Objective To observe the influence of bone marrow mesenchymal stem cells (BMSCs) transplantation on the expression of alpha-amino hydroxymethyl-oxazole propionic acid (AMPA) receptors GluR1 and GluR2 in rats with spinal cord injury (SCI) so as to investigate the potential anti- chronic stress mechanism of BMSCs transplantation in treatment of rats with spinal cord injury.Methods A total of 48 adult male SD rats were equally divided into three groups:control group,treatment group and model group.The rats in the model and treatment group underwent lower thoracic SCI with the modified Allen' s method,and the rats in control group received only laminectomy.At day 7 after thoracic SCI,100 μl of Hank's buffered saline solution containing 1.0 × 106 BMSCs was injected into the subarachnoid space from L4-L5 intervertebral space in the treatment group and control group,and the same amount of Hank' s buffered saline solution was injected in the model group.The motor function of the rat posterior limbs was assessed by Basso-Beattie-Bresnahan (BBB) scale before and after operation.Half of the rats were anesthetized at days 14 and 28 postoperatively to harvest brains which were frozen and cut in a cryostat to detect the expressions of GluR1 and GluR2 proteins by immunohistochemistry.Results After BMSCs transplantation,the motor function of the posterior limbs in the treatment group was improved progressively.At day 14 after transplantation,the number of GluR1-positive cells of the model and treatment group was higher than that of the control group in the hippocampus CA1 region (P ＜0.05,P ＜0.01 respectively) ; GluR2-positive cells had the similar tendency,without significant difference(P ＞ 0.05 ).At the 28th day after transplantation,GluR1 positive cells of the model group were higher than those of the control group in CA1,CA3,DG regions and those of the treatment group in CA1,CA3 regions (P ＜0.05,P ＜0.01,respectively) ; GluR1 positive cells of the model and treatment group were higher than their counterpart at day 14 after grafting procedure,with significant difference (P ＜0.05,P ＜0.01,respectively).GluR2 positive cells of the treatment group were higher than those of the control group in the basolateral amygdale (BLA) (P ＜0.05 ) and had similar tendency with GluR1 expression in other regions ( P ＞ 0.05 ).Conclusion BMSCs transplantation implies a potential antichronic stress mechanism of SCI rats,since it can improve the motor function of posterior limbs in rats with lower thoracic SCI and regulate the expressions of AMPA receptor GluR1 and GluR2.

Objective To investigate the clinical efficacy of fluoroscopy-guided intestinal adhesion lysis, as a new non-surgical method, in treating incomplete adhesive small intestinal obstruction in order to improve the therapeutic results of adhesive intestinal obstruction. Methods A total of 93 patients with incomplete adhesive small intestinal obstruction were enrolled in this study. The patients were divided into study group (n=49) and control group (n=44). Fluoroscopy-guided intestinal adhesion lysis together with restoration of inter-intestinal loop enterocele was carried out for the patients of the study group , while traditional conservative surgical therapy was employed for the patients of the control group. The study group was comparable with the control group in patients’ age, gender, medical history, disease course, X-ray findings, etc. Results Of the 49 cases in the study group, complete cure was obtained in 40 with a cure rate of 81.6%. The mean hospitalization day was 0.3 day, and the average operation time was 3.25 hours. Among the 44 patients in the control group, complete cure was obtained in 37 with a cure rate of 84.1%. The mean hospitalization day was 7.6 days, and the average therapeutic time was 183.26 hours. Conclusion For the treatment of incomplete adhesive small intestinal obstruction , the therapeutic efficacy of fluoroscopy-guided intestinal adhesion lysis together with restoration of inter-intestinal loop enterocele is better than that of traditional conservative surgical therapy.

