OBJECTIVE To explore the dyna mic change of lncRNA expression during lung carcino-genesis induced by urethane.METHODS A total of 40 BALB/c mice received weekly ip injection of urethane 1 g·kg -1 for four continuous weeks,mice were euthanized at 12th week or 24th week after the first urethane treat ment,respectively.The RNA of lung tissues were isolated and used for microarray analysis.Based on the results of the microarray we selected lncRNA-AK017233 for additional qPCR analysis in individual mouse.RESULTS The incidence of lung cancer were 85% and 100% at 12th week and 24th week after the first ad ministration of urethane,respectively.The multiplicity and dia meter of lung tu mors in 24 weeks treated group were statistically significant fro m those in 12 weeks treated group (P<0.01 ),and pathological analysis showed that tu mors were classifiable as moderately differ-entiated adenocarcino ma.Total of 26 Down-regulated lncRNAs in which lncRNA-AK017233 stand for the most down-regulated lncRNA were identified by microcarray analysis.qPCR detected that the lncRNA-AK017233 was significantly altered by 0.33 ti mes in lung tu mors of urethane treated group at 12th week, co mpared to parallel lung tissues in urethane treated group at 12th week.AK017233 expression of ure-thane treated group was significantly reduced by 0.22 ti mes at 24th week,co mpared to parallel lung tis-sues in urethane treated group at 24th week.CONCLUSION LncRNA-AK017233 was consistently down-regulated during urethane induced lung carcinogenesis.

Objective To investigate the anti-fibrosis mechanism from effects of difference in iron load levels on activation and apoptosis of hepatic stellate cells(HSCs). Methods According to the difference in iron load levels in HSCs,HSC-T6 cells were divided into four groups:blank control,iron deposition model,50μmol/L desferrioxamine and 25 μmol/L desferrioxamine groups. Quantitative polymerase chain reaction(PCR)was applied for the detection of collagen type Ⅰ and transforming growth factor-β1(TGF-β1)mRNA expressions of HSC-T6 cells. Immunohistochemical assay was used for the detection of α-smooth muscle actin(α-SMA)expression. The method of terminal deoxynucleotidyl transferase(TdT)-mediated dUTP nick end labeling(TUNEL)was used for the examination of apoptosis of HSC-T6 cells. Under electron microscope,the ultrastructures of HSC-T6 cells were observed. Results Compared with the blank control group,despite the TGF-β1 mRNA expression in iron deposition model group was increased,no statistical significant difference was seen(1.594±0.168 vs. 1.477±0.126, P>0.05),whereas collagen type I mRNA expression was significantly enhanced(1.354±0.076 vs. 1.197±0.104, P<0.01). Both 50μmol/L and 25μmol/L desferrioxamine could down-regulate collagen type I and TGF-β1 mRNA expressions,and the action of 50μmol/L desferrioxamine was superior to that of 25μmol/L desferrioxamine(collagen typeⅠmRNA:0.391±0.076 vs. 0.688±0.060,TGF-β1 mRNA:0.421±0.068 vs. 0.714±0.090,both P<0.01). Iron deposition could induce HSCs to expressα-SMA in great amount,while apoptosis could be seen scarcely in iron deposited HSCs. By desferrioxamine therapy,α-SMA expression of HSCs was decreased significantly,but some of the cells underwent apoptosis. Conclusion Different iron load levels inside HSCs can induce activation or apoptosis of the cells,showing that iron plays an important role in regulating the process of HSCs activation and apoptosis and revealing that desferrioxamine possesses the potential action for treatment of liver fibrosis.

OBJECTIVE:To establish the system of curriculum based on working process and occupation ability for pharmacy major in higher vocational colleges. METHODS:Investigation was conducted among medical practitioners from pharmaceutical companies,hospitals,pharmaceutical factories,scientific research institutions and other related professionals. RESULTS:150 ques-tionnaires were sent out,and 141 valid questionnaires were collected with effective recovery rate of 94.00%. Results of investiga-tion showed that respondents most valued graduates with interpersonal and communication skills,followed by professional skills and practical ability. They were mainly clinical application of drugs,pharmacological effects and adverse reactions of drugs in the pharmacology theory teaching,the mechanism of action of drugs were weakened. The ability of prescription distribution,symptoms inquiring and drugs recommending should be strengthened in the pharmacology theory practice teaching. More than half of the re-spondents thought that confirmatory tests were necessary to keep,which helped to train students’practical ability and deepen the understanding of the theory. Meanwhile,it was important to strengthen the students’communication with the patients or their fami-lies and doctors to cultivate the ability of acquiring professional knowledge. CONCLUSIONS:The investigation provides basis for the making of curriculum standards of pharmacology,through which teaching contents are selected,teaching methods are de-signed,and it makes the pharmacology course full of post applicability and provides better decision-making basis to meet the posi-tion requirements.

