Effect of electroacupuncture on calcium homeostasis in hippocampal neurons of mice with sepsis-associated encephalopathy / 中华麻醉学杂志

Objective:To investigate the effect of electroacupuncture on calcium homeostasis in hippocampal neurons of mice with sepsis-associated encephalopathy (SAE).Methods:Twenty-four healthy male C57BL/6J mice, weighing 18-22 g, were divided into 4 groups ( n=6 each) using a random number table method: sham operation group (Sham group), SAE group, SAE plus electroacupuncture group (SAE+ EA group), and SAE plus sham electroacupuncture group (SAE+ SEA group). The virus carrying calcium ion (Ca 2+ ) fluorescent probes was injected and then an optical fiber was implanted into the hippocampal CA1 area to record the fluorescence signals of Ca 2+ . SAE was induced by cecal ligation and puncture in anesthetized mice at 3 weeks after administration. Starting from 3 days before surgery, Baihui and bilateral Quchi and bilateral Zusanli acupoints were stimulated for 30 min per day for 7 consecutive days in SAE+ EA group. In SAE+ SEA group, electroacupuncture was performed at the points 0.2 mm lateral to the corresponding acupoints without electrical stimulation. Open field tests were conducted at 5 days after surgery to record the number of rearing and changes in related Ca 2+ signals in hippocampal CA1 neurons. Novel object recognition tests were conducted at 6-7 days after surgery to record the recognition time and changes in related Ca 2+ signals in hippocampal CA1 neurons. Mice were sacrificed after the end of behavioral testing on 7 days after surgery, and brain tissues ipsilateral to the optical fiber implant were obtained and the fluorescence intensity of Ca 2+ in the hippocampal CA1 neurons was acquired using a fluorescent microscope. Results:Compared with Sham group, the number of rearing and amplitudes of related Ca 2+ signals in hippocampal CA1 neurons while rearing were significantly decreased in SAE group and SAE+ SEA group ( P<0.05), and no statistically significant changes were found in the parameters mentioned above in SAE+ EA group ( P>0.05), and the recognition index and amplitudes of related Ca 2+ signals while recognizing were significantly deceased, and the fluorescence intensity of Ca 2+ in hippocampal CA1 neurons was increased in SAE, SAE+ EA and SAE+ SEA groups ( P<0.05). Compared with SAE group and SAE+ SEA group, the number of rearing and amplitudes of related Ca 2+ signals in hippocampal CA1 neurons while rearing were significantly increased, the recognition index and amplitudes of related Ca 2+ signals in hippocampal CA1 neurons while recognizing were increased, and the fluorescence intensity of Ca 2+ in hippocampal CA1 neurons was decreased in SAE+ EA group ( P<0.05). There were no statistically significant differences in the parameters mentioned above between SAE group and SAE+ SEA group ( P>0.05). Conclusions:The mechanism by which electroacupuncture alleviates SAE may be related to regulation of Ca 2+ homeostasis in hippocampal neurons of mice.

Role of SIRT1 in mitochondrial dysfunction in mice with lipopolysaccharide-induced brain injury / 中华麻醉学杂志

Objective:To evaluate the role of silent information regulator sirtuin 1 (SIRT1) in mitochondrial dysfunction in mice with lipopolysaccharide (LPS)-induced brain injury.Methods:Eighty clean-grade male C57BL/6 mice, aged 6-8 weeks, were divided into 4 groups ( n=20 each) by the random number table method: control group (group C), LPS-induced brain injury group (LPS group), LPS-induced brain injury plus SIRT1 inhibitor EX527 group (LPS+ E group), and LPS-induced brain injury plus SIRT1 agonist SRT1720 group (LPS+ S group). Brain injury was induced by intravenous injection of LPS 10 mg/kg. EX527 10 mg/kg was intraperitoneally injected at 72 h before LPS injection in group LPS+ E, and the equal volume of dimethyl sulfoxide was intraperitoneally injected instead in the other three groups. SRT1720 100 mg/kg was intraperitoneally injected at 30 min before LPS injection in group LPS+ S, and the equal volume of dimethyl sulfoxide was intraperitoneally injected instead in the other three groups. The novel object recognition test was performed at 24 h after LPS injection, then the mice were sacrificed, and hippocampal tissues were harvested for determination of the number of the normal neurons in the hippocampal CA1 area, ATP content and activities of mitochondrial respiratory chain complexes Ⅰ, Ⅱ, Ⅲ and Ⅳ (by spectrophotometry), and mitochondrial membrane potential (MMP) (by Jc-1 staining) and for microscopic examination of pathological changes (by Nissl staining) and ultrastructure of neuronal mitochondria (with a transmission electron microscope). Results:Compared with group C, the preference index in novel object recognition, normal neuron count, activities of mitochondrial respiratory chain complexes Ⅰ, Ⅱ, Ⅲ and Ⅳ, MMP and ATP content were significantly decreased ( P<0.05), damage to hippocampal neurons was found, mitochondrial swelling was observed and cristae structure ruptured in LPS, LPS+ S and LPS+ E groups. Compared with group LPS, the preference index in novel object recognition, activities of mitochondrial respiratory chain complexes, MMP and ATP content were significantly decreased ( P<0.05), neuronal damage was aggravated, the mitochondrial swelling and fracture of crista structure were accentuated in group LPS+ E; the preference index in novel object recognition, activities of mitochondrial respiratory chain complexes, MMP and ATP content were significantly increased ( P<0.05), neuronal damage was alleviated, and the mitochondrial swelling and fracture of crista structure were ameliorated in group LPS+ S. Conclusions:Activation of SIRT1 can improve mitochondrial dysfunction and alleviate LPS-induced brain injury in mice.