Abstract: Objective To investigate the clinical distribution characteristics, drug resistance trends and the carrying of antiseptic resistance gene of Pseudomonas aeruginosa infection in children in Suzhou, in order to provide theoretical basis for the prevention and treatment of Pseudomonas aeruginosa infection in children. Methods The clinical distribution characteristics and drug resistance trends of Pseudomonas aeruginosa isolated from Children's Hospital of Soochow University from 2016 to 2021 were retrospectively analyzed. Forthermore, 101 strains of Pseudomonas aeruginosa were randomly selected to detect the expression of 9 antiseptic resistance genes (qacEΔ1-sul1, qacE, qacEΔ1, qacG, sugE(p), sugE©, emrE, ydgE, ydgF) by polymerase chain reaction. Results Pseudomonas aeruginosa in Soochow University Children's Hospital was mainly isolated from respiratory specimen (47.83%), pus (28.60%) and urine (11.72%); the main departments were intensive care unit(21.45%), general surgery department (15.71%) and respiratory department (12.31%). Patients were mainly aged from 1 month to 1 year old and older than 6 years old (34.31% and 25.38%). The top three drug resistance rates of Pseudomonas aeruginosa were imipenem (11.25%), aztreonam (9.26%) and meropenem (8.02%). Among the 853 strains of Pseudomonas aeruginosa, the drug-resistant strains were mainly from the intensive care unit (58/183), hematology department (33/91), neonatology department (31/96), and there were 57 strains of multi-drug-resistant strains with the detection rate of 6.68%. There were 98 strains (11.49%) of Carbapenem resistant Pseudomonas aeruginosa, and the annual detection rates were 22.06%, 8.40%, 3.60%, 5.67%, 9.85% and 17.20%, respectively. Among the 9 antiseptic resistance genes, the carrying rate of ydgF, sugE© and qacE was 98.02%, 94.06% and 0 respectively. Conclusion Pseudomonas aeruginosa has high resistance to some drugs, so attention should be paid to rational drug use. The carriage rates of of two antiseptic resistance genes exceeded 90%, indicating the need to strengthen research on the mechanism of antiseptic resistance research and rational use of disinfectants

OBJECTIVE: LncRNA ITGB2-AS1 has been found to play important roles in the occurrence and development of human solid tumors. However, its role in hematological diseases, especially acute myeloid leukemia (AML), remains unclear. The aim of this study was to identify the expression pattern of ITGB2-AS1 in AML patients and to further explore its clinical significance. METHODS: ITGB2-AS1 expression was analyzed in public datasets (including TCGA and GSE63270) and further validated in a cohort of 109 AML patients by real-time quantitative PCR (RT-qPCR). RESULTS: The level of ITGB2-AS1 was up-regulated among two independent cohorts (TCGA, P<0.05; GSE63270, P<0.05), which was confirmed by the data from 109 AML patients enrolled in this study (P<0.05). Clinically, high ITGB2-AS1 expression was associated with older age (P=0.023) and lower complete remission (CR) rate (P=0.005). Multivariate analysis identified that high ITGB2-AS1 expression was an independent prognostic factor not only for CR rate (P=0.027) but also for overall survival (OS) time (P=0.011), and ITGB2-AS1 was positively correlated with ITGB2 expression in both TCGA (r=0.74, P<0.001) and clinical data detected in this study (r=0.881, P<0.001). High ITGB2 expression was also associated with older age (P=0.02) and lower CR rate (P=0.020). Moreover, high ITGB2 expression predicted worse OS (P=0.028). CONCLUSION ITGB2-AS1 is overexpressed in AML and predicts poor prognosis in AML patients.
Aged ; Humans ; Leukemia, Myeloid, Acute/genetics* ; Prognosis ; RNA, Long Noncoding/genetics*

OBJECTIVETo investigate the reversal effect of targeted modulation of bcl-2 expression by miR-15a and miR-16 on drug resistance of human colon cancer cells.

METHODSMimics or inhibitors of miR-15a and miR-16 were transfected into HCT8 or HCT8/VCR cells with the help of Lipofectamine 2000. The expressions of miR-15a and miR-16 mRNA were detected by RT-qPCR. The levels of bcl-2 and P-gp proteins were measured by Western blot. The inhibitory effects of VCR on growth of HCT8 and HCT8/VCR cells were detected by CCK8.

RESULTSAfter transfection of the mimics, the expression of miR-15a in the blank control group, negative control group and miR-15a mimic group was 1.00, 0.87 ± 0.24, and 223.44 ± 59.07, respectively, and miR-15a was increased significantly (P < 0.001). The expression of miR-16 in the blank control group, negative control group and miR-16 mimic group was 1.00, 0.66 ± 0.19, and 107.32 ± 22.58, respectively, and miR-16 expression was increased significantly (P < 0.001). The Western blot assay showed that the relative expressions of bcl-2 protein in the blank control group, negative control group, miR-15a mimic group and miR-16 mimic group were 1.00, 0.97 ± 0.02, 0.51 ± 0.06, and 0.65 ± 0.03, respectively, and the expression of bcl-2 protein was decreased significantly (P < 0.05), however, the expressions of P-gp protein showed no significant difference. The CCK8 test showed that at 1, 5, 25 and 125 µg/ml concentration of VCR, the survival rates of HCT8/VCR cells were basically the same in the blank control group, negative control group, miR-15a mimic group and miR-16 mimic group, but the survival rate of HCT8/VCR cells was significantly decreased after transfection of mimics (P < 0.05). After transfection of the inhibitors, the expressions of both miR-15a and miR-16 were decreased significantly (P < 0.001). The Western blot showed that the expression of bcl-2 protein was increased (P < 0.05), while the expression of P-gp protein showed no significant difference. The CCK8 test showed that the survival rate of HCT8 cells which were transfected with inhibitors was significantly higher than that of the blank control group (P < 0.05).