Objective To observe the effect of low temperature compound propofol on the changes of apoptosis protein Caspase-3,autophagy-related proteins Beclin-1 and LC3-Ⅱ and thier changes of hippocampal neurons.Methods The rats were randomly divided into blank control group (Group A),propofol group at low temperature (Group B) and chloral hydrate group at low temperature (Group C),Group B and C were treated with low temperature for 30 min.Then,each group was subjected to cardiac perfusion and decapitated brain to prepare rat hippocampal neuronal tissue samples.The expression of Caspase-3 was detected by immunohistochemistry,Beclin-1 and LC3-Ⅱ protein was detected by immunoblotting.The ultrastructural changes of neurons were observed by transmission electron microscopy.Results The expression of Caspase-3,Beclin-1 and LC3-Ⅱ protein in each group were statistically significant differences(P＜0.05).The results of transmission electron microscopy showed that the neurons in group B and C were changed in different degrees,especially in group C neuronal apoptosis is obvious.Conclusion Autophagy and apoptosis in existence still exist in low temperature condition,while propofol can reduce this damage and have better protective effect on neurons.

Objective To explore the outcome of allogeneic hematopoietic stem cell transplantation (allo-HSCT) for relapsed/refractory acute myeloid leukemia (AML) patients in nonremission (NR) status and its related risk factors.Methods Thirty-nine relapsed/refractory AML patients in NR status who received allo-HSCT between Jan.2006 and Dec.2016 were retrospectively analyzed.Major end points of study included overall survival (OS),disease free survival (DFS),and relapse rate.Results All patients achieved hematopoietic reconstitution.Median time to neutrophil and platelet engraftment was 13 (11 20) and 16 (10-58) days,respectively.Acute graft-versus-host disease (aGVHD) occurred in 25 (64.1％) patients,and 11 (31.4％) patients developed chronic GVHD.During a median follow-up period of 9.1 (1.6-93.8) months,11 (28.9％) cases survived,10(26.3％) survived without leukemia,and 21 (53.8％) relapsed.The estimated 2 year OS and DFS were 30.0％ ± 8.0％ and 26.7％ ± 7.7％,and cumulative incidence of relapse and transplantation related mortality at 2 years was 56.63％ (95％ CI 37.84％-71.71％),19.7％ (95％ CI 8.3％-34.5％),respectively.The multivariate analysis revealed that the number of bone marrow blasts≥25％ or any level of blasts in peripheral blood was significantly associated with worse OS (HR =11.91,P=0.003),DFS (HR =10.75,P =0.002) and higher rate of relapse (70.83％ versus 20.22％,P =0.002).In addition,the development of grade Ⅱ-Ⅳ aGVHD also predicted an inferior OS (HR =3.18,P =0.039).Conclusion Salvage therapy with allo-HSCT can induce long-term survival in part refractory/relapsed AML patients.Decrease in the pre-transplant disease burden is the key to reduce relapse and improve survival.

