BACKGROUND:Research has been proved that Fasudil,a Rho kinase inhibitor,can effectively inhibit the onset of secondary spinal cord injury.OBJECTIVE:To investigate the synergistic effect of bone marrow masenchymal stem cell (BMSC) transplantion and Fasudil on motor functional recovery following spinal cord injury.DESIGN,TIME AND SETTING:A randomized controlled animal experimant was performed at institute of Endocrinology,Tianjin Medical University from November 2008 to March 2009.MATERIALS:A 1-month-old SD rat was obtained to extract BMSCs.Another 30 healthy female SD rats were used to establish spinal cord injury models,and they were then randomly divided into single injury group,cell transplantation group,and cell transplantation + Fasudil group,with 10 rats for each group.Fasudil was provided by Tianjin Hongri Pharmaceutical Co.,Ltd.METHODS:One week after modeling,spinal cord injury was exposed in the cell transplantation group and cell transplantation+Fasudil group,and 10 μL BMSC suspension was inserted into the injured region.Otherwise,6 hours later rats in the cell transplantation +Fasudil group were treated with an intraperitoneal injection of 10 mg/kg Fasudil,twice a day for one successive week.MAIN OUTCOME MEASURES:Hindlimb motor function was detected using inclined plane test.The Phosphorylated-ERM protein expression was detected by hernatoxylin-eosin staining,pathology and horseradish peroxidase (HRP) nerve trace,and Western blot.RESULTS:Eight weeks after modeling,degree of inclined plane was significantly increased in cell transplantation group and cell transplantation + Fasudil group compared with single injury group (P ＜ 0.05,P ＜ 0.01);while the increased value in the cell transplantation group was significantly greater than cell transplantation + Fasudil group (P ＜ 0.05).Broken myeloid tissue and cavitation were observed in the single injury group;a few of neuraxis-like structures were observed in the cell transplantation group and cell transplantation + Fasudil group,but the cavity in the cell transplantation group was larger than cell transplantation + Fasudil group.HRP-pesitive nerve fibers were detected at T_8 segment or even above in the single injury group and increased in cell transplantation group,in particular in cell transplantation + Fasudil group.Phospho-ERM protein expression in the single injury group and cell transplantation group was significantly greater than cell transplantation + Fasudil group (P ＜ 0.05).CONCLUSION:BMSC transplantation can promote hindlimb motor functional recovery following spinal cord injury,while the combined application of cell transplantation and Fasudil may cause a synergistic effect.

Objective To study dynamic changes of gene expressions and protein synthesis of Runx2 (runt-related transcription factor-2), Osterix, osteocalcin and AJ18 inside the femoral head with steroid-induced osteonecrosis after mechanical stress stimulation in rats. Methods A total of 50 Wistar rats (half male in sex) weighing 250-270 g (mean 260 g) were involved in this study and randomly divided into experimental group (40 rats) and normal group (10 rats). The rats in experimental group were injected with dexamethasone (20 mg/kg) via bilateral gluteus maximus alternatively once a week and then trained on laboratory animal treadmill twice weekly to make rat model of femoral head necrosis. After identifying the successfully induced model by Hematoxylin and eosin stain, glucocorticoid injection was ceased and the experimental group was randomly divided into model control group, 4 weeks, 6 weeks, 8 weeks groups after hormone training stopped. Then, total RNA and total protein were extracted from femoral head for detecting dynamic changes of genes expressions and proteins synthesis of Runx2, Osterix, osteocalcin and A J18 after mechanical stress stimulation inside the femoral head with steroid-induced osteonecrosis by means of real-time quantitative PCR and Western blot assay. Results In 4 weeks, 6 weeks, 8 weeks groups after hormone training stopped, the gene expressions and proteins synthesis of Runx2, Osterix and osteocalein were reduced more significantly compared with model control group, mBNA expression values of Runx2, Osterix and osteoealcin were 0. 1809, 0. 1639, 0. 1374 and 0. 4219, 0. 3026, 0.2652 and 0. 2857, 0.2027, 0. 1583 times of those in model control group. The expressions of Runx2, Osterix and osteocalcin showed a downward trend with time. The mBNA expression and protein synthesis of AJ18 at 4th, 6th and 8th weeks after hormone training stopped were 2.6391,4. 2718 and 5. 3165 times of model control group. Conclusions In addition to hormonal factors, inappropriate mechanical stress inhibits expressions and proteins synthesis of Runx2, Osterix and osteocalcin, while the expression and protein synthesis of AJ18 are upgraded in early steroid-induced femoral head necrosis in rats.

