01). There were 71 events of respiratory infectious diseases and 66 events of intestinal infectious diseases, accounting for 51.45% and 47.83%, respectively. The annual incidence peaks of public health emergencies of infectious diseases were during March-June and October-December. The events mainly occurred in kindergartens and primary schools among each stage of school periods with 51 and 46 incidences respectively, which accounted for 70.29% of the total number of public health emergencies in schools. The leading infectious diseases among all the reported events in kindergartens and primary schools were hand foot mouth disease and varicella. Varicella and other infectious diarrhoeal diseases were at the top lists of infectious disease outbreaks at the secondary and university stages.@*Conclusion@#Infectious diseases events were the major type of public health emergencies at schools in Beijing from 2010 to 2020. Respiratory infectious diseases and intestinal infectious diseases were the keys to the prevention and control of public health emergencies related to school. It is necessary to strengthen the surveillance for public health emergencies especially for symptom surveillance. The prevention and control measures should be taken according to the characteristics of different age groups. At the same time, the prevention and control of school infectious diseases and the drill of the plan during peak periods need to be particularly strengthened.

The Value of CD44 / 中国实验血液学杂志

OBJECTIVE@#To investigate the value of CD44@*METHODS@#Flow cytometry was used to detected the proportion of CD44@*RESULTS@#The percentage of CD44@*CONCLUSION@#HCD44

Relationship between Leukocytes Derived Microparticles and Minimal Residual Disease and Prognosis of Acute Myeloid Leukemia / 中国实验血液学杂志

OBJECTIVE@#To detect the relationship between leukocytes derived microparticle (CD45@*METHODS@#The expression of CD45@*RESULTS@#The percentages of CD45@*CONCLUSION@#High level of CD45

Bone marrow-derived mesenchymal stem cells modulate autophagy in RAW264.7 macrophages via the phosphoinositide 3-kinase/protein kinase B/heme oxygenase-1 signaling pathway under oxygen-glucose deprivation/restoration conditions / 中华医学杂志(英文版)

BACKGROUND@#Autophagy of alveolar macrophages is a crucial process in ischemia/reperfusion injury-induced acute lung injury (ALI). Bone marrow-derived mesenchymal stem cells (BM-MSCs) are multipotent cells with the potential for repairing injured sites and regulating autophagy. This study was to investigate the influence of BM-MSCs on autophagy of macrophages in the oxygen-glucose deprivation/restoration (OGD/R) microenvironment and to explore the potential mechanism.@*METHODS@#We established a co-culture system of macrophages (RAW264.7) with BM-MSCs under OGD/R conditions in vitro. RAW264.7 cells were transfected with recombinant adenovirus (Ad-mCherry-GFP-LC3B) and autophagic status of RAW264.7 cells was observed under a fluorescence microscope. Autophagy-related proteins light chain 3 (LC3)-I, LC3-II, and p62 in RAW264.7 cells were detected by Western blotting. We used microarray expression analysis to identify the differently expressed genes between OGD/R treated macrophages and macrophages co-culture with BM-MSCs. We investigated the gene heme oxygenase-1 (HO-1), which is downstream of the phosphoinositide 3-kinase/protein kinase B (PI3K/Akt) signaling pathway.@*RESULTS@#The ratio of LC3-II/LC3-I of OGD/R treated RAW264.7 cells was increased (1.27 ± 0.20 vs. 0.44 ± 0.08, t = 6.67, P  < 0.05), while the expression of p62 was decreased (0.77 ± 0.04 vs. 0.95 ± 0.10, t = 2.90, P  < 0.05), and PI3K (0.40 ± 0.06 vs. 0.63 ± 0.10, t = 3.42, P  < 0.05) and p-Akt/Akt ratio was also decreased (0.39 ± 0.02 vs. 0.58 ± 0.03, t = 9.13, P  < 0.05). BM-MSCs reduced the LC3-II/LC3-I ratio of OGD/R treated RAW264.7 cells (0.68 ± 0.14 vs. 1.27 ± 0.20, t = 4.12, P  < 0.05), up-regulated p62 expression (1.10 ± 0.20 vs. 0.77 ± 0.04, t = 2.80, P  < 0.05), and up-regulated PI3K (0.54 ± 0.05 vs. 0.40 ± 0.06, t = 3.11, P  < 0.05) and p-Akt/Akt ratios (0.52 ± 0.05 vs. 0.39 ± 0.02, t = 9.13, P  < 0.05). A whole-genome microarray assay screened the differentially expressed gene HO-1, which is downstream of the PI3K/Akt signaling pathway, and the alteration of HO-1 mRNA and protein expression was consistent with the data on PI3K/Akt pathway.@*CONCLUSIONS@#Our results suggest the existence of the PI3K/Akt/HO-1 signaling pathway in RAW264.7 cells under OGD/R circumstances in vitro, revealing the mechanism underlying BM-MSC-mediated regulation of autophagy and enriching the understanding of potential therapeutic targets for the treatment of ALI.

