Objective To study the changes of gene and protein expressions of key components of renin-angiotensin system (RAS) in myocardium of simulated weightless rats. Method Tail-suspended rats were adopted as animal model of simulated weightlessness. RT-PCR and Western blot analysis were performed to examine mRNA and protein expressions in rat myocardium, respectively. Result After 1 week tail-suspension,no obvious changes in angiotensinogen (AGT), angiotensin converting enzyme (ACE), 1a and 1b subtype of type 1 angiotensin Ⅱ receptor (AT 1aand AT 1b), key components of RAS in myocardium were observed. But after 4 weeks tail-suspension, the expressions of AGT mRNA and higher molecular weight band of AGT protein increased significantly (P

Objective To explore changes of local renin-angiotensin system(RAS)in renal tissue of rat during short-and mid-term tail suspension,as well as their relations with humoral regulation and renal function induced by real or simulated weightlessness.Methods 1-and 4-week(wk)tail-suspended rat model were used to simulate short-and mid-term weightlessness,respectively.Reverse transcriptase polymerase chain reaction(RT-PCR)was carried out to examine the mRNA expression of components of local RAS in renal tissue.Results Except for angiotensin Ⅱ receptor type 2(AT2),all of the other components of local RAS,including angiotensinogen(AGT),renin,angiotensin converting enzyme(ACE),angiotensin Ⅱ receptor type 1a(AT1a)and angiotensin Ⅱ receptor type 1b(AT1b),were found their expression in renal tissue of Sprague-Dawley rats.After 1 wk of tail suspension,mRNA expression of renin in renal tissue increased significantly(P

Objective To investigate the efficiency and safety of ultrasound-mediated microbubble destruction in enhancing β-galactosidase gene (β-gal gene) transfer into human proximal tubular cells(HKCs). Methods β-gal gene was transfected to HKCs as a mark gene with ultrasound-mediated microbubble destruction. Cultured HKCs were grouped to receive the following 7 treatments respectively: ultrasound alone; microbubble alone; naked plasmid; ultrasound and plasmid; microbubble and plasmid; ultrasound, microbubble, and plasmid; and VigoFect and plasmid. In Group 6, HKCs were exposed to ultrasound under different sound intensities and time. X-gal stainning, typan blue stainning, and Hochest stainning were used to detect the transfection efficiency, cell survival rate, and cell apoptosis rate, respectively.Results β-galactosidase expression could be observed in the ultrasound-mediated microbubble destruction groups. Along with the increasing of sound intensity and exposure time, the cell survival rate of HKCs decreased, and the cell apoptosis rate increased gradually. The transduction efficiency and survival rate in middle intensity (0.3 W/cm~2×60 s) of ultrasound exposure were higher than those of other groups, similar to those of Group 7.Conclusion Under optimum sound intensity and exposure time, ultrasound-mediated microbubble destruction can increase gene transfer into HKCs. This non-invasive gene transfer method may be a useful tool for clinical gene therapy.

Objective To explore the diagnostic consistency and correlation between MR discography (MRD) and CT discography (CTD) in diagnosing chronic low back pain. Methods Guided by C - arm fluoroscopy the mixed solution of gadoterate meglumine (GD-DOTA) and Iohexol (GD-DOTA at a dilution of 1 ∶ 400 with Iohexol) was injected into 96 lumbar intervertebral discs of the 36 patients. CT scanning was performed at 15 minutes after the injection of contrast, and axial together with sagittal SE T1WI MR scanning was carried out one hour after the injection. CTD and MRD images were randomly numbered and were independently evaluated by two experienced radiologists according to Dallas discogram scale in order to assess the diagnostic consistency and correlation between (MRD) and (CTD). In addition the diagnostic value of MRD was evaluated. Results The results revealed that in determining disc degeneration grade CTD and MRD were highly consistent with each other(Kappa = 0.836, P < 0.01), and the diagnostic results judged by the two reviewers were essentially in agreement (ICC = 1.00, P < 0.01; r = 0.997, P < 0.01). Higher consistency (Kappa = 0.836, P < 0.01) and correlation(ICC = 0.90, P < 0.01; r = 0.869, P < 0.01; Kappa =0.836, P < 0.01) in determining annulus rupture extent were also obtained. Conclusion MRD is an accurate diagnostic method for the determination of disc degeneration and the severity of annulus rupture, and this technique has greater consistency and correlation with CTD in diagnosing chronic low back pain.

