[A multicentric study on clinical characteristics and antibiotic sensitivity in children with methicillin-resistant Staphylococcus aureus infection].

Objective: To investigate the clinical characteristics of pediatric methicillin-resistant Staphylococcus aureus (MRSA) infection and the antibiotic sensitivity of the isolates. Methods: The clinical data of children with MRSA infection and antibiotic sensitivity of the isolates from 11 children's hospitals in Infectious Diseases Surveillance of Paediatrics (ISPED) group of China between January 1, 2018 and December 31, 2018 were collected retrospectively. The children's general condition, high-risk factors, antimicrobial therapy and prognosis, differences in clinical disease and laboratory test results between different age groups, and differences of antibiotic sensitivity between community-acquired (CA)-MRSA and hospital-acquired (HA)-MRSA were analyzed. The t test and Wilcoxon rank sum test were used for statistical analysis of the quantitative data and Chi-square test were used for comparison of rates. Results: Among the 452 patients, 264 were males and 188 were females, aged from 2 days to 17 years. There were 233 cases (51.5%) in the ≤1 year old group, 79 cases (17.5%) in the>1-3 years old group, 29 cases (6.4%) in the >3-5 years old group, 65 cases (14.4%) in the >5-10 years old group, and 46 cases (10.2%) in the>10 years old group. The main distributions of onset seasons were 55 cases (12.2%) in December, 47 cases (10.4%) in February, 46 cases (10.2%) in November, 45 cases (10.0%) in January, 40 cases (8.8%) in March. There were 335 cases (74.1%) CA-MRSA and 117 (25.9%) cases HA-MRSA. Among all cases, 174 cases (38.5%) had basic diseases or long-term use of hormone and immunosuppressive drugs. During the period of hospitalization, 209 cases (46.2%) received medical interventions. There were 182 patients (40.3%) had used antibiotics (ß-lactams, glycopeptides, macrolides, carbapenems, oxazolones, sulfonamides etc) 3 months before admission. The most common clinical disease was pneumonia (203 cases), followed by skin soft-tissue infection (133 cases), sepsis (92 cases), deep tissue abscess (42 cases), osteomyelitis (40 cases), and septic arthritis (26 cases), suppurative meningitis (10 cases). The proportion of pneumonia in the ≤1 year old group was higher than the >1-3 years old group,>3-5 years old group,>5-10 years old group,>10 years old group (57.5% (134/233) vs. 30.4% (24/79), 31.0% (9/29), 38.5% (25/65), 23.9% (11/46), χ(2)=17.374, 7.293, 7.410, 17.373, all P<0.01) The proportion of skin and soft tissue infections caused by CA-MRSA infection was higher than HA-MRSA (33.4% (112/335) vs. 17.9% (21/117), χ(2)=10.010, P=0.002), and the proportion of pneumonia caused by HA-MRSA infection was higher than CA-MRSA (53.0% (62/117) vs. 42.1% (141/335), χ(2)=4.166, P=0.041). The first white blood cell count of the ≤1 year old group was higher than that children > 1 year old ((15±8)×10(9)/L vs. (13±7)×10(9)/L, t=2.697, P=0.007), while the C-reactive protein of the ≤1 year old group was lower than the 1-3 years old group,>5-10 years old group,>10 years old group (8.00 (0.04-194.00) vs.17.00 (0.50-316.00), 15.20 (0.23-312.00), 21.79(0.13-219.00) mg/L, Z=3.207, 2.044, 2.513, all P<0.05), there were no significant differences in procalcitonin (PCT) between different age groups (all P>0.05). After the treatment, 131 cases were cured, 278 cases were improved, 21 cases were not cured, 12 cases died, and 10 cases were abandoned. The 452 MRSA isolates were all sensitive to vancomycin (100.0%), linezolid (100.0%), 100.0% resistant to penicillin, highly resistant to erythromycin (85.0%, 375/441), clindamycin (67.7%, 294/434), less resistant to sulfonamides (5.9%, 23/391), levofloxacin (4.5%, 19/423), gentamicin (3.2%, 14/438), rifampicin (1.8%, 8/440), minocycline (1.1%, 1/91). The antimicrobial resistance rates were not significantly different between the CA-MRSA and HA-MRSA groups (all P>0.05). Conclusions: The infection of MRSA is mainly found in infants under 3 years old. The prevalent seasons are winter and spring, and MRSA is mainly acquired in the community. The main clinical diseases are pneumonia, skin soft-tissue infection and sepsis. No MRSA isolate is resistant to vancomycin, linezolid. MRSA isolates are generally sensitive to sulfonamides, levofloxacin, gentamicin, rifampicin, minocycline, and were highly resistant to erythromycin and clindamycin. To achieve better prognosis. clinicians should initiate anti-infective treatment for children with MRSA infection according to the clinical characteristics of patients and drug sensitivity of the isolates timely and effectively.

