Objective To construct the recombinant plasmid of PA-ⅠL and express in E.coli BL 21 (DE3), and evaluate the biological activity of recombinant protein.Methods PA-ⅠL gene was amplified by PCR using primers designed according to Pseudomonas aeruginosa genome sequences and then cloned to the vector pET -28a ( +).The recombinant plasmid was transformed into E.coli BL21(DE3) and induced to express by IPTG.The recombinant protein was purified by nickel affinity chromatography.The binding activity of recombinant PA-ⅠL with Gb3/CD77 was evaluated by flow cytometry.The function of recombinant PA-ⅠL on the binding of bacteria with host cells was evaluated by colony plate counting.Results and Conclusion The recombinant PA-ⅠL protein was highly expressed in E.coli BL21(DE3) and protein purity by SDS-PAGE analysis was high after nickel affinity chromatography .Besides, the recombinant PA-ⅠL had binding activity to Gb3/CD77 and inhibited the binding of PAO1 to host cells in a dose-dependent manner.

Objective To evaluate the value of utility of bronchoscopy in human immunodeficiency virus/acquired immunodeficiency syndrome (HIV/AIDS) patients with Bacteria sputum negative pulmonary tuberculosis.Methods Bronchoscopy was conducted to 65 AIDS patients with bacteria sputum negative pulmonary tuberculosis in the first hospital of Changsha. The patients’ bronchoalveolar lavage fluid through the electronic bronchoscopy, mycobacterium tuberculosis (MTB) culture, brushings and biopsy pathology were analyzed.Results 65 cases, bronchoscope alveolar lavage lfuid smear positive acid-fast stain 14 cases (21.54%), BAL mycobacterium tuberculosis culture positive 20 cases (30.76%), a bronchoscope brush positive 24 cases (36.92%), 35 cases of bronchoscopy biopsy, according to the performance under the bronchoscope positive 21 cases (60.00%), bronchoscopy combined different methods conifrmed 43 cases (66.15%).Conclusions Bronchoscopy in AIDS with bacteria sputum negative pulmonary tuberculosis diagnosis, it has important application value.

Objective To evaluate the application and effects of mind map in nursing handover. Methods Two wards which had similar scale, environment, property and level of nursing were randomly chosen as the experimental group and the control group by tossing a coin in Tianjin Hospital of ITCWM Nankai Hospital, 28 nurses were selected in each group, according to the inclusion and exclusion criteria, control group takes routine handover, the experimental group takes handover by following the guidance of mind map. The mind map of clear mind were designned. The Chinese version of Handover Evaluation Scale was used to evaluate the effects of mind map in the nursing handover, the inpatients'satisfaction was compared between the experimental group and the control group after the application of min-d map. Results After3-month intervention, results showed that the score of handover evaluation scale in the experimental group were (31.39 ± 1.34) points, (19.57 ± 1.23) points, (17.61 ± 1.29) points, (68.57 ± 2.49) point, the score of handover evaluation scale in the control group were (26.71±1.24) points,(18.71±1.33) points,(16.68 ± 1.33) points, (62.11 ± 2.94) points, the difference was statistically significant(t=2.504-13.530, P<0.05);in the experimental group, the score of handover evaluation scale increased significantly than that before intervention, and the difference was statistically significant (t=-17.450--5.663, P<0.05);the inpatients′satisfaction in the experimental group was 98.89%(89/90), the control group was 90%(81/90) after intervention, and the difference was statistically significant(χ2=6.776, P<0.05); in the experimental group, the inpatients′satisfaction was better than that before intervention, and the difference was statistically significant (χ2=4.211, P<0.05) .Conclusion:The application of mind map in the n-ursing handover can improve the level of nursing handover and the quality of nursing care, contributing to dveloping high quality of nursing service.

OBJECTIVE: To evaluate the auditory performance of infants with cochlear implants at the early stage after surgery, summarize the development of auditory ability, investigate the effect of age at cochlear implantation on auditory performance within the first year after surgery and provide a reference for their habilitation program. METHOD: A total of 272 infants with prelingually profound hearing loss participated in this study, ranging in age at cochlear implantation from 18 to 36 months. The mean age was 21 months with a standard deviation of 7 months. Infants with cochlear implants were divided into three groups according to their age at implantation. Infants in group A were implanted under 18 months of age. Infants in group B were implanted between 18 and 24 months of age. Infants in group C were implanted between 25 and 36 months of age. Categories of auditory performance (CAP) was used to evaluate the auditory performance, which rates auditory abilities in eight categories for a scale of 0 to 7. RESULT: The mean scores of CAP for all infants at each interval were significantly different after implantation. Significant differences were observed in mean scores of CAP among these three groups in 1 and 3 months after switch-on. However there were no significant differences in pre-operation, 6, 9 and 12 months after switch-on. CONCLUSION There is a significant improvement in auditory performance of infants with prelingually profound hearing loss within the first year after cochlear implantation. The age at cochlear implantation has no critical influence on the development of auditory capabilities at the early stage after surgery. CAP is a practical tool which can be used in clinic in China.
Age Factors ; Child, Preschool ; Cochlear Implantation ; Cochlear Implants ; Deafness ; surgery ; Female ; Hearing ; Humans ; Infant

