Objective To study the effects of lentiviral vector of microRNA targeting IGF1R gene on growth of liver cancer cells.Methods The complementary DNA containing both sense and antisense Oligo DNA of the targeting sequence was designed,synthesized and cloned into the pPRIME vector,named pPRIME-IGFIR-miR30-shRNA.The viruses were propagated on 293T cells.Viruses were purified by CsCI gradient according to standard techniques,and functional PFU titers were determined by plaque assay on 293 cells.The effect of pPRIME-IGFIR-miR30-shRNA on IGFIR expression of Hep3B、SMMC7721 cells was detected by RT-PCR and Western blot.The antitumor potential of PRIME-IGFIR-miR30-shRNA to Hep3B、SMMC7721 cells was evaluated by CCK-8 assay and Tunel.Results PRIME-IGFIR-miR30-shRNA was constructed successfully.Functional PFU titers of pPRIME-IGFIR-miR30-shRNA were 4.58?109PFU/ml.PRIME-IGFIR-miR30-shRNA inhibited IGFIR expression and the proliferation of Hep3B、SMMC7721 cells.Conclusions PRIME-IGFIR-miR30-shRNA expressing IGFIR-siRNA can inhibit IGFIR expression and may be used for further investigation of gene therapy of liver cancer.

Objective To investigate the effects of keratinocyte growth factor(KGF) and keratinocyte growth factor receptor(KGFR) on the malignant transformation of gestational trophoblastic disease(GTD).Methods Immunolocalization of KGF/KGFR was performed on sections prepared with the samples from 26 hydatidiform mole,18 invasive mole and 12 choriocarcinoma.The in situ hybridization was used to detect the mRNA of KGF/KGFR in the tissues of hydatidiform mole and GTD.Analysis was performed according to intensity of staining and number of positive cells.Results It was revealed that specific staining for mRNA and protein of KGF/KGFR existed in hydatidiform mole and gestational trophoblastic tumor(GTT).The mRNA and protein of KGF/KGFR were allocated in cytoplasm of syncytiotrophoblasts and cytotrophoblasts of malignant hydatidiform mole,and the KGF/KGFR protein was also expressed in benign tissue,while the expression of KGFR in malignant hydatidiform mole was significantly higher than that in benign tissue(?2=12.775,P

Objective To observe the histopathological changes in the kidneys with stress reaction under +Gx loads in rhesus monkeys. Methods Nine male Rhesus macaques monkeys were randomly divided into four groups: control group animals were exposed to +1Gx/300s overload, and experimental groups 1, 2 and 3 animals were exposed to +15Gx/200s, +21Gx/165s and +21Gx/140s (3 animals in each group), respectively. Renal tissue was fixed with 10% buffered formaldehyde and paraffin sectioned, and histopathological changes were observed with microscope. Results In the experimental groups, glomerular congestion, and dilation of interstitial vessels with erythrocytic extravasation were found in both kidneys. Obvious cloudy swelling of the epithelial cells was also noted in the renal tubules. The degree of renal damage was significantly correlated with the increase in +Gx loads. There was no obvious change in the control group. Conclusion Overload gravity can induce significant injury to the kidney in the monkeys. There is a significant correlation of the degree of renal damage with the level of +Gx load.

Neuromuscular diseases is one of the difficulties in neurology teaching and clinical practice. We chose the typical cases of Guillain-Barre syndrome and progressive muscular dystrophy as case based study, and evaluated the effect of teaching. The result shows that application of case base study can help medical students to enhance their learning interests and cultivate their good clinical thinking and then to meet the requirements of the teaching syllabus. So it is worth improving and promoting in clinical practice teaching.

Objective To explore the acute effects of nickel in air PM2.5 on the cardiovascular system of rats.Methods The air PM2.5 of nickel-contaminated area and control area were collected and determined for some major metal elements.42 male Wistar rats were randomly divided into 10 groups,6 rats in each,the rats were instilled through trachea with saline,PM2.5 suspension of nickel-contaminated area and the control area respectively,at different doses(low,moderate and high dose).24 hours later,all the rats were killed for the determination of sVCAM-1 in the blood and MCP-1 in the myocardial tissue.Results The concentration of nickel in the air PM2.5 of nickel-contaminated area was found to be about 50 times higher than that in the control area.A good dose-response relationship was found in MCP-1 determination of the heart tissue in nickel-contaminated groups,and there were significant differences between nickel-contaminated area group and other groups(P﹤0.05).As for the sVCAM-1 in the serum,it was significant higher in the nickel-contaminated group and control group than the saline control group(P﹤0.05).Among the groups of nickel-contaminated area and control area,significant differences were found between the moderate and high dose groups.Conclusion The air PM2.5 from nickel-contaminated area has an obvious effect on the MCP-1 and sVCAM-1 of rats,the nickel in the air PM2.5 is likely the main contributing factor.