Objective To investigate the relationship between blood glucose levels and severity of coronary stenosis in hypertensive patients. Methods Blood pressure, fasting plasma glucose, 2 h plasma glucose, clinical features and coronary angiographic findings were analyzed retrospectively in 540 patients with essential hypertension. Acoording 2 h plasma glucose, patients were stratified into three group: group 1: 2h plasma glucose

Objectives:To establish hybridomas secreting monoclonal antibodies(McAbs) against O6-methylguanine-DNA methyltransferase(MGMT) and to observe the relationship between MGMT expression and clinical responses to ACNU and BCNU in human brain tumors.Methods:The hybridomas were established by cell fusion.MGMT expression in 60 glioma specimens was detected by means of immunohistochemical assay.Results: Seven hybridomas secreting McAbs against MGMT were obtained.Thirty tumor specimens had no detectable or low level of MGMT expression(Mer-), while 30 specimens had high level of MGMT expression(Mer+). The Mer- patients showed more sensitive to ACNU and BCNU than the Mer+ patients.Conclusions: The high specific hybridomas secreting monoclonal antibodies(McAbs) against MGMT were established.The preliminary study indicated that MGMT negative tumors were sensitive to ACNU and BCNU, whereas MGMT positive ones were more resistant to nitrosourea drugs.

Objective To verify the genetic characteristics associated with gastric cancer,and to propose a hybrid feature selection method for identifying target genes,further analyzing their significance and establishing a new diagnostic prediction model.Methods Analysis of variance in bioinformatics was performed on the original gastric cancer data,and then machine learning methods such as random forest,recursive feature elimination of support vector machine,and LASSO algorithm were used to screen gastric cancer associated genes,and the intersection of results was taken as the key gene set.The key genes were identified and verified through enrichment analysis.The diagnosis and prediction models based on 8 kinds of machine learning classification algorithms such as multi-layer perceptron,logistic regression and decision tree,were constructed using the key genes.Results The key genes selected by the hybrid feature selection method were closely related to the tumorigenesis and development.Eight key genes(TXNDC5,BMP8A,ONECUT2,COL10A1,JCHAIN,INHBA,LCTL and TRIM59)were identified as potential markers of good diagnostic efficacy in gastric cancer.The ROC curve and accuracy results demonstrated that among the 8 classification models,MLP is the best gastric cancer prediction model,with an accuracy of 97.77%,which was 3.83%higher than that of Xgboost gastric cancer prediction model.Conclusion The study identifies 8 key genes for the diagnosis and prevention of gastric cancer,and establishes the optimal prognosis model.

Tumors are serious diseases threatening human health,and the early diagnosis is essential to improve treatment success and patient survival.The study of tumor gene expression data has become a major tool for revealing tumor disease mechanisms,in which artificial intelligence plays an important role.The potential advantages of supervised learning,unsupervised learning and deep learning in tumor prediction and classification are explored from the perspective of machine learning methods.Special attention is paid to the impact of feature selection algorithms on gene screening and their importance in high-dimensional gene expression data.By providing a comprehensive overview of the application and development of artificial intelligence in the analysis of tumor gene expression data,the study aims to provide an outlook for future research directions and promote further development.

