ObjectiveTo retrospectively analyze antibiotic resistance and clinical characteristics of Klebsiella pneumoniae strains for guiding the rational use of antibiotics in the area of the Bai nationality.MethodsThe antibiotic resistance and clinical characteristics of Klebsiella pneumoniae strains were retrospective analyzed, which were isolated from specimens of inpatients in First People’s Hospital of Dali between May 2016 and May 2017.ResultsAmong the 1 342 samples of various kinds of samples, 262 strains of Klebsiella pneumoniae were isolated, with the detection rate of 19.52% (262/1342). Clinical isolated strains were mainly from the new pediatric, intensive care unit, respiratory medicine, pediatrics, and mostly from sputum specimens (78.24%, 205/262). By screening of 22 kinds of antimicrobial agents, all strains had ampicillin resistance (100.00%), while none of these strains had ertapenem resistance. Extended-spectrum β-lactamases (ESBLs) positive strains’ resistance rate was higher than ESBLs negative strains (χ2=261.992, P<0.01). There were 76 drug resistant profiles, most of which were multidrug-resistant bacteria except 116 (44.27%) strains were resistant to ampicillin antibiotics only. And the number of strains in other resistant types ranged from 1 to 16. Only one of 262 strains had amikacin resistance, two of them were resistant to imipenem and meroenan.ConclusionsThere are many multidrug-resistant bacteria in Klebsiella pneumoniae in the population of Bai nationality, and there are no extensively drug resistant bacteria and pandrug-resistant bacteria strains. The strains of carbapene-resistant antibiotics should be worthy of clinical attention.
ObjectiveTo study the distributions of virulence genes of Klebsiella pneumoniae (KP) and the distribution of hypervirulent KP (HvKP), and assess the performance of a single gene to predict HvKP.MethodsPolymerase chain reaction (PCR) method was used to analyze 12 virulence-related genes (entB, irp2, iroN, iucA, mrkD, fimH, c-rmpA, p-rmpA2, p-rmpA, wzy-K1, allS and peg-344) and drug-resistance gene blaKPC among 376 clinical KP strains collected from January 2016 to December 2018. Sequence types (ST) of KP were determined after sequencing and comparison, following the detection of 7 house-keeping genes (gapA, infB, mdh, pgi, phoE, rpoB and tonB) by PCR method. Statistical analyses were made for the distributions of virulence genes of KP and the distribution of HvKP with GraphPad Prism 8 software.ResultsAmong the 376 KP strains, the positive rates of entB, irp2, iroN, iucA, mrkD, fimH, c-rmpA, p-rmpA2, p-rmpA, wzy-K1, allS and peg-344 were 100.0%, 76.9%, 22.1%, 28.2%, 97.6%, 97.1%, 1.6%, 24.5%, 21.0%, 7.4%, 4.8% and 31.6%, respectively. The positive rates of the aforementioned virulence genes in the blaKPC-positive group (n=167) were 100.0%, 94.0%, 7.2%, 16.8%, 97.0%, 96.4%, 0.0%, 15.0%, 6.6%, 0.0%, 0.0% and 21.0%, respectively, and those in the blaKPC-negative group (n=209) were 100.0%, 63.2%, 34.0%, 37.3%, 98.1%, 97.6%, 2.9%, 32.1%, 32.5%, 13.4%, 8.6% and 40.2%, respectively; there was no statistically significant difference in entB, mrkD or fimH between the two groups (P>0.05), the positive rate of irp2 was higher in the blaKPC-positive group than that in the blaKPC-negative group (P<0.05), and the positive rates of the rest virulence-related genes were lower in the blaKPC-positive group than those in the blaKPC-negative group (P<0.05). The rate of HvKP in the blaKPC-negative group was higher than that in the blaKPC-positive group (38.3% vs. 18.0%, P<0.05). As a marker of HvKP, iucA showed high sensitivity and specificity (90.9% and 97.7%), followed by p-rmpA2 (83.6% and 100.0%) and iroN (73.6% and 99.2%). ST11 accounted for 87.4% in the blaKPC-positive group, while ST23, ST20, ST54 and ST29 were the four primary types in the blaKPC-negative group, accounting for 23.4% totally.ConclusionsDifferent virulence genes mean different distributions in KP. blaKPC-negative KP is more virulent than blaKPC-positive KP. iucA and p-rmpA2 could serve as good predicators of HvKP. Armed with extreme virulence and drug-resistance, blaKPC-positive HvKP is of great clinical concern.
