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ObjectiveTo study the differences in volatile oil content of bran-processed Atractylodes lancea and its standard decoction concentrate and freeze-dried powder， as well as the differences in the types and contents of chemical components in volatile oil， and to clarify the quality value transmitting. MethodTen batches of A. lancea rhizoma were collected and prepared into raw products and bran-processed products of A. lancea， standard decoction concentrate and freeze-dried powder of bran-processed A. lancea in order to extract the volatile oil， and the transfer rate of volatile oil in each sample was calculated. Quantitative analysis of the main chemical components（β-eudesmol， atractylon， atractylodin） in each volatile oil was performed by gas chromatography（GC） on the HP-5 quartz capillary column（0.32 mm×30 m， 0.25 μm） with a flame ionization detector（FID）， a split ratio of 10∶1 and a temperature program（initial temperature at 80 ℃， hold for 1 min， rise to 150 ℃ at 10 ℃·min-1， hold for 10 min， rise to 155 ℃ at 0.5 ℃·min-1， hold for 5 min， rise to 240 ℃ at 8.5 ℃·min-1， hold for 8 min）. Cluster analysis and principal component analysis（PCA） were used to explore the overall differences in types and contents of chemical components between the standard decoction concentrate and freeze-dried powder. ResultThe transfer rates of volatile oil in the bran-processed products， standard decoction concentrate and freeze-dried powder were 70.51%， 1.57% and 40.90%， respectively. The average transfer rates of β-eudesmol， atractylon and atractylodin in the volatile oil of bran-processed A. lancea were 58.45%， 48.49% and 55.64%， respectively. In the standard decoction concentrate， only β-eudesmol and atractylodin were detected， and their average transfer rates were 0.22% and 0.10%， respectively. And only β-eudesmol was detected in the freeze-dried powder with the average transfer rate of 8.37%. The results of cluster analysis and PCA showed that there are obvious differences in the types and contents of chemical components between the standard decoction concentrate and freeze-dried powder. ConclusionThe quality value transmitting between bran-processed A. lancea and its standard decoction concentrate and freeze-dried powder is stable， and if the freeze-dried powder is selected as the reference material of dispensing granules， appropriate amount of volatile oil should be added back to make it consistent with the quality of the standard decoction concentrate.
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ObjectiveTo explore the correlation between dryness and energy metabolism of Atractylodis Rhizoma， and to analyze the difference of medicinal properties between Atractylodes lancea and A. chinensis. MethodA total of 110 healthy SD rats were randomly divided into 11 groups， including normal group， volatile oil of A. lancea 1-5 group （S1-S5 group， doses of 447， 473， 442， 489， 496 mg·kg-1） and volatile oil of A. chinensis 1-5 group （N1-N5 group， doses of 197， 118， 281， 222， 185 mg·kg-1）， the administration volume was 0.01 mL·g-1 with intragastric administration for 21 days. Dryness effect of A. lancea and A. chinensis on rats was evaluated by comparing the body weight， drinking water volume， urine volume， whole blood viscosity and pathological sections of submandibular gland stained with hematoxylin-eosin （HE）. The expression of aquaporin 2 （AQP2） in rat kidney was measured by immunohistochemistry， the mRNA expressions of cytochrome C oxidase subunit 7A2 （COX7A2） and succinate dehydrogenase （SDH） complex subunit D （SDHD） in liver tissue were detected by real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. The contents of SDH， lactate dehydrogenase （LDH） and sodium ion-potassium ion-adenosine triphosphatase （Na+-K+-ATPase） in rat plasma were determined by colorimetry. The quality of A. lancea and A. chinensis was evaluated by coefficient of variation method， and Pearson correlation coefficient was used to analyze the correlation between dryness and energy metabolism. ResultCompared with the normal group， the amounts of drinking water and urine in volatile oil of A. lancea group and volatile oil of A. chinensis group increased， and the submandibular gland acini atrophied， the whole blood viscosity of rats in the volatile oil of A. lancea group increased significantly （P<0.01）， the expression levels of COX7A2 and SDHD mRNA， the activities of SDH， LDH and Na+-K+-ATPase increased significantly （P<0.01）， and the expression of AQP2 in kidney decreased significantly （P<0.01）. Compared with the normal group， the expression level of COX7A2 mRNA， SDH activity and whole blood viscosity in the volatile oil of A. chinensis group increased （P<0.05， P<0.01）， the AQP2 and SDH mRNA expression levels， LDH and Na+-K+-ATPase activities had no significant difference. The comprehensive score analysis of each index showed that the effect of volatile oil of A. lancea on dryness and energy metabolism was stronger than that of volatile oil of A. chinensis， and there was a positive correlation between dryness index and energy metabolism index. ConclusionThe two indexes show that medicinal properties of A. lancea is stronger than that of A. chinensis， and energy metabolism is closely related to the dryness of Atractylodis Rhizoma. It is suggested that it is reasonable to evaluate the dryness effect of Atractylodis Rhizoma from the perspective of energy metabolism， which can further enrich the evaluation indexes of medicinal properties.
