Objective To study the expression of PTEN and Caspase-3 proteins in esophageal squamous cell carcinoma(ESCC) and to analyze the relationships between the expressions of the proteins and clinicopathologic factors as well as prognosis.Methods 64 cases of ESCCs,16 cases of pericarcinoma tissues and 16 cases of dysplasia epithelium were immunohistochemically stained.All 64 patients who underwent surgical treatment up to January 2005 were regularly followed up through letters and check-ups.Results The positive expression rate in pericarcinoma tissues,dysplasia and ESCC were significantly down-regulated((P

BACKGROUND:At present, a great quantity of research has shown the effectiveness of traditional Chinese medicine and bone marrow mesenchymal stem cel s for vascular restenosis. However, studies concerning their combined application to restenosis after percutaneous transluminal angioplasty with diabetes mel itus are presently lacking.
OBJECTIVE:To observe the effects of combined application of bone marrow mesenchymal stem cel s and benefiting-Qi nourishing-Yin and dissolving-congestion prescription on restenosis after percutaneous transluminal angioplasty in dogs with diabetes mel itus.
METHODS:A dog model of vascular restenosis with diabetes mel itus was established by bal oon injury of femoral artery and intravenous injection of al oxan. After successful model induction, 22 dog models were randomly divided into three groups:model group (n=6), treatment with Chinese medicine (n=8), and combined treatment with bone marrow mesenchymal stem cel s and Chinese medicine (n=8). Serum vascular endothelial growth factor levels were measured using enzyme-linked immunosorbent assay preoperatively and at 1, 2, 4 and 8 weeks postoperation. Samples of vessels were taken to conduct pathomorphological observation and quantitative analysis of proliferation degree. Tissues, including heart, liver, kidney and pancreatic gland, were col ected to evaluate the safety of stem cel transplantation using hematoxylin-eosin staining at 8 weeks postoperation.
RESULTS AND CONCLUSION:Serum vascular endothelial growth factor levels began to increase at 1 week postoperation in the Chinese medicine group and combined treatment group, at 4 weeks postoperation in the model group compared with preoperation (P<0.05). At al time points, serum vascular endothelial growth factor levels were highest in the combined treatment group, but lowest in the model group (P<0.05). Quantitative analysis of vascular proliferation demonstrated that at 8 weeks postoperation, new intimal area, new intimal/medial areas and stenosis rate were highest in the model group, but lowest in the combined treatment group at 8 weeks postoperation (P<0.05). Safety assessment of stem cel transplantation showed morphological structures of the heart, liver, kidneys and pancreas were normal, no necrosis. In a word, the effects of the combined application of bone marrow mesenchymal stem cel s and benefiting-Qi nourishing-Yin and dissolving-congestion prescription were much pronounced in preventing restenosis after percutaneous transluminal angioplasty in dogs with diabetes mel itus rather than single therapy of Chinese medicine. It is a safe and effective treatment to prevent vascular restenosis after percutaneous transluminal angioplasty in dogs with diabetes mel itus.

OBJECTIVETo study the inhibition of berberine (BBR) against ECV-304 apoptosis induced by Staphylococcus aureus (S. aureus).

METHODSECV-304 cells were pre-treated with 128 microg/mL BBR for 2 h and then S. aureus was added (1:100). The viability of cells was detected by MTT (3-4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. The morphological changes were observed by Hoechst 33258 staining. The protection of BBR for infected cells was detected by DNA Ladder.

RESULTSECV-304 cells' viability were not obviously affected by berberine. But S. aureus induced ECV-304 cells' viability could be significantly inhibited by pre-treatment of BBR (P < 0.05). Besides S. aureus-induced ECV-304 apoptosis could be reduced, with significantly lessened apoptotic body and unobvious DNA degradation.

CONCLUSIONBBR could significantly inhibit S. aureus induced ECV-304 apoptosis.

Apoptosis ; drug effects ; Berberine ; pharmacology ; Cell Line ; Human Umbilical Vein Endothelial Cells ; drug effects ; microbiology ; pathology ; Humans ; Staphylococcus aureus

Objective To obtain antibiotic-resistant mutants producing metabolites with antitumor activity from wild-type actinomycete strains without antitumor activity. Methods An actinomycete strain L35-1 was used as an initial strain for obtaining antibiotic-resistant mutants, which is a marine-derived wild-type strain without antitumor activity with an inhibition rate of 2.8% at the 1000 μg/ml of high sample concentration on K562 cells. The antibiotic-resistant mutants both from auto-mutagenesis and chemical mutagen-induced mutagenesis were selected by single colony isolation on antibiotic-containing plates according to the method for obtaining drug-resistant mutants in ribosome engineering. The antitumor activity was assayed by the MTT method using K562 cells for the mutants with aqueous acetone extracts of the whole broth of their fermentation.Results A total of 114 neomycin-resistant (ner) and 68 streptomycin-resistant (str) mutants, all from auto-mutagenesis, was obtained on drug-containing plates. Among them, the 7 ner and 3 str mutants appeared to be bioactive with an inhibition rate above 20% at the 100 μg/ml sample concentration on K562 cells. On the other hand, 41 str and 32 ner mutants from DES-induced mutagenesis and 46 ner mutants from NTG-induced mutagenesis were obtained by mutagen-induced mutation coupled with the single colony isolation on antibiotic-containing plates, among which, one str mutant from DES-induced mutagenesis and one ner mutant from NTG-induced mutagenesis were bioactive with an inhibition rate over 20% at the 100 μg/ml sample concentration on K562 cells. Conclusions The present result has revealed that the wild-type actinomycete strains without bioactivity might become a great source initial strains to obtain bioactive mutants by drug-resistant mutation technique.

