Activation of A1 reactive astrocytes in the medullary dorsal horn of rats participates in the chronification of trigeminal neuralgia / 中南大学学报(医学版)

Objective:The activation of astrocytes is an important process in the formation of chronic pain.This study aims to observe the activation of A1 reactive astrocytes in the medullary dorsal horn in the rat model of trigeminal neuralgia,and to explore the mechanism of central sensitization caused by A1 reactive astrocyte. Methods:The adult male rats were randomly divided into a sham group and a chronic constriction injury of infraorbital nerve(ION-CCI)group.The facial mechanical pain threshold and thermal withdrawal latency were measured before the operation and on the 1st,3rd,7th,10th,and 14th day after the operation.After pain behavior observation,the expression of glial fibrillary acidic protein(GFAP)in the medullary dorsal horn was observed by immunohistochemistry and immunofluorescence colocalization of GFAP,complement 3(C3)/S100A10,and 4',6-diamidino-2-phenylindole(DAPI)was analyzed.Primary astrocytes were cultured and randomly divided into a naive group and a DHK group.The DHK group was treated with 1 mmol/L of astrocyte activation inhibitor dihydrokainic acid(DHK).Fura-2/AM was used to stain the astrocytes and the calcium wave of the 2 groups under the stimulation of high potassium was recorded and compared.The expression of C3 was detected by Western blotting. Results:The facial mechanical pain threshold and thermal withdrawal latency of the ION-CCI group were significantly lower than those of the sham group(both P<0.05).There were a large number of GFAP positive astrocytes in the medullary dorsal horn of the ION-CCI group.The fluorescence intensity of GFAP in the ION-CCI group was higher than that in the sham group(P<0.05).GFAP and C3/S100A10 were co-expressed in astrocytes.Compared with the sham group,the fluorescence intensity of C3 and the protein expression of C3 in the ION-CCI group were increased(both P<0.05).The expression of C3 in ION-CCI group was significantly increased(P<0.05).Compared with the naive group,the C3 protein expression was significantly decreased in the DHK group(P<0.05).The intensity of calcium fluorescence was increased after high potassium stimulation in both groups.Furthermore,the peak and increase amplitude of calcium fluorescence in the naive group were much higher than those in the DHK group(both P<0.05). Conclusion:A1 reactive astrocytes in the medullary dorsal horn of trigeminal neuralgia model rats are increased significantly,which may participate in central sensitization of trigeminal neuralgia by impacting astrocyte calcium wave.

Establishment and application of family-centred health education programe for family caregivers of Autistic children / 现代临床护理

Objective To propose a family-centred health education program for family caregivers of Autistic children and investigate its clinical value.Methods Delphi method was used to establish a family-centred Autistic children care program for family caregivers looking after the Autistic children,which involved two rounds of expert consultation.The pre-and post-control method for different cases was employed in the study.Forty Autistic caregivers in our hospital from January 2022 to October 2022 were assigned to the control group and another 40 in our hospital from November 2022 to January 2023 were assigned to the observation group.The family caregivers of Autistic children in the control group received traditional care education,while those in the observation group was managed with the family-centred Autistic children health education program.The two groups were compared in terms of care burden,knowledge of health education,and evaluation of clinical symptom of Autistic children.Results The expert authority coefficient of the first round of expert consultation was 0.875 respectively and those in the second round was 0.900,respectively.The coefficient of variation of the coordination degree of each index was 0.04-0.20.Kendalls W scores of the two rounds were 0.794 and 0.786,respectively.A health education program for family-centred Autistic children caregiver was established to consist of three subjects:caregiver training,family-centred parental care and activation of positive emotions of autistic caregivers.Autism caregivers in both groups had completed the study.In comparison with the control group,the care burden of the observation group was significantly lower,the knowledge of health education was significantly higher,and the evaluation of children's clinical symptoms was better among the family caregivers of autistic children in the observation group(all P<0.05).Conclusions The family-centred autistic children care program for family caregivers is scientific and applicable.It is conducive to providing health education guidance for family caregivers of Autistic children,effectively reducing the care burden and enhancing the knowledge of health education.