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ObjectiveTo explore the correlation between dryness and energy metabolism of Atractylodis Rhizoma， and to analyze the difference of medicinal properties between Atractylodes lancea and A. chinensis. MethodA total of 110 healthy SD rats were randomly divided into 11 groups， including normal group， volatile oil of A. lancea 1-5 group （S1-S5 group， doses of 447， 473， 442， 489， 496 mg·kg-1） and volatile oil of A. chinensis 1-5 group （N1-N5 group， doses of 197， 118， 281， 222， 185 mg·kg-1）， the administration volume was 0.01 mL·g-1 with intragastric administration for 21 days. Dryness effect of A. lancea and A. chinensis on rats was evaluated by comparing the body weight， drinking water volume， urine volume， whole blood viscosity and pathological sections of submandibular gland stained with hematoxylin-eosin （HE）. The expression of aquaporin 2 （AQP2） in rat kidney was measured by immunohistochemistry， the mRNA expressions of cytochrome C oxidase subunit 7A2 （COX7A2） and succinate dehydrogenase （SDH） complex subunit D （SDHD） in liver tissue were detected by real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. The contents of SDH， lactate dehydrogenase （LDH） and sodium ion-potassium ion-adenosine triphosphatase （Na+-K+-ATPase） in rat plasma were determined by colorimetry. The quality of A. lancea and A. chinensis was evaluated by coefficient of variation method， and Pearson correlation coefficient was used to analyze the correlation between dryness and energy metabolism. ResultCompared with the normal group， the amounts of drinking water and urine in volatile oil of A. lancea group and volatile oil of A. chinensis group increased， and the submandibular gland acini atrophied， the whole blood viscosity of rats in the volatile oil of A. lancea group increased significantly （P<0.01）， the expression levels of COX7A2 and SDHD mRNA， the activities of SDH， LDH and Na+-K+-ATPase increased significantly （P<0.01）， and the expression of AQP2 in kidney decreased significantly （P<0.01）. Compared with the normal group， the expression level of COX7A2 mRNA， SDH activity and whole blood viscosity in the volatile oil of A. chinensis group increased （P<0.05， P<0.01）， the AQP2 and SDH mRNA expression levels， LDH and Na+-K+-ATPase activities had no significant difference. The comprehensive score analysis of each index showed that the effect of volatile oil of A. lancea on dryness and energy metabolism was stronger than that of volatile oil of A. chinensis， and there was a positive correlation between dryness index and energy metabolism index. ConclusionThe two indexes show that medicinal properties of A. lancea is stronger than that of A. chinensis， and energy metabolism is closely related to the dryness of Atractylodis Rhizoma. It is suggested that it is reasonable to evaluate the dryness effect of Atractylodis Rhizoma from the perspective of energy metabolism， which can further enrich the evaluation indexes of medicinal properties.
