OBJECTIVEThis study aims to explore the effect of Notch signaling depression on the receptor activator of nuclear factor kappa B ligand (RANKL)-induced osteoclastogenesis of RAW264.7 cells.

METHODSMice RAW264.7 cells were cultured and differentiated into osteoclasts with the induction of RANKL. The expressions of Notch1, Notch2, Deltal, Jagged1, Hes1, tartrate-resistant acid phosphatase (TRAP), and Cathepsin K genes during osteoclastogenesis were analyzed using real-time polymerase chain reaction. Osteoclast formation was analyzed using TRAP assay with suppression of Notch receptors by a selective γ-secretase inhibitor (GSI).

RESULTSNotch1, Notch2, Delta1, Jagged1, and Hes1 expressions in RAW264.7 cells were upregulated following 50 ng · mL-RANKL stimulation for 3 d, concomitant with the expression of the osteoclast differentiation markers TRAP and Cathepsin K. Notch2 and Jagged1 had the most remarkable increase in the Notch family members. GSI inhibited RANKL-induced osteoclastogenesis of RAW264.7 cells and Hes1 expression dose-dependently.

CONCLUSIONNotch signaling activation may promote RANKL-induced osteoclastogenesis of RAW264.7 cells.

Objective To evaluate the appropriateness of hospitalization days at a tertiary hospital in 2014 by means of the Appropriateness Evaluation Protocol ( AEP ) , and to analyze the causes of inappropriate stays. Methods Medical records of inpatients admitted at a tertiary hospital in 2014 were randomly selected. AEP( US version) was used to evaluate the appropriateness of every hospitalization day, while the causes of inappropriate hospitalization day were also analyzed. Results A total of 1 641 days of stay from 148 medical records were reviewed, and 129 days of stay (7. 9%) were seen as inappropriate. Two major factors for inappropriate stays were waiting for surgery and waiting for test, roughly 89. 1% of the inappropriate hospitalization days. The proportion of inappropriate hospital stays reduced to 4. 8% after adjustment of two-day weekend. Inappropriate hospital stays mostly appeared during the second day to the eighth day after admission(93. 8%). Logistic analysis results showed that with concomitant symptoms, preoperative waiting days > 5 days, high level surgery, non-emergency admission were significantly associated with appropriateness of hospital stays (P<0. 05). Conclusions The rate of inappropriate stays will be reduced and the quality of medical services will be improved if comprehensive measures could be carried out according to the causes of inappropriate stays.

Objective To develop and validate a ultrasonographic (US)imaging agent with targeted microbubbles that attaches to chemokine receptor 2 (CCR2)and to compare the US single obtained from targeted microbubble with that from control microbubble in murine breast tumor model.Methods The microbubble which carried CCR2 antibody (MBCCR2 )and isotype-macthed immunoglobulin G-labled control microbubble (MBcontrol ) were prepared. The microbubble size and distribution were assessed by AccuSizer780.Binding specificities of targeted microbubble compared with control microbubble were tested with murine microvascular endothelial cells (bEnd.3 ).Orthotopic breast tumor model was estabished in BALB/c mice with mouse breast cancer 4T1 cell.In vivo imaging signals of contrast material-enhanced ultrasound by use these two different types of microbubble which were injected respectively into each mouse at random order and 30 min interval.Tumor tissue was stained for CCR2 and CD3 1 .Results Automatic Particle Sizer showed size uniform of two kinds of microbubbles,and narrow distribution of particle size (mean diameter of about 1 -2 μm),which were not significantly different (P >0.05).Adhension to bEnd. 3 endothelial cells was significantly higher (P < 0.001 )for MBCCR2 (mean,9.50 ± 1 .5 1 )than that for MBcontrol (mean,0.01 ±0.01).Imaging signal in the murine tumor model was significantly higher for MBCCR2 [mean,(6.76±0.26)dB]than that for MBcontrol [mean,(1 .06 ±0.62)dB,P <0.001 ].Immunofluorescence confirmed expression of CCR2 on tumor vasculature.Conclusions The targeted microbubbles with CCR2 monoclonal antibody had been successfully prepared,which precisely targeted to CCR2 of tumor angiogenesis in the murine breast cancer xenograft tumor models in vivo.These results suggest that the targeted microbubbles as a kind of ultrasound molecular imaging agent with a better specificity can be used for both evaluating tumor neovascularization and monitoring therapeutic effect of anti-angiogenesis.

