Objective:To construct of eukaryotic expression vector of RNA interference specific for CXCR4. Methods: Genome sequences of CXCR4 gene were retrieved from Genbank and cDNA was designed coding expression of shRNA (small hairpin RNAs) for CXCR4 gene. The cDNA was synthesized and inserted into plasmid pWH1, and the recombinant pWH1-CXCR4 expression vector was identified by enzyme cutting method. Then, pWH1-CXCR4 expression vector were transfected into salivary mucoepidermoid carcinoma Mc3 cells. At last, the expression of CXCR4 in Mc3 transfected with pWH1-CXCR4 expression vector was evaluated by RT-PCR and flow cytometry analysis. Result:Compared with Mc3 cells and Mc3 cells transfected with plasmid pWH1, the Mc3 cells transfected with pWH1-CXCR4 expression vector showed a lower mRNA and protein expression of CXCR4. Conclusion:The constructed CXCR4 shRNA expression vector can block the expression of CXCR4 in salivary mucoepidermoid carcinoma cells, and it may be helpful for further study on the significance of CXCR4 on the proliferation, metastasis and experimental treatment of salivary mucoepidermoid carcinoma.

Objective To discuss the clinical value of procalcitonin (PCT) in guiding antibiotics use in children with severe diseases. Methods The clinical data of patients admitted to intensive care unit from January 2012 to July 2012 were retrospectively analyzed. The patients without antibiotics use before admission and with procalcitonin level less than 0.5 ng/ml on admission were selected. The body temperature, infection indicators and prognosis were compared between patients with and without antibiotics use during hospitalization. Results There was no difference in body temperature, PCT, C-reactive pro-tein (CRP), erythrocyte sedimentation rate (ESR) and white blood cell count (WBC) on admission between patients with and without antibiotics use during hospitalization. The PCT level was increased signiifcantly (P<0.05) on the day of starting the an-tibiotics when compared with that on admission in 60 patients while there was no change in the levels of WBC and CRP. Com-pared with the day of starting the antibiotics, body temperature declined (P<0.05) and PCT level in 56 patients reexamined was decreased (P<0.05) at 3 days after antibiotics use. Two hundred and eleven patients (98.14%) had favorable prognosis. Conclu-sions Monitoring PCT can guiding the clinical use of antibiotics.

Objective:To identify metastasis-associated genes in salivary gland adenoid cystic carcinoma(ACC).Methods:Salivary gland adenoid cytic carcinoma cell line ACC-2 and its highly metastatic ACC-M cells were used to screen the metastasis-related genes in ACC by microarray technology.Two fluorescent cDNA probes labeled with Cy3 and Cy5 dyes,were prepared from the mRNA samples of ACC-2 and ACC-M cells by reverse transcription method.The two color probes were then mixed and hybridized on the cDNA chip constructed by double dots of 1152 human genes,and scanned at two wave lengths.Differentialy expressed genes of the two cell lines were analyzed using computer.Then seven of the differently expressed genes were further validated by RT-PCR technique.Results:Of the 1,152 known genes and expressed sequence tags,26 showed significantly different expression level(minimum 2 fold) between the two cell lines.Among the 26 genes,19 were up regulated(with ratio more than 2) and 7 were down(with ratio less than 1/2).The results of RT-PCR analysis for 7 differently expressed genes were coincident with those of microarray assay.Conclusions:Down regulation of LIFR,LCP1,DPEP1 and ABLIM1,and up regulation of DCC,MMP1 and CNTN2 may be related to the highly metastatic potential of ACC-M cell line.

Objective To observe the change of lipid metabolism during adaptive heart failure.Method The transverse aorta constriction mouse models established by minimal invasive surgery technique were evaluated for early small changes of body composition by EchoMRITM whole body composition analysis,and the different gene expression was analyzed by whole genome expression microarray.Results Three weeks after the surgery,the fat composition,in the mouse models were (0.49 ± 0.13) g,which was significantly lower than that in the sham surgery group [(0.81-±0.14) g] (P =0.002).The expression of the natriuretic peptide type A significantly increased over time (R2 =0.939,P =0.005).In addition,the levels of fatty acid desaturase,long-chain fatty acid elongase 5,and N-acylsphingosine amidohydrolase increased,and the levels of acyl-coenzyme A dehydrogenase,muscle glycogen phosphorylase,and pyruvate dehydrogenase kinase decreased,showing increased fat mobilization and decreased carbohydrate utilization.Conclusions Surgery for transverse aorta constriction can cause complex metablic changes in heart.The activated lipid mobilization maybe lead to the low body fat composition.

