AIM:To compare the biological characteristics , surface markers and multi-differentiation potential of the mesenchymal stem cells (MSCs) derived from the umbilical cord and bone marrow in the green fluorescent protein (GFP) transgenic mice.METHODS:Umbilical cord MSCs (UCMSCs) were isolated by collagen type II enzymatic diges-tion and bone marrow MSCs ( BMSCs) were isolated by density gradient centrifugation .The growth of the 2 types of MSCs was observed under inverted microscope .The cell proliferation was detected by determining the growth curve and MTT as-say.The Trypan blue method was performed to analyze the cell viability rate .The cell cycle and cell surface markers were measured by flow cytometry .The differentiation potentials of the 2 types of MSCs were tested by the differentiation kits to-ward adipocytes and osteoblasts .RESULTS:The UCMSCs attached to the culture surface 1 d after the isolation , and the cells showed spiral shape with notable growth and proliferation after 2 d of culture.After 3 d, the cell arrived sub-confluent and was ready for passage .BMSCs still showed circular shape and started to attach to the surface 4 d after culture .They formed the small colony shape only after 5 d with obvious proliferative potential .The cells became confluent 7 d after the culture.The original generation of cultivating UCMSCs growth curve was shown typically an “S” shape.But the BMSCs growth was slower than the UCMSCs .The cell proliferation was obvious for UC-MSCs in 3~5 d.BMSCs proliferated signif-icantly only after 7 d.The viability rate arrived more than 96%for both types of MSCs .The cell cycle of both MSCs did not show significant difference (G0/G1 phases were above 85%, P>0.05).Both MSCs positively expressed CD44, CD90 and CD105 (60.7%±2.3%) but the expression of CD45, CD19, CD14 and CD79 was negative (less than 25.6%±4.8%, P>0.05).More than 90%of the MSCs from the umbilical cord and bone marrow differentiated towards the adipocytes and osteoblasts without significant difference (P<0.05).CONCLUSION:UCMSCs have stronger ability of proliferation and multi-directional differentiation potentials .UCMSCs in GFP transgenic mice as a high-quality tracer can serve for tracking the stem cells in vivo.

Objective To explore the relationship between the volatile components in Angelicae Sinensis Radix from different regions of Gansu Province and its growing environment with metabolomics based on GC-MS. Methods The GC-MS method was used for detecting the volatile components in Angelicae Sinensis Radix from 31 different regions in Gansu province, and principal component analysis (PCA) and partial least squares (PLS) methods were used for analyzing and evaluating its relationship with the growing environment. Results The results of PCA showed that the volatile components in Angelicae Sinensis Radix from different regions in Gansu province were related to the altitude and the soil types. The PLS method could divide 31 samples of Angelicae Sinensis Radix from different regions in Gansu Province into three groups according to the difference of altitude. There were significant differences in the volatile components in the samples taken at different altitude regions. After analyzing linear loading plots from PCA and PLS, 11 charateristic components were screened out, including 7 compounds were identified by the retrieval of NIST11 database. Conclusion The volatile components in Angelicae Sinensis Radix from different regions in Gansu Province are closely related to the altitude and the soil type.

Objective:To study the diagnostic value of serum microRNA21 and tumor markers CEA, CYFRA21-1 and NSE in early non-small cell lung cancer (NSCLC).Methods:From January 2018 to March 2019, 22 patients with NSCLC in Daqing Longnan Hospital were selected as the study group, and 22 people who underwent health examination in our hospital during the same period were selected as the control group.The levels of serum microRNA21, major tumor markers and the relationship between pathological types and markers were observed and analyzed.Results:The serum levels of miRNA-21 (2.15±0.9), CEA [(34.1±4.9)ng/mL], NSE [(27.1±2.2)ng/mL], and CYFRA21-1 [(12.1±1.2)ng/mL] in the study group were higher than those in the control group( t=6.524, 27.392, 23.339, 27.685, all P=0.000). The serum levels of miRNA-21 (1.88±1.14), CEA [(30.1±19.9)ng/mL], CYFRA21-1 [(12.8±5.2)ng/mL] in adenocarcinoma patients were lower than those in patients with squamous cell carcinoma, and the level of NSE [(26.1±3.2)ng/mL] was higher than that of patients with squamous cell carcinomas ( t=1.158, 1.192, 0.423, 1.913, P=0.260, 0.247, 0.677, 0.070). The serum levels of miRNA-21 (2.58±0.96), CEA [(38.1±17.9)ng/mL], CYFRA21-1 [(16.8±6.2)ng/mL], NSE [(26.9±10.2)ng/mL] in NSCLC patients with Ⅲ~Ⅳ stage were higher than those in NSCLC patients with Ⅰ-Ⅱ stage, but only CYFRA21-1 had statistically significant difference( P<0.05), there were no statistically significant differences in the other three indicators ( t=1.478, 0.574, 2.114, 1.015, P=0.155, 0.573, 0.047, 0.322). Conclusion:The combined examination of serum microRNA-21 and tumor markers CEA, CYFRA21-1 and NSE in patients with NSCLC can effectively confirm the diagnosis, which is very important for the diagnosis, treatment and prognosis of patients.This diagnosis can be the first choice for patients with NSCLC.

