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 possible mechanism of lycopene on protecting against hypoxia/reoxygenation (H/R)-injury.

METHODSPrimary 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.

RESULTSThe 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].

CONCLUSIONLycopene may protect against hypoxia/reoxygenation-induced injury by preventing calpain activation.

Animals ; Apoptosis ; drug effects ; Calpain ; metabolism ; Carotenoids ; pharmacology ; Caspase 3 ; metabolism ; Cell Hypoxia ; Cells, Cultured ; Cytochromes c ; metabolism ; Mice ; Mice, Inbred C57BL ; Myocytes, Cardiac ; drug effects ; metabolism ; pathology

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

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

OBJECTIVEThis study was to explore the differences in the nucleoprotein gene between rabies virus (RABV) and its vaccine strains in Guizhou province from year 2005 to 2010.

METHODSSamples from 4 rabies patients and cerebral tissue samples of 28 rabies infected dogs were collected from different districts in Guizhou province between year 2005 and 2010. Direct Immunofluorescence Assay (DFA) and RT-nested PCR assay were applied to detect the overall length of N gene sequence. Meanwhile, based on the comparison between the homology and phylogenetic tree, the differences in N gene sequence between the prevalent RABV and the RABV vaccine strains collected from NCBI database in these years.

RESULTSAccording to DFA and RT-nested PCR assay, the antigen and nucleic acid of the 21 dogs and 4 human samples were both confirmed positive; whose full length of N gene sequences were both 1353 bp. The homological analysis showed that the 25 strains of RABV virus and the RABV type I virus stored by GenBank database shared a high homology in N gene nucleotide and amino acid sequences, which were 89%-100% and 98%-100%, respectively. Besides, the homology between the 25 strains of RABV virus and its vaccines in nucleotide and amino acid sequences were separately 86%-95% and 96%-100%. The N gene of vaccines for livestock shared the highest homology with HEP-Flury strain in the nucleotide and amino acid, which were 88%-89% and 98%-99%, respectively. The vaccines for human use showed its greatest homology with the CTN strain in nucleotide (86%-100%) and amino acid (96%-100%). The phylogenetic tree analysis indicated that the 25 strains of RABV virus, RABV type I virus and the CTN vaccine strains constituted one individual cluster, which was least different from the CTN vaccine for human use.

CONCLUSIONThe prevalent RABV virus, the vaccine HEP-Flury for livestock and the vaccine CTN for human use were found to be highly similar in N gene expression in Guizhou province from 2005 to 2010.

Amino Acid Sequence ; Animals ; Dogs ; Genotype ; Humans ; Molecular Sequence Data ; Nucleoproteins ; genetics ; RNA, Viral ; genetics ; Rabies ; veterinary ; virology ; Rabies Vaccines ; genetics ; Rabies virus ; classification ; genetics ; isolation & purification

OBJECTIVETo compare the efficacy between subtotal colectomy with antiperistaltic cecoproctostomy and total colectomy with ileorectal anastomosis (TAC-IRA) for patients with severe refractory slow transit constipation(STC).

METHODSDuring 1999 to 2002, TAC-IRA was the preferred procedure for 20 STC patients in our department. From 2003 to 2005, 17 STC patients underwent subtotal colectomy plus antiperistaltic cecoproctostomy. Clinical data of the two groups were collected and compared retrospectively.

RESULTSThere were no significant differences in basic preoperative clinical data between the two groups. During the follow-up period, the time of daily defecation in the antiperistaltic cecoproctostomy group was less than that of TAC-IRA group (2.4+/-0.9 vs 3.4+/-0.8, P=0.0014), meanwhile the Wexner continence score was significantly lower in the antiperistaltic cecoproctostomy group (4.3+/-1.8 vs 5.8+/-1.9, P=0.0223). Barium enema after subtotal colectomy showed that residual ascending colon and cecum presented a sign of "reservoir".

