RNA interference(RNAi)is a post-transcriptional gene-silencing phenomenon which is triggered by double-stranded RNA(dsRNA).Attempts age being made to exploit RNAi in a clinical setting.However,before RNAi can be exploited as therepeutically,several obstacles must be overcome such as the efficient and safe delivery of siRNA into target tissues and cells.Any usefull delivery method should be desinged to target siRNA.

Objective To construct and express ribonuclease-resistant virus-like particles containing long chimeric RNA sequence by changing quantity and affinity of ms2 19mer stem-loop.Methods 1.7 kb maturase and coat protein gene of ms2 were amplified.We used a C-variant aptamer of the wild-type stem-loop where uridine-5 has been substituted by cytosine.The 1700 bp PCR-amplified DNA fragments was gel-purified and then digested with BamH Ⅰ and Hind Ⅲ and ligated to linearized pET28b vector to generate recombinant plasmid pET-MC-pac.The five target DNA sequences were spliced by overlapping extension(including 3 fragment of SARS-CoV,one fragment of HCV and one fragment of HSN1).PCR was carried out using primers contained Not Ⅰ site and then PCR were cloned into a pGEM(R) T-easy vector and then excised with the Not Ⅰ restriction enzymes from the resulting recombinant plasmid.simoutaniously the pET-MC-pac plasmid was digested with Not Ⅰ restriction enzymes,fragments were ligated into the linearized pET-MC-pac plasmid to produce a new donor plasmids pET-MC-3V.Simoutaniously,we constructed three recombinant expression vector for control,including N-P3V-pET-P、N-P3V-pET-Cand P-3V-pET-P.N-P3V-pET-P contained one wild type pacsite located behind ms2 sequence:N-P3V-pET-C contained one C-variant pacsite located behind ms2 sequence:P-3V-pET-P contained two wild type pacsites that one is located behind ms2 sequence,another is located between SARS-CoV 3 and HCV sequence.Then these four expression vector were transformed into E.coli strain BL21(DE3),respectively.The 3V Armored RNA was expressed and purified.A260 absorbance value of expression product was determined.Results Four expression plasmids were constructed successfully.The Armored RNA with 1891 bases was snccessfully expressed by the two-paesite expression system of pET-ms2-3V and P-3V-pET-P;for N-P3V-pET-P、N-P3V-pET-C,only 1 200 bp target sequence was packaged into virus-like particles.Expression efficiency of N-P3V-pET-P、N-P3V-pET-C、P-3V-pET-P and N-P3V-pET-P was 0.23,0.35,0.35,0.51 mg/ml,respectively.Armored RNA was shown to have the charaeterization of Ribonuclease-resistant and stable at 4℃,37℃ and 25℃ respectively.Conclusion The expression plasmids containing two pacsites were constructed successfully and prokaryotic expression system can be used as an expressing plasmid platform for preparation of Ribonuclease-resistant virus-like particles containing long chimeric RNA sequence.

Objective To investigate current situation of teaching competition and to put forward some feasible suggestions for improving the teaching competition of theoretical course for young teachers. Methods Questionnaire survey was conducted for 126 teachers including the effect of teaching competi-tion on effective teaching atmosphere,comprehensive teaching results,teaching ability and recommenda-tions for improvement. SPSS software was used to do statistical analysis andχ22 test was applied. P<0 . 05 stands for statistically significant difference. Results Investigation results demonstrated that teaching competition of theoretical course was conducive to forming effective teaching atmosphere,achieving better teaching effect and improving teaching ability of teachers . A χ22 test was performed on the cognition of the cultivating of classroom management ability;results demonstrated that there were significant differ-ences between teachers with more than 10 years education age and those with less than 10 years education age(χ22=4.375,P=0.0365). Conclusions Teaching competition of theoretical course is an effective way to cultivate young teachers teaching ability. Teaching competition should be combined with training and feedback and evaluation should be more scientific and objective in order to improve young teachers' teaching ability.