Objective·To analyze the epidemiological characteristics of risk factors for cardiovascular diseases and malignant tumors based on the Shanghai community elderly cohort.Methods·The study subjects were selected from the Shanghai community elderly cohort established from February to August 2019,with a total of 17 948 people.The study subjects were divided into 4 groups according to self-reported presence or absence of tumors and/or cardiovascular diseases during the baseline survey:tumor-free and non-cardiovascular disease group,single cardiovascular disease group,single tumor group and tumor cardiovascular disease co-occurrence group.The differences among the four groups of subjects were collected and compared in terms of demographic characteristics and physiological indicators,daily living habits(smoking,drinking tea,drinking coffee,drinking carbonated drink,drinking alcohol,sedentary time,physical activity level and sleep quality),past medical history,psychological status(depression and anxiety)and dietary compliance.Results·Among the study subjects,60.1%of tumor patients were complicated with cardiovascular diseases.The differences among the four groups of subjects in age,gender,educational level,pre-retirement occupation,waist circumference,hip circumference and body mass index were statistically significant(all P<0.05).Compared with the tumor-free and non-cardiovascular disease group,the single cardiovascular disease group,single tumor group and tumor cardiovascular disease co-occurrence group all exhibited lower proportions of smoking and high physical activity levels(all P<0.05),and higher proportion of sedentary time exceeding 4 h/d and poor sleep quality(all P<0.05);the proportion of subjects with past medical histories including hyperlipidemia,peripheral vascular disease,endocrine system disease,respiratory system disease,urinary system disease and digestive system disease of the single cardiovascular disease group and the tumor cardiovascular disease co-occurrence group was higher(all P<0.05),and the proportion of subjects with depression and anxiety was also higher(all P<0.05).Furthermore,compared with the tumor-free and non-cardiovascular disease group,the single cardiovascular disease group had lower compliance rates of poultry,fish,fruit and liquid milk(all P<0.05).Among the four groups,only the compliance rate of vegetable intake exceeded 50%,while the compliance rates of poultry,fish,fruit,liquid milk and tubers were all below 20%.Conclusion·In the elderly population of Shanghai communities,over half of malignant tumor patients are concomitant with cardiovascular diseases.Unhealthy daily habits are prevalent among those with cardiovascular diseases,tumors and tumor-cardiovascular disease co-occurrence.The intake of many foods in the elderly of the community do not reach the levels recommended by Chinese Dietary Guidelines.

Alzheimer’s disease （AD）is a common latent neurodegenerative disease ，which is characterized by cognitive impairment，loss of learning and memory function ，abnormal behavior and dementia. At present ，there is no specific drug to effectively prevent or reverse AD. Gardenia jasminoides is the dried and mature fruit of G. jasminoides J. Ellis ，a gardenia plant in Rubiaceae. Its chemical components mainly include iridoids ，triterpenoids，organic acids and volatile oils ，among which iridoids are the main active components of G. jasminoides . This paper summarizes the researches on the mechanism of iridoids from G. jasminoides against AD at home and abroad in recent years ，in order to provide reference for the development of new drugs against AD.

Objective·To construct and verify the mouse model that can mimic the vitamin A deficiency(VAD)-like craniofacial skeletal deformity and do not cause embryonic death.Methods·Based on the Cre-LoxP system,the OsxCre;Rosa26dn/dn mice expressing osteoblast-specific dominant-negative retinoid acid receptor α(dnRARα)mutation were obtained by hybridization through OsxCre and Rosa26dnRARa/ddnRARa mice,to achieve the conditional inhibition of retinoic acid signaling to simulate VAD disease.Femur bone mesenchymal stem cells(BMSCs)and parietal bone cells of OsxCre;Rosa26dn/dn mice and their control littermates were isolated and underwent osteogenic induction,to assess the expression of retinoid acid receptor α(RARα)protein through Western blotting.Osteoblasts induced from parietal bone cells of OsxCre;Rosa26dn/dn mice and their control littermates were isolated and the effect of retinoic acid signaling inhibition was verified through dual luciferase gene reporter assay.Meanwhile,Ad-eGFP or Ad-Cre adenovirus-infected femur BMSCs and parietal bone cells of Rosa26dn/dnmice underwent osteogenic induction to assess the expression of dominant-negative mutant protein and the inhibition of the retinoic acid signaling pathway in vitro by Western blotting and dual luciferase gene reporter assay.Moreover,the skulls of 6-week-old OsxCre;Rosa26dn/dn mice were collected,and Micro-CT scanning and three-dimensional(3D)reconstruction were performed to verify the craniofacial skeletal deformities of the mouse model.Results·Western blotting results demonstrated that the level of RARα protein increased in the femur and parietal osteoblasts of OsxCre;Rosa26dn/dn mice compared to that of their control littermates,and also increased in the Ad-Cre-infected femur and parietal osteoblasts of Rosa26dn/dn mice compared to that in the Ad-eGFP-infected group(P＜0.05).Dualluciferase gene reporter assay results indicated that the activity of retinoid acid response element(RARE)was inhibited in the osteoblasts of OsxCre;Rosa26dn/dn mice compared to their control littermates,and was also inhibited in the Ad-Cre-infected group compared to the Ad-eGFP-infected group(P＜0.05).Micro-CT and 3D reconstruction suggested that the skull of 6-week-old OsxCre;Rosa26dn/dn mice exhibited VAD-like craniofacial skeletal deformities,including smaller size of the skull and osteogenesis imperfecta compared to their control littermates.Conclusion·An osteoblast-specific dnRARα expressing mouse model that can mimic VAD-like craniofacial skeletal deformity is successfully constructed,therefore providing a new model for exploring the pathogenesis and therapeutic targets of VAD-like craniofacial skeletal deformity in the future.

