Objective.To probe into the influence of Jiannao Anshen Capsule of Miao medicine on the behavioral performance,content of cytokine interleukin-1 (IL-1),neuroendocrine hormone melatonin (MT) and monoamine neurotransmitter 5-hydroxytryptamine (5-HT) of rats with sleep deprival.Methods All of the 60 rats were evenly and randomly divided into 5 groups:the normal control group,model control group,estazolam group,Zaoren Anshen group,and Jiannao Anshen of Miao medicine group,respectively.Every group had 12 rats.Sleep deprivation rat model was established by intraperitoneal injection of para-chlorophenyla lanice (PCPA).After successful modeling,the rats were intragastrically administrated with corresponding medicines.The contents of serum 5-HT,IL-1 and MT and the contents of 5-HT,MT in brain tissue were determined.Results Behavioral score,the contents of 5-HT,MT and IL-1 in serum and the contents of 5-HT,MT in brain tissue in insomnia rats caused by PCPA were significantly decreased as compared with normal control group (P ＜ 0.05).In the Jiannao Anshen group,estazolam group and Zaoren Anshen group,the behavioral score,the content of 5-HT,MT and IL-1 in serum and the content of 5-HT and MT in brain tissue of insomnia rats were significantly increased as compared with model control group (P ＜ 0.05).The behavioral score,the content of 5-HT,MT and IL-1 in serum and the content of 5-HT and MT in brain tissue of insomnia rats were not statistically significant different among Jiannao Anshen group,estazolam group and Zaoren Anshen group (P ＞ 0.05).Conclusion Jiannao Anshen capsule of Miao medicine could obviously improve the sleep quality of rats with sleep deprival.The mechanism may be related to improving the content of IL-1,MT and 5-HT.

Objective To compare the clinical effect of radionuclide myocardial perfusion imaging(SPECT)and multilayer spiral CT coronary artery imaging(MSCT) in the diagnosis of coronsry artery disease(CHD).Methods The clinical data of 36 patients suspected with CHD and routine inspection of SPECT and MSCT were retrospectively analyzed.The diagnostic effects of two methods were compared.Results The sensitivity,specificity and the rate of accuracy of SPECT was 89.47％,94.12％,and 91.67％,respectively,while the sensitivity,specificity and the rate of accuracy of MSCT was 84.21％,94.12％,and 88.89％,respectively.There were no obvious difference in two groups (x2 =1.265,1.668,2.005,1.526,1.889,all P ＞ 0.05 ).Conclusion There was a high degree consistency on the diagnosis of CHD between SPECT and MSCT.In the application,combined with the two methods could improve the diagnosis of CHD.

Objective To explore the therapy efficacy for children with primary nocturnal enuresis by meta-analysis on the efficiency of alarm treatment versus desmopressin.Methods PubMed, Central, Elesvier, CNKI and some other databases were browsed to obtain all randomized controlled trails(RCT) and to compare the therapy between alarm treatment and desmopressin.Data extraction and quality evaluation were done by methods recommended by Cochrane center.The results of short-term and long-term efficacy and compliance were analyzed by Review Manager 5.0.Results Fifty-four RCT were retrieved and 11 RCT were included in the study.There was no statistical difference between alarm treatment and desmopressin when analyzing by no wetting episode,wetting no more than 1 night per month,wetting nights' decrease over 90％ or wetting nights decrease over 50％.By 3-month follow-up after treatment stopped,alarm treatment significantly reduced the wetting nights to over 50％ or less than 1 night per month than desmopressin.By 6-month follow-ups after treatment stopped, there was no difference between alarm treatment and desmopressin when analyzed by wetting no more than 1 night.The relapse rate of desmopressin was higher than that of alarm treatment (P =0.007).However, the withdrawal and abandonment rate of alarm treatment was higher than that of desmopressin(P ＜0.000 01).Severe adverse effects were not found in both of the treatment groups.Conclusions There is no statistical difference between alarm and desmopressin therapy in short-term treatment.The efficiency of alarm device is better than that of desmopressin in long-term treatment.The follow-up of alarm treatment lost more than that of desmopressin.Desmopressin is better than the alarm treatment in compliance but has higher recurrence rate.

