Rats were immersed into boiling water for 15 seconds with scalded area about 23 cm~2 and decapitated at 0.5,1 and 6 hours after scalding. The changes of immunoreactive glueocorticoid receptor (GR_(IR)) and glucocorticoid receptor binding (GR_B) of hepatic cytosol after scalding were studied by both monoclonal antibody method and radioligand binding assay. The results showed that GR_(IR) and GR_B decreased markedly in parallel manner after scalding. Comparing with the controls (100%), the values of GRIR of 0.5,1 and 6 hours after scalding decreased to 64.76?13.320% (n=4),59.55?13.18% (n=5) and 42.64?4.24% (n=4)respectively; and relevent GR_B decreased to 50.24?9.71% (n=4), 51.11?11.31% (n=5) and 30.43?3.03% (n=4). The amplitude of decrease of GR_B was about 10% more than that of GR_(IR). Thus it might be concluded that the reduction of GR_B after stress was mainly due to the decrease of GR protein molecules but 10% of that was due to the change of GR binding activity. The mechanisms of these changes were discussed.

OBJECTIVETo express and purify the protein coded by the TRAF-type zinc finger domain of myasthenia gravis (MG)-related gene P9 (P9-ZFD) and to prepare P9-ZFD antiserum for detecting expression and subcellular distribution of P9-ZFD protein in the skeletal muscles of patient with MG.

METHODSThe cDNA encoding P9-ZFD was amplified by RT-PCR. The cloned P9-ZFD cDNA was ligated into pET24a, and the P9-ZFD recombinant protein was induced via E. coli. BL21 (DE3) and purified by histidine affinity chromatography. P9-ZFD antiserum was prepared and its titer and specificity were determined by ELISA and Western blot. Expression and subcellular distribution of P9-ZFD protein in the skeletal muscles of MG and control were studied.

RESULTSThe molecular weight of purified P9-ZFD protein was about 30 kD. Its purity was more than 95%. Antiserum specific for P9-ZFD was excellent. P9-ZFD protein is fully confined to the cytoplasm membrane of skeletal muscle cell of MG, obvious immunostaining was absent in the A, I, and Z bands of cytoplasm and no immunoreactivity was observed in the skeletal muscle cell of control.

CONCLUSIONP9-ZFD protein is expressed as a cytoplasm membrane-bound protein and has obvious distribution difference in the skeletal muscle cells of patient with MG and normal control.

Adult ; Cell Membrane ; metabolism ; Escherichia coli ; metabolism ; Female ; Humans ; Muscle Proteins ; biosynthesis ; genetics ; Muscle, Skeletal ; metabolism ; pathology ; Myasthenia Gravis ; metabolism ; Peptide Fragments ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Transfection ; Zinc Fingers

Objective To investigate the role of rotational alignment reference landmarks of the proximal tibia in total knee arthroplasty.Methods Fifteen healthy adult volunteers were enrolled in this study,including 10 males and 5 females,aged from 21 to 38 years (average,28.1±6.0).CT scans of 26 knees were taken as the knees were placed in full extension.Two anteroposterior axes were drawn on the CT images:one line connected the middle of the posterior cruciate ligament insertion site and the medial edge of the patellar tendon,and another line connected the middle of the posterior cruciate ligament insertion site and the middle-medial 1/3 of the patellar tendon.The surgical epicondylar axis was also drawn on the CT images.Angles were measured between a line perpendicular to the surgical epicondylar axis and the two anteroposterior axes,and the angles were compared with the ideal tibial rotational alignment reference axis (0°).Results Angles between the line perpendicular to the surgical epicondylar axis and the line connecting the middle of the posterior cruciate ligament insertion site and the medial edge of the patellar tendon averaged 0.7°±2.8° (range,-5.1°-5.8°),there was no significant difference compared with 0°.Angles between the line perpendicular to the surgical epicondylar axis and the line connecting the middle of the posterior cruciate ligament insertion site and the middle-medial 1/3 of the patellar tendon averaged 6.9°±5.3° (range,-3.4°-14.1°),there was significant difference compared with 0°.Significant difference existed in angles between the two anteroposterior axes and the line perpendicular to the surgical epicondylar axis.Conclusion The line connecting the middle of the posterior cruciate ligament insertion site and the medial edge of the patellar tendon is a more reliable reference axis for the tibial component rotational alignment,which makes the femoral and tibial components in a more matching rotational position.

