[Risk factors for delirium after sedation in children with convulsion and establishment of a nomogram model for predicting the risk of delirium]. / æƒŠåŽ¥æ‚£å„¿é•‡é™åŽè°µå¦„å‘ç”Ÿçš„å±é™©å› ç´ åŠé£Žé™©åˆ—çº¿å›¾é¢„æµ‹æ¨¡åž‹çš„å»ºç«‹.

OBJECTIVES: To investigate the risk factors for delirium after sedation in children with convulsion, and to establish a nomogram model for predicting the risk of delirium. METHODS: A total of 373 children with convulsion who were hospitalized in the pediatric ward of the Second Affiliated Hospital of Air Force Medical University from August 2020 to January 2022 were prospectively enrolled. There were 245 children in the modeling group and 128 children in the validation group. A multivariate logistic regression analysis was used to identify independent predictive factors for delirium after sedation and establish a nomogram model for predicting the risk of this disorder based on these factors. The calibration curve, the receiver operating characteristic curve, and the decision curve analysis were used to evaluate the accuracy, discriminatory ability, and clinical application value of this model, respectively. RESULTS: The incidence of delirium after sedation was 22.3% (83/373) in the children with convulsion. The multivariate logistic regression analysis showed that age>5 years (OR=0.401, P<0.05) was a protective factor against delirium after sedation in these children, while presence of infection (OR=3.020, P<0.05), admission to the pediatric intensive care unit (OR=3.126, P<0.05), use of benzodiazepines (OR=5.219, P<0.05), history of status convulsion (OR=2.623, P<0.05), and history of delirium episodes (OR=3.119, P<0.05) were risk factors for delirium. The H-L deviation test of the nomogram prediction model showed a good degree of fit (χ2=9.494, P=0.302). Internal and external validation showed that the mean absolute errors between the actual and predicted values of the calibration curve were 0.030 and 0.018, respectively, and the areas under the receiver operating characteristic curve were 0.777 and 0.775, respectively. The decision curve analysis showed that the model provided significant net clinical benefit when the predicted risk threshold was >0.01. CONCLUSIONS: Age, presence of infection, admission to the pediatric intensive care unit, use of benzodiazepines, history of status convulsion, and history of delirium episodes are closely associated with the development of delirium after sedation in children with convulsion. The nomogram model for predicting this disorder that is established based on these factors has relatively high accuracy, discriminatory ability, and clinical application value.

[Apoptosis of Burkitt's lymphoma Raji cell line induced by bortezomib].

The aim of this study was to clarify whether bortezomib might induce apoptosis in Burkitt's lymphoma Raji cell line and its mechanism. Different concentrations of bortezomib were used to treat Raji cells and its effects of time and dose were observed. Cell morphology was observed under light microscope; flow cytometry was used to analyze cell apoptosis; RT-PCR was used to detect the expressions of NF-kappaB and p53 gene mRNAs. The results showed that the bortezomib could inhibit Raji cell growth within a certain range of treating time and dose. Apoptosis were induced in relation to time and dose. The expression of NF-kappaB mRNA and p53 mRNA decreased after treatment with bortezomib. It is concluded that the bortezomib can induce Raji cell apoptosis, which provides a theoretical basis for clinical treatment. NF-kappaB and p53 gene are supposed to participate in the bortezomib induced apoptosis of Raji cells.

[Effect of ribosomal protein L6 on drug resistance and apoptosis in K562/A02 cells].

The objective of study was to investigate the effect of ribosomal protein L6 (RPL6) gene expression on the drug resistance of leukemia cells and its possible mechanism. RPL6 cDNA was obtained by RT-PCR, both sense and antisense cDNA recombinants of RPL6-encoding gene were constructed with pcDNA3. 1 (+) expression vector. Subsequently, the sense RPL6 cDNA recombinant was transfected into K562 cells while the antisense one into K562/A02 cells by liposomal reagent. The chemosensitivity, apoptosis and caspase-3 activity of K562 and K562/A02 cells were evaluated by MTT assay, flow cytometer and fluorometer respectively. The results indicated that expression of RPL6 in K562/A02 was higher than that in K562; resistance of sense-transfected K562 cells to doxorubicin was 325% of control cells, apoptosis and caspase-3 activity decreased (P<0.005); whereas resistance of antisense-transfected K562/A02 cells to adriamycin was 38% of control cells, apoptosis and caspase-3 activity significantly increased (P<0.005). It is concluded that RPL6 gene plays an important role in the development of drug resistance in K562/A02 cells by changing drug-induced apoptosis.

