BACKGROUND: Hippocampus injury is wildly believed to involve in neurocognitive dysfunction; the establishment of a rat model of cardiopulmonary bypass(CPB) allows us to investigate the mechanism of CPB-related hippocampus injury.OBJECTIVE: To investigate the effects of moderate hypothermic CPB with a hemodilution on hippocampal bcl-2 and bax gene expression and neuronal apoptosis in rats.DESIGN: A randomized group division study based on the experimental animals.SETTING: Department of anesthesiology in a university hospital.MATERIALS: Thirty Sprague-Dawley (SD) rats were randomly divided into two groups, CPB group and sham-CPB group with 15 rats in each group.METHODS: Total 15 rats of CPB group were subjected to 60-minute moderate hypothermic nonpulsatile CPB using a peristaltic pump and a membrane oxygenator. The CPB circuit was primed with approximately 20 mL 1:1crystaloid-colloid liquid, while another 15 rats of sham-CPB group underwent identical anesthetic and surgical procedures(including cannulation) except CPB itself. At 1 hour post-CPB, six rats in each group were decapitated, and hippocampi were removed, homogenized, and processed for apoptotic gene ( bcl-2 and bax) mRNAs detection. Reverse transcriptase polymerase chain reaction(RT-PCR) is used to detect expression of mRNA by comparing the PCR product of bcl-2 or bax to those of β-actin housekeeping gene. Immunohistochemistry is used to detect bcl-2 and bax protein expressions and terminal deoxynucleiotidyl transferase-mediated dUTP-biotion nick end labeling(TUNEL) staining method was used to detect neuronal apoptosis at 6 hours post-CPB ( n = 6 in each group) . The protein expression was quantitated as percentage of the positively stained area in the total stained. In addition, hippocampal neuronal ultrastructures were studied by electron microscopy at 6 hours post-CPB( n = 3 in each group).ronal apoptosis and ultrastructure changes between the two groups.RESULTS: At 1 hour post-CPB, the expressions of bcl-2 and bax gene, and the ratio of bax to bcl-2 in CPB group were significantly increased compared with those of sham-CPB group( P ＜ 0.05) . At 6 hours post-CPB, the expressions of Bcl-2 and Bax protein in hippocampal CA1 region had significantly increased in CPB group(P ＜ 0.05) . TUNEL staining showed that hippocampal CA1 neuronal apoptosis was significantly increased in CPB group compared with sham-CPB group at 6 hours post-CPB( P ＜ 0. 05).Electron Microscopy demostrated that, at 6 hours post-CPB, neuronal ultrastructures in CPB group had obvious abnormalities, many mitochondria being moderately to severely swollen with vacuolation as well as decreasing number of mitochondrial cristae, some neurons having characteristic morphological changes of earlier periods of apoptosis such as neuronal pycnosis, irregular nucleus, nuclear membrane with notchs, chromatin condensation, and nucleoli movement to the nuclear periphery, etc.CONCLUSION: Moderate hypothermic CPB with a hemodilution can induce hippocampal bax, bcl-2 gene expression and neuronal apoptosis in rats,which maybe partly explain the mechanism of post-CPB neurocognitive dysfunction.

Objective To investigate the effects of mild hypothermic cardiopulmonary bypass(CPB) on bax and bcl-2 protein expression and neuronal apoptosis in hippooampus CA1 region in rats. Methods Healthy male SD rats weighing 380-420g were randomly divided into 3 groups: (1)post-CPB 1h group (n=6); (2) post-CPB 6h group (n=6) and (3) sham CPB group(n=5). The animals were anesthetized with intraperitoneal fentanyl 150?g?kg~(-1), midazolam 1 mg?kg~(-1) and vecurunium 0.1 mg?kg~(-1), intubated and mechanically ventilated. Left femoral artery was cannulated for heparinizafion, BP monitoring and blood sampling. Artery in the tail and right jugular vein were cannulated for CPB. The animals in group 1 and 2 underwent 2 h mild hypothermic (26-28℃) CPB at a flow of 160 ml?kg~(-1)?min~(-1). 1h(group 1) and 6h(group 2) after CPB 4% polymerized formaldehyde was injected via the artery in the tail. Brains were immediately removed. Bax and bcl-2 protein expression in hippocampus CA1 region was determined by immuno-histochemistry method and neuronal apoptosis by TUNEL technique. Ultrastructural changes were examined with electron microscope. Results (1)Bax and Bcl-2 protein expressions were significantly higher in group 1(1h after CPB)than in sham CPB group. Bax protein expression was significantly higher but bcl-2 protein expression was significantly lower in group 2 than in group 1. bax/bcl-2 ratio in group 2(at 6h after CPB)was significantly higher than that in group 1(at 1h after CPB). (2) Neuronal apoptosis was significantly increased in group 1 and 2 compared to sham CPB group. (3) Electron microscopy showed that some mitochondria were moderately or severely swollen with vacuolizafion and decreased number of mitochondrial at 1h after CPB and at 6h after CPB there was shrinkage of neuronal cells with irregular shape of nucleus, notches in nuclear membrane and margination of nucleoli. Conclusion CPB induces upregulation of both bax and bcl-2 protein expression with predominance of the pro-apoptotic gene Bax leading to increased neuronal apoptosis, indicating that CPB can cause brain injury.

