Objective:To construct the human DcR3 expression vector and verify its expression in vitro. Methods: 915 bp human DcR3 gene CDS was amplified from porcine lung tissues,and was cloned into eukaryotic expression vector pEF1a-IRES-DsRed-Express2 which show red fluorescence. And then pEF1a-IRES-DsRed-Express2-DcR3 was transfected into LX-2 cells by FuGene HD. Expression of mRNA and protein lever of Human DcR3 were detected by RT-PCR and Western blot. Results:The levels of DcR3 gene transcription and translation in the hepatic stellate cells were significantly increased after transfection with pEF1a-IRES-DsRed-Ex-press2-DcR3 by RT-PCR and Western blot analysis. Conclusion: DcR3 expression vector was successfully constructed and highly expressed in LX-2 cells.

Refractory wounds are the common complication of diabetes,and the therapeutic scheme and methods are paid more and more attention nowadays. The influence of traditional Chinese medicines on diabetic wound healing, such as promoting fibroblasts proliferation, enhancing granulation tissue formation, increasing the secretion of growth factors and reducing the wound scar, was re-viewed in the paper to provide scientific basis and reference for traditional Chinese medicines used in diabetic refractory wounds.

Objective To investigate the effects of granulocyte macrophage colony-stimulating factor (GM-CSF) combined with interleukin-12 (IL-12) genes on apoptosis of hepatoma cells.Methods The hepatoma cell lines were cultured in vitro and were divided into four groups: GM-CSF transfection group,IL-12 transfection group,GM-CSF and IL-12 co-transfection group,negative control group (empty load group),respectively.The PIB-CMV3-GM-CSF and PIB-CMV3-IL-12 eukayotic expression vector was built,and 36 h after transfection,fluorescence microscope was used to detect the transfection effect;the expression level of IL-12,GM-CSF,p53,p38 and C-JUN mRNA were detected by RT-PCR,and Western blot was used to examine the expression level of IL-12,GM-CSF,p53,p38 and C-JUN protein.In addition,the flow cytometry was applied to detect cell apoptosis.Results Through fluorescence microscope,green fluorescence was observed in cells of GM-CSF transfection group,IL-12 transfection group,GM-CSF and IL-12 co-transfection group,indicating that the plasmid has successfully transferred into cells.In addition,the expression of p53mRNA in empty load group,GM-CSF transfection group,IL-12 transfection group,GM-CSF and IL-12 co-transfection group were 1.2±0.10,4.3±0.98,4.2±0.34,9.2±0.87,and the protein expression were 1.0±0.10,3.6±0.34,3.8±0.30,5.0±0.60.Compared with the empty load group,the expression level of p53 mRNA and protein were significantly increased in the three plasmid transfection groups (P<0.01).The expression of p53 mRNA and protein were significantly increased in co-transfection group than GM-CSF group and IL-12 group (P<0.01),while in the comparison with GM-CSF transfection group and IL-12 transfection group,the expression level of p53mRNA and protein in the co-transfection group could be improved to a higher degree(P<0.01).Meanwhile,p38 C-JUN mRNA expression levels in empty load group,GM-CSF transfection group,IL-12 transfection group,GM-CSF and IL-12 co-transfection group were as follows: 7.5± 0.9,3.5±0.45,3.7±0.25,1.0±0.11,while p38protein expression levels were 10.1±1.03,6.1± 0.67,7.1 ± 0.61,1.0 ± 0.12,respectively,C-JUN mRNA expression levels were 11.2 ± 1.20,4.1 ± 0.19,3.3 ± 0.30,1.0 ± 0.01,separately,C-JUN protein expression levels were 2.25 ± 0.2,1.8 ± 0.13,1.4 ± 0.12,1.0 ± 0.09.P38, C-JUN mRNA and protein levels were significantly reduced in the three plasmid transfection groups compared with the empty load group (P<0.01).The expression of p38,C-JUN mRNA and protein were reduced to a lower degree in co-transfection group than in GM-CSF transfection group and IL-12 transfection group (P<0.01).Flow cytometer showed that the hepatoma cell apoptosis rate of the empty load group,GM-CSF transfection group,IL-12 transfection group,co-transfection group were (3.43±0.9)%,(5.87±1.02)%,(7.32±1.1)%,(17.47±2.11)%,the rates of the three plasmid transfection groups were significantly higher than that of the empty load group (P<0.01).And the apoptosis rate was significantly increased in the co-transfected group compared with other plasmid groups (P<0.01). Conclusion The combination of GM-CSF and IL-12 could significantly accelerate the apoptosis of hepatoma cells by up-regulating the expression of p53,and down-regulating the expression of p38 and C-JUN.

