Objective To investigate the effect of preconditioning with different doses of emulsified isoflurane on focal cerebral ischemia-reperfusion (I/R) injury in rats. Methods Ninety-six male SD rats were randomly divided into 6 groups ( n = 16 each): sham operation group (group S), I/R group, low, median and high doses of emulsified isoflurane preconditioning group (group L, M, H) and intralipid group (group IL). Middle cerebral artery occlusion was produced by inserting a nylon thread. In group S, intraperitoneal normal saline (NS)10.5 ml/kg was injected, but the artery was only exposed 24 h later. In group I/R, intraperitoneal NS 10.5 ml/kg was injected, and the model was established 24 h later. In group L, M, H and IL, 8% emulsified isoflurane 3.5 ml/kg+NS 7.0 ml/kg, 8% emulsified isoflurane 7.0 ml/kg + NS 3.5 ml/kg, 8% emulsified isoflurane 10.5 ml/kg and 30% intralipid 10.5 ml/kg were injected intraperitoneally respectively, and then the model was established 24 h later. The temperature, HR and RR were recorded at 10 min before ischemia and at 10 min of reperfusion. The neurological deficit was scored, the cerebral infarct volume and apoptosis in neurons were detected, and microscopic examination of ischemic penumbra region was performed at 24 h of reperfusion. Results The rectal temperature and HR were significantly increased, while RR was significantly decreased duing cerebral I/R.The neurological deficit score was significantly higher, cerebral infarct volume and the number of apoptotic neurons were significantly larger in the other groups than in group S. Emulsified isoflurane preconditioning reduced neurological deficit scores, cerebral infarct volume and the number of apoptotic neurons dose-dependently. Conclusion Emulsified isoflurane preconditioning can reduce focal cerebral I/R injury in a dose-dependent manner in rats.

Objective To investigate the effect of postconditioning with 8 % emulsified isoflurane (EI) on focal cerebral ischemia-reperfusion (I/R) injury in rats. Methods Forty-eight male SD rats weighing 260-300 g were randomly divided into 6 groups ( n = 8 each): group Ⅰ sham operation (group S); group Ⅱ I/R; group Ⅲ,Ⅳ, Ⅴ 3 different doses of EI (group L-EI, M-EI, H-.EI) and group Ⅵ lipid emulsion (group LE). Right middle cerebral artery occlusion (MCAO) was induced by inserting a nylon thread 0.24 mm in diameter into right internal carotid artery. The thread was threaded cranially until resistance was met. MCAO was maintained for 2 h followed by 24 h reperfusion in group Ⅱ-Ⅵ. In group Ⅲ-Ⅴ EI 3.5, 7.0 and 10.5 ml/kg were injected intraperitoneally (IP) immediately before reperfusion respectively. In group LE 30% lipid emulsion 10.5 ml/kg was given instead of EI. The neurologic deficit was assessed and scored (0 = normal, 4 = unable to move and unconscious). The animals were then killed and infarct size was measured. Results IP 8% emulsified isoflurane 7.0 and 10.5 ml/kg injected before reperfusion significantly reduced neurologic deficit scores and infarct size in group M-EI and H-EI as compared with group I/R. Conclusion Postconditioning with 8% emulsified isoflurane can protect the brain against focal cerebral I/R injury.