Objective: To investigate the outbreak of mumps in a remote village school of Zhaoqing City, to evaluate and vaccine effectiveness ( VE ) of mumps containing vaccine (MuCV), so as to provide reference for the formulation of epidemic prevention and control strategies. Methods: Through on site case investigations and interviews, case data and epidemic related epidemiological information were obtained. Descriptive and retrospective cohort studies were used to analyze the epidemic characteristics, explore risk factors, and evaluate the protective effect of vaccines. Results: Totally 166 cases of mumps were found, and all of them were students. The total attack rate was 7.79% and the outbreak lasted for 60 days. The first case occurred on October 11th and the last case occurred on December 9th in 2018. Most of the cases aged from 10 to 13 years old, accounting for 66.27%. There were 96 male patients and 70 female patients, with no statistically significant sex difference in the incidence rate ( χ 2=2.40, P >0.05). Involving 28 classes, 11 of which had an incidence rate more than 10%, mainly distributed in grades 2, 4 and 6. There were statistically significant differences in incidence rates among different grades ( χ 2=96.89, P <0.01) and different floors ( χ 2=67.35, P < 0.01 ), with the third floor higher than the other floors. Twelve out of 58 boarding pupils were cases, and boarding pupils were 1.89 times higher in risk of contracting mumps than day students ( RR=1.89, 95%CI =1.10-3.23). Pupils without being given the shot of MuCV were higher in the infection rate than those having the shot ( χ 2=5.70, P <0.05), and the VE % was 35% (95% CI = 7%- 55%). The VE % of one dose was 34% (4%-54%), while the effectiveness of protection was declined with time ( χ 2 trend =6.53, P < 0.05). The effectiveness of vaccine almost diasappeared six years after the shot ( χ 2=1.12, P >0.05). Conclusion Delayed case report and isolation, low rate of receiving MuCV, and decreasing effectiveness of one shot MuCV are closely assocaited with the outbreak and ongoing spread of the epidemic.

Clinical residual biological specimens are invaluable for medical research and can be reused for medical research. This paper expounded the possibility, necessity and applied range of the medical research reuse of clinical residual biological specimens and put forward some suggestions on how to standardly supervise clinical residual biological specimens for medical research reuse. The authors raised four aspects of concern: how to strengthen the management of ethical review; how to establish strict privacy protection and information confidentiality system; how to keep samples reasonably to ensure clinical examination; and how to ensure the compliance treatment of residual biological specimens after reuse, so as to promote clinical residual biological specimens more normatively and effectively used in medical research.

Chronic myeloid leukemia (CML) is a slowly progressing hematopoietic cell disorder. Sphingosine kinase 1 (SPHK1) plays established roles in tumor initiation, progression, and chemotherapy resistance in a wide range of cancers, including leukemia.However, small-molecule inhibitors targeting SPHK1 in CML still need to be developed. This study revealed the role of SPHK1 in CML and investigated the potential anti-leukemic activity of hirsuteine (HST), an indole alkaloid obtained from the oriental plant Uncaria rhynchophylla, in CML cells. These results suggest that SPHK1 is highly expressed in CML cells and that overexpression of SPHK1 represents poor clinical outcomes in CML patients. HST exposure led to G2/M phase arrest, cellular apoptosis, and downregulation of Cyclin B1 and CDC2 and cleavage of Caspase 3 and PARP in CML cells. HST shifted sphingolipid rheostat from sphingosine 1-phosphate (S1P) towards the ceramide coupled with a marked inhibition of SPHK1. Mechanistically, HST significantly blocked SPHK1/S1P/S1PR1 and BCR-ABL/PI3K/Akt pathways. In addition, HST can be docked with residues of SPHK1 and shifts the SPHK1 melting curve, indicating the potential protein-ligand interactions between SPHK1 and HST in both CML cells. SPHK1 overexpression impaired apoptosis and proliferation of CML cells induced by HST alone. These results suggest that HST, which may serve as a novel and specific SPHK1 inhibitor, exerts anti-leukemic activity by inhibiting the SPHK1/S1P/ S1PR1 and BCR-ABL/PI3K/Akt pathways in CML cells, thus conferring HST as a promising anti-leukemic drug for CML therapy in the future.

