Aim To construct the eukaryotic expression vectors of short hairpin RNA targeting survivin and observe its effect on biologic behavior of A549 cells and sensitivity of A549 cells to paclitaxel.Methods The DNA fragment targeting human survivin was inserted into the plasmid,and the recombinant plasmid was constructed.The recombinant plasmids cells were transfected into A549 cells by FuGENE transfection reagent.The expression levels of survivin gene were detected with RT-PCR and Western blot before and after transfection,respectively.Cell apoptosis was detected by TUNEL method.The sensitivity of A549 cells to paclitaxel was detected by MTT after transfection.Results The recombinant plasmid was successfully constructed.RNAi group cells showed lower expression of survivin than control group.The apoptosis rate of A549 cells increased after transfection.The IC50 of paclitaxel inhibiting A549 cells was 11.9 fold before transfection compared with those after transfection.There was significant difference between the two groups(P

Objective To establish a semi-quantitative method for measurement of HCV RNA by use of primers and probe, which was sensitive and designed by ourselves, a new europium fluorescent chelate BHHCT. Methods 44 serum samples of HCV infected patients and 20 samples of the healthy people were collected. HCV RNA in serum sample was extracted by HCV fluorescence PCR diagnostic kit produced by Zhongshan University DAAN Gene Co Ltd, and amplified by RT-PCR in which one PCR primer was pre-labeled with biotin. The amplified products were hybridized with capture probes pre-fixed on the microplate. The biotin in the amplified products was conjugated with europium labeled streptavidin. So europium was linked to the target DNA. Then the fluorescent of europium was measured. Results The linear range of this assay was 10 - 10 copies/ml. Both sensitivity and specificity were 100%. Conclusion Europium labeled RT-PCR assay is a sensitive, specific, fast and non-radioactive contaminant method for the measurement of HCV RNA.

OBJECTIVE To investigate the nosocomial infection knowledge nurses in graduating class. METHODS 286 nurses of graduating class were investigated by questionnaires. RESULTS The students were lacked of knowledge about nosocomial infection and 61.70% nurses of graduating class had never participated in the training of nosocomial infection knowledge (the average score only 58.67). They were interested in the disinfection and sterilization of infection diseases. And they wanted to get it through clinical infection control group (81.56% students selected this educational methods). CONCLUSIONS It is necessary to strengthen the training and education of nosocomial infection. And adjust the training mode and content according object.

BACKGROUND:Titanium mesh has good clinical effect in repairing skul defects, but due to the lack of bone induction ability, the titanium mesh has a poor integration with the bone tissue. OBJECTIVE:To observe the biological properties of the gradient bioactive coating materials on the titanium surface in the skul repair. METHODS: Osteoblasts were co-cultured with the titanium mesh with or without gradient bioactive coatings for 14 days, and then cel proliferation was detected using MTT method. Seventy-one patients with skul defects were enroled, including 43 males and 28 females, aged 15-60 years, and were subjected to skul repair using the titanium mesh with (observation group, n=3) or without (control group, n=38) gradient bioactive coatings. During the postoperative folow-up of 12 months, the repairing effects and adverse reactions were observed in the two groups. RESULTS AND CONCLUSION:(1)In vitrocel culture: the cel proliferative ability was increased significantly in the observation group as compared with the control group at 8, 10, 12 and 14 days after cel culture. (2)In vivo repair: the hospital stay and wound healing time in the observation group were significantly lower than those in the control group (P< 0.05), and at the final folow-up, the postoperative recovery effect was significantly higher in the observation group than the control group (P < 0.05). The titanium meshes were fixed firmly in the two groups, with no floating, infection and exposure. These results show that the titanium mesh with gradient bioactive coating has good biocompatibility and osteoinduction capacity.