Objective To study influencing factors for giving up chemotherapy during initial treatment in elderly patients with multiple myeloma.Methods A total of 156 elderly patients diagnosed as multiple myeloma from 2000-2010 were retrospectively analyzed.And 123 young patients with multiple myeloma were selected as control group.The causes for giving up the chemotherapy and its related factors were recorded and analyzed.Results The rate of giving up the initial chemotherapy was higher in the elderly than in young patients [58.3％ (91/156) vs.35.0％ (43/123),P＜0.05].51.7％ of the elderly patients did not accept chemotherapy after diagnosis.The main factors for giving up the initial chemotherapy in elderly patients were critically ill,economic difficult and the low tolerance.However,the economic difficulty was the first cause in the young group.Conclusions The rate of giving up the chemotherapy in the elderly patients is high.Old age,severe conditions,economic difficulty,and low tolerance are the independent factors.

Objective To examine the expression of TLR7/8 in monocytes purified from HIV-1 infected individuals and to study its association with disease progression. Methods Sixty-three HIV-1 infected individuals and 18 normal controls were enrolled. Monocytes were purified by MACS system and RNA of them was extracted by RNA mini kit of QIAGEN company. TLR7/8 expression was tested by real-time RT-PCR with ABI7500. Results It was found that the expression of TLR7 was strongly correlated with absolute CD4 count (r =0.614, P＜0.01) , so was TLR8 (r =0.419, P＜0.01). The expression of TLR7 in slow progressor (SP) group was higher than that in HIV-1 infected patients group, AIDS patients group and normal group (P ＜ 0. 05 ) . HIV group and normal group were strongly higher than AIDS group (P ＜ 0. 05). It was no significant differentiation of expression of TLR7 between HIV infection group and normal control group. The expression of TLR8 in SP group and normal group were significantly higher than that in AIDS group (P ＜ 0. 05). The expression of TLR8 was no singnificantly difference between SP group and HIV group or normal control group, so was it between HIV group or normal control group and AIDS group. Conclusion The expression of TLR7/8 in monocytes from HIV-1 infected patients significantly correlated with disease progression.

Background The molecular biological mechanisms of diabetic retinopathy(DR)is unclear up to now.Researches have proved that endoplasmic reticulum stress(ER Stress)-associated factors are elevated in peripheral blood in patients with diabetic retinopathy.and P58 IPK can inhibit those factors.So the relationship between P58 IPK and DR is worth to investigate. Objective The aim of this study was to detect the dynamic expression of P58 IPK in the retina of diabetic rats. Methods The diabetic animal models were established in 18 clean male SD rats by intraperitoneal injection of stilptozotiein(STZ)at a dose of 60 mg/kg.The rats were sacrificed in 1,3,6 months after injection.The expression change of P58 IPK mRNA in the rats retina was detected by quantitative real-time RT-PCR.Other 6 matched normal rats were used as control groups.This experiment followed the Regulations for the Administration of Affair Concerning experimental Animals by State Science and Technology Commission. Results The rats showed more drinking,more food and more urine after STZ injection with the blood glucose level≥ 16.5 mmol/L.The success rate of diabetic models was 100%.The A value of P58 IPK mRNA/β-actin in rat retina was 0.800±0.005 and 0.975±0.008 after injection of STZ.and that of control rats was 0.725±0.006,showing statistically significant difference between them(t=22.589,t=62.784,P＜0.05).In 6 months after injection of STZ,the expression of P58 IPK mRNA in experimental diabetic rat retina was evidently lower than the eontrol rats(0.671±0.004 versus 0.725±0.006,t=-17.984,P＜0.05).Conclusion P58 IPK has a close relation to the pathogenesis of DR,and it plays a retarding role for DR.

Background The in vitro culture of retinal vascular endothelial cells is the foundation of experimental study of retinal vascular disease. Shortage of human donor eyeballs is a main limiting for the laboratory work. The culture method of rat-derived vascular endothelial cells has been established. However, this method is not enough effective because of severer cellullar injury. Objective Present study was to establish a simple and high effective method for the culture of vascular endothelial cells in vitro. Methods The retinas from 5 SPF SD rats was digested by 0. 1% collagenase and cultured with explant culture method. 20% fetal bovine serum, vascular endothelial growth factor ( VEGF) , insulin-transferrin-selenium( ITS) were composed into the endothelial cell culture medium, and enough blowing was performed to get the cells and fragments from retinal tissue. The cellular suspension was prepared and cultured consequently on human fibronectin-coated culture flasks. Cultured vascular endothelial cells were identified by anti-von Willebrand staining factor. Results The cells emerged from the tissue mass,and cells and some tissue fragments attached to the wall after 24 hours of seeding. The cells grew to show the fusiform in 4 days and merged together in 5 to 6 days,and a cell monolayer was seen in the 14th day after culture. The endothelial cells showed the positive response for von Willebrand factor. After passage, the merging-growth statue of the cells was regained in 2 hours after culture. Conclusion Use of retinal pieces and collagenase-digestion can get the vascular endothelial cells with better activity in vitro. The culture method based on highly selective endothelial cell culture medium associated to FN adhesion-promoting is helpful for gaining the purified of endothelial cells.

