Autophagy, a cellular housekeeping process, is indispensable to controlling the homeostasis of cytoplasm by removing unused proteins and damaged cell organelles. This process involves different types of human diseases, including cancers, neurodegenerative diseases and infectious diseases. Neurodegeneration is a critical pathological process of many eye diseases, such as glaucoma and age-related macular degeneration. The retina and all intraocular cells are constantly exposed to environmental stress and injuries, including oxidative stress and starvation, which lead to autophagy. Autophagy promotes cell survival through the recycling of metabolic precursors, or promotes cell death if autophagy is over-active. Additionally, autophagy and apoptosis have been shown to be harmonious or contrasting, depending on different experimental contexts. All of this contributes to the pathogenesis of many diseases. This paper reviews the mechanisms and regulation involved in autophagy, current understandings of neuronal autophagy in glaucoma and retina and strategies for therapeutic modulation.

BACKGROUND:It is a mature technology to culture MC3T3-E1 cels in the self-assembling peptide hydrogel, RADA16-NBD. Moreover, it is confirmed that a variety of metal ions, such as Fe, Cu, Zn, Mn, are involved in normal bone metabolism.
OBJECTIVE:To observe the effect of Cu2+and Fe3+ on the proliferation and differentiation of MC3T3-E1 cels cultured in the self-assembling peptide hydrogel, RADA16-NBD.
METHODS: Osteoblasts cultured with RADA16-NBD were divided into three groups and respectively cultured in culture medium containing Cu2+, Fe3+ or serum-free medium (control group), respectively. After 24, 48 and 72 hours, cel proliferation was detected by cel counting kit-8. After 1, 3, 5 days, alkaline phosphatase activity was detected. At 21 days, formation of calcified nodules was observed. Cel migration ability of cels was observed at 24 hours of Transwil chamber culture.
RESULTS AND CONCLUSION:Compared with the control group, the proliferative ability of cels cultured in the Cu2+, Fe3+ groups was significantly higher (P < 0.05,P < 0.01). At 72 hours of culture, there was no difference in the cel proliferation among the three groups. At 1, 3, 5 days of culture, the alkaline phosphatase activity in the Cu2+, Fe3+ groups was significantly higher than that in the control group (P < 0.05); while at 3 and 5 days of culture, the alkaline phosphatase activity in the Cu2+ group was significantly higher than that in the Fe3+ group (P < 0.05). In addition, the number of migrated cels was higher in the Cu2+ group than the Fe3+ group (P < 0.05). These findings indicate that both Cu2+ and Fe3+, especialy the former one, can promote MC3T3-E1 cel proliferation, differentiation and migration.

China ; Humans ; Lung Neoplasms ; diagnosis ; therapy ; Practice Guidelines as Topic ; standards

Children with pediatric tumors have better prognosis and longer survival than adults, suggesting that attention should be paid to the long-term complications induced by radiotherapy. In this article, the data from more than 40 clinical studies of pediatric tumor radiotherapy published in the recent decade were retrospectively analyzed. Long-term complications of nervous system, cardio-cerebrovascular system, respiratory system, endocrine system, urinary system, reproductive system, skeletal development, long-term secondary tumors were considered and the corresponding radiation dose-volume parameters were summarized, aiming to guide radiation oncology physicians and physicists to optimize radiotherapy plans for children with pediatric tumors.