Objective To investigate the trend of BMI among adults in Shaanxi Province from 2007 to 2015. Methods Data was obtained from China Chronic Disease and Risk Factor Surveillance from 2007 to 2015, in which a multistage clustering sampling was adopted to collect a provincially representative sample of adults in Shaanxi Province. BMI percentile(P5, P25, P50, P75, P95) and the prevalence of underweight, overweight and obesity was calculated with weight in each survey. Cochran-Armitage test was used to test trends across survey periods. Changes in BMI across survey years were compared by considering the sampling weight. Results The results of the surveillance indicated that the prevalence of underweight decreased while overweight and obesity increased among adults in Shaanxi province (Z=-14.70, P<0.001). We observed the highest increase in the prevalence of overweight and obesity among rural residents and residents aged from18 to 44. The mean BMI was estimated to increase 0.176(t=3.00, 95%CI：0.055-0.298， P=0.006) per year. We found no difference in overweight and obesity ( 2=0.196，P=0.459) between 2013 and 2015. Conclusions We note increases in overweight or obesity and a decrease in underweight among adults in Shaanxi Province. Those living in rural areas and aged from 18 to 44 led the highest increase in overweight and obesity.

This study was aimed to evaluate an effective and stable method for obtaining mesenchymal stem cells (MSC) and endothelial progenitor cells (EPC) from the rabbit bone marrow and to investigate the biological characteristics of MSC and EPC. Mononuclear cells were obtained from rabbit bone marrow using density gradient method, and were differentially adhered to the cell culture plate enclosed with fibronectin. Then, MSC and EPC were amplified with EGB-2MV medium. Trypan blue method was used to test the passage survival rate. Growth curve, MTT and DNA cycle were used to evaluate the proliferation ability of MSC and EPC. MSC were identified with induced differentiation into the osteoblasts and adipocytes, and their immune phenotype was detected by flow cytometry (FCM). EPC were characterized by the special digestion of Dil-ac-LDL, FITC-UEA-I, and the conjunction with CD133, VEGFR2/KDR and CD34, their purity was also calculated. The results indicated that the colony was obviously formed when the mononuclear cells were cultured for 24 hours and, 80% of the cells became long spindle and integrated at d 8. Cells, which were adhered for twice, were cultured with EGM-2MV medium, began to extend at d 3, and became strip-shaped and integrated for about 80% at d 8. Passage survival rates were more than 90% for both cells, and after passage 2 the growth curve was like "S". Optical density was changed obviously when the cells were cultured for 3 - 5 d, but there were no significant difference of cell cycles between MSC and EPC, which G0-G1 was (93.32 ± 1.65)% and (93.05 ± 1.95)% respectively. Positive rates were (99.7 ± 1.12)%, (99.1 ± 2.33)%, (4.8 ± 0.38)%, (6.8 ± 0.49)% and (0.4 ± 0.08)% for CD90, CD44, CD14, CD45 and CD79a respectively. MSC were identified by induced differentiation into osteoblasts and adipocytes. Positive rates of the EPC, which were adhered for twice and passaged 2, were (82.1 ± 3.4)% for fluorescent staining of Dil-ac-LDL and FITC-UEA-I, and (74.2 ± 3.2)%, (64.7 ± 4.3)% and (43.5 ± 1.5)% for CD133, VEGFR2/KDR and CD34 respectively. It is concluded that high-purity MSC can be obtained with density gradient and differential adhesion method, and high-proliferation EPC can be cultured with EGM-2MV medium in cell plates enclosed with fibronectin, so they may become the optimal seed cells for tissue engineering study.
Animals ; Bone Marrow Cells ; cytology ; Cell Adhesion ; Cell Differentiation ; Cell Proliferation ; Cell Separation ; methods ; Cells, Cultured ; Endothelial Cells ; cytology ; Flow Cytometry ; Immunophenotyping ; Mesenchymal Stromal Cells ; cytology ; Rabbits ; Stem Cells ; cytology

With the development of molecular biology and genomics,metagenomics is playing a more important role in forensic science and forensic identification.In recent years,as a branch discipline studying the composition profile and diversity of microbe flora as well as studying the interaction within microbe and with environment,the application of metagenomics has gradually risen and brought new opportunities for forensic identification-related area.In this review,strategy of metagenomics and its application in forensic identification including individual identification,origin determination of biological stain in crime scene and drug abuse detection are summarized.This article aims to elucidate the role and application value of metagenomics in forensic science.

OBJECTIVETo evaluate dietary iodine intake and its potential risks among the Chinese population.

METHODSIndividual dietary iodine intake was calculated using food consumption data multiplying by iodine concentration in foods, table salt and drinking water, followed by summing, and then compared with the corresponding age-specific reference values, including Upper Intake Level (UL) and Recommended Nutrient Intake (RNI).