Objective To investigate the relationship between cerebral 0, metabolism during coronary artery bypass graft (CABG) performed with cardiopulmonary bypass (CPB) and postoperative mental disorders (POMD) in the elderly.Methods Thirty patients (13 male,17 female) aged 65-76 yr undergoing CABG performed with CPB were studied. Patients with senile dementia were excluded. The patients were premedicated with scopolamine 0.3 mg and pethidine 50mg. Anesthesia was induced with midazolam 0.05-0.1 mg?kg_(-1), propofol 0.2-0.5 mg?g_(-1), fentanyl 10-15?g?g_(-1) and vecuronium 0.1-0.3 mg?g_(-1) and maintained with isoflurane inhalation and intermittent iv boluses of fentanyl. Nasopharyngeal temperature (T) was maintained at 26℃-28℃) during CPB. Radial artery was cannulated for BP monitoring and blood sampling.Swan-Ganz was placed via right internal jugular vein (UV).Another catheter was inserted into right UV and threaded retrogradely until J point for blood sampling. Blood samples were taken from artery and UV simultaneously for blood gas analyses and determination of blood glucose and lactate concentration before anesthesia (T_1 ),immediately after induction of anesthesia (T_2), during splitting of sternum (T_3) when T was reduced to 28℃ (T_4) and rewarmed to 35℃(T_5 ) and at the end of bypass (T_6 ).Cerebral O2 delivery (CDO2 ),cerebral oxygen consumption (CMRO2),cerebral glucose consumption (CMRglu) and lactic acid production (ADVL) were calculated. POMD was assessed using CAM-ICU chart.Results (1)Seven patients developed POMD (7/30 or 23.3% ).(2) Low jugular bulb 02 saturation (SjO2 ≤50% or PjO2≤25% mm Hg) was observed in 8 patients during rewarming. The incidence was significantly higher in POMD patients (3/7 or 42.96%) than that in non-POMD patients (5/23 or 21.74%). (3) CBF/CMRO2 was significantly lower in POMD patients than in non-POMD patients during rewarming.(4) CMRO2/CMRglu was significantly lower but ADVL was significantly higher in POMD patients than in non-POMD patients during hypothermia, rewarming and at the end of CPB.(P

Objective To evaluate the role of NF-κB in astrocytes in the spinal dorsal horn in a rat model of neuropathic pain induced by paclitaxel.Methods Twenty 6 year old male SD rats weighing 180-200 g were randomly divided into 2 groups ( n =10):control group (group C) and neuropathic pain group (group NP).Neuropathic pain was induced by intraperitoneal injection paclitaxel of 2 mg/kg every other day for 4 times in group NP.While the equal volume of normal saline was given instead of paclitaxel in group C.Body weight,mechanical pain threshold and heat pain threshold were determined before and at 1,7,14 d after administration.The animals were sacrificed and the spinal dorsal horn were removed for determination of NF-κB p65 expression in astrecytes by immunofluorescence analysis at 14 d after administration.Results There was no significant difference in body weight between the two groups( P ＞ 0.05).Compared with group C,mechanical pain threshold and heat pain threshold were significantly decreased 7 and 14 d after administration,the expression of NF-κB p65 in astrocytes in the spinal dorsal horn was up-regulated in group NP( P ＜ 0.05 or 0.01 ).Conclusion Activation of NF-κB in astrocytes in the spinal dorsal horn may play an important role in neuropathic pain induced by paclitaxel in rats.

