The safety of hEPO gene therapy was assessed. No env gene of retrovirus was detected in the EPO-transgenic myoblasts and fibroblasts by PCR detection. Malignant transformation of transgenic cells was ruled out by various examinations including cell morphology, chromosome karyotype, soft-agar test, nude mice test and pathology. All the assessments primarily demonstrated that EPO gene therapy mediated by retrovirus was safe.

Objective To explore the application of bumetanide to inhibition of tumor cell proliferation.Methods In different cell lines, the expression of natrium,kalium, chloride cotransporter 1 ( NKCC1) was detected by Western blotting while the proliferation of different tumor cells was examined by CCK-8 kit.Results The target protein NKCC1 expression in lung cancer cell line ( A549 ) and colorectal cancer cell line ( HCT116 ) was significantly higher than that in chronic myelogenous leukemia cell line (K562), esophageal cancer cell line (Eca109), cervical carcinoma cell line (HeLa), T lymphocytic leukemia cell line (Jurkat) and breast cancer cell line (MCF7).IC50 Values of bumetanide were significantly lower in A549 and HCT116 than in K562, Eca109,HeLa,Jurkat and MCF7.Furthermore, the inhibiory rate and the target protein expression level were positively correlated.Conclusion Bumetanide can inhibit tumor cell proliferation and NKCC1 can serve as a potential target of anticancer drugs.

Objective To evaluate the influence of successful revascularization by percutaneous coronary intervention(PCI)on heart function of patients with heart dysfunction combined with chronic total occlusion(CTO).Methods The clinical data of 272 patients with heart dysfunction combined with CTO were analyzed.The patients were divided into PCI success group(246 cases)and PCI failure group(26 cases)respectively according to the results of PCI.Six months after PCI,the patients underwent cardiac ultrasound examination to compare the heart function between the two groups.Results Cardiac ultrasound examination was successfully performed in 229 patients in PCI success group and 24 patients in PCI failure group at 6 months after PCI.The left ventricular ejection fraction(LVEF)and left ventricular end-diastolic volume index(LVEDVI)showed no significant difference in PCI failure group at 6 months after PCI compared with that before PCI(P＞0.05).In PCI success group,LVEF and LVEDVI were significantly increased at 6 months after PCI compared with that before PCI and compared with that in PCI failure group at 6 months after PCI[(51±5)％ vs.(43±6)％ and(45±2)％,(77±13)ml/m2 vs.(86±12)ml/m2 and(86±10)ml/m2,P＜0.05].The cardiac functional grading in PCI failure group had no significant difference compared with that before PCI(P＞0.05),but in PCI success group it had significant difference compared with that before PCI and compared with that in PCI failure group at 6 months after PCI(P＜0.05).Conclusion Successful revascularization by PCI can improve heart function in patients with heart dysfunction combined with CTO.

Objective Nrf2 is an important neuroprotective factor and the ubiquitin proteasome system ( UPS) , as a highly specific intracellular protein degradation pathway, plays an important role in maintaining gene and protein functions.This paper pres-ents a preliminary study on the relationship between Nrf2 and the ubiquitin proteasome system in the mouse model of traumatic brain in-jury ( TBI) . Methods Forty-two healthy male ICR mice were randomly divided into three groups: control, TBI +sulforaphane ( SFN) and TBI+vehicle, and 12 Nrf2-knockout mice were included in the TBI+Nrf2 -/-group.The animals of the TBI+SFN group were treated with SFN while those of the TBI+vehicle group with the same volume of 10%corn oil at 5 minutes after TBI.At 24 hours after TBI, brain samples were collected from the mice for determining the Nrf2 expression and ubiquitinated protein content by Western blot and the changes in the Nrf2 and ubiquitinated proteins were observed by immunohistochemistry and electron microscopy. Results Compared with the controls, the mice in the TBI+vehicle group showed significantly increased expressions of Nrf2 ( 0.09 ± 0.02 vs 0.66 ±0.09, P<0.05) and ubiquitinated proteins (3.27 ± 0.21 vs 10.58 ±0.75, P<0.05).In comparison with animals in the TBI+vehicle group, those in the TBI+SFN group exhibited a signifi-cant increase in the Nrf2 protein level (0.66 ±0.09 vs 1.22 ±0.14, P<0.05) but a decrease in the ubiquitinated protein level (10.58 ±0.75 vs 6.97 ±0.86, P<0.05), and those in the TBI+Nrf2 -/-group showed a markedly decreased expression of the Nrf2 protein (0.66 ±0.09 vs 0.17 ±0.02, P<0.05) but increased expression of the ubiquitinated protein (10.58 ±0.75 vs 14.35 ± 0.65, P<0.05).Similar results were observed by immunohistochemistry and electron microscopy. Conclusion Nrf2 played a neu-roprotective role in the mouse model of traumatic brain injury by regulating the ubiquitin proteasome system.

