Background Age-related macular degeneration (AMD) is the leading cause of blindness in people over 50 years old,of which 90％ cases are caused by choroidal neovascularization (CNV).Current treatments on AMD have gained great achievements,but there are still some drawbacks.So we need to search for new targets to cure CNV.Objective This study was to construct two combined lentiviral vectors for rat Slit2 gene RNA interference (RNAi) and verify its interfering effects on Slit2 gene in rat retinal pigment epithelial (RPE) cells.Methods Two specific siRNA sequences targeting towards rat Slit2 gene were designed and were annealed to DNA sequences.The DNA sequences and GV248-enhanced green flourescent protein (EGFP) vectors were combined together as recombinant vectors and then were identified.The GV248-EGFP vector,helper 1.0 and helper 2.0 were transfected together into 293T cells and the two combined lentiviral vectors for rat Slit2 RNAi were gained from the cell supernatant after 72 hours of transfection.The titers of the combined lentiviral vectors were measured.The cells were divided into blank control group,Lv-EGFP vector group,Lv-rSlit2-siRNA1 group and Lv-rSlit2-siRNA2 group.The interference efficacy of the combined lentiviral vectors targeting to rat Slit2 gene were identified by real-time fluorescence quantitative PCR and Western blot.The sequence with higher interference efficacy was transfected to rat RPE cells again.The transfected and nontransfected rat RPE cells were treated with 0,100,200 and 400 μmol/L CoCl2 for the preparation of hypoxia models.The expression of vascular endothelial growth factor-A (VEGFA) mRNA in rat RPE cells was finally measured by real-time fluorescence quantitative PCR and the concentration of VEGFA protein in cell supernatant was assayed by ELISA.Results The recombined lentiviral vectors for rat Slit2 gene R NAi were successfully constructed.The titers of the two reconbinant sequences were 5× 108 TU/ml and 3×l08 TU/ml,with the transfected rate ≥70％.The relative expression levels of Slit2 mRNA in the Lv-rSlit2-siRNA1 group and LvrSlit2-siRNA2 group were 0.67±0.09 and 0.23±0.11,respectively,which were lower than 1.03±0.31 and 0.92± 0.07 in the blank control group and Lv-EGFP vector group (all at P＜0.01).The expression levels of Slit2 protein in the Lv-rSlit2-siRNA1 group and Lv-rSlit2-siRNA2 group were 0.62±0.07 and 0.49±0.02,respectively,which were lower than 1.00±0.10 in blank control group and 0.95±0.11 in Lv-EGFP vector group (all at P＜0.01).Significant differences were found in the expression of VEGFA mRNA and protein in RPE cells among different concentrations of CoC12 groups (mRNA:F tration =127.998,P＜0.01;Fgroup =69.663,P＜0.01.Protein:F ion =17.059,P＜ 0.01;Fgroup =91.791,P＜0.01),and the expression of VEGFA mRNA and the concentration of VEGFA protein were evidently lower in the Lv-rSlit2-siRNA2 group than those in the blank control group after being treated by 100,200 and 400 μmol/L CoCl2 (all at P＜0.01).Conclusions Recombined lentiviral vector for rat Slit2 gene RNAi is successfully constructed,which can effectively knockdown rat Slit2 gene and inhibit the expression of VEGFA in rat RPE cells.

Objective To investigate the treatment method of selective neck dissection(SND). Methods 68 cases patients with differentiated thyroid carcinoma were cured by selective neck dissection. Results The rate of lymph node diversion in 68 cases patients was 60.3%, among the diversion rate of lymph node in VI area was 51%. There were not patients who happend permanent damagement of laryngeal nerve and hypoparathyroidism. Conclusions It is available that selective neck dissection cures differentiated thyroid carcinoma.

0.05).The average length of hospitalization and average cost of interventional treatment group were less than those of operation drainage group(P

The existent problems, principles that should be observed and methods for English translation of TCM gynecological terms were discussed, and a scheme of translation for 150 TCM gynecological terms was explored.
Female ; Gynecology ; Humans ; Medicine, Chinese Traditional ; Terminology as Topic ; Translations