Objective To investigate the present status and developments of Chinese military neurosurgery, make an objective analysis about the achievements and deficiencies, and propose a developing trend for the field of Chinese military neurosurgery. Methods The scientific literatures of neurosurgery published domestically and abroad in recent 5 years were partially reviewed. The advances and achievements in basic researches and clinical works of neurosurgery acquired during the period of the 11th Five Year Plan were analyzed. Results During the period of the 11th Five Year Plan, Chinese military neurosurgery was still in the lead position in traditional areas such as traumatic brain injury. In the meantime, great achievements were obtained in the researches of brain and spinal cord tumors, endovascular treatment and functional neurosurgery, and 47 items of Army Science and Technology Advancement Prizes or Military Medical Achievement Awards were acquired. A great progress was made in personnel training. However, unbalanced development in neurosurgery field still existed within military departments, between military and civil departments and between different regions, so an enhanced technical interchange and cooperation is needed. Conclusions On the coming period, the research work of military neurosurgery should focuse on basic and clinical research of gene therapy to keep in step with neurosurgical trends in the world. On the other hand, attention should be paid to technological innovation with military features such as professional and general treatment of traumatic brain injury and researches on the development of protective equipments.

Aged ; Female ; Glossopharyngeal Nerve Diseases ; surgery ; Humans ; Light Coagulation ; Lingual Nerve ; surgery ; Male ; Microwaves ; therapeutic use ; Middle Aged

Objective To investigate the feasibility of recombinant adeno-associated virus encoding secrected forms of human acidic fibroblast growth factor transfecting endothelial progenitor cells.Methods sp-haFGF was obtained through combining signal peptide sequence of FGF-4 with native aFGF gene by PCR.sp-haFGF was cloned into AAV vector plasmid pAAV-IRES-hrGFP.Recombinant AAV encoding sp-haFGF was packaged through co-transfecting HEK293 cells with plasmid sp-haFGF-pAAV-IRES-hrGFP,pAAV-RC and pHelper.Ex vivo cultured EPCs were infected with concentrated rAAV.Expression of green fluorescent protein(GFP) in infected EPC was observed by fluorescence microscope and expression of sp-haFGF in EPCs was verified by RT-PCR and western blot analysis.Results Recombinant AAV encoding sp-haFGF was obtained.After EPCs being infected with rAAV,green fluorescence was found in about 20～30% EPCs,gene of(560 bp) was generated from EPCs by RT-PCR method,and(sp-haFGF) protein was detected in infected cells.Conclusion EPC was efficiently infected by rAAV encoding(sp-haFGF) and(haFGF) was expressed by EPCs,which establishes a basis for therapeutic angiogenesis by transgenic EPCs transplantation.

The readjustment of the army establishments has resulted in the mergence of some sanatoriums into hospitals,which has given rise to a novel mode of management and offered a new opportunity for the development of sanatoriums.In the past four years since the mergence of our sanatorium into Nanjing General Hospital in 2004,we explored the mode of integrating recuperation with treatment and combined the resource advantages of ours with the technical superiority of the hospital,which led to an improvement of our competitive ability and comprehensive service and accordingly promoted the overall development of the sanatorium.

Objective To explore optimal initial the best screening time for newborns with different delivery methods using AABR.Methods A total of 550 newborns who were born from August 1, 2016 to October 31, 2016 at our hospital participated in the study.AABR was used to accomplish the initial hearing screening.The newborns were divided into 2 groups according to the delivery methods.There were each 100 neonates born in vaginal during <24 h, 24~48 h and 48~72 h after birth, respectively.The numbers of neonates delivered by cesarean section during the 3 separate periods were 50, 100 and 100, respectively.The newborns who failed the preliminary hearing screening proceeded to the re-screening and diagnostic procedures.Results There were 300 newborns were born in vaginal, and the pass rate in 24~48 h after birth group was significantly higher than that in 24 h group (93.00% vs 83.00%,x2=4.735,P=0.03<0.05), but it was not significantly different from that of 48~72 h group (95.00% vs 93.00%,x2=0.355,P=0.56>0.05).There were 250 newborns in cesarean section, the pass rate of 24~48 h after birth group was significantly higher than that in 24 h group (83.00% vs 68.00%,x2=4.437, P=0.04<0.05), and significantly lower than that of 48~72 h group (94.00% vs 83.00%,x2=5.944, P=0.02<0.05).Conclusion Taking into account of hospitalization time, the screening time for the vaginal delivery newborn hearing screening can be advanced to 24~48 h after birth with the application of AABR, but not for the cesarean section group.

