Objective To isolate the rat insulin gene enhancer binding protein 1(Islet-1)gene and construct plEGFP-C1-Islet-1 recombinant retroviral expression vector.Methods The cDNA encoding the rat Islet-1 gene was isolated by RT-PCR method,the cDNA was first cloned into PGET-1 TA vector to facilitate the sequence and then subcloned into the retroviral vector plEGFP-C1.plEGFP-C1-Islet-1 was transfected into PA317 packaging cells with lipofectamine 2000.Transformants were selected in medium containing G418.Results A 1 050 bp DNA fragment was obtained by RT-PCR;plEGFP-C1-Islet-1 recombinant retroviral expression vector was identified by restrictive enzymes digestion,PA317 cells transfected with recombinant vector expressed enhancer green fluorescent protein(EGFP).Conclusion The gene encoding the rat Islet-1 is obtained and plEGFP-C1-Islet-1 expression vector is constructed successfully.

Objective To study the effects of traditional Chinese medicine (TCM) five-tone therapy on nursing care of patients with irritable bowel syndrome (IBS).Methods By convenience sampling,a study was conducted among 100 patients with IBS.The patients were randomly divided into the control group and the intervention group,each group consisted of 50 patients.Patients in two groups were both treated with conventional drug and routine nursing care,but patients in the intervention group were intervened by TCM five-tone therapy for 3 months.Clinical efficiency of patients in two groups were assessed by Symptoms Check List-90 (SCL-90),Hamilton Anxiety Scale (HAMA) and Hamilton Depression Scale (HAMD)before treatment and after 12-week treatment.Results The clinical efficiency in the control group and intervention group were 60％ (30/50) and 84％ (42/50) after intervention (P＜0.05).The scores of HAMA and HAMD after TCM five-tone therapy intervened were (5.24±1.15)points and (5.21±1.63) points,which were lower than the control group (8.32±1.42)points and (7.64±1.96) points (P＜0.05).The intervention group was scored lower than the control group in SCL-90 (P＜0.05).Conclusions TCM five-tone therapy may be more effective in decreasing the depression and anxiety of patients with IBS and alleviating their abdominal symptoms.

Objective To explore the effects of traditional Chinese medicine (TCM) five-tone therapy in combination with constitutional types of Chinese medicine based dietary intervention on patients with irritable bowel syndrome (IBS). Methods By convenience sampling, a study was conducted among 68 patients with IBS. The patients were divided into the control group and the intervention group by random digital table method, and each group consisted of 34 patients. Patients in two groups were both treated with conventional drug and routine nursing care, but patients in the intervention group were provided TCM five-tone therapy in combination with constitutional types of Chinese medicine based dietary intervention for 3 months. Patients in two groups were assessed by Chinese Medicine Constitution Questionnaire (CMCQ), Symptom Check List-90 (SCL-90), Self-rating Anxiety Scale (SAS) and Self-rating Depression Scale (SDS) before treatment and after 12-week treatment. Results After TCM five-tone therapy in combination with constitutional types of Chinese medicine based dietary intervention, the number of IBS patients with pathological constitution types decreased in two groups, the number of patients with gentleness constitution increased to 8 and 25 separately in the control group and intervention group (P<0.05). After intervention, the scores of SAS and SDS in the intervention group were (40.73±8.27)points and (41.16±8.75)points respectively, which were lower than(48.36±7.53)points and(50.03±9.23)points of the control group, and the difference was statistically significant (P< 0.05). The intervention group scored lower than the control group in SCL-90, and the scores had significant differences between two groups (P<0.05). Conclusions TCM five-tone therapy in combination with constitutional types of Chinese medicine based dietary intervention may be more effective in adjusting pathological constitution to gentleness constitution of patients with IBS, alleviating their depression and anxiety and improving their physiological symptoms.

