OBJECTIVE: To investigate the expression of neuronal nitric oxide synthase (nNOS) in spinal dorsal horn and the effect of intrathecal katemine on the expression of nNOS in the rats with formalin-induced pain. METHODS: Thirty-two Sprague-Dawley rats were randomly divided into 4 groups (n=8 in each group): a control group (C), an intrathecal saline group (NS), a katemine 50 microg group (K1), and a katemine 100 microg group (K2). The rats that were anesthetized with 10% chlroral hydrate 300 - 350 mg/kg by abdominal injection were intrathecally inserted a microspinal catheter into the lumbar region according to the method of modified Yaksh. After 5 days ,the rats in Group NS, K1 and K2 were intrathecal 20 microL saline or 10 microL katemine (50, 100 microg) followed by 10 microL saline, respectively. Thirty minutes later, 5% formalin 50 microL was subcutaneously injected into the left hindpaw. Pain intensity scoring (PIS) was used to assess antinociceptive behavior within 1 h after the formalin injection. The expression of nNOS in the spinal dorsal horn of L5 segment was assayed using immunohistochemistry 24 h later. RESULTS: Compared with Group NS, PIS of Group K1 and K2 was decreased obviously (P<0.01) in the second phase of formalin pain. The number of immunoreactive soma and immunohistochemistic dying grade of nNOS in the spinal dorsal horn of L5 segment was higher in Group NS than that in Group C (P<0.01), but Group K1 and K2 were lower than Group NS (P<0.01). CONCLUSION There was significant antinociception of intrathecal katemine in rats with formalin pain. Intrathecal katemine significantly inhibited the increase of nNOS expression in the spinal dorsal horn of formalin-induced pain, suggesting nNOS plays an important role in nociceptive transmission and modulation of the spinal cord.
Analgesics ; administration & dosage ; Animals ; Formaldehyde ; Injections, Spinal ; Ketamine ; administration & dosage ; Male ; Nitric Oxide Synthase Type I ; biosynthesis ; genetics ; Pain ; chemically induced ; drug therapy ; metabolism ; Posterior Horn Cells ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo establish a simple high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS) system for determination of capsaicin in rat plasma.

METHODSRat plasma (100 µl) was extracted with mixed solvent (ethyl acetate:acetone 85:15) after addition of 50 ng/ml internal standard, and analyzed by Agilent 6460 Triple Quadrupole LC/MS system. The detection was performed with multiple reactions monitoring using electrospray ionization. The precursor/product ion transitions were monitored at m/z 306→137 (+Ion mode) for capsaicin. Verapamil was used as the internal standard at m/z455→165.

RESULTSThe analysis time was only 1.5 min in the positive mode; the calibration curve was linear in the concentration range of 1.85-370 ng/ml. The lowest limit of quantification reached 1.85 ng/ml. The extraction recoveries were 77.34%, 70.64% and 78.02% for the three quality control concentration levels (3.7, 37, and 370 ng/ml), respectively.

CONCLUSIONThe LC-MS/MS system is simple, accurate, reliable and time-saving for determination of trace amount of capsaicin in plasma.

Animals ; Capsaicin ; blood ; Chromatography, High Pressure Liquid ; Plasma ; chemistry ; Rats ; Tandem Mass Spectrometry

