Objective To investigate and compare the effects of pretreatment with different concentrations of trarnadol and morphine on the differentiation of hunan helper T cells (Th) in vitro. Methods Fifteen ASA Ⅰ patients of both sexes (8 males, 7 females) aged 23-59 yrs scheduled for minor day-care surgery were enrolled in this study. The patients had never taken morphine and tramadol before. Blood samples were taken from peripheral vein. Whole blood and mononuclear cells isolated from peripheral blood (PBMCs) were incubated with morphine 10, 100 or 1000 ng?ml-1(group M1 , M2 , M3) or tramadol 50, 500, 5000 ng?ml-1(group Q1, Q2, Q3) for 24 h. Then irrntants was added and incubated for another 5 or 24 h. In the whole blood the level of IL-2 and IFN-?(as markers of Th1 cells) and IL-4 and IL-10 (as markers of Th2 cells) were determined using three-color flow cytometry. In PBMCs the expression of CD4+ and CCR5+ (as markers of Thl cells) and CD4+ and CCR3+ (as markers of Th2 cells) were determined. The Th1/Th2 ratio was calculated. Results Pretreatment with morphine or tramadol significantly increased the number of Th2 cells while the number of Thl cells significantly decreased both in a dose-dependent manner leading to a significant decrease in Th1/Th2 ratio. Conclusion Tramadol and morphine can inhabit in vitro the cellular immune function in dose-dependent manner. The immunosuppressive effect of tramadol is less than that of morphine.

Objective To investigate the effects of pretreatment with different concentrations of sufentanil of ketamine on the differentiation of human helper T cells in vitro. Methods Twenty-two healthy volunteers (11 males, 11 females) aged 20-45 yrs were enrolled in this study. In each volunteer 20 ml of blood was taken from peripheral vein and divided into 7 groups: control group (0.9% NaCl), 3 sufentanil groups (0.05, 0.5, 5.0 ng?ml-1) and 3 ketamine groups (100, 500, 2 500 ng?ml-1) .Whole blood and mononuclear cells from peripheral blood (PBMCs) were incubated in the presence of 0.9% NaCl or different concentrations of sufentanil or ketamine for 24 h. Then the stimulants-phorbolmyristate + lonomycin + glgistap (inhibitor of intracellular protein transport) were added to whole blood and phytohemagglutinin (PHA) was add to PBMCs. The whole blood was incubated for another 4h and PBMCs were incubated for another 48 h. Then the T-lymphocytes were collected for determination of intracellular level of IFN-?(as a marker of Th1 cells) and IL-4 (as a marker of Th2 cells) in the whole blood using three-color flow cytometry and the expression of CCR5 + (as a marker of Th1 cells) and CCR3 + (as a marker of Th2 cells) in PBMCs. The Th1/Th2 ratio was calculated. Results In sufentanil 0.5 and 5.0 ng?ml-1 groups the percentage of Th2 cells was significantly increased while the percentage of Th1 cells and Th1/Th2 ratio were significantly decreased. In ketamine 500 and 2 500 ng?ml-1 groups the percentage of both Th1 and Th2 cells were significantly decreased and the Th1/Th2 ratio was significantly increased. Conclusion Sufentanil can encourage helper cells to differentiate into Th2 cells while ketamine inhibit the helper cells to differentiate into Th1 and Th2 cells, especially the Th2 cells in a dose-dependent manner.

Objective To evaluate the effect of minocycline on the long-term cognitive function after partial hepatectomy in mice.Methods Sixty-four male C57BL/6 mice,aged 3 months,weighing about 20 g,were randomly divided into 4 groups (n =16 each) using a random number table:control group (group C),partial hepatectomy group (group O),PBS group,and minocycline group (Mino group).At 3 days after partial hepatectomy,the rats underwent Morris water maze test.The animals were sacrificed and their hippocampi were immediately removed for determination of tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) mRNA (using real-time fluorescent quantitative PCR) and nuclear factor kappa B (NF-κB) expression in the nuclcar protein of the cells (using Western blot).After the mice regained consciousness after operation,minocycline 50 mg· kg-1 · d-1 was injected intraperitoneally for 30 consecutive days in Mino group,while the equal volume of PBS was given instead of minocycline in group PBS.The parameters mentioned above were then recorded.Results Compared with group C,the escape latency and swimming distance were significantly prolonged,and the expression of TNF-α mRNA,IL-6 mRNA and NF-κB was up-regulated in group O (P ＜ 0.05).Compared with group PBS,the escape latency and swimming distance were significantly shortened,and the expression of TNF-α mRNA,IL-6 mRNA and NF-κB was down-regulated in group Mino (P ＜ 0.05).Conclusion Minocycline can improve the long-term cognitive function after partial hepatectomy in mice and inhibition of inflammatory responses in hippocampi is involved in the mechanism.