Objective To compare the procotol of EGDT +Pt (cv-a ) CO2 with EGDT in fluid resuscitation and management after evaluate fluid responsiveness in shock patients by Vigileo and passive leg raising test.Metheds Prospectively collect patients who meet the criteria between 2013.5.1-2013.1 1.30 in our ICU.Randomly (random number)divided into Vigileo group (first evaluate the fluid responsiveness then give EGDT +Pt (cv-a) CO2 ) and CVP group (give EGDT).Compare the hospital mortality and morbility of MODS,the volume given in the first 6 hours and the first 7 days,consume of blood products , the ICU and hospital stay.Results Collected 46 patients,Vigileo group (21 )and CVP group (25 ). There’s no significant difference between groups at baseline.In the first 6 hours the CVP group had received more fluids (3656.281678.57 vs. 2639.141326.59 ) mL, P =0.03;and more blood products (573.00172.57 vs. 190.4770.82)mL,P=0.04,respictivily.Vigileo group significantly short the ICU stay ,(6.384.34 vs. 12.165.77)d,P=0.04.But there’s no significant difference in hospital motality and the morbility of MODS.The ROC of Age ,the accumulative volume of balance in 7 days,APACHEⅡscore in the first day to predict death is 0.84 (0.68-0.99)、0.82 (0.69-0.95)、0.80 (0.66-0.94),all P>0.05,respectively.By 7 days the accumulative volume of balance 3454.51mL as cutoff to predict death with the sensitivity of 0.67,specificity of 0.84.Conclusions 1.Given EGDT +Pt (cv-a) CO2 after evaluate the fluid responsiveness can reduce fluid and blood products given in the first 6 hours,significantly short the ICU stay,without worsen the tissue flow or increase the morbility of MODS;2.Consecutive positive fluid balance is a risk factor about poor prognosis,and also a sensitive indicator to predict death.

Objective To observe if fluoxetine has a potency to inhibit the progression of Lewis lung cancer by combining the fluoxetine and cisplatin to treat the mice bearing Lewis lung cancer with depression .Methods We devel-oped a mouse model of Lewis lung cancer with depression which was intervened with cisplatin and fluoxetine , and the indi-cators related to cancer and depression were tested .Social interaction test was used to measure the behavioral changes of the depressed model mice .The serum cortisol and IL-6, BDNF mRNA in the hippocampus , and NF-κB and VEGF in the tumor tissue were selected for investigation and comparison .Results The mice which were induced by social defeat exhib-ited social avoidance behavior in the social interaction test .The cortisol and IL-6 level in both combination groups was de-creased compared with that in the model group (P<0.05), and the cortisol and BDNF mRNA in the combination group in-creased signifuicantly ( P <0.05 ) .There were no statistically significant differences in the tumor weight , NF-κB and VEGF in tumor tissues between the single and combination groups (P>0.05).Conclusions Fluoxetine has antidepres-sant effect by decreasing the high level of serum cortisol and promoting the BDNF mRNA expression in the hippocampus . However , fluoxetine is not found to have the potency to inhibit the expression of NF -κB related with the progression of tumor.

Objective To evaluate the effectiveness of rehabilitative treatment for severe cervical spondylotic myeiopathy after combined posterior-anterior surgery. Methods Fifty-four patients (mean age, 59.5) who had undergone combined posterlor-anterior surgery of the cervical spine due to severe cervical spondylotic myelopathy were studied with an average follow-up of 38.7 months. All the patients were allocated into either a rehabilitation treatment group (28 patients) or a control group (26 patients). Neurological function in the two groups was assessed using the Japanese Orthopedic Association (JOA) grading system. In addition, the status of axial symptoms, wound healing and post-operative complications were also evaluated. Results The bone grafts completely fused in both groups. All of the wounds in the rehabilitation treatment group healed without any infection. Three wounds were infected in the control group, but were cured by prompt treatment. Two patients in the rehabilitation group and 9 in the control group suffered laryngeal edema. There was no significant difference between the two groups before the operation with regard to their JOA scores, which significantly increased postoperation in both groups. In the rehabilitation treatment group, the postoperation JOA scores were significantly higher than in the control group. At the same time, there were significantly fewer patients with neck axial symptoms in the rehabilitation treatment group than that in the control group (25.0% versus 69.2% , P < 0.01 ). Conclusions Systematic rehabilitation treatment pro-and post-operation of patients with severe cervical spondylotie myelopathy can accelerate neurological recovery and help prevent postoperative complications and neck axial symptoms.

Twelve ASA Ⅳ or Ⅴ patients with severe airway stenosis, aged 34-87 yr, weighing 50-80 kg,were enrolled in the study. The guidewire and endotracheal tube were inserted through the oral cavity under the guidance of the method of interventional radiology. The expandor was then exchanged and the endotracheal tube was placed in the suitable position within the trachea along the expandor under X-ray guidance. Endotracheal intubation was technically successful in all patients, without serious complications in the process. The intubation time was 1-5 min.