Objective:To investigate the influence of sCD80-Linker-sCD40L fusion protein on the unspecific anti- tumor immunity in vitro.Methods:Ovarian cancer SKOV3 cells were separately transfected with recombinant adenoviral vectors containing sCD80-Linker-sCD40L fusion gene,sCD80 gene,sCD40L gene or with control adenovirus.The expres- sion of the sCD80-Linker-sCD40L fusion protein,sCD80 protein and sCD40L protein in the supernatants of SKOV3 cells was determined by ELISA.Dendritic cells(DCs)were cultured with peripheral blood mononuclear cells from a patient with ovarian carcinoma.DCs and autologous T cells were co-cuhured and were exposed to different supernatants for 48 h. The allostimulatory effects of DCs on T cells were determined by mixed lymphocyte reaction(MLR).The unspecific kill- ing activities of induced T cells against SKOV3/K562 cells were measured by LDH-releasing assay.Results:ELISA assay showed that levels of the sCD80-Linker-sCD40L fusion protein,sCD80 protein and sCD40L protein in the supernatants of transfeced SKOV3 cells were 2.791 ng/ml,1.956 ng/ml and 1.407 ng/ml,respectively.The fusion protein-exposed DCs ([0.382?0.053]vs[0.167?0.028],P

Abstract: Objective　To analyze the distribution and drug resistance evolution characteristics of pathogenic bacteria of bloodstream infection in nine tertiary hospitals in Yunnan Province from 2017 to 2021, so as to provide reliable basis for rational selection of antibiotics in clinic. Methods Using the drug sensitive paper method or instrument method, the bacteria identification and drug sensitivity test were carried out in nine tertiary hospitals in different regions according to the unified technical scheme. The results were judged according to the Clinical and Laboratory Standards Institute (CLSI) breakpoint standard in 2021, and use WHONET5.6 for data statistical analysis. Results A total of 12 003 strains of pathogenic bacteria were isolated from bloodstream infection samples in the past five years, including 7 442 strains of Gram-negative bacteria (62.0%) and 4562 strains of Gram-positive bacteria (38.0%), with an increasing trend in the number of isolated strains; of these, 163 strains (1.4%) were isolated from outpatients and 11 840 strains (98.6%) were isolated from inpatients. The top three gram-negative bacteria were Escherichia coli, Klebsiella pneumoniae and Acinetobacter baumannii, of which 309 strains (4.2%) were carbapenem-resistant Klebsiella pneumoniae (CR-KPN), 29 strains (0.4%) carbapenem-resistant Escherichia coli and 19 strains (0.3%) carbapenem-resistant Enterobacter cloacae, and the number of CR-KPN was on the rise year by year. The top three Gram-positive bacteria were coagulase-negative staphylococci, Staphylococcus aureus and Enterococcus faecium, of which methicillin-resistant Staphylococcus aureus (MRSA) was detected for 213 strains, accounting for 27.7%, and decreased from 40.0% in 2017 to 23.4% in 2021, showing a downward trend year by year. No vancomycin-resistant staphylococci and enterococci were found. Conclusions The detection and composition of bloodstream infection pathogenic bacteria in multicenter have not changed much in the past five years, but each hospital has its own characteristics. The number of carbapenem resistant Enterobacteriaceae increased year by year, which should be paid more attention.

