OBJECTIVE To develop a long-acting light-protective nanogel with both physical barrier and chemical clearance functions， and evaluate its performance. METHODS The photoprotective nanogel composed of mussel mucin and sodium hyaluronate was constructed based on a fullerenol-cerium oxide composite nano system， namely fullerenol-cerium oxide nanogel （FCN）， and was characterized. The antioxidant capacity of FCN was evaluated using in vitro free radical scavenging experiments； its UV shielding ability was assessed by using an SPF value detector； its biosafety was assessed according to the requirements of the Guidelines for Drug Safety Evaluation； skin adhesion was assessed using small animal 3D live imaging technology； its sun protection ability was assessed through skin sunscreen detection and histopathological observation. RESULTS The average particle sizes of cerium oxide and fullerenol nanoparticles in FCN were about 20 and 10 nm， respectively， and FCN exhibited good UV absorption and free radical scavenging abilities. SPF value of FCN was 58.95±0.82， and the ultraviolet A protection level value was 6.21±0.15. No pathogenic colonies such as Staphylococcus aureus， were detected in the nanogel， and the contents of lead， arsenic， mercury and cadmium all met the standards for pharmaceutical excipients； FCN group did not show any irritating reactions such as erythema， edema， or desquamation； blood biochemical indicators of the FCN group were within the normal reference range. The material clearance rate of mice in the artificial sweat flushing group was less than 30%， while the material clearance rate of mice in the dry cleaning group reached about 92%. The mice in the protective group did not show obvious erythema or ulcer formation throughout the experiment. Histopathology showed that the fibers were arranged in an orderly manner， and the number of collagen fibers was close to that of the control group. CONCLUSIONS The FCN formulation constructed in this study meets the relevant requirements of the Chinese Pharmacopoeia， has good safety and skin compatibility， and achieves dual synergistic protection of UV shielding and free radical scavenging.

Objective:To investigate the application effect of high-altitude field-based teaching in acute mountain sickness.Methods:The medical students of the classes 2018 and 2019 majoring in clinical medicine were selected as subjects, and they were divided into conventional teaching group and field-based teaching group, with 20 students in each group. The students in the conventional teaching group received classroom teaching alone, and those in the field-based teaching group received high-altitude field-based teaching after theoretical lectures. The two groups were compared in terms of the theoretical knowledge of acute mountain sickness, the quality score of internship, and rescue operation score of acute mountain sickness, and questionnaire feedback and post-class discussion were performed among trainees and teachers to evaluate the high-altitude field-based teaching model. SPSS 19.0 was used for statistical analysis.Results:Compared with the conventional teaching group, the field-based teaching group had significantly higher scores of the theoretical knowledge of acute mountain sickness (91.72±4.34 vs. 86.10±5.15, P<0.001), the quality score of internship (89.64±5.21 vs. 83.51±2.38, P<0.001), and the rescue operation of acute mountain sickness [94.05 (89.54, 94.87) vs. 87.01 (84.33, 90.82), P<0.001]. Conclusions:High-altitude field-based teaching can improve the teaching effect of acute mountain sickness and cultivate the interest and learning enthusiasm of students, and therefore, it holds promise for wide application.

Objective To explore the role and potential mechanisms of all-trans retinoic acid(ATRA)on activation and oxidative stress of hepatic stellate cell(HSC).Methods Platelet-derived growth factor(PDGF-bb,10 ng/ml)was applied to induce the activation of HSCs,which was then treated with ATRA at a dosage of 5 μmol/L for 48 h.The effects of ATRA on HSC activation were evaluated by detecting changes in cell growth viability and phenotypic marker expression.The effects of ATRA on HSC oxidative stress were evaluated by detecting changes in intracellular reactive oxygen species(ROS),reduced glutathione(GSH)and malondialdehyde(MDA),and the expression of antioxidant genes.The effects of ATRA on HSC autophagic activity were evaluated by detecting changes in autophagy marker expression and autophagic flow.Results Compared with the PDGF-bb group,the cell viability was significantly reduced in ATRA-treated HSCs(P＜0.01),as well as the expression of α-SMA and Collagen I.The intracellular levels of ROS and MDA were significantly reduced in ATRA-treated HSCs(P＜0.01),whereas the GSH level was significantly increased(P＜0.01).The expression levels of antioxidant genes(NRF2,HO-1,and ATF4),were significantly higher in ATRA-treated HSCs than those in the normal ones under PDGF-bb condition(P＜0.01).Meanwhile,the expression of autophagy markers Beclin 1 and LC3 Ⅱ/I,and signal of autophagy flow in ATRA-treated HSCs were found to be significantly reduced(P＜0.01).Conclusion ATRA significantly inhibited PDGF-bb-induced HSC activation and reduced the level of oxidative stress and autophagic activity of HSCs,which had potential applications in the prevention and treatment of liver fibrosis.