Objective:To review and analyze the prescriptions for Chinese herbal pieces in an integrative traditional Chinese and western medicine hospital to explore the trend and problems in clinical application for Chinese herbal medicines and provide reference for dispensing in pharmacy of traditional medicine and rational drug use. Methods: Totally 1200 prescriptions for Chinese herbal pieces in our hospital were surveyed from January to December in 2014. The irrational use was analyzed, and reasonable suggestions were provided for the clinical application. Results:The problems in the prescriptions for Chinese herbal pieces included uncompleted diagnosis,incompatible diagnosis and medication,nonstandard Chinese name,unclear footnote,irrational usage or dosage and incompati-bility. The prescriptions including above 15 varieties of Chinese herbal pieces in one accounted for 65. 70%, and those with average dose above 250 g accounted for 40. 25%. Conclusion:Regular inspection and quality supervision on the prescription for Chinese herb-al pieces should be strengthened to improve the quality of prescriptions and guarantee the safety and economy of the medication.

ObjectiveTo investigate the effect of decoy receptor 3 (DcR3) gene on hepatocyte apoptosis, as well as the possible mechanism of action of DcR3 in this process. MethodsThe human liver cell lines were cultured in vitro, and pEF1α-DcR3 transfection group, pEF1α-IRES transfection group, and negative control group were established. The pEF1α-IRES-DsRed-Express2-DcR3 eukaryotic expression vector was constructed and transfected into human liver cell lines for 36 hours. qRT-PCR was used to measure the mRNA expression of DcR3, Fas ligand (FasL), α-smooth muscle actin (α-SMA), and transforming growth factor-β1 (TGF-β1), Western blot was used to measure the change in the protein expression of DcR3, and flow cytometry was used to measure apoptosis. An analysis of variance was used for comparison of continuous data between groups, and the least significant difference t-test was used for comparison between any two groups. ResultsAfter human liver cell lines were transfected with pEF1α-DcR3 for 36 hours, the pEF1α-DcR3 transfection group showed significant increases in the mRNA and protein expression of DcR3 compared with the pEF1α-IRES transfection group and negative control group (F=33 1695 and 14154, all P＜0.01). Compared with the other two groups, the pEF1α-DcR3 transfection group showed significant reductions in the mRNA expression of FasL, α-SMA, and TGF-β1 (F=269 4518, 20 7904, and 80678, all P＜0.01), which suggested that DcR3 inhibited the expression of FasL, α-SMA, and TGF-β1. Compared with the pEF1α-IRES transfection group and negative control group, the pEF1α-DcR3 transfection group showed a significant reduction in apoptosis rate (F=55863, all P＜0.01). ConclusionDcR3 can inhibit hepatocyte apoptosis and downregulate the mRNA expression of FasL, α-SMA, and TGF-β1.

The incidence of nonalcoholic fatty liver disease (NAFLD) has been increasing in China year by year, and NAFLD has become the second most common liver disease after viral hepatitis. Its development is closely associated with insulin resistance and oxidative stress and NAFLD is one of the important components of metabolic syndrome. Ghrelin is a newly discovered endogenous ligand for growth hormone secretagogue and has broad biological functions. Ghrelin not only affects the metabolic process of glucose and lipids, but also plays an important role in the development and progression of NAFLD. In this review, the role of endogenous Ghrelin expression in the development and progression of NAFLD is briefly summarized and the Ghrelin regulation in the development and progression of NAFLD is pointed out, which will provide new approaches to the clinical treatment of NAFLD.