The clinical value of shear wave elastography and salivary gland ultrasound scoring in salivary gland lesions of Sj?gren′s syndrome / 中华超声影像学杂志

Objective:To evaluate the value of shear wave elastography (SWE) and salivary gland ultrasound scoring(SGUS) system in the diagnosis of salivary glands lesions in patients with Sj?gren′s syndrome(SS) and to compare the diagnostic efficiency of the two methods.Methods:From June 2019 to November 2020, Fifty-eight patients with suspected SS were selected from the Affiliated Changzhou No.2 People′s Hospital of Nanjing Medical University. According to the 2002 American-European Consensus Group classification standard, the enrolled patients were divided into two groups: SS group (47 cases) and non-SS group(11 cases). According to symptom duration, SS group was divided into the ≤5 years group (29 cases) and the >5 years group (18 cases). Meanwhile, 40 healthy volunteers were enrolled in this study as normal control group. The diagnostic value of salivary gland ultrasound scoring system and Young′s modulus in SS were analyzed.Results:The differences in Young′s modulus of parotid gland and submandibular gland between SS group and non-SS group (or control group) were statistically significant (all P<0.05). The ultrasound score of SS group was significantly higher than that of non-SS group and control group (all P<0.05). SGUS and Young′s modulus were not significantly different between different course groups (all P>0.05). The areas under ROC curve of the mean Young′s value in parotid and submandibular gland and the SGUS were 0.801, 0.829 and 0.676, respectively. The comparison of the area under the curve between the Young′s modulus of the parotid and submandibular glands and the ultrasound score was statistically significant (all P<0.05). Conclusions:SWE and Ultrasonography scoring system have certain value in the diagnosis of salivary gland lesions in SS, and can provide important reference information for clinical diagnosis from different perspectives. The diagnostic efficiency of SWE for salivary glands lesions in patients with SS is better than that of SGUS scoring system.

Simultaneous determination of four components of Shuganning Injection in rat plasma by UPLC-MS/MS and its application to a pharmacokinetic study / 中国中药杂志

This study aims to reveal the pharmacokinetics of Shuganning Injection in normal rats. In this experiment,ultra-high performance liquid chromatography-electrospray-tandem mass spectrometry( UPLC-ESI-MS/MS) was used to establish an analytical method for simultaneous determination of chlorogenic acid,gardenioside,oroxylin A and baicalin in rat plasma. Then,the non-compartmental model( NCA) in Phoenix WinN onL in 6. 4 software was used to fit pharmacokinetic parameters. The methodological validation showed that the linear relationship of the components in rat plasma samples were good( r>0. 995). The recovery rate and matrix effect of plasma samples with low,middle and high concentration were 79. 14%-101. 4%. The intra-day and inter-day precision,accuracy and stability meet the requirements of biological sample analysis. The half-life( t1/2) of chlorogenic acid,gardenioside,oroxylin A did not change significantly and the area under blood concentration-time curve( AUC0-t) is proportional to the dose,which suggested that three components showed a linear kinetic characteristics,but baicalin showed nonlinear kinetic characteristics. Moreover,the retention time of each component in rats was short. The established UPLC-MS/MS quantitative analysis method is rapid,sensitive and accurate,which can be used for the determination of chlorogenic acid,gardenioside,oroxylin A and baicalin in rat plasma and pharmacokinetic study of Shuganning Injection.