Objective To investigate the expression of(glutathione S-Transferase Omega-1,GSTO1)in cervical cancer tissue and its correlation with patient survival time,and to explore the impact of GSTO1 N-glycosylation on proliferation,migration,invasion,and epithelial-mesenchymal transition of cervical cancer.Methods By using immunohistochemistry,the expression levels of GSTO1 in tumor cells of 82 cervical cancer patients were detected,and the correlation between GSTO1 expression and clinical pathological features was analyzed.Kaplan-Meier meth-od was used to plot survival curves and evaluate the impact of GSTO1 expression in cervical cancer tissues on pa-tient survival time.Univariate and multivariate Cox regression analyses were performed to assess the independent prognostic factors influencing cervical cancer prognosis.The NetNGlyc 1.0 Server database predicted potential N-glycosylation modification sites of GSTO1(Asn55,Asnl35,Asn190).The cervical cancer cells(HeLa)were transfected with GSTO1 N-glycosylation site mutation vectors at positions 55,135,and 190,as well as GSTO1 wild-type vector and empty vector.Stable transfected cells were selected using puromycin.Western blot experi-ments were performed to assess the effectiveness of lentiviral interference.The effects of GSTO1 N-glycosylation site mutations on proliferation,migration,and invasion of HeLa cells were evaluated using EdU proliferation assay,wound healing assay,and Transwell assay.The effect of GSTO1 N-glycosylation site mutations on the epithelial-mesenchymal transition of HeLa cells was detected using the Western blot experiment.Results Immunohistochem-istry results revealed high expression of GSTO1 in cervical cancer tissues.The expression rate of GSTO1 was signifi-cantly higher in cervical cancer tissues with deep stromal invision≥1/2,lymphovascular space invasion,and lymph node metastasis(P＜0.05).Moreover,high expression of GSTO1 was associated with poorer overall surviv-al.After N-glycosylation site-specific mutation of GSTO1,the cell count of proliferation,migration,and invasion in HeLa cells significantly decreased(P＜0.05).The Western blot results showed that N-glycosylation site mutation of GSTO1 significantly inhibited the epithelial-mesenchymal transition of HeLa cells.Conclusion The expression of GSTO1 in cervical cancer tissues is associated with stromal infiltration depth,lymphovascular space invasion and lymph node metastasis,and it is also correlated with shorter patient survival time.Site-specific mutations in GSTO1 N-glycosylation significantly inhibit the proliferation,migration and epitheli al-mesenchymal transition of HeLa cells.

Immunotherapy combined with effective therapeutics such as chemotherapy and photodynamic therapy have been shown to be a successful strategy to activate anti-tumor immune responses for improved anticancer treatment. However, developing multifunctional biodegradable, biocompatible, low-toxic but highly efficient, and clinically available transformed nano-immunostimulants remains a challenge and is in great demand. Herein, we report and design of a novel carrier-free photo-chemotherapeutic nano-prodrug COS-BA/Ce6 NPs by combining three multifunctional components-a self-assembled natural small molecule betulinic acid (BA), a water-soluble chitosan oligosaccharide (COS), and a low toxic photosensitizer chlorin e6 (Ce6)-to augment the antitumor efficacy of the immune adjuvant anti-PD-L1-mediated cancer immunotherapy. We show that the designed nanodrugs harbored a smart and distinctive "dormancy" characteristic in chemotherapeutic effect with desired lower cytotoxicity, and multiple favorable therapeutic features including improved ¹O₂ generation induced by the reduced energy gap of Ce6, pH-responsiveness, good biodegradability, and biocompatibility, ensuring a highly efficient, synergistic photochemotherapy. Moreover, when combined with anti-PD-L1 therapy, both nano-coassembly based chemotherapy and chemotherapy/photodynamic therapy (PDT) could effectively activate antitumor immunity when treating primary or distant tumors, opening up potentially attractive possibilities for clinical immunotherapy.