OBJECTIVE: To determine an optimal clinical dose of ketamine after comparing the efficacy and security of 3 low dose ketamines mixed with butorphanol in the postoperative continuous intravenous analgesia. METHODS: Eighty ASA (American Society of Anesthesiologists) I-II patients scheduled for elective gynecological surgery under general anesthesia were divided randomly into 4 groups (n=20): Group B received butorphanol 3 microg/(kg x h);Group BK1 received butorphanol 2 microg/(kg x h) mixed with ketamine 60 microg/(kg x h); Group BK2 received butorphanol 2 microg/(kg x h) mixed with ketamine 90 microg/(kg.h); and Group BK3 received butorphanol 2 microg/(kg x h) mixed with ketamine 120 microg/(kg x h). Continuous intravenous infusion pump was used when the patients had obvious pain (visual analgesia scale of five), and the bolus infusion (4 mL) was given before the operation, and continuous infusion at 2 mL/h. In the postoperative period, pain was assessed using visual analogue scale (VAS) at 2,6,12,24, and 48 h.At the same time, Ramsay scores and adverse effects were recorded. RESULTS: There was no significant difference in the adverse effects and the postoperative mean arterial pressure, heart rate, respiratory rate values, and pulse oxygen among the 4 groups. Postoperative VAS values in Group BK3 was the lowest, followed by Group BK2. There was no significant difference between Group BK1 and Group B. The incidence of somnolence in Group B was higher than that in Group BK1, BK2 and BK3(P<0.05). CONCLUSION Ketamine in subanaesthetic dose added to butorphanol for postoperative continuous intravenous infusion has a better postoperative analgesic effect and sedation. It can effectively spare butorphanol consumption without increasing adverse effects. The optimal combined dose is 90-120 microg/(kg x h).
Adult ; Analgesia ; methods ; Analgesics ; administration & dosage ; Butorphanol ; administration & dosage ; Dose-Response Relationship, Drug ; Drug Therapy, Combination ; Female ; Gynecologic Surgical Procedures ; Humans ; Infusions, Intravenous ; Ketamine ; administration & dosage ; Pain, Postoperative ; drug therapy

The plateau pika (Ochotona curzoniae) and plateau zokor (Myospalax baileyi) are specialized native species of the Qinghai-Tibetan plateau. The goal of this study was to examine physiological differences in skeletal muscle glycolysis and hepatic lactate metabolism between these two species. The partial sequence of pyruvate carboxylase (PC) gene was cloned and sequenced. The mRNA expression levels of PC and lactate dehydrogenases (LDH-A, LDH-B) were determined by real-time PCR. The enzymatic activity of PC was measured using malic acid coupling method. The concentration of lactic acid (LD) and the specific activities of LDH in liver and skeletal muscle of two species were measured. The different isoenzymes of LDH were determined by native polyacrylamide gel electrophoresis (PAGE). The results showed that, (1) LDH-B mRNA level in skeletal muscle of plateau zokor was significantly higher than that of plateau pika (P < 0.01), but no differences was found at LDH-A mRNA levels between them (P > 0.05); (2) PC, LDH-A and LDH-B mRNA levels in liver of plateau pika were significantly higher than those of plateau zokor (P < 0.01); (3) The LDH activity and concentration of LD in skeletal muscle and liver, as well as the PC activity in liver of plateau pika were significantly higher than those of plateau zokor (P < 0.01); (4) The isoenzymatic spectrum of lactate dehydrogenase showed that the main LDH isoenzymes were LDH-A4, LDH-A3B and LDH-A2B2 in skeletal muscle of plateau pika, while the main LDH isoenzymes were LDH-AB3 and LDH-B4 in skeletal muscle of plateau zokor; the main isoenzymes were LDH-A3B, LDH-A2B2, LDH-AB3 and LDH-B4 in liver of plateau pika, while LDH-A4 was the only isoenzyme in liver of plateau zokor. These results indicate that the plateau pika gets most of its energy for sprint running through enhancing anaerobic glycolysis, producing more lactate in their skeletal muscle, and converting lactate into glucose and glycogen in the liver by enhancing gluconeogenesis. As a result, the plateau pika has a reduced dependence on oxygen in its hypoxic environment. In contrast, plateau zokor derives most of its energy used for digging activity by enhancing aerobic oxidation in their skeletal muscle, although they inhabit hypoxic underground burrows.
Animals ; Glycolysis ; Hypoxia ; metabolism ; Isoenzymes ; metabolism ; L-Lactate Dehydrogenase ; metabolism ; Lactic Acid ; metabolism ; Lagomorpha ; physiology ; Liver ; enzymology ; Muscle, Skeletal ; enzymology ; Oxygen ; metabolism ; RNA, Messenger

BACKGROUNDSeveral studies suggest that cyclooxygenase-2 (COX-2) contributes to the delayed progression of ischemic brain damage. This study was designed to investigate whether COX-2 inhibition with parecoxib reduces focal cerebral ischemia/reperfusion injury in rats.

