Objective To construct recombinant lentiviral vector pSin-EF2-Puro-DAB2IP transfect prostate carcinoma PC3 cells,and to observe its transfection rate and expression level in PC3 cells.Methods The cDNA sequences specifically targeting the DAB2IP gene were designed and cloned into lentiviral vector PC3-pSin using DNA recombinant technique.Using PC3-pSin-EF2 as control,the 293T cells were transfected by Lipofectamine reagent for lentiviral particles packaged, and viral titer was determined. The DAB2IP mRNA and protein expression levels were examined by RT-PCR and Western blotting methods. Results The PCR and sequencing analysis results confirmed that the DAB2IP gene sequence was consistent with the sequence in GeneBank. The number of DAB2IP gene copy in PC3-pSin-EF2-DAB2IP cells and its control PC3-pSin-EF2 cells were 0.001±0.000 and 0.158 ± 0.013,respectively.Compared with control cells,the overexpression of mRNA in PC3-pSin-EF2-DAB2IP cells upregulated (179.370 ± 15.891)times,the difference was statistically significant (P<0.001). Compared with internal reference and control cells, the expression levels of DAB2IP protein in PC3-pSin-EF2-DAB2IP cells upregulated (2.431 ± 0.892)times and (2.415 ± 0.961)times respectively,the differences were statistically significant (P<0.001).Conclusion The lentiviral vector of the DAB2IP gene pSin-EF2-Puro-DAB2IP is successfully constructed,and its targeted gene is stably expressed in the targeted cells,which provides a basis for the further functional study of DAB2IP.

ObjectiveTo establish an experimental model of orthostatic hypotension (OH) after spinal cord injury (SCI) in rabbits and investigate the effect and relevant mechanism of tilting table exercise on OH.MethodsRabbit model of OH was established by completely transecting animal's spinal cord at the T5 segment and putting animal on a 60° tilting table. Tilting table exercise was applied to training group for 28 days starting from 4th day after operation. Systolic blood pressure (SBP) and pulse rate (PR) were observed by non invasive sphygmobolometer of rabbit at 3rd, 10th, 17th, 24th and 31st day after the operation. The concentrations of plasma renin activity (PRA) and angiotensinⅡ (AngⅡ) of different position were detected with radio immunoassay at 31st day after operation.ResultsSBP at 60° head up tilt increased and difference of SBP between two positions decreased significantly in training group at 31st day. The concentrations of PRA and AngⅡ of SCI group and training group were higher than that of sham group. There were no significant differences in the concentrations of PRA and AngⅡ between training group and SCI group.Conclusion T5 complete SCI can induce OH in rabbits. Tilting table exercise can partially improve OH after SCI in rabbit, which may be not related to activation of the circulating renin angiotensin system.

Acupuncture Points ; Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; Female ; Hordeolum ; therapy ; Humans ; Male ; Middle Aged ; Phlebotomy ; methods

Objective To study the correlations between meta moods and clinical behavior of junior college nursing students and provide a scientific basis for improving students' ability of clinical practice.Method The trait meta mood scale(TMMS)and nursing students' clinical practice evaluation scale were used in the study among 120 junior college nursing students.Results The total scores on TMMS and the clinical behavior were(3.45±0.43)and(2.96±0.39),respectively.The scores on the three dimensions of meta moods were positively correlated with 7 dimensions of clinical behaviors and the total score(P<0.01 or P<0.05).Conclusions The meta moods of college nursing students are at high level and their clinical behaviors are at medium level.Teachers and nursing administrators should help students regulate and manage their emotions in order to promote their ability of clinical practice.

