1.Using Image-Pro Plus combined with Photoshop image analysis software for quantitative measurement of changes in hippocampus microcirculation
Luping HUANG ; Yunchang MO ; Qinxue DAI ; Shenhui JIN ; Junlu WANG
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care 2015;(6):587-589,590
Objective To investigate a new method using Image-Pro Plus (IPP) combined with Photoshop image analysis software to quantitatively measure the changes in microcirculation in hippocampus.Methods Twenty-two Japanese white rabbits that had received bilateral carotid artery ligation for 2 weeks without neurological dysfunction were divided into a subarachnoid hemorrhage (SAH) group and a control group, 11 rabbits in each group. The rabbit model of symptomatic cerebral vasospasm was established by the method of twice injecting blood into occipital cistern. On the 7th day after the first time of injecting blood, the rabbits were sacrificed, cerebral perfusion fixation was carried out, the hippocampus was harvested, and CD34 was determined by immunohistochemical determination. IPP 6.0 combined with Photoshop image analysis software was used to quantitatively measure the count of hippocampal microvessels density (MVD) and the field (for statistics)/microvascular capillary area ratio was calculated.Results CD34 could effectively identify microvascular endothelial cells, and using IPP 6.0 could automatically and accurately calculate MVD and field (for statistics)/micrevascular area ratio in hippocampus. Compared with the control group, in SAH group, the MVD and the area ratio in hippocampus were significantly reduced, the differences being statistically significant [MVD (count/area): 3.87±0.67 vs. 5.17±0.53, area ratio: (0.86±0.20)% vs. (1.40±0.17)%, bothP < 0.05].Conclusions CD34 can be used to identify microvessels, IPP 6.0 image analysis software co-Photoshop is a high efficient and accurate new method to measure the microvessels count and calculate the field/microvessel area ratio, not only it is easy to operate, but also the data can be automatically calculated and generated, reflecting precisely the changes in microcirculation in hippocampus after symptomatic cerebral vasospasm in rabbits.
2.Influence of ginsenoside Rb1 pretreatment on expression of brain derived neurotrophic factor in rat hippocampus after acute immobilization stress
Qinxue DAI ; Junlu WANG ; Yuanyuan PAN ; Shenhui JIN ; Yunchang MO ; Danyun JIA
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care 2017;24(1):82-84
Objective To investigate the effects of ginsenoside Rb1 pretreatment on the expression of brain derived neurotrophic factor (BDNF) in hippocampus of rat models under acute immobilization stress.Methods Eighteen Sprague-Dawley (SD) rats were randomly divided into three groups (each n =6):normal control group,acute immobilization stress model group,and ginsenoside Rbl group.The rats in acute immobilization stress model group and ginsenoside Rb1 group were exposed to acute immobilization for 2 hours.Thirty minutes before the modeling,ginsnoside Rb1 (40 mg/kg) was injected intraperitoneally into rats in the ginsenoside Rbl group,and the control group was not treated.The enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of plasma cortisol (CORT) and adrenocorticotropic hormone (ACTH).The real-time fluorescence quantitative reverse transcription-polymerase chain reaction (RT-PCR) was applied to examine the expression of BDNF mRNA in rat hippocampus and its expression of BDNF protein was measured by Western Blot.Results In acute immobilization stress model group,compared with those before modeling,the plasma CORT and ACTH concentrations were significantly higher after modeling [CORT (μg/L):3.79 ± 0.50 vs.2.06 ± 0.35,ACTH (μg/L):1.69 ± 0.12 vs.0.94 ± 0.12,both P <0.05];compared with the normal control group,the mRNA and protein expressions of BDNF in hippocampus in the acute immobilization stress model group were decreased significantly [BDNF mRNA (A value):42.87 ± 5.56 vs.109.39 ± 9.11,BDNF protein (grey value):0.94 ± 0.02 vs.1.02 ± 0.03,both P < 0.01];compared with acute immobilization stress model group,the mRNA (113.73 ± 6.24 vs.42.87 ± 5.56) and protein expressions (1.04 ± 0.02 vs.0.94 ± 0.02) of BDNF in hippocampus of pre-treatment groups were significantly higher (all P < 0.05).Conclusions The results suggest that pretreatment with ginsenoside Rb1 alleviate hippocampus lesion induced by acute immobilization stress through regulating the BDNF mRNA and protein expressions in hippocampus.