Effect of intrathecal exosomes derived from human amniotic fluid on neuropathic pain in mice / 中华麻醉学杂志

Objective:To evaluate the effect of intrathecal exosomes derived from human amniotic fluid (hAF exo) on neuropathic pain induced by spared nerve injury (SNI) in mice.Methods:Eighteen clean-grade healthy male Kunming mice, aged 7-8 weeks, weighing 30-35 g, were divided into 3 groups ( n=6 each) using a random number table method: sham operation group (Sham group), SNI group, and SNI+ hAF exo group. Spared nerve injury was produced by exposing the sciatic nerve and its branches and ligation and transection of tibial nerve and common fibular nerve in anesthetized mice. Another three mice were selected to develop the model of neuropathic pain after anesthesia. PKH-26 labeled hAF exo 7 μl was intrathecally injected on days 1, 2 and 3 after developing the model. The mice were sacrificed at 10 h after the end of administration, and the uptake of hAF exo by the dorsal horn of the injured lumbar enlargement of the spinal cord was observed with the fluorescence microscope. On 1, 2 and 3 days after developing the model, 1 μg/μl hAF exo 7 μl was intrathecally injected in SNI+ hAF exo group, and PBS 7 μl was intrathecally injected in Sham group and SNI group. The mechanical paw withdrawal threshold (MPWT) was measured at 1 day before and 1, 3, 5 and 7 days after operation. And then the mice were sacrificed after measurement of the pain threshold at 7 days after developing the model, and the ipsilateral lumbar enlargement of the spinal cord was taken for determination of the expression of CD11b, interleukin-1beta (IL-1β) and IL-10 by Western blot. Results:The dorsal horn of the lumbar enlargement of the spinal cord on the injured side could absorb hAF exo with the fluorescence microscope. Compared with Sham group, the MPWT was significantly decreased at 3-7 days after developing the model, the expression of CD11b and IL-1β was up-regulated ( P<0.05), and no significant change was found in the expression of IL-10 in SNI group ( P>0.05). Compared with SNI group, the MPWT was significantly increased at 3-7 days after developing the model, the expression of CD11b and IL-1β was down-regulated, and the expression of IL-10 was up-regulated in SNI+ hAF exo group ( P<0.05). Conclusions:Intrathecal exosomes derived from human amniotic fluid can alleviate neuropathic pain in mice, and the mechanism may be related to mediation of the polarization of microglia from M1 type to M2 type and attenuation of neuroinflammation.

Current situation and countermeasures of acute postoperative pain management / 中国医师杂志

Pain caused by surgery is an important clinical issue that seriously affects postoperative rehabilitation and health-related quality of life. Failure to effectively manage postoperative pain not only leads to a decrease in patient quality of life, increases medical expenses, but also has a negative impact on patient recovery. Therefore, it is of great clinical significance to address the challenges of acute postoperative pain management, find effective management strategies, and improve the quality of pain management. This article summarizes the current status of acute postoperative pain management in recent years, including the mechanism of pain occurrence, pain assessment methods, drug and non drug management strategies, and predictive factors for chronic postoperative pain. It also looks forward to future research directions and application prospects.

Comparison of perioperative multimodal analgesic regimens for patients with trigeminal neuralgia undergoing microvascular decompression surgery / 中国医师杂志

Objective:To explore a multimodal perioperative analgesia plan for patients undergoing microvascular decompression surgery for trigeminal neuralgia.Methods:Eighty patients who underwent microvascular decompression surgery for trigeminal neuralgia admitted to the Xiangya Hospital, Central South University from April 2017 to April 2019 were randomly divided into a nerve block group (group A) and a control group (group C) using a random number table method, with 40 patients in each group. The group A underwent surgical block of the lateral occipital and auricular nerves under ultrasound guidance before induction, with 3 ml of 0.5% ropivacaine used at each site. The group C did not undergo nerve block. Both groups received intravenous injections of midazolam, sufentanil, cisatracurium, etomidate, and lidocaine for anesthesia induction, followed by tracheal intubation and maintenance of anesthesia with propofol and remifentanil. After surgery, an analgesic pump was connected. The total amount of intraoperative use of sufentanil and remifentanil in both groups was recorded, as well as the pain Visual Analogue Scale (VAS) and postoperative anesthesia related complications at 2, 6, 24, and 48 hours after surgery.Resultsl:The total amount of sufentanil and remifentanil used during surgery in the group A was less than that in the group C (all P<0.05). The incidence of postoperative nausea and vomiting in the group A patients was lower than that in the group C ( P<0.05), and the nausea and vomiting score was also lower than that in the group C ( P<0.05). There was no statistically significant difference in the incidence of other postoperative complications (all P>0.05). There was a statistically significant difference in VAS scores between the two groups at 6 hours after surgery ( P<0.05). Conclusions:Occipital and auricular nerve blockade can reduce the amount of opioid drugs used during microvascular decompression surgery in patients with trigeminal neuralgia, thereby reducing the incidence of nausea and vomiting. The postoperative analgesic effect is good.

