Background Eicosapentaenoic acid(EPA)function as the critical lipid mediators involved in several biological events in human body and play important role in suppressing the genesis of vascular endothelial growth factor (VEGF),migration and proliferation of vascular endothelial cells.Many ocular diseases were proved to be associated with neovascularization.Objecfive The purpose of this study was to investigate the inhibitory effect of EPA on the proliferation of human umbilical vein endothelial cells (HUVEC) indueed by VEGF. Methods HUVEC strain was cultured and passaged,and difierent concentrations of EPA were added to the medium with and without VEGF.The cultured cells were identified by antiofactor Ⅷ polyclonal antibody.The suppressing role of different concentrations of EPA on the proliferation of VEGF-induced or-uninduced HUVEC was assessed by MTT method.The influence of difierent concentrations of EPA on the cellular cycle of VEGF-induced HUVEC was assayed using flow eytometry.The expression of Flk-1,a receptor of VEGF,in the HUVEC Was detected by immunohistochemistry. Results Cultured HUVEC showed the ftlsiform in shape and presented with the cobblestone-like arrangement with the positive response for Ⅷ factor-related antigen.Various concentrations of EPA showed obviously inhibitory effect on VEGF-induced or-unindueed HUVEC at a dose-dependent manner (F=23.072.P=0.000).The inhibitory ability of EPA on VEGF-induced HUVEC was stronger than VEGF-uninduced HUVEC(F=41.417,P=0.000).In 24,48 and 72 hours,the action of EPA on the proliferation of HUVEC was gradually enhanced with the prolong of time(F=1.495,P=0.236).Cell cycle analysis indicated that EPA arrested VEGF-induced HUVEC in G0/G1 phase.The ratio of HUVEC in G0/G1 phase in EPA group was(75.83±1.56)%,and that in control groups was(68.62±1.44)%,showing a significant difference between them(t=-5.88,P=0.00),and no apoptosis of HUVEC was found in both groups.Flk-1 was strongly expressed in the cellular nucleus and cytoplasm in control group.However,the positive expressing intensity of Flk-1 in the HUVEC weakened,and the positive cell number was evidently less in EPA group. Conclusion EPA can inhibit the proliferation of VEGF induced HUVEC through arresting the synthesis of DNA of HUVEC and downregulate the expression of Flk-1 in HUVEC.These results suggest that EPA might exert an antiangiogenic effect.

Objective To establish an simple and convenient animal model of cardiac arrest for studying cerebral resuscitation.Method Clean male Sprague Dawley rats were randomly divided into sham group and experimental group.Cardiac arrest was induced by asphyxiation and ice-cold 0.5 mol KCl with blood flow cut off for 5 minutes.Animals were resuscitated with external cardiopulmonary resuscitation (CPR),mechanical ventilation,and epinephrine injection.Blood pressure,heart rate,successful ratio of resuscitation after 72 hours, time of cardiac arrest (T_(CA)) and return of spontaneous circulation (T_(ROSC)) were recorded.Neural deficit scores (NDS) and levels of maleic dialdehyde (MDA) in plasma were evaluated at 3,6,12,24,48,72 hours after ROSC.The damage score of cortex was measured by transmission electron microscope examination at 3 hours and 72 hours after ROSC.Results All the rats in experimental group had cardiac arrest rapidly.T_(CA) and T_(ROSC) were (137.3?10.2) seconds and (64.4?9.3) seconds,respectively,while the successful rate of resuscitation was 82.5%.The lowest NDS was at 3 hours after ROSC,while the NDS increased gradually.After CPR,the level of MDA in plasma increased significantly,slightly declined at 72 hours after ROSC,but still significantly higher than before the model.Electron microscope examination of cortex showed neuron slightly,organelle and astrocyte,but became better after 72 hours post ROSC.Conclusions The model of cardiac arrest was easy to establish,and the data provided was accurate,which is useful to study the mechanism of cerebral resuscitation.

