AIM: To study the changes of bacterial ultrastructure, membrance potential and membrance patency of Escherichia coli during the postantibiotic effect after exposure to gatifloxacin and ciprofloxacin in order to investigate the mechanisms of PAE. METHODS: During the Postantibiotic effect after exposure to gatifloxacin and ciprofloxacin, the aliquots were taken from the bacterial culture at regular intervals. Then the fluorescence microscope was used to observe changes in bacterial ultrastructure and at the same time we studied the changes of membrance potential and membrance patency in Escherichia coli during the postantibiotic effect by flow cytometry in conjunction with fluorescent probes. RESULTS: The PAE phase characterized by filament formation after exposure to gatifloxacin by fluorescence microscopy,yet no significant changes in membrance potential and membrance patency of Escherichia coli were observed. CONCLUSION: Gatifloxacin and ciprofloxacin can induce filamentation, and this change is indifferent with membrance potential and membrance patency of Escherichia coli.

OBJECTIVE:To provide reference for further improving negotiation mechanism of drug medical insurance in Chi-na. METHODS:Questionnaire investigation was conducted among the medical insurance agency and pharmaceutical company staff who participated in drug medical insurance negotiation in Jiangsu province,Jiangxi province,Qingdao,Chengdu. Their business fa-miliarity and recognition of the negotiating mechanism were also investigated. The descriptive analysis,independent sample t test and other methods were adopted to analyze the data statistically. RESULTS:A total of 70 questionnaires were distributed and 68 val-id questionnaires were collected with effective recovery rate of 97.14%. Respondents included 21 staff members of medical insur-ance agencies and 47 of pharmaceutical companies. In terms of business familiarity,the average score of respondents in all aspects was greater than 3(4 points system),the lowest score wasmanagement of the implementation of drug negotiation projects. There were statistical significances in scores of 3 items between 2 parties in the negotiation(P<0.05):basic principles of the drug negotiation process should be followednegotiation objectives in the substantive negotiation process total score of busi-ness familiarity. The scores of respondents in pharmaceutical companies were higher than those of health insurance agencies. In the recognition of the negotiating mechanism,average score of each survey item was ≥4(5 points system)in the recognition of the overall effect of the negotiation mechanism except forwhich is conducive to maintaining the price system of pharmaceutical com-panies. The average score of all aspects of preparation of the negotiations,substantive negotiation,implementation of the negotia-tion agreement and negotiation project's supervision and evaluation was >4 except forpatients taking drugs is quick and easy. There was no statistical significance between 2 types of respondents in all aspects of overall effect of the negotiation mechanism, preparation of the negotiations and negotiation project's supervision and evaluation(P>0.05). But in the recognition of the substan-tive negotiation,there was statistically significance between 2 types of respondents innegotiations between the two sides in an equal,cooperative,harmonious atmosphere(P<0.05). Healthcare agencies'respondents had higher scores than pharmaceutical companies'ones. And in the recognition of the implementation of the negotiation agreement,there was statistical significance be-tween 2 types of respondents in theafter the agreement entered into force,the medical insurance agency will be strictly in accor-dance with the agreementandfund pay to pharmaceutical companies timely and reasonable(P≤0.05). Healthcare agencies're-spondents have higher scores for than pharmaceutical companies'ones. CONCLUSIONS:The current drug medical insurance nego-tiation mechanism shows good overall effect and runs smoothly. But there are still some shortages,such as collected information the preparation stage is not sufficient;the procedures of patients taking the drug is complicated;negotiating parties do not have equal status;the fund is not paid to the pharmaceutical enterprise timely;evaluation is not perfect.