AIM: To study a novel gene that probably related with liver regeneration, which was found by representational difference analysis(RDA). METHODS: cDNA sequence, tissue distribution and functions of the novel gene were studied by slot blot, Northern blot, RT-PCR, cDNA library screening and sequence analyzing. RESULTS: Two full-length clones were isolated from cDNA library of rat fetal livers and the sequence analysis identified that the positive cDNA encoded 76 amino acids only; Using the cDNA as a probe, the novel gene showed a specific liver distribution, a moment increasing expression in one hour after partial hepatectomy (PH) and high expression in fetal liver or liver tumor by Northern blot; EGF quickly induced its high expression in primary culture rat hepatocytes(FCS free).CONCLUSION: These results show that the novel gene is an early phase response gene that is closely related to a liver regeneration adjustment. It may encode peptide or has longer sequence at N tip.

Objective To compare the prognosis of vascular in situ and bridge vessel percutaneous coronary intervention ( PCI) therapy strategies in patients with recurrent angina after coronary artery bypass grafting ( CABG) . Methods A total of one hundred and two patients with recurrent angina after CABG from January 2008 to January 2016 were involved in this study and were divided into two groups according to interventional therapy strategy:74 patients in the vascular in situ PCI group ( in situ group,74 cases) and 28 patients for bridge vessel PCI group ( bridge vessel group,28 cases) . The patients have been followed up for (33. 6± 10. 2) months. The major adverse cardiovascular events ( MACE) of the two groups were recorded, including non?fatal acute myocardial infarction ( AMI) ,target vessel revascularization ( TVR) and cardiac death, and multivariate logistic regression analysis was used to analyze the related factors of MACE. Results Compared with the bridge vessel group,the non?MACE survival rate,non?AMI survival rate and non?TVR survival rate of the in situ group were significantly increased ( ( 71. 6% ( 53/74 ) vs. 57. 1% ( 16/28 ) , 93. 2% ( 69/74 ) vs. 82. 1% (23/28),81. 1% (60/74) vs. 67. 9% (19/28) ),the differences were statistically significant (χ2=8. 141,4. 219,5. 436, P<0. 05) . Multivariable logistic regression analysis showed that age of bridge ( OR=1. 023,95%CI 1. 005-1. 026,P=0. 019) ,diabetes mellitus ( OR=2. 386,95%CI 1. 425-3. 991,P=0. 003) and bridge vessel PCI (OR=1. 884,95%CI 1. 093-3. 220,P=0. 025) were factors that affect the clinical prognosis in patients with recurrent angina pectoris after CABG. Conclusion The clinical prognosis of the in situ PCI is better than bridge vascular PCI in patients with recurrent angina after CABG,while the age of bridge, diabetes mellitus, vascular interventional treatment are factors for the effect of interventional therapy patients prognosis. The clinical prognosis is much better in native vessel PCI than that of bridge vessel PCI in patients with recurrent angina after CABG. The age of bridge,diabetes mellitus and bridge vessel PCI are the factors that affect the clinical prognosis in the patients.

Objective Subarachnoid hemorrhage ( SAH) is a devastating disease with high fatality and morbidity and micro-glia/macrophage ( M/M) plays a vital role in SAH brain injury with complicated pathophysiological mechanism .This study was to ob-serve the effect of Nrf2 deficiency on M/M activation and M1 polarization after subarachnoid hemorrhage in mice . Meth ods We col-lected 70 wild-type ( WT) ICR mice and 35 Nrf2-knockout ( KO) mice to establish the SAH model by injecting fresh autologous blood into pre-chiasmatic cistern.WT mice were arranged into four groups: sham operation group, post operative day 1 (POD1) group, POD3 group and POD5 group.Then WT mice and Nrf2 Nrf2-knockout mice were divided into sham operation WT group , sham opera-tion KO group, SAH WT group and SAH KO group.Western blotting (WB) and immunohistochemistry (IHC) were applied to observe the activation and proliferation of M/M after SAH on WT mice .Difference in activation and M 1 polarization were observed by detecting Iba1 expression in WB and CD 16/32 +Iba1 +cells in immunofluorescence between WT and KO mice . Results Gray scale values of Iba1 expression by WB in WT mice are 0.491 ±0.039, 0.657 ± 0.069, 0.930 ±0.046 and 0.926 ±0.046;average optical intensity values of Iba1 expression by IHC in WT mice are 0.412 ±0.122, 0.625 ±0.135, 0.963 ±0.213 and 0.978 ±0.224.The data indica-ted that Iba1 expression increased in SAH KO group in comparison to SAH WT group on 1, 3, 5 day after SAH (P<0.05).Moreover, Nrf2 deficiency promoted the activation and polarization of M /M by increased Iba1 protein expression and CD16/32 +Iba1 +cells after SAH ( P<0.05). Conclusion SAH induces M/M activation and proliferation in mice, and Nrf2 deficiency promotes the activa-tion, proliferation and M1 polarization after SAH .

