AIMTo explore the effects of intrathecal injection of selective cyclooxygenase-1 (COX-1) inhibitor, SC-560, on mechanical allo dynia and spinal ERK protein expression in rats with postoperative pain.

METHODSRats were divided into 4 groups: control group, postoperative pain group, SC-560 group and DMSO group. Mechanical withdrawal threshold (MWT), immunohistochemical and Western blotting technique were used to evaluate mechanical hypersensitivity and the expression of phospho-ERK in the spinal cord, respectively.

RESULTS(1) Behavior test rats developed allodynia 1 h after operation and SC-560 100 microg administrated intrathecally demonstrated a significant reduction in postoperative hypersensitivity. (2) Immunohistochemical staining Phospho-ERK positive neurons in the rat superficial spinal dorsal horn increased significantly 1 h after incision compared with that of non-incision group. Intrathecal administration of SC-560 preoperatively could significantly reduce the number of phospho-ERK positive neurons. (3) Western blot expression of phospho-ERK1/2 protein in the lumbar spinal cord increased significantly 1 h after incision and decreased by intrathecal injection of SC-560.

CONCLUSIONSC-560 administrated intrathecally can inhibit mechanical hypersensitivity induced by postoperative pain in rats and this anti-allodynic process may mediated by spinal ERK.

Animals ; Cyclooxygenase Inhibitors ; pharmacology ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Male ; Pain Measurement ; drug effects ; Pain, Postoperative ; metabolism ; Pyrazoles ; pharmacology ; Rats ; Rats, Wistar ; Spinal Cord ; drug effects ; metabolism

OBJECTIVETo study the regulation of luteolin on spleen cells and sarcoma S180 cells in normal ICR mice.

METHODSSpleen cells and S180 cells were incubated with different concentrations of luteolin (50, 100, 200, and 400 μmol/L). The effect of luteolin on spleen cells and sarcoma S180 cells was determined by MTT assay. The apoptosis was detected using propidium iodide staining flow cytometry. Intracellular reactive oxygen species (ROS) was determined by flow cytometric analysis. Activities of free radicals scavenging were determined by hydroxyl radical and DPPH tests.

RESULTSCompared with the solvent control group, 200 and 400 μmol/L luteolin increased the spleen cells viability (P < 0.05). Luteolin at 100, 200, and 400 μmol/L decreased activities of S180 cells (P < 0.01). The proportion of sub-G1 phase spleen cells was reduced after treated with 200 and 400 μmol/L luteolin (P < 0.05). The proportion of sub-G1 phase S180 cells was elevated after treated with 200 and 400 μmol/L luteolin (P < 0.05). Compared with the solvent control group, levels of intracellular ROS in spleen cells of ICR mice all increased; levels of intracellular ROS in S180 cells all decreased after treated with 50, 100, 200, and 400 μmol/L luteolin (P < 0.05). Luteolin scavenged hydroxyl radical and DPPH in a dose dependent manner.

CONCLUSIONLuteolin had bilateral regulation on viability and apoptosis of spleen cells and S180 cells (promoting the viability of spleen cells, inhibiting apoptosis of spleen cells, inhibiting the viability of S180 cells, and promoting apoptosis of S180 cells), which was worth further study and exploration.

Animals ; Apoptosis ; Apoptosis Regulatory Proteins ; metabolism ; Cell Survival ; Luteolin ; metabolism ; Mice ; Mice, Inbred ICR ; Reactive Oxygen Species ; Sarcoma ; Spleen ; metabolism

KRAS is one of the most frequently mutated human oncogenes. In spite of mounting efforts on the development of direct or indirect inhibition targeting KRAS, little has been achieved because of insurmountable difficulties, titling KRAS "undruggable". Recently, subtype-specific inhibitors have shown great hope. Some KRAS^G12C inhibitors have entered clinical trials, including adagrasib and sotorasib, and have shown preliminary clinical effectiveness. Experiences from the inhibitors targeting the downstream factors of RAS pathways show that the anticancer activity of these drugs will be limited due to the development of drug resistance. Preclinical studies of KRAS^G12C inhibitors have revealed that the application of these agents might be hampered by the drug resistance issue. The current review aims to describe the current status of KRAS^G12C inhibitors, and discuss the mechanisms underlying KRAS^G12C inhibitor resistance, so as to provide the clues for the combat of drug resistance.