Objective To learn the clinical features of severe pneumonia in northeast sichuan ,and to improve the diagnosis and treatment level .Methods Basic disease or concomitant diseases and it′s clinical features were retrospective analyzed in different age patients with severe pneumonia in 2011 .Results (1)basic diseases and fatality cases /deteriorated cases were increased in elderly patients with the increase of age .There were 4 patients infected with HIV (3 cases under 65 years old ,1 case over 70 year old) .(2) onset pattern was given first place to suffer from cold ,but 40% were showed for symptoms of nervous system ,heart disease and other respiratory system ;(3)The main respiratory failure types of severe pneumonia is Ⅰ (about 70% ) ,30% of patients needed support of machine ventilation ;(4)community pneumonia were main type(75% );detection rate of Fungal infections and G -bacteria were 25% -30% ,normal bacteria (Streptococcus and Neisser bacteria )were over 1/3 ,Staphylococcus aureus were about 10% ;(6) More than 70% of the patients with different degree of hypoalbuminemia ;(7)Cure accounts for 2/3 ,poor prognosis accounted for 1/3 ,in which death and voluntary discharge were each 16 .6% ,ventilator-dependent ,multi-organ failure ,infections and some com-plex social factors were the prognostic factors .Conclusion Understanding the features and treatment difficulty of the local commu-nity acquired pneumonia is helpful to improve the timely diagnosis ,treatment and rescue level of patients with severe pneumonia .

Porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus type 2 (PCV2) are very two important pathogens that have coursed huge economic losses in swine production in worldwide. In this study,a vector pCMV-TJM containing the full-length cDNA clone of PRRSV attenuated strain TJM-F92 was firstly constructed by PCR method. Then a gene sequence containing Afl II/Mlu I e restriction enzyme sites and a transcription regulatory sequence for ORF6 (TRS6) was inserted be- tween ORF7 and 3'UTR, yielding a expression vector pCMV-TJM-TRS. Subsequently, a plasmid pCMV-TJM-Cap was constructed by cloning of PCV2 ORF2 gene into the unique sites Afl II /Mlu I of pCMV- TJM-TRS plasmid DNA. Then three recombinant PRRSV, rTJM, rTJM/TRS and rTJM/Cap, were rescued by transfection of pCMV-TJM, pCMV-TJM-TRS and pCMV-TJM-Cap into Marc-145 cells, respectively,and confirmed by the genome sequence, restriction enzyme digestion, Western Blot and IFA. They all had the molecular markers which was different from the parent virus. The growth characteristics of the rescued viruses were similar to that of parent virus. rTJM/Cap could also express efficiently PCV2 Cap protein in Marc-145 cells. At passage 8, it still had PCV2 ORF2 gene which examined by RT-PCR. It indicated that the full-length cDNA clone of PRRSV attenuated strain TJM-F92 and recombinant PRRSV rTJM/Cap expressing PCV2 Cap protein were successfully constructed. It made an important foundation for studying on the pathogenic mechanisms of PRRSV and PRRSV-PCV2 vaccine in the future.
Animals ; Capsid Proteins ; genetics ; immunology ; Cell Line ; Circoviridae Infections ; veterinary ; virology ; Circovirus ; classification ; genetics ; metabolism ; Gene Expression ; Open Reading Frames ; Porcine Reproductive and Respiratory Syndrome ; virology ; Porcine respiratory and reproductive syndrome virus ; genetics ; metabolism ; Recombination, Genetic ; Swine ; Swine Diseases ; virology ; Viral Vaccines ; genetics ; immunology

AIM: To study the effects of ?-amyloid protein (A?) on neural stem cells cultured in vitro. METHODS: Neural stem cells (NSC) were isolated from E13 SD rats and cultured in serum-free medium (DMEM/F12). After detected by nestin, the A? was added to the NSC medium to observe the viability and proliferation of NSC by MTT, cell count and flow-cytometric examination. The effects of A? on differentiated NSC were also observed. RESULTS: A? markedly inhibited the proliferation and the cell viability of NSC when its concentration was higher than 25 ?mol/L. The differentiatory ability of NSC was inhibited when A? was in very low concentration. CONCLUSION: A? significantly inhibits the proliferation and differentiation of NSC and this may be one of the reasons that Alzheimer's disease is induced. [