RESULTSIn areas with water iodine concentration (WI) lower than 150 μg/L, 80.8% of residents had iodine intake between the RNI and UL, 5.8% higher than UL, and the remaining (13.4%) lower than RNI if iodized salt was consumed. However, in the uniodized salt consumption scenario, only 1.0% of residents between RNI and UL, 1.4% higher than UL, and a large part of residents (97.6%) lower than RNI. In areas with WI higher than 150 μg/L, all residents had iodine intake between RNI and UL if iodized salt was consumed, except 10.5% and 24.9% of residents higher than UL in areas with WI at 150-300 μg/L and higher than 300 μg/L respectively. However, in the uniodized salt consumption scenario, only 1.5% and 1.7% of residents had higher iodine intake than UL respectively.

CONCLUSIONThe findings suggested that in general, the dietary iodine intake by the Chinese population was appropriate and safe at the present stage. People in areas with WI lower than 150 μg/L were more likely to have iodine deficiency. While people in areas with WI higher than 150 μg/L were more likely to have excessive iodine intake if iodized salt was consumed.

Adolescent ; Child ; Child, Preschool ; China ; epidemiology ; Diet ; Drinking Water ; chemistry ; standards ; Female ; Goiter ; epidemiology ; prevention & control ; Humans ; Iodine ; administration & dosage ; analysis ; deficiency ; Male ; Nutritional Status ; Sodium Chloride, Dietary ; administration & dosage ; analysis

The proliferation of cardiac fibroblasts (CFs) is a key pathological process in the cardiac remodeling. To establish an objective, quantitative method for the analysis of cell proliferation and cell cycle, we applied the high-content screening (HCS) and flow cytometry (FCM) techniques. CFs, isolated by enzyme digestion from newborn C57BL/6J mice, were serum starved for 12 h and then given 10% fetal bovine serum (FBS) for 24 h. Followed by BrdU and DAPI (or 7-AAD) staining, CFs proliferation and cell cycle were analyzed by HCS and FCM, respectively. Discoidin domain receptor 2 (DDR2) staining indicated that the purity of isolated CFs was over 95%. (1) HCS analysis showed that the ratio of BrdU-positive cells was significantly increased in 10% FBS treated group compared with that in serum-free control group [(12.96 ± 0.67)% vs (2.77 ± 0.33)%; P < 0.05]. Cell cycle analysis showed that CFs in G0/G1 phase were diploid, and CFs in S phase were companied with proliferation, DNA replication and enlarged nuclei; CFs in G2 phase were tetraploid, and CFs in M phase produced two identical cells (2N). (2) FCM analysis showed that the ratio of BrdU-positive cells was increased in 10% FBS treated group compared with that in the control group [(11.10 ± 0.42)% vs (2.22 ± 0.31)%; P < 0.05]; DNA content histogram of cell cycle analysis indicated that the platform of S phase elevated in 10% FBS group compared with control group. (3) There were no differences between the two methods in the results of proliferation and cell cycle analysis. In conclusion, HCS and FCM methods are reliable, stable and consistent in assessment of the proliferation and cell cycle in CFs.
Animals ; Cell Cycle ; Cell Proliferation ; Fibroblasts ; cytology ; Flow Cytometry ; Mice ; Mice, Inbred C57BL ; Mitosis ; Myocardium ; cytology

Objective::To establish a pre-column derivatization reverse-phase high-performance liquid chromatography (RP-HPLC) method for the simultaneous determination of 17 amino acids in Cynomorii Herba from different producing areas and conduct a multivariate statistical analysis. Method::RP-HPLC with pre-column derivatization was employed, with phenyl isothiocyanate (PITC) as derivatization reagent. Separation was performed on a WondaSil C₁₈-WR column (4.6 mm×150 mm, 5 μm), with 0.05 mol·L^-1 sodium acetate solution (pH 6.5) as mobile phase A, and acetonitrile-methanol-water (3∶1∶1) as mobile phase B for gradient elution at a flow rate of 0.8 mL·min^-1. The detective wave length was set at 254 nm, and the column temperature was maintained at 35 ℃. Principal component analysis (PCA) and systematic cluster analysis (HCA) models were established for multivariate statistical analysis and quality evaluation. Result::17 Kinds of amino acid were detected in Cynomorii Herba, 7 of which were essential amino acids. The 17 amino acids showed good linearity in respective concentration range, r = 0.999 0-0.999 9.The average recoveries were between 98.03%-103.89%with RSD<3.5%. The results of PCA and HCA were basically the same, and both methods can be used to clearly distinguish Cynomorii Herba from 12 municipal producing areas into 6 regions. PCA can be used to classify Cynomorii Herba according to different municipal or provincial production areas, and HCA can be used to classify it according to provincial production areas. It showed that the amino acid contents in Cynomorii Herba from different municipal and provincial producing areas had differences, and the content distribution showed obvious geographical clustering characteristics. PCA showed that Cynomorii Herba from Gansu province and Inner Mongolia had higher amino acid contents and better quality as compared with other producing areas. Conclusion::The established method can be used for content determination of 17 amino acids in Cynomorii Herba from different producing areas, and provide a reference for its comprehensive quality evaluation.