Objective To measure serum levels of 25-hydroxyvitamin D (25HVD) in patients with chronic urticaria (CU),and to explore its role in the occurrence of CU.Methods Peripheral blood samples were obtained from 50 patients with CU and 40 healthy controls.The urticaria activity score (UAS) was used to assess the severity of CU and to group the patients with CU.Enzyme-linked immunosorbent assay (ELISA) was performed to measure the serum levels of 25HVD,interferon-γ (IFN-γ),interleukin-4 (IL-4) and immunoglobulin E (IgE).Statistical analysis was carried out by t test,rank sum test and linear correlation analysis.Results The serum level of 25HVD was significantly lower in the patients with CU than in the healthy controls (15.20 ± 7.72 vs.21.54 ± 8.31 μg/L,t =3.75,P ＜ 0.05).Moreover,there was a significant difference in the distribution of serum 25HVD levels between the patients and healthy controls (H =17.9,P ＜ 0.05).However,no significant difference was observed in the serum level of 25HVD between severe and mild CU groups (15.57 ± 7.38 vs.14.86 ± 6.28 μg/L,t =0.37,P ＞ 0.05).Compared with the control group,the patient group showed significantly decreased serum levels of IFN-γ (t =15.34,P ＜ 0.05),but increased serum levels of IL-4 and IgE (t =6.54,4.88,respectively,both P ＜ 0.05).Among the patients with CU,the serum level of 25HVD was positively correlated with that of IFN-γ(r =0.738,P ＜ 0.05),but negatively correlated with that of IL-4 (r =-0.689,P ＜ 0.05),and uncorrelated with that of IgE (r =-0.271,P ＞ 0.05).Conclusion The serum level of 25HVD evidently decreased in patients with CU,and it may participate in the occurrence of CU by mediating the Th 1/Th2 imbalance.

Objective To test the protective immunity in mice induced by recombinant Schistosoma japonicum Sj14FABP through several adjuvant formulations. Methods The recombinant Schistosoma japonicum Sj14FABP was prepared by expression in E. coli as a GST fusion protein (rSj14/GST) and used to vaccinate outbred Kunming mice by using complete Freund's adjuvant (FCA)/incomplete Freund's adjuvant (FIA), Bacillus Calmette-Guerin (BCG) and the immunostimulating complex (ISCOM) as adjuvant respectively. Results The purified recombinant protein rSj14/GST was immunogenic in mice, and 34.3% and 36.0% worm reduction rates were obtained in outbred Kunming mice immunized intradermally with BCG adjuvant and immunized subcutaneously with ISCOM adjuvant respectively, compared with non-vaccinated control group. However, intramuscularly vaccination with rSj14/GST in FCA/FIA was not protective, although the high level of IgG antibody was induced. Conclusion Both BCG and ISCOM are suitable adjuvants for rSj14/GST.

Objective To investigate the feature of the microorganisms colonization of the thoracic catheter-related infection and evaluate the clinical significance of prophylactic antibiotics administration in patients with pneumothorax treated with closed thoracic drainage. Method A total of 120 patients with pneumothorax treated with closed thoracic dramage in emergency department wore enrolled. The patients were randomized (random number) into group A (n =60) and group B (n =60). In group A, the patients received levofloxacin mesylate injection and in group B, patients received physiological saline injection instead after closed thoracic drainage. The tip of catheter was cut off to get a 2-cm long segment after catheter removal and this segment was dipped into a bottle filled with liquid culture medium for microorganism culture. Statistical analysis carried out by using χ2 test or Fisher exact test. Results Of all 120 patients, microorganisms were found in 49 segments of catheter and 57 strains of microorganisms were found. The four most common microorganisms were Coagulase-negative staphylococci (57.9%), Candida albicans (10. 5%),Staphylococcus aureus (7%) and Acinetobacter baumanii (7%). All of them were highly drug-resistant to β-1actam antibiotics. The difference in the positive rate of microorganism culture was distinct in pneumothorax patients with underlying diseases (50%) in comparison to the patients without underlying diseases (31%) (P ＜ 0.05). The positive rate of microorganism culture increased significantly as the duration of drainage was longer than 14 days (P ＜ 0.01). The positive rate of culture in group A was lower than that in group B if the duration of drainage was less than 7 days (8.3% vs 52.9%, P ＜ 0.01). The positive rate of culture after drainage for 7 days was 21.4% in group A and 68.8% in group B (P ＜0.05), and that after drainage for over 14 days was 70% in both groups (P ＞ 0.05). There were no significant differences in outcome and days of hospital stay between two groups (P ＞ 0. 05). Conclusions The common colonized microorganisms of thoracic catheter-related infection are conditional pathogens and highly resistant to antibiotics. Lengthening the duration of drainage and having underlying diseases increase the risk of infection. Although prophylactic antibiotics administration is beneficial to decrease the risk of thoracic catheter-related infection, it has no effects on shortening hospital stay and outcome of disease.