Objective To compare the diagnostic value of myocardial perfusion imaging (MPI) and 64 multi-slice spiral CT (64-MSCT) for coronary artery disease (CAD). Methods Fifty-two patients with suspected or known CAD were included in the study. Each patient underwent both stress and rest MPI,MSCT as well as conventional coronary angiography (CAG) within 1 month. The stress and rest MPI were scored by a 5-grade criteria (0 ～ 4) based on 17 coronary artery segments. The difference between summed stress and rest scores ＞ 1 was defined as myocardial ischemia. Stenosis in one main vessel or one main branch of the main vessel ≥50% was defined as myocardial ischemia by MSCT. CAG was used as the reference for comparison. Statistical analysis was performed using SPSS 13. 0 software. Kappa value was used to test the accordance of MPI and MSCT results. X2 test was used to evaluate the difference between MPI and MSCT results. Results The patient-based sensitivity, specificity, positive and negative predictive values and accuracy of MPI and MSCT for the diagnosis of CAD were 86.7% (26/30), 77.3% ( 17/22),83.9% (26/31), 81.0% ( 17/21), 82.7% (43/52) and 83.3% ( 25/30), 86.4% ( 19/22), 89.3%( 25/28), 79.2% ( 19/24), 84.6% (44/52), respectively. The vessel-based sensitivity, specificity, positive and negative predictive values and accuracy of MPI and MSCT were 74.5% (38/51), 81.0% (85/105 ), 65.5% (38/58), 86.7% ( 85/98), 78.8% ( 123/156 ) and 90.2% (46/51 ), 88.6% ( 93/105 ),79.3 % (46/58), 94.9% (93/98), 89.1% ( 139/156), respectively. There was no statistically significant difference between MPI and MSCT for either patient or lesion-based diagnosis (X2 =0.44, 0.21, both P ＞0.05 ). 96.0% (24/25) patients with both abnormal MPI and MSCT positive were valified by CAG while 83.3% (15/18) patients with both MPI and MSCT negative were excluded by CAG. Conclusions Both MPI and MSCT are reliable diagnostic modalities for CAD. They also provide complementary diagnostic value to each other.

Objective To explore the major work stresses and coping styles of psychiatric male nurses.Methods 95 psychiatric male nurses from a grade Ⅲ class A hospital in Beijing were surveyed by Nurse Job Stressors Questionnaires(NJSQ) and Simplified Coping Style Questionnaire (SCSQ).Results There was no significant difference in the total scores of JSQ among psychiatric male nurses with different position titles ( P＞0.05).However,statistical differences in the scores of work load and work time allocation between nurses-in-charge group,nurses group and nurse practitioners group[ (12.5±2.5) vs (8.7 ±3.7) vs (9.8 ±3.9) ;F=4.311,P＜0.05 ].The scores of married male nurses group in the source of work stress and work time allocation ( 10.6 ± 3.8) were higher than the score of single nurses group (7.8 ± 3.2 ) and showed significant difference (t =-3.198,P＜0.05 ).The scores of experienced male nurses group and unexperienced nurses group in the source of work stress were ( 14.9 ± 4.3 ) and ( 13.0 ± 4.3 ) respectively; the scores in work time allocation were ( 10.8 ± 3.6) and ( 8.5 ± 3.8 ) ; the scores in patients care were ( 19.3 ± 7.0) and ( 16.1± 6.8 ) ; the scores in management and relationships were ( 13.3 ± 5.5 ) and ( 10.7 ± 5.5 ).All the scores of experienced group were significantly higher than the unexperienced group ( t =- 2.230,- 3.021,- 2.229,- 2.268,respectively;P ＜ 0.05 ).There was no significant differences found between the score of coping mode and position title,years of work,marital status,education or age.Conclusions Psychiatric male nurses with different position title,work experience and marital status have more work stress.They should learn self-control,release pressure and cope with it actively.

OBJECTIVETo investigate the potential of gene therapy of rat prolactinomas mediated by adenoviral vectors with a gene encoding rat tyrosine hydroxylase.

METHODSRecombinant replication-deficient adenovirus named Ad-GFP-TH with rat TH-cDNA and control adenovirus named Ad-GFP were constructed by homologous recombination in bacterial cells. The rat pituitary prolactinoma cell line MMQ are chosen as the target cells to study the effect of gene therapy on their growth and prolactin secretion mediated by Ad-GFP-TH.

RESULTSRecombinant Ad-GFP-TH and Ad-GFP were successfully reconstructed. Transfection of MMQ cells with Ad-GFP-TH not only restrained their growth but also decreased their PRL secretion.

CONCLUSIONGene therapy may serve for a potential treatment for prolactinomas, especially invasive prolactinomas.

Adenoviridae ; genetics ; Animals ; Genetic Therapy ; Genetic Vectors ; Pituitary Neoplasms ; therapy ; Prolactinoma ; therapy ; Rats ; Recombinant Proteins ; biosynthesis ; genetics ; Transfection ; Tyrosine 3-Monooxygenase ; biosynthesis ; genetics

OBJECTIVETo investigate the influence of mutual interactions between FasL expressed by colon carcinoma cells and endogenous cytokines interleukin-18 on liver metastasis and invasion of human colon cancer cells.