Objective: To analyze and compare the features and strengths of different methods for calculating the biophysical properties of meridian points, and thus propose corresponding suggestions to fully achieve the research and application value of biophysical properties of meridian points.Methods: We searched and collected the literature on the imbalance of biophysical properties of meridian points between January 1, 2005 and March 1, 2020 in China National Knowledge Infrastructure (CNKI), Wanfang Academic Journal Full-text Database (Wanfang), Chongqing VIP Database (CQVIP) and PubMed database, and then analyzed, compared, and summarized the applied methods for calculating the imbalance degree of the biophysical properties of meridian points. Results: The current methods for calculating the imbalance degree of the biophysical properties of meridian points are diverse and can be summarized as the following three: direct comparison of the measured values of the left and right namesake points, difference method, and ratio method. The low uniformity of the calculation methods has limited the promotion and application of its research results. Conclusion: In future research on the biophysical properties of meridian points, multidisciplinary cooperation in terms of imbalance degree calculation methods, detection instruments, and health data models is necessary to achieve more widely applicable scientific conclusions and more generalized experimental results.

OBJECTIVETo investigate the risk factors of female breast cancer and its potential alteration in Wuhan area.

METHODSA case-control study was conducted on 213 cases with histopathological diagnosis and 430 matched controls, using conditional logistic regression analysis.

RESULTS28 factors such as educational level, history of benign breast disease, age at menarche, age at menopausal, meat and well-done meat intake, soy bean food, fruit, lactation time, body mass index (BMI), juvenile chest X-ray, psychological factor, were associated with breast cancer risk in one-way variance model. Multivariate conditional logistic regression analysis in total significant factors and subgroups showed that the risk factors of breast cancer would include high level education, psychological trauma, history of benign breast disease later age at menopause, more years of menstrual and more years of menstrual before giving first birth, high BMI, well-done meat intake and smoked food. Factors as later menarche, lactate longer, soybean food, fruit, drink tea habit were protective factors for breast cancer. Further breakdown of data showed some difference between premenopausal and postmenopausal women. Risks in premenopausal women were associated with history of benign breast disease, age of menarche, soybean food intake, whereas risks in postmenopausal women were related to age of menopausal, BMI, waist-hip ratio and fruit intake. Both psychological traumatic and duration of lactation were common pre-and postmenopausal risk and protective factors.

CONCLUSIONSDietary habit and endogenous estrogen exposure related factors played important roles on women breast cancer in Wuhan area.

Adult ; Breast Neoplasms ; etiology ; metabolism ; Case-Control Studies ; China ; Estradiol Congeners ; metabolism ; Feeding Behavior ; Female ; Humans ; Logistic Models ; Middle Aged ; Multivariate Analysis ; Risk Factors

AIMThe present study was designed to determined the cardiovascular effects of IMD1-53 in rats and its possible mechanism.

METHODSIsolated rat hearts were perfused by Iangendorff mode, and ventricular function was measured after IMD1-53 perfusion. Meanwhere, we investigated the effects of IMDI) on arterial pressure after intravenous administration of IMD. And cAMP content was detected in rat ventricular and aortic tissues.

RESULTSThe results showed that perfusion with IMD significantly enhanced cardiac function and resulted in higher LVSP, +dp/dt(max) and -dp/dt(max) by 45%, 51% and 37%, respectively, compared with control and increased coronary infusion flow. The effects of IMD1-53 on cardiac function were antagonized by H-89, an inhibitor of PKA. The content of cAMP in the ventricular tissues after IMD perfusion was 131% higher than control. In addition, intravenous administration of IMD induced a potent decrease in arterial pressureand heart rate, and in aortic tissues, IMD incubation resulted in a 236% increase in cAMP content compared with control group.

CONCLUSIONThe study reveals that IMD can increase cardiac function and decrease arterial pressure in rat and the effects may be related to cAMP pathway.