Objective·To explore the effect of all-trans retinoic acid(ATRA)of different concentrations on osteogenic differentiation of jaw bone mesenchymal stem cells(jBMSCs)in rats.Methods·jBMSCs from 4-week-old Sprague-Dawley(SD)rats were isolated and cultured with whole bone marrow adherence method.The surface antigens were identified by using flow cytometry.Alkaline phosphatase(ALP)staining/alizarin red staining,oil red O staining and alcian blue staining were used to prove the multilineage differentiation potential of jBMSCs after osteogenic,adipogenic and chondrogenic induction respectively.jBMSCs were induced in osteogenic medium with ATRA of concentration of 0.01,0.1,1,5,10,20 μmol/L in vitro,and dimethyl sulfoxide(DMSO)was used as control group.Cell viability of jBMSCs in different groups were determined by CCK8.ALP staining and alizarin red staining were used to investigate the osteogenic ability of jBMSCs in each group and screened the concentrations for subsequent experiments.Quantitative real-time polymerase chain reaction(qPCR)and immunofluorescence staining were used to analyze the expressions of osteogenesis-related genes and proteins in jBMSCs of different concentrations.Results·The flow cytometry analysis showed that more than 98%of P1 jBMSCs were positive for CD29+CD90+CD31-CD45-,which was congruent with the characteristics of bone mesenchymal stem cells.The results of ALP staining/alizarin red staining,oil red O staining and alcian blue staining indicated that the P1 jBMSCs had the multilineage differentiation potential of osteogenesis,adipogenesis and chondrogenesis.The results of ALP staining/alizarin red staining showed that the osteogenic activity and mineralization ability of jBMSCs in 0.01,0.1 and 1 μmol/L ATRA groups were increased compared with those in the control group,while the osteogenic activity and mineralization ability were decreased when the concentration of ATRA increased,especially higher than 5 μmol/L(all P<0.05).qPCR analysis showed that the mRNA expression levels of osteogenesis-related genes such as Alp,bone sialoprotein(Bsp),collagen type Ⅰ α1(Col1a1)and osteocalcin(Ocn)were higher in the 0.1 and 1 μmol/L ATRA groups compared to the control group.However,further increasing the concentration of ATRA led to a decrease in gene expression levels,and when the concentration exceeded 5 μmol/L,it began to be lower than the control group level(all P<0.05).The immunofluorescence staining showed that the expression of osteogenic related proteins SP7,ALP and OCN in the 0.1 and 1 μmol/L ATRA groups were increased compared to the control group,while further increasing the concentration of ATRA led to a decrease in protein expression.When the concentration was higher than 5 μmol/L,it began to be lower than the control group level(all P<0.05).Conclusion·Lower concentrations(0.1,1 μmol/L)of ATRA can promote the osteogenic differentiation of rat jBMSCs,and the promoting effect reaches its peak at 0.1 μmol/L,while the effect can be weakened by further increasing the concentration.Higher concentrations(5,10,20 μmol/L)of ATRA could inhibit the osteogenic differentiation of rat jBMSCs,showing an inhibitory effect.In this study,the dual-directional effect of retinoic acid on osteogenic differentiation of jBMSCs was demonstrated in vitro,and 0.1 μmol/L ATRA was identified as the optimal concentration for osteogenic differentiation of jBMSCs in rats,which provided a reference basis for the development of in vivo studies and clinical application of ATRA.