A retrospective study of cisatracurium besilate-assisted treatment of severe acute pancreatitis complicated with intraabdominal hypertension / 中华麻醉学杂志

The clinical data of patients with severe acute pancreatitis complicated with intraabdominal hypertension or abdominal compartment syndrome admitted to our Department of Critical Care Medicine from January 1, 2018 to October 1, 2021 were collected and analyzed.Patients were divided into a conventional treatment group and conventional treatment plus IV infusion of cisatracurium besilate group (muscle relaxation group). A prediction model of treatment propensity score was developed for paired screening, with 31 cases in each group.The conventional treatment group adopted conventional basic treatment methods such as gastrointestinal decompression, spasmolysis and analgesia, fluid therapy, inhibition of gastric acid, suppression of parenzyme, nutritional support, mechanical ventilation, and enemata.In muscle relaxation group, cisatracurium besilate was intravenously infused on the basis of routine treatment with the initial dose of 0.15 mg/kg given to facilitate endotracheal intubation, followed by continuous intravenous infusion at 1-3 μg·kg -1·min -1, and the dose was adjusted according to the patient′s basic vital signs and clinical effects.The primary outcome was survival rate.Secondary outcome measures were changes in intraabdominal pressure, oxygenation index, the number of defecation, volume of defecation, and urination volume before treatment and on 7, 14 and 20 days of treatment.and the recovery time of bowel sounds, length of mechanical ventilation, time of intensive care unit treatment, and total hospitalization costs.Compared with conventional treatment group, the survival rate was significantly increased, the intraabdominal pressure was decreased on 7, 14 and 20 days of therapy, the oxygenation index was increased, the number of defecation and volume of defecation were increased on 7 and 14 days of therapy, urinary volume was increased before treatment and on day 7 of therapy, the recovery time of intestinal sound was significantly shortened ( P<0.05), and no significant change was found in urinary volume on days 14 and 20 of therapy, length of ventilation, time of intensive care unit treatment, and total hospitalization costs in muscle relaxation group ( P>0.05). In conclusion, cisatracurium besilate can improve oxygenation, promote the recovery of intestinal function and improve the survival rate when used to assist the treatment in the patients with severe acute pancreatitis complicated with intraabdominal hypertension.

Relationship between HO-1 and PPARγ during alveolar macrophage polarization in a mouse model of endotoxin-induced acute lung injury / 中华麻醉学杂志

Objective:To evaluate the relationship between heme oxygenase-1 (HO-1) and peroxisome proliferator-activated receptor γ (PPARγ) during alveolar macrophage polarization in a mouse model of endotoxin-induced acute lung injury (ALI).Methods:Thirty clean-grade male C57BL/6 mice (24 wide-type mice and 6 HO-1 knockout mice), aged 6-8 weeks, weighing 18-22 g, were studied.Wide-type mice were divided into 4 groups ( n=6 each) using a random number table method: control group (C group), ALI group, ALI+ HO-1 agonist hemin group (ALI+ H group), and ALI+ hemin+ PPARγ antagonist T0070907 group (ALI+ H+ T group).HO-1 knockout mice in which the ALI model was developed served as ALI+ HO-1 -/- group.ALI model was developed by injecting lipopolysaccharide (LPS) 15 mg/kg via the tail vein in anesthetized animals.T0070907 1.5 mg/kg was intraperitoneally injected at 1 h before LPS administration in ALI+ H+ T group, and hemin 50 mg/kg was intraperitoneally injected at 30 min before LPS administration in ALI+ H group and ALI+ H+ T group.Mice were sacrificed at 12 h after LPS administration, and lung tissues were obtained to measure the wet to dry weight ratio (W/D ratio), to observe pathological changes which were scored, and to determine the F4/80+ /CD86+ labeled M1 alveolar macrophages and the F4/80+ /CD206+ labeled M2 alveolar macrophages (by flow cytometry), contents of M1 macrophage-related genes inducible nitric oxide synthase (iNOS) and M2 macrophage-related genes Arginase-1 (Arg-1) (by enzyme-linked immunosorbent assay), and the expression of HO-1 and PPARγ (by Western blot). Results:Compared with C group, the lung injury score, W/D ratio, levels of CD86 and CD206, and contents of iNOS and Arg-1 were significantly increased, and PPARγ expression was up-regulated in the other four groups ( P<0.05), and HO-1 protein expression was up-regulated in ALI, ALI+ H and ALI+ H+ T groups ( P<0.05).Compared with ALI group, the lung injury score, W/D ratio, and levels of CD86 and iNOS were significantly increased, the levels of CD206 and Arg-1 were decreased, and the expression of HO-1 and PPARγ was down-regulated in ALI+ HO-1 -/- group, the lung injury score, W/D ratio and levels of CD86 and iNOS were significantly decreased, the levels of CD206 and Arg-1 were increased, and the expression of HO-1 and PPARγ was up-regulated in ALI+ H group ( P<0.05), and no significant change was found in the parameters mentioned above in ALI+ H+ T group ( P>0.05).Compared with ALI+ H group, the lung injury score, W/D ratio and levels of CD86 and iNOS were significantly increased, the levels of CD206 and Arg-1 were decreased, the expression of PPARγ was down-regulated ( P<0.05), and no significant change was found in the expression of HO-1 in ALI+ H+ T group ( P>0.05). Conclusions:HO-1 can up-regulate the expression of PPARγ, inhibit the polarization of alveolar macrophages toward M1 phenotype and promote the polarization toward M2 phenotype, thus playing an endogenous protective role in endotoxin-induced ALI in mice.