CONCLUSIONSubtotal colectomy with antiperistaltic cecoproctostomy is a better method for appropriately selected patients with STC than TAC-IRA.

Adult ; Aged ; Anastomosis, Surgical ; Cecum ; surgery ; Colectomy ; methods ; Colon ; surgery ; Constipation ; surgery ; Female ; Humans ; Male ; Middle Aged ; Rectum ; surgery ; Retrospective Studies

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

BACKGROUNDTransplantation of adult bone marrow-derived mesenchymal stem cells (MSCs) has been proposed as a strategy for cardiac repair following myocardial damage. However cell transplantation strategies to replace lost myocardium are limited by the inability to deliver large numbers of cells that resist peritransplantation graft cell death. Accordingly, we set out to isolate and expand adult swine bone marrow-derived MSCs, and to engineer these cells to overexpress AKT1 (protein kinase B), to test the hypothesis that AKT1-engineered MSCs are more resistant to apoptosis and can enhance cardiac repair after transplantation into the ischemic swine heart.

METHODSThe CDS (regulation domain of AKT1) AKT1-cDNA fragment was amplified, and MSCs were transfected following synthesis with a pCDH1-AKT1 shuttling plasmid. Western blotting analysis and real-time reverse transcription-polymerase chain reaction (RT-PCR) was performed. Myocardial infarction (MI) models were constructed in Meishan pigs, and cardiac function was evaluated by magnetic resonance imaging (MRI) measurements and echocardiography 4 weeks later. All pigs were assigned to four groups: control (A), DMEM (B), MSC (C), and AKT-transfected (D). MSCs were transfected with the AKT1 gene, and autologous BrdU-labeled stem cells (1 x 10(7)/5 ml) were injected into left anterior descending coronary atery (LAD) of the infarct heart in groups C and D. In group B, DMEM was injected using the same approach. In group A, there was no injection following LAD occlusion. After 4 weeks, cardiac function and regional perfusion measurements were repeated by MRI and echocardiography, and histological characteristics of the hearts were assessed. Connecxin-43 (CX-43), BrdU, and von Willebrand factor (VWF) immunoreactivity was tested using enzyme linked immunosorbent assay (ELISA). Vascular endothelial growth factor (VEGF), transforming growth factor-beta1 (TGF-beta1) were analyzed at the same time.

RESULTSAKT1-cDNA was cloned into pCDH1-MCS1-EF1-copGFP and the sequence was confirmed. AKT mRNA expression was detected at 24 hours after transfection. AKT1 expression in MSCs remained strong after 2 weeks, according to real-time RT-PCR and Western blotting. Prior to cell implantation, end-diastolic left ventricular dimension (EDLVd) increased and stroke volume (SV) decreased in the MI hearts. MRI scans revealed significantly improved cardiac function following implantation, and implanted MSCs prevented thinning and expanding in the infarct region, as well as improved contraction and increased perfusion in all groups compared to control hearts. The left ventricular chamber size was smaller in cell-transplanted hearts than in control hearts. Moreover, group D exhibited significant improvement. The expression of CX-43, BrdU, and VWF could be found in the immunohistochemical pathological sections of group C and group D. The level of VEGF reached a high level 1 week after implanting the MSCs, but the level of TGF-beta1 decreased gradually.

CONCLUSIONSThe AKT1-expressing lentiviral vector resulted in stable over-expression of AKT1 in MSCs. MSC engraftment in host myocardium improved cardiac function by attenuating contractile dysfunction and pathological thinning of the infracted left ventricular wall, which likely resulted from myocardial regeneration and angiogenesis.

Animals ; Blotting, Western ; Disease Models, Animal ; Electrophoresis ; Genetic Vectors ; genetics ; Lentivirus ; genetics ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; metabolism ; Myocardial Infarction ; metabolism ; therapy ; Proto-Oncogene Proteins c-akt ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Swine ; Ventricular Remodeling ; physiology