Objective To study the expression of active efflux pump AdeABC in clinical Acineto-bacter baumannii islates and whether this efflux pump confers resistance to antibiotics. Methods The anti-biotic susceptibility and the function of efflux pump inhibitor were tested by micro-dilution broth method. The expression of adeB was examined by RT-PCR. The controlling gene adeRS was amplified by PCR and se-quenced. Results Thirty multidrug resistance Acinetobacter baumannii isolates and 5 sensitive isolates for PCR were both abtained the expected products of adeB and adeRS. The mRNA expression of adeB in 15 multidrug resistance(MDR) isolates were positive, but there was no expression of adeB in 5 sensitive iso-lates. The mutations of adeRS existed in 2 MDR isolates. Conclusion The expression of AdeABC may in-volved in the resistant mechanisms of the clinical MDR Acinetobacter baumannii isolates.

AIM: To evaluate the safety and efficacy of cefetamet pivoxil vs cefixime in the treatment of acute bacterial infections. 　METHODS: A multicenter randomized controlled clinical trial was conducted. Ninety-eight patients (M 43, F 55; age 40 a±　s 13 a) with acute bacterial infections of cefetamet group were given cefetamet pivoxil 250-500 mg, po, bid, for 7-14 d, and ninty-five patients (M 44, F 51; age 42 a±14 a) of cefixime group were given cefixime 200 mg, po, bid, for 7-14 d.　RESULTS: The overall clinical efficacy rates were 95 % and 94 %, the bacterial clearance rates were 96 % and 94 %, the bacterial sensitive rates were 98 % and 96 %, the adverse reaction rates were 7 % and 6 %, respectively. There was no statistical significance between two groups (P>0.05).　CONCLUSION: Cefetamet pivoxil and cefixime are effective and safe in the treatment of acute bacterial infections.

Objective: To evaluate the clinical efficacy and safety of domestic faropenem in the treatment of acute bacterial infections. Methods: A multicenter, randomized, double blind and double simulation clinical study was conducted to compare the efficacy and safety of faropenem and cefaclor in the treatment of acute bacterial infection. Patients in trial group(n = 122) were given faropenem 250 mg，and in control group (n = 118) were given cefaclor 200 mg，3 times daily for 7 to 10 days.Results: The clinical cure rates were 33.61% and 27.12% in trail and control groups respectively and the clinical effective rates were 87.70% and 83.05% respectively. There was no significant difference in terms of clinical effectiveness between the two groups（P > 0.05）. The adverse reaction rates were 7.32% in trial group and 3.36% in control group（P > 0.05）. The adverse reaction of the trial group was mainly exaltation of aminotransferase, which did not affect the therapy. No severe adverse reaction was found.Conclusion: Domestic faropenem is effective and safe for the treatment of bacterial respiratory tract and urinary tract infections.

Objective To study the expression of active efflux pump AdeABC,AdeIJK,AdeFGH,AbeM,AbeS,CraA,MdtL in clinical Acinetobacter baumannii isolates and whether the efflux pumps confers resistance to antibiotics.Methods Thirty-two multi-drug resistant Acinetobacter baumannii islates and 10 sensitive isolates were collected.Genes of the exporter protein were amplified by PCR.The expression of adeB,adeJ,adeG,abeM,abeS,craA,mdtL were examined by real-time fluorescence quantitative RT-PCR.The controlling genes adeRS and adeL were amplified by PCR and sequenced.Results The positivity rates of adeB,adeJ,adeG,abeM,abeS,craA,mdtL were 100％,100％,100％,96.88％,100％,100％ and 93.75％ respectively in 32 multi-drug resistant Acinetobacter baumannii isolates,and all 100％ in 10 sensitive isolates.The difference of expression of adeB,abeM and mdtL were significant( P＜0.001,P =0.001,P=0.013) between 21 multi-drug resistant isolates of C clone and 10 sensitive isolates.The mutations of adeRS existed in 2 multi-drug resistant isolates,no point mutation of adeL.Conclusion The expression of AdeABC,AbeM and MdtL may involved in the resistant mechanisms of the clinical Acinetobacter baumannii islates.