Objectives To explore the relationship of orthostatic hypotension in elderly hypertensive patients with left ventricular structure and function.Methods 101 cases of elderly patients with hypertension were selected from July 2013 to June 2015.After blood pressure measurement in vertical and decubitus position,all patients were divided into non-orthostatic hypotension and orthostatic hypotension groups,received the echocardiography and the accurate calculation of left ventricular mass index.Results Compared with non-orthostatic hypotension,orthostatic hypotension in patients with left ventricular diastolic diameter,left ventricular posterior wall thinckness and interventricular septum thickness.Left ventricular mass index significantly increased(48.13±5.54)mm vs.(52.45 ± 1.48)mm in left ventricular diastolic diameter,(10.08± 1.87)mm vs.(11.29± 1.98)mm in left ventricular posterior wall thickness,(10.18± 1.88)mm vs.(11.61±1.66) mm in interventricular septum thickness,and (114.59 ± 22.72) g/m2 vs.(131.98 ± 23.43)g/m2 [t=4.386、2.985、4.397、8.672,in left ventricular mass index(BMI),all P＜0.05].Left heart ventricular ejection fraction and mitral peak early/late diastolic blood flow decreased [(60.81 ± 4.73)vs.(53.60±2.58)and(0.93±0.23)vs(0.76±0.26)t=4.298、3.654,all P＜0.05].Conclusions There is relationship of orthostatic hypotension with left ventricular structure and function in elderly hypertension patients,and it is essential not only to control blood pressure effectively,but also to focus on controlling orthostatic hypotension.

BACKGROUND: The spinal instability would accelerate the degeneration of normal disk. The injuries of anterior longitudinal ligament (ALL) and intervertebral disk were usually caused by cervical trauma, which leaded to spinal instability. Currently, there were few animal researches about the degeneration of injured intervertebral disk affected by ALL destruction.OBJECTIVE: To investigate the degeneration of injured intervertebral disk after spinal instability in the rabbit model of different degrees of ALL and disc destruction.METHODS: A total of 24 New-Zealand rabbits were randomly divided into intervertebral disk injury group (Group A, n=6), partial injury of ALL with disc injury group (Group B, n=6), injury of bony attachment point of ALL with disc injury group (Group C, n=6) and entirely injury of ALL with disc injury group (Group D, n=6). The L2-L3 intervertebral disk and ALL were injured through abdominal cavity. Different groups received different treatments. Computed tomography (CT) scans of the injured disc were performed at the postoperative 4 and 8 weeks, and the middle high of injured discs was calculated on CT sagittal reconstruction. Three rabbits were selected from each group. Hematoxylin-eosin staining of injured disc was performed after the animals were killed. Results were examined under the light microscope.RESULTS AND CONCLUSION: (1) At postoperative 4 weeks, the middle height of injured discs in Group D was decreased significantly compared with Group A (P < 0.05). There were hyperosteogeny and calcification in Group C and Group D. There were nucleus pulposus cells reduction and inflammatory reaction in Group C and Group D on histological staining. (2) At postoperative 8 weeks, the middle height of injured discs respective was decreased significantly compared with Group A (P < 0.05). The hyperosteogeny and calcification became clearer in Group C and Group D than before. There were morphologic changes of nucleus pulposus cells and fibrillation by hematoxylin-eosin staining, and the degree of disc degeneration was Group D > Group C > Group B > Group A. (3) In conclusion, the injury of ALL would accelerate the degeneration of correspondingly injured disk, and the degree of injury of ALL was positively correlated with the degeneration of disk.