Epithelial mesenchymal transition (EMT) is a process of normal cell physiological development, in which epithelial cells transform into mesenchyme cells through a specific program. EMT plays a key role in inflammatory reaction, cell development, tumor invasion, and metastasis and has an interrelation with prostate cancer stem cells. Recent researches show the involvement of EMT in the development and metastasis of prostate cancer. This article reviews the specific roles and action mechanisms of EMT in the progression of prostate cancer.
Biomedical Research ; Cell Differentiation ; Disease Progression ; Epithelial Cells ; physiology ; Epithelial-Mesenchymal Transition ; physiology ; Humans ; Male ; Mesenchymal Stromal Cells ; Neoplastic Stem Cells ; physiology ; Prostatic Neoplasms ; pathology

Objective To construct lentiviral vector miR30-CD133 targeting CD133 gene and investigate its effects on SMMC7721 cells. Methods Thespecific sequence of DNA which containing both sense and antisense Oligo DNA of the targeting sequence were designed, synthesized and cloned into the pPRIME vector. pPRIME-miR30-CD133, psPAX2 and pMD2G were co-transfected on 293T cells to get the lentivirus the transfection efficiency was investigated by confocal laser scanning microscope. The expression of miR30-CD133 on CD133 were detected by qRT-PCR and Western blott. The proliferation and apoptosis effect of miR30-CD133 was respectively evaluated by CCK-8 assay and AnnexinV/PI. Results Functional PFU titers of PRIME-miR30-CD133 were 6.58 × 109 PFU/mL. The expression of CD133 mRNA in sh CD133 group (0.18 ± 0.04) was less than Blank group and shNC group (P<0.05). The expression of CD133 protein in sh CD133 group was less than Blank group (10%) and shNC group (8%) (P < 0.05). Cells proliferation activity were significantly inhibited in shCD133 group compared with control group. Apoptosis rate were significantly increased in shCD133 group (41.3%) compared with Blank group (25.3%)and shNC group (24.3%). Conclusion The lentivirus miR30 vector targeting CD133 gene was successfully constructed, which will be a basis to the further CD133 gene studies.

Background Researches have demonstrated that ocular hypertension induces the ischemia-reperfusion of retina and further leads to the degeneration of retinal ganglion cells,but its mechanism is beyond understanding.Objective The present study aims to observe the effects of static pressure on the morphology,proliferation activity and viability of cultured retinal microvascular endothelial cells (RMECs) and evaluate the expression of ET-1 and NO in these cells under variant static pressure.Methods RMECs were isolated from 30 healthy Wistar rats and cultured using explant culture method by Ⅷ factor antibody and PECAM-1 antibody.The static pressure of 1.33kPa,2.67kPa,5.33kPa and 10.67kPa was used in culture bottle respectively.The RMECs without static pressure were used as normal control group.The morphology of RMECs under the different static pressure was observed by inverted phase contrast microscopy,and the number of RMECs was counted using the counting plate.Cellular viability was studied by trypan blue staining.The changes of ET-1 and NO_2~-/NO_3~-,two metabolic products of NO,in the medium were detected by radioimmunoassay and Griess's nitrate reductase method.The expression of ET-1,eNOS and iNOS mRNA in RMECs was analyzed by semi-quantitative RT-PCR 24 hours after treatment of variant static pressure.Results Cultured RMECs sticked well at 24 hours and reached to confluence at 48 hours and showed the red fluorescence for Ⅷ factor antibody and PECAM-1 antibody.Enlargement of nuclei,extenders of cell bodies and suspension of RMECs in medium were observed.The number of RMECs was gradually increased.The cell viability was reduced with the raise of static pressure among these four groups(F=12.205,P＜0.01;F=11.180,P＜0.01).The static pressure increased the content of ET-1 released by RMECs in 2.67kPa,5.33kPa and 10.67kPa of static pressure groups,and concentrations of NO_2~-/NO_3~- in the medium showed a significant increase in 5.33kPa and 10.67kPa of static pressure groups compared with normal and 1.33kPa of static pressure groups(P＜0.01).The expressions of ET-1 mRNA,eNOS mRNA and iNOS mRNA were considerably enhanced in 5.33kPa and 10.67kPa of static pressure groups compared with normal control group(P＜0.01).Conclusion Raised static pressure causes the alteration of RMGCs structure and morphology.Static pressure could upregulate the expressions of ET-1,eNOS and iNOS mRNA in RMECs and increase the release of ET-1 and NO.This pathway might be one of pathologic mechanisms of retinal injury induced by high intraocular pressure.

0.05).Conclusion The SIS method not only can determine multiple index in Chinese materia medica(CMM)simultaneously,but also be simple and accurate.It will be a good quality evaluation pattern for CMM.