Objective: To explore the influence factors of the degenerate oligonucleotide primered PCR(DOP-PCR). Methods: Genome DNA template from the mouse single oocyte or liver tissue were used to perform DOP-PCR. DOP-PCR was carried out with templates of different origin, different gradient dilution, with or without low melting point gel purified to wipe off the small fragment that might interfere with the following analysis, and then PCR of gene FTCD and CBS were carried out to evaluate the influence of these factors on the amplification efficiency and specificity. Results: Compared with genome DNA template from mouse liver, the template from single oocyte had the same efficiency and specificity but a minor yield and different gradient dilution of DNA template had no effect on the efficiency and specificity. Furthermore, there was a higher specificity in the low melting point gel-purified DOP-PCR product than in untreated ones. Conclusion: We have got a satisfactory result and increased specificity from DOP-PCR product purified with the low melting point gel. Single oocyte of mice could be used for further investigation of special genes detection by DOP-PCR and of an optimization in the yield of the products.

Objective To investigate the effects of hedysarum polybotys saccharide (HPS) on lipid metabolism and the expression of stearoyl-CoA desaturase-1(SCD-1) gene in rats with non-alcoholic fatty liver disease (NAFLD). To discuss the interfering effects of HPS on NAFLD. Methods The SD rats were randomly divided into the blank control group and the experiment group. Rats in the experiment group were fed with lipid rich food for 8 weeks to establish model and were randomly divided into model group, drug positive group and HPS group. After 8 weeks of drug intervention, the level of ALT, AST, TC, TG, HDL-c and LDL-c were measured with automatic chemistry analyzer, and expression of SCD-1 gene was measured by semi-quantitative polymerase chain reaction.Results Compared with blank control group, serum ALT, AST and TC, TG, LDL-c of model group were higher (P<0.05,P<0.01), the level of HDL-c of model group and the expression of SCD-1 gene were lower (P<0.01). Compared with model group, HPS was useful to decrease serum ALT, AST, LDL-c, TC and TG (P<0.05,P<0.01), and increase the level of HDL-c (P<0.01) and the expression of SCD-1 gene (P<0.01).ConclusionHPS had a positive effect on regulating lipid metabolic disturbance of NAFLD rats and promoting the expression of regulatory gene SCD-1.

Objective: To investigate the direct protective effect of SiniTang (SNT) on cultured myocardial cells of neonatal rats injuried by hypoxia and reoxygenation. Methods: The contracting frequency and content of primary-cultured myocardial cells sampled from different groups (control, oxygen and glucose deprivation model, and theated groups with 8. 0, 17. 9, 40, 89. 4, 200?g/mL SNT) were determined by TV recording system and computer image analysis system at different time (hypoxia for 30min, 60min, 90min, 120min, 150min, 180min and reoxygenation for 30min, 60min). The LDH activity in culture media was measured colorimetrically. The quantity of surviving myocardial cells was measured with MTT reduction method. Results: The contraction frequency and range of cultured myocardial cells decreased during hypoxia, the myocardial cells'pulsation of the groups treated with SNT was kept longer than the oxygen and glucose deprivation model, and the myocardial cells treated with SNT of 200?g/mL pulsed again after reoxygenation. After hypoxia for 3 hours, the LDH release of oxygen and glucose deprivation model was significant higher than that of the control ( P

Objective: To clone a novel gene and explore its expression patterns in tissues and cells,so as to find its role in the process of encephalopathy in DS.Methods: On the base of our previous microarray's result together with the tissue type,we chose EST AI480014 to carry out RACE,then analyzed its expression profiles in liver,spleen,kidney,heart,brain by multi-tissues Northern blot,after that semi-quantitive RT-PCR was used to reexamine the expression profiles.Furthermore,we used ISH to find whether aim gene expressed in neuroglial cells cultured in vitro.Finally we performed semi-quantitive RT-PCR to explore whether it expressed differently between DS and normal.Results: We gained a 682 bp new cDNA fragment(DQ275636)which expressed in all the tissues examined and had no alternative splices in them.It expressed highly in brain especially in frontal lobe and hippocampus.According to the ISH result we convinced that it expressed in neuroglial cells.Using bioinformatics we mapped DQ275636 to chromosome 5q14.Conclusion: We have obtained a new gene fragment based on the(above) results.According to its expression character and tissue type,it can be suggested that this gene has a probable role in the process of encephalopathy in DS.