ObjectiveTo investigate the effect of 24-week intradialytic progressive resistance exercise on hemoglobin and iron metabolism in maintenance hemodialysis (MHD) patients.MethodsFrom April to May 2019, 62 MHD patients were enrolled and randomly assigned into exercise group (n=31) and control group (n=31). Both groups of patients received regular routine hemodialysis, on that basis, patients in the exercise group completed intradialytic resistance exercise three times per week for 24 weeks. Each exercise included 8-10 muscle groups (grasping the grip ring with both hands, flexion and extension of the elbows and shoulders on the non-vascular side and lower limbs with sandbag), 3 sets of 15 repetitions with a rest of 1-2 min between 2 sets. Exercise began with a low load, the sandbag weight was gradually increased, and the Borg score was aimed to be 11-13 points after exercise. Hemoglobin, serum ferritin, transferrin saturation, serum creatinine, high-sensitivity C-reactive protein, urea clearance index, recombinant human erythropoietin (rHuEPO) dosage at baseline and after 24 weeks, as well as the cumulative iron supplement dose and hemoglobin variation of the two groups during the study period were evaluated.ResultsThere were 20 patients in the exercise group and 30 ones in the control group who completed the study. After 24 weeks of progressive resistance exercise, the medium (lower quartile, upper quartile) of the amount of rHuEPO in the exercise group decreased from 6 000 (6 000, 9 000) U/week to 6 000 (4 500, 7 125) U/week (Z=−2.599, P=0.009), while that in the control group had no statistically significant difference (Z=−1.340, P=0.180); there was no statistically difference in hemoglobin, hemoglobin coefficient of variation, serum ferritin, transferrin saturation, or 24-week cumulative iron supplementation between the two groups.ConclusionIntradialytic progressive resistance exercise can reduce the amount of rHuEPO in MHD patients, which is benefitial to optimizing the management of hemoglobin.
【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 review the application advancements of ATP-binding cassette (ABC) transporter in medical research.Methods Relevant literatures about the applications of ABC families in medical research were reviewed. Results ABC families mainly took roles in transporting substances across cell membrane. Some of them were useful for the prediction of drug resistance and the prognosis of malignant tumors. Others were target s for molecular researches. Their expressions or mutations might be related with the occurrence of diseases. Conclusion ABC families are very important in the diagnosis and therapy for diseases. Thus they are very promising tools for future medical research.
ObjectiveTo explore the in vitro anti-tumor effect of clove extracts (CEs) on radioresistant esophageal cancer cell KYSER and its mechanism.MethodsEthanol extracts of clove bud were prepared. Gas chromatography was used to identify the main active components of CEs. In vitro cell culture method was used to observe the effect of CEs at different concentrations on KYSER cell growth. Methyl thiazolyl tetrazolium (MTT) method was used to detect the effect of different concentration CEs on KYSER’s survival and its’ manner. Transmission electron microscope (TEM) was used to observe the changes of KYSER’s organelle microstructure after CEs treated. Transwell chamber method was used to detect the impact of CEs on KYSER’s migration ability. The apoptosis rate and cell cycle distribution of KYSER treated by CEs were quantitatively determined by flow cytometry. Clone formation experiment was used to detect the clone formation ability of KYSER treaded by CEs.ResultsThe main components of CEs were eugenol, eugenol hydrocarbon, and eugenol acetate. In vitro cell culture showed that 0.4% CEs could inhibit KYSER growth. MTT assay showed that the concentration of CEs≥0.5% could inhibit the survival of KYSER in a dose-dependent manner. TEM assay showed that after treated by 0.5% CEs, KYSER’s microvilli became shorter and wider, ribosomes in the cytoplasm decreased, mitochondria atrophied, and a large number of autophagosomes were formed. Transwell migration assay showed that relative migration rates of KYSER after treated by 0.5% CEs and 0.6% CEs were (65±4)% and (41±3)%, respectively. Compared with the control group, the differences were statistically significant (P<0.001). Flow cytometry showed that the apoptosis rates of the control group, the 0.5% CEs treated group, and the 0.6% CEs treated group were (5.63±0.50)%, (11.77±0.42)%, and (19.44±0.19)%, respectively, and the differences between the control group and the two CEs treated groups were statistically significant (P<0.001). Flow cytometry showed that the G1 phase ratios of cells in the control group, the 0.5% CEs treatment group, and the 0.6% CEs treatment group were (61.99±1.20)%, (75.38±1.50)%, and (78.81±1.00)%, respectively, and the differences between the control group and the two CEs treated groups were statistically significant (P<0.001). The clonal formation experiment showed that after 24 h of CEs treatment, the clonal formation rates of the control group, the 0.5% CEs treatment group, and the 0.6% CEs treatment group were (80.5±1.0)%, (18.1±0.8)%, and (5.0±0.5)%, respectively, and the differences between the control group and the two CEs treated groups were statistically significant (P<0.001).ConclusionCEs can exert anti-tumor effect on radioresistant esophageal cancer cells by inducing autophagy and apoptosis, promoting cell cycle arrest, inhibiting cell energy metabolism, and inhibiting migration.
