Objective:To evaluate the efficacy and safety of dl-3-butylphthalide ( NBP) injection and soft capsules in the treat-ment of acute middle cerebral artery infarction. Methods:Sixty-one patients with acute cerebral infarction in the left middle cerebral artery in 72 hours of onset of ischemic stroke with score of 5-25 according to the national institutes of health stroke scale ( NIHSS) were randomly divided into the observation group (n=31) and the control group (n=30). The control group was treated with the routine treatment, while the observation group was sequentially treated with NBP injection and soft capsules additionally. The treatment course was 90 days. Before the treatment, the NIHSS score was evaluated in both groups to compare the neurologic impairment degree. After the treatment, the daily living skills assessment was performed by Barthel index ( BI) and modified Rankin score ( mRS) , and the ad-verse reactions were recorded. Results:Before the treatment, the NIHSS score in the two groups had no statistical significance ( P>0. 05). After the treatment, the BI in the observation group and the control group was (88. 55 ± 16. 74) and (70. 67 ± 26. 18), and mRS was (1. 87 ± 1. 02) and (2. 53 ± 1. 40), respectively, suggesting the observation group had more favorable outcome than the con-trol group (P≤0. 05). The incidence of adverse reactions had no significant difference between the groups. Conclusion: dl-3-Bu-tylphthalide sequential therapy should be regarded as an effective and safe method for acute cerebral infarction, which can improve the daily living skills and 90-day outcome of patients.

Purpose To study the malignant transformation after treating rat oval cell line ( WB-F344 ) with chemical carcinogen N-methyl-N′-nitro-N-nitrosoguanidine ( MNNG) . Methods WB-F344 cells were cultured with MNNG for severe times. The biological characteristics of induced cells were detected through the following methods:to check proliferation activity by flow cytometry analysis, to examine malignant transformation degree of induced cells by soft agar assay and tumor formation in NOD/SCID mice, and to investi-gate the transcriptional and protein levels of hepatocellular carcinoma marker GGT, GST-P by real time-PCR. Results Oval cells in-duced by MNNG showed changes in biological characteristics and malignant molecular markers. Conclusion Hepatic oval cells model is successfully established, which can be confirmed by tumor formation in NOD/SCID mice.

Objective To assess the methylation patterns in CpG islands of SPARC genes and its relationship with clinicopathological parameters. Methods Bisulfite treatment of genomie DNA and sequencing analysis was used to study methylation patterns in the CpG islands of SPARC genes in fresh tissues from 6 cases of chronic pancreatitis, 6 normal pancreatic tissues, 17 pancreatic adenocarcinoma and the cancer adjacent tissues, as well as 6 normaI blood samples for normal control, and compared the results with clinicopathological parameters. Results WBC DNA showed no methylation of SPARC gene CpG islands. The methylation rates in CpG islands of SPARC genes in pancreatic adenocarcinoma, the cancer adjacent tissues, chronic pancreatitis and normal pancreatic groups (2, 3, 4, 5, 6, 7 CpG sites) were 61.6%, 47.1%, 37.5%, 24.7%, respectively. The methylation rates in CpG islands (1, 8, 9, 10, 11, 12 sites) were 52.0%, 28.7%, 16.7% and 0. The difference were statistically significant between the pancreatic adenocarcinoma and chronic pancreatitis as well as normal pancreas groups (P＜0.001), and the difference were not statistically significant between the pancreatic adenocarcinoma and the cancer adjacent tissues. CpG hypermethylation were not related to risk factors such as smoking, alcohol, history of CP, the tumor size, differentiation and TNM staging, lymph node metastasis. Conclusions CpG in SPARC gene extron 1 was hypermethylated in pancreatic cancer, and this may be an early event in the development of pancreatic cancer.

BACKGROUNDGreat efforts have been made to search for the angiogenic inhibitors in avascular tissues. Several proteins isolated from cartilage have been proved to have anti-angiogenic or anti-tumour effects. Because cartilage contains a great amount of hyaluronic acid (HA) oligosaccharides and abundant HA binding proteins (HABP), therefore, we speculated that HABP might be one of the factors regulating vascularization in cartilage or anti-angiogenesis in tumours. The purpose of this research was to evaluate the effects of hyaluronan binding protein on inhibiting tumour growth both in vivo and vitro.