Background Meningothelial cells (MECs) occupy the predominant cell component of barrier between optic nerve and the cerebral spinal fluid,and any change of cerebral fluid components probably affects the MECs function and further impairs the optic nerve.Objective This study was designed to investigate the influence of glutamate,a potentially excitotoxic amino acid,to the functional changes of MECs and provide a theoretical evidence for clarifying the mechanism of optic nerve disorders.Methods Human MECs strains were cultured in vitro and prepared into cell suspension.The cells were inoculated to 96-well plates with the densities of 1 × 104/we11.The glutamate of 100,200,400,600,800 and 1 000 μmol/L was added into medium for 12,24,36,48 and 72 hours,respectively,and the cultured cells without glutamate were used as normal control group.MTS assay was employed to measure the proliferative rate (absorbency) of the cells.The regularly cultured MECs were divided into 600 μmol/L glutamate-treated group and normal control group and the cells were treated for 12 and 24 hours respectively,and the expression of superoxide dismutase (SOD) mRNA and heat shock protein 90 (HSP90) mRNA in the cells was detected by real-time PCR;the level of total anti-oxidative capacity (T-AOC) of the cells was processed by enzyme linked immunosorbent assay (ELISA),and the reactive oxygen species (ROS) production was determined by DCFH-DA probe.Results Cultured MECs grew well and formed 80％ confluence after 72 hours culture.The proliferative rate of the cells were gradually decreased with the increase of glutamate dose and the lapse of affected time,with significant differences among different concentrations of glutamate and various time points (F tration =52.501,P＜0.001;Ftime =8.505,P＜0.001).The relative expression level of SOD mRNA was significantly reduced in the glutamate-treated group compared with the normal control group in both 24 hours and 48 hours after culture (t =20.278,t =16.724,both at P＜0.001),and the expression of HSP90 mRNA in the cells was significantly lower in the glutamate-treated group than that in the normal control group in 24 hours after culture (t =5.065,P =0.002).No significant difference was found in T-AOC activity between glutamate-treated group and normal control group in 24 hours after culture ([30.835±2.094] nmol/(min · L) vs.[32.873±2.317] nmol/(min · L)) (t=1.599,P =1.414).In 48 hours after culture,T-AOC activity was (29.561 ± 1.831) nmol/(min · L) in the glutamate-treated group,which was significantly lower in comparison with normal control group (33.680±2.039) nmol/(min · L)(t =3.682,P =0.004).Fluorescence staining showed that the intensity of green fluorescence of ROS in MECs in the normal control group was weaker than that in the glutamate-treated group under the immunofluorescense microscope.The ROS level was 48.110± 1.712 and 40.982± 1.853 at 24 hours and 48 hours in the glutamate-treated cells,and which was significantly elevated in comparison with 36.608± 1.009 and 37.153 ± 1.424 in the normal control group (t=14.178,P＜0.001;t=4.012,P=0.002).Conclusions Glutamate inhibits the proliferation of MECs in vitro,and excitatory toxicity of glutamate on MECs probably is associated with oxidative stress response.

To observe the alteration in the expression of DNA repair enzymes hOGG1 and hMYHalpha and the change in 8-OHdG levels in the HBx gene-transfected cells HepG2/HBx and to explore the mechanisms of the HBV-associated hepatocellular carcinoma, the gene-transfected cells HepG2/HBx which stably expressed HBx was established, and the effect of HBx on the cell cycle and proliferation of HepG2 was examined. By using the beta-actin as the interior control, real-time polymerase chain reaction (Real-time qPCR) was employed to quantitatively detect the expression of DNA repair enzymes hOGG1 and hMYHalpha in the HepG2/HBx, the control cells HepG2 and HepG2 transfected with pcDNA3.1 vector (HepG2/pDNA3.1). The 8-OHdG levels were determined by HPLC/ECD in the established gene-transfected cells HepG2/HBx and the control cells HepG2 and HepG2/pcDNA3.1. Our results showed that the expression of DNA repair enzyme hMYHalpha in the HepG2/HBx (0.021+/-0.007) was significantly lower than that of HepG2 (0.099+/-0.041) (P<0.05) and HepG2/pDNA3.1 (0.121+/-0.005) (P<0.05). However, the no significant differences existed in the expression of DNA repair enzyme hOGG1 among the three cell strains (P>0.05). The 8-OHdG level in the HepG2/HBx was significantly higher than that in HepG2 and HepG2/pcDNA3.1 (P<0.05). It is concluded that HBx gene may inhibit the expression of DNA repair enzyme hMYHalpha mRNA to impair the ability to repair the intracellular DNA oxidative damage, to increase the oxidative DNA-adduct 8-OHdG and to affect the nucleotide excision repair function, thus participate in the occurrence and development of hepatocellular carcinoma.
DNA Glycosylases/genetics ; DNA Glycosylases/*metabolism ; Deoxyguanosine/analogs & derivatives ; Deoxyguanosine/metabolism ; Hep G2 Cells ; RNA, Messenger/genetics ; RNA, Messenger/metabolism ; Trans-Activators/*genetics