CONCLUSIONSmiR-15a and miR-16 may reverse the drug resistance in human colon cancer cells. A possible mechanism is regulating the expression of bcl-2.

ATP-Binding Cassette, Sub-Family B, Member 1 ; Apoptosis ; Apoptosis Regulatory Proteins ; metabolism ; Cell Line, Tumor ; Colonic Neoplasms ; Drug Resistance ; Humans ; MicroRNAs ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; RNA, Messenger ; Transfection

OBJECTIVEA subcutaneous transplantation tumor model of human HT-29 cells in nude mice was established to evaluate anticarcinogenic activities, and the apoptosis-regulated mechanism effect of aqueous extract of fermented wheat germ with Lactobacillus plantarum dy-1 (LFWGE).

METHODSThe HT-29 cells were transplanted via subcutaneous injection of 1×107 cells into the right flank of each nude mouse. Then, nude mice were treated for 30 d with LFWGE (high-dose 2 g/kg/d; low-dose 1 g/kg/d) and for 7 d with 5-fluorouracil (5-FU, 25 mg/kg/d) by gavage and intraperitoneal injection, respectively. An inhibition of tumor growth was observed.

RESULTSTumor volume and weights decreased significantly in both groups of nude mice treated with LFWGE. In addition, the cell apoptosis rate of the LFWGE group (2 g/kg/d, 60.1%±4.4%; 1 g/kg/d, 58.6%±6.9%) was significantly higher than that of the control group (11.5%±1.6%) and 5-FU group (32.1%±3.5%) as measured by the TUNEL assay. Moreover, the real-time fluorescent quantitative PCR and Western blot method further confirmed these enhancing apoptosis and growth inhibition effects. The involvement of LFWGE in inducing apoptosis was confirmed by the expression of Bax, Bcl-2, Caspase-3, and CyclinD1.

CONCLUSIONThe results showed that LFWGE could induce subcutaneous transplantation tumor apoptosis in nude mice and could be as a natural nutrient supplements or chemopreventive agent in the treatment of human colon cancer.

Animals ; Antineoplastic Agents ; chemistry ; pharmacology ; Apoptosis ; drug effects ; Caspase 3 ; genetics ; metabolism ; Cyclin D1 ; genetics ; metabolism ; Gene Expression Regulation, Neoplastic ; drug effects ; HT29 Cells ; Humans ; Mice ; Mice, Nude ; Neoplasms, Experimental ; drug therapy ; Plant Extracts ; chemistry ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; bcl-2-Associated X Protein ; genetics ; metabolism

This study is to explore the interventional effects of fluvastatin on anti-beta2GPI/beta2GPI-induced activation in THP-1 mononuclear cells. In vitro, human mononuclear cells THP-1 were treated with fluvastatin, LPS and anti-beta2GPI/beta2GPI, then the TF expression on THP-1 cells was detected by real-time quantitative PCR (RT-qPCR) or TF activity was detected by kit. TNF-alpha mRNA and its protein expression were investigated by RT-PCR and ELISA kit. The expression of phospho-NF-kappaB p65 and inhibitory protein of NF-kappaB (IkappaB-alpha) were measured by Western blotting. The results suggested that the expression of TF and TNF-alpha on THP-1 cells was significantly up-regulated with treatment of anti-beta2GPI/beta2GPI complex (100 mg x L(-1)), compared with that of untreated cells (P < 0.05). Fluvastatin (50 mg x L(-1)) could decrease TF (mRNA and activity) expression and the level of TNF-alpha (mRNA and protein) in THP-1 cells with anti-beta2GPI/beta2GPI complex. The expression of TF and TNF-alpha was shown in a concentration-dependent manner. Moreover, anti-beta2GPI/beta2GPI complex could downregulate IkappaB-alpha levels and increase the levels of phospho-NF-kappaB p65. And these effects of anti-beta2GPI/beta2GPI complex could be blocked by fluvastatin. In conclusion, fluvastatin may interfere the expression and regulation of NF-kappaB signal transduction pathway, thereby inhibit the effects of anti-beta2GPI/beta2GPI on activation of THP-1 cells, by decreasing the expression of TF and TNF-alpha.
Antigen-Antibody Complex ; pharmacology ; Cell Line ; Dose-Response Relationship, Drug ; Fatty Acids, Monounsaturated ; administration & dosage ; pharmacology ; Humans ; Hydroxymethylglutaryl-CoA Reductase Inhibitors ; administration & dosage ; pharmacology ; I-kappa B Proteins ; metabolism ; Indoles ; administration & dosage ; pharmacology ; Monocytes ; cytology ; metabolism ; NF-KappaB Inhibitor alpha ; Phosphorylation ; RNA, Messenger ; metabolism ; Signal Transduction ; drug effects ; Thromboplastin ; genetics ; metabolism ; Transcription Factor RelA ; metabolism ; Tumor Necrosis Factor-alpha ; genetics ; metabolism ; beta 2-Glycoprotein I ; antagonists & inhibitors ; immunology