Objective To evaluate the efficacy and safety of interventional occlsu ion operait no by analyizn g thes urgical data of 23 csa es of infants with patent ductusa rteriosus complicated with middle and severe pulmo an ry arterial hypertension.Methods Teh 23 cases of infants wiht patne t ductus arteriosus com-plicated with middlea nd severe pulmonary arterial hypertensionw ere collected in the hospital from January 2011 to December2014 .These infatn s rce eived transcateh ter occlusion with intravenuo s anesthesia after the preoperative examination.The operation procse s included:right ventriculography and pulmonary atr ery pressure tested,then lateral angiogar phy of descending aorta was performed to observe the type and size of patent ud ctus atr eriosus and measure ascending aorta,descending aortic pressure,and recorded the pressure re-spectively.1 ml blood sample of ascending aorta,pulmonary artery and inef rior vena vein respectively was used for gas analysis.All these data was used to calculate pulmonary vascular resistance.After tried to plug-ging effectiveyl we can release the occluder.In the postoperative 24 h,1 month,3 months,the infants should be measured with Doppler echocardiography,chest X ray and electrocardiogram examination.Results The clinical symptoms disappeared and the short-term follow-up was not associated with the complications of interventional therapy.Th e comparison of the pressure changes before and after the operation were performed as following, aortic per ssure decreased [ preoperation ( 68.3 ±17.5 )/( 21.4 ±3.7 ) mmHg, postoperation (52.4 ±8.7)/(15.6 ±3.5) mmHg,1 mmHg=0.133 kPa],ascending aorta pressure increased(preoperation (83.5 ±5.9)/(51.3 ±3.6) mmHg,postoperation(88.2 ±5.1)/(52.4 ±2.7) mmHg),and descending aorta pressure increased ( preoperation ( 81.4 ±3.3 )/( 48.2 ±2.7 ) mmHg, postoperation ( 86.5 ±4.7 )/(51.5 ±3.2) mmHg), the differences were statistically significant before and after surgery ( t =5.455/3.945 ,P<0.01;t=-2.696/-1.193 , P<0.05; t=-4.167/-3.745 , P<0.01 ) .Conclusion Under conditions of mastering the appropriate operation time and strengthening the management of the perioperative management,transcatheter measurement is safe and effective for infants with patent ductus arteriosus compli-cated with middle and severe pulmonary arterial hypertension.

OBJECTIVE:To investigate the bacteriological characteristics of nasal secretions of chronic rhinosinusitis(CRS)in our hospital,and conduct drug sensitivity test in order to provide theoretic reference for clinical treatment. METHODS:87 CRS patients underwent nasal endoscopic surgery were selected and the nasal secretions of patients were taken for bacterial culture. Automatic microbial analyzer was used to identify the bacteriological characteristics,and disk diffusion test was adopted for drug sensitivity test. RESULTS:Of 87 patients,bacterial strains were detected from 77 cases of nasal secretions,with positive rate of 88.5%. 112 strains were detected,including 81 strains of aerobic bacterium and 31 strains of anaerobic bacterium,and the most common bacterium were Staphylococcus aureus (20.54%). Antibiotics with the highest sensitivity was moxifloxacin (89.29%), followed by ciprofloxacin(87.50%),vancomycin(87.50%). CONCLUSIONS:The bacterial in nasal secretions of CRS patients has high positive detection rate and mainly are aerobic bacterium. Antibiotics should be used rationally according to the results of drug sensitivity test.

Objective To evaluate the effects of Pidomorph on serum immune cell level and T cell subsets in children with bronchial asthma.Methods 140 pediatric patients with bronchial asthma from January 2014 to October 2016 in our hospital were randomly divided into experimental group and control group according to the order of admission,70 cases in each groups.The control group was treated with conventional glucocorticoid drugs,and the experimental group was treated with pidotimod.The total effective rate of clinical treatment,the rate of rapid breathing in the flow rate,forced expiratory peak value,forced breathing vital capacity were compared between two groups three weeks after treatment,and the serum levels of immune cells(IgA,IgG,IgM)and T cell subsets(CD3+,CD4+,CD8+)were compared before and after treatment.Results The total effective rate was 92.86％(65/70)in the experimental group and 72.86％(51/70)in the control group,the total effective rate in the experimental group was significantly higher than that in the control group(P<0.05); Forced breathing speed in the speed,forced expiratory peak value,forced breathing vital capacity in the experimental group were better than the control group(P<0.05); After treatment,the levels of serum immune cells(IgA,IgG,IgM)in the experimental group were significantly higher than those in the control group(P<0.05); After treatment,the levels of T cell subsets(CD3+,CD4+,CD8+)in the experimental group were significantly higher than those in the control group(P<0.05).Conclusion Pidotimod is effective and safe in the treatment of bronchial asthma in children,it can effectively regulate T cell subsets and improve the immune function of children with asthma.