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Objective:To identify the key genes for neuropathic pain in rats.Methods:The genomic data of spinal cord tissues of rats (GSE18803) were downloaded from the Gene Expression Database at the American Center for Biotechnology Information to identify differentially expressed genes associated with neuropathic pain, and key genes were obtained by further analysis of the protein-protein interaction networks.Single-cell localization and expression of the key genes were analyzed by the Tabula Muris database.Results:The protein-protein interaction networks identified 10 hub genes, including Tyrobp, Clec4a3, C1qc, Ptprc, Laptm5, Csf1r, C1qa, C1qb, Fcgr3a, Cd53. Cd53, Laptm5 and Ptprc were mainly expressed in macrophages, B cells, NK cells, monocytes and granulocytes. Clec4a3 and Csf1r were mainly expressed in monocytes, Fcgr3a in monocytes and granulocytes, and Tyrobp in macrophages, monocytes, granulocytes, and pluripotent progenitor cells. Conclusions:Ten target genes associated with neuropathic pain are identified using bioinformatics, and their distribution and expression in immune inflammatory cells are obtained through comprehensive analysis.
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Objective:To investigate the efficacy of auricular acupoint bean-embedding therapy in the treatment of residual dizziness after successful reduction of benign paroxysmal positional vertigo (BPPV) and its effect on skin sympathetic response.Methods:A total of 110 patients with residual dizziness after successful reduction of BPPV who were admitted to Wenzhou Hospital of Traditional Chinese Medicine from January 2019 to May 2021 were included in this study. They were randomly divided into control and study groups, with 55 patients in each group. Patients in the control group were treated with drugs, and those in the study group were treated with auricular acupoint bean-embedding therapy. Before and after treatment, dizziness handicap inventory (DHI) score, activities-specific balance confidence (ABC) score, and Hamilton Anxiety Scale (HAMA) score were compared between the two groups. The latency and amplitude of sympathetic skin response (SSR) were measured.Results:After 15 days of treatment, DHI scores of all dimensions in the study group were significantly lower than those in the control group ( t = 16.13-20.62, all P < 0.05). ABC score in the study group was significantly higher than that in the control group [(87.90 ± 6.01) points vs. (80.55 ± 8.73) points, t = 3.10, P < 0.05). HAMA score in the study group was significantly lower than that in the control group [(7.85 ± 1.06) points vs. (13.30 ± 2.49) points, t = 8.43, P < 0.001]. The initial latency value of SSR in the study group was significantly higher than that in the control group [(1.95 ± 0.27) ms vs. (1.67 ± 0.21) ms, and the amplitude of SSR in the study group was significantly lower than that in the control group [(1.49 ± 0.15) mV vs. (1.70 ± 0.22) mV, t = 4.73, 4.04, both P < 0.001]. The incidence of adverse reactions in the study group was significantly lower than that in the control group [0.0% (0/55) vs. 10.9% (6/55), χ2 = 4.40, P < 0.05]. Conclusion:Auricular acupoint bean-embedding therapy can effectively alleviate the symptoms of dizziness after successful reduction of BPPV, improve patient's psychological status and autonomic nerve function, which is worthy of clinical promotion.
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Objective:To evaluate the role of nucleotide-binding oligomerization domain-2 (NOD2) in dorsal root ganglion in the development of neuropathic pain (NP) in rats.Methods:Thirty-two adult male SPF Sprague-Dawley rats, weighing 240-260 g, aged 2-3 months, were divided into 4 groups ( n=8 each) using a random number table method: control group (group C), NP group (group S), negative control siRAN group (group N), and NOD2-siRNA group (group R). In N and R groups, 1×10 8 IFU/ml negative control siRNA and NOD2-siRNA 10 μl were intrathecally injected, respectively, once a day for 3 consecutive days.Normal saline 10 μl was intrathecally injected once a day for 3 consecutive days in C and S groups.The model of NP was established using spared nerve injury (SNI) at 2 weeks after intrathecal injection.The mechanical paw withdrawal threshold (MWT) was measured at 1 day before surgery and 1, 3, 7, 10, 14 and 28 days after SNI.Animals were sacrificed after measuring pain threshold on day 28, and the dorsal root ganglions (DRGs) of the lumbar segment (L 4-6) were removed for determination of the expression of NOD2 (by Western blot) and expression of tumor necrosis factor-alpha (TNF-α), interleukin-1beta (IL-1β), IL-6 and NOD2 mRNA (using quantitative real-time polymerase chain reaction). Results:Compared with group C, MWT was significantly decreased at each time point after SNI, and the expression of NOD2 protein and mRNA and TNF-α, IL-1β and IL-6 mRNA in DRGs was up-regulated in group NP ( P<0.01). Compared with group NP, MWT was significantly increased at each time point after SNI, and the expression of NOD2 protein and mRNA and TNF-α, IL-1β and IL-6 mRNA in DRGs was down-regulated in group R ( P<0.01), and no significant change was found in the parameters mentioned above in group N ( P>0.05). Conclusion:The mechanism underlying the development of NP may be related to the up-regulation of NOD2 expression in DRGs, thus further promoting the expression of pro-inflammatory factors in rats.