The effect of oxycodone on postoperative acute pain in patients undergoing thoracoscopic surgery / 中国医师杂志

Objective:To retrospectively observe the effect of oxycodone on acute postoperative pain in patients undergoing thoracoscopic surgery.Methods:A retrospective analysis was conducted on the clinical data of 404 patients undergoing thoracoscopic thoracic surgery under combined general anesthesia at the Xiangya Hospital, Central South University from April 1, 2020 to September 30, 2021. They were divided into A group ( n=99, oxycodone group) and B group ( n=305, control group) based on whether oxycodone was used during the surgery. The two groups of patients were further matched 1∶1 using the nearest neighbor matching method. We compared the Visual Analogue Scale (VAS) of activity and resting pain and the incidence of moderate to severe pain between two groups of patients 24 hours after surgery, and observed the incidence and severity of pain related adverse reactions such as nausea, vomiting, itching, and dizziness. Resultsl:After matching the propensity scores of the two groups of patients, the balance was good ( SMD<0.20). There was no statistically significant difference between the groups in age, gender, body mass index, American Society of Anesthesiologist (ASA) grade, surgical time, intraoperative bleeding, and the use of antiemetics and analgesics during the perioperative period (all P>0.05). Compared with the control group, patients in the group A had a resting VAS [(2.03±1.61)points vs (1.62±1.31)points, P=0.049], and activity VAS [(4.13±1.72)points vs (3.51±1.79)points, P=0.013] was even lower, and the incidence of moderate to severe pain (VAS≥4) during activity was lower [59.6%(59/99) vs 37.4%(37/99), P=0.003]. There was no statistically significant difference in the incidence of analgesic related adverse reactions between the two groups ( P>0.05). Conclusions:Intravenous injection of oxycodone can effectively alleviate acute pain in patients undergoing thoracoscopic surgery within 24 hours, and reduce the incidence of moderate to severe pain during activity.

Berberine targets the electron transport chain complex I and reveals the landscape of OXPHOS dependency in acute myeloid leukemia with IDH1 mutation / 中国天然药物

Metabolic reprogramming, a newly recognized trait of tumor biology, is an intensively studied prospect for oncology medicines. For numerous tumors and cancer cell subpopulations, oxidative phosphorylation (OXPHOS) is essential for their biosynthetic and bioenergetic functions. Cancer cells with mutations in isocitrate dehydrogenase 1 (IDH1) exhibit differentiation arrest, epigenetic and transcriptional reprogramming, and sensitivity to mitochondrial OXPHOS inhibitors. In this study, we report that berberine, which is widely used in China to treat intestinal infections, acted solely at the mitochondrial electron transport chain (ETC) complex I, and that its association with IDH1 mutant inhibitor (IDH1mi) AG-120 decreased mitochondrial activity and enhanced antileukemic effect in vitro andin vivo. Our study gives a scientific rationale for the therapy of IDH1 mutant acute myeloid leukemia (AML) patients using combinatory mitochondrial targeted medicines, particularly those who are resistant to or relapsing from IDH1mi.

Effects of intrathecal injection of ropivacaine on spinal cord in rats based on genomics / 中南大学学报(医学版)