METHODSA unique protein termed human brain hyaluronan (HA) binding protein (b-HABP) was cloned from human brain cDNA library. MDA-435 human breast cancer cell line was chosen as a transfectant. The in vitro underlying mechanisms were investigated by determining the possibilities of MDA-435/b-HABP colony formation on soft agar, the effects of the transfectant on the proliferation of endothelial cells and the expression levels of caspase 3 and FasL from MDA-435/b-HABP. The in vivo study included tumour growth on the chorioallantoic membrane (CAM) of chicken embryos and nude mice.

RESULTSColony formation assay revealed that the colonies formed by MDA-435/b-HABP were greatly reduced compared to mock transfectants. The conditioned media from MDA-435/b-HABP inhibited the growth of endothelial cells in culture. Caspase 3 and FasL expressions were induced by MDA-435/b-HABP. The size of tumours of MDA-435/b-HABP in both CAM and nude mice was much smaller than that of MDA-435 alone.

CONCLUSIONSHuman brain hyaluronan binding protein (b-HABP) may represent a new kind of naturally existing anti-tumour substance. This brain-derived glycoprotein may block tumour growth by inducing apoptosis of cancer cells or by decreasing angiogenesis in tumour tissue via inhibiting proliferation of endothelial cells.

Animals ; Brain Chemistry ; Cartilage ; physiology ; Caspase 3 ; Caspases ; biosynthesis ; Cell Line, Tumor ; Chick Embryo ; Cloning, Molecular ; Endothelial Cells ; cytology ; Fas Ligand Protein ; Humans ; Hyaluronan Receptors ; genetics ; physiology ; Hyaluronic Acid ; metabolism ; Membrane Glycoproteins ; biosynthesis ; Mice ; Neoplasms, Experimental ; therapy ; Neovascularization, Pathologic ; prevention & control ; Transfection

Purpose:In order to investigate the effects of repair of arteriovenous fistula on heart function soas to widen the range of the indication of allogenetic transplantation of kidney. Methods :Repair of arteriovenousfistula was performed in 8 patients who have received long term hemodialysis and then complicated with repeatedattack of heart failure 5 months before renal transplantation. Results:There are 7 patients had their cardiac cham-bers returned to normal, and 1 returned to normal upper limit. Renal transplantation was successfully performedin 6 patients, but death occurred in 1 case resulting from liver function failure. Superacute rejection and sponta-neous rupture of the kidney occurred in 1 patient. Conclusions:It is considered that the repair of arteriovenous fis-tula before renal transplantation is beneficial to improvement of heart function in these patients so as to widen therange of operative indications and create favourable condition for renal transplantation and decrease complica-tions. Color doppler sonography is of great value in monitoring pre-and postoperative heart function.

OBJECTIVETo establish a high efficient human coagulation factor VIII (FVIII) eukaryotic stable expression system using lentiviral vector, and determine its biosafety.

METHODSLentiviral transfer plasmid carrying human B-domain-deleted FVIII(BDDhFVIII)-IRES-GFP(BDDhFVIII/pXZ9)or IRES-GFP(pXZ9) was constructed. Lentivirus particles were produced by transiently co-transfected 3-plasmids into 293FT cells and further concentrated via ultracentrifugation. CHO cells were infected, 72h later, the FVIII antigen (FVIII:Ag) concentration in the medium was examined by ELISA, the activity was detected via one stage coagulation,and the transcription of FVIII in the infected CHO cells was determined by RT-PCR.Virus infection ability in the medium and the gag gene in CHO cells were determined to evaluate the model's biosafety.

RESULTSLentiviral transfer plasmid BDDhFVIII-IRES-GFP(BDDhFVIII/pXZ9)carrying human B-domain-deleted FVIII or IRES-GFP (pXZ9) was successfully constructed, and high titer lentiviruses has been prepared. The lentivirus could infect CHO cells efficiently, after an additional 72 h, the FVIII:Ag concentration had up to (1724.9±283.7) mU/ml, the FVIII:C level increased to (10.58±1.55)%, and transcripts of BDDhFVIII mRNA could be measured by RT-PCR. Neither the gag gene nor the virus in the supernatant was detected.