摘要:目的:探讨2型糖尿病合并糖尿病足患者与胰岛素抵抗的关系。方法:205例2型糖尿病患伴糖尿病足患者作为观察组,无足部病变的糖尿病患者作为对照组,观察其体重指数、空腹血糖、胰岛素、血脂等指标,两组间进行比较并相关性分析、多元回归分析。胰岛素抵抗指数(HOMAIR)=FPG×FIns/22.5。结果:糖尿病足患者的HOMAIR显著高于无糖尿病的患者(Plt;0.05)。多元回归分析显示糖尿病病程、LDL及BMI是影响2型糖尿病足患者胰岛素抵抗的主要危险因素。结论:糖尿病足患者存在着更严重的胰岛素抵抗。Abstract: Objective: To discuss the relationship between diabetes and pedopathy of type II diabetes and insulin resistance. Methods:The diabetes type II patients were divided into group A (combined with pedopathy) and group B (without pedopathy). The blood glucose and insulin of empty stomach, BMI,Alc and lipid were detected. The insulin resistance index (HOMAIR) was calculated and compared between two groups. Results:The HOMAIR was higher in group A than that in group B (Plt;0.05).The duration of disease,LDL and BMI was positive related with diabetes pedopathy. Conclusion:The insulin resistance was more worse in pedopathy of Type II diabetes.
Hyperprolactinemia is the common clinical syndrome; the causes of hyperprolactinemia are physiological, pharmacological, and pathological, in which prolactinoma is the most common cause. In drug therapy, dopamine agonists are the first choice, but there are 10%–20% of the patients who are resistant to drug therapy. This paper mainly summarized the causes, treatments, mechanisms of drug resistance, treatment during pregnancy, and progresses in the treatment of prolactinoma, so as to provide some theoretical basis to further research of hyperprolactinemia.
Objective To dynamically study the formation of multidrug resistance(MDR) of human hepatocellular carcinoma cell SMMC-7721 induced by Adriamycin (ADM) and the role of multidrug resistance-associated protein(MRP) in its mechanisms.Methods Hepatocellular carcinoma cell SMMC-7721 was cultured in RPMI-1640 medium containing ADM with progressively increased concentration or directly cultured in medium containing different concentrations of ADM. Resistant index of drug-resistant variants of SMMC-7721 cell was determined by drawing cell dosage-reaction curves.Levels of MRP mRNA expression were detected by reverse transcription-polymerase chain reaction(RTPCR). Intracellular rubidomycin(DNR) concentration was examined by flow cytometry(FCM).Results With progressive increasing of ADM concentration in medium resistant index and levels of MRP mRNA expression were correspondingly increased but intracellular DNR concentration was markly reduced. When parental cells were directly cultured in medium containing different concentrations of ADM, the higher the ADM concentration, the higher the level of MRP mRNA expression, but intracellular DNR concentration was kept at the similar high level and most cells died. Conclusion ADM may progressively induce SMMC-7721 cell resistant to multiple chemotherapeutic drugs with reduced intracellular DNR accumulation associated with the overexpression of MRP gene.
Multidrug resistance (MDR) remains the major obstacle to the success of clinical cancer chemotherapy. P-glycoprotein (P-gp), encoded by the MDR1, is an important part with complex mechanisms associated with the MDR. In order to overcome the MDR of tumors, we in the present experimental design incorporated small interfering RNA (siRNA) targeting MDR1 gene and anticancer drug paclitaxel (PTX) into the solid lipid nanoparticles (SLNs) to achieve the combinational therapeutic effects of genetherapy and chemotherapy. In this study, siRNA-PTX-SLNs were successfully prepared. The cytotoxicity of blank SLNs and siRNA-PTX-SLNs in MCF-7 cells and MCF-7/ADR cells were detected by MTT; and the uptake efficiency of PTX in MCF-7/ADR cells were detected via HPLC method; quantitative real-time PCR and flow cytometry were performed to investigate the silencing effect of siRNA-PTX-SLNs on MDR1 gene in MCF-7/ADR cells. The results showed that PTX loaded SLNs could significantly inhibit the growth of tumor cells, and more importantly, the MDR tumor cells treated with siRNA-PTX-SLNs showed the lowest viability. HPLC study showed that SLNs could enhance the cellular uptake for PTX. Meanwhile, siRNA delivered by SLNs significantly decreased the P-gp expression in MDR tumor cells, thus increased the cellular accumulation of rhodamine123 as a P-gp substrate. In conclusion, the MDR1 gene could be silenced by siRNA-PTX-SLNs, which could promote the growth inhibition efficiency of PTX on tumor cells, leading to synergetic effect on MDR tumor therapy.