Objective:To investigate the mechanism of Wenjing Decoction in the treatment of hepatic fibrosis through network pharmacological methods, and conducting animal experiments to verify the core targets.Methods:The TCMSP database platform was used to screen the active components and related targets of Wenjing Decoction, and the Uniprot database was used to obtain the target genes corresponding to the active components of Wenjing Decoction. The network diagram of "Chinese materia medica-compound-target" was constructed in Cytoscape 3.7.2, and the GeneCards database was used to search liver fibrosis related targets. String database was used to construct a protein interaction network (PPI) to screen the core components and key targets of liver fibrosis, and GO analysis and KEGG pathway enrichment were performed. Animal experiments were conducted to verify the results of the analysis. 10 mice were selected as the blank group, and the remaining 45 rats were induced with carbon tetrachloride induced liver fibrosis model. After modeling, 40 successfully modeled rats were divided into model group and Wenjing Decoction high, medium-, and low-dosage groups using a random number table method, with 10 rats in each group. Wenjing Decoction high, medium-, and low-dosage groups were orally administered with 1.5×3.18, 3.18, and 0.5×3.18 g/kg Wenjing Decoction, respectively. The blank group was orally administered with equal volume distilled water once a day for 8 consecutive weeks. HE staining was used to observe the histopathological and morphological changes in the liver of rats. The serum GPT and GOT levels of rats were detected using a fully automated biochemical analyzer, and the expressions of TNF, AKT, and IL-6 proteins in rat liver tissue was detected using Western Blot.Results:A total of 188 active components of Wenjing Decoction were obtained, and the active components with higher degree values were β-sitosterol, quercetin, naringenin, etc. 799 liver fibrosis gene targets were collected, and the core target genes of the PPI network were TNF, AKT, IL6, etc. The key anti-hepatic fibrosis related pathways were obtained by GO function and KEGG analysis, including pathway in cancer, TNF, PI3K-Akt and other signalling pathways. Results of animal experiments showed that there were obvious inflammatory infiltration, collagen fibre and pseudo lobe generation in the liver tissue of rats in the model group, and the levels of inflammation and fibrosis in the liver tissue of rats in the Wenjing Decoction high, medium-, and low-dosage groups were improved to different degrees compared with that of the model group; compared with the model group, the levels of serum GPT and GOT decreased ( P<0.05); the protein expressions of TNF, AKT and IL6 in the Wenjing Decoction high, medium-, and low-dosage groups decreased ( P<0.05). Conclusion:Wenjing Decoction may exert anti-liver fibrosis effects by intervening in TNF, AKT, IL6 targets, regulating cancer pathways, TNF, PI3K Akt and other signaling pathways.

Objective To evaluate the application value of ultrasound in vascularization of different artificial bones . Methods A total of 15 New Zealand rabbits were utilized for model establishment of classic segmental bone defect in bilateral radius . Recombinant human bonemorphogenic protein-2 ( rhBMP-2 ) coralline hydroxyapatite(CHA) and CHA were implanted into left and right limbs . Each CHA was divided into 4 equal parts which were examined with conventional ultrasonography and contrast-enhanced ultrasonography( CEUS ) on 3 d ,7 d ,11 d ,15 d ,30 d and 45 d respectively . CEUS quantitative was performed by time-intensity curve(TIC) ,which parameters including the basic intensity(BI) ,peak intensity (PI) ,increased signal intensity ( ΔSI) and time to peak ( TTP) . Then the results were analyzed and compared to pathology . Results Within the same duration ,the vascularization degree in rhBMP-2 group was stronger than that in the ordinary group with advanced vascularization time . Positive correlation was detected between ΔSI and time of both groups ( r =0 .938 ,0 .890 ;P =0 .000) ,and negative correlation was found between BI/PI or TTP and time ( BI/PI: r = -0 .798 ,-0 .899 ; P = 0 .000 ;TTP= r -0 .874 ,-0 .868 ;P = 0 .000 ) . No statistical significance was observed among four observation points of both CHA ,which indicated no obvious difference in vascularization degree of each observation point . Conclusions The structure of bone graft can be clearly displayed by conventional ultrasound ,and CEUS is able to show the early blood perfusion in two CHA grafts and to accurately evaluate the difference of CHA microvascular growth before and after rhBMP-2 application . The combination of these two techniques is a promising approach of evaluating bone graft vascularization in clinical practice .

OBJECTIVETo determine the origin of a supernumerary small marker chromosome found in a fetus using prenatal BACs-on-Beads (BoBs) and single nucleotide polymorphism array (SNP-array) assays.

METHODSThe fetal sample was subjected to chromosomal karyotyping and BoBs analysis, and the results were validated with genome-wide scanning using a SNP microarray.

RESULTSThe fetus was found to have a 47,XX,+mar karyotype. BoBs analysis indicated that there was an amplification between 18p11.32 and 18p11.21, which was verified by the SNP-array assay as a 18.3 Mb duplication occurring at 18p11.32q11.1.

CONCLUSIONThe karyotype of the fetus was determined as 47,XX,+der18(18p11.32?18q11.1::18q11.1?18p11.32). The duplication has involved important genes including SMCHD1, LPIN2 and TGIF1, which may result in severe malformations in the fetus.

Adult ; Aneuploidy ; Chromosomes, Artificial, Bacterial ; genetics ; Chromosomes, Human, Pair 18 ; genetics ; Female ; Humans ; Karyotyping ; Microarray Analysis ; methods ; Polymorphism, Single Nucleotide ; Pregnancy ; Prenatal Diagnosis ; methods