CONCLUSIONLentivirus-mediated human coagulation factor VIII could be expressed efficiently in CHO cells. The system couldn't produce offspring virus, proving a good biosafety.

Animals ; CHO Cells ; Cricetinae ; Cricetulus ; Factor VIII ; genetics ; Gene Expression ; Genetic Vectors ; Humans ; Lentivirus ; genetics ; Plasmids ; Transfection

To investigate the effects of 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor simvastatin (SV) on proliferation, apoptosis and the PI3K/AKT signaling pathway in human acute monocytic leukemia cell line SHI-1. SHI-1 cells were incubated with different concentrations of SV (5, 10, 15 µmol/L). Otherwise, SHI-1 cells without any treatment were used as control. Cells in different groups were collected at 24, 48 and 72 h after incubation for further detection. MTT method was used to assay the growth inhibition rate and flow cytometry was used to detect the early stage apoptosis ratio. The human PI3K-AKT Signaling Pathway RT(2) Profiler(TM) PCR Array was used to detect the expression of 84 genes involved in PI3K-AKT signaling. The results indicated that the SV inhibited the proliferation and inducted the apoptosis of SHI-1 cells in time- and dose-dependent manners significantly. The growth inhibition rates of SHI-1 cells treated with 15 µmol/L SV for 24, 48 and 72 h were 26.82, 47.09 and 63.92, respectively; and their early stage apoptosis ratios were 5.75, 13.25 and 15.59, respectively. Compared with the control group, expression levels of 39 genes were changed in the group of 15 µmol/L SV at 48 h, among them 26 genes were down-regulated and 13 genes were up-regulated. It is concluded that the SV can inhibit proliferation and induce apoptosis of SHI-1 cells, and the mechanism may be associated with the changes of gene expression level in PI3K-AKT signaling pathway regulated by SV.
Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Down-Regulation ; Gene Expression Regulation, Leukemic ; Humans ; Leukemia, Monocytic, Acute ; metabolism ; Phosphatidylinositol 3-Kinases ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; Signal Transduction ; drug effects ; Simvastatin ; pharmacology

OBJECTIVETo understand the effect of pinacidil on rat myocardial Ca(2+)regulation.

METHODSAfter baseline measurement and a period of equilibrium, myocytes were randomly allocated to one of 4 treatment groups: Control group (8 myocytes): incubation in Lactate Ringer's solution at 24 degrees C for 2 hours; K group (8 myocytes): incubation in Lactate Ringer's solution containing 16 mmol/L potassium at 24 degrees C for 2 hours; K+P group (8 myocytes): incubation in Lactate Ringer's solution containing potassium 16 mmol/L and pinacidil 50 micromol/L at 24 degrees C for 2 hours; K+P+G group (8 myocytes): incubation in Lactate Ringer's solution containing potassium 16 mmol/L, pinacidil 50 micromol/L and glibenclamide 10 micromol/L at 24 degrees C for 2 hours. After each incubation, myocytes were resuspended in cell culture media at the same temperature and intracellular [Ca(2+)](i) and SR Ca(2+) release were measured.

RESULTSThe amplitude percent of [Ca(2+)](i) transient evoked by electrical stimulation in the K group was significantly decreased to 67.05% - 80.11% compared to 90.27% - 95.57% in the K+P group during reperfusion after ischemia (P<0.01). The percent amplitude of the [Ca(2+)](i) transient evoked by the rapid application of 10 mmol caffeine in the K group myocyte was approximately 112.00%+/-16.93% compared with that of the [Ca(2+)](i) transient evoked by electrical stimulation. However, in the K+P group myocyte the peak amplitude of the caffeine induced Ca(2+) release was 173.15%+/-26.01% compared with electrical stimulation (P<0.01). The duration of transient evoked by caffeine in K+P group (3.20+/-0.71 ms was significantly shorter than that in K group (3.93+/-0.46) ms (P<0.05).