Background and purpose:Pterostilbene is a natural antioxidant, whose role in retinoblastoma remains unclear. The aim of this study is to probe the effects of pterostilbene on the proliferation, apoptosis and autophagy in retinoblastoma WERI-Rb-1 cell lines.Methods:Cell counting kit-8 (CCK-8) assays were used to analyze the effects of pterostilbene on the proliferation of WERI-Rb-1 cells. Apoptosis rate was determined by Annexin V/PI. Autophagic vacuoles were observed by acridine orange staining. LC3 and P62 protein expressions were determined using Western blot.Results:Pterostilbene significantly inhibited the proliferation of WERI-Rb-1 cells (P<0.01). The cell viability were (93.02±0.47)%, (55.10±2.04)% and (30.33±1.45)% after WERI-Rb-1 cells were treated with 25, 50 and 100 μmol/L pterostilbene for 24 h, and the cell viability were (88.38±3.70)%, (53.37±1.17)%, (29.60±1.05)% after WERI-Rb-1 cells were treated with 50 μmol/L pterostilbene for 12, 24 and 48 h. Pterostilbene induced cell apoptosis (P<0.01), the apoptosis rates of control group, 24 h treated group and 48 h treated group were (4.08±0.79)%, (13.44±2.12)% and (23.49±2.01)%. Pterostilbene induced autophagy of WERI-Rb-1 cells, increased LC3 expression, downregulated P62 expression and increased the number of autophagic vacuoles in WERI-Rb-1 cells (P<0.01). 3-MA and Beclin1 were able to rescue pterostilbene-induced cell death (P<0.01). After 3-MA was used to blunt autophagosome formation, the apoptosis rate markedly decreased in 3-MA+pterostilbene-treated cells compared with cells treated with pterostilbene alone [(12.97±2.09)%vs (8.35±1.11)%], and after siRNA was used to knockdown Beclin1, the apoptosis rate had the same change [(13.80±2.19)%vs (9.62±0.52)%].Conclusion:Pterostilbene can inhibit the proliferation of WERI-Rb-1 cells and induce cell apoptosis via autophagy activation.

Objective To investigate the relationship between the delivery gestational age, birth weight and fidgety movement. Methods 81 infants in fidgety movement phase were divided into preterm group and full-term group according to their delivery gestational age, into <2500 g group and ≥2500 g group according to their birth weight. The number of different kinds of General Movements (GMs) was compared.The correlation between birth weight, delivery of gestational age and GMs was analysed. Results The number of fidgety movement was more in the full-term group than in the preterm group (P=0.001), while the number of absence of fidgety movement was significantly less (P=0.003). The number of fidgety movement was more in the ≥2500 g group than in the <2500 g group (P=0.001), the number of absence of fidget movement was significantly less (P=0.003). The GMs was correlated with birth weight and delivery gestational age. Conclusion Premature and low birth weight are the risk factors for abnormal fidgety movement.

Phase Ⅰ Clinical trial of new drugs is key for clinical drug development. In this article,we describe our experience and reflection on phase Ⅰ clinical trials of new drugs at the First Affiliated Hospital of Nanjing Medical University.

Objective To study the ability of human embryonic stem cells to integrate into mouse liver and to repair chronic liver injury of the recipient.Methods On day 1,day-7 and day-15 after human embryonic stem cells were induced to differentiate into hepatocyte-like cells and were transplanted into mice with chronic liver failure,liver histopathology,liver function,liver tissue regeneration and mature hepatocytes of mice were tested respectively.Results Hepatic tissue pathology of mice significantly improved after transplantation and necrotic foci diminished,hemorrhage and congestion of hepatic cells relieved,and liver function improved.It was observed that human embryonic stem cells survived,proliferated,integrated with host liver,and differentiated into mature hepatocytes.Conclusions Human embryonic stem cells xenotransplanted into mice can participated liver tissue regeneration to some extent,and differentiated into functional liver cells.

Objective To investigate the influences of pain on early neonatal neurobehavioral development Methods 65 newborn infants admitted to the Neonatal Intensive Care Unit( NICU )of our hospital from October,2009 to March,2010 were randomly chosen as the objects of this study.In light of Neonatal Behavioral Neurological Assessment( NBNA) revised by Professor Bao Xiulan,examinations were carried out before and after pain stimulation,and a statistical analysis of the results of the examinations was conducted.Results The total scores of NBNA before and after the pain stimulation were (36.49±1.73) vs.(34.80±1.79) respectively,demonstrating a significant difference.Specifically,after the pain stimulation,the scores of behavioral ability and active muscle tension decreased,with a very significant difference.However,there was no significant difference in terms of the scores of passive muscle tone,primitive reflexes and common reactions.Conclusions Neonatal pain exerts influences on early neurobehavioral development,particularly on behavioral ability and active muscle tension.The training of neonatal health care professionals in the management of and the intervention in neonatal pain should be strengthened in order to decrease the adverse effects of pain on neonates.