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Objective To evaluate the relationship between the mechanism of spinal interleukin-17 (IL-17) regulating neuropathic pain (NP) and peripheral nerve-related proteins in rats. Methods Thirty-six healthy adult male Sprague-Dawley rats, in which IT catheters were successfully implanted, aged 10-12 weeks, weighing 240-260 g, were divided into 4 groups ( n=9 each) using a random number table method: sham operation group (Sham group), NP group, blank vector group (BV group), and IL-17 siRNA recombinant lentivirus group ( siRNA group) . NP was induced by chronic constriction injury ( CCI) to the sciatic nerve at 5 days after IT catheters were successfully implanted. At day 7 after CCI, normal sa-line 20 μl was intrathecally injected in Sham and NP groups, and in BV and siRNA groups, blank vector IL-17 siRNA-GFP and recombinant lentivirus 1 × 107 TU 10 μl were intrathecally injected, respectively, the catheter was flushed with normal saline 10 μl once a day for 4 consecutive days. The mechanical paw withdrawal threshold ( MWT) and thermal paw withdrawal latency ( TWL) were measured on day 1 before CCI and days 1, 7, 10, 11, 12, 13 and 14 after CCI. Proximal sciatic nerve tissues on the operated side were obtained at 14 days after CCI, the total protein was extracted, and the isobaric tags for relative and absolute quantification and liquid chromatography-tandem mass spectrometry were performed to stratify the differentially expressed proteins. Results Compared with group Sham, the MWT was significantly de-creased and TWL was shortened at each time point after CCI in the other three groups (P＜0. 05). Com-pared with NP and BV group, the MWT was significantly increased and TWL was prolonged at days 10-14 after CCI in group siRNA ( P＜0. 05) . There was no significant difference in the parameters mentioned above between group NP and group BV (P＞0. 05). Fifty-eight differentially expressed proteins were identified, and among the 58 proteins, the expression of 21 proteins was significantly up-regulated and the expression of 37 proteins was down-regulated. Conclusion Fifty-eight peripheral nerve-related proteins identified are related to the mechanism of spinal IL-17 regulating neuropathic pain in rats.
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Objective To evaluate the role of interleukin-17 (IL-17) in the peripheral nerve in blood-nerve barrier injury in a rat model of neuropathic pain (NP).Methods Forty healthy adult male Sprague-Dawley rats,weighing 250-280 g,were divided into 4 groups (n=10 each) using a random number table method:sham operation group (Sham group),NP group,blank vector group (BV group) and IL-17 siRNA recombinant lentivirus group (siRNA group).The rats were anesthetized with chloral hydrate,and intrathecal catheters were implanted.NP was produced by chronic constriction injury (CCI) to the sciatic nerve.Normal saline 10 μl was intrathecally injected at day 3 after CCI in Sham and NP groups.Blank vector and IL-17 siRNA-GFP recombinant lentivirus 1× 107 TU 10 μl were intrathecally injected,and then the tube was washed using normal saline 10 μl with the total volume of 20 μl once a day for 4 consecutive days.The mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured on day 1 before CCI and days 1,7,10,11,12,13 and 14 after CCI.Animals were sacrificed after the last behavior testing,and a segment of injured sciatic nerve 7 mm in length and containing 3 ligatures were harvested for examination of the ultrastructure and for detection of the expression of IL-17,occludin and claudin-5 by Western blot.Results Compared with Sham group,the MWT was significantly decreased,TWL was shortened,the expression of IL-17 was up-regulated,and the expression of occludin and claudin-5 was down-regulated at each time point after CCI in the other groups (P＜0.05 or 0.01).Compared with NP group or with BV group,the MWT was significantly increased,TWL was prolonged,the expression of IL-17 was down-regulated,the expression of occludin and claudin-5 was up-regulated (P＜0.05),and the pathological changes of sciatic nerve were significantly attenuated in siRNA group.Conclusion The mechanism of blood-nerve barrier injury may be related to increased level of IL-17 in the peripheral nerve in a rat model of NP.