Aclidinium bromide inhibits proliferation of osteosarcoma cells through regulation of PI3K/Akt pathway.

OBJECTIVE: Osteosarcoma is recognized as the most common primary malignant bone tumor, the 5-year disease-free survival rate in patients with metastatic or recurrent disease is below than 30%. Drug resistance and toxic side effects limit the therapeutic efficacy of osteosarcoma. Therefore, it is urgent to develop new drugs for osteosarcoma treatment. Muscarinic 3 (M3) acetylcholine receptor (AChR) has been demonstrated in nonneurocrest-derived malignancies such as colon, prostate, lung, and ovarian carcinomas. Hence, targeted regulation of M3 AChR may be a possible mechanism for treating tumors. Aclidinium bromide has anti-tumoral properties in several tumors, namely gastric cancer and glioma. In this study, we intended to investigate whether aclidinium bromide, a novel M3 AChR antagonist, had effects on osteosarcoma cells proliferation and migration. PATIENTS AND METHODS: The viability of U2 OS cells was detected by cell counting kit-8 (CCK-8) assay. The migration and invasion capabilities were measured by transwell invasion and migration assays. The cell apoptosis rate was tested by Annexin V-fluorescein isothiocyanate (FITC)/Propidum iodide (PI) staining and flow cytometry. Key apoptosis-related and phosphatidylinositol-3-kinase (PI3K)/protein kinase B (AKT) signaling pathway-associated were assessed by Western blot analysis. RESULTS: Aclidinium bromide markedly decreased the OD value of U2 OS cells 48 h and 72 h after treatment. The number of positive crystal violet staining cells significantly decreased after treatment with aclidinium bromide. Treatment with aclidinium bromide significantly increased cell apoptosis rate, accompanied by the expression of anti-apoptotic protein Bcl-2 decreased, the expression of pro-apoptotic protein Active caspase-3 and Bax significantly increased in U2 OS cells treated with aclidinium bromide. Additionally, aclidinium bromide suppressed the PI3K/AKT signaling pathway in U2 OS cells. CONCLUSIONS: Therefore, the current study reveals that aclidinium bromide might inhibit osteosarcoma cell growth by regulating the PI3K/AKT signaling pathway, which suggests aclidinium bromide is a potential chemotherapeutic agent for osteosarcoma.

[Direct anterior approach versus posterolateral approach for total hip arthroplasty in the lateral decubitus position].

Objective: To compare the clinical efficacy of the direct anterior approach (DAA) and the posterolateral approach (PLA) for total hip arthroplasty (THA) in the lateral decubitus position. Methods: From July to December, 2014, 104 patients randomly divided into two equal groups of the DAA group and the PLA group underwent unilateral primary THA procedures.All procedures were performed by the same surgeon in the Department of Orthopaedics, the Affiliated Anhui Provincial Hospital of Anhui Medical University.General data, perioperative index, postoperative function and radiological evaluation were recorded and statistically analyzed. Results: The patients had an average follow-up of 14 (range, 10-16) months.No significant differences were detected with respect to the operation time, incision length, preoperative Hb concentration and transfusion rate between two groups (P> 0.05 for all comparisons). However, there were significant differences associated with the intraoperative bleeding, postoperative drainage and postoperative Hb concentration in the two groups(P<0.05 for all comparisons). The DAA group showed significant superior outcomes compared with the PLA group in the Harris hip scores [(83.6±7.1) vs (79.8±6.6), P<0.05], WOMAC[ (28.9±6.1) vs (36.1±6.9), P<0.001], and VAS pain scores[ (2.2±0.9) vs (2.9±1.1), P<0.05]at 1 month after surgery. No differences were seen between the study groups in the evaluation of radiography and the incidence of adverse event (P> 0.05 for all comparisons). Conclusions: Compared with the posterolateral approach, the present study shows the exciting results in patients underwent the DAA THA in the lateral decubitus position at early follow-up.The advantages of THA using the DAA include less operative trauma, alleviation of postoperative pain, and faster postoperative rehabilitation. It is a safe, reliable and effective surgery approach.