Objective The prognosis of traumatic brain injury is closely associated with the apoptosis of neural cells .This study investigated the anti-apoptosis effect of alpha lipoic acid (α-LA) and its possible mechanism in the mouse model of traumatic brain injury. Methods Seventy-two healthy male ICR mice were randomly divided into four groups of 18 each:sham operation +vehicle, sham operation +α-LA, trauma +vehicle, and trauma +α-LA.The model of traumatic brain injury was made by weight-dropping.The animals in the α-LA groups were treated with intragastric α-LA at 30 minutes after surgery, while those in the vehicle groups with oral dimethyl sulfoxide in corn oil .At 48 hours after treatment , brain samples were collected from the mice for determining brain edema , measuring the expressions of cytochrome c , Bcl-2-associated X protein ( Bax ) , and caspase-3 in the mitochondria and cytosol of the brain tissue by Western blot and immunohistochemistry respectively , and detecting the survival of the neurons and apoptosis of neural cells in the cortical area by Nissl staining and TUNEL , re-spectively. Results The brain water volume , caspase-3 expres-sion, and neural cell apoptosis were markedly higher while the neuron survival remarkably lower in the trauma +vehicle group than in the sham operation +vehicle and sham operation +α-LA groups ( P<0.01).Compared with the mice in the trauma +vehicle group, those in the trauma +α-LA group showed significantly reduced proportion of water in the brain tissue ([79.89 ±0.55] vs [81.71 ± 0.66]%, P<0.05), expression of caspase-3 ([58.40 ±7.31] vs [47.42 ±7.74]%, P<0.05), and apoptosis of neural cells ([59.63 ±8.61] vs [44.86 ±7.32]%, P <0.05), but increased survival rate of neurons ([44.45 ±10.56] vs [57.46 ± 11.01]%, P<0.05).The expression of cytochrome c in the mitochondria was remarkably decreased and that of Bax markedly in -creased in the trauma +vehicle than in the sham operation +vehicle and sham operation +α-LA groups (P<0.01). Conclusion Alpha lipoic acid has a protective effect against traumatic brain injury in mice , probably by inhibiting the apoptosis of neural cells through the mitochondrial pathway .

Objective To compare the clinical characters and hemodynamic data of the prone position and supine position percutaneous nephrolithotomy. Methods Patients with kidney stones in 100 cases were randomly assigned into the supine position group and prone position group.There were 50 cases in each group.The following data were recorded at preoperative,intraoperative,change position:the blood pressure,heart rate,respiration,saturation of blood oxygen and blood gas analysis results.The VAS score was recorded in patients for postural comfort,dyspnea and pain score. Results The systolic blood pressure in the preoperative,intraoperative,postoperative on supine position and prone position were as follows:(137±12),(119±15),(115±17) mm Hg and (137±10),(110±18),(104±16) mmHg.The diastalic blood pressure was as follows:( 81 ± 9 ),(74 ± 8 ),(63 ± 14 ) mm Hg and ( 84 ± 8 ),(63 ± 9 ),(60 ± 15) mm Hg.Compared with preoperative,there was a blood pressure decreased in both groups.The blood pH at preoperative,intraoperative,postoperative in supine position and prone position:7.4 ± 0,7.3 ± 0,7.3 ± 0 and 7.4 ± 0,7.3 ± 0,7.3 ± 0.The bass excess (BE) of two groups were:(2.1 ± 0.5),(-2.7 ±0.5),( -1.5 ±0.5) mmol/L and (3.2 ±0.5),(-3.8 ±0.5),(-2.5 ±0.5) mmol/L.Compared with preoperative data,the pH and BE declined in both groups.The prone position had more pronounced decrease in serum sodium,serum potassium; but with no significant difference.Supine group in postural comfort and breathing difficulties were 1.6 ± 0.4 and 7.1 4± 05,while prone group were 7.5 ± 0.2 and 1.4 ± 0.3.The supine position showed better in postural comfort and breathing difficulties with statistical significance. Conclusions Compared with the prone position,the supine positioned percutaneous nephrolithotomy could have less influence on hemodynamics,blood gas analysis.It could have better postural comfort.

Objective To investigate the choice of surgical procedures in the treatment of temporal occipital epidural hematomas.Methods From March 2006 to March 2011,176 cases with acute temporal occipital epidural hematomas were treated in our hospital.Their clinical data including preoperative Glasgow Coma Sale (GCS),pupil size,hematoma volume,cerebrospinal fluid leakage,time between injury and operation,cerebral midline shift on CT,and brain beat and brain swelling in the operation were retrospectively analyzed.Results There were significant differences in the choice of surgical treatment and prognosis of temporal occipital epidural hematoma according to the preoperative GCS score,pupillary changes,hematoma volume,length of time before surgery,shift of cerebral midline structures,and brain beat and brain swelling in the operation.Conclusion Appropriate surgical procedures selected according to their preoperative and intraoperative conditions is of significant importance for sound prognosis of the patients with acute temporal occipital epidural hematoma.