OBJECTIVETo investigate the effects of ropivacaine on Gamma-aminobutyric acid(GABA)-activated currents in dorsal root ganglion (DRG) neurons in rats and discuss the analgesia mechanism of ropivacaine.

METHODSBy means of using whole-cell patch-clamp technique, to investigate the modulatory effects of ropivacaine on GABA-activated currents (I(GABA)) in acutely isolated dorsal root ganglion neurons.

RESULTS(1) In 48 out of 73DRG cells (65.7%, 48/73), to perfusion ropivacaine bromide (0.1 - 1 000 micromol/L) were sensitive. Which produce in 0 to 380 pA current. (2) The majority of the neurons examined (74.5%, 73/98) were sensitive to GABA. Concentration of 1 - 1 000 micromol/L GABA could activate a concentration-dependent inward current, which manifested obvious desensitization, and the inward currents could be blocked byGABA-receptor selective antagonist of bicuculline (100 micromol/L). (3) After the neurons were treated with ropivacaine (0.1 - 1000 micromol/L) prior to the application of GABA (100 micromol/L) 30 s, GABA currents were obviously increased. Ropivacaine could make dose-response curve of the GABA up, EC50 is 23.46 micromol/L. Ropivacaine shifted the GABA dose-response curve upward and increased the maximum response to the contrast about 153%.

CONCLUSIONThe enhancement of ropivacaine to DRG neurons activation of GABA current, can lead to enhancement of pre-synaptic inhibition at the spinal cord level. This may be one of the reasons for the anesthetic effect and analgesia for ropivacaine in epidural anesthesia.

Amides ; pharmacology ; Animals ; Ganglia, Spinal ; cytology ; physiology ; Membrane Potentials ; drug effects ; Neurons ; cytology ; drug effects ; physiology ; Patch-Clamp Techniques ; Rats ; Rats, Sprague-Dawley ; Receptors, GABA-A ; physiology

Objective To study the effect of Sinisan on the ultra structure of hippo-campus to the intervention in rats with post-traumatic stress disorder(PTSD).Methods SD rats were divided equally into 5 groups,each group had ten rats:blank control group,model group,negative control group,positive control group and experimental group.The blank control group did not copy the model,the normal feeding.Model group,repetitive post-traumatic stress disorder model was induced by current stimulation,but not treated.Negative control group,equal volume of 0.9％ NaC1.Positive control group (paroxetine hydrochloride 0.42 mg · mL-1) and experimentalgroup (Sinisan,containing crude drug 0.24 g · mL-1).It given to drugs 1 h before the model establishment.The rats were administered with 10 mL · kg-1,once a day,for a total of 7 d.In each group,rats was cardiac perfusion and the hippo-campus tissue was collected.The changes of ultra structure of CA1 and CA3 area in hippo-campus with rats were observed by transmission electron microscope.Results In the blank control group,the CA1 and CA3 neurons in the hippocampus were rich in cytoplasmic organelles,and the mitochondria were round or long,the structure of the mitochondrial cristae was clear,the rough endoplasmic reticulum was like a cord like distribution,and the ribosome was abundant.Compared with the blank control group,the organelle of CA1 and CA3 area in hippo-campus of the model group were significantly damaged,the cytoplasm was open,the mitochondria were swollen,the mitochondrial membrane and cristae disappeared,and the rough endoplasmic reticulum was dilated.The results showed that the damage of the organelles in the hippocampal neurons of rats was induced by the electric shock.The model group was similar to the negative control group,which indicated that there was no significant effect on the stress of rats after intragastric administration.The intracellular organelles in the CA1 and CA3 neurons in the hippocampus of the positive control group and the experimental group were significantly recovered,and the changes in the two groups were similar.These results suggest that both paroxetine hydrochloride and Sinisan can significantly improve the structure of CA1 and CA3 neurons in the hippocampus of PTSD rats.Conclusion Snisan as traditional Chinese medicine compound can significantly improve the hippocampus of rats with PTSD CA1 and CA3 neurons.

