Objective To construct the luciferase reporter gene vector of cell division cycle 2 (Cdc2) gene promoter and determine its transcriptional activity. Methods Primers were designed based on human Cdc2 promoter sequence from UCSC software. Then Cdc2 promoter from human genome DNA was replicated. After pGL3-Basic vector and Cdc2 promoter were digested with restriction enzymes SacⅠand XhoⅠseparately, Cdc2 promoter was inserted into pGL3-Basic vector. The recombinant plasmid named pGL3-Cdc2-promoter was transiently co-transfected into U2OS cells with control vector pRL-SV40, and then the activity of dual luciferase was detected. Results pGL3-Cdc2-promoter was constructed successfully. The restriction analysis and sequencing proved the entirely correct sequencing results. The luciferase activity was higher in pGL3-Cdc2-promoter/pRL-SV40 group than that of pGL3-Basic/pRL-SV40 group (1.591 5±0.199 8 vs 0.049 9±0.010 4). Conclusion pGL3-Cdc2-promoter can be transcribed and activated in U2OS cells. This study provided an important basis for screening and evaluation of anticancer drugs.

AIM: To investigate the changes of cardiac function, oxidative stress and apoptosis in myocardium of rat model of heart failure induced by adriamycin (ADR). METHODS: At the end of the study, we observed content of malondialdehyde (MDA), activity of superoxide dismutase (SOD) and apoptosis. Expression of P53 protein and p53 mRNA were measured with immunohistochemical method and RT-PCR, respectively. RESULTS: Our data showed that content of MDA increased, activity of SOD decreased and apoptosis in myocardium happened while cardiac function decreased after ADR treatment. p53 gene expression and P53 protein obviously increased in heart failure group. CONCLUSION: There were oxidative stress and apoptosis occurred significantly in a model of heart failure induced by ADR. p53 gene might play an important role in the apoptosis. Correlation analyses suggested that apoptosis in myocardium is related to oxidative stress in ADR-induced heart failure.

Objective To determine the levels of total antioxidative capacity(TAOC) and malondialdehyde(MDA) in liver exposed to hyperoxia,and to explore whether oxygen inhalation could cause liver injury in newborn rats.Methods Sixty-four newborn rats which were less than 12-hour-old were enrolled in this study.The rats were randomly divided into hyperoxia group(FiO2=0.85,n=32) and control group(air,n=32).Eight rats in each group were randomly sacrificed to obtain liver tissues at 1d,3d,7d and 14d.The TAOC of liver homogenates was detected by chemical colorimetry,and the MDA level of liver homogenates was measured by thiobarbituric acid test.Results In the hyperoxia group,TAOC in liver increased on the 1st day[(3.60±0.28)U/mg prot vs(3.39±0.19)U/mg prot,P<0.05];TAOC began to decreased on the 3rd day,and significantly lower than that of control group on the 14th day [(3.10±0.15)U/mg prot vs (3.56±0.14)U/mg prot,P<0.01].In the hyperoxia group,the MDA level increased on the 3rd day[(3.58±0.11)nmol/mg prot vs(2.82±0.14)nmol/mg prot,P<0.01],and reached a peak on the 7th day[(3.58 ±0.11)nmol/mg prot vs(2.82±0.14)nmol/mg prot,P<0.01],then decreased but still remained higher than control group on the 14th day [(2.92±0.18)nmol/mg prot vs(2.77 ±0.09)nmol/mg prot,P<0.01].Conclusion Too more MDA in liver and TAOC decrease may cause liver injury in newborn rats exposed to hyperoxia.With the oxygen inhalation time prolonging,the liver injury aggravation.

Objective To study CT features of thymoma,so that to improve the accuracy of CT diagnosis and differential diagnosis.Methods 31 cases of thymomas proved by surgery and pathology were examined with conventional CT scans.CT findings of thymoma were analyzed.Results The lesions in 27 cases(87.1%)were located in the anterior upper and middle mediastinum.There were benign lesion in 11,including mass-cardiovascular interface(MCI) with convex type(8 cases),flatness type(1 cases) and concave type(2 cases).20 cases were malignant lesion,including MCI with cast type(18 cases) and concave type(2 cases).Irregular invasion to adjacent organs was found in 11 cases,others included pericardiac effusion(n=6),pericardial and mediastinal invasion(n=2),pleural effusion(n=4),pneumonia(n=2),lung,bone,mediastinal lymphadens metastasis(n=2) and liver,pancreas metastasis(n=1).Conclusion CT scans is of significant value in diagnosis and differential diagnosis of thymoma.

