Objective: To investigate the effects LASS2 gene on the metastasis of human hepatocellular carcinoma cell line, HCCL3. Methods: The expression level of LASS2 mRNA in HCCLM3 cells was analyzed by northern blotting in high metastatic HCCLM3 cells and in low metastatic MHCC97-L cells. The eukaryotic expression vector pCMV-HA2-LASS2 was constructed and transfected into HCCLM3 cells. The stable clone with LASS2 overexpression was selected by G418 screening. The metastatic ability of HCCLM3 cells such as migration and invasion was observed in vitro after LASS2 overexpression. The synthesis, secretion, and activation of matrix metaloproteinase-2 (MMP-2) were examined by western blotting, zymography, and in situ zymography. Results: Northern blot analysis showed that the level of LASS2 mRNA decreased in HCCLM3 cells compared with MHCC97-L cells. LASS2 overexpression inhibited the ability of migration and invasion of HCCLM3 cells (P <0. 001 ). The synthesis of MMP-2 was not different but the secretion and activation of MMP-2 were down-regulated in HCCLM3 cells afterLASS2 overexpression. Conclusion: Overexpression of LASS2 down-regulated the secretion and activation of MMP-2 in HCCLM3 cells and inhibited the cell ability of migration and invasion indicating that LASS2 gene had inhibitory effects on the metastasis of HCCLM3 cells.

Alkaline Phosphatase ; blood ; Biomarkers ; blood ; Child, Preschool ; Female ; Humans ; Infant ; Isoenzymes ; blood ; Phosphorus Metabolism Disorders ; blood ; diagnosis ; Risk Factors

Acupuncture Points ; Adult ; Electroacupuncture ; Fatty Liver ; therapy ; Female ; Humans ; Male ; Middle Aged ; Non-alcoholic Fatty Liver Disease

Objective To establish stable Graves disease (GD) mice models with immunization and electroporation (EP).Methods Fifty mice were divided into 3 groups by random number table method:experimental group (n =30),control group (n =10),blank group (n =10).Recombinant plasmid pcDNA3.1/hTSHR268 was constructed and injected to bilateral gastrocnemius in experimental group mice on the 1st,4th,7th and 10th week.The same volume of normal saline was injected in the control group and blank group at the same time.Both experimental group and control group were subjected to EP at the same time and the same location to enhance immunization.Serum T4 was tested with radioimmunoassay.TRAb N-terminal (TRAb N) and TRAb C-terminal (TRAb C) antibodies were tested with ELISA.Whole body 99TcmO4-imaging was performed and then thyroid morphology and pathology were investigated.Data were analyzed by one-way analysis of variance and the least significant difference (LSD) t test.Results GD BALB/c mice models were built successfully (80％,24/30).Serum T4 increased from (16.06±5.16) nmol/L at the basic level to(95.04±68.92) nmol/L on the 12th week(F=18.906,t=-5.598,P＜0.05).Serum TRAb N antibody increased from (0.006±0.002) U/L at the basic level to (0.251±0.110) U/L on the 12th week(F=47.491,t=-10.869,P＜0.05).Serum TRAb C antibody increased from (11.176±2.635)×103 arbitrary unit (AU)/L at the basic level to (46.395±22.001)× 103 AU/L on the 12th week(F=14.642,t =-7.787,P＜0.05).On the 18th week serum T4,TRAb N and TRAb C decreased to (36.64±23.68) nmol/L,(0.094±0.053) U/L and (24.456±6.725)× 103 AU/L respectively,which were still higher than those preimmune levels(t=-4.161,-8.085,-9.008,all P＜0.05).There were no significant change of T4,TRAb N and TRAb C in the control group and blank group.After 4 times of immunization,the 99TcmO4-uptake by thyroids in immunized mice increased.The thyroid glands of immunized mice showed enlargement.Microscope examination showed that there were lymphocytes infiltration,colloid decrease and epithelial cell proliferation in thyroids of immunized mice.Conclusion GD mice models were successfully established by injecting recombinant plasmid pcDNA3.1/hTSHR268 and EP.

BACKGROUND:Human umbilical cord blood contains a large number of hemopoietic stem cels and mesenchymal stem cels. Neonate umbilical cord blood-derived stem cels show milder immunological rejection and lower immunogenicity than peripheral blood- and bone marrow-derived mesenchymal stem cels. OBJECTIVE:To investigate the feasibility of umbilical cord blood-derived CD133 cels for treatment of gestational diabetes, so as to provide novel methods for clinical treatment of gestational diabetes. METHODS:Mouse models of gestational diabetes were established by high-fat and high-sugar diet feeding. Umbilical cord blood-derived CD133 cels were sorted by flow cytometry sorting technique and then transplanted into mouse models of gestational diabetesvia the tail vein. At 7 days after cel transplantation, serum levels of fasting blood glucose, fasting insulin, total cholesterol and triacylglycerol were measured and mouse pancreatic tissue injury and repair was observed by hematoxylin-esoin staining. Mouse insulin resistance index and pancreatic islet function were analyzed using the homeostatic model assessment. RESULTS AND CONCLUSION:Umbilical cord blood-derived CD133 cels could be isolated from umbilical cord blood monocytes using flow cytometry sorting technique, with cel purity of (90.24±2.56)%. At 7 days after cel transplantation, serum levels of fasting blood glucose, fasting insulin, total cholesterol and triacylglycerol were significantly decreased, insulin resistance index was significantly decreased, and pancreatic islet function was significantly improved in mouse models of gestational diabetes (alP < 0.05). In addition, atrophy of pancreatic tissue and infiltration of inflammatory cels were reduced. These results show that umbilical cord blood-derived CD133 cel transplantation can improve the disease condition of gestational diabetes by repairing injured pancreatic tissue, decreasing insulin resistance index and improving pancreatic islet function.