Influences of Unidirectional Stretching Process on Mechanical Properties of Absorbable Patch for Rotator Cuff Repair / 医用生物力学

Objective To investigate the effect of unidirectional stretching on mechanical properties of different absorbable patches and evaluate its potential as a patch for rotator cuff repair. Methods The unidirectional stretching process was used to prepare absorbable patches with different polylactide based materials. Different unidirectional stretching temperatures (50-80 ℃) and stretching ratios (0.5-4.3) were set. The effects of different parameters on mechanical properties of the absorbable patches with different materials were studied. Their thermal properties, crystallization and surface morphology were characterized. Results The unidirectional stretching temperature and stretching ratio could adjust the tensile strength and strain, thermal property, crystalization and surface morphology of the absorbable patch. At directional stretching temperatures of 60, 70, 70 ℃ and stretching ratios of 3, 3, 4.3, respectively, the absorbable patches made of poly-L-lactide-co-glycolide (PLGA), poly-L-co-D, L-lactide (PLDLLA) and poly-L-lactide-co-ε-caprolactone (PLC) had the maximum tensile strength (74±7)，(97±6), (107±8) MPa, which were larger than the tensile strength for infraspinatus tendon of canine (40 MPa). However, only the strain of PLDLLA patch conformed to the flexibility of natural rotator cuff. Conclusions The unidirectional stretching process can improve mechanical properties of the absorbable patch. The absorbable patch made of PLDLLA has the potential to reinforce the rotator cuff tear.

Evaluation of Reference Genes for Quantitative PCR in Four Tissues from Rabbits with Hypercholesterolaemia

Abstract Rabbit with hypercholesterolaemia is an important model for studying cholesterol metabolism disease. This study aimed to evaluate the expression stability of nine reference genes for quantitative PCR (qPCR) analysis in adrenal gland, liver, spleen, and kidney tissue from rabbits with hypercholesterolaemia. In total, 30 male Harbin Large White (HLW) rabbits were fed a normal feed (n = 15) or a high cholesterol feed (n = 15) for 8 weeks to induce hypercholesterolaemia. Nine reference genes were verified by qPCR using cDNA extracted from rabbit tissue samples. For qPCR analysis, reference genes were evaluated using the RefFinder and GeNorm algorithms. Overall, seven rabbits with hypercholesterolaemia were identified based on body weight and total cholesterol measurements. Combining the results of the RefFinder and GeNorm algorithms, the most stable reference genes were hypoxanthine phosphoribosyltransferase 1 (Hprt1) and eukaryotic translation elongation factor 1 alpha 1 (Eef1a1) in the adrenal gland, β-2-microglobulin (B2m) and glyceraldehyde-3-phosphate dehydrogenase (Gapdh) in the liver, tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein zeta (Ywhaz) and Gapdh in the spleen, and peptidylprolyl isomerase (Ppia), β-actin (Actb), succinate dehydrogenase complex subunit A flavoprotein (Sdha), and B2m in the kidney. Taken together, our results confirmed that Hprt1 and Eef1a1, B2m and Gapdh, Ywhaz and Gapdh, and Ppia, Actb, Sdha, and B2m were the best reference genes for qPCR analyses in adrenal gland, liver, spleen, and kidney tissue, respectively, of rabbits with hypercholesterolaemia.

Degradation of silk fibroin/polylactic acid composites at different proportions in mice / 中国组织工程研究

BACKGROUND: In our previous study, we prepared some silk fibroin/polyactic acid composite materials at different proportions, which are verified as tissue-engineered materials with good performance by systemic hypersensitivity test, acute toxicity test, hemolysis test, pyrogenic test, and cell co-culture experiment. OBJECTIVE: To study the subcutaneous degradation of silk fibroin/polylactic acid composite materials at different proportions in mice. METHODS: Fifty-four ICR mice were subjected to subcutaneous cystic clearance on the both sides of the spine, and then randomized into three groups. The silk fibroin/polylactic acid composites at 40:60, 50:50, and 30:70 were implanted into the cystic space, respectively. The specimens were taken out at 1, 3, 9 weeks after implantation. Histopathological staining, Masson staining, and CD34 immunohistochemical staining were performed. RESULTS AND CONCLUSION: (1) Hematoxylin-eosin staining: At 9 weeks after implantation, new blood vessels gradually formed and visible fibroblasts gradually grew into the composites in the three groups. Structural collapse of the materials became more apparent, and tissues and cells further grew into the materials. The 40:60 and 50:50 composites were degraded more quickly than the other one. (3) Masson staining: At 9 weeks after implantation, mature blood vessels and collagen fibers were detectable in the three groups, which were more apparent in the 40:60 and 50:50 groups than the 30:70 group. The collagen fiber expression in the 50:50 group was significantly higher than that in the other two groups at different time after implantation (P < 0.05). (3) CD34 immunohistochemical staining: At 1-3 weeks after implantation, CD34 positive expression gradually increased in the three groups. After 9 weeks, the CD34 expression decreased in the three groups; however, the CD34 positive expression in the 50:50 group was significantly higher than that in the other two groups at different time after implantation (P < 0.05). To conclude, the silk fibroin/polylactic acid composite material at a proportion of 50:50 was degraded more quickly in mice.