ObjectiveTo investigate the changes in mammalian target of rapamyein (mTOR) with aging in rat kidneys.MethodsMale Wistar rats at the ages of 3, 12, 24 months were used for this study. Therenaltissuesandmesangialcellswereprocessedfor senescenceassociated β-galactosidase (SA-β-gal) staining. The expression and location of roTOR in kidneys and mesangial cells were studied by immunohistochemistry and immunocytochemistry, respectively. The mRNA and protein levels of the roTOR and p-roTOR were detected by Western blot assay and RT-PCR,respectively.ResultsThe expression of neutral β-galactosidase activity was increased in kidneys and mesangial cells with advancing age. Percentages of SA-β-gal staining positive ceils were (11.9±3.6)% versus ( 39.0±4.0)% versus ( 86.9±7.4) % in young, middle and aging glomerular mesangial cells (P<0.05). The mTOR staining appeared in the mesangial matrix and interstitium in kidneys, while the mTOR protein showed localization in cytoplasm and nucleus in mesangial cells. The staining intensity of mTOR in kidneys and mesangial cells in aged rats was markedly increased as compared to that in young and middle aged rats (P<0.05). The mRNA level of roTOR was significantly increased in kidneys and mesangial cells of agedrats versus young and middle aged rats,meanwhile, the roTOR and p-mTOR protein expressions were dramatically increased with advancing age (P<0.05 ).ConclusionsmTOR expression is increased with aging, which may play an important role in the aging process of kidneys.

Objective To explore the mechanism of SH?SY5Y mitochondrial dysfunction treated by manganese to find a new potential therapeutic target. Methods Transmission Electron Microscopy(TEM)to observe the morphology of mitochondria. Cell treated with 250μmol/L for periods of time(2 h, 4 h, 6 h)while mitochondrial membrane potential(MMP)and ROS can be detected by FCM and fluorescence microplate reader. Results After treating with MnCl2 in 6 h, TEM images showed early vacuoles, lamellar structures of SH?SY5Y cells. Then test the mitochondrial membrane potential and showed that MMP would be decreased gradually. Meanwhile, analysis showed that in comparison with control, treatment group had a higher ROS level respectively (P < 0.05). Conclusion MnCl2 can cause mitochondrial damage through a mechanism closely related to disrupt the MMP or generate abundant ROS.