OBJECTIVE Study the role of estrogen receptor (ER)in the inhibition of cell viability and differentiation induced by bisphenol A (BPA)in micro mass culture of rat e mbryonic midbrain(MB) cells.METHODS Micro mass cultures of MB were prepared fro m rat e mbryonic midbrain on gestation day 13.MB cells were exposed to BPA (10 -4 ,10 -6 ,10 -8 ,10 -10 ,10 -12 mol·L -1 )for 5 d.Cell viability was assessed by neutral red uptake test.MB differentiation was detected by he matoxylin staining and i mage analysis.In order to observe the role of ER pathway in the toxicity induced by BPA,cell cultures were co-treated with ICI182780 0.1 n mol·L -1 ,ta moxifen 1 n mol·L -1 and BPA 0.1 mmol·L -1 for 5 d, the cell viability and foci differentiation were detected.Moreover,the protein expression levels of ER in normal e mbryonic brain of gestation day 18,testis tissue fro m adult rats and midbrain cells untreated with BPA were investigated by Western blot.The mRNA expression levels of ER in normal e mbryonic brain of gestation day 13 and gestation day 18,ovary and testis tissue fro m adult rats,and midbrain cells un-treated with BPA were investigated by real-ti me PCR.The mRNA expression levels of Notch1 and Hes1 in MB cells treated with BPA 0.1 mmol·L -1 were also detected by real-ti me PCR.RESULTS BPA 0.1 mmol·L -1 could inhibited MB cell viability and foci differentiation.However,this effect could not be reversed by ER antagonist.The protein and mRNA expression levels of ER in e mbryonic brain and MB cells untreated with BPA were found to be extre mly low.In addition,BPA 0.1 mmol·L -1 could inhibited the mRNA expression levels of Notch1 and Hes1 .CONCLUSION BPA could inhibited MB cell viability and foci differentiation.ER pathway might be not involved in this effect.Instead,Notch-Hes pathway might be involved for this effect.

Objective To explore the event-related potentials (ERPs) P300 is changeable or not before and after treatment. Methods 99 cases of patients with first onset of depression diagnosed by DSM-Ⅳ as case group,and 100 cases matched with patients as control group were collected. P300 of two groups were obtained before and after treatment for 6 weeks,12 weeks,24 weeks. T test was used to analysis the difference of indicators of P300 among groups; repeated measure analysis of variance was used to analysis the longitudinal changes. Results shorter latency of N2-P3 ( (P ＜ 0.01 ); and lower amplitude of N2, P3, N2-P3 (P ＜ 0. 05 ), higher amplitude of P2-tency and a upward one in N2-P3 latency in the four periods; a upward trend could also be found in P3, N2-P3 amplitude, but there were no statistical differences(P ＞ 0. 05 ). The results of paired-samples t test: P3, N2-P3 amplitude in case group were higher after treatment for 6 weeks than before, the difference was significant (P ＜ 0.01 ); no significant results were found in P300 latency or amplitude between the 62 cases of depression after treatment for 24 weeks and the 65 normal controls selected (P ＞ 0. 05 ). Conclusion P300 latencies and amplitudes tend to be partly recovered after the acute treatment in patients with depression, but after the long-term therapy not clear.

Objective To explore the effect of posterior debridement, grafting and internal fixation for treatment of non-specific lumbar intervertebral infection. Methods Clinical data of 20 patients with non-specific lumbar intervertebral infection treated in General Hospital of Ningxia Medical University from October 2013 to June 2013 were retrospectively analyzed. There were 15 males and 5 females with an average age of 41 years (range, 36-51 years). All patients suffered from single lumbar inter-vertebral infection, including 3 cases at L2/3,4 at L3/4,10 at L4/5 and 3 at L5/S1. All 20 cases underwent one-stage posterior debride-ment, autogenous bone grafting and internal fixation, tissue samples in focus were collected for bacterial culture and pathological examination. The disease controlling statues were evaluated based on laboratory results of ESR and CRP. Imaging examinations were taken to evaluate the fusion of vertebral body. Clinical effects were evaluated using the visual analog scale (VAS) and the Jap-anese Orthopaedic Association scores (JOA) score of lumbar fumction. Results All patients underwent the surgery successfully. The surgery duration time was 90-160 min, average 125 min, and the blood loss was 200-700 ml, average 360 ml. Cerebrospinal fluid leakage occurred in one case. Postoperatively, all patients experienced significant reliefof back pain, improving in the func-tion of movement, and no fever. The lower back VAS score: average (5.35 ± 1.15) points before operation , average (2.76 ± 0.34) points one week after operation, and an average score of (0.85±0.65) points by the last follow-up time. JOA lumbar function score:all patients were effective after operation, the improvement rate was excellent in 65%(13cases), good in 25%(5 cases), and pass-able in 10%(2cases). Comparing with preoperation, the excellent and good rate was 90%. All patients ESR and CRP returned to normal levels at the last follow-up. Ordinary bacterial culture was positive in 8 cases and negative in 12 cases. The pathogens iden-tified were staphylococcus aureus (6 cases), Escherichia coli (2 cases) and staphylococcus epidermidis (2cases). All incisions achieved primary healing. All patients were followed up from 6-18 months (average,12 months), and the symptom of pain relieved significantly. No recurrent infection had happened. A solid bony fusion was found in all patients at 6-14 months (average, 8.5 months) after the surgery. Conclusion Posterior debridement, grafting and internal fixation are effective treatments for non-spe-cific lumbar intervertebral infection, can reduce the time of staying in hospital, this operation is safe and reliable.