Preclinical Study of Cell Therapy for Osteonecrosis of the Femoral Head with Allogenic Peripheral Blood-Derived Mesenchymal Stem Cells

PURPOSE: To explore the value of transplanting peripheral blood-derived mesenchymal stem cells from allogenic rabbits (rPBMSCs) to treat osteonecrosis of the femoral head (ONFH). MATERIALS AND METHODS: rPBMSCs were separated/cultured from peripheral blood after granulocyte colony-stimulating factor mobilization. Afterwards, mobilized rPBMSCs from a second passage labeled with PKH26 were transplanted into rabbit ONFH models, which were established by liquid nitrogen freezing, to observe the effect of rPBMSCs on ONFH repair. Then, the mRNA expressions of BMP-2 and PPAR-γ in the femoral head were assessed by RT-PCR. RESULTS: After mobilization, the cultured rPBMSCs expressed mesenchymal markers of CD90, CD44, CD29, and CD105, but failed to express CD45, CD14, and CD34. The colony forming efficiency of mobilized rPBMSCs ranged from 2.8 to 10.8 per million peripheral mononuclear cells. After local transplantation, survival of the engrafted cells reached at least 8 weeks. Therein, BMP-2 was up-regulated, while PPAR-γ mRNA was down-regulated. Additionally, bone density and bone trabeculae tended to increase gradually. CONCLUSION: We confirmed that local transplantation of rPBMSCs benefits ONFH treatment and that the beneficial effects are related to the up-regulation of BMP-2 expression and the down-regulation of PPAR-γ expression.

Enrichment and Biological Characteristics of Peripheral Blood-derived Mesenchymal Stem Cells in Rats / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the effective method for enrichment of rat peripheral blood-derived mesenchymal stem cells(PBMSC) and study the cell biological characteristics.</p><p><b>METHODS</b>Peripheral mononuclear cells were isolated by density gradient centrifugation from blood of 4 week old rats after G-CSF mobilization. Thereafter, the fibroblast-like cells were acquired by plastic-adherent culture, and the proliferation curve was assayed. For analyzing surface markers of the second generation cultured isolated PBMSC, both flow cytometry(CD90, CD44, CD29, CD45, CD11b and CD79a) and immunocytochemical staining(CD73, CD105, CD34 and HLA-DR) methods were used. Furthermore, the differentiation capacities of PBMSC into osteocytes, chondrocytes and adipocytes were identified.</p><p><b>RESULTS</b>(1) The adherent cells displayed typical colony-forming unit fibroblast(CFU-F) growth pattern after 6-7 day of primary culture and reached 80% confluence after 21 days of culture. The passaged PBMSC possessed high proliferative capacity and spindle growth pattern and was able to grown into exponential phase next day with a doubling time of 39.2 h. (2) PBMSC expressed mesenchymal markers such as CD90, CD44, CD29, CD73 and CD105, but failed to expressed markers of CD45, CD11b, CD79a, CD34 and HLA-DR. (3) After 21 days of culture in osteogenic, chondrogenic and adipogenic differentiation media, calcifying nodules, intracellular glycosaminoglycans and lipid droplets could be found by alizarin red, alcian blue and oil red-O staining, respectively.</p><p><b>CONCLUSION</b>PBMSC can be enriched from rat peripheral blood with high purity and abundance by our methods. The growth and phenotypic characteristics of the isolated PBMSC are consistent with that of well-known MSC, and these cells possess the capability to multi-lineage mesoderm differentiation.</p>

A case report of a bronchial foreign body removal by direct laryngoscopy with fiber bronchoscope / 临床耳鼻咽喉头颈外科杂志

An 11 year old girl presented with dyspnea and a rough cough, after having mistakenly swallowed a steel ball and unable to relieve the symptoms. Chest X-ray showed an image consistent with a 10 mm diameter circular object, embedded in the right mainstem bronchi near the fifth thoracic level and the medicastinal moving to the right. A clinical diagnosis based on these findings: foreign body of the right bronchial.