Effect of electroacupuncture pretreatment on oxidative stress response of hippocampus in rats with sepsis-associated encephalopathy and relationship with Nrf2/HO-1 signaling pathway / 中华麻醉学杂志

Objective:To evaluate the effect of electroacupuncture (EA) pretreatment on oxidative stress response of hippocampus in rats with sepsis-associated encephalopathy (SAE) and the relationship with nuclear factor E2-related factor 2 (Nrf2)/heme oxygenase-1 (Nrf2/HO-1) signaling pathway.Methods:A total of 64 healthy adult male Sprague-Dawley rats, weighing 250-300 g, were divided into 4 groups ( n=16 each) using a random number table method: sham operation group (Sham group), SAE group, SAE+ EA group and SAE plus sham EA group (SAE+ SEA group). In SAE+ EA group, Baihui, Quchi and Zusanli acupoints were stimulated for 30 min once a day for 5 consecutive days.Sepsis was induced by cecal ligation and puncture immediately after the end of the last EA.At 1 and 7 days after establishment of the model, the hippocampal malondialdehyde (MDA) and superoxide dismutase (SOD) levels were detected by enzyme-linked immunosorbent assay, the expression of hippocampal Nrf2 mRNA was detected using real-time reverse transcription polymerase chain reaction, and the expression of Nrf2 and HO-1 was determined by Western blot.At 3-7 days after establishment of the model, cognitive function was assessed using the Morris water maze test, and the escape latency and the target quadrant exploration time were recorded. Results:Compared with Sham group, the content of MDA was significantly increased and the activity of SOD was decreased at 1 and 7 days after establishment of the model, the expression of Nrf2 protein and mRNA and HO-1 was down-regulated at day 7 after establishment of the model, the escape latency was prolonged, and the target quadrant exploration time was shortened in SAE group ( P<0.05). Compared with SAE group, the content of MDA was significantly decreased and the activity of SOD was increased at 1 and 7 days after establishment of the model, the expression of Nrf2 protein and mRNA and HO-1 was up-regulated at day 7 after establishment of the model, the escape latency was shortened, and the target quadrant exploration time was prolonged in group SAE+ EA ( P<0.05), and no significant change was found in the parameters mentioned above in group SAE+ SEA ( P>0.05). Conclusion:EA pretreatment can reduce oxidative stress response of hippocampus in rats with SAE, and the mechanism may be related to activating Nrf2/HO-1 signaling pathway.

Effect of electroacupuncture on synaptic damage to hippocampal neurons in rats with sepsis-associated encephalopathy / 中华麻醉学杂志

Objective:To investigate the effect of electroacupuncture (EA) on synaptic damage to hippocampal neurons in rats with sepsis-associated encephalopathy (SAE).Methods:A total of 48 healthy adult male Sprague-Dawley rats, weighing 250-300 g, were divided into 4 groups ( n=12 each) using a random number table method: sham operation group (Sham group), SAE group, SAE+ EA group and SAE+ sham EA group (SAE+ SEA group). SAE was induced by cecal ligation and puncture in anesthetized rats.Baihui, Quchi and Zusanli acupoints were stimulated with constant voltage (2/15 Hz) and disperse-dense waves for 30 min once a day for 10 consecutive days, and the stimulation intensity was defined as less than 1.5 mA causing slight muscle contraction at 2 days before operation in group SAE+ EA.In group SAE+ SEA, stimulating electrodes were placed at the points 5 mm lateral to the corresponding acupoints, but no electrical stimulation was applied.On day 14 after operation, the rats were sacrificed, and hippocampal tissues were obtained and stained with haematoxylin and eosin for examination of the pathological changes in the hippocampal CA1 region, for determination of the expression of synaptophysin (SYN) and postsynaptic density protein 95 (PSD-95) (by Western blot), and for calculation of dendritic spine density of neurons in the hippocampal CA1 area (using Golgi staining) and pyramidal neurons counts. Results:Compared with Sham group, the expression of SYN and PSD-95 in hippocampus was significantly decreased, and the basal and apical dendrite spine density of neurons in hippocampal CA1 area was decreased in SAE group, the expression of PSD-95 was decreased, and the apical dendrite spine density of neurons in the hippocampal CA1 area was increased in SAE+ EA group, and the pyramidal neuron counts in the hippocampal CA1 area were reduced in SAE, SAE+ EA and SAE+ SEA groups ( P<0.05). Compared with group SAE, the expression of SYN and PSD-95 was significantly up-regulated, the basal and apical dendrite spine density of neurons in the hippocampal CA1 area was increased and the pyramidal neuron counts were increased in group SAE+ EA ( P<0.05), the pathological damage of hippocampal CA1 area was alleviated and no significant change was found in the parameters mentioned above in group SAE+ SEA ( P>0.05). Compared with group SAE+ EA, the expression of SYN and PSD-95 was down-regulated, the basal and apical dendrite spine density of neurons in the hippocampal CA1 area was decreased, and the pyramidal neuron counts were reduced in SAE+ SEA group ( P<0.05). Conclusion:The mechanism by which EA alleviates SAE may be related to reducing synaptic damage to hippocampal neurons in rats.