BACKGROUND:Endogenous hydrogen sulfide can be used as a new gaseous signaling molecule, and has important signal transfer function and biological regulation effects. OBJECTIVE:To study the neuroprotective effects of hydrogen sulfide in rats with acute cauda equina syndrome. METHODS: The 72 Sprague-Dawley rats were randomly assigned to three groups. Experimental group, model group: laminectomy was performed at the lumbar 4 (L4) level of the vertebra, and a piece of silicone (10 mm long, 1 mm thick, and 1 mm wide) was placed under the laminae of the L5-6 vertebra to produce the animal model of cauda equina syndrome. Sham surgery group: a simple laminectomy was performed in L4, but silicone was not implanted. In the experimental group, 20 μmol/kg NaHS was injected intraperitonealy at 1 hour before model establishment. Model and sham surgery groups: an equal volume of saline was injected intraperitonealy. At 12, 24, 48 and 72 hours after model establishment, malonaldehyde and glutathione levels in cauda equina nerve tissue were detected. Simultaneously, hematoxylin-eosin staining and TUNEL staining were performed at 48 hours. RESULTS AND CONCLUSION:Hematoxylin-eosin staining demonstrated that cauda equina nerve tissue was dense and regular, with complete myelin sheath, no axon sweling in the sham surgery group. Cauda equina nerve tissue was sparse, with the presence of demyelination, and partial axons and myelin sheath sweling in the model group. Cauda equina nerve tissue was tight, with axonal sweling and demyelination in the experimental group. TUNEL staining demonstrated that the number of positive cels was less in the spinal cord and dorsal root ganglia in the sham surgery group. Abundant positive cels were detected in the spinal cord and dorsal root ganglia in the model group. The number of positive cels was significantly lower in the experimental group than that in the model group. Malonaldehyde levels were lower in the sham surgery and experimental groups than in the model group (P < 0.05, P < 0.01), but glutathione levels were higher than model group (P < 0.05,P < 0.01). These results indicated that hydrogen sulfide could decrease oxidative stress and protect cauda equina nerve in rats with acute cauda equina syndrome.

Objective To evaluate the role of astrocyte chemokine (C-C motif) ligand 2 (CCL2) in microglial activation in an in vitro experiment.Methods Primary astrocytes and microglias were isolated from the brain tissues of C57BL/6J mice at postnatal day 1-2.The experiment was performed in two parts.Experiment Ⅰ Astrocytes were inoculated in 6-well culture plates at a density of 3 × 104 cells/well (2 ml/well) and divided into 5 groups (n=3 each) using a random number table:control group (group C),tumor necrosis factor-alpha (TNF-cα) group,1 μg/ml CCL2 small interference RNA (siRNA) group (group CCL2-siRNA1),2 μg/ml CCL2-siRNA (group CCL2-siRNA2) and negative control siRNA group (group NC-siRNA).Astrocytes were cultured routiuely in group C,and 10 ng/ml TNF-α was added and astrocytes were incubated for 15 min followed by washout with phosphate buffer solution (PBS),and then astrocytes were incubated for 3 h in the other 4 groups.At 24 h before TNF-α was added,CCL2-siR-NA 1 and 2 μg/ml were added in CCL2-siRNA1 and CCL2-siRNA2 groups,respectively,and NC-siRNA 2 μg/ml was added in group NC-siRNA.The concentrations of CCL2 were determined by enzyme-linked immunosorbent assay.Experiment Ⅱ Microglias were inoculated in 6-well culture plates at a density of 3×104 cells/well (2 ml/well) and divided into 3 groups (n=3 each) using a random number table:control group (group C),TNF-α group and CCL2-siRNA group.Microglias were cultured routinely in group C.In group TNF-α,10 ng/ml TNF-α was added to astrocytes which were incubated for 15 min followed by washout with PBS,astrocytes were then incubated for 3 h,and the supernatant was collected and added to microglias which were incubated for 24 h.In group CCL2-siRNA,2 μg/ml CCL2-siRNA was added to astrocytes which were incubated for 24 h,10 ng/ml TNF-α was also added to astrocytes which were incubated for 15 min followed by washout with PBS,astrocytes were then incubated for 3 h,and the supernatant was collected and added to microglias which were incubated for 24 h.The activity of microglias was measured by immunofluorescence,and the migration of microglias was evaluated by Transwell migration assay.Results Experiment Ⅰ The concentrations of CCL2 were significantly higher in TNF-α,CCL2-siRNA1,CCL2-siRNA2 and NC-siRNA groups than in group C (P＜0.05).The concentrations of CCL2 were significantly lower in CCL2-siRNA1 and CCL2-siRNA2 groups than in TNF-α and NC-siRNA groups (P＜0.05).There was no significant difference in CCL2 concentrations between group TNF-α and group NC-siRNA (P＞0.05).Experiment 1Ⅱ Compared with group C,the activity of microglias was significantly increased,and the migration of microglias was enhanced in TNF-α and CCL2-siRNA groups (P＜0.05).Compared with group TNF-α,the activity of microglias was significantly decreased,and the migration of microglias was weakened in group CCL2-siRNA (P＜0.05).Conclusion Astrocyte CCL2 is involved in mieroglial activation in an in vitro experiment.