Objective To study on intervene function of Sinisan with the ultra structure of hippo-campus in rats with sleep disorder induced by post traumatic stress disorder.Methods Fifty SD rats were divided equally into five groups:blank control group,model group,negative control group,positive control group and experimental group.The control blank group did not copy the model,do not receive drugs,the normal feeding.Model group,repetitive post-traumatic stress disorder model was induced by current stimulation,but not receive drugs.Negative control group,equal volume of 0.9％ NaCl.Positive control group (paroxetine hydrochloride 0.42 mg ·mL-1) and experimental group (Sinisan,containing crude drug 0.24 g · mL-1).It given to drugs 1 h before the model establishment.The rats were administered with 10 mL · kg-1,once a day,for a total of 7 d.The changes of ultra structure of CA1 and CA3 area in hippo-campus with rats were observed by transmission electronmicroscope.Results In the blank control group,the hippocampal neurons were clearly defined,and the synaptic space was obvious in the CA1 and CA3 area.Compared with the blank control group,in the model group,the synaptic structure of hippocampal neurons was significantly changed,the synaptic vesicles were reduced and the synaptic structure was not clear in the CA1 and CA3 area.Compared with the model group,the negative control group was same too,which indicated that there was no significant stress effect on the rats after intragastric administration of saline.Compared with the negative control group,positive control group and experimental group hippocampal neurons synapse structure was restored,and the two groups were similar in the CA1 and CA3 area.Conclusion Transmission electronmicroscope was used to study the ultra structure of CA1 and CA3 in rats hippo-campus with characteristic difference.

Objective To study the effects of sleep latency to the intervention of Sinisan in rats with post-traumatic stress and sleep disorder (PTSD).Methods Fifty SD rats were divided equally into 5 groups:sham operation group,model group,negative control group group,positive control group and experimental group.PTSD model was made by claustrophobia,but not in sham operation group.The model group was not given the drug,the negative control group was given equal volume 0.9％ NaC1,and the positive control group was given paroxetine hydrochloride 0.42 mg · mL-1.The experimental group was perfused with the decoction of Sinisan (containing 0.24 g · mL-1) 10 mL · kg-1.The drug was administered 1 h before the stress model was administered once a day for a total of 7 d.After intervention on the 7th day,nonrapid eye movements sleep (NREMS) and eye movements sleep (REMS) were detected.Results The REMS and NREMS of sham operation group and model group were respectively (8.66 ± 3.04),(23.27 t 10.15) min;(65.90 ± 25.08),(109.36 ± 43.43) min,the differences between groups were statistically significant(all P ＜ 0.01);the result suggest that difficulty in falling asleep appears in rats after modeling.The REMS and NREMS of the positive control group and the experimental group were respectively (8.17 ±2.29),(6.83 ±2.84) min;(162.29 ±46.19),(195.24 ±67.96) min.Comparison between the drugs groups and model group,the difference was statistically significant (P ＜0.05,P ＜0.01).These Results suggested that both paroxetine hydrochloride and Sinisan can significantly promote the sleep of rats.Conclusion Traditional Chinese medicine compound Sinisan can obviously promote the rats with sleep disorder caused by PTSD.

Objective To investigate the possible mechanism of lycopene on protecting against hypoxia/reoxygenation (H/R)-injury.Methods Primary cultured cardiomyocytes,isolated from neonatal mouse,were divided into three groups randomly:control group(C) ; H/R group(4 h H followed by 8 h R) ;lycopene + H/R group(L + H/R),in which the cardiomyocytes were pretreated with lycopene for 4 h before H/R.The survival of cardiomyocytes was counted.Apoptotic cells were detected by TUNEL assays.The release of cytochrome c from mitochondrial matrix into the cytosol,the activity of caspase-3,intracellular ROS levels and the activity of calpain were also determined in these groups respectively at the same time.Results The pretreatment of cardiomyocytes with lycopene significantly improved the survival of cardiomyocytes [C:(89.84 ± 5.15) ％,H/R:(63.59 ± 5.11) ％,L + H/R:(79.25 ± 1.48) ％,P ＜0.05] and reduced the extent of apoptosis [C:(10.37 ± 1.25) ％,H/R:(32.03 ± 4.79) ％,L + H/R:(22.57 ± 3.22) ％,P ＜ 0.05],significantly reduced caspase-3 activation [C:(2.61 ± 0.19),H/R:(5.82±0.92),L + H/R:(3.74 ±0.64) pNA pmol/μg protein,P ＜0.05].To further study the mechanism underlying the benefits of lycopene,interactions between lycopene and calpain activation were examined.Lycopene pretreatment of cardiomyocytes suppressed the activation of calpain (C:272.33 ±300.46,H/R:1156.00 ± 212.02,L + H/R:607.33 ± 166.23,P ＜ 0.05) by reducing the H/R induced increased intracellular ROS levels [C:100％,H/R:(239.79 ± 27.27)％,L + H/R:(188.19 ±17.63) ％,P ＜ 0.05].Conclusion Lycopene may protect against hypoxia/reoxygenation-induced injury by preventing calpain activation.