Role of HO-1 in electroacupuncture-induced alleviation of cognitive dysfunction in mice with sepsis-associated encephalopathy: relationship with mitochondrial fusion-division balance / 中华麻醉学杂志

Objective:To evaluate the role of heme oxygenase-1 (HO-1) in electroacupuncture (EA)-induced alleviation of cognitive dysfunction in mice with sepsis-associated encephalopathy (SAE) and the relationship with mitochondrial fusion-division balance.Methods:Thirty clean-grade male C57BL/6 mice (24 wide-type mice and 6 HO-1 knockout mice), aged 6-8 weeks, weighing 18-22 g, were studied.Twenty-four wide-type mice were divided into 4 groups ( n=6 each) using a random number table method: control group (group C), SAE group, SAE plus EA group (group SAE+ EA), and SAE plus sham EA group (group SAE+ SEA). HO-1 knockout mice in which EA intervention was performed after establishing SAE model served as SAE plus EA plus HO-1 knockout group (group SAE + EA+ H). Sepsis was induced by intraperitoneally injecting lipopolysaccharide 15 mg/kg.EA of Zusanli and Baihui acupoints lasting 30 min was performed after intraperitoneal injection of lipopolysaccharide once a day for 5 consecutive days in SAE+ EA and SAE+ EA+ H groups.Cognitive function was assessed using Morris water maze test before stimulation every day.The mice were sacrificed, and hippocampal tissues were removed for detection of the expression of mitofusin 2 (Mfn2), optic atrophy 1 (OPA1) and mitochondrial dynamin-related protein 1 (Drp1) by Western blot. Results:Compared with group C, the expression of Mfn2 and OPA1 was significantly down-regulated, the escape latency was prolonged, and the time spent in the target quadrant was shorted in SAE, SAE+ SEA and SAE+ EA+ H groups, and the expression of Drp1 was significantly up-regulated in SAE, SAE+ EA, SAE+ SEA and SAE+ EA+ H groups ( P<0.05). Compared with group SAE, the expression of Mfn2 and OPA1 was significantly up-regulated, the expression of Drp1 was down-regulated, the escape latency was shortened, and the time spent in the target quadrant was prolonged in group SAE+ EA ( P<0.05), and no significant change was found in the parameters mentioned above in group SAE+ SEA ( P>0.05). Compared with group SAE+ EA, the expression of Mfn2 and OPA1 was significantly down-regulated, the expression of Drp1 was up-regulated, the escape latency was prolonged, and the time spent in the target quadrant was shortened in SAE+ SEA and SAE+ EA+ H groups ( P<0.05). Conclusion:HO-1 is involved in EA-induced alleviation of cognitive dysfunction in mice with SAE, and the mechanism may be related to the regulation of mitochondrial mitochondrial fusion-division balance.

The diversity of endophytic fungi in endangered medicinal material Daphnes Cortex as determined by high-throughput sequencing technology / 药学学报

In order to provide a scientific basis for the establishment of a Daphnes Cortex medicinal material fungus library and the screening of endophytic fungi that promote the growth of Daphnes Cortex and increase the content of daphnetin, we used Illumina high-throughput testing technology to analyze 9 Daphnes Cortex samples from Gansu and Shanxi provinces. A total of 632 766 valid sequences were obtained, including 348 OTUs, 4 phyla, 20 classes, 48 orders, 108 families, 154 genera, and 208 species. The sum of the first 3 fungal genera account for more than 65% of the total abundance, with the highest reaching 98.4%. Alternaria and Phoma are the main genuses of Daphne giraldii Nitsche, and Altemaria is the dominant genus. The endophytic fungi community of Daphnes Cortex is rich in diversity, and the order of fungal diversity in different producing areas is: Gangu County > Wutai County > Tanchang County.

Role of hippocampal HO-1 in electroacupuncture-induced reduction of lipopolysaccharide-induced brain injury in mice / 中华麻醉学杂志

Objective:To evaluate the role of hippocampal hemeoxygenase-1 (HO-1) in electroacupuncture (EA)-induced reduction of lipopolysaccharide (LPS)-induced brain injury in mice.Methods:Twenty-four healthy adult C57BL/6J mice of both sexes, weighing 18-22 g, were divided into 4 groups ( n=6 each) using a random number table method: control group (group C), LPS-induced brain injury group (LPS group), LPS plus EA group, and LPS plus EA plus HO-1 inhibitor zinc protoporphyria (ZnPP) group (LPS+ EA+ ZnPP group). A virus carrying calcium ion fluorescent probes was injected into and an optical fiber was implanted into the hippocampal CA1 region to record changes in the calcium fluorescence signals.Three weeks later, Baihui, Quchi and Zusanli acupoints were stimulated with constant voltage (2/15 Hz) and disperse-dense waves for 30 min once a day for 5 consecutive days, and the stimulation intensity was defined as less than 1 mA causing slight muscle contraction.ZnPP 50 μmol/kg was intraperitoneally injected at 12 h before each stimulation in LPS+ EA+ ZnPP group, and the equal volume of normal saline was given instead in the other groups.After the end of EA stimulation on the last day, LPS 5 mg/kg was intraperitoneally injected to induce brain injury.Open field tests were performed at 1 day after LPS injection to record the number of rearing and amplitude of neuronal calcium signals during rearing.Novel object recognition tests were conducted at 3 days after LPS injection, and the exploration index and amplitude of neuronal calcium signals while exploring novel objects were recorded.The mice were sacrificed after the end of behavioral testing, and the brain tissues were obtained and stained by Nissl, and the neurons in the hippocampal CA1 region were counted. Results:Compared with group C, the number of rearing and amplitude of calcium signals in neurons in the hippocampal CA1 region during rearing were significantly decreased, the exploration index and amplitude of calcium signals in neurons in the hippocampal CA1 region while exploring novel objects were decreased, and the neuron counts were reduced in LPS, LPS+ EA and LPS+ EA+ ZnPP groups ( P<0.05 or 0.01). Compared with group LPS, the number of rearing and amplitude of calcium signals in neurons in the hippocampal CA1 region during rearing were significantly increased, and the exploration index and amplitude of calcium signals in neurons in the hippocampal CA1 region while exploring novel objects were increased in group LPS+ EA ( P<0.05), and no significant change was found in the parameters mentioned above in group LPS+ EA+ ZnPP ( P>0.05). Compared with group LPS+ EA, the number of rearing and amplitude of calcium signals in neurons in the hippocampal CA1 region during rearing were significantly decreased, and the exploration index and amplitude of calcium signals in neurons in the hippocampal CA1 region while exploring novel objects were decreased in group LPS+ EA+ ZnPP ( P<0.05). Conclusion:The mechanism by which EA reduces LPS-induced brain injury is related to the activation of the endogenous protective mechanism HO-1 in mice.