Objective To explore a new efficient extraction method of dorsal root ganglions(DRGs)of rats by exploring and extracting DRGs reversely along the nerve traveling pathway. Methods The DRGs were extracted by the traditional method of opening the intervertebral foramina and the new method, extracting DRGs reversely along the nerve traveling pathway,respectively. The time consuming and the number of intact DRGs obtained with these two method were compared. Results The number of intact DRGs(L3-5segments,both sides)extracted from each rat with the traditional method was(3.08 ± 1.31),and the average time consuming of each DRG was(5.58 ± 1.21)min. As for the new method ,the number of intact DRGs extracted from each rat was(4.29 ± 1.08), and the average time consuming was(1.69 ± 0.91)min,significantly better than that of the traditional method(P < 0.05 for both). Conclusions The new method of exploring and extracting DRGs reversely along the nerve traveling pathway is more efficient for obtaining intact DRGs of rats,providing more useful tissue materials for subsequent culture and morphological studies of DRG cells.

OBJECTIVE: To improve the quality and efficiency of medication consultation and to promote rational drug use in clinic. METHODS: Totally of 520 cases of medication consultation records from the Affiliated Yueyang Hospital of Integrated Traditional Chinese and Western Medicine, Shanghai University of TCM during Oct. 2015-Oct. 2016 were selected according to random number table, and then summarized and analyzed statistically in respects of consultation content, age and type of consultants, department, consulted medicine types, the number of combination of TCM and Western medicine, satisfactory degree of consultants, etc. RESULTS: The main consultation contents included usage and dosage (41. 92％), combination of TCM and Western medicine (12. 88％), etc. Consultants mainly aged 50-70 years old (48. 27％), and were mainly the patients and their family members (95. 96％). Consultants were primarily diagnosed in cardiology department and endocrinology department. Top 3 consulted medicine types were cardiovascular system medicines, nervous system medicines, digestive system medicines, and types of Chinese patent medicines were mainly coordinating medicines and activating blood and removing stasis medicines. Types of TCM combined with Western medicine were less than or equal to 3 kinds in 420 cases (80. 77％). Most patients were very satisfied with outpatient medication consultation (93. 65％). CONCLUSIONS: Rational drug use consultation improves medication compliance of patients, meets the needs of patients on medication safety knowledge, and provides a strong guarantee for safe and rational drug use in clinic.

Objective: To explore mental health status of vocational college students and its association with physical exercise and exercise motivation, so as to provide data support for the promotion of physical exercise, exercise motivation theory and the protection of mental health among vocational college students. Methods: From June 2 to July 2, 2023, a convenience sampling method was used to select 1 763 college students from three vocational colleges in Sichuan, Chongqing, and Guizhou for the survey. Physical Activity Rating Scale (PARS-3), Motives for Physical Activities Measure-Revised (MPAM-R), Patient Health Questionnaire-9 (PHQ-9) and Flourishing Scale (FS) were used to assess physical exercise, exercise motivation, depressive symptoms and flourishing levels of vocational college students. Multiple linear regression analysis was conducted to examine the relationship between physical exercise, exercise motivation, depression as the negative indicator of mental health and flourishing as the positive indicator of mental health. Results: There were negative correlations between physical exercise, exercise motivation and their five factors with depression scores among vocational college students ( r=-0.162--0.133, P <0.01). Physical exercise, health motivation and pleasure motivation were positively correlated with flourishing and its eight factors among vocational college students ( r= 0.054 -0.099, P <0.05). Depression scores and flourishing scores varied significantly across students with different levels of physical exercise and exercise motivation ( F=11.18-15.69, 2.80-16.05, P <0.05). Multiple linear regression analysis indicated that physical exercise, health motivation, and total exercise motivation scores were negatively correlated with depressive symptoms ( β = -0.02 , -0.11, -0.35), and physical exercise, health motivation, pleasure motivation, and total exercise motivation scores were positively correlated with flourishing ( β =0.31, 0.32, 0.50, 0.30) ( P <0.05). Conclusions Physical exercise and exercise motivation are significantly associated with mental health of vocational college students. Enhancing physical exercise and increasing exercise motivation in aspects of health and pleasure can promote mental health of vocational college students.

In the field of bilingualism research, a key scientific question is how bilinguals process two language systems, particularly the effective switch from one language to another, namely bilingual code switching. With the rapid development of neuroimaging techniques, important progresses have been made in bilingual processing studies, especially in code switching. However, consensus has not been achieved regarding the mechanisms of bilingual code switching. Bilingual switching studies using neuropsychological and neuroimaging techniques have gained insights into the temporal and spatial features of the language switching process and the neurological mechanism, which provide direct evidence for the generation mechanism of bilingual code switching.