OBJECTIVES@#There are clinical reports of nerve injury caused by ropivacaine. The mechanism for nerve injury induced by ropivacaine has not been fully clarified. This study aims to investigate the changes of pain threshold and L3 spinal cord genomics at 6 h and 24 h after intrathecal injection of 0.5% and 1.0% ropivacaine, and to explore the underlying mechanisms for nerve injury caused by ropivacaine.@*METHODS@#A total of 30 male Sprague Dawley rats weighing 220-260 g were successfully implanted with microspinal catheter. The rats were randomly divided into 5 groups (each n=6): a control group (given saline), a ropivacaine group 1 and a ropivacaine group 2 (both given 1% ropivacaine), a ropivacaine group 3 and a ropivacaine group 4 (both given 0.5% ropivacaine). The rats received continuous intrathecal injection of corresponding drugs at 8.3 μL/h for 24 h via an implanted intrathecal catheter followed by 24 h-pause of injection for the ropivacaine group 2, the ropivacaine group 4 and the control group, 6 h-pause of injection for the ropivacaine group 1 and the ropivacaine group 3. For each group, the observation of behavioral change and the paw withdrawal mechanical threshold (PWMT) was conducted immediately after the injection and again after the pause of injection. After the PWMT observation, the rats were dissected to acquire L3 spinal cords. Illumina sequencing was applied to construct gene libraries. Then the statistical methods were used to find out differentially expressed genes between the groups. Gene Ontology (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) signaling pathway analysis were conducted for those genes. Real-time RT-PCR was used to determine different expressions of some of those genes.@*RESULTS@#Compared with control group, the PWMT got higher in the ropivacaine group 1-4 and was positively correlated with concentration, negatively correlated with discontinuation duration. Compared with control group, the ropivacaine group 1 had 488 differentially expressed genes, of which 456 were up-regulated and 32 were down-regulated; the ropivacaine group 2 had 1 194 differentially expressed genes, of which 1 092 were up-regulated and 102 were down-regulated; the ropivacaine group 3 had 518 differentially expressed genes, of which 384 were up-regulated and 134 were down-regulated; and the ropivacaine group 4 had 68 differentially expressed genes, of which 46 were up-regulated and 22 were down-regulated. GO enrichment analysis and KEGG signaling pathway analysis showed that most of these differentially expressed genes were related to signaling pathways of inflammatory response.@*CONCLUSIONS@#After intrathecal injection of 0.5% ropivacaine and 1.0% ropivacaine for 24 h, the differentially expressed genes in L3 spinal cord of rats are mainly related to signaling pathways of inflammatory response.

The concept of enhanced recovery after surgery improves upon quality of anesthesia management / 中国医师杂志

Enhanced recovery after surgery (ERAS) is mainly based on evidence-based medicine to implement a series of measures to optimize perioperative management, reduce patients′ physiological and psychological trauma stress during perioperative period, reduce patients′ functional damage and promote patients′ functional recovery, so as to achieve rapid rehabilitation. ERAS has been widely used in clinic and achieved good clinical results. However, it still faces a series of problems that need further research to clarify the clinical path of ERAS required by different patients and different surgical methods. In particular, it is necessary to strengthen the research on risk factors of ERAS, minimally invasive surgery, goal-directed fluid therapy, anesthesia and postoperative pain management technology, pre rehabilitation and postoperative rehabilitation technology, and implement ERAS guided by the best outcome in the perioperative period.

The application of enhanced recovery after surgery in perioperative management of thyroid ambulatory surgery / 中国医师杂志

Objective:Through retrospective analysis of perioperative management data of ambulatory thyroid surgery under the concept of enhanced recovery after surgery (ERAS), we provide foundation for the safe implementation of ambulatory thyroid surgery.Methods:From January 2019 to December 2019, patients undergoing thyroid surgery were enrolled in the study under the concept of ERAS at the ambulatory surgery center of Xiangya Hospital, Central South University. Data of patients during perioperative period were collected, including adverse events, anesthesia recovery, postoperative and post-discharge recovery were recorded.Results:This study was included 703 cases of patients, thyroid nodules in 374 cases, thyroid malignant tumor in 329 cases. There were no significance difference in the operation time, anesthesia time, wake up of time, and postanesthesia care unit (PACU) time between the two groups (all P>0.05). No hypertension, hypotension, tachycardia, bradycardia or other arrhythmias occurred during perioperative period. No adverse events such as intraoperative awareness and delay of wake up occurred. No severe pain, nausea, vomiting, dizziness and other discomfort occurred after surgery. All 703 patients were discharged from hospital within 24 hours. Conclusions:Anesthesiologists participate in patient management according to the perioperative medicine requirements, and ambulatory thyroid surgery may be performed safely under the concept of ERAS.