BACKGROUND:Under hypoxic environment, hypoxia-inducible factor 1α plays a dualregulatory role in cel apoptosis. Severity of hypoxia is the key to determine whether cels appear to have apoptosis or adapt to survive. When the cels are exposed to chronic or extreme hypoxia, a lack of protection mechanisms from hypoxia-inducible factor-1α can induce cel apoptosis. OBJECTIVE: To research the expression of hypoxia-inducible factor 1α in human lumbar nucleus pulposus of different herniated types and its relationships with cel apoptosis. METHODS: The nucleus pulposus was harvested from 60 cases of herniation of lumbar intervertebral discs, L4-5 in 41 cases and L5-S1 in 19 cases. The nucleus pulposus tissues were equaly divided into protruded and sequestered groups. Meanwhile, the nucleus pulposus tissues from another 10 cases of lumbar spine fracture were taken as control group. Expression of hypoxia-inducible factor 1α and apoptosis of lumbar nucleus pulposus cels were observed and detected with immunohistochemical technique and TUNEL method. Correlation of hypoxia-inducible factor 1α and apoptosis in human lumbar nucleus pulposus of different herniated types was analyzed. RESULTS AND CONCLUSION: The expression of hypoxia-inducible factor 1α was visualized in each case, but it was significantly higher in the sequestered group than in the protruded group and control group (P < 0.01). Apoptosis of nucleus pulposus cels were found in al the three groups, but the apoptotic rate was also higher in the sequestered group than in the protruded group and control group (P < 0.01). Expression of hypoxia-inducible factor 1α was positively correlated with cel apoptosis in human lumbar nucleus pulposus (P < 0.01). Overal, the expression of hypoxia-inducible factor1α in degenerative human lumbar nucleus pulposus is associated with herniated types, which is the highest in the sequestered type. The relationship between hypoxia-inducible factor 1α and apoptosis is positive.

BACKGROUND:Under hypoxic environment, hypoxia inducible factor-1 plays an important role in regulation of hypoxia-induced gene expression in the intervertebral disc. Hypoxia-inducible factor-1 consists of α and βsubunits, and which hypoxia inducible factor-1α determines the stability and activity of hypoxia-inducible factor-1. OBJECTIVE: To observe the expression of hypoxia inducible factor-1α in the human lumbar nucleus pulposus of different herniated types and to judge their relationships. METHODS:A total of 60 nucleus pulposus samples were harvested from the lumbar vertebra, including 41 from L4-5 and 19 from L5-S1, and then divided into protruded group and sequestered group, with 30 cases in each group. Meanwhile, another 10 samples of lumbar nucleus pulposus served as controls. Hematoxylin-eosin staining and streptavidin-biotin peroxidase complex immunohistochemical technique were used to observe the expression of hypoxia inducible factor-1α in the human lumbar nucleus pulposus in different groups. RESULTS AND CONCLUSION:The expression level of hypoxia inducible factor-1α was (58.2±7.5)% in the sequestered group, (27.3±2.3)% in the protruded group, and (10.5±4.7)% in the control group, which was significantly higher in the sequestered group than the other two groups (P < 0.01). These findings indicate that the expression of hypoxia inducible factor-1α in the lumbarnucleus pulposus is associated with the herniated types, which is the highest in the prolapse sequestered type.

Objective:To observe the clinical effect of acupuncture combined with low-frequency electric stimulation on scissor gait in children with spastic cerebral palsy. Methods:A total of 60 spastic cerebral palsy kids were allocated into two groups by random number table, 30 in each group. Cases in the control group were treated with physical exercise therapy, massage and hydrotherapy. Based on the therapies given to the control group, cases in the observation group were supplemented with acupuncture combined with low-frequency electric stimulation. Selected points included Zusanli (ST 36), Sanyinjiao (SP 6), Yanglingquan (GB 34), Jiexi (ST 41), Chengshan (BL 57), Naoqing [Extra, locates at 2 cun directly above Jiexi (ST 41)] and Genping (Extra, locates at the midpoint of the line connecting medial and lateral malleolus). The treatment was done once every other day and 10 d made up a treatment course. The low-frequency electric stimulation was applied to anterior tibia, 6 times a week and 20 times made up a treatment course. Cases were treated for a total of 3 courses and there was a 15-20 d interval between two courses. The low limb functions were assessed before and after treatment using the modified Ashworth scale (MAS) and composite spasticity scale (CSS). In addition, the ankle dorsiflexion angles were measured before and after treatment. Results:After treatment, the ankle dorsiflexion angles and CSS scores in the observation group were better than those in the control group, showing statistical significances (P<0.05); the effective rate in the observation group calculated by MAS and ankle dorsiflexion angle were both higher than that in the control group, showing a statistical significance (P<0.05). Conclusion:Rehabilitation training combined with acupuncture and low-frequency electric stimulation can achieve better effect than rehabilitation training alone in improving scissor gait in kids with spastic cerebral palsy.