Preparation and cytocompatibility of porous zirconia scaffold modified by graphene oxide for bone tissue engineering / 医学研究生学报

Objective In order to meet the needs of maxillofacical bone defect repair, the aim of this study was to synthesize graphene oxide(GO) modified three-dimensional conneted nano- zirconia(ZrO2) bone tissue engineering scaffold and evaluate its surface morphology, compressive strength and cytocompatibility. Methods GO was synthesized by a modified Hummers method and then was testified by scanning electron microscope, transmission electron microscopy and fourier transform infrared spectroscopy. ZrO2 scaffold was modified by different concentrations(0.5,1.0,1.5mg/mL) of GO dispersion via a silane-mediated method. The composite scaffold with uniform GO coating was chosen for compressive strength test and co-cultured with human dental pulp stem cells(hDPSCs). Actin staining was used to observe the growth of the cells on the scaffold, and MTS was used to detect the cell activity. Results The characterization results showed that, under scanning electron microscope, the GO was flaky and the surface morphology of folds could be seen. Part of the GO layer folds up at the edge. Under transmission electron microscopy, the GO was clearly observed to have a gossylike, translucent and slightly wrinkled lamellar structure. The crystal structure in this area in the high-resolution filter image showed a six-member ring structure like graphite. Under high power electron microscope, the 1.0mg/ml GO-ZrO2 scaffold could be seen to deposit a thin layer of GO at the crack of the scaffold skeleton, connecting the two ends of the crack, and lamellar GO with folds could be observed on the surface of ceramic particles. The comparison of mechanical properties showed that the compression strength of GO-ZrO2 scaffold was sgnificantly increased compared with that of ZrO2 scaffold[（1.292±0.087）vs（1.031±0.076）, P<0.05]. Compared with the simple ZrO2 scaffold, the GO-ZrO2 scaffold showed more dense extension and adhesion to the surface of scaffolds, showing more active cell proliferation. The cell viability test showed that the viability of hDPSCs was significantly improved on GO-ZrO2 scaffold after 1, 3 and 5 days of proliferation compared with the simple ZrO2 scaffold(P<0.05). Conclusion The ZrO2 scaffold modified by GO improved compressive strength, promoted the early proliferation of hDPSCs with good cytocompatibility.

Role of endogenous heme oxygenase-1/carbon monoxide signaling pathway in endoplasmic reticulum stress during endotoxin-induced acute lung injury in rats / 中华麻醉学杂志

Objective To evaluate the role of endogenous heme oxygenase-1/carbon monoxide ( HO-1/CO) signaling pathway in endoplasmic reticulum stress during endotoxin-induced acute lung injury ( ALI) in rats. Methods Forty healthy clean-grade male Sprague-Dawley rats, aged 8 weeks, weighing 190-210 g, were divided into 4 groups ( n=10 each) using a random number table method: control group (group C), ALI group, ALI plus ZnPP-IX group (group AZ), and ALI plus vehicle sodium bicarbonate group ( group AV) . ALI was induced by intravenously injecting lipopolysaccharide 5 mg/kg in anesthetized rats. At 30 min before establishing the model, ZnPP-IX 10μmol/kg (diluted to 1 ml in 50 mmol/L sodium bicarbonate) was intraperitoneally injected in group AZ, and 50 mmol/L sodium bicarbonate 1 ml was intra-peritoneally injected in group AV. After injecting lipopolysaccharide for 6 h, blood samples were collected from the common carotid artery for determination of plasma CO concentration, the rats were then sacrificed, and lungs were removed for microscopic examination of the pathological changes which were scored and for determination of CO level, wet to dry weight ratio ( W/D ratio) , cell apoptosis ( by TUNEL) , and expres-sion of heme oxygenase-1 ( HO-1) , glucose-regulated protein 78 ( GRP78) , phosphorylated protein kinase R-like endoplasmie reticulum kinase (p-PERK), phosphorylated eukaryotic translation initiation factor 2 alpha ( p-elF2 ) , CCAAT/enhancer-binding protein homologous protein ( CHOP ) and caspase-12 in lung tissues ( by Western blot) . Apoptosis index ( AI) was calculated. Results Compared with group C, the lung injury scores, W/D ratio, AI and CO levels in plasma and lung tissues were significantly increased, and the expression of HO-1, GRP78, p-PERK, p-elF2, CHOP and caspase-12 was up-regulated in the other three groups ( P＜0. 05) . Compared with group ALI, lung injury scores, W/D ratio and AI were sig-nificantly increased, CO levels in plasma and lung tissues were decreased, the expression of HO-1 was down-regulated, and the expression of GRP78, p-PERK, p-elF2, CHOP and caspase-12 was up-regula-ted in group AZ (P＜0. 05), and no significant change was found in the parameters mentioned above in group AV ( P＞0. 05) . Conclusion HO-1/CO signaling pathway produces endogenous protection possibly through inhibiting endoplasmic reticulum stress during endotoxin-induced ALI in rats.