CONCLUSIONCardioplegic arrest with simultaneous activation of KATP channels preserves rat myocardial Ca2+ by inducing sarcoplasmic reticulum Ca(2+) release and by alteration of Na(+)-Ca(2+) exchanger to better maintain [Ca(2+)](i) homeostasis.

Animals ; Calcium ; metabolism ; Female ; Heart ; drug effects ; Male ; Myocardium ; metabolism ; Pinacidil ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Sodium ; metabolism

Objective:This study aimed to evaluate the effect of enhanced external counterpulsation(EECP)on left ventricular function in elderly patients with coronary slow flow phenomenon(CSFP)using two-dimensional speckle tracking echocardiography(2D-STE).Methods:This prospective case-control study included 30 patients aged ≥60 years with no stenotic lesions in the coronary arteries but with slow blood flow phenomenon in more than one major coronary artery who were treated at the Department of Geriatrics, Qilu Hospital, Shandong University, between December 2017 and December 2018, and were divided into a medication group with 16 participants and a medication plus EECP group with 14 participants, using the numerical lottery method.Patients in the group treated with EECP received 6-week 36-h EECP therapy in addition to lifestyle modification and drug treatment.Fourteen patients with normal coronary blood flow served as the control group.Conventional echocardiography and 2D-STE were used to evaluate changes in left ventricular function in the CSF patients before and after drug treatment and EECP.Results:Compared with the control group before treatment, patients in the drug treatment group and the drug treatment plus EECP group showed a decrease in mitral annular early diastolic velocity( P<0.01), an increase in the ratio of peak mitral early diastolic blood flow velocity to the mean peak mitral annular early diastolic velocity( P<0.05), and a decrease in left ventricular longitudinal strain during systole( P<0.01), the longitudinal systolic myocardial strain rate( P<0.01)and the early diastolic longitudinal peak strain rate( P<0.01).There was no statistically significant difference in values from conventional echocardiographic parameters before and after treatment in CSF patients of the medication group(all P>0.05).In the group receiving EECP, there were statistically significant differences in pre-and post-treatment values in ventricular septal early diastolic velocity[(6.22 ± 0.64)cm/s vs.(6.69 ± 0.44)cm/s], lateral wall early diastolic velocity[(8.01±0.68)cm/s vs.(8.41±0.29)cm/s], mitral valve to mitral annulus early diastolic peak velocity ratio[(10.51±1.38) vs.(9.74±0.37)], longitudinal left ventricular systolic strain[(-16.21±0.46)% vs.(-16.80±0.48)%], left ventricular systolic longitudinal strain rate[(-1.29±0.03)s -1vs.(-1.35±0.04)s -1], and early diastolic longitudinal strain rate[(1.35±0.03)s -1vs.(1.40±0.03)s -1](t-values were -3.70、-2.74、2.23、10.25、12.30、-19.15, all P<0.05). Conclusions:2D-STE can evaluate subclinical myocardial dysfunction early and quantitatively in elderly patients with CSF, and objectively reflect changes in left ventricular function before and after clinical intervention with EECP.

Objective:To develop evidence-based appropriate type of entity for the Centers for Disease Control and Prevention in order to improve the performance of disease control and prevention system in China.Methods:This study firstly proposed the core functions and essential public health programs for the Centers for Disease Control and Prevention. Using Structured Decision-Making, this study presented three alternative options for type of entity to make essential public health programs succeed, and reviewed the pros and cons of alternative options based on 15 evaluation indicators. Questionnaires were distributed to 47 stakeholders between April 10 and April 20 of 2020. Finally, an appropriate type of entity for the Centers for Disease Control and Prevention was identified by using both qualitative and quantitative methods.Results:A total of 47 stakeholders ranked alternative options. Aggregated scores of each alternative option was computed after weighting each indicators. The results shows that Option 1(professional technical civil service organization)has the highest score (58.22).Conclusion:Professional technical civil service organization is appropriate type of entity for Centers for Disease Control and Prevention.