The gene fragment encoding the egg-shell precursor protein of Schistosoma japonicum was amplified with RT-PCR by using PCR primer designed according to the 423 bp cDNA fragment of the Philippine strain of S.japonicum, the corresponding mDNA fragment of Chinese strain as template and then the 5' and 3' ends of this gene cDNA were amplified with 5' RACE and 3' RACE by using a series of primers designed according to the result of sequencing. Result of sequence analysis showed that this fragment, named as Sj423, contained a complete open reading frame (ORF) of gene encoding the egg-shell precursor protein of S.japonicum.(Chinese strain). As demonstrated by sequencing analysis. No intron could be detected in this gene fragment. This gene was subsequently expressed in E.coli after cloning into the expression vector pET28c(+). The molecular mass of the expressed product of this gene was 20.9 kDa as revealed by SDS-PAGE analysis, and Western blot analysis showed that the recombinant protein expressed could react well with the rabbit antiserum against the worm antigen of S.japonicum;indicating the good antigenicity of this expressed product.

Objective To express the gene encoding mature protease of Schistosoma japonicum asparaginyl endopeptidase (Sj32) and evaluate the potential of the recombinant protein rSj32 in diagnosis of domestic animal schistosomiasis. Methods The DNA fragment encoding mature protease of Schistosoma japonicum asparaginyl endopeptidase was cloned with PCR from pET-28(a)/Sj32, and a recombinant plasmid was previously constructed in the laboratory, which contained the ORF of the gene encoding the pro-enzyme Sj32. The amplified DNA fragment was subcloned into pET-28a( + ) and the recombinant plasmid was transformed into E. coli BL21 (DE3) to express the mature protease of Sj32. Then the recombinant antigen (rSj32) was used in ELISA assay to diagnose schistosomiasis of mice, rabbits and water buffalo artificially infected. The detection effects of soluble Schistosoma japonicum egg antigen (SEA) , rSj32 and the recombinant 23 KDa membrane protein were compared. Results The recombinant antigen rSj32 with a molecular weight 41 KDa was successfully produced in E. coli BL21 ( DE3) and was purified with His Column with a yield of 25 mg/L E. coli culture. By using rSj32 as coating antigen in ELISA assay to detect the specific antibody in artificially infected mice, rabbits and buffalo, the sensitivities were 88.9% , 85.0% and 71.8% , respectively, the specificities were 100% , 96.7% and 96. 9% , respectively. There were no significant differences among the detection results of rSj32, SEA and rSj23. Conclusion rSj32 is a promising antigen for serological diagnosis of domestic animal schistosomiasis.

Objective　To compare the irradiation-protective and inter-synergestic effects of E838,WR-2721, Rubia cordifolia, cystamin e hydrochloride and ethinyl estradiol on radiation and combined radiation-burn injury. Methods　Above-mentioned drugs were given to the mice i ntraperitoneally, or intragastrcally, then, the mortality and the average surviv al d for 30 d were observed before and after the administration of the drug s. Results　①When drugs were before injury , the survival rate and the average survival d of the radiation and combined radiation-burn injured mice were increased obviously with the best effect in E838 and WR-2721. ②When drugs were given after injury, E838 and R. cordifolia also kept the effect. ③Combined appling WR-2721（pre) and E838（post）displayed a significant syner gistic reaction. Conclusion　E838 and WR-2721 are more e ffective than the others in the prevention of radiation.