BACKGROUND AND OBJECTIVEIt has been proven that epithelial-mesenchymal transition (EMT) not only correlated with embryonic development but also could promote tumor invasion and metastasis. Transforming growth factor beta-1 (TGF-beta1) has been identified as the main inducer of tumor EMT. The aim of this study was to investigate the effects of TGF-beta1 on EMT and PI3K/AKT signaling pathway in lung adencarcinoma PC9 cells.

METHODSCultured PC9 cells were treated with different concentrations of TGF-beta1 for 48 h. The morphological changes were observed under phase-contrast microscopy; EMT relative marker protein changes were assessed by Western blot and immunoflurescence staining. In addition, the expression of AKT and P-AKT were also measured by Western blot.

RESULTSThe data showed that TGF-beta1 could induce PC9 morphological alteration from epithelial to mesenchymal and upregulate the expression of mesenchymal maker protein Fibronectin. Obviously, the expression of P-AKT was downregulated by TGF-beta1 treatment for 48 h.

CONCLUSIONTGF-beta1 might induce EMT of PC9 cells, accompanied by the changes of PI3K/AKT signaling pathway.

Adenocarcinoma ; metabolism ; pathology ; Blotting, Western ; Cell Differentiation ; drug effects ; Cell Line, Tumor ; Epithelial Cells ; pathology ; Fibronectins ; metabolism ; Humans ; Lung Neoplasms ; metabolism ; pathology ; Mesoderm ; metabolism ; pathology ; Microscopy, Phase-Contrast ; Proto-Oncogene Proteins c-akt ; metabolism ; Signal Transduction ; drug effects ; Transforming Growth Factor beta1 ; pharmacology

Humans ; MicroRNAs ; genetics ; physiology ; Neoplasm Metastasis ; genetics ; pathology ; Neoplasms ; genetics ; pathology

BACKGROUND:Cartilage tissue engineering has been widely used to achieve cartilage regeneration in vitro and repair cartilage defects. Tissue-engineered cartilage mainly consists of chondrocytes, cartilage scaffold and in vitro environment. OBJECTIVE:To mimic the environment of articular cartilage development in vivo, in order to increase the bionic features of tissue-engineered cartilage scaffold and effectiveness of cartilage repair. METHODS: Knee joint chondrocytes were isolated from New Zealand white rabbits, 2 months old, and expanded in vitro. The chondrocytes at passage 2 were seeded onto a scaffold of articular cartilage extracelular matrix in the concentration of 1×106/L to prepare cel-scaffold composites. Cel-scaffold composites were cultivated in an Instron bioreactor with mechanical compression (1 Hz, 3 hours per day, 10% compression) as experimental group for 7, 14, 24, 28 days or cultured staticaly for 1 day as control group. RESULTS AND CONCLUSION:Morphological observations demonstrated that the thickness, elastic modulus and maximum load of the composite in the experimental group were significantly higher than those in the control group, which were positively related to time (P < 0.05). Histological staining showed the proliferation of chondrocytes, formation of cartilage lacuna and synthesis of proteoglycan in the experimental group through hematoxylin-eosin staining and safranin-O staining, which were increased gradualy with mechanical stimulation time. These results were consistent with the findings of proteoglycan kit. Real-time quantitative PCR revealed that mRNA expressions of colagen type I and colagen type II were significantly higher in the experimental group than the control group (P < 0.05). The experimental group showed the highest mRNA expression of colagen type I and colagen type II at 21 and 28 days of mechanical stimulation, respectively (P < 0.05). With the mechanical stimulation of bioreactor, the cel-scaffold composite can produce more extracelular matrix, such as colagen and proteoglycan, strengthen the mechanical properties to be more coincident with thein vivo environment of cartilage development, and increase the bionic features. With the progress of tissue engineering, the clinical bioregeneration of damaged cartilage wil be achieved.