ObjectiveMyelodysplastic syndromes （MDS） is a group of clonal hematopoietic stem cell disorders，and this study aims to investigate the expression of hypoxia-inducible factor-1α（HIF-1α） in the bone marrow cells of patients with MDS and its correlation with the clinical features of MDS，the therapeutic efficacy of arsenic-containing Chineseherbal compound，and the survival prognosis. MethodAccording to the inclusion and exclusion criteria，27 MDS patients treated with arsenic-containing Chinese herbal compound in the Department of Hematology，Xiyuan Hospital，China Academy of Chinese Medical Sciences from January 2022 to September 2022 were included，and their bone marrow samples were collected by myelotomy. HIF-1α expression level in bone marrow cells was detected by real-time polymerase chain reaction （PCR） to analyze its correlation with clinical features，and logistic and Cox regression was used to analyze the risk factors affecting the efficacy and prognostic survival of MDS patients. ResultThe HIF-1α mRNA expression level was lower in bone marrow cells of MDS patients than in healthy subjects. HIF-1α was positively correlated with the degree of myelodysplasia（r=0.384，P<0.05） and bone marrow granulocytic system%（G%）（r=0.560，P<0.01）. Logistic regression showed that HIF-1α was a risk factor for the prognosis in the follow-up of the efficacy of treatment（P<0.05）and Cox regression showed that HIF-1α was an independent factor affecting the survival prognosis of MDS patients ［odds ratio（OR）=398.968，95% confidence interval（CI）（1.281，116 858.743），P<0.05］. ConclusionThe level of HIF-1α expression in bone marrow cells of MDS patients was closely related to the degree of clinical myelodysplasia and G%，and HIF-1α was a risk factor for the efficacy for and survival prognosis of MDS patients.

OBJECTIVETo observe the effect of Folium Panax quinquefolium saponins (PQS) on apoptosis of cardiac muscle cells (CMCs) and apoptosis-related gene expression in rats with acute myocardial infarction (AMI).

METHODSFifty Wistar rats were randomly divided into 5 groups, 40 rats underwent coronary ligation of left anterior descending branch to establish AMI rats model, while to the other 10 rats in the sham-operation group, thoracotomy without coronary ligation was carried out. The AMI model rats in treated groups were treated with high-dosage PQS, low-dosage PQS, captopril respectively for 7 days, and those in the control group were treated with normal saline instead. Then, the apoptotic CMCs were labeled by TUNEL and the expression of Bcl-2 and Fas in cardiac muscle cells were determined by immunohistochemical methods.

RESULTSThe apoptotic index of CMCs in the model rats (46.48%) was significantly higher than that in the sham-operation group ( 1.03%, P<0.01 ). CMCs apoptosis rate in the low-dosage PQS group, the large-dosage group and the captopril group was 23.53 %, 17.58 % and 25.17 %, respectively, all were significantly lower than that in the sham-operation group (P<0.01). The expression of Bcl-2 gene was up-regulated and expression of Fas protein was down-regulated in the three treated groups.

CONCLUSIONPQS can inhibit CMCs apoptosis in AMI rats, downregulate Fas protein expression and up-regulate Bcl-2 protein expression, and has antagonistic effect in myocardial ischemic injury.

Animals ; Apoptosis ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Male ; Myocardial Infarction ; blood ; genetics ; pathology ; Myocytes, Cardiac ; pathology ; Panax ; Plant Leaves ; chemistry ; Proteins ; genetics ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; biosynthesis ; genetics ; Random Allocation ; Rats ; Rats, Wistar ; Saponins ; pharmacology

OBJECTIVETo seek the key platelet functional proteins (PFPs) for the occurrence of blood-stasis pattern (BP) in patients with coronary heart disease (CHD).

METHODSPeripheral blood platelet protein of 22 patients and 24 healthy person (for control) were extracted respectively in 4 batches for carrying 4 times of the test out. Differential PFPs in samples were screened out by two-dimensional fluorescence difference gel electrophoresis, and identified with matrix-assisted laser desorption/ionization-time of flight mass spectrometry; then the identified proteins were further authenticated by Western-blotting.

RESULTSThirteen differential PFPs were screened out, and among them the 7 identified by spectrometry were: isoform 1 of integrin alpha- II b, isoform 2 of integrin alpha- II b, actin-cytoplasmic 1, actin-cytoplasmic 2, cDNA FLJ52842, cDNA FLJ55253, and cDNA FLJ43573 fis. Among them isoform 2 of integrin alpha- II b (CD41) and actin-cytoplasmic 2 (Acting) were authenticated successfully.

CONCLUSIONCD41 and acting are the possible marker proteins, and the other PFPs might play crucial roles in the occurrence and development of BSS in CHD.

Adult ; Aged ; Blood Platelets ; Case-Control Studies ; Coronary Disease ; blood ; diagnosis ; physiopathology ; Electrophoresis, Gel, Two-Dimensional ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Platelet Membrane Glycoproteins ; isolation & purification ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

OBJECTIVETo investigate the effects of Propyl Gallate (PrG) on serum inflammatory factors and protein expression of cyclooxygenase-2 (COX-2) and intercellular adhesion molecule-1 (ICAM-1) in ischemic myocardium of rats with acute myocardial infarction (AMI).