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Objective To evaluate the changes in the expression of artemin in skin around the inci-sion during remifentanil-induced hyperalgesia in the rats with incisional pain. Methods Thirty-two healthy male Sprague-Dawley rats, aged 10-12 weeks, weighing 250-280 g, were divided into 4 groups (n = 8 each) using a random number table: control group (group C), incisional pain group (group I), remifen-tanil group (group R) and incisional pain plus remifentanil group (group I+R). Remifentanil was intrave-nously infused for 60 min at a rate of 1 μg·kg-1 ·min-1 in group R. In group I, the model of incisional pain was established, and the equal volume of normal saline was infused for 60 min via the tail vein at the same time. In group I+R, the model of incisional pain was established, and remifentanil was infused for 60 min via the tail vein at a rate of 1 μg·kg-1 ·min-1 at the same time. The equal volume of normal saline was infused for 60 min via the tail vein in group C. Mechanical paw withdrawal threshold (MWT) and ther-mal paw withdrawal latency (TWL) were measured at 24 h before infusion of remifentanil or normal saline and 2, 6, 24 and 48 h after the end of infusion (T0-4 ). Rats were sacrificed following the last measurement of pain threshold, and ipsilateral plantar skin was removed for detection of the expression of artemin protein and mRNA (by fluorescent quantitative real-time polymerase chain reaction or Western blot). Results Compared with group C, MWT was significantly decreased and TWL was shorten at T1-4 , and the expression of artemin protein and mRNA in plantar skin was up-regulated in R, I and I+R groups (P<0. 01). Compared with R and I groups, MWT was significantly decreased and TWL was shorten at T1-4 , and the ex-pression of artemin protein and mRNA in plantar skin was up-regulated in group I+R (P<0. 01). Conclu-sion The peripheral mechanism by which remifentanil induces hyperalgesia may be related to up-regulated expression of artemin in skin around the incision in the rats with incisional pain.
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Objective To evaluate the role of D-serine in nerve cell apoptosis induced by multiple exposures to sevoflurane anesthesia in newborn mice and its relationship with glycogen synthase kinase-3 beta (GSK-3β).Methods Thirty healthy male C57B/L6 mice,aged 6 days,weighing 3.5-4.5 g,were divided into 3 groups (n =10 each) using a random number table:control group (group C),multiple exposures to sevoflurane anesthesia group (group S) and D-serine group (group D).On postnatal days 6,7 and 8,3％ sevoflurane in 30％ oxygen was inhaled for 2 h starting from 10:00 daily,and normal saline 0.1 ml and D-serine 500 mg/kg were intraperitoneally injected at 30 min before inhalation in S and D groups,respectively.In group C,30％ oxygen was inhaled for 2 h starting from 10:00 daily,and normal saline 0.1 ml was intraperitoneally injected at 30 min before inhalation.The animals were sacrificed after the end of oxygen or sevoflurane inhalation on postnatal day 8,and the brains were removed for determination of the expression of phosphorylated GSK-3β (pGSK-3β) and activated caspase-3 in brain tissues by Western blot.Results Compared with group C,the expression of pGSK-3β in brain tissues was significantly down-regulated,and the expression of activated caspase-3 in brain tissues was up-regulated in group S (P＜ 0.05),and no significant change was found in the parameters mentioned above in group D (P＞0.05).Compared with group S,the expression of pGSK-3β in brain tissues was significantly up-regulated,and the expression of activated caspase-3 in brain tissues was down-regulated in group D (P＜0.05).Conclusion D-serine is involved in the nerve cell apoptosis induced by multiple exposures to sevoflurane anesthesia through inhibiting the activation of GSK-3β in newborn mice.