Cervical spondylosis is due to degenerative cervical disc and its stimulation or oppression of the adjacent nerves, spinal cord, spinal artery and other tissue caused by clinical symptoms. The cervical spine is an anatomical structure with activity, while the pillow has a certain plastic fixation effect on the cervical spine anatomy. Therefore, the pillow not only plays a health role in the cervical spine, but also plays an important role in restoring the normal physiological curvature of the cervical spine. Based on this, a multi-functional cervical vertebra treatment pillow is designed, which not only has the functions of traction, maintaining different positions of the cervical spine, correcting the cervical curvature and equipment exercises, but also has the functions of voice broadcast and network data terminal.

Surgical operation is the main treatment of oral and maxillofacial tumors.Dysphagia is a common postoperative complication.Swal-lowing disorder can not only lead to mis-aspiration,malnutrition,aspiration pneumonia and other serious consequences,but also may cause psychological problems and social communication barriers,affecting the quality of life of the patients.At present,there is no systematic evalua-tion and rehabilitation management plan for the problem of swallowing disorder after oral and maxillofacial tumor surgery in China.Combining the characteristics of postoperative swallowing disorder in patients with oral and maxillofacial tumors,summarizing the clinical experience of ex-perts in the field of tumor and rehabilitation,reviewing and summarizing relevant literature at home and abroad,and through joint discussion and modification,a group of national experts reached this consensus including the core contents of the screening of swallowing disorders,the phased assessment of prognosis and complications,and the implementation plan of comprehensive management such as nutrition management,respiratory management,swallowing function recovery,psychology and nursing during rehabilitation treatment,in order to improve the evalua-tion and rehabilitation of swallowing disorder after oral and maxillofacial tumor surgery in clinic.

Objective : To investigate the effects of actin like 6A (ACTL6A) knockdown on the proliferation, apop⁃ tosis, migration and invasion of SW1990 cells in pancreatic cancer. Methods : The Oncomine database was used to analyze the expression of ACTL6A mRNA in the tissues of pancreatic cancer and normal pancreas. The plasmid of knockdown ACTL6A and siRNA negative control were established and transfected into SW1990 cell line as siRNA⁃ACTL6A group and siRNA⁃NC group. CCK⁃8, cell apoptosis experiment, Wound healing and Transwell assay were used to determine the effects of ACTL6A knockdown on the proliferation, apoptosis, migration and invasion of SW1990 cells. GSEA predicted a possible pathway regulated by ACTL6A in pancreatic cancer. T⁃test was used between the two groups. Results : The expression of ACTL6A in pancreatic cancer tissues was higher than that in normal pancreatic tissues ( P < 0. 05 ) . The results of CCK⁃8 assay showed that the absorbance of siRNA⁃ACTL6A group at 24 and 48 h were lower than those in the siRNA⁃NC group, and the difference was statistically significant ( t = 5. 840, 8. 454, P < 0. 01) . The results of Wound healing assay and Transwell assay showed that the healing rate and the number of invasive cells in siRNA⁃ACTL6A group were both lower than those in the siRNA⁃NC group. The difference was statistically significant ( t = 3. 960,4. 464, P < 0. 05), but the apoptosis rate of siRNA⁃ACTL6A group was significantly higher than that of the siRNA⁃NC group, and the difference was statistically significant( t = 12. 192, P < 0. 001) . GSEA results showed that the group with high expression of ACTL6A mRNA was up⁃regulated in cell cycle, nucleotide excision repair, base excision repair, DNA replication, pathways in cancer, NOTCH signaling pathway and other related gene sets(P < 0. 05) . These pathways were activated when the expression of ACTL6A was up⁃regulated. Conclusion ACTL6A is highly expressed in pancreatic cancer tissues. ACTL6A knockdown promotes the cell apoptosis of SW1990 cells, and inhibits proliferation, invasion and migration of SW1990 cells. The mechanism of the occurrence and development of ACTL6A in pancreatic cancer is attributed to the activation of cell cycle, nucleotide excision repair, base excision repair, DNA replication, pathways in cancer, NOTCH signaling pathway.