BACKGROUND: It has been suggested that amniotic epithelial cells (AECs) express almost all of the markers of neural cell and secret a lot of neurotrophic factors and neurotransmitters. If AECs could substitute neural cells, its neurotrophic effect will bring promising prospect in treating neuron injuries and degenerative neural disease.OBJECTIVE: To detect specific proteins of neural cells in rat's cultured AECs.DESIGN: Repeated measurement design.SETTING: Second Clinical Medical College , Jilin University; Department of Histology & Embryology, School of Basic Medical Science, Jilin University.MATERIALS: This experiment was conducted at the Department of Histology & Embryology, School of Basic Medical Science, Jilin University from October 2004 to October 2005. The rat amniotic epithelial tissue was mechanically peeled from an embryonic 12 to 14days Wistar rats. Mouse anti Nestin was purchased from Chemicon Co.,and anti-ChAT rabbit anti-NSE and anti-NT-3 antibodies from Wuhan Boshide Company. Mouse anti-Musashi antibody was donated by Pro.Okano.METHODS: AECs were dissociated and purified from the amnion of pregnancy 12-14 day rats. AECs were treated with trypsin for 5 minutes,then cultured in DMEM/F12 medium at a humidified atmosphere of 0.05 volume fraction of CO2 in air at 37 ℃. Cells were inoculated at a concentration of 5×109 cells/L in culture flask. After 3 days, cells were inoculated onto poly-lysine-treated 35 mm culture Petri dish at a density of 1 × 108 cells/L for immunocytochemically staining. The cells were fixed with 40 g/L paraformaldehyde for 20 minutes. Immunocytochemical staining method was used to detect the expression of microtubule-associated protein-2 (MAP-2),neuron specific enolase(NSE), glial fibrillary acidic protein (GFAP) and choline acetyl transferase(ChAT).MAIN OUTCOME MEASURES: ① Morphological observation of rat'AECs at different culture time. ② Expression of specific protein of neural cells in rat' cultured AECs.RESULTS: ① After cultured for 24 hours, the AECs were flat and presented fibroblast-like morphology. 3 to 5 days later, cell bodies were well stacked; AECs had a big and round nucleus and were connected with each other by flourishing dendrites. ② Immunocytochemical staining results after culture for 4 days showed that AECs expressed Nestin, ChAT,NSE, Musashi, MAP-2, GFAP.CONCLUSION: AECs are homologous to neural cells in morphology, and it may be a new cell source to treat nervous system disease.

Objective To study the effects of LaCl3 on P16 and P21 expressions of hepatocellular carcinoma cells.Methods The experiment was divided into 4 groups.In experiment groups,CBRH-7919 cells were cultivated with 0.01,0.10 and 1.00 mmol?L-1 LaCl3 in DMEM containing 10% calf serum.In control group,CBRH-7919 was cultivated in DMEM containing 10% calf serum without LaCl3.The growth of CBRH-7919 cells was observed following treated with 0.01,0.10,1.00 mmol?L-1 LaCl3 for 1,3,5 d in vitro, respectively,the changes of cell cycle were detected by flow cytometry.At the same time,P16 and P21 were detected by immunocytochemical analysis.Results Compared with control group,the inhibitory rate of growth of CBRH-7919 cells was increased evidently when treated with 0.1 and 1.00 mmol?L-1 LaCl3 for 3 and 5 d(P

Objective:To investigate the clinical significance of dynamic analysis of plasma brain natriuretic peptide in patients with dilated cardiomyopathy with chronic heart failure.Methods:Ninety patients with dilated cardiomyopathy with chronic heart failure admitted into our hospital firom March 2012 to March 2016 were divided into group A (20 cases),group B (38 cases),and group C (32 cases) according to the NYHA grading.The plasma BNP levels and LVEF,LA,LVEDD,and LVESD in the three groups were detected and compared.The correlation of plasma BNP and cardiac function and ultrasonic cardiogram indexes were analyzed.And the capability of plasma BNP and LVEF in diagnosis of patients were analyzed and compared.Results:The plasma BNP level in group C was markedly higher than that of group A and group B (P＜0.05),and that in group B was much higher than that of group A (P＜0.05).And LA in group C was significantly higher than that of group A (P＜0.05),while differences in LVEF,LVEDD,and LVESD were not obvious (P＞0.05).The plasma BNP was positively correlated to NYHA grading,but had no significant correlation with the LVEF,LVEDD,LVESD,and LA (P＞0.05).Based on results of receiver operating characteristic curve analysis,plasma BNP =523.5 pg/mL was the threshold value for identification of patients with NYHA Ⅲ and Ⅳ (AUC=0.901,P＜0.001),while LVEF had not the capability (AUC=0.392,P=0.276).Conclusion:Detection of plasma BNP level had important clinical significance on diagnosis,screening and cardiac functional grading of patients with dilated cardiomyopathy with chronic heart failure.