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Objective To evaluate the effect of dexmedetomidine on necroptosis during liver injury in septic rats.Methods Eighteen SPF adult male Sprague-Dawley rats,weighing 200-220 g,were divided into 3 groups (n=6 each) using a random number table:sham operation group (group SH),sepsis group (group SEP) and dexmedetomidine group (group DEX).Sepsis was induced by cecal ligation and puncture in chloral hydrate-anesthetized rats in SEP and DEX groups.Dexmedetomidine 5 μg/kg was injected via the caudal vein at 1 h before operation in group DEX.Blood samples were collected from the caudal vein at 6 h after operation for determination of serum aspartate amino-transferase (AST) and alanine aminotransferase (ALT) concentrations.The rats were then sacrificed and livers were removed for determination of the level of reactive oxygen species (ROS) in liver tissues (using chemiluminescence assay) and expression of receptor-interacting protein 1 (RIP1),RIP3,mixed lineage kinase domain-like (MLKL),high-mobility group box 1 protein (HMGB1) and dynamin-related protein 1 (Drpl) in liver tissues (by Western blot).Results Compared with group SH,the serum AST and ALT concentrations were significantly increased,the expression of RIP1,RIP3,MLKL,HMGB1 and Drpl in liver tissues was up-regulated,and the level of ROS in liver tissues was increased in SEP and DEX groups (P＜0.05).Compared with group SEP,the serum AST and ALT concentrations were significantly decreased,the expression of RIP1,RIP3,MLKL,HMGB1 and Drp1 in liver tissues was down-regulated,and the level of ROS in liver tissues was decreased in group DEX (P＜0.05).Conclusion The mechanism by which dexmedetomidine attenuates liver injury may be related to inhibition of necroptosis in septic rats.
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Objective To evaluate the relationship between cold hyperalgesia and trafficking of transient receptor potential melastatin 8 (TRPM8) to cell membrane in the dorsal root ganglion (DRG) of rats with neuropathic pain (NP).Methods Ninety-six healthy male Sprague-Dawley rats,aged 10-12 weeks,weighing 250-280 g,were divided into sham operation group (S group,n=48) and NP group (n =48) using a random number table.NP was produced by chronic constriction injury to the sciatic nerve.The number of paw lifts on the cold plate and mechanical paw withdrawal threshold (MWT) were measured on 1 day before operation and 1,4,7,10 and 14 days after operation.Rats were sacrificed after behavioral testing,and ipsilateral DRGs of the lumbar segment (L46) were dissected tor detection of the expression of TRPM8 in total and membrane proteins by Western blot,and the ratio of TRPM8 expression in the membrane protein to that in the total protein (m/t ratio) was calculated.Results Compared with group S,the number of paw lifts on the cold plate was significantly increased,the MWT was decreased,the expression of TRPM8 in total and membrane proteins was up-regulated,and m/t ratio was increased on postoperative days 4,7,10 and 14 in group NP (P＜0.05 or 0.01).In group NP,the number of paw lifts on the cold plate was gradually increased with the prolongation of time after operation and reached the peak on postoperative day 10,maintaining at the peak until postoperative day 14;the MWT was gradually decreased and reached the lowest level on postoperative day 10,maintaining at the lowest level until postoperative day 14;the expression of TRPM8 in total and membrane proteins and m/t ratio were gradually increased with the prolongation of time after operation and reached the peak on postoperative day 10,maintaining at the peak level until postoperative day 14 (P＜0.01).Conclusion The mechanism underlying the development of cold hyperalgesia is related to enhanced trafficking of TRPM8 to cell membrane in DRGs of rats with NP.