Objective To investigate the clinical efficacy of short segment fixation with percutaneous pedicle screws or traditional open surgery for the type A1-A3 thoracolumbar compression fracture.Methods A retrospective case control study was conducted on the clinical data of 64 patients with thoracolumbar compression fracture admitted to Shanxi Dayi Hospital between January 2012 and February 2017.There were 44 males and 20 females,aged 21-65 years [(45.4 ± 11.1) years].There was one patient with injured segment at T11,29 at T12,27 at L1 and seven at L2.According to AO typing,there were 39 patients classified as Type A1,two as Type A2 and 23 as Type A3.The patients were divided into minimally invasive surgery group (n =37) and open surgery group (n =27).Minimally invasive surgery group was treated with minimally invasive percutaneous pedicle screw fixation and open reduction.The open surgery group was treated with traditional open pedicle screw short segment fixation and open reduction.The operation time,intraoperative blood loss,total hospitalization time,postoperative hospitalization time,visual analogue scale (VAS) before and after operation,local kyphosis of the fractured vertebra,segmental kyphosis and complications in two groups were recorded.Results All patients were followed up for 12-29 months,with an average of 13.2 months.Between the minimally invasive surgery group and open surgery group,no significant difference was found in the operation time [(106.4± 37.3) minutes vs.(131.3 ± 33.6) minutes] (P ＞ 0.05),and significant differences were found in intraoperative blood loss [(71.2 ± 34.9) ml vs.(409.3 ± 267.5) ml],total hospitalization time [(11.7 ± 7.2) days vs.(21.6 ± 12.8) days] and postoperative hospitalization time [(8.1 ± 7.4) days vs.(16.6 ± 10.6) days] (P ＜ 0.05).In the minimally invasive surgery group,VAS was (6.5 ±1.1) points preoperatively and was (2.3 ± 0.7) points and (1.0 ± 0.3) points immediately after operation and at final follow-up.In the open surgery group,VAS was (6.9 ± 1.0)points preoperatively and was (4.2 ± 1.0) points and (0.9 ± 0.4) points immediately after operation and at final follow-up (P ＜0.05).Compared with the preoperative VAS,those immediately after operation and at final follow-up were significantly decreased within the two groups (P ＜ 0.05).There were no significant differences in the preoperative VAS and VAS at final follow-up between the two groups (P ＞ 0.05),but significant difference was found in VAS immediately after operation between the two groups (P ＜ 0.05).In the minimally invasive surgery group,the local kyphosis of the fractured vertebra was (19.3 ± 3.8) °preoperatively,(3.4 ± 1.7) ° immediately after operation,and (4.6 ± 1.9) ° at final follow-up.In the open surgery group,the local kyphosis of the fractured vertebra was (19.6 ± 6.8) ° before operation,(1.6 ± 0.8) ° immediately after operation,and (2.4 ± 1.1) ° at final follow-up.The kyphosis of fractured vertebra immediately after operation and at final follow-up were significantly decreased within the two groups compared with the preoperative kyphosis(P ＜0.05),but no significant differences were found between the two groups (P ＞ 0.05).In the minimally invasive surgery group,the segmental kyphosis Cobb angle was (16.1 ± 9.1) ° before operation,(3.0-± 1.8) ° immediately after operation,and (5.9 ±1.8) ° at final follow-up.In the open surgery group,the segmental kyphosis Cobb angle was (15.2±12.0) ° before operation,(3.1 ± 1.4) ° immediately after operation,and (5.6 ± 2.1) ° at final follow-up.The segmental kyphosis Cobb angle immediately after operation and at final follow-up were significantly decreased within the two groups compared with the preoperative Cobb angle (P ＜ 0.05),but no significant differences were found between the two groups (P ＞ 0.05).No spinal cord injuries because of pedicle screws were observed after operation in either group.In the open surgery group,there was one patient with wound infection who recovered after dressing change,and no infection case was found in the minimally invasive surgery group.Conclusion For type A1-A3 thoracolumbar compression fractures,both the minimally invasive posterior pedicle screw fixation and the traditional open pedicle screw fixation can achieve satisfactory near-term results,and the former is better in intraoperative blood loss,immediate relief of pain after operation and shorter hospital stay than the latter.

OBJECTIVETo investigate the role of tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) in regulating both angiogenesis and the expressions of phosphatase and tensin homologue deleted on chromosome 10 (PTEN) and vascular endothelial growth factor (VEGF)/Flk-1 expression in human proximal tubular epithelial cells (HKC).

METHODSHKC cells were transfected with two recombinant plasmids containing sense and antisense full-length TIMP-1 cDNA (TIMP-1S-pcDNA3.0 and TIMP-1AS-pcDNA3.0, respectively) constructed previously, or treated with 100 µmol/L MMP-2/MMP-9 inhibitor III (with similar cellular enzyme suppression activity with sense TIMP-1 plasmid). The mRNA expression of TIMP-1, MMP-2, MMP-9, PTEN, VEGF and Flk-1 were examined by RT-PCR. In each group, the expression of PTEN, VEGF and Flk-1 were also detected using an indirect immunofluorescence assay.

RESULTSCompared with non-transfected cells and cells transfected with the empty vector, sense TIMP-1-transfected cells showed obviously upregulated PTEN expression (P<0.05) and significantly lowered gelatinase activity (P<0.05) and VEGF and Flk-1 expressions (P<0.05). Transfection with the antisense TIMP-1 plasmid produced the reverse results (P<0.05). MMP-2/MMP-9 inhibitor III did not obviously affected the expression of PTEN, VEGF or Flk-1 as compared with the non-transfected or empty vector-transfected cells.