Transplantation of human amnion epithelial cells improves learning and memory function in Alzheimer’ s disease-like pathology rat model / 中国病理生理杂志

[ ABSTRACT] AIM:To observe the treatment effect and its immune regulation of human amnion epithelial cells ( hAECs) on Alzheimer’ s disease ( AD)-like pathology rat model.METHODS: The hAECs were isolated from amnion with trypsin digestion, and the phenotype of hAECs was analyzed by flow cytometry.SD rats ( n=48) were randomly divid-ed into sham control group, model group, medium group and hAECs group.AD-like pathology rat model was induced by bilateral intraventricular injection of lipopolysaccharide (LPS).hAECs (5 ×105) were injected into the hippocampus of the AD-like pathology rats.At 2 weeks after transplantation, the animals were tested by Morris water maze to observe the function of learning and memory.The pathological change of the brain was observed by HE staining.The expression of am-yloid β-protein 42 (Aβ42) and Tau protein and the level of acetylcholine (ACh) in the injury brain were determined by immunohistochemistry.The survival and differentiation of hAECs in the hippocampus were measured by immunofluorescent technique.The percentages of lymphocyte subsets in the peripheral blood mononuclear cells were analyzed by flow cytome-try.The contents of serum cytokines were detected by cytometric bead array.RESULTS:Compared with model group and medium group, hAECs group showed shortened escape latency ( P<0.01) , increased frequency of going through the plat-form (P<0.05), reduced loss of hippocampal neurons, decreased expression of Tau protein and Aβ42 in the hippocampus (P<0.05), increased ACh level in the hippocampus (P<0.05), decreased percentages of Th1 and Th17 subsets, in-creased percentages of Th2 and Treg cells ( P<0.05) , decreased concentrations of IFN-γand IL-2 in the serum, and in-creased concentration of IL-4 ( P<0.05 ) .CONCLUSION: hAECs improve the cognitive learning and memory function and alleviate pathologic damage of hippocampus through immune regulation in AD-like pathology rats.

Effects of preconditioning with phosphodiesterase-5 inhibitor on the Biological Properties of human amniotic mesenchymal stem cells / 实用医学杂志

Objective To investigate the biological properties of human amniotic mesenchymal stem cells (hAMSCs) which were preconditioned with phosphodiesterase-5 inhibitor (Vardenfil). Methods hAMSCs were in vitro isolated and cultured, hAMSCs were pre-treated with vardenfil in final concentration of 10 μmol/L. The morphology of Vard-hAMSCs was observed, and the immunological characteristics, proliferative capacity, and ability of anti-oxidative damage of hAMSCs and Vard-hAMSCs were analyzed by flow cytometry. Double labeling immunofluorescent staining was used to count the differences of differential potential between neural cells of hAMSCs and Vard-hAMSCs. Results (1)Flow cytometry revealed that both hAMSCs and Vard-hAMSCs positively expressed CD90、CD105 and CD73, and negatively expressed CD34、CD45、CD11b and HLA-DR. The SPF and PI in Vard-hAMSCs group were (0.57 ± 0.40)% and (2.20 ± 1.60)% respectively, there was no statistical significance compared with hAMSCs group; (2)After 4 hours treated by H2O2, the apoptosis rate in Vard-hAMSCs group were (7.67 ± 0.82)%,which were markedly lower than that in the hAMSCs group and specific blocker group; (3)Under the same induction condition, positive rates of MAP-2 and GFAP in Vard-hAMSCs group were (49.8 ± 6.42)%and (55.2 ± 6.10)% respectively detected by double labeling immunofluorescent staining, which were significantly higher than the control group. Conclusion The strategy that hAMSCs are treated with vandenfil can enrich the ability of anti-oxidative damage and the differential potential for neural cells in a certain time, and the morphology, immunological characteristics, proliferative capacity of Vard-hAMSCs have no significant change. It suggests that pre-treatment with vandenfil may provide a optimized experimental strategey for hAMSCs which were used to treat nervous system disease.

