Objective To evaluate the effect of neurotrophin-3 (NT-3) on the expression of serine/threonine protein kinase (Akt) during ropivacaine-induced neurotoxicity to the spinal cord of rats.Methods Healthy adult male Sprague-Dawley rats,aged 1-2 months,weighing 280-320 g,were used in the study.A catheter was inserted at L5,6 interspace into the epidural space of rats.A total of 108 rats,in which intrathecal catheters were successfully implanted,were randomly divided into 3 groups (n =36each):control group (group C),1％ ropivacaine group (group R),and 1％ ropivacaine + NT-3 group (group NT).The equal volume of normal saline was given in group C,1％ ropivacaine 0.12 ml/kg was injected via the intrathecal catheter once every 1.5 h for 8 times in total in R and NT groups.In addition,NT-3 0.1 mg/kg was simultanenously injected via the intrathecal catheter in group NT.On days 1,3,5,7,14 and 28 after the end of administration (T1-6),6 rats were sacrificed in each group.Their lumbar enlargements were removed for determination of neuronal apoptosis (using TUNEL) and Akt expression (by immuno-histochemistry).The apoptotic rate was calculated.Results Compared with group C,the apoptotic rate was significantly increased at T1-4,and Akt expression was significantly up-regulated at T1-3 in group R,and the apoptotic rate were significantly increased,and Akt expression was significantly up-regulated at T1-3 in group NT (P＜0.05).Compared with group R,the apoptotic rate was significantly decreased at T3,4,and Akt expression was significantly down-regulated at T2.3 in group NT (P＜0.05).Conclusion The mechanism by which NT-3 reduces ropivacaine-induced neurotoxicity to the spinal cord may be related to down-regulation of the expression of Akt in rats.

In view of the problems existing in medical image network teaching, such as redundant and rigid curriculum system, obsolete structure, untimely updates, lack of supervision system and poor stability of network security and so on, we explored how to improve the medical image network teaching from the fusion of traditional teaching, optimizing curriculum system, perfecting the medical imaging system of autonomous learning, encouraging online communication, creating a good atmosphere, increasing invest-ment in software development and multi-sectoral collaboration, in order to complete the transfer of knowl-edge more effectively, and promote the good development of medical imaging teaching.

Objective To screen the conservative,stable and specific DNA signature sequence in the plasmid of Yersinia pestis.Methods Specific validation trials and stability of the qualification test were carried out to 40 strains of Yersinia pestis,47 strains of non-Yersinia pestis of home and wild types of rodent in Yunnan,by using 32 DNA sequences derived from Yersinia pestis in the plasmid and conventional PCR technology,and Yersinia pestis vaccine strain EV76 as a positive control.Results Four pairs of relatively conservative,stable and specific DNA marker genes were screened:YPMT1.05c,YPMT1.03c,YPMT1.42 and YPMT1.04c.Conclusions The 4 pairs of Yersinia pestis DNA signature sequences can be used for rapid diagnosis of plague.

Objective To investigate DNA double-strand breaks and radiosensitization in renal carcinoma 786-O cells induced by fludarabine (FA) combined with different ionizing radiations.Methods The 786-O cells were exposed to FA combined with X-ray or heavy ion beam irradiation.Flow cytometry was used to evaluate the percentage of γH2AX-positive cells and cell cycle.The neutral comet assay was used to detect DNA double-strand breaks.The colony-forming assay was used to evaluate the effects of different treatments on cell survival.Comparison between groups was made by one-way analysis of variance or Dunnet' s t test.Results Compared with FA alone or irradiation alone,FA combined with different ionizing radiations increased DNA double-strand breaks as shown by significantly increased levels of γH2AX (P=0.007,0.001);FA combined with heavy ion beam irradiation lead to a cell cycle block at the radiosensitive G2/M phase and significantly increased the expression of γH2AX in the G2/M phase (P=0.000,0.000);the neutral comet assay revealed that FA combined with irradiation significantly increased DNA sublethal damage (P=0.020,0.060);FA significantly reduced the colony-forming rate after irradiation (P=0.000,0.030;0.001,0.040).Conclusions FA enhances the effects induced by X-ray and heavy ion beam irradiation with different properties.Particularly,FA substantially enhances the cell death induced by heavy ion beam irradiation.

Targeted therapeutic strategy for cancer stem cells (CSCs) is the key to prevent tumor relapse and metastasis. The im-portant roles of epigenetic regulation on the development of stem cells and gene reprogram of somatic cells suggest that this process may remarkably affect the occurrence and development of CSCs. The epigenome, which comprises DNA methylation, histone modifica-tions, chromatin structures, and non-coding RNAs, controls gene expression patterns. In renal cell carcinoma (RCC), aberrant changes occur in the epigenome. To date, cells with CSC properties from RCC have been successfully isolated using different methods, such as sorting using the Hoechst 33342 side population, forming tumor spheroid, and sorting CD105 cell surface biomarker. According to the progress in genetic studies on RCC, in addition to DNA sequence, the abnormality in the regulatory mechanism has considerable func-tions in tumor progression. Epigenetic changes may be integral to the behavior of cancer progenitor cells and their progeny. Knowledge on epigenetics in renal tumorigenesis process is beneficial in the development of new therapeutic modalities and may deliver new prog-nostic and early diagnostic markers. This paper reviews the latest development in the study of RCC stem cells and the underlying mech-anisms of epigenetic regulation on the development of CSCs in RCC.

