ObjectiveTo explore the prognostic risk factors of bloodstream infections caused by Acinetobacter baumannii in the hospital, to provide a basis for clinical diagnosis and treatment.MethodsA retrospective analysis was performed on the medical records of patients diagnosed with Acinetobacter baumannii bloodstream infection in Guangxi Zhuang Autonomous Region People’s Hospital between January 2013 and December 2018. The patients were divided into survival group and non-survival group according to the outcome within 30 days after blood culture was collected. Univariate and multivariate logistic analyses were used to identify the risk factors of Acinetobacter baumannii bloodstream infections.ResultsA total of 123 patients were included, including 48 in the survival group and 75 in the non-survival group. Third generation cephalosporins [odds ratio (OR)=2.492, 95% confidence interval (CI) (2.125, 2.924), P<0.001], carbapenems [OR=1.721, 95%CI (1.505, 1.969), P<0.001], multidrug resistant-Acinetobacter baumannii infection [OR=1.240, 95%CI (1.063, 1.446), P=0.006], post-operation [OR=0.515, 95%CI (0.449, 0.590), P<0.001], mechanical ventilation [OR=1.182, 95%CI (1.005, 1.388), P=0.043], indwelling central venous catheter [OR=0.116, 95%CI (0.080, 0.169), P<0.001], mixed infection or septic shock [OR=3.935, 95%CI (2.740, 5.650), P<0.001], APACHE Ⅱ score (≥15) [OR=5.939, 95%CI (5.029, 7.013), P<0.001], chronic kidney disease [OR=1.440, 95%CI (1.247, 1.662), P<0.001], immune system disease [OR=28.620, 95%CI (17.087, 47.937), P<0.001], use of corticosteroids [OR=0.520, 95%CI (0.427, 0.635), P<0.001], and combined antifungal agents [OR=0.814, 95%CI (0.668, 0.992), P=0.041] were independent factors for predicting the prognosis of patients with bloodstream infections caused by Acinetobacter baumannii.ConclusionsThe third generation cephalosporins, carbapenem, MDR-Acinetobacter baumannii infection, post-operation, mechanical ventilation, indwelling central venous catheter, mixed infection or septic shock, APACHE Ⅱ score (≥15), chronic kidney disease, immune system disease, use of corticosteroids, and combined antifungal agents were independent factors for predicting the prognosis of patients with bloodstream infections caused by Acinetobacter baumannii. In the clinical work, it is needed to carry out timely detection of microbial etiology, timely report, and reasonable treatment.
Objective To investigate the reversal effect of antisense phosphorothioate oligonucleotide （ASOND) on human hepatoma resistant cells. Methods Human hepatoma resistant cells SMMC-7721 was transfected with synthetic antisense phosphorothioate oligonucleotide complementary to the 5′ region flanking the AUG initiation codon mediated by lipofectamine. In vitro drug sensitivity was measured by MTT assay. The expression of P-170 was determined by flow cytometry and mRNA was assessed by RT-PCR. Results ASOND inhibited the expression of mRNA and p-170 in SMMC-7721, enhanced the sensitivity of SMMC-7721 to chemotherapeutic drug. The best inhibitory effect was achived by the dose of 0.5μmol/L. Conclusion ASOND enhanced the sensitivity of SMMC-7721 to chemotherapeutic drug and reversed the multidrug resistance of SMMC-7721 partially.
【Abstract】 Objective To detect the expression of lung resistance protein (LRP) and investigate its significance in pancreatic carcinoma cell lines (SW1990, PCT-2, PCT-3, PCT-4, Aspc-1, Capan-1, Mia-PaCa-2 and Panc-1). Methods Reverse transcription PCR (RT-PCR) and immunocytochemistry (ICC) were carried out to investigate the expression of LRP. Results LRP mRNA was absent in PCT-2 cell line by RT-PCR. Mild to moderate expression level was found in other pancreatic carcinoma cell lines. PCT-4, Aspc-1 and Panc-1 presented the highest LRP mRNA expression level, in contrast, SW1990, PCT-3, Capan-1 and Mia-PaCa-2 showed moderate LRP mRNA expression. The median value was 0.56±0.33. LRP was further validated by ICC. Absent to weak protein expression of LRP was found in PCT-2 and PCT-3. Overexpressed LRP was present in SW1990, Capan-1 and Aspc-1, furthermore, the highest expression of LRP was found in Panc-1, Mia-PaCa-2 and PCT-4 cell lines. Conclusion All these data showed that LRP might play an important role in multidrug resistance of pancreatic carcinoma.