Objective To investigate the effect of preconditioning with emulsified isoflurane (eISO) on neuronal apoptosis in hippocampal CA1 region induced by focal cerebral ischemia-reperfusion (I/R) injury in rats. Methods Forty-eight healthy adult male SD rats weighing 250-300 g were randomly divided into 6 groups (n = 8 each): sham operation group (group S); I/R group; eISO + I/R group (group EI); LY294002 (a specific PI3K inhibitor) + eISO + I/R group (group L+ EI); LY294002 + I/R group (group L) and DMSO (solvent for LY294002) + I/R group (group DMSO). Focal cerebral I/R was induced by 2 h middle cerebral artery occlusion ( MCAO). A nylon thread (0.26 mm in diameter) with rounded tip was inserted into internal carotid artery and advanced cranially until resistance was met (depth of insertion about 18-20 mm) . eISO 10.5 ml/kg (120 mg/ml) was injected intraperitoneally (IP) in groups EI and L+ EI. LY294002 (25 mmol/L) 5 pi was injected into cerebral ventricle on the ischemic side in group L + EI ( at 30 min before eISO) and group L. DMSO 5 μl was injected into the cerebral ventricle on ischemic side before MCAO in group DMSO. Neurologic deficit was assessed and scored (0 = normal, 4 = unconscious) at 24 h of reperfusion. The animals were then killed and their brains were removed for detection of neuronal apoptosis (by TUNEL) and p-Akt expression (by immuno-histochemistry) in hippocampal CA1 region. Results Cerebral I/R significantly increased the neurologic deficit scores, the number of apoptotic cells and p-Akt expression in group I/R as compared with group S. Preconditioning with elSO attenuated the I/R-induced increase in neurologic deficit scores and number of apoptotic cells but further increased p-Akt expression. The neuroprotective effect of eISO preconditioning against I/R-induced changes was counteracted by LY294002. Conclusion eISO preconditioning can attenuate focal cerebral I/R-induced neuronal apoptosis in rats by activating PI3K/Akt pathway.

Objective To investigate the changes of bone mineral density (BMD) in human immunodeficiency virus (HIV)-infected patients in Changsha,and take intervention measures to prevent the occurrence of osteoporosis and fracture.Methods A total of 278 HIV-infected patients and 154 cases of healthy adults from March 2011 to May 2015 were selected.Dual energy X-ray absorptiometry (DXA) was used to detect BMD,T-score and Z-score of all the research objects,including the whole body,lumbar spine (L2～4),and left hip joint.The height and weight were measured at the same time.Results The HIV infection group had an average age of (31.53 ± 8.56) years old,and the healthy control group was (34.45 ± 8.22) years old.Height between two groups had no significant difference.The average weight of HIV infection group was 6.93 kg [95％ CI,-9.01,-4.97;P ＜0.001] lighter than that in the normal control group.BMD,T-score and Z-score of HIV infection group were significantly lower than those in norrmal control group (P ＜ 0.001).The occurrence rate of osteopenia (Z ≤-1.0)and osteoporosis (Z ≤-2.0)in HIV infection group were correspondingly 43.53％ ～ 54.68％ and 9.71％ ～23.74％,which is about 4 times of that in the healthy control group (14.28％ ～ 20.13％,0.65％ ～ 5.84％).Conclusions The average body weight of HIV-infected patients was significantly lower than that of normal control group,and the incidence of osteopenia and osteoporosis in HIV-infected group was significantly higher than that in normal control group.

Objective On the basis of internet of things technology,to initially establish a management model of chronic obstructive pulmonary disease (COPD) patients in Gumei community,so as to provide experience for the comprehensive management of community COPD patients.Methods According to the characteristics of the Internet of things technology,we formulated a scheme as a technical route to manage the COPD patients.A homogenous COPD management team of doctors was established under the training of experts from the Department of Respiration of Zhongshan Hospital,Fudan University.Results We drew a COPD patient management model chart,and initially formed a qualified and homogeneous COPD management team of general practitioner.Conclusions Through the Internet of things technology management,we initially formed a set of manual quality control model in the process of data automatic transmission,and initially formed a management model of community COPD patients,based on the internet of things.

This study was aimed to explore the features of clustered regularly interspaced short palindromic repeats (CRISPR) structures in Shigella by using bioinformatics. We used bioinformatics methods, including BLAST, alignment and RNA structure prediction, to analyze the CRISPR structures of Shigella genomes. The results showed that the CRISPRs existed in the four groups of Shigella, and the flanking sequences of upstream CRISPRs could be classified into the same group with those of the downstream. We also found some relatively conserved palindromic motifs in the leader sequences. Repeat sequences had the same group with corresponding flanking sequences, and could be classified into two different types by their RNA secondary structures, which contain "stem" and "ring". Some spacers were found to homologize with part sequences of plasmids or phages. The study indicated that there were correlations between repeat sequences and flanking sequences, and the repeats might act as a kind of recognition mechanism to mediate the interaction between foreign genetic elements and Cas proteins.
Base Sequence ; Clustered Regularly Interspaced Short Palindromic Repeats ; Computational Biology ; Genome, Bacterial ; Plasmids ; Shigella ; genetics