Objective To investigate the influencing factors of pain and the changes of vital signs in newborn infants. Methods Forty two newborn infants were rated by the behavioral scale of acute pain in newborn infants. The scores of pain were compared among infants with different gender,gestational age,birth weight,birth age,type of puncture and whether by vaginal birth or not. At the same time,the respiration rate,heart rate,blood pressure and oxygen saturation (SO2)were dynami-cally recorded by the multi-function monitor in the process of puncture. Results The average score of pain was 7.6. There was no significant difference among newborn infants with different gender,gestational age,birth weight and type of puncture(P> 0.05),while significant differences among infants with different birth age and whether by vaginal birth or not (P=0.015 and 0.043 respectively). In the process of puncture,the SO2 was significantly decreased,while the respiration rate,heart rate,sys-tolic and diastolic blood pressure were significantly increased. Conclusions Pain is prevalent in newborn infants and accom-panied by obvious changes of vital signs. The means of childbirth and birth age have significant influence on the neonatal pain. It is suggested to pay close attention to the neonatal pain and take effective interventions.

BACKGROUND: Scar hypertrophy is always followed by the wound healing in burn and trauma. Endothelial cells play a key role in scar hypertrophy, so inhibitory growth of endothelial cells can relieve scar hypertrophy to a certain degree. OBJECTIVE: To construct a recombinant adenovirus vector expressing human endostatin (Ad/hEnd), and to investigate the cooperative effect of Ad/hEnd and keratinocyte on endothelial cell proliferation. DESIGN, TIME AND SETTING: Observational study, which was performed in the State Key Laboratory of Trauma, Burn and Combined Injury, Institute of Burn Research, Southwest Hospital of the Third Military Medical University of Chinese PLA between September 2006 and May 2007. MATERIALS: pAdTrack-CMV and pAdEasy-1 were obtained from Stratagene Company, USA; 293 cell and Ecoli.DH5α were stored in our laboratory. METHODS: The endostatin gene sequence was obtained by reverse transcription-polymerase chain reaction (RT-PCR) and polymerase chain reaction (PCR) based on mRNA of human fetal hepatic tissue and inserted into the adenovims shuttle plasmid pAdTrack-CMV to obtain recombinant plasmid pAdTrack-ES. After identification, positive recon was transformed into pAdeasy 1 recipient virus to screen positive clones. The adenovirus Ad/hEnd was generated from 293 cells and identified by PCR and fluorescence microscope. Then the keratinocytes were infected with Ad/hEnd, and co-cultured with endothelial cells by nest dish culture method. The content of endostatin was detected, and the non-transfection keratinocytes were used as the controls. MAIN OUTCOME MEASURES: Homologous recombination and identification of pAd/hEnd; generation and identification of Ad/hEnd; endostatin expression after 293 cell transfection; purification and titer measurement of Ad/hEnd; content of endostatin in culture solution; apoptotic percentage of endothelial cells; inhibitory ratio of endothelial cells. RESULTS: Ad/hEnd was constructed and the virus titer was generally up to 1.65×1012 PFU/L. Ad/hEnd-infected keratinocytes could effectively express and secrete endostatin of which the content reached 226 μg/L after 3 days of co-culture. The apoptotic percentage and inhibitory ratio of the endothelial cells co-cultured with Ad/hEnd-infected keratinocytes were significantly higher than those in control group (P<0.05). CONCLUSION: Ad/hEnd-infected keratinocytes co-cultured with endothelial cells can promote apoptosis and inhibit proliferation of endothelial cells through excretion of endostatin.

Objective To investigate the effects of the point-injection combined with neuromuscular facilitation rehabilitation techniques on post-stroke shoulder-hand syndrome. Methods A treatment group, 36 cases, was treated with the point-injection combined with neural facilitation of rehabilitation techniques. And a control group, 30 cases, was treated with massage therapy. Observed the clinical manifestations and used Fugl-Meyer to assess the joint activities, pain degree and motion function of upper limbs before and after therapy. Results Compared with the control group, the treatment group showed better improvement of joint activity scope and degree, and alleviation of pain (P<0.05). Conclusion Point injection and neuromuscular rehabilitation treatment has a better effect ain treating sequelare of brain stroke and it is worth applying.