Objective To review the recent studies on the multidrug resistance of breast cancer. Methods The literatures of recent years on the studies of multidrug resistance, multidrug resistance protein and breast cancer resistance protein were reviewed. Results Multidrug resistance resulted from multiple factors. How to identify the sensibility of chemotherapy drugs and select individual therapeutic regime early were important to improve the survival rate and life quality of breast cancer patients. Conclusion These studies on multidrug resistance of breast cancer are helpful to predicting the effect and outcome of chemotherapy and overcoming the barrier of drug resistance.
Objective To investigate the effect of phosphorothioate antisense oligonucleotides(AS-ODN) on suppressing multidrug resistance-associated protein gene(MRP) in human drug-resistant hepatocellular carcinoma cell line (SMMC-7721/ADM). Methods Cell line was transfected with a synthetic S-ODN complementary to the coding region of MRP mRNA, Lipofectamine acting as carrier. The drug sensitivity was measured by MTT assay. The expression of MRP mRNA was detected by RT-PCR and the expression of P190 was detected by flow cytometry. Results AS-ODN inhibited expression of MRP mRNA and P190 and promoted sensitivity to daunorubicinum and adriamycinum. Conclusion AS-ODN can reduce the expression of MRP gene. MDR caused by MRP is an important cause of multidrug resistance of SMMC-7721/ADM.
Objective To investigate the reversal of the multidrug resistant gene mdr1 in vivo by antisense oligodeoxynucleotide （ASODN） on the basis of study in vitro. Methods The cultured drug-resistant human hepatocellular carcinoma cells were injected under the skin of axilla to establish the tumor model of nude mice. mdr1 ASODN accompanied by Lipofectamine were injected locally and ADM was injected intraperitoneally. Control 1 and control 2 were locally injected by Lipofectamine and normal saline separately, and ADM was also injected intraperitoneally. Results As time went on the tumor size increased and from the 5th day on alterations were marked, tumor size in different time phase showed marked difference to the prior time phase with significant difference (P＜0.05）. Tumor size in group ASODN was marked smaller than that of other 3 groups after the 5th day (P＜0.05)，while tumor size of group control 1,2 and group SODN in different phase showed no significant difference (Pgt;0.05). The results suggested that SODN and Lipofectamine showed no marked effect on tumor growth of nude mice and ASODN had marked inhibition effect on tumor growth. Conclusion mdr1 ASODN can also reverse multidrug resistance of drug-resistant human hepatocellular carcinoma cells in vivo. After the treatment the tumor’s growth in nude mice will slow down in a range of time.
ObjectiveTo construct the recombinant adenovirus vector carrying antisense multidrug resistanceassociated protein (MRP) and transfect the human drugresistant hepatocellular carcinoma cell line(SMMC7721/ADM). MethodsThe fragment of MRP gene encoding 5′region was cloned reversely into the shuttle plasmid pAdTrackCMV, with the resultant plasmid and the backbone plasmid pAdEasy1,the homologous recombination took place in the bacteria and the recombinant adenoviral plasmid was generated. The adenoviruses were packaged and amplified in 293 cells. Then the cell line of SMMC7721/ADM was transfected with the resultant adenoviruses.ResultsThe recombinant adenovirus vector carrying antisense MRP was constructed successfully. The viral titer was 2.5×109 efu/ml, and more than 90% SMMC7721/ADM cells could be transfected when the multiplicity of infection（MOI） was 100. ConclusionThe recombinant adenovirus vector constructed by us could introduce the antisense MRP into the human drugresistant hepatocellular cell line effectively, which would provide experimental basis for the mechanisms and reversal methods of the multidrug resistance in human hepatocellular carcinoma.