Study on the application of personalization customized oral positioning stent in head and neck cancer patients with intensity modulated radiation therapy / 医学研究生学报

Objective Head and neck cancer radiotherapy patients often appear a series of oral complications including mucositis, xerostomia, pain, dysphagia.The purpose of this study was to investigate whether personalization customized positioning oral stent was able to push normal tissue off the high dose target area and maintain accurate repeatable stable positions, thus protecting the normal tissue during radiotherapy of the nasopharyngeal carcinoma patients.Methods 15 newly diagnosed nasopharyngeal carcinoma patients were collected from March to August 2016 in Department of Radiation Oncology, Nanjing General Hospital of Nanjing Military Region and randomly divided into trial group and control group.Two groups of patients were treated with intensity modulated radiation therapy (IMRT).Trial group patients wear personalization customized oral positioning stents during radiotherapy while the control group did not wear.After radiotherapy, we compared the exposure doses of clinical target area(CTV) and normal oral tissue in two groups.ResultsThe left parotid gland radiotherapy doses of the trail group and the control group were 2223.557±294.549 cGy and 2900.563±374.660 cGy, the difference was statistically significant(t=3.847, P=0.002);the right parotid gland radiotherapy doses of the trail group and control group were 2284.957±256.673 cGy and 2994.670±339.264 cGy, the difference was statistically significant(t=4.512, P=0.001).The mean exposure doses of CTV in two groups were no statistically significant difference (6142.829±135.986 cGy vs 173.306±6221.825 cGy, t=0.971, P=0.349.Conclusion During the intensity modulated radiation therapy, patients with personalization customized oral positioning stents can keep the mandible in a precise repeatable stable position.And it can reduce the exposure dose of bilateral parotid without affect the radiotherapy effect of the clinical target area.

Influencing factors for duration of viral nucleic acid shedding in children with influenza A / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the features and duration of viral nucleic acid shedding in children with influenza A.</p><p><b>METHODS</b>The clinical data of 90 children with influenza A with positive influenza A virus nucleic acid in nasopharyngeal swab detected by PCR were collected, and these children were divided into simple influenza A group (n=10), influenza A-pneumonia group (n=61), influenza A-nervous system damage group (n=10), and influenza A-underlying disease group (n=9). A retrospective analysis was performed for clinical features, treatment process, duration of viral nucleic acid shedding, and prognosis.</p><p><b>RESULTS</b>The most common symptoms in these children were fever (89/90, 99%), cough (89/90, 99%), running nose (69/90, 77%), shortness of breath (26/90, 29%), and myalgia (23/90, 26%). The mean duration of viral nucleic acid shedding in 90 children was 9.4±2.9 days. The simple influenza A group had a significantly shorter duration of viral nucleic acid shedding than the influenza A-pneumonia, influenza A-nervous system damage, and influenza A-underlying disease groups (p<0.05), while there were no significant differences between the influenza A-pneumonia, influenza A-nervous system damage, and influenza A-underlying disease groups (p>0.05). The children who received antiviral therapy within 48 hours after disease onset had significantly shorter duration of viral nucleic acid shedding and time to body temperature recovery than those who received antiviral therapy more than 48 hours after disease onset (p<0.05). Of all the children with body temperature recovery, 83% still tested positive for viral nucleic acid.</p><p><b>CONCLUSIONS</b>Complications, underlying diseases, and timing of antiviral therapy are influencing factors for the duration of influenza A virus nucleic acid shedding, and whether body temperature returns to normal cannot be used to decide whether to continue antiviral therapy.</p>

Role of mitochondrial fusion-fission in endotoxin-induced acute lung injury in rats / 中华麻醉学杂志

Objective To evaluate the role of mitochondrial fusion?fission in endotoxin?induced a?cute lung injury in rats. Methods Twenty healthy male Sprague?Dawley rats, weighing 160-180 g, were e?qually and randomly divided into either control group ( group C ) or endotoxin?induced acute lung injury group (group L) using a random number table. Lipopolysaccharide 5 mg∕kg was injected intravenously in group L, while the equal volume of normal saline 0?5 ml was given instead in group C. The animals were sacrificed at 6 h after administration of lipopolysaccharide or normal saline. The lungs were immediately re?moved for measurement of wet to dry lung weight ratio ( W∕D ratio) , superoxide dismutase ( SOD) activity and malondialdehyde ( MDA) content. The mitochondrial fusion proteins mitofusin 1 ( Mfn1) , Mfn2 and op?tic atrophy 1 ( OPA1) mRNA and protein expression was detected, and mitochondrial fission proteins dy?namin?related protein 1 (Drp1) and fission 1 (Fis1) mRNA and protein expression was also detected in lung tissues. Results Compared to group C, the W∕D ratio and MDA contents in lung tissues were signifi?cantly increased, SOD activity was decreased, Mfn1, Mfn2 and OPA1 mRNA and protein expression in lung tissues was down?regulated, and Drp1 and Fis1 mRNA and protein expression was up?regulated in group L. The pathological damage to lung tissues was obviously aggravated in group L when compared to group C. Conclusion The mechanism underlying endotoxin?induced acute lung injury is related to enhanced oxidative stress responses caused by decreased mitochondrial fusion and increased mitochondrial fission in rats.