Research progress on predicting consciousness recovery during recovery period of general anesthesia patients / 中国医师杂志

It is necessary to use objective and accurate methods to assess the changes of the consciousness of patients emergencing from general anesthesia. In this way, adverse medications during the waking period can be avoided, and it can ensure the stable and safe recovery of consciousness of the patients, quickly remove the adverse factors affecting the patients, and strive to reduce the occurrence of complications during the waking period. This article briefly reviews the research progress of bispectral index and other common clinical anesthesia depth monitoring techniques used to assess the changes of consciousness of patients awakening from general anesthesia, and explores the regular pattern of recovery of consciousness in patients awakening from general anesthesia, in order to reduce complications in the recovery period .

Role of SPP1 in endogenous protective mechanism underlying neuropathic pain in mice with spinal cord injury: relationship with VEGF-AKT signaling pathway / 中华麻醉学杂志

Objective:To evaluate the role of secreted phosphoprotein 1 (SPP1) in endogenous protective mechanism underlying neuropathic pain (NP) in mice with spinal cord injury and the relationship with the vascular endothelial growth factor (VEGF)/kinase B (Akt) signaling pathway.Methods:Seventy-two clean-grade healthy female Kunming mice, aged 7-8 weeks, weighing 30-35 g, were divided into 4 groups ( n=18 each) using a random number table method: sham group (Sham group), NP caused by spinal cord injury group (NP group), NP caused by spinal cord injury+ SPP1-siRNA group (NS-siRNA group), and NP caused by spinal cord injury+ adeno-associated virus vector group (NP-EV group). A model of NP was established by a semi-transecting of the spinal cord.AAV-SPP1-siRNA-GFP adenovirus and AAV-vector-GFP adenovirus 7 μl were intrathecally injected in NS-siRNA group and NP-EV group, respectively, and 5 days later the model was established.At 1, 2 and 3 weeks after operation, 6 mice in each group were randomly selected to measure paw withdrawal threshold to mechanical stimulation (PWMT) and tail flick latency (TFL) to thermal stimuli.And then the mice were sacrificed and the ipsilateral injured spinal cord tissues were taken for determination of the expression of SPP1 mRNA (by real-time polymerase chain reaction) and expression of SPP1, VEGF, Akt and phosphorylated Akt (p-Akt) (by Western blot). Results:Compared with group Sham, PWMT was significantly decreased, TFL was shortened, and the expression of SPP1 mRNA, SPP1, VEGF and p-Akt protein was up-regulated at 1, 2 and 3 weeks after operation in NP, NS-siRNA and NP-EV groups( P<0.05). Compared with group NP, PWMT was significantly decreased, TFL was shortened, and the expression of SPP1 mRNA, SPP1, VEGF and p-Akt protein was down-regulated at 1, 2 and 3 weeks after operation in group NS-siRNA( P<0.05). Compared with group NS-siRNA, PWMT was significantly increased, TFL was prolonged, and the expression of SPP1 mRNA, SPP1, VEGF and p-Akt protein was up-regulated at 1, 2 and 3 weeks after operation in group NS-siRNA( P<0.05). Conclusion:SPP1 is involved in the endogenous protective mechanism underlying NP in mice with spinal cord injury, which may be related to the activation of the VEGF/AKT signaling pathway.

Research on the application of multiple interactive modes in online teaching: taking Anesthesia Equipment teaching as an example / 中华医学教育探索杂志

This research focuses on the application of multiple interactive modes in online teaching, combined with the actual teaching cases of the anesthesia equipment course of Xiangya Anesthesiology Specialty of Central South University, showing in detail the preparations for interactive teaching before anesthesia equipment learning, the interaction in online classrooms, the extension of interactive teaching outside the classroom, and the evaluation of interactive teaching feedback mechanism throughout the implementation process. By establishing a "host-guest-viewer" mode, the effect of online live broadcasting is maximized. Through the 360-degree materialized explanation with students as the main body, we will make opening in the pain points and blocking points of online teaching in which students do not go to class and students have no thinking, and promote the improvement of online teaching quality and efficiency. In the following practice, we must continue to work on issues such as the improvement of teacher talent quality, the building of an efficient talent team, and the construction of practical application value evaluation systems for teaching.