Objective To probe the reasons of failure of caged cervical intervertebral fusion and define the indications, operative techniques and short term results of the revision surgery. Methods Twenty-seven cases of caged cervical fusion were revised. The indication for fusion was cervical disc herniation in four cases and cervical spondylotic mylopathy in 23 cases. Of the 27 cases, there were 8 single levels, 15 double levels, 4 three levels. The intervals from revision to primary fusion were 2-25 months, 10.3 months in average. Patients presented discomfort (22 cases) and local pain of neck and shoulder (9 cases) or paralysis (19 cases) preoperatively. JOA grade was mean 11.6 points before revision. Radiographic examination showed the mean lordosis loss was 7.1 mm, the mean interbody height loss was 3.9 mm. 24 patients presented kyphosis. Occluded neighboring cages were found in multilevel cases. Nonunion was founded in 6 cases, unstable in 9 cases, compression still existed in 19 cases. Removal of cages and then decompression, bone grafting and anterior plating fixation were performed in 23 cases (29 cages). 16 patients underwent further posterior fixation. Posterior revisions underwent in another 4 cases (9 cages), including laminectomy and reshaping the lordosis and lateral mass screw plating fixation. Results All the cases were followed-up from 4 to 26 months, 11.7 months in average. The local symptoms released in 81% patients. Nerural deficit symptoms improved in 58% patients. Mean JOA grade was 14.2 points after revision. In 23 cases of anterior approach revisions, 17 cases obtained bone healing within 3 months, while 6 cases got delayed union. The kyphosis was corrected, and the intervertebral height increased 3 mm in average. There were no nerural impaction or failed internal fixation. Conclusion Deficient decompression, sinking of the cage into the cancellous bone of the vertebral body and subsequent kyphosis and bone graft nonunion consist of the main causes for revision surgery. The indications of revision are aggressive neurologic symptom, unstable or deformity of the cervical spine. Cage removal, completely decompression, bone grafting and secure fixation are major steps of revision procedure. In case of more than 3 fusion levels, further posterior fixation is advised.

Objective To investigate the level of serological interleukin 35 (IL-35) in breast cancer patients and its clinical significance.Methods From Jan 2015 to July 2016,serum of 55 patients with breast cancer were collected from a tertiary hospital of Nanjing.Their pathological stages were also analyzed,including 12 cases of stage Ⅰ,15 cases of stage Ⅱ,12 cases of stage Ⅲ,and 15 cases of stage Ⅳ.Meanwhile,serum from 54 healthy individuals also selected.The level of serological IL-35 was determined by ELISA,and the IL-35 level,pathological stage and tumor metastasis were analyzed by correlation a nalysis.Results The serum IL-35 in patients with breast cancer was significantly elevated than health individuals.With the increase in pathological stage,IL-35 level in breast cancer patients with stage Ⅲ and Ⅳ was significantly higher compared to patients with stage Ⅰ breast cancer (P＜0.05).Further,patients with breast cancer metastasis had increased level of IL-35,compared to breast cancer patients without tumor metastasis (P＜ 0.05).Finally,Spearman correlation analysis indicated that the serum level of IL-35 in patients with breast cancer was positlvely correlated with the pathological stages (r=0.390,P=0.004) and tumor metastasis (r=0.361,P=0.008) of the patients,respectively.Conclusion High level of IL-35 was detected in serum from breast cancer patients,and its expression was highly correlated with pathological stage and tumor metastasis.Detection of IL-35 could be applied as a valuable potential biomarker for the diagnosis of breast cancer.

0.05).Conclusion CO2-independent culture cell system is parallel to CO2-dependent culture cell system in their effects on cell proliferation and gene expression.

To investigate the changes in CAP and CM recorded from the cochlear round window when glutamate solution in different concentrations(10mmol/L, 5mmol/L, 1mmol/L) was instilled into whole cochlea perilymph. The results demonstrated that there was a dependent correlation between CAP threshold shift and glutamate concentration. CAP threshold shift and amplitude could be markedly influenced by glutamate instillation. However, amplitude of CM was not obviously affected, and the I/O function of CM amplitude always maintained a nonlinear characteristic. The results suggest that glutamate has selective effects between inner hair cells and outer hair cells. High concentration glutamate might cause an increase in intracellular calcium, even as high as to induce toxicity. It could be induced by glutamate when it acts on IHC′s presynaptic autoreceptor in positive feedback manner, or on the afferent neurons, so that the synapsis is directly damaged by glutamate concentration overload in synaptic cleft.

Objective To explore how the freshmen's self-efficiency influence the acute stress psychological reaction. Methods Self-efficiency, coping styles, personality were assessed by relevant questionnaires, and acute stress psychological reaction was measured by means of designing a experiment in 683 freshmen of military colleges.Results ①Freshmen with low self-efficiency had higher scores in Si,Mas,Dy,Nc(t=8.05,7.49,6.36,3.93,P<0.01)and lower scores in Do,Re,Pc(t=5.12,2.11,3.90,P<0.05 or P<0.01)than freshmen with high one.Among all factors mentioned above, Si,Dy,Pc got into regression equation. ② Freshmen with low self-efficiency had lower stress score than ones with higher self-efficiency(t=3.92,P<0.01).Freshmen's self-efficiency was positively correlated to total scale score and 3 subscale scores of the acute psychological stress reaction questionnaire(r=0.187～0.346,P<0.05,P<0.01).Conclusion Personality,coping style exerted influence on the freshmen's self-efficiency which is a mediator between stressors and acute psychological stress reaction.