BACKGROUNDIn-stent restenosis caused by airway granulation poses a challenge due to the high incidence of recurrence after treatment. Weekly applications of anti-proliferative drugs have potential value in delaying the recurrence of airway obstruction. However, it is not practical to subject patients to repeated bronchoscopy and topical drug applications. We fabricated novel pacilitaxel-eluting tracheal stents with sustained and slow pacilitaxel release, which could inhibit the formation of granulation tissue. And we assessed the quality and drug release behaviors of drug-eluting stents (DESs) in vitro.

METHODSStents were dipped vertically into a coating solution prepared by dissolving 0.5 g (2% w/v) of poly lactic acid-coglycolic acid (PLGA) and 0.025 g (0.1% w/v) of pacilitaxel in 25 ml of dichloromethane. DES morphology was examined by scanning electron microscopy (SEM). Pacilitaxel release kinetics from these DESs was investigated in vitro by shaking in PBS buffer followed by high performance liquid chromatography (HPLC).

RESULTSUsing an orthogonal experimental design, we fabricated numerous pacilitaxel/PLGA eluting tracheal stents to assess optimum coating proportions. The optimum coating proportion was 0.1% (w/v) pacilitaxel and 2% (w/v) PLGA, which resulted in total pacilitaxel loading of (16.380 6 ± 0.002 1) mg/stent. By SEM the coating was very smooth and uniform. Pacilitaxel released from DES was at (0.376 3 ± 0.003 8) mg/d, which is a therapeutic level. There was a prolonged, sustained release of pacilitaxel of >40 days.

CONCLUSIONSPaclitaxel-loaded PLGA coating tracheal stents were successfully developed and evaluated. Quality assessments demonstrated favorable surface morphology as well as sustained and effective drug release behavior, which provides an experimental reference for clinical practitioners.

Drug-Eluting Stents ; Glycolates ; chemistry ; Humans ; Lactic Acid ; chemistry ; Paclitaxel ; chemistry ; Polyesters ; Polymers ; chemistry

The phosphatidylinositol 3-kinase (PI3K) pathway is frequently activated in human cancers. Class I PI3Ks are lipid kinases that phosphorylate phosphatidylinositol 4,5-bisphosphate (PIP2) at the 3-OH of the inositol ring to generate phosphatidylinositol 3,4,5-trisphosphate (PIP3), which in turn activates Akt and the downstream effectors like mammalian target of rapamycin (mTOR) to play key roles in carcinogenesis. Therefore, PI3K has become an important anticancer drug target, and currently there is very high interest in the pharmaceutical development of PI3K inhibitors. Idelalisib has been approved in USA and Europe as the first-in-class PI3K inhibitor for cancer therapy. Dozens of other PI3K inhibitors including BKM120 and ZSTK474 are being evaluated in clinical trials. Multifaceted studies on these PI3K inhibitors are being performed, such as single and combinational efficacy, resistance, biomarkers, etc. This review provides an introduction to PI3K and summarizes key advances in the development of PI3K inhibitors.

RBM46 is a germ cell-specific RNA-binding protein required for gametogenesis, but the targets and molecular functions of RBM46 remain unknown. Here, we demonstrate that RBM46 binds at specific motifs in the 3'UTRs of mRNAs encoding multiple meiotic cohesin subunits and show that RBM46 is required for normal synaptonemal complex formation during meiosis initiation. Using a recently reported, high-resolution technique known as LACE-seq and working with low-input cells, we profiled the targets of RBM46 at single-nucleotide resolution in leptotene and zygotene stage gametes. We found that RBM46 preferentially binds target mRNAs containing GCCUAU/GUUCGA motifs in their 3'UTRs regions. In Rbm46 knockout mice, the RBM46-target cohesin subunits displayed unaltered mRNA levels but had reduced translation, resulting in the failed assembly of axial elements, synapsis disruption, and meiotic arrest. Our study thus provides mechanistic insights into the molecular functions of RBM46 in gametogenesis and illustrates the power of LACE-seq for investigations of RNA-binding protein functions when working with low-abundance input materials.
Animals ; Mice ; 3' Untranslated Regions/genetics* ; Cell Cycle Proteins/metabolism* ; Gametogenesis/genetics* ; Meiosis/genetics* ; Nuclear Proteins/genetics* ; RNA-Binding Proteins/genetics*