BACKGROUND:Transplantation of alogeneic intervertebral disc can be facilitated by the cryopreservation of the intervertebral disc. But the traditional cryopreservation methods always lead to the appearing of ice crystals inside and outside the cels which can cause celular injury. The vitrification method that can avoid the formation of ice crystals have been widely applied in the cryopreservation field. However, only a few reports have assessed the vitrified cryopreservation of the intervertebral disc, and the toxicity of cryoprotectants to the nucleus pulposus cels have not been fuly explored.
OBJECTIVE:To determine the order of toxicity of five commonly used cryoprotectants that are used alone or in combination to rabbit nucleus pulposus cels, and to select the optimal cryoprotectant for the vitrification of the intervertebral disc.
METHODS: We chose five most commonly used cryoprotectants including dimethyl sulphoxide, formamide, ethylene glycol, propylene glycol and glycerol. Then, 5 single commonly used cryoprotectants, 10 mixed agents containing any 2 commonly used cryoprotectants, and 10 mixed agents containing any 3 commonly used cryoprotectants were formulated. Cel viability of nucleus pulposus cels was determined using cel counting kit-8 and fluorescein diacetate/propidium iodide method. Al data obtained were analyzed statisticaly to choose the appropriate combining scheme with less toxicity.
RESULTS AND CONCLUSION: The order of the toxicity of these five commonly used cryoprotectants from low to high was ethylene glycol, glycerol, formamide, dimethyl sulphoxide, and propylene glycol. The toxicity of the combined agents containing two or three commonly used cryoprotectants was lower than that of any commonly used cryoprotectants that were used to formulate them. The toxicity of the mixed agents that contained ethylene glycol or glycerol was lower than that of any other mixed agents. So we can choose the mixed cryoprotectants that contain ethylene glycol and (or) glycerol for the vitrification of the intervertebral disc.

BACKGROUND:Magnesium can be degraded voluntarily in vivo, so a second surgery is avoided. However, its aloys have not been widely used in the clinical orthopedics because there is a lack of accurate and reliable methods to assess its degradationin vivo.
OBJECTIVE:To explore the degradation of micro-arc-oxidized AZ31 magnesium aloy in the femoral condyle of rabbits based on micro-CT images and relative data.
METHODS:Forty micro-arc-oxidized AZ31 magnesium aloys were implanted into the right femoral condyle of 40 New Zealand rabbits. Then 10 right femoral condyles were removed at 5, 10, 15 and 20 weeks after surgery, respectively, to quantitatively analyze and evaluate the degradation of AZ31 magnesium aloys by micro-CT images and relative data.
RESULTS AND CONCLUSION:The surface of AZ31 aloys was corroded progressively with dark color and distorted appearance at 5-20 weeks post implantation. Micro-CT images showed that in the first 5 weeks, the degradation was inactive, and at the 10th week, it turned active; at the 15th week, the corrosion pits were obviously increased in number, and the corrosion area and corrosion speed were enlarged and fastened, respectively. Up to the 20th week, the aloy surfaces were ful of corrosion pits besides roughness and discontinuity. Relevant data analysis showed that the volume fraction of magnesium aloy was 98.6%, 97.1% and 86.4% at the 5th, 10th and 20th weeks after implantation, respectively, and it had a significant decrease from the 10th to 15th week and from the 15th to 20th week (P < 0.05). Within 15-20 weeks, the volume fraction of magnesium aloy was decreased by 6.5% that was the maximum volume reduction per unit cycle. With the progress of corrosion, the surface continuously became rough and vague, and its surface area was enlarged; the ratio of surface area to volume continuously increased, and there was a significant difference at 15 and 20 weeks (P < 0.05). Because of the increasing number of corrosion pits, the cross-sectional radius decreased, which was reflected by the trabecular thickness decreasing from 1.00 to 0.87 mm. From the view of the slope of curve, the trabecular thickness decreased most rapidly at 10-15 weeks. The mineral density of magnesium aloy continuously decreased from 649.302 to 356.445 mg/cm3 during the whole experiment period (P< 0.05). In addition, the micro-CT image density decreased from 679.710 to 644.947 mg/cm3, but there was no significant difference. To conclude, the degradation speed is peaked at 10-20 weeks after implantation, and the content of magnesium aloys decrease with degradation, but the magnesium density has no significant change.

