Objective To study the effect of lentivirus-mediated CCL5-RNAi on the biological behaviors of human breast cancer cells. Methods CCL5-specific siRNA gene was synthesized and cloned into the recombinant lentiviral vector, pGCSIL-GFP. Human high-metastatic breast cancer cells, MDA-MB-231, were infected by CCL5-siRNA recombinant lentivirus, which was set as KD group. Cells infected with CCL5-NC was as NC group, and cells cultured was as CON group. The expression of CCL5 mRNA and protein in MDA-MB-231 cells was detected by RT-PCR and western blot, respectively. Cell growth suppression and cell cycle was observed by MTT assay and fluorescence activated cell sorting (FACS). Colony formation and migration ability were determined by colony-rorming assay and Boyden chamber method. Results After infection of CCL5-siRNA recombinant lentivirus, the expression level of CCL5 mRNA and protein in MDA-MB-231 cells as well as the colony formation and migration ability decreased significantly, but cell's proliferation was not affected obviously. Compared with MDA-MB-231 (0.88± 0.15) and MDA-MB-231/CCL5-NC (1.00±0.07) cells, the expression of CCL5 mRNA in MDA-MB-231/ CCL5-siRNA decreased to 0.18±0.03, P<0.01. Compared with MDA-MB-231/CCL5-NC (1.82±0.18) cells, the expression of CCL5 protein in MDA-MB-231/CCL5-siRNA decreased to 0.33±0.13, P <0.01. Colony-forming assay and Boyden chamber method showed that the colony formation and migration ability of MDA-MB-231/CCL5-siRNA decreased markedly (P<0.05). The clone count in KD group was (0.33± 0.10), which was a significant decrease from (0.97±0.09) (NC group) and (1.04±0.07) (CON group), P<0.05. The number of cells that migrated through the chamber membrane of KD group (38± 15) was less than that of NC group (77±11, P <0.05) and CON group (69±9, P <0.05). However, MTT assay and FACS revealed that the proliferation of MDA-MB-231/CCL5-siRNA was not different from MDA-MB-231/CCL5-NC and MDA-MB-231 (P>0.05), the proliferation index (PI) of group KD, NC and CON were (0.48±0.02), (0.44±0.05) and (0.47±0.02) respectively. The difference was not statistically significant by multiple comparison (P>0.05). Conclusion CCL5-specific siRNA can specifically suppress the colony formation and migration of human high-matastatic breast cancer cells.

The bone is a common site of metastasis for gastric cancer. High-risk factors of metastatic gastric cancer include young age, poorly differentiated adenocarcinoma, Borrmann type III tumors, depth of invasion at Se and Pm levels, positive lymph nodes, and gastric cancer with concomitant body of stomach cancer. More than half of the transfer pathway belongs to the non-portal system. Cases of gastric cancer with bone metastasis but without liver metastasis accounted for 69%of the total gastric cancer cases, whereas the incidence rate of bone metastasis with lymph node metastasis (≥3 cm away from the primary lesion) was 27%. In addition to the tumor markers human chorionic gonadotropin and carcinoembryonic antigen, radionuclide scan of the bone metastasis provides diagnostic pathways and reliable basis for treatment. Chemoradiation and surgery as symptomatic treatments are alternative therapies for stomach cancer. Bone metastasis confers unfavorable prognosis, and the survival time is often less than six months.

Objective To investigate the incidence of metabolic syndrome among obese children in clinics. Methods One hundred and thirteen obese children aged 7 to 14 years were selected from clinics of nutrition(case group),and another 366 healthy students aged 7 to 14 years were served as controls.Height,body weight,waist circumference,hip circumference,blood pressure and liver ultrasound were measured,related biochemical parameters such as fasting blood glucose,fasting insulin,serum total cholesterol,triglyceride(TG),hiSh density lipoprotein and low density lipoprotein were detected,and the incidences of metabolic syndrome were obtained in two groups.Insulin resistance(IR)was evaluated by homeostasis model assessment(HOMA). Results There was no significant difference in age and gender between case group and control group(P>0.05).Body weight,body mass index(BMI),waist circumference,hip circumference,waist to hip ratio,systolic blood pressure,diastolic blood pressure,fasting blood glucose,fasting insulin,HOMA index and TG in case group were significantly higher than those in control group(P<0.01).The 75th percentile of HOMA index in control group was 3.28,and IR subgroup and non-IR subgroup were divided according to this cutpoint.In case group,body weight,BMI,waist circumference and TG in non-IR subgroup were significantly higher than those in IR subgroup (P< 0.05).Metabolic syndrome occurred in 51 cases(45.1%) in case group.The incidence of metabolic syndrome was higher in IR subgroup than that in non-IR group(50.0% vs 21.1%)(P<0.05). Conclusion The prevalence of metabolic syndrome is higher in overweight and obese children.IR has a close relationship with metabolic syndrome.