Background and Objectives: This study was to investigate the role of microRNA-29a-3p (miR-29a-3p) in human bone marrow mesenchymal stem cells (hBMSCs), and its relationship with steroid-associated osteonecrosis. Methods: and Results: The online tool GEO2R was used to screen out the differentially expressed genes (DEGs) in GSE123568 dataset. Quantitative real time-polymerase chain reaction (qRT-PCR) was performed to detect the expression of miR-29a-3p, forkhead box O3 (FOXO3), alkaline phosphatase (ALP), bone gamma-carboxyglutamate protein (OCN) and RUNX family transcription factor 2 (Runx2) in the hBMSCs isolated from the patients with steroid-associated osteonecrosis. CCK-8 assay was executed to measure cell viability; western blot assay was utilized to detect FOXO3, ALP, Runx2, OCN and β-catenin expression. Cell apoptosis and cell cycle were detected by flow cytometry. Immunofluorescence assay was used to detect the sub-cellular localization of β-catenin. Bioinformatics analysis and luciferase reporter gene assay were performed to confirm whether miR-29a-3p can combine with FOXO3 3’UTR. MiR-29a-3p was markedly up-regulated in the hBMSCs of patients with steroid-associated osteonecrosis, while FOXO3 mRNA was significantly down-regulated. Transfection of miR-29a-3p mimics significantly inhibited the hBMSCs’ proliferation, osteogenic differentiation markers’ expressions, including ALP, Runx2, OCN, and repressed the ALP activity, as well as promoted cell apoptosis and cell-cycle arrest. FOXO3 was identified as a target gene of miR-29a-3p, and miR-29a-3p can inhibit the expression of FOXO3 and β-catenin, and inhibition of miR-29a-3p promoted translocation of β-catenin to the nucleus. Conclusions MiR-29a-3p can modulate FOXO3 expression and Wnt/β-catenin signaling to inhibit viability and osteogenic differentiation of hBMSCs, thereby promoting the development of steroid-associated osteonecrosis.

Mycosin-1 protease (MycP1) is a serine protease anchored to the inner membrane of Mycobacterium tuberculosis, and is essential in virulence factor secretion through the ESX-1 type VII secretion system (T7SS). Bacterial physiology studies demonstrated that MycP1 plays a dual role in the regulation of ESX-1 secretion and virulence, primarily through cleavage of its secretion substrate EspB. MycP1 contains a putative N-terminal inhibitory propeptide and a catalytic triad of Asp-His-Ser, classic hallmarks of a subtilase family serine protease. The MycP1 propeptide was previously reported to be initially inactive and activated after prolonged incubation. In this study, we have determined crystal structures of MycP1 with (MycP1²⁴⁻⁴²²) and without (MycP1⁶³⁻⁴²²) the propeptide, and conducted EspB cleavage assays using the two proteins. Very high structural similarity was observed in the two crystal structures. Interestingly, protease assays demonstrated positive EspB cleavage for both proteins, indicating that the putative propeptide does not inhibit protease activity. Molecular dynamic simulations showed higher rigidity in regions guarding the entrance to the catalytic site in MycP1²⁴⁻⁴²² than in MycP1⁶³⁻⁴²², suggesting that the putative propeptide might contribute to the conformational stability of the active site cleft and surrounding regions.
Amino Acid Sequence ; Bacterial Proteins ; chemistry ; Bacterial Secretion Systems ; Crystallography, X-Ray ; Humans ; Molecular Dynamics Simulation ; Molecular Sequence Data ; Mycobacterium smegmatis ; metabolism ; Protein Precursors ; chemistry ; Protein Structure, Tertiary ; Subtilisins ; chemistry