3.The effect of Shenmai injection on purine content in rat's cerebral tissue
Luping HUANG ; Xin HE ; Qinxue DAI ; Shuangdong CHEN ; Sijia CHEN ; Junlu WANG
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care 2015;26(2):154-156
Objective To investigate the effect of Shenmai injection on purine content in rat cerebral cortex in order to provide a theoretical basis concerning its brain protective mechanism. Methods Sixteen Sprague-Dawley (SD) rats were randomly divided into two groups:normal saline control group and Shenmai injection group, with 8 rats in each group. Shenmai injection 15 mL/kg was injected intraperitoneally into the rats in Shenmai injection group, while in the normal saline group, an equal volume of normal saline was intraperitoneally injected. After the injection for 24 hours, the rats were sacrificed, and the cerebral cortex was removed on ice, homogenized and its supernatant was extracted;then high performance liquid chromatography (HPLC) was used to detect adenosine triphosphate (ATP), adenosine diphosphate (ADP), adenosine monophosphate (AMP), adenosine and inosine contents in the supernatant of cerebral cortex. Results Compared with normal saline control group, ATP, ADP, AMP, adenosine and creatinine content in the cerebral cortex of Shenmai injection group were significantly higher, the differences being statistically significant [ATP (ng/L): 31.62±5.12 vs. 20.25±4.53, ADP (ng/L): 37.04±6.72 vs. 25.12±7.35, AMP (ng/L): 87.82±20.37 vs. 33.23±10.34, adenosine (ng/L): 2.82±0.15 vs. 1.12±0.61, creatinine (ng/L): 11.72±1.05 vs. 6.05±2.55, P < 0.05 or P<0.01]. Conclusion Shenmai injection can elevate ATP, ADP, AMP, adenosine and creatinine contents in the cerebral cortex of rats, possibly that is the theoretical basis for brain protective mechanism of Shenmai injection.
4.Effect and mechanism of dexmedetomidine combined with sufentanil on postoperative analgesia in elderly patients with abdominal surgery
Mingxiao ZHANG ; Shenhui JIN ; Shuangdong CHEN ; Qinxue DAI ; Luping HUANG ; Sijia CHEN ; Junlu WANG
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care 2018;25(1):44-48
Objective To study the effect and mechanism of dexmedetomidine combined with sufentanil on postoperative analgesia in elderly patients with abdominal surgery. Methods Ninety-six elderly patients having undergone abdominal surgery in the First Affiliated Hospital of Wenzhou Medical University from January 2016 to June 2017 were enrolled, and they were divided into a control group and an observation group by random number table method, 48 cases in each group. General anesthesia was performed in the operation, and after surgery venous analgesic pump was used for analgesia in both groups. Analgesic method: the control group was given sufentanil 2 μg/kg and tropisetron 5 mg; the observation group was given dexmedetomidine 2 μg/kg, sufentanil 2 μg/kg and tropisetron 5 mg. The changes of pain and sedation score, conventional indexes [systolic blood pressure (SBP), diastolic blood pressure (DBP), heart rate (HR), respiratory rate (RR), pulse blood oxygen saturation (SpO2)], oxidative damage indexes [lipid peroxide (LPO), glutathione peroxidase (GSH-Px), Cu-Zn superoxide dismutase (Cu-Zn SOD)], stress response indexes [cortisol (Cor), epinephrine, norepinephrine (NE)], platelet activation indexes granule membrane protein-140 (GMP-140), thromboxane A2(TXA2) and the incidence of adverse reactions were observed in both groups. Results ① After surgery the visual simulation score (VAS) and Ramsay score in both groups were higher than those before surgery, and showed a tendency firstly increased and then decreased, and reached to peak value 2 hours after operation [VAS score:the control group was 3.24±0.98 vs. 1.95±0.93, observation group was 3.19±1.03 vs. 1.98±0.95; Ramsay score:the control group was 3.26±0.51 vs. 1.90±0.45, observation group was 3.77±0.53 vs. 1.92±0.42], began to decline 6 hours after operation, reached to valley value 48 hours after operation, and there was no significant difference in VAS scores between the two groups (2.02±0.64 vs. 1.98±0.95), Ramsay score was significantly higher in observation group than that in control group (2.59±0.41 vs. 2.10±0.21). ② Since 2 hours after the operation, the SBP, DBP, HR and RR in the observation group began to be lower than those in control group, 12 hours after surgery their values reached their valleys [SBP (mmHg, 1 mm Hg = 0.133 kPa): 113.2±13.5 vs. 122.1±10.3, DBP (mmHg): 67.5±9.9 vs. 76.4±8.6, HR (bpm): 64.5±6.9 vs. 71.4±7.5, RR (bpm): 14.8±1.1 vs. 15.8±0.8, all P < 0.05]; while SpO2did not change a great deal. ③ LPO, Cor, epinephrine, NE, GMP-140, TXA2of two groups after operation were higher than those before operation, GSH-Px and Cu-Zn SOD were lower than those before operation. However, LPO, Cor, epinephrine, NE, GMP-140, TXA2in observation group were significantly lower than those in control group [LPO (μmol/L): 6.4±0.8 vs. 9.8±1.1, Cor (ng/L): 148.2±19.1 vs. 239.3±27.8, epinephrine (μg/L): 124.2±13.9 vs. 207.1±23.5, NE (μg/L): 109.2±14.8 vs. 183.3±21.6, GMP-140 (μg/L): 27.13±3.82 vs. 39.06±4.83, TXA2(ng/L): 422.30±53.74 vs. 610.43±73.21, all P < 0.05], GSH-Px and Cu-Zn SOD in observation group were significantly higher than those in the control group [GSH-Px (U/L): 426.7±58.7 vs. 307.9±51.2, Cu-ZnSOD (μg/L): 311.3±42.5 vs. 231.6±34.1, all P < 0.05].④ The incidence of adverse reaction nausea in the observation group was significantly lower than that in the control group [4.17% (2/48) vs. 37.5% (18/48)]. Conclusion Dexmedetomidine combined with sufentanil used in elderly patients after abdominal surgery has significant analgesic effect, can effectively inhibit platelet activation, and decrease the contents of GMP-140 and TXA2.