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 investigate the clinical efficacy of lengthened proximal femoral nail antirotation (PFNA) and Gamma nail treatment of proximal femoral fractures (including the intertrochanteric and inferior intertrochanteric area) combined with ipsilateral femoral shaft fractures.Methods A retrospective case control study was made on 68 cases of proximal femoral fractures combined with ipsilateral femoral shaft fractures received from January 2013 to October 2014.According to the operation mode,the patients were assigned to PFNA group (30 cases) and Gamma nail group (38 cases).Between-group differences were compared in aspects of surgical condition,treatment effect,Visual Analogue Scale (VAS),postoperative hip function (Harris score) and living ability (Barthel index).Postoperative complications were recorded as well.Results Duration of follow-up was 12-24 months [(20.8 ± 1.9) months].Incision length in PFNA group was (5.4 ± 1.2) cm versus (7.3 ±1.7) cm in Gamma nail group (P ＜ 0.01).Operative time in PFNA group was (96.8 ± 24.7) min versus (153.5 ±30.4)min in Gamma nail group (P ＜0.01).Blood loss in PFNA group was (151.5 ±16.8) ml versus (184.7 ± 22.7) ml in Gamma nail group (P ＜ 0.01).Healing time of proximal femur fractures and femoral shaft fractures in PFNA group [(16.1 ± 2.5) weeks,(19.5 ± 3.4) weeks] were significantly less than that in Gamma nail group [(18.9 ± 2.6) weeks,(22.6 ± 2.6) weeks] (P ＜0.01).VAS was (3.4 ±0.5) points in PFNA group versus (6.2± 1.3) points in Gamma nail group (P ＜ 0.01).At postoperative 2 weeks,6 months and 12 months,Harris score in PFNA group [(62.4 ±4.9),(73.7 ± 6.3),(86.4 ± 3.7) points] were significantly higher than that in Gamma nail group [(50.1 ± 4.7),(65.5 ± 6.1),(81.0 ± 3.5) points] (P ＜ 0.01).At postoperative 6 months and 12 months,Barthel index in PFNA group (76.4 ± 6.3,88.1 ± 7.0) were significantly higher than that in Gamma group (70.4 ±5.4,81.3 ±6.9) (P ＜0.01).No implant failure,screw cut-out,coxa vara or fracture displacement occurred.One patient (3％) in Gamma nail group had surgical wound infection compared to PFNA group (P ＞ 0.05).Conclusion In treatment of proximal femoral fractures combined with ipsilateral femoral shaft fractures,the lengthened PFNA fixation has advantages over Gamma nail in terms of minor trauma,rapid postoperative recovery and significant effect.

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 discuss the changes of serum Th1/Th2 cytokine levels of allergic rhinitis patients and the intervention effect of montelukast.Methods 40 cases of allergic rhinitis patients were selected as treatment group,who were given montelukast tablets (10mg) through the mouth one time daily for 8 weeks.The changes of interferon-γ (IFN-γ) and interleukin-4 (IL-4) in peripheral blood (PB) before and after medical treatment were observed,as well as the clinical curative effect and security.Accordingly 30 cases of healthy people were also selected as the control group.Results The IFN-γlevel in culture supernatant of patients in the treatment group [(8.74 ± 1.37) ng/mL] was lower than that in the control group before the medical treatment [(15.13 ± 3.16)ng/mL],while the IL-4 level of patients in the treatment group [(1.64 ± 0.47) ng/mL] was higher than that in the control group [(0.72 ± 0.21) ng/mL] (t =2..93,3.52,all P ＜ 0.01).After 8 weeks' medical treatment,the IFN-γ level in culture supernatant of patients [(12.36 ± 2.56) ng/mL] increased obviously than that of before [(8.74 ± 1.37) ng/mL],while the IL-4 level [(1.21 ± 0.28) ng/mL] declined obviously than that of before [(1.64 ± 0.47) ng/mL] (t =2.36,2.31,all P ＜ 0.05).The total clinical efficiency of patients in the treatment group was 92.5 ％ (37/40).No serious drug adverse reaction (DAR) appeared during the medical treatment.Conclusion The Th1/Th2 cytokine unbalances in peripheral blood (PB) of allergic rhinitis patients.Montelukast has reliable curative effect with high security,whose mechanism of action has close effect on lowering the IL-4 in peripheral blood (PB),increasing the IFN-γ level in peripheral blood (PB),correcting the balance disorders of Th1/Th2 cytokine,and the transformation of immunity reaction from Th2 to Th1.

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 evaluate denervation changes [fibrillation potentials (fib) and positive sharp waves (psw)] in thoracic paraspinal muscles as diagnostic aids in amyotrophic lateral sclerosis (ALS). Methods The fib and psw of thoracic paraspinal muscles,the motor unit potential of sternocleidomastoid muscle were detected in 50 patients who had been identified as having ALS.Age and sex-matched,30 normal subjects and 30 patients who suffered radicular disease in cervical or lumbosacral region were also detected in thoracic paraspinal muscles for finding fib and psw serving as controls. Results In ALS group,a vast of fib and psw were found in thoracic paraspinal muscles in 41 of 50 ALS patients (82%),they were not found with sternocleidomastoid muscle,but the motor unit potential of this muscle was shown having long-duration and high-amplitude in 48 of 50 ALS patients (96%),which were coincided with the chronic neurogenic changes.6 of 50 patients had denervation changes of tongue muscles,3 of them found fib and psw.2 of 30 patients who suffered from radicular diseases were found with a few of fib and psw changes in thoracic paraspinal muscles.None of these symptoms were found in normal subjects.Conclusion A vast of fib and psw in thoracic paraspinal muscles should represent a sensitive tool for diagnosis of ALS.

Objective To measure the difference of radiosensitivity between small and large intestines toward high dose of radiation and investigate the role of stem cells in this difference.Methods C57BL/6 male mice,6-8 weeks old,were randomly divided as control group and radiation group received 19 Gy whole body γ-ray irridiation.Large and small intestines of the mice were collected 6,12,24,48,72 and 96 h after radiation.The proliferation and apoptosis of the large and small intestines and their stem cells were then detected by immunochemistry,and the change of stem cell number in the large and small intestines were detected by in-situ hybridization.Results HE staining showed that 19 Gy γ-ray irradiation caused more severe injury in the small intestine,and all the crypt in the small intestine were extinct at 48 h post-radiation.However,the proliferation index of crypt in the large intestine was as high as 0.23 (t =4.67,P ＜0.05).Compared with the small intestine,the apoptotic index of epithelial cells in the crypt of large intestine was much lower at 12 and 24 h after irradiation (t =-1.92,-2.42,P＜0.05).The apoptotic population of stem cells in the small intestine at 12 and 24 h post irradiation were significantly lower than that in the large intestine (t =-1.98,-2.33,P ＜ 0.05),and the number of stem cell in the large intestine was significantly higher than that in the small intestine 24,48 h after radiation (t =1.98,3.31,P ＜0.05).Conclusions The radiosensitivity of small intestine toward high dose of irradiation is significantly higher than that of the large intestine,where the difference in radiosensitivity of stem cells between large intestine and small intestine may be involved.