Objective To investigate the relationship between coronary artery lesions and multiple risk factors in the patients with metabolic syndrome (MS) and coronary heart disease (CHD). Methods Totally 429 patients were definitely diagnosed with CHD by coronary arteriography and their BMI, BP, FBG, TG, HDL-c as well as age, gender, smoking, TC, LDL-c, UA were recorded. All patients were divided into two groups: CHD with MS, CHD. Results BMI, BP, FBG, TG, HDL-c, TC, LDL-c and UA were significantly higher in CHD with MS group than those in CHD group (P

Objective The aim of this study is production of organ specific animal model for studying reproductive toxicity in mice.Methods F1 generation was gotten by mating the Ddx4 -cre transgenic male mice with the Rosa26mT/mG transgenic female mice.F1 offspring and its parents phenotype was screened by molecular biological, histopathological and in vivo imaging technology.Results At molecular level, specific DNA fragment was only found in testis of F1 offspring; At the organ level, the expression of green fluorescent protein could only be observed in testis of F1 offspring; Testicular frozen sections and sperm fluorescence observation showed that green fluorescent protein were mainly expressed in the germ cell lineage such as secondary spermatocyte and spermatocyte and spermatozoon.Conclusions The production of the mice with specific germ cell expressed green fluorescent protein by Cre/loxP recombination system were built successfully.

siRNA drug research and development is becoming one of the main objectives in the future. However, due to the in-stability of siRNA and the complex environment in vivo, the safe and effective delivery of siRNA is limited in vivo. Thus, special vectors are used to assist siRNA to express biological effects. This paper reviews the advances in non-viral vector for delivery of siRNA in vivo.

Objective To confirm that rat bone marrow mesenchymal stem cells (MSC) transfected with nerve growth factor (NGF) gene in the bladder tissue of diabetic rats bladder tissues can survive and stably express NGF. Methods A diabetic rat model was constructed. The BrdU-labelled MSC transfected with NGF gene were transplanted into the diabetic rats bladder tissues. BrdUlabelled immunohistochemistry was used to observe the growth of MSC transfected with NGF gene in the diabetic rats bladder tissues. The expression of NGF mRNA and protein were checked by RT-PCR and ELISA. Results A diabetic rat model was successfully built by a single intraperitoneal injectionof STZ. The blood glucose was still high after 8 weeks. NGF gene modified MSC could be detected in the bladder of diabetic rats by BrdU-labelled immunohistochemistry. The concentration of NGF in the control group, disease group and treatment group were ( 114 ± 3), ( 70 ± 2), ( 110 ± 2) pg/ml by ELISA and mRNA quantity by RT-PCR were 0. 183±0. 004, 0. 032±0. 139, 0. 130±0. 165, respectively. Compared with the control group, the expression of NGF gene was decreased (P＜0. 05) in the incidence group. The expression of NGF gene was increased (P＜0. 05) in the treatment group compared with the disease group. Conclusions The NGF gene-modified MSC could survive in diabetic rats bladder tissues. The NGF gene in MSC could stably express in diabetic rats bladder tissues.

Objective To discuss the signiifcance of serum albumin level in assessing severity, progress and prognosis of sepsis in children. Methods The clinical data of 212 patients diagnosed with sepsis admitted to PICU from February 2010 to July 2010 were retrospectively analyzed, and 52 patients had severe sepsis and 31 patients had septic shock. Meanwhile, 110 non-sepsis patients were selected as controls. The relationships of hypoalbuminemia with pediatric critical illness score (PCIS), pediatric risk of mortality III (PRISM III) and prognosis were evaluated, and the change of albumin level in patients with dif-ferent severity of sepsis was observed. Relative factors analysis of albumin level ≤25 g/L was performed. Results As the serum albumin level was decreased, the PCIS was signiifcantly decreased while the PRISM III was increased (P<0.01). The se-rum albumin level was signiifcantly different among children with septic shock, severe sepsis and sepsis and controls (F=13.938, P=0.000). The results of relative factors analysis showed that sepsis children with an albumin level≤25 g/L had more organ failures, higher mortality, longer hospital and PICU stay and more likelihood for ventilator support (P<0.01). Lower albumin levels were accompanied with lower rates of recovery and improvement but higher mortality (rs=-0.161, P=0.000). Conclusions Hypoalbuminemia can be used as indirect indicator for severity of infection. The albumin level≤25 g/L indicated the severity of illness and prognosis in children with sepsis.

Objective To explore differential expression of protiens between patients with esophageal cancer and healthy individuals, and to screen out the tumor biomarkers to construct a dignostic model. Methods From January to August, 2008, the clinical data of 127 patients with esophageal cancer (esophageal cancer group) who had been admitted to the First Affiliated Hospital of Xinjiang Medical University and 63 healthy individuals (control group) were retrospectively analyzed. The serum proteomic profiles of the esophageal cancer pateints and healthy individuals were deteced by weak cation exchange and hydrophobic surface ProteinChip using the surface enhanced laser desorption/ionization-time of flight-mass spectrometry (SELDT-TOF-MS) technique.Serum differentially expressed markers of esophageal cancer were screened out to establish the diagnostic model for esophageal cancer. All data were analyzed using rank sum test. Results Six proteins were high-expressed in the esophageal cancer group, and the mass-to-charge ratios were 4488, 5495, 15964, 3948, 8154, 8166. Four were low-expressed in esophageal cancer group, and the mass-to-charge ratios were 8789, 6682, 8714, 6650. A diagnostic model consisting six proteins was established. A total of 124 patients were correctly diagnosed and three were misdiagnosed in the esophageal cancer group, and 60 were correctly diagnosed and three were misdiagnosed in the control group. The accuracy, sensitivity and specificity of the diagnostic model were 96.8% ( 184/190),97.6% ( 124/127 ) and 95.2% (60/63). Conclusions The diagnostic model established based on the tumor markers screened out by the SELDT-TOF-MS is highly sensitive and specific.