Objective To investigate the inaccurate causes of platelet(PLT)count and to propose the correcting measures.Meth-ods 180 specimens of abnormal PLT count were collected and divided into 4 groups:low PLT count group (PLT ≤20×109/L,72 cases),microcyte group(54 cases),large platelets group(33 cases)and platelet aggregation group(21 cases).The PLT count was simultaneously performed by using the three ways of the optical method (PLT-O),impedance method (PLT-I)and manual micros-copy method (PLT-M).The detection results were analyzed by the paired t test.Results The PLT count in the low PLT group, microcyte group and large platelets group had no statistical differences between PLT-O and PLT-M(P >0.05),and had statistical differences between the PLT-I and PLT-O(P <0.05).The PLT count of PLT-O in the platelet aggregation group was more close to actual value than that of PLT-I(P <0.05).Conclusion The PLT count by PLT-I has many influencing factors,which are mainly the microcytes interference leading to higher PLT count,missed detection of large PLT leading to lower PLT count,PLT aggrega-tion,etc.When the PLT count appears abnormal,the re-detection should be conducted according to the re-detection regulation, which can be corrected by adopting PLT-M or PLT-O,in necessity,the blood specimen could be re-collected for detection.

Objective To investigate the DNA methylation feature and DNA methylation regulation to its transcription and expression of O6-methylguanine-DNA methyltransferase gene (MGMT) in NaAsO2-treated HaCaT cells. Methods HaCaT cells were treated 72 hours at intervals and repeatedly by 3.13, 6.25,12.50, and 25.00 μmol/L NaAsO2, MGMT gene promoter region was amplified in the transcription initiation site - 329 - + 93 region by bisulfate-sequencing polymerase chain reaction (BSP), the mRNA transcription and the protein expression of MGMT was detected by real-time quantitative PCR and Western blotting. NaAsO2-untreated HaCaT cell was set as a blank control, and human epidermal squamous carcinoma cell strain A431 was set as a positive control. Results Among the groups of HaCaT cells treated with 3.13, 6.25, 12.50 and 25.00 μmol/L NaAsO2, the positive rates of the DNA methylation of promoter region in MGMT gene were 0.63%(l/160), 6.25% (10/160), 10.63%( 17/160) and 18.75% (30/160), respectively, and methylated CpG sites were mainly located in - 249--146 region relative to transcription start site. There was no DNA methylation in the blank control. There were significant differences between the blank control and the NaAsO2-treated cells (x2 = 76.687, P< 0.05). Average levels of MGMT mRNA were 1.518 31 ± 0.180 54, 1.425 22 ± 0.180 39, 1.014 54 ± 0.096 79 and 0.887 72 ± 0.020 00, respectively among the groups of HaCaT cells treated with 3.13, 6.25, 12.50 and 25.00 μmol/L NaAsO2, compared with the blank control cells(1.198 29 ± 0.159 97), there were significant differences(F = 37.359, P < 0.05). Average levels of MGMT protein were 1.174 47 ± 0.064 75, 0.848 83 ± 0.057 01, 0.471 63 ± 0.023 34 and 0.240 34 ± 0.014 43, respectively among the groups of HaCaT cells treated with 3.13, 6.25, 12.50 and 25.00 μmol/L NaAsO2, compared with the blank control cells (1.066 19 ± 0.061 24), there were significant differences(F = 20.687, P < 0.05). Conclusions Arsenic can cause CpC island hypermethylation in the promoter region of MGMT gene, which results in inhibited MGMT mRNA transcription and protein expression. It might be one of the important mechanisms of arsenic-induced skin lesion.

Objective To explore the clinical and electrophysiological characteristics of diabetic lumbosacral radiculoplexus neuropathy (DLRPN). Methods The clinical, electrophysiological and neuroimaging changes in 15 cases of DLRPN were investigated. Results The major clinical manifestations of 13 cases were unilateral or asymmetrical pain and progressive muscular weakness and atrophy of the proximal lower limb. The nerve conduction studies in affected nerves showed absent or reduced compound muscle action potential amplitudes and sensory responses with proportionate slowing of the conduction velocities. F waves and H reflexes were of long latencies or absent. Electromyography of affected muscles showed positive sharp waves and fibrillation potentials involving lumbar paraspinal muscles. Assessment of the motor unit action potentials (MUAPs) revealed high amplitude, long duration, and polyphasic MUAPs with reduced recruitment 50% cases have much lower R-R interval variation. Conclusions DLRPN presents disabling pain and muscular weakness and elctrophysiological examination has a paramount value in the diagnosis and evaluation of this disease