Objective To investigate the antagonist effects of sodium salicylate(NaSA) on the cisplatin(CDDP) induced hearing impairment in guinea pigs.Methods 60 guinea pigs were randomly divided into four groups: ①CDDP +NaSA(50 mg/kg) group,②CDDP +NaSA(100 mg/kg) group,③CDDP +NaSA(150 mg/kg) group and ④CDDP +NS(normal saline) control group.Auditory brainstem response(ABR) and distortion product otoacoustic emission(DPOAE) were used to evaluate their effects on hearing threshold and DPOAE amplitudes.Results The ABR responses of groupsⅠ,Ⅱ and Ⅲ were significantly lower than in the control group(P0.05).The ABR responses for group Ⅰ were significantly higher than group Ⅲ(P

Objective To investigate the correlation between 6 common triggers and ischemic stroke.Methods The demographic and clinical data of the consecutive inpatients with acute ischemic stroke were collected.A questionnaire survey of the triggers of the disease was conducted within 4 d after admission.A case cross-over study was used to compare the exposure of the 6 potential triggers (overeating,anger,negative emotion,heavy physical activity,sudden posture changes as response to a startling event,drinking coffee) at 2 h before onset (dangerous period) and at 1 d before onset during the same period (control period),and exposures to potential triggers in patients according to gender,age and etiological subtypes were further analyzed.Results A total of 369 patients were enrolled.They aged 24-93 years old (mean 61.75 ±13.57),220 patients were male (59.6％) and 149 were female (40.4％).A total of 91 patients (24.7％) exposed to at least one of the triggers at 2 h before onset (odds ratio [OR] 6.1,95％ confidence interval [CI] 3.7-9.9);OR for exposure to the sudden posture change in response to a startling event was 12.0 (95％ CI 2.4-59.3),for heavy physical activity 10.7 (95％ CI 4.2-27.6),for anger 8.0 (95％ CI 2.3-27.5),and for negative emotion 4.9 (95％ CI 2.3-10.3).There was no exposure to drinking coffee.There were no significant differences in the exposure to various triggers among the different gender,age,and etiological subtypes.Conclusions Sudden posture changes as response to a startling event,heavy physical activity,anger,and negative emotion are the triggers for ischemic stroke,attention should be paid to the influence of triggers in the prevention of ischemic stroke.

BACKGROUND:Major abdominal vascular injury is a seldom serious complication during lumbar discectomy. Once occurring, it wil cause critical condition, and incorrect rescue and operation wil lead to patient’s death or functional disorder of the limbs. ＜br> OBJECTIVE:To explore the cause, diagnosis, prevention and treatment action for major abdominal vascular injury caused by lumbar discectomy. ＜br> METHODS:One case of common iliac artery injury caused by lumbar discectomy was reviewed in the aspects of diagnosis, prevention and treatment action. ＜br> RESULTS AND CONCLUSION:The case suffered from common iliac artery injury caused by lumbar discectomy of right L 4-5 vertebrae, and achieved successful rescue by timely diagnosis, surgical intervention and artificial blood vessel transplantation. Injury of the vessels is relevant with many factors, including surgical operation, anatomy, pathology, etc. Unexplained intervertebral hemorrhage accompanied with sudden drop of blood pressure during surgery hints the possible occurrence of major vascular injury. Accurate diagnosis and instant surgical intervention is the key to rescue the patient’s life.