Practice of situational case teaching in online teaching of anesthesiology internship under the COVID-19 epidemic / 中华医学教育探索杂志

Objective:To explore the application and effect of the situational case teaching in online internship teaching of anesthesiology.Methods:Thirty-four anesthesiology undergraduates in batch 2015 were randomized into two groups, the experimental group and the control group, with 17 students in each group. The experimental group adopted situational case teaching method, and the control group were taught by the traditional case-based teaching method. The questionnaire survey was used to assess the evaluation of students' personal comprehensive ability and teaching effect on the two groups. The data were analyzed by SPSS 19.0, and the measurement data were expressed by (mean ± standard deviation). Independent sample t test was used for comparison between the two groups, with significant difference when P<0.05. Results:The experimental group is superior than the control group in improvement of learning interest, self-learning ability, comprehensive expression and, communication ability, problem analyzing and solving ability, ability of uniting theory with practice, adaptability, and teamwork ability ( P<0.05). The recognition degree of this teaching mod and teaching effects is also significantly higher than that of the control group ( P<0.05). Conclusion:The situational case teaching mode has achieved good results in the online internship teaching of anesthesiology. This method can stimulate students' learning interest, improve learning efficiency, enhance students' ability of clinical practice and comprehensive quality, and also be beneficial to the improvement of teaching quality.

Perioperative management for patients with coronavirus disease 2019 / 中南大学学报(医学版)

The coronavirus disease 2019 (COVID-19) is a new infectious disease, which has a strong virus transmission power and complex transmission routes. This disease is prone to outbreak of cluster infection. It is difficult for medical workers to provide a better perioperative treatment for surgery patient with COVID-19 while avoiding hospital spread effectively. The perioperative management for such patients needs to fully consider the possible lung injury factors caused by anesthesia and surgery. It also needs to choose the suitable timing of the operation, carry out preoperative infection screening and evaluation, and implement lung protection strategies during and after the operation to avoid aggravating the lung injury. Meanwhile, it is necessary to pay more attention to infection prevention and control in order to avoid nosocomial infection.

Strengthen application research of ultrasound technology in perioperative and pain medicine / 中国医师杂志

The development of portable ultrasound equipment and point-of-care ultrasonography has promoted the application of ultrasound technology in the field of anesthesia and pain medicine.ultrasound is primarily used to evaluate blood volume and volume responsiveness,airway,lung and stomach volume,as imaging guidance tools for regional anesthesia and chronic pain management in perioperative and pain clinics.Transesophageal echocardiography is mainly used to obtain the information of cardiac anatomy,pathology and cardiac function.It is especially advantageous for the unsatisfactory image obtained by transthoracic ultrasound,left auricular thrombosis,infective endocarditis,aortic dissection,intraoperative monitoring,etc.The application of ultrasound technology improves the accuracy and safety of perioperative and pain medical diagnosis and treatment,and improves the quality of perioperative and pain medical treatment.

Role of long non-coding RNA MEG3 in hyperglycose-induced neurocyte damage in rats:relation-ship with mitochondrion-dependent apoptosis / 中华麻醉学杂志