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Objective To evaluate the changes in the expression of transient receptor potential vanilloid 1 (TRPV1) in dorsal root ganglions (DRGs) during remifentanil-induced hyperalgesia in the rats with incisional pain.Methods Thirty-two SPF healthy male Sprague-Dawley rats,weighing 240-260 g,aged 2-3 months,in which caudal catheters were successfully implanted,were divided into 4 groups (n=8 each) using a random number table:control group (group C),incisional pain group (group Ⅰ),remifentanil group (group R),and incisional pain + remifentanil group (group I+R).A 1 cm longitudinal incision was made through skin,fascia and muscle of the plantar aspect of the left hindpaw to establish the model of incisional pain.In group R,remifentanil was intravenously infused for 60 min at a rate of 1.2 μg · kg-1 · min-1.In group Ⅰ,the model of incisional pain was established,and the equal volume of normal saline was intravenously infused for 60 min at the same time.In group I+R,the model of incisional pain was established,and remifentanil was intravenously infused for 60 min at a rate of 1.2 μg · kg-1 · min-1 at the same time.In group C,the equal volume of normal saline was intravenously infused for 60 min.The mechanical paw withdrawal threshold (MWT) and thermal paw withdrawl latency (TWL) were measured at 24 h before normal saline or remifentanil infusion (To) and 2,6,24 and 48 h after the end of infusion (T1-4).The rats were sacrificed after the last measurement of pain threshold,and the DRGs of the lumbar segment (L4-6) were removed for determination of the expression of TRPV1 protein and mRNA by Western blot and real-time polymerase chain reaction,respectively.Results Compared with group C,the MWT was significantly decreased,and the TWL was shortened at T1-4,and the expression of TRPV1 protein and mRNA was up-regulated in R,I and I+R groups (P＜0.05).Compared with group R or group I,the MWT was significantly decreased,and the TWL was shortened at T1-4,and the expression of TRPV1 protein and mRNA was up-regulated in group I+R (P＜0.05) Conclusion Up-regulated expression of TRPV1 in DRGs may be involved in the mechanism underlying remifentanil-induced hyperalgesia in the rats with incisional pain.
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Objective To evaluate the efficacy of different low-doses of nalmefene in preventing remifentanil-induced postoperative hyperalgesia. Methods One hundred American Society of Anesthesiolo-gist physical status Ⅰor Ⅱpatients, aged 20-64 yr, wih body mass index of 18-25 kg∕m2, scheduled for elective gynecological laparoscopic surgery under general anesthesia, were divided into 4 groups(n=25 each)using a random number table: control group(group C)and different doses of nalmefene groups (N1, N2 and N3 groups). In N1, N2 and N3 groups, nalmefene 02, 03 and 05 μg∕kg(diluted to 5 ml in normal saline)were intravenously injected, respectively, at 5 min before anesthesia induction, while the equal volume of normal saline was given in group C. Anesthesia was induced with midazolam 005 mg∕kg, sufentanil 03 μg∕kg, etomidate 03 mg∕kg and rocuronium 06 mg∕kg. The patients were me-chanically ventilated after tracheal intubation. Anesthesia was maintained by IV infusion of remifentanil 03 μg·kg-1·min-1and inhalation of 4%-6% desflurane, bispectral index value was maintained at 45-60, and muscle relaxation was maintained with intermittent IV boluses of rocuronium. After admission to postan-esthesia care unit, patient-controlled analgesia(PCA)was performed, and PCA solution contained sufen-tanil 1 μg∕ml in 100 ml of normal saline. PCA pump was programmed to deliver a 05 ml bolus dose with a lockout interval of 15 min and background infusion at 2 ml∕h. Numeric rating scale score was maintained <4. The time for remifentanil infusion was recorded. The consumption of sufentanil was recorded in 0-1, 1-3, 3-6, 6-12 and 12-24 h periods after surgery, and the occurrence of nausea, vomiting, tachycardia, hypertension and shivering was also recorded within 24 h after surgery. Results Compared with group C, the postoperative consumption of sufentanil was significantly reduced in 0-1 h and 1-3 h periods after sur-gery in group N1 and in 0-1, 1-3, 3-6 and 6-12 h periods after surgery in group N2, and the incidence of postoperative nausea was significantly decreased in N1, N2 and N3 groups(P<005). The consumption of sufentanil in 3-6 h period after surgery was significantly lower in group N2 than in group N1(P<005). Conclusion The optimal dose of nalmefene is 03 μg∕kg when used to prevent remifentanil-induced post-operative hyperalgesia.