Adrenomedullin ; metabolism ; pharmacology ; Animals ; Blood Pressure ; drug effects ; Cardiovascular Physiological Phenomena ; drug effects ; Cyclic AMP ; metabolism ; Heart ; drug effects ; In Vitro Techniques ; Male ; Neuropeptides ; metabolism ; pharmacology ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Ventricular Function ; drug effects

The aim of this study is to investigate the critical factor affecting the properties of PLGA microspheres fabricated by a solid-in-oil-in-water (S/O/W) emulsion technique with BSA as a model protein. Prior to encapsulation, the BSA microparticles were fabricated by a modified freezing-induced phase separation method. The microparticles were subsequently encapsulated into PLGA microspheres by S/O/W emulsion method, then Motic BA200 biological microscope, confocal laser scanning microscope, scanning electron microscope were used to observe the structure of S/O/W emulsion and PLGA microspheres. The protein content extracted or released from BSA microspheres was measured by Bradford protein assay method. It was found that NaCl added in the outer aqueous phase effectively suppressed material exchange between the inner and outer phase of S/O/W emulsion. Then, the structure and permeability of obtained microspheres were influenced. As a result, with the increase of NaCl concentration in the outer aqueous phase, the encapsulation efficiency of microspheres significantly increased from 60% to more than 85%, the burst release of microspheres reduced from 70% to 20%, and the particle size decreased from 103 microm to 62 microm. Furthermore, the rehydration of encapsulated protein was also retarded and then integrity of BSA was successfully protected during encapsulation process. In vitro release test showed that BSA released from PLGA microspheres in a sustained manner for more than 30 days.
Delayed-Action Preparations ; Drug Compounding ; Emulsions ; chemistry ; Lactic Acid ; administration & dosage ; chemistry ; Microscopy, Confocal ; Microscopy, Electron, Scanning ; Microspheres ; Oils ; Particle Size ; Polyglycolic Acid ; administration & dosage ; chemistry ; Serum Albumin, Bovine ; administration & dosage ; chemistry ; Sodium Chloride ; chemistry ; Water

OBJECTIVETo construct the eukaryotic expression vector PCI-dhfr-lambda1 and PCI-dhfr-SP163-lambda1 which linked the enhancer SP163 with interferon lambda1. Then express the interferon lambda1 in CHO (dhfr-) cells.

METHODSUsing PCR method to introduce the restriction enzyme sites and through the fusion PCR binding the enhancer with the interferon Lambda1. After sequenced, lambda1 and SP163-lambda1 was inserted into PCI-dhfr forming the expression vector PCI-dhfr-lambda1 and PCI-dhfr-SP163-lambda1 which was constructed successfully confirming by sequencing. Then the expressing vectors were transfected into CHO (dhfr-) cells using liposome transfection method and interferon lambda1 protein was assayed with indirect immunofluorescence and Western Blot. Using cytopathic effect inhibition evaluated the antiviral activity of interferon lambda1.

RESULTSSuccessfully constructing the eukaryotic expression vectors of interferon lambda and the vectors could express interferon lambda1. The result of immunofluorescence showed the enhancer developed the expression of interferon lambda1. Detecting the interferon lambda1 in CHO (dhfr-) cells after transfecting 48 hour using Western Blot. The cytopathic effect inhibition showed the expressed interferon lambda1 has the antiviral activity.

CONCLUSIONSuccessfully expressed the interferon lambda1 in CHO (dhfr-) cells and the protein possesses antiviral activity, which may supply a valuable basis for building the stable cell line of interferon lambda1.

Animals ; Blotting, Western ; CHO Cells ; Cricetinae ; Cricetulus ; Fluorescent Antibody Technique, Indirect ; Interleukins ; genetics ; pharmacology ; Polymerase Chain Reaction ; Recombinant Proteins ; biosynthesis ; pharmacology ; Transfection

OBJECTIVETo report a case of acute myeloid leukemia (AML) with the insertion (8;21)(q22;q22.1q22.3). A 33-year-old Chinese woman was referred to our hospital. Hematologic data showed WBC 42.7 x 10(9)/L with monocytosis (monocyte counts 7.296 x 10(9)/L). Bone marrow aspirate was hypercellular with 4.5% monoblasts and 7.5% promonocytes. At first she was diagnosed with chronic myelomonocytic leukemia (CMML) according to the FAB criteria. Initially the patient received supportive care only, but her general condition rapidly became worse three months later. The monoblasts and promonocytes in the bone marrow rose to 20.5%. After two cycles of combined chemotherapy she obtained complete remission.