[Reversal of multidrug resistance by MDR1 shRNA expression vector in human leukemia K562/A02 cells].

OBJECTIVE: To construct a short hairpin RNA (shRNA) eukaryotic expression vector specific to MDR1 gene in multidrug resistance (MDR) human leukemia cell line K562/A02 to observe its silencing effect on MDR1 and P-glycprotein (P-gp) expression. METHODS: The shRNA expression vector was constructed by gene recombination, then transfected into the cultured K562/A02 cells. The transcription of MDR1 gene was detected by semi-quantitative RT-PCR and the expression level of P-gp was determined by Western blot. 50% inhibition concentration (IC50) of ADM in K562/A02 cells was determined by MTT method. The intracellular doxorubicin (ADM) concentration was determined by HPLC. RESULTS: The introduction of pEGFP-C1/U6/MDR1-A or pEGFP-C1/U6/MDR1-B expression vector was shown to efficiently and specifically inhibit the expression of P-gp according to results of Western blot, with an inhibitory rate of 50.67%. Semi-quantitative RT-PCR showed that mRNA transcription of MDR1 gene was reduced by (48.2 +/- 2.5)%. On the contrast, the control plasmid did not exhibit inhibitory effect on the protein expression and mRNA transcription of MDR1. The relative efficiency of K562/A02 to ADM was 40.8% or 62.4%, respectively, and the intracellular accumulation of ADM increased after shRNA treatment. CONCLUSION: The shRNA expression vector targeting MDR1 gene showed dramatic inhibition on RNA transcription and protein expression. It could effectively restore the sensitivity of K562/A02 cells to conventional chemotherapeutic agents.

[Construction of pEGFP-C1/U6-mediated plasmid expressing MDR1 shRNA].

To construct a plasmid expressing MDR1 short hairpin RNA (shRNA) mediated by pEGFP-C1/U6 vector, two coding sequences of 19 nucleotides were selected from MDR. Two pairs of oligonucleotides were designed for these two fragments. After annealing the formed double-stranded DNAs were ligated with plasmid pEGFP-C1/U6 (pEGFP-C1 vector with U6 promoter). The plasmids producing MDR1 shRNA were constructed from the inverted motif containing 9 spacers and four Ts. The results showed that the constructed plasmids were named pEGFP-C1/U6/A and pEGFP-C1/U6/B, and the constructs were identified by restriction and sequence analysis, no any base mutation was observed. It is concluded that plasmids of pEGFP-C1/U6/A and pEGFP-C1/U6/B expressing MDR1 shRNA were successfully constructed, providing a highly effective method for reversing the multidrug resistance in clinic.

[Effect of neferine on the chemotherapic sensitivity of STI 571 to K562/A02 cells].

OBJECTIVE: To explore the effect of neferine on the chemotherapic sensitivity of STI 571 to K562/A02 cells and to reveal its mechanism. METHODS: MTT method was used to observe the alteration of the proliferation of K562/A02 cell line treated with STI571 alone or combined with neferine. The transcription of mdrl gene was detected by semi-quantitative RT-PCR and the P-gp expression was determined by Western blot after STI 571 alone or combined with neferine treatment. RESULTS: The cytotoxic effect of STI 571 (1 micromol/L) combined with neferine (IC50 = 3.02 micromol/L) on K562/A02 cell line was significantly higher than that of STI 571 alone (IC50 = 0.689 micromol/L). After treating with STI571 combined with neferine, the synergistic interaction on K562/A02 cells increased 4.38 folds (P < 0.05); the mdrl mRNA expression by semi-quantitative RT-PCR was significantly reduced by (45.4 +/- 2.5) % (P < 0.01); and the P-gp expression by Western blot was deregulated by 40.58% (P < 0.05). CONCLUSION: Neferine significantly enhances the antineoplastic effect of STI 571 on K562/A02 cells by reducing mdrl mRNA transcription and blocking P-gp expression.

[Detection of glutathione S-transferase and lung resistance-related proteins in acute leukemia and its clinical significance].

