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.

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.

P38 MAPK inhibitor SB203580 attenuates the toxicity of ropivacaine on PC12 cells / 中南大学学报(医学版)

To investigate the effect of SB203580, a p38MAPK specific inhibitor, on ropivacaine-induced cytotoxicity in PC12 cells.
 Methods: PC12 cells were divided into three groups: the normal group (Group N), cells were cultured for 48 h; the ropivacaine group (Group R), cells were cultured with 15 mmol/L ropivacaine hydrochloride for 48 h; the ropivacaine+SB203580 group (Group R+S), cells were cultured with 15 mmol/L ropivacaine hydrochloride plus 10 μmol/L SB203580 for 48 h. The cell survival rates were detected by MTT assay. The protein levels of cleaved caspase-3, phosphor-p38 (p-p38) and cystolic cytochrome C (Cyt C) were detected by Western blotting.
 Results: Compared with the Group N, the number and survival rate of PC12 cells in the Group R and the Group R+S were significantly reduced (all P<0.05); the number and survival rate of PC12 cells in the Group R+S were significantly higher than those in the Group R (both P<0.05). Compared with the Group N, the levels of p-p38 and cleaved caspase-3, and the content of cytoplasmic Cyt C in the PC12 cells from the Group R and the Group R+S were significantly enhanced (all P<0.05); compared with the Group R, the levels of p-p38 and cleaved caspase-3, and the content of cytoplasmic Cyt C in the PC12 cells from the Group R+S were decreased (all P<0.05).
 Conclusion: The ropivacaine-induced cytotoxicity can be attenuated via inhibition of p38MAPK; which is related to decrease in Cyt C content and cleaved caspase-3 expression.

Vascular endothelial growth factor antibody attenuates diabetic peripheral neuropathic pain in rats / 中南大学学报(医学版)

To explore the role of vascular endothelial growth factor (VEGF) in diabetic peripheral neuropathic pain in rats.
 Methods: Twenty-four adult male Sprague-Dawley rats aged 8 weeks were randomly divided into 3 groups (n=8 per group). The control group (C group): rats were intraperitoneally injected with sodium citrate solution at 10 mL/kg; the model group (M group): rats were intraperitoneally injected with streptozotocin at 65 mg/kg; the treatment group (T group): rats received intraperitoneal injection of anti-VEGF antibody (10 mg/kg) at the 1st, 3rd, 7th, 10th day after STZ treatment. Meanwhile, rats of C and M group were received with the same volume of sodium citrate solution. Blood glucose was measured before 1 day or at the 1st, 3rd, 7th or 14th day after receiving STZ. Body weight, paw withdrawal mechanical threshold (PWMT) and paw withdrawal thermal latency (PWTL) were measured before 1 day or at the 1st, 3rd, 5th, 7th, 10th or 14th day after receiving STZ. All lumbar spinal cords were dissected to examine the p-protein kinase B (p-Akt) and transient receptor potential vanilloid 1 (TRPV1) expression by Western blot.
 Results: After injection with STZ, the body weight showed significant differences at some time point between the M, T or C group (P<0.01); body weight of rat in the C group were increased gradually. Compared with the C group, the fast blood glucose in the M or the T Group at the same time points were increased significantly (P<0.01). The PWMT and PWTL of the M, T or C group were significant difference among various time points (P<0.01). The PWMT and PWTL in the M or T group were obviously reduced compared with those in the C group (P<0.01). Compared with the M group, the PWMT and PWTL in the T group were increased at the 10th or 14th day (P<0.01 or P<0.05). Compared with the C group, the p-Akt and TRPV1 levels in the M and T group were increased (P<0.01). Compared with the M group, p-Akt and TRPV1 levels in T group were decreased (P<0.01).
 Conclusion: VEGF is able to regulate the expression of TRPV1 through PI3K/Akt pathway, which contributes to diabetic peripheral neuropathic pain in rats. Anti-VEGF treatment may be useful for alleviation of diabetic peripheral neuropathic pain.

Effect of neurotrophin-3 on Akt expression during ropivacaine-induced neurotoxicity to spinal cord of rats / 中华麻醉学杂志

Objective To evaluate the effect of neurotrophin-3 (NT-3) on the expression of serine/threonine protein kinase (Akt) during ropivacaine-induced neurotoxicity to the spinal cord of rats.Methods Healthy adult male Sprague-Dawley rats,aged 1-2 months,weighing 280-320 g,were used in the study.A catheter was inserted at L5,6 interspace into the epidural space of rats.A total of 108 rats,in which intrathecal catheters were successfully implanted,were randomly divided into 3 groups (n =36each):control group (group C),1％ ropivacaine group (group R),and 1％ ropivacaine + NT-3 group (group NT).The equal volume of normal saline was given in group C,1％ ropivacaine 0.12 ml/kg was injected via the intrathecal catheter once every 1.5 h for 8 times in total in R and NT groups.In addition,NT-3 0.1 mg/kg was simultanenously injected via the intrathecal catheter in group NT.On days 1,3,5,7,14 and 28 after the end of administration (T1-6),6 rats were sacrificed in each group.Their lumbar enlargements were removed for determination of neuronal apoptosis (using TUNEL) and Akt expression (by immuno-histochemistry).The apoptotic rate was calculated.Results Compared with group C,the apoptotic rate was significantly increased at T1-4,and Akt expression was significantly up-regulated at T1-3 in group R,and the apoptotic rate were significantly increased,and Akt expression was significantly up-regulated at T1-3 in group NT (P＜0.05).Compared with group R,the apoptotic rate was significantly decreased at T3,4,and Akt expression was significantly down-regulated at T2.3 in group NT (P＜0.05).Conclusion The mechanism by which NT-3 reduces ropivacaine-induced neurotoxicity to the spinal cord may be related to down-regulation of the expression of Akt in rats.