Objective:To investigate the relationship between primary resection and KRAS gene mutation in the mild symptomatic patients with stage Ⅳ colorectal cancer,and to clarify its significance of prognosis.Methods:The clinical data of 46 mild symptomatic patients with stage Ⅳ colorectal cancer 2010 to December 2010 were collected.All the patients received primary resection.The KRAS gene mutation in the patients was detected by direct sequencing and the patients were followed up for 5 years.The influence of primary resection and KRAS gene mutation in prognosis of the patients with stage Ⅳ colorectal cancer was analyzed, and the clinical pathological features which might influence the prognosis were analyzed by survival analysis.Results:In 46 patients with colorectal cancer, KRAS gene mutation was found in 20 cases, the mutation rate was 43.4%, and most mutation was found at Codon 12. The KRAS mutation had relationship with the tumor site and multiple metastasis (P<0.05).The Kaplan-Meier survival curve and univariate analysis results showed that the median survival time of the patients with wild type of KRAS gene was 58.4 months, the median survival time of the patients with mutant type of KRAS gene was 42.2 months, and there was no significant difference (P>0.05).The median survival time of right colon cancer patients was 34.2 months, the median survival time of left colon cancer patients was 58.3 months, and there was sigificant difference (P<0.05).The cancer metastases including liver, lung and multiple metastasis were closely related to the poor prognosis of the colorectal cancer patients(P<0.05).The median survival time of the patients with stage Ⅳ colorectal cancer was 39.6 months after operation.Conclusion:After primary resection of the mild symptomatic patients with stage Ⅳ colorectal cancer,the median survival time of the patients with colorectal cancer in left colon site and right colon site were prolonged.Right colon cancer has more poorer prognosis than left colon cancer.KRAS gene mutation is associated with the tumor site and the multiple metastasis.The location of metastasis affect the prognosis.

The incorporation of site-specific recombination systems can help to overcome bottlenecks in livestock transgenic technology. For evaluating the efficiency of Cre/lox mediated DNA recombination in embryos and somatic cells, a working model was established using rat mammary carcinoma cell line SHZ-88, aimed at creation of and use repeatedly of selected "friendly loci" in transgenic livestock. An integration vector pTE-lox2272-DsRed-loxP-GFP-loxP, which red fluorescence gene DsRed served as the first target gene and green fluorescence gene GFP as marker gene, was constructed for introduction of acceptor loci in genome. At the same time a replacement vector pT-lox2272-neo-loxP in which Neo coding sequence served as the second target gene was also constructed for replacing DsRed gene. Transgenic cell clones were produced by electroporating SHZ-88 cell with the integration vector. Cells from three transgenic clones selected randomly were further amplified and were then co-electroporated with the replacement vector as well as cre gene. Analysis of the expression patterns of DsRed and GFP indicated that among the 1 070 cell colonies the efficiency on marker GFP deletion was 91.1% and the efficiency on gene replacement was 29.3%. Molecular analysis by PCR and Southern blotting confirmed that the color patterns as expressed by cell colonies could represent the actual molecular events. This working model mediated by Cre/lox system should be useful for the improvement of the present animal transgenic technology.

Objective To analyze the efficacy,late complications and prognostic factors of post-op-erative radiotherapy for cervical cancer. Methods From Nov. 1999 to Feb. 2005,114 patients with cervi-cal cancer received adjuvant pelvic radiotherapy after radical hysterectomy and pelvic lymphadenectomy. The median age was 42.5 (24 to 72) years old. According to the FIGO staging system,6,51,18,26 and 13 pa-tients had stage ⅠA, Ⅰb1, Ⅰb2, ⅡA and ⅡBdisease. The pathological type was squamous cell carcinoma,ade-nocarcinoma, squamous-adenocarcinoma and undifferentiated carcinoma in 92,19,2 and 1 patients, respec-tively. The whole-pelvic external beam irradiation of 50 Gy (40 to 60 Gy) was given with 6 MV or 15 MV X-ray beams using four-field box technique. Eighty-one patients received intravaginal brachytherapy of 16 Gy (4 -30 Gy in 1 -6 fractions) 4 weeks after the beginning of radiotherapy, with the referrence point being at 0.5 cm under the vaginal mucosa. Eighty-seven patients received preoperative and/or concurrent chemother-apy. The survival and independent prognostic factors were analyzed by Log-rank method and Cox model. Results The median follow-up time was 26.0 (5 - 75) months. The overall survival rate, disease-free sur-rival rate and local control rate were 93.1%, 88.1% and 94.6% at 2-year, and 75.7%, 62.3% and 85.6% at 5-year,respectively. The independent prognostic factors were lymph node metastasis and positive surgical margin for overall survival, positive surgical margin for local control, and stage, uterine body invasion and positive surgical margin for disease-free survival. Sixteen patients ( 14% ) had distant metastasis, and the most common sites were the lung,inguinal region,bone,liver and brain. According to RTOG grading sys-tem, the incidence of late gastrointestinal side effects of grade 1,2 and 3 was 11.4%, 11.4% and 3.5%. The corresponding genitourinary side effects were 14.0% ,6.1% and 0.9%, respectively. The incidence of leg lymphedema was 7 % . Conclusions Post - operative radiotherapy can achieve good local control in cervical cancer with acceptable late side effects. Distant metastasis is the main cause of death.

Objective To investigate the perioperative nursing experience for patients with mayer rokitansky kuster hauser (MRKH).Methods We conducted multidisciplinary care to a MRKH patient,including personalize psychological support,bowel preparation,two surgical nursing convergence and vaginal mold care during hospitalization.Results The patient discharged after clinical symptoms remission,the patient sexual life was satisfaction,no complications were found in the six months follow-up.Conclusion The key of operation success is good coordination and cooperation of multiple departments,close observation and effective nursing in the perioperation period.