Objective To investigate the correlation between leukoencephalopathy and atherosclerosis. Methods 238 patients were enrolled. All the patients were underwent brain MRI and carotid and femoral artery duplex ultrasonography after hospitalized. The atherosclerotic plaque, intima-media thickness (IMT), peak systolic velocity (Vp) were measured with ultrasonography. The data was analyzed with SPSS 13.0 software. Results The leukoencephalopathy of the patients positively correlated with age, hypertension, serious of atherosclerosis and carotid IMT, negatively correlated with carotid Vp, but did not correlated with vertebrarterial Vp. Conclusion The leukoencephalopathy positively correlate with some atherosclerotic risk factors, the serious of atherosclerosis, and anterior circulation disorder, but do not correlate with posterior circulation.

The sfa1 gene encoded a bifunctional enzyme with the activities of both alcohol dehydrogenase and glutathione-dependent formaldehyde dehydrogenase in Saccharomyces cerevisiae.The gene disruption cassette produced by PCR using the same long oligonucleotides which comprise 19 or 22 nucleotides complementary to sequences in the templates(pUG6 and pUG66 marker plasmid)at 3' end and 45 nucleotides at 5' end that annealed to sites upstream or downstream of the genomic target sequence to be deleted.After two linear disruption cassettes with a Cre/loxP mediated marker were transformed into the cells of Saccharomyces cerevisiae YS-1,the positive transformants were checked by PCR to correct the integration of the cassette and concurrent deletion of the chromosomal target sequence.Once correctly integrated into the genome,the select marker can be efficiently rescued by transformating the plasmid pSH47 into YS-1 and inducing the Cre expression with a Cre/loxP-mediated marker removal procedure.The expression of the Cre recombinase finally resulted in the removal of the marker gene,leaving behind a single loxP site at the chromosomal locus.The diploid mutant YS-1-sfa1 was generated,which could enhance the output of ethanol with 8.0% by shaking culture in flask compared with the original strain YS-1.

Objective To investigate the effects of gastric bypass on glycometabolism and improvement of islet β cell function and insulin resistance in patients with type 2 diabetes. Methods Eight patients with type 2 diabetes combined with gastric carcinoma who treated with gastric bypass were studied prospectively. Fasting and postprandial plasma glucose levels, fasting and postprandial insulin C-peptide levels, and body mass index (BMI) were measured right before the surgery and at intervals of 1 week, 2 weeks, 1 month and 3 months after the surgery. Glycosylated hemoglobin (HbA1c) levels were measured before and 3 months after the surgery. The outcome of the diabetes after 3 months of the surgery was also monitored. Results Fasting and postprandial plasma glucose levels decreased (P ＜ 0.05) and fasting and postprandial insulin C-peptide levels increased (P ＜ 0.05) after the surgery. HbA1c levels also decreased (P ＜ 0.05) after 3 months of the surgery. There was no significant change of BMI at all intervals after the surgery(P＞ 0.05). All of the 8 patients reached the total effective standard and 6 patients reached the clinical remission standard after 3 months of the surgery. Conclusions It suggests that gastric bypass can significantly lower plasma glucose levels in type 2 diabetes, which does not depend on the loss of weight. The control of plasma glucose by gastric bypass may be due to the improvement of islet β cell function and increasing secretion of endogenous insulin.

Objective: To observe the clinical effects of Shu-acupuncture method in Nei Jing (Classic of Internal Medicine) in the treatment of shoulder and arm pain. Methods: A total of 90 patients with shoulder and arm pain were randomly divided into an observation group and a control group, 45 cases in each group. The treatment of Shu-acupuncture method in Nei Jing (Classic of Internal Medicine) was adopted in the observation group, routine acupuncture was used in the control group. The two groups were treated once every day, with 5 treatments as one course, and a 2-day rest between two courses. After 3 courses, pain was assessed by visual analog scale (VAS), and the clinical effects were compared between the two groups. Results: After the treatment, VAS scores were significantly changed in both groups (both P<0.01). The VAS score was lower in the observation group than that in the control group, with a statistical difference between the two groups (P<0.05). The total effective rate was 100% in the observation group, versus 91.1% in the control group, the difference was statistically significant (P<0.05). Conclusion: The therapeutic effect of Shu-acupuncture method in Nei Jing (Classic of Internal Medicine) is better than that of routine acupuncture in treating shoulder and arm pain.

Objective To construct a lentiviral vector of RNA interference (RNAi) of PKCγ gene. Methods The effective sequence of siRNA targeting PKCγ gene was confirmed in our previous study. The complementary DNA containing both sense and antisense oligo DNA of the targeting sequence was designed, synthesized and cloned into the pGCSIL-GFP vector, which contained U_6 promoter and green fluorescent protein (GFP) . The resulting lentiviral vector containing PKCγshRNA was named lentivinis RNAi vector of PKCγ, and it was confirmed by realtime PCR and sequencing. 293T cells were cotransfected with lentiviral vector pGCSIL-CTP, pHelper 1.0 and pHelper 2.0. All virus stocks were produced by calcium phosphate-mediated transfection. The titer of virus was tested according to the expression level of GFP. Results PCR and DNA sequencing demonstrated that the lentivirus RNAi vector of PKCγ producing PKCγshRNA was constructed successfully. The titer of concentrated virus was 1 ×10~9 TU/ml. Conclusion The lentivinis RNAi vector of PKCy was constructed successfully.