Objective Study the endobronchial ultrasound features of mediastinal tubercular lymphadenopathy and sarcoidosis to probe a new method for the differential diagnosis. Methods The endobronchial ultrasound features of 74 lymph nodes in 16 mediastinal tubercular lymphadenopathy and 30 sarcoidosis patients diagnosed in our department were studied retrospectively, and the sizes, borders, fusion and echo features of mediastinal tubercular lymphadenopathy were compared to sarcoidosis. Results Both of the long size and the short size of mediastinal tubercular lymphadenopathy were smaller than sarcoidosis [(15.77 ± 4.10) vs (19.76 ± 5.83), t = 3.28, P = 0.021;(12.67 ± 4.09) vs (16.81 ± 5.54), t = 3.56, P = 0.001]. And the following features were statistically significant of tubercular lymphadenopathy as compared to sarcodosis: indistinct borders, fusion of lymph nodes, hyperechoic echotexture and patchy anechoic/hypoechoic areas [50.0% (11/22) vs 17.3% (9/52), χ2 = 8.38, P = 0.004; 18.2%(4/22) vs 0.0% (0/52), P = 0.008; 50.0% (11/22) vs 0.0% (0/52), P = 0.000; 63.6% (14/22) vs 0.0% (0/52), P = 0.000, respectively). However, there was no significant difference in the existence of central hilar structure [9.1% (2/22) vs 19.2% (10/52), P = 0.491] between mediastinal tubercular lymphadenopathy and sarcoidosis. Conclusions The endobronchial ultrasound features of mediastinal lymph nodes, including sizes, borders, fusion, hyperechoic echotexture and patchy anechoic/hypoechoic areas are helpful in the differential diagnosis of mediastinal tubercular lymphadenopathy and sarcoidosis.

Objective To evaluate left atrial(LA)function in patients with prophase essential hypertension (EH)by left atrial volume tracking(LAVT)method.Methods Thirty prophase EH patients and 35 healthy subjects(control group)were enrolled in this study.Ultrasound LAVT(Hitachi EUB-6500)was applied to display and analyze the LA volume loop imaging on the standard LV apical two and four chamber views.The maximal LA volume at end-systole(LAVmax),LA volume at the onset of ECG-P wave(LAVP),the minimal LA volume at end-diastole(LAVmin)from the LA volume loop were recorded,and body surface area was used to revise these volume indexs(LAVImax,LAVIp,LAVImin).LA pass,act and total empting volume(LAVIpass,LAVIact,LAVItotal)and empting rate(%LAVIpass,%LAVIact,%LAVItotal),effective pass and act erupting rate(%eLAVIpass,%eLAVIaet),and the proportionality of pass empting volume and act empting volume were caculated.Results The LAVIp,LAVIact,LAVItotal,%LAVIact,%LAVItotal,%eLAVIact in the prophase EH group were significantly higher than those in the control group,wheras the LAVIpass,%LAVIpass,%eLAVIpass,LAVIpass/act were lower(all P＜0.05,or P＜0.01).Conclusions The LA volume change in prophase EH is the act empting volume increased at end-diastole in main,LAVT is a potientially useful tool to evaluate the function of LA.

The value of the left atrial volume tracking (LAVT) method in the evaluation of left atrial (LA) function in patients with diabetes mellitus (DM) was examined in this study. Fifty-eight DM patients as DM group and 40 healthy people as normal control group were enrolled in this study. EUB-6500 echocardiographic imaging system with LAVT was applied to display and analyze the LA volume curve imaging on LV apical two and four chamber views. The maximal LA volume at end-systole (LAV(max)), LA volume at the onset of ECG-P wave (LAV(p)), the minimal LA volume at end-diastole (LAV(min)) from the LA volume curve were acquired and recorded. All values above were standardized by body surface area (BSA). Then the passive, active and total LA volume (LAVIpass, LAVIact, LAVItotal) and empting rate (%LAVIpass, %LAVIact, %LAVItotal), effective passive and active empting rate (%eLAVIpass, %eLAVIact), and the proportionality of passive empting volume and active empting volume were calculated. The LAVIp, LAVIact, LAVItotal, %LAVIact, %LAVItotal and %eLAVIact were significantly higher in the DM group than those in the control group, whereas the LAVIpass, %LAVIpass, %eLAVIpass and LAVIpass/act were lower (all P<0.05). For the LA volume change in DM, the active empting volume was enhanced at end-diastole. It was concluded that LAVT is a potentially useful tool to evaluate the function of LA.