Efficacy of different methods of anesthesia on children underwent hypospadias surgery / 中南大学学报(医学版)

OBJECTIVE@#To assess the efficacy of different methods of anesthesia on children underwent hypospadias surgery.
@*METHODS@#A total of 90 children (2-6 years old, 11.5-21.0 kg weight) with I or II grade of hypospadias based on ASA standard, who scheduled for hypospadias angioplasty, were randomly divided into 3 groups: Group I, general anesthesia combined epidural anesthesia; Group II, laryngeal mask airway under general anesthesia; Group III, laryngeal mask airway under general anesthesia combined epidural block. All children were inhaled sevoflurane to keep bispectral index value in a range from 45 and 60. The Group I adopted epidural anesthesia after intravenous induction of anesthesia; the Group II was inserted laryngeal mask after induction; the Group III was inserted laryngeal mask after induction and adopted epidural block. The anesthesia efficacy, hemodynamic changes, adverse reaction and the postoperative complications were observed in the 3 groups.
@*RESULTS@#Compared with the Group I or the Group II, the blood pressure and heart rate ran more smoothly in the Group III, and the postoperative agitation and incidence of adverse events were also significantly reduced (all P<0.05).
@*CONCLUSION@#The laryngeal mask airway under general anesthesia combined epidural block is a better choice for children scheduled for hypospadias angioplasty.

Protective effects of pretreatment with neurotrophin-3 on intrathecal ropivacaine in rats / 中南大学学报(医学版)

OBJECTIVE@#To investigate the effect of pretreatment with neurotrophin-3 (NT-3) on intrathecal ropivacaine in rats.@*METHODS@#A total of 144 male Sprague Dawley rats weighing 280-320 g were successfully implanted with microspinal cather following the improved methods of Yaksh. The rats were randomly divided into 4 groups and given saline (Group NS, n=36), 0.5% ropivacaine (Group M, n=36), 1% ropivacaine (Group R, n=36), and ropivacaine+NT-3 (Group T, n=36). The rats received 0.12 mL/ kg body weight of ropivacaine at 0.5% or 1%, or normal saline only, via an implanted intrathecal catheter at 90-min interval for 12 h in Group NS, M, R and T. In the meantime the rats also received NT-3 0.1 mg/kg in group T. On days 1, 3, 5, 7, 14 and 28, we assessed the paw withdrawal mechanical threshold (PWMT) and paw withdrawal thermal latency (PWTL), behavioural change and histopathological damage score changed for possible neuronal injury within the spinal cord.@*RESULTS@#Compared with Group NS and Group M, the PWMT and PWTL were significantly higher on 1, 3, 5 d and the histopathological damage score was significantly higher on 1, 3, 5, 7, 14 d in Group R (P<0.05). Compared with Group T, the PWMT and PWTL in Group R were significantly higher on 1, 3, 5 d and histopathological damage score was significantly higher on 5, 7, 14 d (P<0.05).@*CONCLUSION@#NT-3 pretreatment in mice has obvious protective effect against repeated intrathecal injection of 1% ropivacaine in the spinal nerve.

Effect of lentiviral vector-mediated GDNF up-regulation on neuropathic pain of chronic constriction injury rats / 中国医师杂志

Objective To investigate the effect of intrathecal injection of lentiviral vector-mediated up-regulation of glial cell line-derived neurotrophicfactor (GDNF) on neuropathic pain of chronic constriction injury (CCI) rats.Methods The CCI model was prepared by ligating the sciatic nerve of Sprague-Dawley (SD) rats.Seven days after CCI modeling,a single intrathecal injection of lentiviral vectors (LV)-GDNF was given.Before CCI and 3,5,7,14,and 21 days after CCI modeling,the mechanical pain threshold was tested in rats,and 21 days after surgery,Western blot was used to detect the expression of GDNF protein.Results On 21 days after CCI modeling,GDNF expression was reduced compared to sham group.After intrathecal injection of LV-GDNF,GDNF expression was up-regulated in the spinal cord,and CCI-induced mechanical hyperalgesia in rats was alleviated.Conclusions Intrathecal injection LV-GDNF can up-regulate the expression of GDNF and alleviate neuropathic pain in CCI rats.