METHODSNinety male Sprague-Dawley rats were randomly assigned to three groups: the sham group, ischemia/reperfusion (I/R) group and parecoxib group. The parecoxib group received 4 mg/kg of parecoxib intravenously via the vena dorsalis penis 15 minutes before ischemia and again at 12 hours after ischemia. The neurological deficit scores (NDSs) were evaluated at 24 and 72 hours after reperfusion. The rats then were euthanized. Brains were removed and processed for hematoxylin and eosin staining, Nissl staining, and measurements of high mobility group Box 1 protein (HMGB1) and tumor necrosis factor-α (TNF-α) levels. Infarct volume was assessed with 2,3,5-triphenyltetrazolium chloride (TTC) staining.

RESULTSThe rats in the I/R group had lower NDSs (P < 0.05), larger infarct volume (P < 0.05), lower HMGB1 levels (P < 0.05), and higher TNF-α levels (P < 0.05) compared with those in the sham group. Parecoxib administration significantly improved NDSs, reduced infarct volume, and decreased HMGB1 and TNF-α levels (P < 0.05).

CONCLUSIONSPretreatment with intravenous parecoxib was neuroprotective. Its effects may be associated with the attenuation of inflammatory reaction and the inhibition of inflammatory mediators.

Animals ; Blotting, Western ; Brain Ischemia ; drug therapy ; metabolism ; prevention & control ; Injections, Intravenous ; Isoxazoles ; administration & dosage ; therapeutic use ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; drug therapy ; metabolism ; prevention & control

OBJECTIVETo study the expression of mPGES-1 in hepatocellular carcinoma (HCC), observe the effect of MK886 on down-regulation of mPGES-1 gene expression on the biology of human hepatocarcinoma cell line HepG2 and to investigate its significance in the occurrence, progression, metastasis and invasion.

METHODSHCC tissues, para-carcinoma tissues, far-carcinoma tissues and normal liver tissues were collected. The expressions of mPGES-1 were determined by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. The proliferation, adherence, migration and invasion abilities of HepG2 cells interfered by MK886 were assessed by MTT and transwell technique respectively.

RESULTSThe expression of mPGES-1 in HCCs was higher than that in normal liver tissues (P < 0.01), which increased following histological grade. Furthermore, mPGES-1 expression level was higher in the capsule invasion and metastasis tumor than in primary locus. A significant dose-dependent down-regulation of expressions of mPGES-1 gene mRNA and protein were observed in HepG2 cells when MK886 was given for 48 h (F = 140.402, P < 0.01; a'= 0.00714, P < 0.01). Compared with the control group, the growth inhibitory rate of HepG2 cell was observed significantly time and dose-dependent when MK886 was given. The rate of adhesion cells in experimental groups were 85.3% ± 1.3%, 70.5% ± 1.5% and 45.8% ± 2.4%, respectively, less than that in control group 100.0% ± 0 (F = 626.313, P < 0.01). The migration cells was 92.47 ± 1.90, 62.63 ± 1.96 and 37.33 ± 0.83 respectively in the experimental groups after 24 h, lower than that in the control group 128.93 ± 2.60 (F = 1253.805, P < 0.01). The invasion assay revealed that the invading cells were 41.67 ± 1.30, 25.47 ± 1.30 and 13.93 ± 1.66 in the experimental groups, in contrast to 55.67 ± 2.08 in control group after 24 h. The difference between these groups was significant (F = 372.615, P < 0.01). The numbers of adhesion, migration and invasion of HepG2 cells were dose-dependent in MK886 groups.

CONCLUSIONOver-expression of mPGES-1 was associated with the tumorigenesis and progression of HCC. The down-regulation of mPGES-1 gene expression might indicated the decrease of the invasion and metastasis of HCC.

Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Adhesion ; Cell Movement ; Cell Proliferation ; Female ; Hep G2 Cells ; Humans ; Indoles ; pharmacology ; Intramolecular Oxidoreductases ; metabolism ; Liver Neoplasms ; metabolism ; pathology ; Male ; Microsomes ; metabolism ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Prostaglandin-E Synthases

OBJECTIVETo investigate the expression of protein kinase C (PKC) in the spinal dorsal horn of rats with formalin-induced pain and the effect of intrathecal ketamine on PKC expression.