Role of spinal ERK signaling pathway in reduction of remifentanil-induced hyperalgesia by electro-acupuncture at Zusanli in rats with incisional pain / 中华麻醉学杂志

Objective To evaluate the role of spinal extracellular signal-regulated kinase (ERK) signaling pathway in reduction of remifentanil-induced hyperalgesia by electro-acupuncture (EA) at Zusanli in rats with incisional pain.Methods Fifty male adult Sprague-Dawley rats, weighing 250-280 g, in which the intrathecal catheter was successfully placed without complications, were randomly divided into 5 groups (n=10 each) using a random number table: control group (group C), remifentanil + incisional pain group (group RI), EA at acupoint group (group E) , EA at non-acupoint group (group NE), and 1/2 ERK inhibitor U0126 + EA at acupoint group (group UE).Normal saline 0.1 ml · kg-1 · min-1 was infused intravenously for 60 min in group C.In RI, E, NE and UE groups, after the model of incisional pain was established, remifentanil 1.0 μg · kg-1 · min-1 was infused for 60 min, and in addition, EA (intensity 10 mA, frequency 4 Hz) of Zusanli lasting for 60 min was performed at the same time in E and UE groups, and EA was performed at the points 5 mm lateral to the acupoints of Zusanli on the operated side simultaneously in group NE.ERK1/2 inhibitor U0126 5 μg (in 5％ dimethyl sulfoxide 10 μl) was injected intrathecally in group UE, and 5％ dimethyl sulfoxide 10 μl was injected intrathecally in the other groups.The mechanical paw withdrawal threshold (MWT) was measured before remifentanil or normal saline infusion (T1) , and at 2 h, 1, and 2 days after the end of infusion (T2-4).After MWT was measured at T4, the expression of ERK1/2 and phosphorylated ERK1/2 (p-ERK1/2) in spinal cord dorsal horns was measured by Western blot.Results Compared with group C, the MWT was significantly decreased at T2-4 in RI, E, NE and UE groups, and the expression of p-ERK1/2 was up-regulated in RI, E and NE groups (P＜0.05).Compared with group RI, the MWT was significantly increased at T2-4 , and the expression of p-ERK1/2 was down-regulated in E and UE groups (P＜0.05);and no significant change was found in the parameters mentioned above in group NE (P＞0.05).Compared with group E, the MWT was significantly increased at T2-4, and the expression of p-ERK 1/2 was down-regulated in group UE (P＜0.05).Conclusion The mechaism by which EA at Zusanli reduces hyperalgesia induced by remifentanil in rats with incisional pain is related to inhibited activation of ERK signaling pathway in the spinal cord.

Awaken Effect of Naloxon in Dexmedetomidine-Anesthetized Mice / 天津医药

Objective To investigate the awaken effect of naloxon on dexmedetomidine anesthetized mice and its mechanism. Methods Thirty Kunming mice of clean grade were randomly divided into 3 groups which included NAL group (Naloxon group), ATI group(Atipamezole group)and NS group (Normal Saline group). All groups were given dexme?detomidine 1 mg·kg-1 intraperitoneally. Naloxon 2 mg·kg-1, atipamezole 2 mg·kg-1 and normal saline 10 mL·kg-1 were ran?domly given intraperitoneally to the NAL, ATI and NS group respectively 90 minutes after dexmedetomidine administration. At timepoints prior to dexmedetomidine administration and 5, 15, 30, 60, 90, 95, 105, 120, 180 minutes after it, the sedative and analgesic effects besides recovery time (based on restore of righting reflex loss) were assessed. Results Sedation and analgesia effects became apparent within 5 minutes, and peaked at approximately 60 minutes then spontaneously recovered at 180 minutes after injection of dexmedetomidine. The sedative and analgesic effects were reduced in both ATI and NAL groups. Compared with ATI group, the sedation scores were higher at 95, 105 and 120 minutes after dexmedetomidine admin?istration than those in NAL group (P0.05). The analgesic scores were not statistically significant at time points of 95, 105, 120 and 180 min?utes between NAL group and ATI group, but they were lower in NAL group compared with NS group at timepoints of 95, 105 and 120 minutes (P>0.05). The recovery time in ATI and NAL group were shorter than that in NS group (F=1 793.368, P0.05). Conclusion Naloxone had a certain awaken effect on dexmedetomidine anesthetized mice.

Effect of electroacupuncture at Zusanli and Chize on sepsis-induced acute lung injury / 中华麻醉学杂志