Objective To investigate the probabilities of brain-derived stem cells from fetal rats differentiating into neurons and astrocytes by velvet antler polypeptide(VAP) in vitro. Methods Neural stem cells from E12-14d rats were cultured for 7 days until neural stem cells (NSCs) aggregations were formed into neurospheres. The neurospheres were cultured at different concentrations of VAP, and immunocytochemistry was used to detect the differentiation of neural stem cells. Results The differentiated cells in 50?g/L VAP group are more than that in control group; the number of NSE positive cells in 50?g/L,100?g/L and 200?g/L groups is more than that in control group.Conclusion Neural stem cells can be successfully induced into neurons by VAP in vitro, which could provide a basis for regeneration of nerve system.;

Objective:To explore the chain mediating effect of escape motivation and flow experience between frustration and online game addiction.Methods:Stratified cluster sampling method was used to randomly select 740 students from 5 universities in Beijing.Frustrate mental state scale (FMSS), escape subscale of online game motivation scale (OGMS), game flow scale (GFS), and the game addiction subscale of different types of internet addiction scale (DTIAS) were used for investigation. SPSS 25.0 software and PROCESS macro program model 6 were used to analyze the data and test the mediation effect.Results:Frustration (62.94±15.84) was positively correlated with escape motivation(6.89±2.34), flow experience(20.36±7.38), and online game addiction(16.05±6.62) ( r=0.30, 0.19, 0.39, all P<0.01). Escape motivation was positively correlated with flow experience and online game addiction ( r=0.51, 0.50, both P<0.01), while flow experience was positively correlated with online game addiction ( r=0.51, all P<0.01). The direct effect of frustration on game addiction was 0.23 (95% CI=0.17-0.30). Frustration indirectly affects game addiction through two paths. The single mediating effect of escape motivation was 0.07 (95% CI=0.05-0.11), and the chain mediating effect of escape motivation and flow experience was 0.05 (95% CI=0.04-0.08). Conclusion:Frustration not only directly affects online game addiction, but also indirectly affects online game addiction through escape motivation and flow experience.

Objective To estimate the clinical effect of fuzhenghuayu capsule in treatment of early hepatocirrhosis by liver instantaneous elastic imaging. Methods Eighty patients with early hepatocirrhosis were divided into experiment group and control group according to the treatment method with 40 cases each. All the patients in 2 groups were given the same antiviral treatment with entecavir dispersible tablets, and the patients in experiment group combined with fuzhenghuayu capsule. All treatment lasted for 6 months. The liver stiffness measurement (LSM) was measured before treatment and 1, 3 and 6 months after treatment by liver instantaneous elasticity imaging. The serum hyaluronic acid was measured, and the Child-Pugh score was evaluated at the same time. Results The LSM, hyaluronic acid and Child-Pugh scores 6 months after treatment in experiment group were significantly lower than those in control group:(19.3 ± 0.9) kPa vs. (29.6 ± 1.3) kPa, (215.6 ± 59.3)μg/L vs. (344.4 ± 39.6)μg/L and (2.1 ± 1.3) scores vs. (3.9 ± 0.9) scores, and there were statistical differences (P<0.05). No obvious adverse reactions related to the use of fuzhenghuayu capsule were found during the course of treatment. Conclusions Liver instantaneous elasticity imaging can be used to evaluate and monitor early hepatocirrhosis. Fuzhenghuayu capsule on patients with early hepatocirrhosis has a certain degree of curative effect.

Objective:To discuss the effects of Rhodosin,the extractant of medical plants na med Rhodiola Sachinonsis A.Bor,on aging rats and the model rats of Alzheier′s d isease (AD).Methods:Step down and water maze tests were used to determine the content of ace tylcholine (Ach) and the activity of acetylcholine transferase (ChAT) of hippoca mpus,lipid peroxide (LPO) and activity of superoxide dismute (SOD) of the cerebr al cortex,and cerebellar cortex and spinal cord were observed respectively.At th e same time,the general light microscope and eletron microscope were used to obs erve the changes in morphology.Results:The Rhodosin as an antioxidant could enhance the content of Ach and the activity of ChAT,reduce the formation of LPO,and increase activity of SOD.The Rh odosin could facilitate metabolism,enhance activity,and inhibit degeneration of aging rat cells.Conclusion:Rhodosin had obvious anti-aging effects on aing rats and also had p reventive,protective,anti-dementia effects on experimental dementia.