The present paper was aimed to study the relationship between the pneumonia clinical features and the pathogens of pneumonia in children by making use of association rules based on the clinical data of 6 300 cases of pneumonia. Through software analysis, the different association relationship can be obtained between different clinical features of pneumonia in children, such as gender, age and region, etc., and the pathogens of pneumonia. For example, children of different sex with the same pathogen showed different association relationships. Due to the different association relationships between the pneumonia clinical features and the pathogens of pneumonia in children of Guangzhou area, different methods in prevention and treatment of children's pneumonia should be adopted according to actual condition, in order to achieve the best results.
Adolescent ; Age Factors ; Bronchopneumonia ; epidemiology ; microbiology ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Incidence ; Infant ; Male ; Pneumonia ; epidemiology ; microbiology ; Pneumonia, Mycoplasma ; epidemiology ; microbiology ; Pneumonia, Viral ; epidemiology ; microbiology ; Sex Factors ; Software

Objective To explore the impact of early use of renal replacement therapy on the prognosis of severe acute pancreati-tis patients .Methods 60 cases of patients with severe acute pancreatitis were chosen between September 2015 and June 2016 in De-partment of General Surgery in our hospital ,and were randomly divided into treatment group (n=30) and control group (n=30) with the method of number table .The patients in the control group were treated with conventional therapy ,treatment group was given the early use of renal replacement therapy (within 12 h after onset) .The scores of Acute Physiology and Chronic Health E-valuation Ⅱ (APACHE Ⅱ) ,partial pressure of oxygen (PaO2 ) ,oxygenation index (PaO2/FiO2 ) ,lactate (Lac) ,C-reactive protein (CRP) ,interleukin 1 (IL-1 ) ,and interleukin-6 (IL-6) levels before treatment ,12 h ,24 h ,48 h and 72 h after treatment were com-pared between two groups ,and the incidence of abdominal compartment syndrome (ACS) was compared as well .Results The difference of the indicators before treatment between two groups was not significant different (P>0 .05);the scores of APACHEⅡ ,PaO2 ,PaO2/FiO2 and Lac levels at 12 h ,24 h ,48 h and 72 h after treatment in treatment group were all lower than those in con-trol group ,and the difference was statistically significant (P<0 .05);CRP ,IL-1 and IL-6 levels at 24 h ,48 h and 72 h after treat-ment in treatment group were lower than those in control group ,and the difference was statistically significant (P<0 .05) .In the control group ,5 cases had ACS ,the incidence rate was 16 .7% ,and there was no case having ACS in treatment group and the differ-ence was statistically significant (P<0 .05) .Conclusion The early application of renal replacement therapy can effectively improve the physiological indexes of patients with severe acute pancreatitis ,inhibit the inflammatory response rapidly ,reduce damage to the abdominal organs ,and is worthy of promotion and application in clinical practice .