Objcetive: To investigate the effect of resveratrol (RES)combined with cisplatin (DDP)on the cholangiocarcinoma cell line FRH-0201 in vitro. Methods: FRH-0201 cells were exposed to different concentrations of RES or (and)DDP. Cell morphological changes were observed by light and fluorescence microscopy. Inhibition of cell proliferation was measured by MTT and colony-forming assay. And the flow cytometry were performed to detect the cell apoptosis rate and cell cycle. Results: Resveratrol inhibited the proliferation of FRH-0201 cells in a time-and dose-dependent manner in the concentration of 5～320 ?mol/L and the IC50 values after the exposure of 24, 48, and 72 h were 55.35, 32.84, and 28.01 ?mol/L respectively. RES and DDP had synergetic effect in the inhibition of FRH-0201 cells. Conclusion: RES can inhibit the growth of FRH-0201 cells and has the synergetic effect with DDP.

Objective:To evaluate the effect of remote lifestyle intervention on the management of weight and related indicators in the overweight and obese population.Methods:A total of 400 individuals with overweight or obesity who participated in remote lifestyle intervention in the Health Management Department of Shenzhen People′s Hospital from May 2015 to December 2018 were included as the remote intervention group, and 400 individuals with overweight and obesity who matched their age and gender were selected as the control group. Dietician established individual WeChat groups with the remote intervention group, and the WeChat platform was used to conduct remote lifestyle intervention for them, including setting weight control goals, giving timely feedback to the food log based on photos, providing exercise guidance and psychological support. The control group received personalized diet and exercise prescriptions, but did not receive remote intervention. After adjusting the data by propensity score matching method, univariate and multivariate logistic regression models were used to analyze the management effect of weight and related indicators in the two groups after one year.Results:After one year of intervention, effective data were obtained from a total of 755 cases (371 cases in the remote intervention group and 384 cases in the control group), and effective data were retained from 446 cases (223 cases in the remote intervention group and 223 cases in the control group) after bias matching. The body mass index (BMI), systolic blood pressure, fasting blood glucose, triglyceride, total cholesterol, low-density lipoprotein, and serum uric acid of the remote intervention group [(24.85±2.52) kg/m 2, (110.21±10.53) mmHg, (4.96±0.65) mmol/L, (1.25±0.82) mmol/L, (4.87±1.11) mmol/L, (2.88±0.74) mmol/L, and (306.01±95.66) mmol/L respectively] were significantly lower than that of the control group [(27.76±2.28) kg/m 2, (121.14±14.07) mmHg, (5.10±0.87) mmol/L, (1.54±0.83) mmol/L, (5.28±0.96) mmol/L, (3.13±0.80) mmol/L, (355.16±92.68) mmol/L respectively] (all P<0.05). After intervention, intervention was consistently being influencing factors when BMI was reduced by 4%―12%, ( P<0.05). The probability of a 12% reduction in BMI in the remote intervention group was 112.486 times higher than that in the control group (95% CI: 16.852-890.266). At the same time, the initial BMI was an influential factor for the restoration of normal BMI. For every 1 kg/m 2 decrease in the initial BMI, the probability of restoration of normal BMI was 4.76 times higher than that before the decrease (95% CI: 3.222-5.057). Conclusions:Remote lifestyle intervention has a certain effect on the management of weight and related indicators in the overweight and obese populations. It has significant effect on weight loss of overweight and mildly obese people, but has limited effect on moderate and severe obese people.

Objective To establish a nerve stem cell model possessing the potentiality of differentiating intoγ-aminobutyric acid neuron-like cells(GABAergic-like cells),and provide eligible investigative vector for study on GABAergic neurons degenerative diseases. Methods Dibutyryl cyclic adenosine monophosphate (dbcAMP)was applied to induce human neuroblastoma SH-SY5Y cells,and the SH-SY5Y cells were divided into control group (0 mmol·L-1 dbcAMP),0.3 mmol·L-1 dbcAMP group,0.6 mmol·L-1 dbcAMP group,1.0 mmol·L-1 dbcAMP group and 2.0 mmol·L-1 dbcAMP group;the morphological changes of SH-SY5Y cells were observed.The Image-Pro Plus 5.0 software was used to measure the length of neurites of the neuron-like cells, and the percentage of the SH-SY5Y cells with neurites longer than 30 μm was calculated. The immunofluorescence cytochemistry technique was utilized to test GAD65 positive cells,and the positive rate of immune response was calculated.Results The results of light microscope observation showed that the cells in control group were polygonal,circular or shuttle type with smooth membrane and clear boundaries. With the increasing of the concentrations of dbcAMP and the prolongation of time,the morphology of SH-SY5Y cells’bodies became smaller with longer processes in dbcAMP groups.The cells interwined each other and showed mature neuron phenotype in 1.0 mmol·L-1 dbcAMP group.Compared with control group(31.4%±4.2%),the percentages of the cell with neurites longer than 30 μm in 0.3, 0.6, 1.0, and 2.0 dbcAMP groups (40.1%± 5.7%, 47.5%± 6.2%, 73.1%±3.2%,and 74.3%± 6.1%)72 h after induction were significantly increased(P<0.05 or P<0.01). Compared with control group (10.2%± 2.1%), the GAD65 positive expression rates in 0.3, 0.6, 1.0,and 2.0 mmol·L-1,dbcAMP groups(22.1%±2.4%,46.9%±3.2%,70.7%±3.4%,and 72.3%±3.7%)72 h after induction were significantly increased(P<0.05 or P<0.01).Conclusion The SH-SY5Y cells have the potentiality of differentiating into GABAergic-like cells, and 1.0 mmol · L-1 is the optimal concentration of dbcAMP.