OBJECTIVE: To explore the relationship among intracellular glutathione S-transferase activity (GST), the expression of lung resistance-related proteins (LRP) in acute leukemia, and its clinical effects. METHODS: The GST activity of bone marrow mononuclear cells and LRP expression in 57 acute leukemia patients were detected by the spectrophotometry assay and immuno-cytochemistry (SABC), respectively. RESULTS: The GST activity of bone marrow mononuclear cells in the acute leukemia group was significantly higher than that of the control group (P < 0.01). The GST activity of mononuclear cells in acute leukemia was positively correlated with the percentage of blast in the bone marrow (r = 0.30, P < 0.05). The GST activity of mononuclear cells in the untreated acute leukemia group was obviously higher than that of the complete remission group (P <0.01). The GST activity in the refractory or relapsed acute leukemia group was significantly higher than that of the complete remission group and untreated leukemia group (P <0.05). In post-chemotherapy 13 of 17 the LRP-positive patients were the non-remission, 12 of the 20 LRP-negative patients were the complete remission. The curative rate of the LRP-positive group was the significantly lower than the LRP-negative group (P < 0.05). The GST activities of non-remission patients in the LRP-positive and LRP-negative group obviously increased. CONCLUSION: The increase of GST activity in the bone marrow mononuclear cells is related to the clinical curative effects and the proliferation of blast in acute leukemia. Detection of LRP and GST activities in acute leukemia may have a reference value in judging the leukemia with drug resistance and estimating the prognosis.

[Nitric oxide synthase expression and multiple drug resistance in patients with acute leukemia].

OBJECTIVE: To determine the relationship between nitric oxide synthase (NOS) expression and Pgp/mdr-1. METHODS: The expressions of iNOS, eNOS, and Pgp were detected by immunohistochemistry, and the expressions of iNOS gene and mdr-1 gene were detected by RT-PCR. RESULTS: NOS expression increased in patients with acute leukemia compared with that of the controls. There was some correlation between iNOS and Pgp/mdr-1 gene expression. CONCLUSION: NO may influcence multiple drug resistance in patients with acute leukemia.

[Antibacterial activity of Chinese medicine kou-kang mouthwash and its acute toxicity test].

OBJECTIVE: To study the antibacterial activity of Chinese medicine kou-Kang mouthwash and its acute toxicity, and provide the basis of experiment and therapy for infectious diseases of the oral cavity. METHODS: Minimal inhibitory concentrations (MIC) of kou-Kang mouthwash were determined by the agar dilution method, and the acute toxicity was done. RESULTS: The range of MIC kou-Kang mouthwash against 27 Gram negative bacterial were 0.03-1.0 g.ml-1. MIC50 was 0.125 g.ml-1 and MIC90 was 0.125-0.5 g.ml-1. The range of MIC kou-Kang mouthwash against 21 Gram positive bacterial was 0.008-0.5 g.ml-1. The range of MIC50 and MIC90 were 0.015-0.125 g.ml-1 and 0.015-0.5 g.ml-1, respectively. The value of MIC was 0.015 g.ml-1 against candidia albicans. The acute toxic test indicated that the maximum tolerance dose was 417.6 g.kg-1 and 210 times as much as the clinical dose a day. No death of animals or toxicity and side-effect occurred in the test. CONCLUSION: Kou-Kang mouthwash shows a potent antibacterial activity against various pathogens of the oral cavity. Its antibiotic activity against Gram positive bacteria was 2-8 times stronger than that against Gram negative bacteria. It has low toxicity and can be used in clinical application.

[Expression of multidrug resistance P-170 and MRP and their correlation with clinical drug resistance in acute leukemia].

OBJECTIVE: To investigate the relevance between the expression of P-170 and MRP and clinical drug resistance in acute leukemia. METHODS: The expression of P-170 and MRP in mononuclear cells of bone marrows was analyzed by the immunohistochemical technique in 72 acute leukemia patients. RESULTS: The expression of P-170 was positive in 46 and negative in 26 of the 72 post-chemotherapy acute leukemia patients. The therapeutic effect of the P-170 positive expression patients was significantly poorer than that of the negative expression patients (P < 0.01). The expression of MRP was positive in 39 and negative in 33 of the 72 post-chemotherapy acute leukemia patients. The therapeutic effect of the MRP positive expression patients was significantly poorer than that in the negative expression patients (P < 0.01). The expression of P-170 and MRP had a significant concordance (Kappa = 0.427, P < 0.01). The sensitivity of P-170 and MRP which were analyzed simultaneously was 97.5%, which was higher than that of P-170 (90%) or MRP (77.5%) analyzed respectively in drug resistance patients. CONCLUSION: The expression of P-170 and/or MRP was significantly related with drug resistance in clinical chemotherapy. The therapeutic effect was significantly poorer in P-170 and/or MRP positive expression patients than that in negative expression patients. These data suggest that P-170 and MRP analyzed simultaneously can improve the value of diagnosis and prognosis in patients with drug resistance leukemia.