METHODSChromosome specimens were prepared by short-term culture of bone marrow cells. Karyotype analysis was carried out by R-banding technique. Three fluorescence in situ hybridization (FISH) analyses were performed using AML1-ETO dual color, dual fusion probe, whole chromosome painting 8 and 21 probes, and cen-8 and Tel 21qter probes, respectively. Reverse transcription polymerase chain reaction (RT-PCR) assay for detecting the AML1-ETO fusion transcript was also performed.

RESULTSConventional cytogenetic analysis showed a karyotype of 46,XX,ins(8;21) (q22;q22.1q22.3)[7]/46,XX[3]. FISH tests confirmed the insertion. RT-PCR analysis detected the AML1-ETO fusion transcript.

CONCLUSIONWe consider that this patient should be rediagnosed as acute myeloid leukemia according to the criteria proposed by World Health Organization (WHO) and that FISH and RT-PCR play an important role in verification of the ins(8;21).

Chromosome Banding ; Chromosomes, Human, Pair 15 ; Chromosomes, Human, Pair 19 ; Chromosomes, Human, Pair 8 ; Core Binding Factor Alpha 2 Subunit ; genetics ; Female ; Humans ; In Situ Hybridization, Fluorescence ; methods ; Karyotyping ; Leukemia, Myeloid ; genetics ; Translocation, Genetic

OBJECTIVETo study CXCR3 and CCR5 chemokine receptor expression in spleens of patients with primary immune thrombocytopenia (ITP) and its clinical significance.

METHODSThe splenectomy specimens from 10 ITP patients (ITP group) and 8 patients with traumatic splenic rupture (normal control group) were studied. Immunohistochemistry (IHC) was used to study the positive rate of CXCR3 and CCR5. Western blot was performed to detect CXCR3 and CCR5 protein expression, while real-time polymerase chain reaction (RT-PCR) was conducted to analyze their mRNA expression.

RESULTSThe positive rate of CXCR3 and CCR5 were both higher in ITP group (90% and 100%, respectively) than those in control group (75% and 87.5%, respectively)(P < 0.05). The differences were statistically significant (P < 0.05). Protein and mRNA level of CXCR3 in ITP group were 3.0 and 3.5 times as high as those in control group, respectively. Those of CCR5 in ITP group were 1.2 and 1.7 times as high as those in control group, respectively.

CONCLUSIONHigh expression of CXCR3 and CCR5 may play a part in the splenic immune disorders in patients with ITP.

Adolescent ; Adult ; Case-Control Studies ; Child ; Female ; Humans ; Male ; Middle Aged ; Receptors, CCR5 ; metabolism ; Receptors, CXCR3 ; metabolism ; Spleen ; metabolism ; Thrombocytopenia ; immunology ; metabolism ; Young Adult

OBJECTIVETo express and purify HBoV VP2 protein, and the monoclonal antibody against HBoV VP2 protein was prepared with hybridoma technique.

METHODSThe HBoV VP2 cloned into vector pET-30a was expressed in E. coil. After purified by immobilized metal affinity chromatography, the BALB/c mouse was immunized with purified protein as antigen. The positive hybridoma cells were screened with hybridoma technique and ELISA assay. Isotype and titer of the monoclonal antibody were detected.

RESULTSThe recombinant HBoV VP2 protein was expressed and purified, and then the monoclonal antibody was obtained with hybridoma technique. The titer of the IgG monoclonal antibody was up to 1:4 x 10(5).

CONCLUSIONMonoclonal antibody against recombinant HBoV VP2 protein was prepared and the antibody titer was high. This work may provide a new method in rapid diagnosis and study of HBoV.

Animals ; Antibodies, Monoclonal ; immunology ; Capsid Proteins ; genetics ; immunology ; Human bocavirus ; immunology ; Hybridomas ; Mice ; Mice, Inbred BALB C ; Plasmids ; Recombinant Proteins ; biosynthesis ; immunology ; isolation & purification