Clinical analysis of 56 gestational hypertension patients with heart disease / 中国基层医药

Objective To investigate the early diagnosis and preventive measures for gestational hypertensive patients with heart disease.Methods Clinical data of 56 gestational hypertensive patients with heart disease were retrospectively analyzed.Results Aggressive treatment of maternal deaths occured in 1 case,2 cases of stillbirth,and the other 55 cases had good maternal prognosis.Conclusion Treatment of hypertensive heart disease in pregnancy should be based on the strong heart vasodilator,diuretic,caution magnesium sulfate.In the control of heart failure after 6-8h,shall terminate the pregnancy.Active treatment of the primary disease,to correct anemia and hypoproteinemia,early diagnosis of heart failure and strictly control expansion of indications can help to improve the prognosis of hypertensive heart disease in pregnancy.

Therapeutic effect and immunologic regulation of human amniotic mesenchymal stem cells in rats with experimental type 1 diabetes mellitus / 中华内分泌代谢杂志

Objective To study the therapeutic effect and immunologic regulation of human amniotic mesenchymal stem cells (hAMSCs) in rats with experimental type 1 diabetes mellitus (T1DM).Methods The hAMSCs from human amnion were isolated and cultured in vitro,then phenotype was analyzed by flow cytometry.T1DM were produced by administering streptozocin to rats.The rats were divided into normal control group (n =6),T1 DM model group (n =6),medium treated group (n =6),hAMSCs transplanted group(n =6),and insulin treated group(n =6).5 × 106of hAMSCs or vehicle were administered to rats via sublingual vein.Blood glucose levels of rats were recorded weekly in the groups for six weeks by Blood Sugar Meter.At the end of 6 weeks after hAMSCs transplantation,concentrations of plasma insulin were detected by ELISA; histopathological changes of pancreas,surviwl and differentiation of transplanted hAMSCs in pancreatic tissue were studied with HE staining,and immunofluorescence staining; percentages of lymphocyte subsets in peripheral blood mononuclear cells were determined by flow cytometry ; concentrations of plasma cytokines were determined by cytometric bead array.Results After hAMSCs transplantation,blood glucose levels in rats with T1DM were decreased (P ＜ 0.01),while concentrations of plasma insulin were increased significantly (P＜0.01).At 6 weeks,cell-treated animals showed an improvement in pancreas damage ; the percentages of CD4 + IFN-γ+ (Th 1) and C D4 + interleukin (IL)-17 + (Th 17) cells were reduced (all P＜0.05),while the percentages of FoxP3-positive regulatory T cells (FoxP3 +Treg) and CD4+ IL-4+(Th2) cells were increased (all P＜0.01) ; plasma concentrations of interferon-γ,IL-2,and tumor necrosis factor-αwere decreased (all P＜0.01),but IL-4 level was increased (P＜0.05).No histological evidence of insulin producing cells from hAMSCs was seen within pancreas.Conclusions hAMSCs may reduce blood glucose and alleviate the islet damage in rats with T1 DM,which is related to their potential to up-regulate FoxP3 +Treg cells.

Verification of multipotential mesenchymal stem cell presence in peripheral blood of rabbits / 中国实验血液学杂志

The aim of this study was to verify the presence of multipotential mesenchymal stem cells in peripheral blood (PBMSC) of rabbits. For mobilization, granulocyte-colony-stimulating factor 30 µg/(kg·d) was injected into New Zealand White rabbits subcutaneously for 6 d, then the PBMSC were isolated from peripheral blood of rabbits by density gradient centrifugation and adhesive culture. The morphology of cell proliferation was observed by microscopy, the proliferative curve of cells was drawn. The phenotypes of PBMSC were detected by flow cytometry, the differential capability of PBMSC into osteocytes, chondrocytes and adipocytes was identified. The results showed that the morphology of subcultured PBMSC were spindle or polygonal shaped, and cell population doubling time was 37.4 h. The isolated PBMSC expressed mesenchymal marker CD29, but not expressed hematopoietic marker CD14. Under specific induction conditions, PBMSC demonstrated multipotency to differentiate into osteocytes, chondrocytes and adipocytes. It is concluded that PBMSC are successfully isolated from peripheral blood and cultured, and their multipotential capability of differentiation into osteocytes, chondrocytes and adipocytes are verified.