Changes in expression of purinergic P2X4 receptor in spinal cord dorsal horn and dorsal root ganglion in a rat model of acute morphine tolerance or inflammatory pain / 中华麻醉学杂志

ObjectiveTo evaluate the changes in the expression of purinergic P2X4 receptor (P2X4R) in spinal cord dorsal horn and dorsal root ganglion in a rat model of acute morphine tolerance or inflammatory pain.MethodsThirty male SD rats were randomly divided into 3 groups:normal saline group (group NS,n =5);acute morphine tolerance group (group M,n =5) and inflammatory pain group (group F,n =20).Inflammatory pain was induced by subcutaneous injection of 4％ formalin 50 μl into the plantar surface of left hindpaw in group F.The animals received intrathecal morphine 10 μg ( 10 μl) once every 2 h for 6 times (T1-6) in group M,while the equal volume of normal saline was given instead in group NS.The rats were then sacrificed 2 h after the last time administration.Paw withdrawal latency (PWL) to a thermal nociceptive stimulus was measured at T1-6 in groups M and NS,or on day 4,7,10 and 14 after establishing the model of inflammatory pain in group F,The rats were sacrificed after measurement of PWL and spinal cord dorsal horn and dorsal root ganglion were removed to detect the expression of P2X4 R by immuno-histochemisty.ResultsCompared with the baseline value,PWL was significantly decreased on day 4-14 after intlammatory pain in group F,and PWL was significantly increased at T1-5,while no significant change was found at T6 in group M ( P ＞ 0.05).Compared with group NS,the expression of P2X4 R was up-regulated in group M,and the expression of P2X4 R was up-regulated on day 4 after inflammatory pain,peaked on day 7 after inflammatory pain,and then was down-regulated gradually on day 10 and 14 after inflammatory pain in group F ( P ＜ 0.01).ConclusionThe expression of purinergic P2Y4 R is up-regulated in spinal cord dorsal horn and dorsal root ganglion in rats with acute morphine tolerance or inflammatory pain,and the change may be related to the development of acute morphine tolerance or inflammatory pain.

Incidence and risk factors of intraoperative awareness during general anesthesia / 中南大学学报(医学版)

OBJECTIVE@#To investigate the incidence of awareness during general anesthesia and analyze the risk factors in anesthetic practice and patient populations.@*METHODS@#A total of 2 300 patients who underwent general anesthesia were included. Perioperative data and anesthetic drugs were collected prospectively. Patients were interviewed twice postoperatively with the same structured questionnaire. Each patient was classified into categories as no awareness, possible awareness, and awareness.@*RESULTS@#Twenty-one patients (0.91%) definitely reported awareness, and another 205 (8.91%) reported possible awareness. Few of the patients with awareness required psychological intervention. ASA physical status III-IV and propofol maintenance were associated risk factors of awareness.@*CONCLUSION@#The incidence of intraoperative awareness is high in the clinical practice in major medical centers.

Construction of lentiviral vector of RNA interference of PKCY gene / 中华麻醉学杂志

Objective To construct a lentiviral vector of RNA interference (RNAi) of PKCγ gene. Methods The effective sequence of siRNA targeting PKCγ gene was confirmed in our previous study. The complementary DNA containing both sense and antisense oligo DNA of the targeting sequence was designed, synthesized and cloned into the pGCSIL-GFP vector, which contained U_6 promoter and green fluorescent protein (GFP) . The resulting lentiviral vector containing PKCγshRNA was named lentivinis RNAi vector of PKCγ, and it was confirmed by realtime PCR and sequencing. 293T cells were cotransfected with lentiviral vector pGCSIL-CTP, pHelper 1.0 and pHelper 2.0. All virus stocks were produced by calcium phosphate-mediated transfection. The titer of virus was tested according to the expression level of GFP. Results PCR and DNA sequencing demonstrated that the lentivirus RNAi vector of PKCγ producing PKCγshRNA was constructed successfully. The titer of concentrated virus was 1 ×10~9 TU/ml. Conclusion The lentivinis RNAi vector of PKCy was constructed successfully.

Effect of lentivirus-mediated RNA interference on expression of / 中华麻醉学杂志

Objective To investigate the effect of lentivirus-mediated RNA interference(RNAi) on the expression of PKCγ mRNA and protein in rat neurons.Methods Primary cultured cortical neurons from SD rat (16 days of pregnency) embryos were randomly divided into 3 groups with 6 wells in each group:control group (group C),negative group(group NC)and RNAi group.Group C received no treatment.Each well in group NC was given negative lentivirus 3 × 105.Each well in group RNAi was given the recombinant lentiviral vector containing PKCγ shRNA(LV-PKC7 shRNA).The expression of PKCT mRNA and protein in rat neurons was detected by RT-PCR and Western blot 5 days later.The interference efficiency was ealculated.Results Compared with group NC,the expression of PKCγ mRNA and protein was down-regulatedin group RNAi(P<0.05),but no significant change Was found in group NC(P>0.05).The interference efficiency of gene and protein were 99.3%and 85.2%respectively.Conclusion Lentivirus-mediated RNAi can down restate the expression of PKC7 gene and protein in rat neurons.