Objective To study the expression and significance of multidrug resistance-associated protein (MRP) gene in hepatocellular carcinoma (HCC). Methods Reverse transcription polymerase chain reaction (RT-PCR) assay was used to detect the expression of MRP mRNA in 25 fresh specimens of the primary HCC and its surrounding liver tissues. Immunohistochemistry LSAB technique was adopted to test MRP in 60 HCC specimens. The drug sensitivity was also tested by flow cytometry.Results The positive expression rates of MRP mRNA and MRP protein in primary HCC were 44.00%(11/25) and 45.00%(27/60) respectively. All the intensity of expression was low, but significant higer than its surrouding liver tissues (P＜0.05). The intensity and expression rate of MRP protein in 5 recurrent HCC had a tendency to increase. There was a correlation between the expression of MRP mRNA and MRP protein in 25 patients using RT-PCR and immunohistochemistry assay (Plt;0.05). Detected by flow cytometry, the average sensitivity of drugs in vitro of 60 HCC sp-cimens were 5-FU （15.80±7.63）%，DDP（18.45±9.59）%，ADM（17.95±7.99）%，MMC（16.60±8.69）% and CTX（17.40±10.14）%. Only 5FU and ADM were significantly affected by the expression of MRP protein (Plt;0.05).Conclusion The expression of MRP in primary HCC may be one of the important mechanisms of the intrinsic and acquired drug resistance in HCC. To study the expression of MRP could give a predictive value in HCC chemotherapy.
【Abstract】ObjectiveTo construct a mrp1 expression vector and investigate its biological characteristics in HepG2 cells in vitro. MethodsThe 6.5 kb multidrug resistanceassociated protein （MRP) cDNA obtained from plasmid pGEM-mrp1 was cloned into the pCI-neo mammalian expression vector, which was later transferred into human hepatocarcinoma cell line HepG2 by liposome. Then the HepG2 cells resisting G418 were clustered and proliferated, and the mrp1 mRNA and MRP in these HepG2 cells were detected by means of RT-PCR and FCM respectively. ResultsThe mrp1 expression vector was established successfully, and the stable MDR hepatocarcinoma cell line (HepG2/mrp1) was developed as well. The content of the specific fragment of mrp1 mRNA was (56.8±6.37)% and MRP was 7.89 in the HepG2/mrp1 cells, the corresponding value in HepG2 cells was (9.67±3.26)% and 0.79 respectively. The difference was statistically significant (P＜0.05). ConclusionIt is practicable to establish MDR hepatocarcinoma cell line by transferring mrp1 cDNA into HepG2 cells, which is useful in the research of MDR mechanism.
Objective To detect the characteristic of multidrug resistance gene products expressions in gastric cancer tissues, including glutathione-s-transferase π (GST-π), P-glycoprotein (P-gp), topoisomerase Ⅱ (Topo-Ⅱ), thymidylate synthase (TS), and multidrug resistance related protein (MRP), and analyze their clinical significance for the therapy of gastric cancer. Methods SP immunohistochemical stain was used to detect GST-π, P-gp, Topo-Ⅱ, TS and MRP expressions in sample of 48 gastric cancer tissues and 10 normal gastric mucosa. And their corresponding clinical data were comprehensive analyzed. Results The expressions of GST-π, P-gp, Topo-Ⅱ, TS and MRP had notable differences between the gastric cancer tissues and normal gastric mucosa (GST-π： P＜0.01； P-gp： P＜0.01； Topo-Ⅱ： P＜0.01； TS： P＜0.05； MRP： P＜0.05). Positive expression rates of GST-π, P-gp, Topo-Ⅱ, TS and MRP in gastric cancer tissues were 72.9% (35/48), 56.3% (27/48), 83.3% (40/48), 41.7% (20/48) and 39.6% (19/48), and positive expression rates of them in normal gastric mucosa were 10.0% (1/10), 0 (0/10), 0 (0/10), 0 (0/10) and 0 (0/10) corresponding. Their positive expression rates were closely relevant to the degree of differentiation (P＜0.01), but not to the patients’ sex, age, tumour site, size of tumour, invasive depth and lymph node metastasis (Pgt;0.05). Conclusions The expressions of GST-π, P-gp, Topo-Ⅱ, TS and MRP in gastric cancer tissues exist obvious heterogeneity. Their overexpression underlie the multidrug resistance of gastric cancer. The joint detection of GST-π, P-gp, Topo-Ⅱ, TS and MRP can be looked as an important symbol for guiding its chemotherapy.