Preparation of cytoplasts from HL-60 cells / 生物医学工程学杂志

This experimental research was aimed to establish an optimum system of enucleation, purification and identification for preparing the cytoplasts of suspension culture cells in order to undertake cell recombination. Human leukemia HL-60 cells in suspension culture were purified by 42% Percoll density gradient centrifugation and low-speed centrifugation at 1 500r/min, respectively. The purified HL-60 cells were treated with cytochalasin B (CB) alone or combined with colcchicine and enucleated by isopycnic gradient centrifugation on 50% Percoll at 25 degrees C and 34 degrees C, respectively. Cytoplasts made from HL-60 cells were purified through gradient centrifugation by 37%, 38% and 40% Percoll, respectively. The final cytoplasts were identified by Wright-Giemsa staining and 4,6-diamidino-2-phenylidole dihydrochloride (DAPI)/5, 6-carboxyflu-orescein diacetate succinimidyl ester (CFSE) double-staining. The phenotype and mitochondrial membrane potential of HL-60 cytoplasts were analyzed by flow cytometry. The results indicated that the enucleation ratio of HL-60 cells induced by CB combined with colcchicine was up to 91. 98% +/-4. 29%, which was significantly higher than that in CB alone group (74. 95% +/- 3. 02%)(P<0. 01). The rates of enucleation and cytoplast with diameter over 5min in 34 degrees C group were higher than those in 25 degrees C group (all P<0. 01). The cytoplast purities were (95.43 +/- 0. 59)% in 38% Percoll groups,which were higher than those of 40% Percoll (P<0. 05). Nucleus and caryoplasm could be clearly distinguished by DAPI and CFSE double labeling. The results further showed that the phenotype of HL-60 cytoplasts had no significant change, and the activity of the cytoplasts was above 80% within 12h. It is concluded that enucleation throuth density gradient centrifugation on 50% Percoll mediated by CB combined with colcchicine, 38%Percoll of purification followed by DAPI/CFSE double labeling and MMP detection is an optimum scheme for preparation and identification of cytoplast from suspension culture cells.

Human amnion-derived mesenchymal stem cells transplantation improves hematopoietic function of myelosuppression mice / 中国组织工程研究

10.3969/j.issn.2095-4344.2013.23.010

Study on inducing differentiation of human amniotic epithelial cells into insulin secreting cells in vitro / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To investigate the differentiation of human amniotic epithelial cells (hAECs) into insulin secreting cells (ISCs) in vitro.</p><p><b>METHODS</b>The hAECs were isolated from human amnion by trypsin digestion, and the phenotype of the isolated cells were identified by flow cytometry and immunocytochemical staining. The hAECs at passage 3 were treated with nicotinamide and N2 supplement to investigate their differentiation into ISCs. At different times after differentiation, the expression of insulin and beta2 microglobulin (beta2-MG) was determined by immunocytochemical staining, while the content of insulin in supernatant from cultured hAECs was detected by radioimmunoassay and the expressions of insulin, pancreatic and duodenal homeobox factor-1 (PDX-1) mRNA were detected by reverse transcriptase-polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>(1) hAECs expressed high percent of CD29, CD73, CD166 and CK19. (2) At 7, 14 and 21 days, the percentages of insulin-positive cells in induced groups were 74.00% +/- 1.73%, 75.33% +/- 1.15% (see symbol) 75.67% +/- 0.58% respectively, which were negative in control groups. (3) At 7, 14 and 21 days, contents of insulin in supernatant from induced groups were (328.47 +/- 3.22) microIU/ml, (332.26 +/- 1.22) microIU/ml and (329.68 +/- 2.57) microIU/ml respectively, they were significantly higher than those in control groups (All P < 0.01). (4) PDX-1 mRNA and beta2-MG were expressed before and after the induction of hAECs, but insulin mRNA was expressed only in the induced groups.</p><p><b>CONCLUSION</b>hAECs can differentiate into ISCs, having the potential application for therapy of type I diabetes.</p>