Objective To investigate the appropriate surgical approach in the management of cervical cord injury following ossification of the posterior longitudinal ligament. Methods The clinical data of 25 patients with cervical cord injury following ossification of the posterior longitudinal ligament who received surgical treatment were retrospectively analyzed. According to Frankel grades, two patients were at grade A, three at grade B, 14 at grade C and six at grade D. The surgical procedures consisted of anterior decompression (12 patients), posterior decompression (8 patients) and combined posteroanterior decompression (5 patients). Results No iatrogenic injury of great vessels, trachea, esophagus or spinal cord occurred. All the patients were followed up for 15-86 months (mean 38.3 months). All segments with anterior fixation attained solid fusion, without implants loosening or breakage. No reelosed open-door was found in patients who received posterior laminoplasty. The spinal function got improved in 21 patients, and a relief of pain or numb of the upper limb was attained in four patients whose spinal cord injury was not cured. Conclusions The surgical outcome of cervical cord injury following ossification of the posterior longitudinal ligament is satisfactory. It is important to select a suitable surgical approach according to the imaging manifestations associated with the general conditions of the patients.

OBJECTIVETo evaluate the methods used to diagnose and surgically treat ruptured aneurysm in sinus of Valsalva (RASV).

METHODSThirty-seven hospitalized patients with ruptured aneurysms in the sinus of Valsalva from September 1981 to April 2001, including 21 cases (56.7%) of RASV associated with ventricular septal defects (VSD) and 11 (29.7%) with aortic valvular prolapse were given surgical interventions. Under hypothermia and extracorporeal circulation, we successfully performed the surgical correction of RASV for all 37 patients VSD repair in 21 patients, aortic valvuloplasty in 6 and aortic valvular replacement in 2.

RESULTSThere was no hospital deaths among these patients, although residual shunting occurred in two patients and acute renal failure was found one. Follow-up study of one month to 20 years in the patients undergoing repair of RASV revealed that the mostly individuals treated with operation obtained satisfactory cardiac function.

CONCLUSIONCorrect diagnosis of ruptured aneurysm in sinus of Valsalva should be confirmed immediately and surgical correction should be carried out as soon as possible.

Adolescent ; Adult ; Aortic Aneurysm ; diagnosis ; surgery ; Aortic Rupture ; diagnosis ; surgery ; Child ; Female ; Humans ; Male ; Middle Aged ; Sinus of Valsalva

objective To understand the baseline data of mortality from malignant tumor from 2006 to 2009 around Hongyanhe Nuclear Power Plant in Wafangdian City,Liaoning Province,so as to provide scientific basis for evaluating the impact of normal operation of nuclear power plant on the health of the residents nearby.Methods Thirty small towns near Hongyanhe Nuclear Power Plant were divided into 5 investigated areas according to the distances away therefrom(0.,10.,20-,30-,and 40-km).The data about from malignant tumor were obtained from the Center for Disease Control and Prevention of Wafangdian.The mortality distribution of difierent malignant tumors was analyzed,including the radiosensitive malignant tumors,especially leukemia,breast cancer,and thyroid gland cancer in different area,gender,and age groups.Results The mortality from malignant tumor was 151.97/105,and the standardized mortality rate(SMR)was 97.76/105.The mortality from malignant tumor among the males was 188.28/105(with the SMR of 116.76/105),and that among the females was 113.47/105(with the SMR of 75.89/105).with a sex ratio of 1.71.The first five cancers in the rank of death causes were lung,liver,stomach,colorectal,and esophageal cancers with mortality of 46.19/105,23.51/105,20.30/105,8.06/105 and 5.45/105,respectively.The mortality from mal.ignant tumor in the areas around the nuclear power plant from the near to the distant were 99.85/105, 137.40/105,138.73/105,156.30/105,and 154.16/105,respectively.The mortality from radiosensitive malignant tumors,leukemia,breast cancer,and thyroid gland cancer were 4.57/105,4.06/105,and 0.26/105,respectively.Conclusions Lung cancer and digestive tract malignant tumors are the main causes of death from malignant tumors in Wafangdian area before the nuclear power plant began to operate.There are no significant differences in the mortality distribution of malignant tumors among different areas,genders,and age groups.There are not significant differences in the mortality distribution of leukemia and breast cancer among different areas and age groups.