AIM: To study the effects of [Gly~(14)]-humanin,one of the strongest derivate of humanin,on proliferation and differentiation of neural stem cells(NSCs) and the protective action of cell death or apoptosis induced by ?-amyloid protein 1-42.METHODS: NSCs were treated with different concentrations of [Gly~(14)]-humanin and ?-amyloid protein 1-42.RESULTS: 10 nmol/L [Gly~(14)]-humanin made NSCs resistant to the apoptotic action induced by A?P1-42 and prevented NSCs from death induced by 25 ?mol/L ?-amyloid protein 1-42.The differentiated neural stem cells yield more neuronal cells than that in control groups when 10 nmol/L [Gly~(14)]-humanin was added to the culture media.The number of cells increased and the cultures grown with a manner of floating cell clones likes that cultured in the presence of mitogen when 100 nmol/L [Gly~(14)]-humanin was added to the differentiation culture media.CONCLUSION: The [Gly~(14)]-humanin significantly promoted the proliferation and neuronal differentiation of neural stem/progenitor cells and also inhibited the toxic action of ?-amyloid protein 1-42 on cultured neural stem/progenitor cells.

Objective: To study the correlation between left atrial sphericity (LASP) and thromboembolic events (TE) in patients with atrial fibrillation (AF). Methods: This study was conducted in patients with AF underwent radiofrequency ablation in the Department of Cardiology of First Affiliated Hospital of Zhengzhou University from January 2011 to October 2018. The AF patients with TE (study group, n=157) and the AF patients without TE (control group, n=157) were matched for age and gender. The differences of LASP and other related indexes between the two groups were compared, and the correlation between LASP and TE was analyzed by conditional logistic regression. The receiver operating characteristic (ROC) curve was drawn to analyze the diagnostic value of LASP for TE. Results: (1) The LASP in the study group was significantly higher than that in the control group [ (87.5±7.1) % vs. (82.8±6.1) %, P=0.001]. (2) Conditional logistic regression analyses showed that LASP (OR=1.10, 95%CI 1.05-1.16, P=0.001), left atrial volume index (OR=1.01, 95%CI 1.00-1.02, P=0.016) and CHA₂D-VASc score (OR=1.77, 95%CI 1.30-2.41, P=0.001) were independently and positively correlated with TE. (3) The ROC curve analysis showed that the area under the curve (AUC) of left atrial sphericity (AUC=0.712, 95%CI 0.656-0.768, P=0.001) was larger than the AUC of either left atrial volume index (AUC=0.650, 95%CI 0.589-0.710, P=0.001) or CHA₂D-VASc score (AUC=0.612, 95%CI 0.550-0.674, P=0.001). (4) CHA₂D-VASc-LASP₂ score was positively correlated with TE (OR=1.95, 95%CI 1.55-2.42, P=0.001). Conclusion LASP is independently and positively correlated with TE in patients with AF.

Objective To examine the efficacy and safety of pregabalin in patients with neuropathic cancer pain (NCP).Methods A prospective randomized control multicenter trial was conducted in five hospitals;from January 2015 to January 2016,one hundred and twenty two eligible inpatients and outpatients were divided into pregablin treatment group (n=60) and control group (n=62).Patients in the pregablin group added pregablin to opiod background analgesia,while those in the control group raised opioid dose instead.The Numerical Pain Rating Scale (NRS) scores,paraesthesia scale scores,Hamilton's Depression (HAMD) scale scores,analgesia dose,patiends satisfaction,and adverse events were recorded 14 d after each treatment.Results After each treatment,the NRS scores were decreased by (2.3 ±1.1) and (1.3±1.5),the paraesthesia scale scores were decreased by (1.6±0.6) and (0.4±0.3),and the HAMD scale scores were decreased by (4.4±1.2) and (2.4±1.0) in the pregablin treatment group and control group,respectively,with significant differences (P＜0.05).Morphine dose for breakthrough pain in pregabalin group was statistically less than that in control group ([30.6±3.5] mg/d vs.[70.9±12.3] mg/d,P＜0.05).Patients satisfaction in the pregablin treatment group was significantly higher than that in the control group (P＜0.05).Pregabalin treatment group had less severe adverse effects (3/56,5％) as compared with control group (10/59,16.1％,P＜0.05).Conclusion Pregabalin has positive roles in patients with NCP already receiving opioid;pregabalin has better pain-control and mood improvement.