METHODSAMI model was induced by ligating the left anterior descending (LAD) branch of coronary artery in Wistar rats, and the perfect modeling was certified with ST segment elevation by standard limb lead II of electrocardiogram. Rats were randomly divided into 6 groups: Group A of normal rats, Group B of rats through sham operation, Group C of AMI model rats, Group D of model rats treated with high dose PrG (80 mg x kg(-1) x d(-1), via peritoneal injection), Group E of model rats treated with low dose PrG (40 mg x kg(-1) x d(-1), via peritoneal injection), and Group F of model rats treated with aspirin (25 mg x kg(-1) x d(-1), via gastrogavage), all the treatments were given in succession for 7 days. Radioimmunoassay (RIA) was used to determine serum contents of interleukin (IL)-1beta and tumor necrosis factor-alpha (TNF-alpha), immunohistochemistry was used to determine the level of COX-2 and ICAM-1 protein expression in myocardium.

RESULTSCompared with Group B, the serum level of TNF-alpha increased significantly, but not the level of IL-1beta in Group C. Besides, the COX-2 and ICAM-1 protein expressions in ischemic myocardium increased in Group C. All the above-mentioned changes were reversed to certain extent in Group E after treatment.

CONCLUSIONSPrG (40 mg x kg(-1) d(-1)) could decrease the serum level of inflammatory factor TNF-alpha, and slightly inhibit COX-2 and ICAM-1 protein expression in ischemic myocardium of AMI rats.

Animals ; Cyclooxygenase 2 ; genetics ; metabolism ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Gene Expression ; drug effects ; Humans ; Inflammation Mediators ; blood ; Intercellular Adhesion Molecule-1 ; genetics ; metabolism ; Male ; Myocardial Ischemia ; drug therapy ; genetics ; metabolism ; Myocardium ; metabolism ; Propyl Gallate ; administration & dosage ; Rats ; Rats, Wistar

The background, concept and status quo of translational medicine at home and abroad were introduced systematically in this review, and the application mode of translational medicine in the research and development of Chinese medicine (CM) was analyzed. Targeting the characteristics of CM and the changes in the spectrum of diseases in China, some suggestions were made to strengthen the translational research in CM and integrative medicine.
China ; Humans ; Integrative Medicine ; methods ; Translational Medical Research

OBJECTIVETo investigate the effect of Qidan Liquid (QL) on myocardial angiogenesis in rats after acute myocardial infarction (AMI) and explore its possible molecular mechanism.

METHODSAMI model was established by ligating the left anterior descending (LAD) coronary artery in Wistar rats, and intervened with QL in high, medium and low doses (QLh, QLm and QL1 group), and isosorbide dinitrate (ID group) respectively. Meanwhile, the model group, the sham group and the normal group were also set up. On the 7th (d7) and 14th day (d14) after modeling, mRNA and protein expressions of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) were detected with RT-PCR and immunohistochemistry, factor VIII expression in endothelial cells was examined, and microvascular density (MVD) in ischemic myocardium was calculated.

RESULTSMVD in the model rats increased significantly, higher than that in the normal and the sham group (P < 0.01), on d7 and d14, it was lower in the model group than that in the high and medium dose of QL groups and the ID group (P < 0.05). On d7, VEGF protein expression in the model group was higher than that in the normal group (P< 0.05), equivalent to that in the other groups, on d14, it was lower than that in the high and medium dose of QL groups and the ID group (P < 0.05 or P < 0.01). On d7 and d14, the protein expression of bFGF in the model rats increased significantly, higher than that in the normal and the sham group (P < 0.01), it decreased along the time progress between the two time points (P < 0.05), and lower than that in the high dose of QL group and ID group (P < 0.01). The mRNA expressions of VEGF and bFGF were higher in the model group than those in the normal and sham group, but lower than those in all the treated groups on d7 and d14. Comparison of the strips on d7 and d14 among various groups showed that the bFGF mRNA expression in the model group showed a descending trend as the time goes on, but in all the treated groups it was unchanged, while the change of VEGF mRNA expression didn't show any definite tendency.

CONCLUSIONQL could up-regulate the protein and gene expressions of VEGF and bFGF persistently and increase MVD in ischemic myocardium to promote the development of collateral circulation in AMI rats.

Angiogenesis Inducing Agents ; therapeutic use ; Animals ; Coronary Circulation ; drug effects ; Drugs, Chinese Herbal ; therapeutic use ; Male ; Myocardial Infarction ; drug therapy ; Neovascularization, Physiologic ; drug effects ; Phytotherapy ; Rats ; Rats, Wistar ; Solutions