Objective:To analyze the relationship between homology of Kleber pathogen pneumoniae (KP) in patients with neurocritical infections and the Genomics.Method:Five non-multidrug resistant pathogen KP were identified in 2015 to 2018, including the same cloning strain of P90 and P91, the same popular cloning system of P66,P90 and P91, and there is no homology between P20,P39 and other strains, which makes a second generation full genome sequencing. A variety of bioinformatics software were used for genomic analysis to understand the basic genomic information, chromosomal and plasmid distribution, single nucleotide polymorphism (SNP) differences and gene family clustering characteristics, meanwhile with the National Center for Biotechnology Information (NCBI) website registered 18 KP strains (2013--2016) to analyze the evolutionary affinity between strains.Results:The total genome sizes of P20, P39, P66, P90 and P91 were 5 469 543 bp, 5 480 332 bp, 5 768 352 bp, 5 745 666 bp, 5 722 999 bp. The GC contents were 57.07% (1 559 929+1 561 432)/5 469 543, 57.27% (1 566 970+1 571 424)/5 480 322, 56.96% (1 640 438+1 645 432)/5 768 352, 56.88% (1 634 285+1 634 038)/5 745 666, and 56.95% (1 627 360+1 631 781)/5 722 999, respectively. Compared with P20 reference strains, the total number of SNP in P39, P66, P90 and P91 were 32 682, 34 226, 34 292, 34 375, and the total mutation rates of gene coding region sequences were87.18% (28 491/32 682), 86.71% (29 679/34 226), 85.26% (29 238/34 292), 86.22% (29 638/34 375), respectively. Nonsynonymous mutations accounted for some advantages, and the rates were 44.57% (14 566/32 682), 44.01% (15 063/34 226), 48.01% (16 465/34 292), 48.75% (16 758/34 375), and synonymous mutations were 42.61% (13 925/32 682), 42.70% (14 616/34 226), 37.25% (12 773/34 292), 37.47% (12 880/34 375), respectively. P90 and P91 have 6 specific gene families, and P66 has 4 specific gene families. The same popular clone lines P66, P90 and P99 are on the same evolutionary branch of the phylogenetic tree. The same clone P90 and P99 are on the same subbranch. P20 and P39 without homology are on different evolutionary branches respectively. P20, P39, P66, P90 and P91 on the evolutionary branches of phylogenetic tree are closely related to the evolutionary grade of strain KP52-145 from France and strain ED23 from Taiwan, China submitted on NCBI website.Conclusion:Klebsiella pneumoniae in patients with neurocritical infection has the same clone, and the number of unique gene families among strains is the same. There are small differences in the number of unique gene families and the total number of SNPs among the same epidemic clone lines, and they are characteristic of the same evolutionary branch of the phylogenetic tree. The number of unique gene families and the total number of SNPs of non homologous strains are quite different, and they are in different evolutionary branches of the phylogenetic tree.

OBJECTIVETo assess the value of combined fetal karyotyping and chromosomal microarray analysis (CMA) for the verification of high-risk pregnancy signaled by noninvasive prenatal screening (NIPS) based on high-throughput sequencing.

METHODSOne hundred and fifty-one pregnant women with high risks for aneuploidies of chromosomes 13, 18, 21, X and Y or pathological copy number variations (CNVs) by NIPS were subjected to amniocytic karyotyping and CMA analysis.

RESULTSOne hundred and forty-two women were found to have a high risk for fetal chromosomal aneuploidies, which included 83 cases of trisomy 21, 17 cases of trisomy 18, 2 cases of trisomy 13, and 40 cases of sex chromosome aneuploidies. Amniocytic karyotyping and CMA analysis has confirmed 81 cases of trisomy 21, 15 cases of trisomy 18, 10 cases of 47,XXY, 4 cases of 47,XXX, 2 cases of 47,XYY and 1 case of 46,X,del(X)(q26.1). Two trisomy 21, two trisomy 18, 2 trisomy 13, and 23 cases of sex chromosomal aneuploidies were verified as false positives. For 9 women with pathological fetal CNVs detected by NIPS, combined fetal karyotyping and CMA has confirmed 1 case of chromosome 13 microdeletion, 1 case of chromosome 18 microduplication, and 1 case of chromosome 18 deletion. For a case with 30 Mb duplication of chromosome 2 and 25 Mb duplication of chromosome 8, CMA analysis had no positive finding, while fetal umbilical cord blood karyotyping has yielded a 46,XX,dup(2)(p23.1p25.3)[13]/46,XX[87] karyotype. The remaining 5 cases were confirmed as false positive results.

CONCLUSIONCombined fetal karyotyping and CMA has provided a powerful tool for verifying high-risk fetuses signaled by NIPS.

Aneuploidy ; DNA Copy Number Variations ; Down Syndrome ; Female ; High-Throughput Nucleotide Sequencing ; methods ; Humans ; Karyotyping ; Microarray Analysis ; Pregnancy ; Prenatal Diagnosis ; Trisomy 13 Syndrome ; Trisomy 18 Syndrome