METHODSUsing immunohistochemical streptavidin-biotin complex (SABC) method, the expressions of Fas receptor and Fas ligand in SW620 colon carcinoma cells and Chang liver cells were observed so as to provide morphological evidence for the functions of Fas receptor and Fas ligand. In an effort to examine the cytotoxicity of effector cells, CytoTox(96) Non-Radioactive Cytotoxicity Assay was adopted to measure the LDH releasing value after the SW620 cells were co-cultured with Chang liver cells.

RESULTSIt was shown that the Fas ligand of colon carcinoma SW620 cells was positive and the positive substances were distributed in the cell membrane and cytoplasm, and the Fas receptor of colon carcinoma SW620 cells was negative. The Fas receptor of Chang liver cells turned out to be positive and the positive substances were distributed in the cell membrane, and the Fas ligand of Chang liver cells was negative. At 6 hours after co-culture of IFN-γ-stimulated Chang liver cells with interleukin-18-stimulated (for 36 h) SW620 cells or unstimulated SW620 cells, the cytotoxicity of SW620 cells to IFN-stimulated Chang liver cells at effector-to-target ratios of 10:1, 5:1, 2.5:1 and 1.25:1 was 68.3%, 49.8%, 21.1%, 9.7% (F = 76.87, P < 0.05) and 32.7%, 21.8%, 11.1%, 6.7% (F = 7.27, P < 0.05), respectively. The non-radioactive cytotoxicity assay showed that the apoptotic rate of Chang liver cells was remarkably increased with the increase of planting concentration of SW620 after the SW620 cells were co-cultured with Chang liver cells. The cytotoxicity was significantly enhanced by interleukin-18.

CONCLUSIONThe FasL expression of human colon cancer cells may be regulated by endogenous interleukin-18 in the host microenvironment and enhance the liver colonization competence of colon cancer cells through induction of apoptosis in the Fas-expressing hepatocytes.

Apoptosis ; Cell Line, Tumor ; Cells, Cultured ; Coculture Techniques ; Colonic Neoplasms ; immunology ; metabolism ; pathology ; Cytotoxicity, Immunologic ; Fas Ligand Protein ; metabolism ; Hepatocytes ; cytology ; metabolism ; Humans ; Interferon-gamma ; pharmacology ; Interleukin-18 ; pharmacology ; L-Lactate Dehydrogenase ; metabolism ; fas Receptor ; metabolism

OBJECTIVETo establish the rat model of acute spinal cord injury, followed by aprimary study on this model with ¹H NMR based on metabonomics and to explore the metabonomics and biomarkers of spinal cord injury rat.

METHODSTwenty eight-week-old adult male SD rats of clean grade, with body weight of (200±10) g, were divided into sham operation group and model group in accordance with the law of random numbers, and every group had 10 rats. The rats of sham operation group were operated without damaging the spinal cord, and rats of model group were made an animal model of spinal cord incomplete injury according to the modified Allen's method. According to BBB score to observate the motor function of rats on the 1th, 5th, and 7th days after surgery. Postoperative spinal cord tissue was collected in order to pathologic observation at the 7th day, and the metabolic profilings of serum and spinal cord from spinal cord injury rats were studied by ¹H NMR spectroscopy.

RESULTSThe hindlimb motion of rats did not obviously change in sham operation group, there was no significant difference at each time point;and rats of model group occurred flaccid paralysis of both lower extremities, there was a significant difference at each time; there was significant differences between two groups at each time. Pathological results showed the spinal cord structure was normal with uniform innervation in shame group, while in model group, the spinal cord structure was mussy, and the neurons were decreased, with inflammatory cells and necrotic tissue. Analysis of metabonomics showed that concentration of very low density fat protein (VLDL), low density fat protein (LDL), glutamine, citric acid, dimethylglycine (DMG) in the serum and glutathione, 3-OH-butyrate, N-Acetyl-L-aspartic acid (NAA), glycerophosphocholine (GPC), glutamic acid, and ascorbate in spinal cord had significant changes(<0.05).

CONCLUSIONSThere are significant differences in metabolic profile from serum and spinal cord sample between model group and sham operation group, it conduces to explain the changes of small molecular substances in serum and spinal cord tissue after spinal cord injury, this provides the research basis for targeted research on the role of metabolic markers in patients with acute spinal cord injury.