Background Retinal neovascularization varies with the change of microRNA (miRNA) expression level.Quantitative real-time PCR (qRT-PCR) is a common method for the analysis of miRNA expression profiling.However,the housekeeping genes selected as references may exhibit differential expression levels under the distinctive experimental conditions.The accuracy of the levels of target gene expression often is affected if the selected housekeeping genes with significantly fluctuating expression as references.Determining a reference gene with stable expression level is the premise to consecutive studies.Objective This study was to compare the expression levels and stability of the frequently used reference genes for miRNA expression analysis in normally developing eyes and in eyes of a mouse model of oxygen-induced retinopathy (OIR),and select the optimal reference gene (s) exhibiting stable ocular expression under both hypoxia and normal development conditions.Methods P7,P12,P17,P37 and 8-week-old clean C57BL/6J mice were selected randomly.q-PCR was used to detect the dynamic changes of relative expressions of 5 kinds of reference genes in different ages of mice,including snRNAU6 (RNU6),5S ribosomal RNA (5s rRNA),snoRNA U68 (U68),snoRNA U70 (U70),snoRNA U49A (U49A).Other 40 P7 mice were randomized into the normal control group and the OIR group.The mice of the normal control group were fed in the normal oxygen environment,and those in the OIR group were raised in the (75-±2)％ oxygen environment for 10 days.Fluorescine was injected via ritrobulbar vein for retinal stretched preparation,and the histopathological examination of mouse retinas was performed to identified the models.The expressing difference of 5 kinds of reference genes in the retinas were detected to compare the differences of 5 kinds of genes expression between the two groups.GeNorm program was employed to compare the stability of the expressing genes.This study were approved and granted by Ethical Committee of Tianjin Medical University and met the requirements stipulated in the Association for Research in Vision and Ophthalmology Statement for the Use of Animals in Ophthalmic and Vision Research.Results Across the developmental ages,the expression levels of U68 in the eye exhibited the greatest variability,and then coming with U49A,U70,RNU6 and 5s rRNA.Significant differences were seen across the developmental ages for the expression levels of U68,U49A,U70,and RNU6 (U70:F =7.005,P =0.000 ; U68:F =10.189,P =0.000 ; U49A:F =21.134,P=0.000;RNU6:F=4.968,P=0.004).Expression of 5s rRNA showed the least variability across the developmental ages (F=2.099,P =0.107).In P17 mice of the OIR group,tortuous access vessels and non-perfusion area were found in the retinal section.Hematoxylin-eosin stain showed the neovascular sprout through across the inner limiting membrane.The relative expression of U70 exhibited the greatest variability and then were U49A,U68 and 5s rRNA in turn in P17 OIR mice,and significantly elevated expressions were found in U70,U49A,U68 and 5s rRNA genes between the OIR group and the normal control group (t =5.174,P =0.000;t =3.376,P =0.012;t =4.802,P =0.000 ;t=2.856,P=0.029).Expression of RNU6 showed the least variability between the two groups (t =2.104,P=0.065).As analyzed by GeNorm program,the stability for the five reference genes across developmental ages was 5s rRNA/RNU6＞ U70 ＞ U49A ＞ U68,and the optimal reference combination was 5s rRNA and RNU6.Whereas the stability for the OIR model was 5s rRNA/RNU6＞U68＞ U49A＞U70,and the optimal reference combination was 5s rRNA and RNU6.Conclusions Reference genes should be selected based on specific subjects and experimental conditions when qRT-PCR is used to analyze miRNA expression levels.In the OIR model,both developmental and hypoxic factors need to be considered.5s rRNA and RNU6 reference combination is the preferred option for the qRT-PCR analysis of ocular miRNA expression if available.

In this paper, the process performance indexes (PPIs) P(p) and P(pk) were introduced and applied to evaluate the process capability and quality consistence of Chinese medicine products. The historical quality analysis data of Qingkailing injection were collected and taken as the research object. The confidence intervals of P(p) and P(pk) were estimated based on the Bootstrap sampling methods. Results showed that the value and width of the confidence interval P(p) of were smaller that of P(pk) suggesting that the P(pk) index was more sensitive than P(p) in process capability analysis. Within the production period concerned, the P(pk) values estimated from different quality indexes of Qingkailing injection, such as baicalin, cholic acid, geniposide and total nitrogen, were 1.122, 2.055, 1.564 and 0.891, respectively. It could be found that the cholic acid had the highest process capability, followed by the geniposide and baicalin. The total nitrogen had the lowest process capability, indicating that it is necessary to reinforce the quality management of total nitrogen related manufacturing processes. The case studies demonstrate the effectiveness and feasibility of PPIs, which are convenient to be used in production practice of Chinese medicine.
Chemistry, Pharmaceutical ; methods ; standards ; Drugs, Chinese Herbal ; chemistry ; standards ; Injections ; Quality Control