Objective To investigate the expression of RECK gene in placentas of patients with preeclampsia and its correlation with MMP-2 activation,and explore the possible roles of RECK gene in the placental trophoblast invasion mechanism.Methods RT-PCR and Western blot were used to detect the expression of RECK mRNA and protein respectively in the placental tissues of normal late pregnant women (normal pregnant group,22 cases) and pre-eclamptic patients(22 mild cases and 20 severe cases).Gelatinase zymography was used to determine MMP-2 activation ratio.Results The expression levels of RECK mRNA and protein from placenta tissues in mild,severe pre-eclamptic group were both significantly higher than those in nomal pregnant group.Moreover,the expression levels of RECK mRNA and protein in severe pre-eclamptic group were obviously increased as compared with those in mild pre-eclamptic group.There was significant difference among the three groups (all P<0.01).MMP-2 activation ratio in mild,severe pre-eclamptic group was significantly lower than that in normal pregnant group.MMP-2 activation ratio in severe pre-eclamptie groups was obviously reduced as compared with mild pre-eclamptic group.There was significant difference among the three groups(all P<0.01).The expression leVels of RECK mRNA and protein were significantly negatively correlated with MMP-2 activation ratio (both P<0.01).Conclusion The abnormal high expression of RECK and inhibition of MMP-2 activation in placentas of pre-eclamptic patients may participate in the process of placental trophoblast shallow invasion.

The changes in the tau protein phosphorylation and expression of bcl-2, and bax in rat parietal cortex neurons after focal cerebral ischemia-reperfusion (I/R) were explored, and the relationship between the tau protein phosphorylation and the expression of bax or apoptosis was clarified in order to elucidate the relationship between cerebral infarction and Alzheimer's disease. The rat focal cerebral I/R model was induced by occlusion of the right middle cerebral artery using the intraluminal suture method. The level of tau protein phosphorylation at Ser396, Ser404, Tyr231, Ser199/202 sites and the expression of bcl-2, bax and total tau 5 in rat parietal cortex during focal cerebral ischemia/reperfusion were detected by Western blot. The relationship between the tau protein phosphorylation and the expression of bax, or apoptosis was examined by TUNEL method and double-labeling immunofluorenscence method. The results showed that the level of tau hyperphosphorylation at Ser199 / 202, Ser396, Ser404, Tyr231 sites and the expression levels of bcl-2, and bax were significantly higher in I/R group than in the sham group, but the ratio of bcl-2/bax was decreased. Neuronal apoptosis, bax expression and the tau protein hyperphosphorylation were co-localized. It is suggested that Alzheimer's disease-like pathological changes occur after cerebral I/R. The highly abnormal phosphorylation of tau protein plays a key role in cerebral I/R-induced apoptosis. The cerebral infarction may contribute to Alzheimer's disease occurrence and development.

Transthoracic echocardiographic characteristics of 17 cases of cardiac amyloidosis (CA), a rare disease in China, were analyzed in order to improve the understanding of the disease. Seventeen cases of biopsy-proven CA, admitted to Wuhan Union Hospital from June 1994 to September 2008 were retrospectively reviewed. Twenty normal volunteers served as control group. Left atrial and ventricular functions and mitral inflow velocity were measured by two-dimensional, and Doppler echocardiography, and tissue Doppler imaging (TDI)-derived peak systolic wall motion velocities (Sv), peak early diastolic wall motion velocities (Ev), and peak late diastolic wall motion (Av) were measured at the septum, lateral, inferior and anterior corners of mitral annulus from the apical 4- and 2 chamber views. Compared with the control group, the interventricular septal thickness (IVSd), the left ventricular posterior wall (LVPWd), right ventricular transverse diameter (RVTDd) near the end of diastole and the interauricular septum thickness (IASs), left atrial anteroposterior diameter (LAADs), right atrial transverse diameter (RATDs) near the end of systole were increased significantly (all P<0.05) and left ventricular ejection fraction (LVEF) decreased (P<0.05) in the CA group. Compared with the control group, Sv, Ev at each wall and Av at almost all walls were significantly decreased in the CA group. In the CA group, Myocardial echoes of interventricular septum and free wall of left ventricle were enhanced evidently and distributed unevenly. The echoes presented as ground glass-like images, with some spotty hyper echoes. Both atria were enlarged, and LVEF decreased, with diastolic function impaired, and mild-moderate hydropericardium found in the CA group. It was concluded that echocardiography was a relatively sensitive and highly specific non-invasive method for the diagnosis of CA.
Amyloidosis/*ultrasonography ; Cardiomyopathies/*ultrasonography ; Case-Control Studies ; Echocardiography ; Echocardiography, Doppler/*methods ; Retrospective Studies ; Sensitivity and Specificity