Objective:To investigate the consistency between the results of serum specific antibodies to respiratory viruses and the nucleic acid tests in children with community acquired pneumonia, and to guide clinical application of these tests.Methods:Data of 109 cases of children with community acquired pneumonia diagnosed as viral infection by real-time fluorescence quantitative PCR from nasopharyngeal swab specimens were collected. Their acute and convalescent serum specimens were collected and used to detect common respiratory viruses (respiratory syncytial virus, adenovirus, influenza A virus, influenza B virus and parainfluenza virus) specific IgM antibodies by indirect immunofluorescence method . The consistency between the results of virus serum specific antibody test and respiratory virus nucleic acid test was analyzed.Results:Among the 109 cases of community-acquired pneumonia patients with positive respiratory virus nucleic acid test, 14 cases (12.8%) were positive for virus specific IgM in the acute stage. Combining the results of serum virus specific antibody detection in the acute and convalescent phases, 17 cases (15.6%) were positive for virus specific IgM antibody. Consistency analysis showed that the acute phase serum specific IgM antibody test results of respiratory syncytial virus, adenovirus, influenza A virus, influenza B virus and parainfluenza virus were compared with the nucleic acid test results of respiratory specimens, and Kappa values were -0.033, -0.003, 0.053, -0.024, -0.053, respectively. Compared with the nucleic acid test results of respiratory tract specimens and the total serum specific IgM test results of respiratory syncytial virus, adenovirus, influenza A virus, influenza B virus and parainfluenza virus in the acute and convalescence stages, Kappa values were 0.040, -0.053, 0.065, -0.030 and -0.018, respectively. Kappa values were all less than 0.4, indicating that the consistency of detection results of the two methods is poor.Conclusions:The results of serum virus specific IgM antibody detection in children with community acquired pneumonia are poorly consistent with the results of viral nucleic acid detection in respiratory specimens, suggesting that serum virus specific antibody detection with the immunofluorescence method may not be suitable for the diagnosis of viral pathogens in children’s community acquired pneumonia.

Objective:To explore the clinical application and long-term safety of hydroxychloroquine sulfate (HCQ) in the treatment of rheumatic diseases.Methods:A multi-center cross-sectional study was conducted between August 2017 and August 2018 in a random sample of eleven medical institutions of rheumatology and immunology in China. Patients who took HCQ for more than 3 months were enrolled into this study. The cumulative dose and long-term side effects of HCQ were recorded. The changes of laboratory indexes before and after treatment with HCQ were analyzed. Categorical variables were presented with counts and proportions, and evaluated by Chi-square test. Continuous parametric data were presented as Mean±standard deviation, and evaluated by Student's t test or Mann-Whitney U test. P-values less than 0.05 were considered statistically significant. Results:A total of 886 patients with rheumatic diseases were enrolled into this study, including 505 cases with systemic lupus erythematosus (57.0%), 210 cases with rheumatoid arthritis (23.7%), 80 cases with Sj?gren's syndrome (9.0%), 57 cases with undifferentiated connective tissue disease (6.4%), 12 cases of systemic vasculitis (1.4%), 10 cases of mixed connective tissue disease (1.1%), 7 cases of myositis (0.8%) and 5 cases with systemic sclerosis (0.6%). The most common long-term side effects of HCQ was skin or mucous lesions (12.4%) and vision problems (8.0%). Other adverse reactions included problems of digestive system (3.0%), nervous system (2.1%), musculoskeletal system (1.1%) and cardiovascular system (0.9%). 140 cases (15.8%) had stopped taking HCQ during the treatment. More than half of them decided to stop taking medicine by themselves. Fifty-four patients (6.1%) stopped using HCQ due to side effects while 24 of them took it again, and another 12 patients (1.4%) stopped the drug due to remission of illness. Patients were divided into three groups according to the cumulative dose of HCQ: less than 500 g, 500-1 000 g and more than 1 000 g respectively. There was significant difference in the incidence of long-term side effects among the three groups ( χ2=6.382, P=0.041). The last group (more than 1 000 g) suffered the highest incidence of long-term adverse reactions (37.1%). No severe adverse drug reactions were observed in this study. Conclusion:Hydroxychloroquine is widely used in the treatment of rheumatic diseases. The incidence of long-term side effects is 20.4%, is 6.1% lead to drug withdrawal, which are especially related to the cumulative doses. It should be adjusted properly according to the clinical application.