Objective To understand the situations of departments of intensive care units (ICUs) of different level hospitals in Guizhou Province, and to provide directions and evidences for improving quality control in critical care medicine.Methods A county-level hospital and a provincial-level hospital's comprehensive ICU in Guizhou Province were selected to record and analyze and compare the structural indicators, patient admission and transfer, disease distribution, ventilator associated pneumonia (VAP), intravascular catheter related blood stream infection (CRBSI) and catheter-associated urinary tract infection (CAUTI) of the two hospitals' comprehensive ICU in 2017.Results The ICU of the People's Hospital of Suiyang County (county hospital) was found in 2012, and the ICU of the Affiliated Hospital of Guizhou Medical University (provincial hospital) was found in 1994. Until 2017, there were 10 and 46 beds, 6 (all of them hold bachelor's degree) and 18 physicians (6 of them hold PhD, 5 of them hold master's degree, 7 of them hold bachelor's degree), 17 (4 of them hold bachelor's degree, 13 of them hold college degree or graduated from secondary school) and 69 nurses (2 of them hold master's degree, 53 of them hold bachelor's degree, 14 of them hold college degree or graduated from secondary school) in the two ICUs respectively, there were significant differences in the education background of the physicians and nurses between the two ICUs (bothP < 0.01). During 2017, 471 cases were admitted to the ICU of the county hospital while 1633 cases were admitted to the ICU of the provincial hospital. Compared with the ICU of the provincial hospital, the ratio of the patients with acute physiology and chronic health evaluationⅡ(APACHE Ⅱ) ≥ 15 at admission was lower (74.8％ vs. 85.1％,P < 0.01), the ratio of direct admission was higher (30.8％ vs. 17.4％,P < 0.01), the ratio of the patients admitted to the ICU more than once was lower (0.8％ vs. 5.0％,P < 0.01), the ratio of the patients whose the length of ICU stay less than 24 hours was higher (51.6％ vs. 13.7％,P < 0.01), the ratio of the patients whose the length of ICU stay more than 28 days was lower (1.1％ vs. 2.9％, P < 0.05), the ratio of the patients discharged against-advice (25.5％ vs. 20.5％,P < 0.05) was higher, the ratio of the patients transferred to other hospitals was higher (5.1％ vs. 0.3％,P < 0.05), and the ICU mortality was lower (4.0％ vs. 13.9％,P < 0.01) in the ICU of the county hospital. The top three kinds of diseases treated in the ICU of the county hospital were brain injury (27.4％), trauma (19.1％) and toxication (6.8％); while in the ICU of the provincial hospital were brain injury (18.6％), sepsis (16.2％) and severe acute pancreatitis (4.8％). In addition, the incidences of VAP, CRBSI and CAUTI in the ICU of the county hospital were 10.0/1000 ventilator days, 1.4/1000 catheter days, 0.5/1000 catheter days; while in the ICU of the provincial hospital were 5.8/1000 ventilator days, 2.0/1000 catheter days, 3.7/1000 catheter days, respectively.Conclusions There are short of physicians and nurses in the ICU of the provincial and county hospitals in Guizhou Province, and the educational level of the medical staff in the ICU of the county hospital is relatively low. Moreover, there were significant differences in the admissions and treatments and the outcomes of the critically ill patients between the two ICUs. The characteristics of the ICUs of county hospitals should be fully considered when the quality control of critical care medicine and continuing medical education are done.

Pleomorphic adenoma (PA) is the most common benign tumour in the salivary gland and has high morphological complexity. However, the origin and intratumoral heterogeneity of PA are largely unknown. Here, we constructed a comprehensive atlas of PA at single-cell resolution and showed that PA exhibited five tumour subpopulations, three recapitulating the epithelial states of the normal parotid gland, and two PA-specific epithelial cell (PASE) populations unique to tumours. Then, six subgroups of PASE cells were identified, which varied in epithelium, bone, immune, metabolism, stemness and cell cycle signatures. Moreover, we revealed that CD36⁺ myoepithelial cells were the tumour-initiating cells (TICs) in PA, and were dominated by the PI3K-AKT pathway. Targeting the PI3K-AKT pathway significantly inhibited CD36⁺ myoepithelial cell-derived tumour spheres and the growth of PA organoids. Our results provide new insights into the diversity and origin of PA, offering an important clinical implication for targeting the PI3K-AKT signalling pathway in PA treatment.
Humans ; Adenoma, Pleomorphic/genetics* ; Phosphatidylinositol 3-Kinases ; Proto-Oncogene Proteins c-akt ; Transcriptome ; Myoepithelioma