Objective To investigate the impact of photodynamic therapy (PDT) with verteporfin on the expression of pigment epithelial derivative factor (PEDF) mRNA and vascular endothelial growth factor (VEGF) mRNA in adult retinal pigment epithelial (RPE) cells in vitro. Methods The changes of cellular viability before and after PDT were assessed by methyl thiazolyl tetrazolum (MTT) colorimetric assay. Semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) was conducted to detect the expression of PEDF and VEGF mRNA in RPE cells before and after PDT. Results PDT caused the death of RPE cells. The cellular mortality was positively correlated with the power of photocoagulation and the concentration of verteporfin. Conclusion PDT could down-regulate the expression of PEDF and VEGF mRNA in adult RPE cells in vitro, which may relate to the cure or relapse of subfoveal choroidal neovascular membrane after PDT.

cryopreservation periodontal tissue. METHODS:Periodontal tissue was scraped from healthy human teeth and divided them into three equal parts:fresh group, harvesting periodontal ligament stem cel s from fresh tissue;5%dimethyl sulfoxide group, 5%dimethyl sulfoxide added into the cryopreservation system;10%dimethyl sulfoxide group, 10%dimethyl sulfoxide added into the cryopreservation system. One month later, periodontal ligament stem cel s were extracted from the cryopreserved periodontal ligament from the latter two groups. RESULTS AND CONCLUSION:In the time that cel s swam out of tissue mass and cel harvest amount, the 5%dimethyl sulfoxide group was inferior to the fresh group but better than the 10%dimethyl sulfoxide group (P<0.05). No differences were found among the three groups in the fol owing aspects (P>0.05):colony formation rate of passage 1 periodontal ligament stem cel s, cel survival rate, proliferation ability of passage 3 periodontal ligament stem cel s, cel growth curve and surface marker expression of periodontal ligament stem cel s. The results suggest that the 5%dimethyl sulfoxide added cryopreserved system for periodontal ligament tissue cannot only shorten the time of periodontal ligament stem cel s amplification in vitro, ensure cel harvest and maintain basic cel ular biological characteristics, but also reduce the total amount of the dimethyl sulfoxide and the direct damage to the cel s caused by repeatedly frozen cel s, thereby providing a more secure guarantee for the future implementation of clinical transplantation therapy. So, the 5%dimethyl sulfoxide added cryopreserved system may be the new selection for donor tissue storage.

Mood disorders and substance use disorders often co-occur, but few evidence-based data have been conducted to guide the management of comorbidity bipolar disorder with alcohol use disorder.Therefore, it's lack of clinical researches and expert consensus.This review inducted the research progress of the comorbidities of bipolar disorder and alcohol use disorder in recent years, and finds that drug treatment or combined treatment with other treatment methods has certain curative effect suggesting that the comprehensive treatment of drug or combined non-drug treatment for comorbidities is the future direction.

Mood disorders and substance use disorders often co-occur,but few evidence-based data have been conducted to guide the management of comorbidity bipolar disorder with alcohol use disorder. Therefore,it's lack of clinical researches and expert consensus. This review inducted the research progress of the comorbidities of bipolar disorder and alcohol use disorder in recent years,and finds that drug treatment or combined treatment with other treatment methods has certain curative effect suggesting that the comprehen-sive treatment of drug or combined non-drug treatment for comorbidities is the future direction.

In recent years, cancer had become one of the main causes of death in children and adolescents, which had great adverse effects on their physiology and psychology, leading to serious physiological and psychological problems. Adventure based counseling had the effect of improving physical and mental health and quality of life, and had been widely used in the nursing of children with cancer. This paper reviewed the concept, intervention methods, applicable population and its application in the nursing of children and adolescents with cancer, in order to provide reference for the further research and application of adventure based counseling in children and adolescents with cancer in China.

Objective:To investigate the physiological role of F1Fo-ATP synthase α-subunit encoding gene (ATP1) in promoting Candida albicans ( C. albicans) to escape from macrophage killing through eliminating intracellular reactive oxygen species (ROS) by using a reverse genetics approach. Methods:An ATP1 deletion strain and a parental strain of C. albicans were cultured on the YPD media, and the number of formed colonies on the plates was counted to evaluate in vitro viability of C. albicans. To evaluate their in vivo viability, the ATP1 deletion strain and parental strain of C. albicans were inoculated into mice through the caudal vein, kidney tissues were taken out from the mice 1-7 days after the infection, and inoculated onto the YPD medium followed by numeration of colonies after 48 hours of culture. After co-culture of overnight-cultured C. albicans suspensions with macrophages, some of the C. albicans suspensions were inoculated onto the YPD solid medium followed by numeration of colonies and determination of survival rate, and some culture supernatants were inoculated into the 96-well plate for detection of the level of lactate dehydrogenase (LDH) released by macrophages by LDH release assay. A model mimicking oxidative stress in macrophages was established by using hydrogen peroxide. After treatment with hydrogen peroxide, the number of colonies was counted to compare the viability of the C. albicans strains. DCFH-DA staining was conducted to detect the intracellular ROS level in C. albicans after co-culture with macrophages or treatment with hydrogen peroxide, and real-time fluorescence-based quantitative PCR to measure mRNA expression of catalase 1 (CAT1) , superoxide dismutase 4 (SOD4) and SOD5 genes in C. albicans after treatment with hydrogen peroxide. Statistical analysis was carried out by using two-way analysis of variance or Student t test. Results:In vitro, the colony number in both the parental strain group and ATP1 deletion strain group gradually increased over time; after 24 hours, the colony number of the ATP1 deletion strain group was only 10% of that in the parental strain group ( F = 481.84, P < 0.001) . The number of colony formed by the parental strain-infected mouse kidney tissues gradually increased over time, but that by the ATP1 deletion strain-infected mouse kidney tissues gradually decreased, and there was a significant difference between the two groups ( F = 78.27, P = 0.001) . After in vitro co-culture of C. albicans with macrophages, the survival rate in the ATP1 deletion strain group (62.67% ± 3.51%) was significantly lower than that in the parental strain group (82.33% ± 2.52%, t = 7.88, P = 0.001) , and the percentage of LDH released by macrophages was also significantly lower in the ATP1 deletion strain group (27.80% ± 3.54%) than in the parental strain group (87.78% ± 0.17%, t = 33.89, P < 0.001) , which were consistent with the in vivo results. In the model mimicking oxidative stress, the viability of the ATP1 deletion strain group was significantly lower than that of the parental strain group ( F = 3 440.65, P < 0.001) . Both in the co-culture model with macrophages and in the model mimicking oxidative stress in macrophages, the intracellular ROS levels were significantly higher in the ATP1 deletion strain group than in the parental strain group (both P < 0.001) . Furthermore, the mRNA expression of CAT1, SOD4 and SOD5 genes was significantly lower in the ATP1 deletion strain group than in the parental strain group after treatment with hydrogen peroxide (all P < 0.001) . Conclusion:ATP1 deletion may reduce the capabilities of C. albicans to counteract oxidative stress and eliminate ROS, likely by down-regulating the expression of oxidative stress- and ROS clearance-related genes respectively, which may prevent C. albicans from escaping from the macrophage killing and lead it to be eliminated by the host ultimately.

By reviewing the necessity, current status of cognitive attitudes, influencing factors, and improvement strategies of inpatients’ living wills, this paper proposed to strengthen the publicity and promotion of living wills to advance the development of death with dignity, improve relevant influencing factors, and provide feasible solutions for living wills, and actively develop tools to assess the cognition and attitude of living wills, with a view to providing reference for further promoting the application of living wills in inpatients and improving related research.