Objective: To investigate the diagnosis and treatment of synchronous double primary malignant tumours of the tongue and lung. Methods: A case of adenoid cystic carcinoma（ACC） and lung adenocarcinoma with double primary malignancy was retrospectively analyzed. Results: The tumor of patient′s tongue base gradually grew. MRI showed multiple enlarged lymph nodes on both sides of neck. CT of the chest showed obvious lesions in the anterior basal segment of the right lower lobe. The pathological biopsy of the tongue mass identified ACC, and pathological biopsy of the lung mass identified lung adenocarcinoma. The tongue and lung tumors were both surgically resected, and the tongue defect was repaired at the same time. No residue was found after surgery, and no recurrence was found during the follow-up period. The aesthetic and functional restoration of the lingual region was good. Conclusion There are few cases of adenoid cystic carcinoma and lung adenocarcinoma with double primary malignancies, and the related diagnosis and treatment are very difficult; the simultaneous removal of double primary malignant tumors may achieve good prognosis.

Objective: To investigate the impact of portal hypertention with hypersplenism of different severity and splenectomy on prognosis of hepatocellular carcinoma (HCC). Methods: We retrospectively analyzed the clinical data of 403 patients with HCC who met the Milan criteria and received radical treatment in Tianjin Third Central Hospital from January 2008 to January 2018. Cox propor-tional risk regression analysis was performed for parameters such as platelet levels (PLT), albumin-bilirubin (ALBI) grade, aspartate ami-notransferase-to-platelet ratio index (APRI), and post-sinusoidal resistance (PSR). HCC patients with severe hypersplenism were as-signed into two groups according to treatment method: radical treatment for HCC alone and radical treatment for HCC plus splenecto-my. Clinical data were compared, and the two groups were evaluated using the Kaplan-Meier survival analysis method. Results: Univar-iate and multivariate analyses showed that PLT was an independent risk factor for overall survival (OS) and disease-free survival (DFS) in patients with HCC. OS curves differed significantly with different PLT among patients with HCC (P=0.013). Furthermore, parameters of portal hypertension in cirrhosis, such as PSR, APRI, and ALBI grade, were risk factors for HCC prognosis. The degree of portal hyper-tension and hypersplenism, liver function, and tumor-node-metastasis stage did not differ between the two groups (P>0.05). Survival analysis showed significantly longer OS in the radical treatment plus splenectomy group (P=0.025). Following were the 1-, 3-, and 5-year survival rates: radical treatment alone group 100% , 98.2% , and 68.5% and radical treatment plus splenectomy group. 97.1% , 79.4%, and 56.8%, respectively. DFS did not differ between the two groups (P=0.326). Conclusions: Clinical parameters, such as PLT, PSR, APRI, and ALBI grade, are important prognostic factors in HCC patients with portal hypertension and hypersplenism. Radical treat-ment for HCC plus splenectomy can improve OS in HCC patients within the Milan criteria with severe hypersplenism.

The detection and molecular characterization of circulating tumor cells(CTCs) is one of the most important tool for liquid biopsy, which has the potential to enable non-invasive diagnostic tests for personalized medicine. Commercial platforms represented by CellSearch, the first FDA approved assay, have been considered to be valid for CTCs detection. However, special equipment and consumptive materials are required in the techniques listed above. Besides, most of them can not differentiate between apoptotic and viable cells, which indicates the portion of active and functional CTCs. Therefore, how to develop novel method for CTCs enrichment with metastatic potential has great significance in clinical routine. Telomerase-specific replication-selective oncolytic viruses expressing green fluorescent protein(GFP), including herpes simplex virus and adenovirus, allow the detection for human CTCs in the peripheral blood. After 24 h of transfection with recombinant virus, the tumor cells stably express GFP, and it could be used for CTCs counting by fluorescent microscopy or flow cytometry. Moreover, downstream analysis would be achieved by combination with PCR or DNA sequencing. Recombinant virus enables early detection of metastatic tumor cells, because the fluorescent signal is amplified only in viable, infected CTCs, by viral replication. This GFP-expressing virus-based method is remarkably sensitive, simple, and feasible, and it offers a new opportunity to detect and characterize CTCs in clinical routine.