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.

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 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.

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 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.

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.

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 investigate the clinical value of the transluminal radiofrequency catheter ablation (RFCA) for malignant esophageal obstruction.Methods The retrospective cross-sectional descriptive study was conducted.The clinicopathological data of 52 patients with malignant esophageal obstruction who underwent transluminal RFCA at the Affiliated Hospital of Shandong Academy of Medical Science between March 2013 and March 2016 were collected.Patients received the bipolar radiofrequency ablation (RFA) under dualchannel endoscopy and X-ray.Observation indicators:(1) intra-and post-operative situations:operation situations,operation time,time of RFA,postoperative complications and duration of postoperative hospital stay,(2) follow-up.Follow-up using outpatient examination and telephone interview was performed to detect the subsequent treatment,survival of patients and recurrence of esophageal obstruction up to June 2016.Measurement data with normal distribution were represented as average (range).Results (1) Intra-and post-operative situations:52 patients underwent successful RFCA,without the occurrence of aspiration,asphyxia,hemorrhage and perforation.Esophageal obstruction was disappeared after treatment,X-ray findings showed a smooth esophagus.Average operation time and time of RFCA were respectively 58 minutes (range,20-71 minutes) and 23 minutes (range,8-42 minutes).Patients took liquid food at postoperative day 2 and normal food at postoperative day 3,without the sensations of esophageal obstruction.Of 52 patients,1 with postoperative hypotension returned to normal level through rehydration and increasing blood volume.Five patients with postoperative substernal pain were improved after 2-day symptomatic treatment.And other 46 patients didn't have postoperative complications.Average duration of postoperative hospital stay was 3 days (range,1-5 days).(2)Follow-up:52 patients were followed up for 3-24 months,with a median time of 13 months.Of 52 patients,17 underwent single intravascular interventional therapy,15 underwent intravascular interventional therapy combined with single systemic chemotherapy,14 underwent single systemic chemotherapy and other 6 didn't undergo antineoplastic therapy.During the follow-up,9 patients didn't have esophageal obstruction and 26 were complicated with esophageal obstruction again.Esophageal obstruction of 26 patients was respectively occurred at 3-8 months postoperatively,20 patients were improved after bipolar transluminal RFCA under dual-channel endoscopy and X-ray and 6 received parenteral nutrition support therapy due to extreme exhaustion.Seventeen patients died of cachexia caused by terminal malignant tumors.Conclusion Transluminal RFCA is safe and effective for malignant esophageal obstruction,with a good short-term outcome.

Objective To report the clinical,electrophysiological and genetic features in a family with Charcot-Marie-Tooth disease type 1D (CMT1 D).Methods The proband,a 53-year-old man who was found with pes cavus when he was 15 years old,presented with weakness in both lower limbs at the age of 37,aggravated and numbness in legs at the age of 50.His daughter was confirmed pes cavus in her teens and weakness in both lower limbs at the age of 18.Electrophysiology and next generation sequencing were performed in the proband.Results Electrophysiological results of the proband showed demyelinating change in motor and sensory nerves.Latency prolongation was found in bilateral waves Ⅲ,V and abnormal differentiation in bilateral waves Ⅰ of brainstem auditory evoked potential,while both interpeak latencics of Ⅲ-Ⅴ were normal.DNA analysis revealed a heterozygous 1141C ＞ T mutation in exon 1 of early growth response 2 (EGR2) gene in both of the proband and his daughter.Conclusions The onset age of Arg381Cys mutation in EGR2 gene could be at juvenile with weakness in both lower limbs.The phenotype of CMT1D is mild and progressive slowly.

Objective To detect the levels of CD4+CD25HiCD127Low regulatory T cells (Treg) in the peripheral blood and its clinical significance in patients with esophageal cancer. Methods The levels of Treg in the peripheral blood were detected by three-color flow cytometry (FCM) in 80 patients with esophageal cancer and 20 healthy controls. Among the 80 patients, 30 patients were also further studied for preoperative and postoperative comparison after operation. The clinical and pathological data of each patient were collected and analyzed for the correlation with the Treg levels. Results The level of Treg in the peripheral blood of the control group was lower than that of the esophageal cancer patients [(3.36±1.14) % and (5.70±1.96) %, respectively], with significant difference (P <0.01). The levels of Treg in the peripheral blood was higher in the patients with metastasis of lymph node (n=40) than that in the patients without metastasis of lymph node (5.96±1.36) % and (4.23±1.18) %, respectively] (n=30), with significant difference (P <0.01). The levels of Treg in the peripheral blood of the patients were negatively correlated with their TNM classification. As the TNM classification advanced, the level of the Treg in the peripheral blood increased. Conclusion The levels of Treg in the peripheral blood of the patients with esophageal cancer are significantly higher than that of the healthy subjects, which is correlated with the clinical and pathological conditions. The development of esophageal cancer may relate with suppression of immune function.