Objective To evaluate the effects of electroacupuncture (EA) at Zusanli and Chize on sepsisinduced acute lung injury (ALI) in patients.Methods Sixty patients with sepsis-induced ALI/respiratory distress syndrome,aged 43-78 yr,weighing 49-89 kg,with their APACHE-Ⅱ scores of 16-23 and oxygenation index of 125-256 mm Hg,were randomly divided into 3 groups (n =20 each):conventional therapy group (group S),conventional therapy + acupuncture at non-acupoint group (group SNE) and conventional therapy + acupuncture at acupoint group (group SE).Bilateral Huantiao and Chize points were stimulated with electric stimulator (frequency 2/50 Hz,wave length 300μs,starting at a voltage of 0 and increasing by 0.1 mA every time until the maximal tolerance level was reached) for 30 min once a day for 5 days based on the conventional therapy in group SE.In group SNE,EA was performed at the non-acupoint based on the conventional therapy and the parameters of EA were same as those previously mentioned in group SE.Before EA stimulation (T1) and at 3 and 5 days after the end of EA stimulation (T2-3),arterial blood samples were taken for blood gas analysis,oxygenation index (OI) was calculated,and APACHE-Ⅱ] scores were assessed.Venous blood samples were collected at T1 and T3 for detection of TNF-α and IL-10 concentrations in the broncho-alveolar lavage fluid (BALF) and serum (by ELISA).Results Compared with the baseline value at T1,OI was significantly increased and APACHE-Ⅱ scores were decreased at T2 and T3,and TNF-α concentrations in the BALF and serum were decreased and IL-10 concentrations in the BALF and serum were increased at T3 in the three groups (P ＜ 0.05 or 0.01).The OI was significantly higher and APACHE-Ⅱ scores were lower at T3 than at T2 in the three groups (P ＜ 0.05).Compared with group S,the OI was significantly increased at T2 and T3,APACHE-Ⅱ scores and TNF-α concentrations in the BALF and serum were decreased at T3,and IL-10 concentrations in the BALF and serum were increased at T3 in group SE (P ＜0.05 or 0.01),and no significant changes were found in the parameters mentioned above in group SNE (P ＞0.05).There were no significant differences in the incidence and survival rate of multiple organ dysfunction syndrome among the three groups (P ＞ 0.05).Conclusion EA at Zusanli and Chize is helpful in mitigating sepsisinduced ALI in patients,and regulation of the balance between proinflammatory factors and anti-inflammatory factors and inhibition of inflammatory responses may be involved in the mechanism.

Effects of sevoflurane or ischemic preconditioning on the expression of ERK and CaM during lung ischemia-reperfusion in rats / 中华麻醉学杂志

Objectlve To investigate the effects of sevoflurane or ischemic preconditioning on the expression of extracellular signal-regulated kinase (ERK) and calmodulin (CaM) during lung ischemia-reperfusion (I/R)in rats. Methods Twenty-four male SD rats weighing 270-320 g were randomly divided into 4 groups ( n = 6each): sham operation group (group S), group I/R, ischemic preconditioning group (group IP), and sevoflurane preconditioning group (group SP). In group S, the hilum of the left lung was dissociated after thoracotomy but not occluded. In group I/R, lung I/R was produced by occlusion of the hilum of the left lung for 45 min followed by 120 min of reperfusion. In group IP, the hilum of the left lung was occluded for 5 min and unclamped for 5 min for 2 times before the model was established. In group SP, sevoflurane was inhaled for 30 min at the end-tidal concentration of 2.1% before lung ischemia. All rats were sacrificed at 120 min of reperfusion and the lung tissues were taken for determination of the contents of TNF-α and IL-6 by ELISA and the expression of ERK mRNA and CaM mRNA by RT-PCR. Results Compared with group S, the contents of TNF-α and IL-6 and the expression of ERK mRNA and CaM mRNA were significantly increased in group I/R, IP and SP ( P ＜ 0.05). Compared with group I/R, the contents of TNF-α and IL-6 and the CaM mRNA expression were significantly decreased, while the expression of ERK mRNA was significantly increased in group IP and SP ( P ＜ 0.05). There was no significant difference in the each index metioned above between group IP and SP ( P ＞ 0.05 ). Conclusion Sevoflurane preconditioning and ischemic preconditioning can protect the lung from I/R injury through down-regulating the expression of CaM and up-regulating the expression of ERK.

Preparation of the monoclonal antibody against SOX4 protein and detection of SOX4 expression level in different tumor cell lines / 生物工程学报

In the present study, we constructed a prokaryotic expression vector containing SOX4 protein encoding sequences. The GST-SOX4 soluble protein was expressed in Escherichia coli DH5alpha and purified by glutathione sepharose-4B. The purified recombinant protein was used to immunize Balb/C mice and the monoclonal antibody against SOX4 was prepared by using hybridoma technique. The titer of the antibody was determined as 1 x 10(5) by indirect ELISA. The specificity of the antibody was verified by Western blotting analysis. The monoclonal antibody specifically recognized the overexpressed exogenous SOX4 protein as well as endogenous SOX4 protein. The expression level of SOX4 protein in different cell lines and mouse tissues was detected by using the antibody. Differential expression of the protein was demonstrated by Western blotting. The data indicated that the antibody was specific. The antibody can be used as an important tool for further exploration of the role of SOX4 in tumorigenesis.

Preparation of monoclonal antibody against CUEDC2 protein and detection of expression profile of CUEDC2 in mouse tissues / 西安交通大学学报(医学版)

Objective To prepare monoclonal antibody(mAb) against CUE domain containing 2(CUEDC2) protein and detect the expression level of CUEDC2 in mouse tissues.Methods Fusion protein GST-human CUEDC2(hCUEDC2) was expressed and purified,and mAb against CUEDC2 was prepared by using hybridoma technique.The specificity of the antibody and CUEDC2 expression in mouse tissues were detected by Western blot analysis.Results We obtained mAb against CUEDC2 protein,which recognized specifically both hCUEDC2 and mouse CUEDC2.The mAb could be used to detect expression level of CUEDC2 protein,which was different in various mouse tissues.Conclusion The prepared CUEDC2 has good mAb specificity and can be used to detect expression of CUEDC2 protein in mouse tissue,thus providing a useful tool for further investigation into the functions of CUEDC2.