Coronary Vessel Anomalies ; therapy ; Coronary Vessels ; Humans ; Percutaneous Coronary Intervention

Objective To study the effects of modified Danggui Beimu Kushen Pills on the expressions of TLR2, TLR4, TLR6, TRAF6 and MyD88 in tumor tissues on MFC gastric cancer bearing mice; To discuss relevant mechanism of action. Methods MFC gastric cancer bearing mice were employed to perform anti-tumor experiment in vivo in this study. A total of eligible 48 mice were randomly divided into model group, DDP positive control group, modified Danggui Beimu Kushen Pills high-dose and low-dose groups, modified Danggui Beimu Kushen Pills high-dose and low-dose combined with DDP groups. The treatment was conducted once a day, and lasted for 14 continuous days. After the last administration of gavage orally treatment, all mice were anaesthetized and killed by cervical dislocation method to obtain tumor tissue completely for further HE staining measure and detection of TLR2, TLR4, TLR6, TRAF6 and MyD88 in tumor tissue with the method of RT-qPCR and immunohistochemistry. Meanwhile, the tumor growth was observed and the general conditions of mice were recorded. Results The model group was rich in tumor cells; the sizes of cells were different; the volume was large; the nucleus was deeply stained and the heterotypic shape was obvious, and the small focal necrosis was seen. The number of tumor cells in each administration group was reduced; the arrangement was loose; the cell volume was significantly reduced, and the nuclear pyknosis was reduced. Cell necrosis significantly increased; the number of interstitial blood vessels decreased; collagen fibers increased, especially in modified Danggui Beimu Kushen Pills high-dose and low-dose combined with DDP groups. Compared with the model group, the expressions of TLR2, TLR4, TLR6, TRAF6 and MyD88 mRNA and protein decreased in each administration group. TLR2, TLR4, TLR6, TRAF6 and MyD88 were lighter and weakly positive expressed in modified Danggui Beimu Kushen Pillshigh-dose and low-dose combined with DDP groups, the protein changes were more obvious Compared with DDP positive control group, modified Danggui Beimu Kushen Pills high-dose and low-dose groups. Conclusion Modified Danggui Beimu Kushen Pills can down-regulate TLR2, TLR4, TLR6, TRAF6 and MyD88 expression of tumor tissue in MFC gastric cancer bearing mice at both mRNA and protein levels to play anti-tumor pharmacology action.

Objective To establish Ad-LIF-OSM transgenic feeder cells for the expansion of CD34+ hematopoietic stem/progenitor cell and tentatively study its effect in expansion and differentiation of cord blood hematopoietic stem cell(HSC) in vitro.Methods In the foundation of pAdTrack-CMV-LIF-polyA+promoterΔ,the OSM gene was inserted to the vector plasmid.Then we structure the transfer plasmid pAdTrack-CMV-LIF-polyA+promoterΔ-OSM.The transfer vector and backbone vector were further cotransfected for homologous recombination. The result pAdEasy-1-pAdTrack-CMV-LIF-polyA + promoterΔ-OSM homologous recombination plasmid were transfected into the human embryonic kidney 293 (QBI-293A) cells,leading to formation of the recombinant adenviruses Ad-LIF-OSM which co-expressing LIF and OSM.Infect the feeder layer cells with groups of Adenovirus,detection the expressing of LIF and OSM in WI-38 cells by RT-PCR and ELISA.Compares the stem cells differentiation and proliferation of the different experimental groups in vitro by transwell and cell counting.Results The sequencing results show that the OSM genes were anastomotic in Ad-LIF-OSM.LIF and OSM gene could be detected in feeder layer cells which infected by Ad-LIF-OSM.Exogenetic LIF and OSM have special effect in culturing HSC in vitro.Conclusion The adenoviral vector co-expressing LIF and OSM (Ad-LIF-OSM) were successfully constructed.Ad-LIF-OSM transgenic feeder cells can effectively proliferate umbilical cord blood CD34+ HSPC in vitro and delay it differentiate.

OBJECTIVE To investigate the expression of cycloxygenase-2(COX-2)and matrix metalloproteinase-7(MMP-7)in laryngeal squamous cell carcinoma(LSCC)and their clinical significance. METHODS The expression of COX-2 and MMP-7 in 65 LSCC tissues and 34 adjacent non-neoplastic tissues were studied by immunohistochemical SP method. RESULTS The expression of COX-2 and MMP-7 was observed in 72.3% and 75.4% of the LSCC tissues and in 20.6% and 23.5% of the adjacent non- neoplastic tissues respectively,which had a significant difference(P