Aberrant DNA methylation of G-protein-coupled bile acid receptor Gpbar1 predicts prognosis of acute-on-chronic hepatitis B liver failure.

The G-protein-coupled bile acid receptor Gpbar1 (TGR5) has been demonstrated to be able to negatively regulate hepatic inflammatory response. In this study, we aimed to determine the methylation status of TGR5 promoter in patients with acute-on-chronic hepatitis B liver failure (ACHBLF) and its predictive value for prognosis. We enrolled 76 consecutive ACHBLF patients, 80 chronic hepatitis B (CHB) patients and 30 healthy controls (HCs). Methylation status of TGR5 promoter in peripheral mononuclear cell (PBMC) was detected by methylation-specific polymerase chain reaction (MSP). The mRNA level of TGR5 was determined by quantitative real-time polymerase chain reaction (RT-qPCR). We found that the frequency of TGR5 promoter methylation was significantly higher in ACHBLF (35/76, 46.05%) than CHB patients (5/80, 6.25%; χ(2) = 32.38, P < 0.01) and HCs (1/30, 3.33%; χ(2) = 17.50, P < 0.01). TGR5 mRNA level was significantly lower (Z = -9.12, P < 0.01) in participants with aberrant methylation than those without. TGR5 methylation showed a sensitivity of 46.05% (35/76), specificity of 93.75% (75/80), positive predictive value (PPV) of 87.5% (35/40) and negative predictive value (NPV) of 64.66% (75/116) in discriminating ACHBLF from CHB patients. ACHBLF patients with methylated TGR5 showed significantly poor survival than those without (P < 0.01). When used to predict 3-month mortality of ACHBLF, TGR5 methylation [area under the receiver operating characteristic curve (AUC) = 0.75] performed significantly better than model for end-stage liver diseases (MELD) score (AUC = 0.65; P < 0.05). Therefore, our study demonstrated that aberrant TGR5 promoter methylation occurred in ACHBLF and might be a potential prognostic marker for the disease.

Engineering of Yarrowia lipolytica lipase Lip8p by circular permutation to alter substrate and temperature characteristics.

Applications of lipases are mainly based on their catalytic efficiency and substrate specificity. In this study, circular permutation (CP), an unconventional protein engineering technique, was employed to acquire active mutants of Yarrowia lipolytica lipase Lip8p. A total of 21 mutant lipases exhibited significant shifts in substrate specificity. Cp128, the most active enzyme mutant, showed higher catalytic activity (14.5-fold) and higher affinity (4.6-fold) (decreased K m) to p-nitrophenyl-myristate (pNP-C14) than wild type (WT). Based on the three-dimensional (3D) structure model of the Lip8p, we found that most of the functional mutation occurred in the surface-exposed loop region in close proximity to the lid domain (S112-F122), which implies the steric effect of the lid on lipase activity and substrate specificity. The temperature properties of Cp128 were also investigated. In contrast to the optimal temperature of 45 °C for the WT enzyme, Cp128 exhibited the maximal activity at 37 °C. But it is noteworthy that there is no change in thermostability.

[The bioavailability of transdermal therapeutic system of timolol].

A matrix-type transdermal therapeutic system of timolol (TTS-timolol) was well prepared. The patch consisted of backing membrane layer, timolol reservoir layer, pressure sensitive adhesive layer and protective layer. A sensitive and reliable HPLC-UV method for the determination of plasma level of timolol in healthy volunteers was developed. Effective therapeutic plasma level of timolol (4 ng/ml) was attained 4 h after application of the timolol patches and was maintained within 32 h while the patch was removed at 24 h. The pharmacokinetic behavior of this transdermal therapeutic system (TTS)-timolol in human showed zero order absorption and well fitted to a one compartment model. The pharmacokinetic parameters are: Tmax = 18.8 h; Cmax = 11.2 ng/ml; AUC = 265.7 ng/ml.h; Vss = 120.0 L; K = 0.084 h-1. In comparison with the results of oral administration of timolol tablets, TTS-timolol possesses some advantages: stable plasma level, long effective time and convenient administration.