CONCLUSIONIn the aging progress, the renal tissues express high levels of TIMP-1 to upregulate PTEN expression via a MMP-independent pathway, and subsequently down-regulates the expression of VEGF and Flk-1 to cause aging-related impairment of renal angiogenesis. These findings provide new evidence for understanding the role of TIMP-1 in renal aging.

Cells, Cultured ; Epithelial Cells ; metabolism ; Humans ; Kidney Tubules, Proximal ; cytology ; Matrix Metalloproteinase Inhibitors ; PTEN Phosphohydrolase ; metabolism ; RNA, Messenger ; genetics ; Tissue Inhibitor of Metalloproteinase-1 ; genetics ; metabolism ; Transfection ; Vascular Endothelial Growth Factor A ; metabolism ; Vascular Endothelial Growth Factor Receptor-2 ; metabolism

Mild cognitive impairment (MCI), as a nosological entity referring to elderly people with MCI but without dementia, was proposed as a warning signal of dementia occurrence and a novel therapeutic target. MCI clinical criteria and diagnostic procedure from the MCI Working Group of the European Alzheimer's Disease Consortium (EADC) may better reflect the heterogeneity of MCI syndrome. Beijing United Study Group on MCI funded by the Capital Foundation of Medical Developments (CFMD) proposed the guiding principles of clinical research on MCI. The diagnostic methods include clinical, neuropsychological, functional, neuroimaging and genetic measures. The diagnostic procedure includes three stages. Firstly, MCI syndrome must be defined, which should correspond to: (1) cognitive complaints coming from the patients or their families; (2) reporting of a relative decline in cognitive functioning during the past year by the patient or informant; (3) cognitive disorders evidenced by clinical evaluation; (4) activities of daily living preserved and complex instrumental functions either intact or minimally impaired; and (5) absence of dementia. Secondly, subtypes of MCI have to be recognized as amnestic MCI (aMCI), single non-memory MCI (snmMCI) and multiple-domains MCI (mdMCI). Finally, the subtype causes could be identified commonly as Alzheimer disease (AD), vascular dementia (VaD), and other degenerative diseases such as frontal-temporal dementia (FTD), Lewy body disease (LBD), semantic dementia (SM), as well as trauma, infection, toxicity and nutrition deficiency. The recommended special tests include serum vitamin B12 and folic acid, plasma insulin, insulin-degrading enzyme, Abeta40, Abeta42, inflammatory factors. Computed tomography (or preferentially magnetic resonance imaging, when available) is mandatory. As measurable therapeutic outcomes, the primary outcome should be the probability of progression to dementia, the secondary outcomes should be cognition and function, and the supplement outcome should be the syndrome defined by traditional Chinese medicine. And for APOE epsilon4 carrier, influence of the carrier status on progression rate to dementia and the effect of treatment should be evaluated.

In order to provide the "guiding principles of clinical research on mild cognitive impairment (MCI) (protocol)" edited by Beijing United Study Group on MCI of the Capital Foundation of Medical Developments (CFMD) with evidence support, clinical criteria, subtypes, inclusion and exclusion of MCI, and use of rating scales were reviewed. The authors suggested that MCI clinical criteria and new diagnosis procedure from the MCI Working Group of the European Alzheimer's disease Consortium (EADC) may better reflect the heterogeneity of MCI syndrome. Diagnostic rating scales including Clinical Dementia Rating (CDR), Global Deterioration Scale (GDS), Alzheimer's Disease Assessment Scale-cognitive subscale (ADAS-cog) and Instrumental Activities of Daily Living (IADL) are very useful in definition of MCI but can not replace its clinical criteria. Absence of major repercussions on daily life in patients with MCI was emphasized, but the patients may have minimal impairment in complex IADL. According to their previous research, the authors concluded that highly recommendable neuropsychological scales with cut-off scores in the screening of MCI cases should include Mini-Mental State Examination (MMSE), logistic memory test such as Delayed Story Recall (DSR), executive function test such as Clock Draw Test (CDT), language test such as Verbal Category Fluency Test (VCFT), etc. And finally, the detection of biological and neuroimaging changes, including atrophy in hippocampus or medial temporal lobe in patients with MCI, was introduced.