METHODSThirty-two SD rats were randomly divided into 4 equal groups, namely the control group, intrathecal saline group (NS), 50 µg ketamine group (K1) and 100 µg ketamine group (K2). The rats were anesthetized with 10% chloral hydrate, and a microspinal catheter was inserted intrathecally into the lumbar region. Five days later, the rats in groups, K1 and K2 were subjected to intrathecal administration of 50 and 100 µg ketamine (10 µl), respectively, followed by 10 µl saline, and those in NS group received 20 µl saline only. Thirty minutes later, 5% formalin (50 µl) was subcutaneously injected into the left hindpaw. The pain intensity score (PIS) was utilized to assess antinociceptive behavior within 1 h after formalin injection. Twenty-four hours later, the left hindpaw thickness was measured and the expression of PKC in the spinal dorsal horn in the L5 segment was assayed using immunohistochemistry.

RESULTSCompared to group NS, groups K1 and K2 showed significantly decreased PIS (P<0.01) in the second phase of formalin-induced pain; 24 h later, the left hindpaw thickness of group NS increased obviously in comparison with that in the control group (P<0.01), whereas the thickness was significantly reduced in group K1 and K2 as compared to that in group NS (P<0.05). The number of immunoreactive cells and the immunohistochemical score of PKC in the spinal dorsal horn were significantly higher in group NS than in group C (P<0.01), but significantly lower in groups K1 and K2 than in group NS (P<0.05).

CONCLUSIONIntrathecal ketamine produces obvious antinociception against formalin-induced pain in rats and inhibits the enhanced PKC expression in the spinal dorsal horn in response to formalin-induced pain, suggesting the important role of PKC in nociceptive signal transmission and modulation in the spinal cord.

Animals ; Formaldehyde ; adverse effects ; Injections, Spinal ; Ketamine ; administration & dosage ; pharmacology ; Male ; Pain ; chemically induced ; metabolism ; Pain Measurement ; Posterior Horn Cells ; metabolism ; Protein Kinase C ; metabolism ; Rats ; Rats, Sprague-Dawley ; Spinal Cord ; drug effects ; metabolism

OBJECTIVE: To explore the clinical efficacy of intrathecally administered low dose sufentanil-bupivacaine in transurethral resection of the prostate (TURP). METHODS. Ninety patient (ASA I - III) undergoing TURP were randomly divided into 3 groups (n = 30); Group A, B and C. Group A received 7.5 mg bupivacaine + 5 microg sufentanil + 10% glucose; Group B received 7.5 mg bupivacaine + 7.5 microg sufentanil + 10% glucose; Group C received 15 mg bupivacaine + 10% glucose. The volume was 3 mL in every group. SP, DP, HR, SpO2, the degree of motor and sensory blockade and the side effect were observed. RESULTS: SP/DP was significantly decreased in Group C than that in Group and Group B (p<0.05), HR and SpO2 in group B were decreased to different degrees 15 min after the injection (p<0.05). The complete recovery time of motor nerve blockade and the regression time of sensory blockade were obviously prolonged in Group C (p<0.05). There were no significant differences in analgesic effect among the three groups during the operation, but the incidence of pruritus was higher in both group A and Group B than that in Group C during the first 24 hours after the injection. CONCLUSION Spinal anesthesia with low dose sufentanil-bupivacaine possesses relatively steady hemodynamics. The blockade degree of motor and sensory blockade in this spinal anesthesia is lower than that in standard spinal bupivacaine in TURP.
Aged ; Anesthesia, Spinal ; Bupivacaine ; administration & dosage ; Humans ; Male ; Middle Aged ; Sufentanil ; administration & dosage ; Transurethral Resection of Prostate