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Objective To evaluate the role of glial cell line-derived neurotrophic factor family re-ceptor alpha3(GFRα3)in the expression and membrane trafficking of transient receptor potential melasta-tin 8(TRPM8)in the dorsal root ganglion(DRG)during cold hyperalgesia in rats with neuropathic pain (NP). Methods Thirty-two healthy adult male Sprague-Dawley rats, aged 10-12 weeks, weighing 250-280 g, in which intrathecal catheters were successfully implanted, were divided into 4 groups(n=8 each) using a random number table: sham operation plus GFRα3 dsRNA group(Sham+dsRNA group), sham operation plus GFRα3 siRNA group(group Sham+siRNA), NP plus GFRα3 dsRNA group(group NP+dsRNA)and NP plus GFRα3 siRNA group(group NP+siRNA). NP was produced by chronic constriction injury to the sciatic nerve. At 10-30 days after operation, GFRα3 dsRNA 10 μg∕20 μl was intrathecally injected once a day for 4 consecutive days in Sham+dsRNA and NP+dsRNA groups, and 10 μg∕20 μl GFRα3 siRNA, of which the sense strand was modified with 2′-O-methyl and 5′-cholesterol, was intrathe-cally injected once a day for 4 consecutive days in Sham+siRNA and NP+siRNA groups. The number of paw lifts on the cold plate, mechanical paw withdrawal threshold(MWT)and thermal paw withdrawal latency (TWL)were measured on 1 day before operation and 10, 11, 12, 13(before intrathecal injection)and 14 days after operation. The rats were sacrificed after the last behavioral testing, and ipsilateral DRGs of the lumbar segment(L4-6)were dissected for detection of the expression of GFRα3 and TRPM8 in total and membrane proteins by Western blot, and the ratio of TRPM8 expression in the membrane protein to that in the total protein(m∕t ratio)was calculated. Results Compared with group Sham+dsRNA, the number of paw lifts on the cold plate was significantly increased, the MWT was decreased, and TWL was shortened after operation in NP+dsRNA and NP+siRNA groups, the expression of GFRα3 and TRPM8 in total and membrane proteins was significantly up-regulated, and m∕t ratio was increased in group NP+dsRNA, and the expression of GFRα3 in DRGs was significantly down-regulated(P<001), and no significant change was found in the expression of TRPM8 in total and membrane proteins or m∕t ratio in group NP+siRNA(P>005). Compared with group NP+dsRNA, the number of paw lifts on the cold plate was significantly de-creased, the expression of GFRα3 and TRPM8 in total and membrane proteins was down-regulated, m∕t ra-tio was decreased(P<001), and no significant change was found in MWT or TWL in group NP+siRNA (P>005). Conclusion GFRα3 in DRGs can up-regulate the expression of TRPM8 and enhance the membrane trafficking of TRPM8, which may be involved in the maintenance mechanism of cold hyperalgesia in rats with NP.
ABSTRACT
Objective To investigate the expression of the Kv3 .4 protein and Kv3 .4 mRNA in various stages of oral carcino‐genesis .Methods The expression of Kv3 .4 protein and Kv3 .4 mRNA were detected by immunohistochemistry (SP method) and RT‐PCR technique respectively in the oral carcinogenesis of SD rat which were induced by 4NQO .Results With the aggravation of the epithelial dysplasia ,Kv3 .4 protein and Kv3 .4 mRNA expression increased gradually .They were strongly positive in oral squa‐mous cell carcinoma .Conclusion The expression of Kv3 .4 protein and Kv3 .4 mRNA levels increased consistently with the aggra‐vation of the epithelial dysplasia .
ABSTRACT
Objective:To investigate the expression of HERG1 and Kv3.4 in normal oral mucosa(NOM)and oral lichen planus (OLP).Methods:20 OLP and 1 6 NOM specimens were collected and immunohistochemically stained(IHC)by SP method for the detection of HERG1 and Kv3.4 protein expression.Results:HERG1 and Kv3.4 were negative or weak-positive in the sapmles of bas-al layer of NOMand non-erosive OLP,but positive in basal layer,spinous layer and granular layer of erosive OLP.The expression of HERG1 and Kv3.4 was higher in OLP than in NOM tissues (P <0.05);and higher in erosive OLP than in non-erosive OLP(P <0.05).In OLP HERG1 expression was positively related to Kv3.4(P <0.05).Conclusion:HERG1 and Kv3.4 may be related to the development of OLP.