Objective:To explore whether there were changes in midbrain morphology related to motor function in patients with cerebral small vessel disease (CSVD).Methods:The study was conducted on a subset of patients with CSVD registered at the Department of Neurology of Peking Union Medical College Hospital from 2010 to 2018. All magnetic resonance imaging images were taken with a 3.0 T nuclear magnetic resonance imager. The measurement of anteroposterior diameter of the mesencephalon and the tegmentum of mesencephalon was performed on a personal computer with the image processing software RadiAnt DICOM Viewer. Several clinical manifestations of dyspraxia, dysphagia, dysarthria and dysuria were evaluated by interviewing the patient and family members living with the patient. Kinect depth camera combined with self-developed software platform was used to conduct quantitative evaluation of patients′ motor function. Three parameters, namely walking speed, walking time of three meters and time of standing up and sitting down, were selected from the obtained parameters. SPSS 17.0 software was used for statistical analysis.Results:A total of 176 patients were included, aged 30-88 (64.16±11.57) years. One hundred and fifteen patients were males, accounting for 65.34%. In patients with CSVD, anteroposterior diameter of the mesencephalon and the tegmentum of mesencephalon were negatively correlated with age ( B=-0.032, P<0.001; B=-0.020, P=0.006). The anteroposterior diameter of the mesencephalon was negatively correlated with symptoms of dyskinesia ( OR=0.006, 95% CI 0-0.135, P=0.001), even when the age and gender were adjusted ( OR=0.014,95% CI 0-0.416, P=0.013). The anteroposterior diameter of the mesencephalon was negatively correlated with symptoms of dysuria ( OR=0.046,95% CI 0.002-0.936, P=0.045), but no longer correlated when the age and gender were adjusted. The anteroposterior diameter of tegmentum of mesencephalo was also negatively correlated with symptoms of dyskinesia ( OR=0.035,95% CI 0.002-0.684, P=0.027), but no longer correlated after adjusting for age and gender. Video recording and evaluation of motor function were performed on 87 patients. Spearman correlation analysis showed that none of the three motor function scores was correlated with age or gender. The anteroposteric diameter of the midbrain was positively correlated with walking speed ( r=0.231 ,P=0.040) and negatively correlated with walking time of three meters ( r=-0.304, P=0.005), but not with standing up and sitting down time, while the anteroposteric diameter of tegmentum of mesencephalo was not correlated with all of them. Conclusion:There are changes in mesencephalon morphology in patients with CSVD, structural parameters of mesencephalon are related to motor function and urination function, and mesencephalon atrophy may be an independent related factor for symptoms of dyskinesia in patients with CSVD.

OBJECTIVETo explore myocardial perfusion of patients with coronary slow flow (CSF) using myocardial contrast echocardiography.

METHODSMyocardial contrast echocardiography was performed in coronary artery angiography diagnosed CSF patients (n = 20) and control patients (n = 20). The images at baseline and after low dose dobutamine stress test were analyzed by automatic tracking software and the maximal amplitude score A, the mean ascending slope of the curve β and the product of A×β were measured. The reserve of A×β was also calculated. Electrocardiogram at rest and at each stage of dobutamine stress test was obtained simultaneously.

RESULTSAt baseline, the A [(6.85 ± 2.99) dB vs. (7.01 ± 3.49) dB], β[(0.59 ± 0.33)/s vs. (0.61 ± 0.38)/s] and A×β [(3.48 ± 1.46) dB/s vs. (3.31 ± 0.96) dB/s] values were similar between CSF group and control group (all P > 0.05). After dobutamine stress test, both β and A×β were significantly increased in two groups. The β [(0.89 ± 0.42)/s vs. (1.31 ± 0.54)/s, P < 0.01] and A×β [(5.82 ± 2.69) dB/s vs. (8.07 ± 2.76)dB/s, P < 0.05] in CSF group were significantly lower than in control group. Electrocardiogram of all the subjects was normal at rest, but the electrocardiogram positive rate was significantly higher in CSF group than in control group after dobutamine stress test (12% vs. 2%, 60% vs. 10%, P < 0.01).

CONCLUSIONSDobutamine stress test could induce myocardial perfusion abnormalities in patients with coronary slow flow phenomenon.

Aged ; Case-Control Studies ; Coronary Circulation ; drug effects ; Dobutamine ; Echocardiography ; methods ; Exercise Test ; Female ; Humans ; Male ; Middle Aged