Objective To evaluate the role of long non-coding RNA maternally expressed gene 3(MEG3)in hyperglycose-induced neurocyte damage and the relationship with mitochondrion-dependent ap-optosis in rats.Methods Normally cultured PC12 cells were divided into 5 groups(n＝18 each)using a random number table method: normal concentration of glucose control group(C group),normal concentra-tion of glucose plus MEG3 group(C+MEG3 group),high-concentration glucose group(HG group),high-concentration glucose plus MEG3 group(HG+MEG3 group),and high-concentration glucose plus negative lentiviral vector(LV-NC)group(HG+NC group).PC12 cells were cultured in DMEM medium with 25 mmol/L glucose in group C.PC12 cells were cultured in DMEM medium with 25 mmol/L glucose after being transfected with MEG3 lentiviral vector(LV-MEG3)in C+MEG3 group.PC12 cells were cultured in DMEM medium with 250 mmol/L glucose in HG group.PC12 cells were incubated in DMEM medium con-taining 250 mmol/L glucose after being transfected with LV-MEG3 in HG+MEG3 group.PC12 cells were in-cubated in DMEM medium containing 250 mmol/L glucose after being transfected with LV-NC in HG+NC group.After the cells were cultured or incubated for 1 day,the cell viability was measured by CCK8 assay,the apoptosis rate and level of reactive oxygen species(ROS)were determined by flow cytometry,and the amount of lactic dehydrogenase(LDH)released was measured by DCFH-DA,the expression of Cyt c,caspase-3,caspase-9,Bcl-2,Bax and Apaf-1 was determined by Western blot,and the opening of mito-chondrial permeability transition pore(mPTP)was determined by fluorescent method.Blc-2/Bax ratio was calculated.Results Compared with group C,the cell viability was significantly decreased,the amount of LDH released,ROS level and apoptosis rate were increased,the opening of mPTP was increased,and the expression of caspase-3,caspase-9,Cyt c,Bax,Bcl-2 and Apaf-1 was up-regulated in HG,HG+MEG3 and HG+NC groups,and Bcl-2/Bax ratio was increased in HG+MEG3 group and decreased in HG and HG+NC groups(P＜0.05).Compared with HG group and HG+NC group,the cell activity was significantly in-creased,the amount of LDH released,ROS level and apoptosis rate were decreased,the opening of mPTP was decreased,the expression of caspase-3,caspase-9,Cyt c,Bax,and Apaf-1 was down-regulated,the expression of Bcl-2 was up-regulated,and Bcl-2/Bax ratio was increased in HG+MEG3 group(P＜0.01).Conclusion MEG3 may be involved in the endogenous protective mechanism during hyperglycose-induced neurocyte damage by inhibiting mitochondrion-dependent apoptosis in rats.

Aucubin alleviates lipopolysaccharide-induced acute lung injury in mice / 中南大学学报(医学版)

To investigate the effect of prophylactic aucubin (AU) on lipopolysaccharide (LPS)-induced acute lung injury (ALI) in mice. 
 Methods: Male BABL/c mice were randomly divided into a control group, an ALI group, and an AU treatment group, 16 mice in each group. ALI mice were injected with LPS (5 mg/kg, intratracheal injection), and AU (10 mg/kg) was injected intraperitoneally 30 min ahead. After LPS injection for 6 hours mice were sacrificed, the morphological changes of lung tissues were detected by HE staining and the lung injury score was obtained. The mRNA expression of tumor necrosis factor-α (TNF-α) and interleukin 10 (IL-10) in lung tissue was detected by real-time PCR. The total protein and lactate dehydrogenase (LDH) activity, the cell count, and the protein content of TNF-α and IL-10 in the mouse bronchoalveolar lavage fluid (BALF) were detected.
 Results: Compared with ALI mice, the pathological damage score of lung tissue was significantly reduced in the AU group, the total number of BALF cells, neutrophils, and macrophages were significantly decreased, LDH activity and the total protein content were also significantly decreased (all P<0.01). In addition, AU can reduce the mRNA and protein expression of TNF-α in lung of ALI mice, and increase the mRNA and protein expression of IL-10 (all P<0.01).
 Conclusion: AU can reduce LPS-induced ALI in mice.

Establishment and differential protein identification of two-dimensional gel electrophoresis for proteomics in the spinal cord of morphine-tolerant rats / 中南大学学报(医学版)

To establish a two-dimensional gel electrophoresis (2-DE) map for comparative proteomic analysis of rat spinal cord with chronic morphine tolerance, and to detect differentially expression proteins that are associated with chronic morphine tolerance.
 Methods: Sixteen male SD rats received the intrathecal catheterization operation and they were randomly divided into a morphine tolerance group (MT group, n=8) and a saline group (NS group, n=8). The lumbar enlargement segments of the MT group and the NS group spinal cord were harvested and proteins were separated by 2-DE. Differential proteome profiles were established and analyzed by means of immobilized pH gradient-based two-dimensional polyacrylamide gel electrophoresis (2D-PAGE). The 2-DE maps were visualized after coomassie blue staining and analyzed using PDQuest analysis software. Identification of differential protein spots was conducted by MALDI-TOF-MS, and the Mascot query software was used to search Swiss-Prot database for bioinformatics analysis. Western blotting was used to verify the expression of some differentially expressed proteins.
 Results: A total of 1 000 spots were identified in 2-DE maps of rat spinal cord tissues from the MT group and the NS group, and 36 proteins were significantly differentially expressed in the MT group compared with the NS group. Identification was conducted by MALDI-TOF-MS and Swiss-Prot database through Mascot query software, and a total of 14 proteins were obtained. Among them, 2 protein spots were down-regulated in the MT group compared with that in the NS group, and 12 protein spots were up-regulated in the MT group compared with that in the NS group. Two kinds of proteins (NUDAA, ENOG) were verified by Western blotting and the results were consistent with proteomics data.
 Conclusion: The optimized 2-DE profiles for the proteome of spinal cord tissue in rats with chronic morphine tolerance is established preliminarily, which showed that morphine tolerance can cause changes in the expression of various proteins in the spinal cord.