OBJECTIVETo observe the effect of crystalline NiS on genome DNA methylation profile in in vitro cultured cells.

METHODS16HBE Cells were treated with crystalline NiS at 0.25, 0.50, 1.00 and 2.00 µg/cm(2) for 24 h and three times at total. DAC treatment was given at 3 µmol/L for 72 h.5-mC immunofluorescence and SssI methyltransferase assay methods were applied to investigate if the hypomethylation of genome DNA involved.

RESULTSThe results of 5-mC immunofluorescence showed that the fluorescence intensity of NiS-treated cells were decreased in some degree, and transformed cells were decreased dramatically. By the SssI methylase assay, an average of (81.9 ± 7.3)% methylated CpG were found in negative control cells. By contrast, (77.9 ± 6.2)%, (75.3 ± 6.8)%, (59.5 ± 4.9)%, (67.4 ± 5.1)% methylated CpG were observed in cells treated with NiS for three times at dosage of 0.25, 0.50, 1.00 and 2.00 µg/cm(2) which were abbreviated as NiS0.25, NiS0.50, NiS1.00, NiS2.00 respectively. The ANOVA analysis results showed that there was a significant difference in the 5 groups above (F = 124.95, P < 0.01). The results of Dunnett-t test showed that the methylated CpG of both group NiS1.00 and NiS2.00 were significantly decreased compared with the negative control group (t values were 7.64, 4.89 respectively, P < 0.01). For methylated CpG, (46.2 ± 4.1)% and (43.6% ± 4.3)% were observed in NiS-transformed cells (NSTC1 and NSTC2) which were dramatically decreased compared with the negative control group (t values were 12.79, 13.56 respectively, P < 0.01).

CONCLUSIONGenomic DNA methylation levels were decreased during NiS induced malignant transformation.

Bronchi ; cytology ; Cell Line ; Cell Transformation, Neoplastic ; chemically induced ; DNA Methylation ; drug effects ; Epithelial Cells ; drug effects ; Genome ; Humans ; Nickel ; adverse effects ; chemistry

OBJECTIVETo determine occupational hazards in work sites of a large solid waste landfill and analyze their adverse health effects.

METHODThe national standardized detection methods were used to determine dust concentration, harmful gas and physical factors in worksites. Routine physical examination, pulmonary function, hearing tests and nervous system test were performed in workers for 2 consecutive years. Urine lead, cadmium and mercury contents were detected. The comet assay was use to measure DNA damage in peripheral blood lymphocytes among workers.

RESULTThe main occupational hazard factors in this solid landfill are dust, harmful gas, high temperature and noise. The oxides, carbon monoxide, and noise and high temperatures in summer at some work sites exceeded the national occupational exposure limits. The prevalence of respiratory inflammation and rate of pulmonary function decrease among front-line workers and on-site technical managers are 21.2% and 11.5%, which are significantly higher than those among administrative staff (7.1% and 0) (P < 0.05). Nervous system abnormalities rate of front-line workers and on-site technical managers was 50.0%, which is significantly higher than that (26.7%) of administrative staff (P < 0.05). Because of long-term exposure to high intensity noice, hearing loss rate of bulldozer drivers was 10.3%. In addition, about 75% of workers with DNA damage in peripheral blood lymphocyte are front-line workers.

CONCLUSIONAdverse health effects from occupational hazards were observed among workers in this solid waste landfill.

Adult ; Female ; Humans ; Male ; Middle Aged ; Occupational Diseases ; epidemiology ; Occupational Exposure ; Refuse Disposal ; Respiratory Tract Infections ; epidemiology ; Risk Factors ; Workplace