[ ABSTRACT] AIM:To investigate the combined effect of octreotide and Dachaihu decoction on the treatment of severe acute pancreatitis (SAP).METHODS:Wistar rats (n=50) were randomly divided into sham group, SAP group, octreotide group, Dachaihu decoction group and combination group.The quantity of ascites was measured.The levels of amylase, alanine aminotransferase and creatinine in the serum were examined.The morphological changes of the pancreatic tissues were observed by HE staining.The activation of NF-κB and IκBαexpression were determined by Western blot.The mRNA expression with ICAM-1 and IL-1 was detected by qPCR.RESULTS: Combined treatment with octreotide and Dachaihu decoction effectively reduced the quantity of ascites and the levels of amylase, alanine aminotransferase and creat-inine in the serum in SAP rats.Moreover, combined treatment significantly inhibited SAP-induced activation of NF-κB and decrease in IκBαprotein expression, accompanied by a decrease in ICAM-1 and IL-1 mRNA expression.CONCLU-SION:Combination of octreotide with Dachaihu decoction effectively attenuates SAP by inhibiting NF-κB signaling pathway and ICAM-1 and IL-1 expression.

reression of M 2 receptor mRNA of rat myocardium intoxicated by soman under high altitude hypoxia. Methods The expressions of M 2 receptor mRNA under hypoxia, soman intoxication and soman intoxication under hypoxia were detected by semi quantitative RT PCR, respectively. Results The expression of M 2 receptor mRNA increased in the high altitude hypoxia group. Both simple soman intoxication and combined soman intoxication and hypoxia decreased the expression of M 2 receptor rapidly. But under hypoxia, the expression increased significantly at 12 h and 24 h. Conclusion M 2 receptor was sensitive to nerve agents. Compared to simple soman intoxication group, the expression of M 2 receptor increased in combined soman intoxication and hypoxia group. This may be one of the major factors leading to aggravation of the injury of heart function by nerve agents in high altitude area.

Parkinson's disease(PD) is a common neurodegenerative disorder with no effective protective treatment,characterized by a massive degeneration of dopaminergic neurons in the substantia nigra(SNpc) and the subsequent loss of their projecting nerve fibers in the striatum.The major neurochemical manifestation of this disorder is the loss of the neurotransmitter dopamine(DA) in the striatum as a result of the progressive degeneration of the dopaminergic neurons in the substantia nigra.There have been significant progresses in recent years reporting on the use of mesenchymal stem cells(MSCs)in gene therapy,with specific application towards PD.MSCs,a kind of multipotent adult progenitor cells,are considered as a useful vehicle for cell and gene therapy because of their multiple differentiation potentiality and self-transplantation.The present study was focused on treating rat model of PD using human tyrosine hydroxylase gene(hTH),human aromatic L-amino acid decarboxylase gene(hAADC) and human GT Pcyclohydrolase I gene(hGCH-I) engineered MSCs,in order to provide a better understanding about the application of these cells in the therapeutic benifit of PD.The gene of hTH,hAADC and hGCH-I were introduced via recombinant adeno-associated virus(rAAV) infection into the MSCs in vitro.The genetically modified MSCs expressing hTH,hAADC and hGCH-I were transplanted into the striatum of PD rat models.The behavior,the nigra-striatal level of DA and its metabolite were detected.The results of present study were shown as follows:hTH,hAADC,hGCH-I and LacZ gene were transfected into MSCs with adeno-associated virus vectors.The HEK293 packaging cells(ATCC) were transfected with the plasmids of pAAV-hTH,pAAV-hAADC,pAAV-hGCH-I,pAAV-LacZ,pAAV-RC,pHelper by using calcium phosphate precipitation.Titer was detected using HT1080 cells.Viral particles were collected and used to infect MSCs.The purified modified MSCs expressing the three kinds of genes were selected separately and were grafted in the striatum of the PD model rats in the lesion side.The MSCs genetically modified suvived well 12 weeks after transplantation.The improvements of the behavior were observed every week after transplantation.Compared with the control group,the rounds of asymmetric rotation after apomorphine administration decreased in the groups double or triple genes engineered MSCs grafted(p