OBJECTIVE: To evaluate the immunological function in rats with formalin inflammatory pain through intrathecal pumping different dosages of morphine. METHODS: Thirty-two Sprague-Dawley rats were randomly divided into 4 groups (n = 8 in each group): saline group (NS) and morphine group included M1 group (10 microg/h) , M2 group (5 microg/h), and M3 group (2.5 microg/h). Chronic intrathecal catheterization was performed under anesthesia with 10% chloral hydrate (300-350) mg/kg according to M2 group (5 microg/h) and M3 group (2. 5 microg/h). Chronic intrathecal catheterization was modified Yaksh's. After 7 days, pain intensity scoring (PIS) was utilized to assess antinociceptive effect of morphine. And spleens were aseptically removed to obtain splenic cells. T lymphocyte function was evaluated based on Concanavalin-A induced splenocyte proliferation. A modified lactic acid dehydrogenase release assay was used to assess NK cell activity. Phenotypic expression of cell surface markers of T lymphocyte subsets (CD3+, CD3+ CD4+, CD3+ CD8+, and CD4+ / CD8+ ) and NK cell ( CD161+) in the spleen were analyzed by flow cytometry. RESULTS: Compared with the NS group, PIS of morphine group decreased obviously (P < 0.01) and was dose-dependent in the early and late phase of formalin pain, but there were no significant differences among morphine groups. Spleen index, splenocyte proliferation and NK cell activity were significantly suppressed by intrathecal pumping morphine. Phenotypic expression of T lymphocyte subsets and NK cell assessed by flow cytometry were different from the control group in all morphine groups. CONCLUSION There was significant antinociception of intrathecal pumping morphine. After intrathecal pumping different dosages of morphine (10 microg/h,5 microg/h, and 2.5 microg/h), the function of cellular immunity was suppressed and was dose-dependent.
Analgesics, Opioid ; administration & dosage ; pharmacology ; Animals ; Dose-Response Relationship, Drug ; Formaldehyde ; Immunosuppressive Agents ; administration & dosage ; pharmacology ; Injections, Spinal ; Killer Cells, Natural ; immunology ; Male ; Morphine ; administration & dosage ; pharmacology ; Pain ; chemically induced ; immunology ; Pain Measurement ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; T-Lymphocyte Subsets ; immunology

OBJECTIVE: To evaluate the effect of intrathecal pumping tramadol on cell-mediated immunity in rats with formalin inflammatory pain. METHODS: Thirty-two Sprague-Dawley adult male rats weighting 250 approximately 300 g were randomly divided into 4 groups (n=8 in each group):Saline group (NS) and 3 tramadol groups (T1,T2,and T3). The rats were anesthetized with intraperitoneal chloral hydrate (300 approximately 350)mg/kg. Microspinal catheter was inserted into the subarachnoid space at the lumber region according to modified Yaksh techniques. In the tramadol groups,after 5 days tramadol was continuously infused through the spinal catheter at 50 (T1),25 (T2), and 12.5 microg/h (T3) for 7 days. In the NS group normal saline was continuously infused instead of tramadol. On Day 7 formalin (5%, 50 microL) was injected into the plantar surface of the left hindpaw. The number of flinches, lickings and total time of licking was recorded for 60 min.Pain intensity scoring(PIS)(0 approximately 3;0= no pain, 3=severe pain) was used to assess the antinociceptive effect of intrathecal tramadol. The rats were killed after the evaluation of pain intensity. Body weight and spleen weight were measured and spleen index (spleen weight/body weight) was calculated. T-lymphocyte function was evaluated based on Concanavalin-A(ConA) induced splenocyte proliferation. A modified lactic acid dehydrogenase(LDH) release assay was done to assess the NK cell activity. Phenotypic expressions of cell surface markers of T lymphocyte subsets (CD3+, CD3+ CD4+, CD3+ CD8+, and CD4+/ CD8+) and NK cell(CD161+) in the spleen were analyzed by flow cytometry. RESULTS: The PIS scores were significantly lower in the T1,T2,and T3 groups than those in the NS group. The spleen index and splenocyte proliferation induced by ConA were significantly suppressed in the T1 group,and the phenotypes of T lymphocyte subsets were significantly changed,but no significant difference was found in the T2 and T3 groups compared with the NS group. There were no differences in NK cell activity in the 3 tramadol groups from the control group. CONCLUSION Intrathecal pumping tramadol has significantly antinociceptive effect. Intrathecal pumping higher dosage tramadol (50microg/h) suppresses T lymphocyte proliferation and alteres T lymphocyte subset phenotype but does not affect NK cell activity. General analgesic dosage tramadol (25 and 12.5 microg/h) has no effect on the immune function.
Analgesics, Opioid ; pharmacology ; Animals ; Dose-Response Relationship, Drug ; Formaldehyde ; Injections, Spinal ; Killer Cells, Natural ; immunology ; Male ; Pain ; chemically induced ; immunology ; Pain Measurement ; drug effects ; Rats ; Rats, Sprague-Dawley ; T-Lymphocyte Subsets ; immunology ; Tramadol ; administration & dosage ; pharmacology