mTOR signaling pathway of spinal cord is involved in peripheral nerve injury-induced hyperalgesia in rats / 中南大学学报(医学版)

To investigate whether mammalian target of rapamycin (mTOR) signaling pathway is involved in peripheral nerve injury-induced hyperalgesia through activation of spinal dorsal astrocytes in rats.
 Methods: A total of 30 male Sprague-Dawley (SD) rats were randomly divided into 6 groups (n=5): the 1 day group (D1 group), the 4 days group (D4 group), the 7 days group (D7 group), the 14 days group (D14 group), the normal group and the sham group. The sciatic nerve chronic constriction injury (CCI) model was established in the D1, D4, D7 and D14 group. The normal group received no treatment while the sham group was only exposed the sciatic nerve. Paw withdrawal mechanical threshold (PWMT) and paw withdrawal thermal latency (PWTL) were measured at the 1st, 4th, 7th, and 14th day after CCI in the different groups. Lumbar spinal cord were harvested on the 1st, 4th, 7th and 14th day in the D1, D4, D7, D14 group correspondingly, which were harvested on the 14th day in the normal group and the sham group. Distribution of mTOR in rat spinal cord was assessed by immunohistochemistry. The expressions of mTOR mRNA and protein in the spinal cord in different groups were determined by real-time PCR and Western blotting, respectively. Another 30 male intrathecal catheterized SD rats were randomly divided into 6 groups (n=5): a blank group, a CCI group, a CCI+early rapamycin (RAPA) group, a CCI+early dimethylsulfoxide (DMSO) group, a CCI+ later RAPA group, and a CCI+later DMSO group. The blank group didn't received any treatment; The CCI group was carried out the treatment of CCI model in the left hind limbs. 10 μL of 1% RAPA was given to the CCI+early RAPA group intrathecally at 4 hours after CCI for 3 days; the CCI+later RAPA group were treated with the same dose of RAPA on the 7th days after CCI for 3 days; the CCI+early DMSO group and the CCI+later DMSO group were injected with the same volume of 4% DMSO at the corresponding time as controls. The PWTL and PWMT were measured before and after intrathecal catheterization, and every other day after CCI. The lumbar spinal cords were selected and the expression of glial fibrillary acidic protein (GFAP) in spinal dorsal horn were examined by immunohistochemistry in the 14th day after CCI.
 Results: The immunohistochemistry positive particles of mTOR were widely distributed in the cytoplasm of the normal spinal neurons. Compared with the base line, the PWMT in the D14 group on the 1st, 4th, 7th and 14th day after CCI were significantly lower, and the PWTL on the 4th, 7th and 14th day after CCI were also significantly lower (P<0.05 or P<0.01). The expressions of mTOR mRNA and protein in the CCI groups (D1, D4, D7 and D14 group) were significantly increased than those in the normal group (P<0.05 or P<0.01). Compared with the CCI+early DMSO group, the PWMT and PWTL in the CCI+early RAPA group were obviously increased on 4th, 6th, 8th, 10th, 12th or 14th day after CCI (P<0.05 or P<0.01); compared with the CCI+later DMSO group, the PWMT and PWTL in the CCI+later RAPA group were also significantly increased at the 8th, 10th or 14th day after CCI (P<0.01 or P<0.05). The GFAP immunohistochemistry positive area and absorbance value in the dorsal horn of the lumbar spinal cord in the CCI rats were decreased in the CCI+early RAPA group compared with the CCI+early DMSO group (P<0.05 or P<0.01), and which were also decreased in the CCI+later RAPA group compared with the CCI+later DMSO group (P<0.05 or P<0.01).
 Conclusion: mTOR signaling pathway may be involved in hyperalgesia induced by peripheral nerve injury via spinal astrocyte activation in the dorsal horn of the spinal cord.