Aim To investigate the expression of stro-mal interaction molecule 1 (STIM1) in rat pulmonary arterial hypertension ( PAH ) tissues and effects of STIM1 on arterial muscle cells proliferation. Methods PAH was induced by a single intraperitoneal injec-tion of MCT at a dose of 60 mg·kg - 1 . The mRNA or protein expressions of STIM1 in monocrotaline-induced pulmonary hypertensive rats were measured by real-time PCR or Western blot, respectively. The arterial smooth muscle cells A7R5 were transiently transfected with STIM1 plasmids to prepare STIM1 overexpressed cells. Cell proliferations were detected by using CCK-8 kits. The expressions of Akt/ mTOR pathway molecules of A7R5 were measured by Western blot. Results The right ventricular systolic blood pressure ( RVSP) and right ventricular mass index ( RVMI ) were markedly elevated in MCT-treated rats (P < 0. 01) in comparison to control rats. The mRNA and protein ex-pression levels of STIM1 in monocrotaline-induced pul-monary hypertensive rats were 2. 19 and 1. 66 folds of control rats, respectively. STIM1 were transiently over-expressed in cultured A7R5. Cells transfected with STIM1 grew more quickly than non-transfected control. Overexpression of STIM1 significantly increased the phosphorylation of Akt, mTOR, p70-S6K, and 4E-BP1, but did not change their protein expression lev-els. Conclusion STIM1 are over-expressed in rat PAH tissues. Overexpression of STIM1 can promote ar-terial smooth muscle cells proliferation by regulating Akt/ mTOR pathway.

Aim To investigate the effect of store-oper-ated calcium channel( SOCC) on autophagy in rat arte-rial smooth muscle cells A7 R5 . Methods Lentiviruses containing STIM1 or Orai1 gene were packaged in 293 T cells and then were used to infect rat arterial smooth muscle cells A7 R5 . The expression levels of STIM1 , Orai1 and Beclin 1 , a critical autophagy-regu-lating protein, of lentivirus-infected A7R5 cells, were detected by Western-blot. Autophagy in lentivirus-in-fected A7 R5 cells was induced by starvation or rapamy-cin, an inhibitor of mammalian target of rapamycin ( mTOR ) . Autophagy marker LC3 of these cells was detected by Western-blot. Results The constructions of vector pLV-STIM1 and pLV-Orai1 were confirmed by restriction enzymes digestion analysis. Compared with the control group, expressions of STIM1 or Orai1 protein was significantly increased after lentivirusLV-STIM1 and LV-Orai1infection, whereas the expressions of autophagy related protein Beclin-1 were down-regu-lated. Starvation or rapamycin stimulated A7R5 auto-phagy but overexpression of STIM1 or Orai1 significant-ly inhibited starvation or rapamycin induced autoph-agy. Conclusion Overexpression of store-operated calcium channel components STIM1 and/or Orai1 in rat arterial smooth muscle cells A7 R5 inhibit autoph-agy. This mechanism might contribute to the develop-ment of pulmonary arterial hypertension.

Objective To evaluated the clinical value of laparoscopic techniques in radical cystectomy surgery for the treatment of bladder cancer. Methods Clinical data of 49 patients underwent radical cystectomy with Bricker ileal conduit diversion were retrospectively analyzed from October 2009 to August 2014, which laparoscopic radical cystectomy with Bricker ileal conduit 20 cases (Group A), open radical cystectomy with Bricker ileal conduit 29 cas-es (Group B). The blood loss during operation, operating time, gastrointestinal function recovery after operation, hos-pital stay after operation and complications were observed between the two groups. Results The blood loss during operation was significantly lower in Group A (416.66 ± 232.73) ml than in Group B (964.16 ± 445.73) ml ( <0.05), and hospital stay after operation was significantly lower in Group A (14.93 ± 2.72) days than in Group B (19.50 ± 3.16) days ( < 0.05), complication after operation was significantly lower in Group A than in Group B ( < 0.05). The operating time and gastrointestinal function recovery has no significantly difference between the two groups. Conclusions Laparoscopic radical cystectomy have advantages of minimal invasion, less blood loss, rapid recovery and less postoperative complications. It is a safe and effective surgical method. Long term effect need evaluated by follow up.