Objective To investigate the quinolone resistance determinants in ciprofloxacin-resistant Acinetobacter bau-mannii (ABA)clinical isolates.Methods One hundred and fourteen ciprofloxacin-resistant ABA strains were collected from six Chinese hospitals .The quinolone resistance determining region ( QRDR) of 4 target genes ( gyrA, gyrB, parC and parE) was amplified , sequenced and compared with the reference genome of ATCC 17978 to identify possible resistance-related mutations.Nine plasmid-mediated quinolone resistance (PMQR) genes (qnrA, qnrB, qnrC, qnrD, qnrS, qepA, aac(6′)-Ⅰb-cr, oqxA and oqxB) were also amplified, and the amplicons were then sequenced to determine their character-istics.Results Almost all isolates (113/114, 99.1%) harbored a substitution in codon 83 of gyrA gene, leading to a Ser83Leu mutation.Meanwhile,58.8%(67/114) of the isolates possessed dual mutations of GyrA-Ser83Leu and GyrA-Ser80Leu, which were known determinants for ciprofloxacin resistance .There were also multiple non-synonymous substitu-tions in gyrB, leading to Arg393Ser, Arg393Cys, Thr401Ala, Pro406Ser, Val430Phe, Cys440Ser and Gly480Arg muta-tions with prevalence rates of 95.6%, 0.9%, 96.5%, 96.5%, 100%, 96.5%and 96.5%,respectively.For parE, all the seven mutations were synonymous and found in more than 96%of the tested isolates.For PMQR genes, although 83.3%(95/114) of the isolates were positive for aac(6′)-Ⅰb, nocrmutations were identified.None of the other eight PMDR genes were found in our strain collection .Conclusion Although multiple mutations are identified in gyrB and parE, these mutations might be the characteristic SNP markers for specific clones , unlikely linked to quinolone resistance .No PMQR is found in the tested isolates.Mutations in chromosomal QRDR (GyrA-Ser83Leu and ParC-Ser80Leu) are the main determi-nants of ciprofloxacin resistance in our ABA collection .

This study was aimed to investigate the effect of PTD-mFoxp3 fusion protein on graft-versus-host disease (GVHD) after allogeneic bone marrow transplantation. The 10-weeks-old C57BL/6 mice as recipients were randomly divided into three groups (A,B and C), 10 mice were in each group. The mice on day of transplantation as on day 0 received total body irradiation (TBI) 6.0 Gy, then the bone marrow cells (BMC) from BALB/c mice were injected through tail vein within 4-6 hours. At 2 days before transplantation and 0, 1, 3, 5, 7, 9 and 13 days after transplantation, mice in group A were injected with saline, mice in group B were injected with mFoxp3 protein and mice in group C were injected with PTD-mFoxp3 fusion protein. Symptoms of GVHD, survival time and histopathological changes were observed. The establishment of mixed chimerism was determined by flow cytometry in day 60, and IL-2 and IFN-γ expression profiles in the recipient peripheral blood were assessed by ELISA. The results showed that the mean survival time of recipients in group A,B and C was (32.95 ± 5.48) , (38.00 ± 5.45) and (55.30 ± 3.15) respectively. Graft rejection was observed in the liver and small intestine specimens of group A and group B. The serum levels of IL-2 and IFN-γ significantly decreased in the recipients of group C, as compared with the other groups. The flow cytometry analysis revealed that the survival recipient mice developed high chimerism levels, the percentages of donor cells in group A,B and C were (79.46 ± 1.80) %, (79.13 ± 2.23) % and (85.92 ± 2.82) % respectively. It is concluded that PTD-mFoxp3 fusion protein can reduce the incidence and mortality of GVHD after allogeneic bone marrow transplantation.
Animals ; Bone Marrow Transplantation ; adverse effects ; Female ; Forkhead Transcription Factors ; therapeutic use ; Graft vs Host Disease ; metabolism ; therapy ; Interferon-gamma ; blood ; Interleukin-2 ; blood ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Recombinant Fusion Proteins ; therapeutic use ; Transplantation, Homologous