BACKGROUND:Autologous conditioned serum (ACS) is a new and safe biological therapy method mainly used for osteoarthritis, rheumatoid arthritis and other similar diseases. OBJECTIVE:To explore the purification method and cytokine profile of ACS. METHODS:Blood obtained equipment (containing 3 mm of glass beads) was used to collect peripheral blood samples that were mixed uniformly and then placed in a 37℃ thermostatic incubator for 24 hours, finally filtered to obtain ACS. The quantitative determination of interleukin (IL)-1Ra, IL-6, IL-8, and IL-1β was conducted, and the 36 kinds of cytokines were semi-quantitatively detected by cytokine antibody array. RESULTS AND CONCLUSION: The levels of various cytokines in ACS were obviously increased, among which, IL-1Ra level (important for alleviating osteoarthritis) was increased from 140.25 ng/L to 1125 ng/L(increased by above 8 times); IL-6 level was increased from 389.5 ng/L to 2802 ng/L; IL-8 level was not detected firstly and finally increased to 2822 ng/L. Cytokine antibody array results showed that at least 10 kinds of cytokines in ACS were changed, and most of cytokines were on a rise, and few in decline. These results indicate that the ACS contains IL-1Ra and a variety of cytokines, and some chemokines in the ACS recruit more immune cells to the inflammation sites and accelerate the inflammatory process, further exerting therapeutic effects.

Objective To investigate the regulatory effects of icariin (ICA) combined with the Panax notoginseng saponins (PNS) on immunological function in mice and provide some experimental evidences for the combination mechanism improving the spatial learning and memory abilities of Alzheimer's disease (AD) animal model. Methods Based on serum pharmacological method, the ICR mice were individually ig administrated with ICA+PNS [(40+320), (80+640), and (160+ 1 280 ) mg/kg] or ICA (80 mg/kg), and PNS (640 mg/kg) only for 7 d. Drug-containing serum was prepared and effects on spleen lymphocyte proliferation of Bable/c mice induced by concanavalin-A (ConA) or lipopolysaccharide (LPS) and on interleukin-2 (IL-2) secretion were observed in vitro. Meanwhile, the immunological organ indexes of treated mice were evaluated. Results Drug-containing serum of ICA+PNS [(80+640) and (160+ 1 280 ) mg/kg] could improve the spleen lymphocyte proliferation induced by ConA (P0.05). Drug-containing serum of ICA+PNS [(80+640) and (160+ 1 280) mg/kg] could improve IL-2 production (P0.05), respectively. Conclusion ICA combined with PNS could improve immunological function selectively and promote T cell function in mice.

Objective To study the antigenicity of RNA polymerase and the relationship between the protein and the SARS-CoV replication. Methods The fragment was amplified by PCR, ligated with the prokaryotic expression vector pET32a+ and transformed into E. coli BL-21 .The expressed fusion protein identified by Western-blot and ELISA was used to detect the anti-SARS CoV IgG in different sera. Results The fusion protein was expressed successfully in E. coli BL-21. Detected by ELISA, the positive percentage of the anti-SARS IgG in the healthy donors, patients, infected animals and the rhesus administrated with the inactivated-virus were 0%, 95%, 100% and 0%, respectively. Conclusion Good antigenicity was shown in the expressed RNA polymerase. It can be used to diagnose the infection and to demonstrate the replication of SARS-CoV virus.

Objective To observe the therapeutic efficacy and safety of the glucagon-like peptide-1 ( GLP-1 )analogue combined with isulin for treating obese or overweight type 2 diabetes.Methods 40 cases with obese or overweight type 2 diabetic patients( body mass index(BMI) ≥24) who have previously used human insulin,and experienced poor glycemic control( ≥7.5％ ) were selected.They were randomly divided into two groups.A group treated by GLP-1 analogue liraglutide plus insulin,and B group by continuous adjustment of insulin.12 weeks later,fasting blood glucose(FBG),2-hour plasma glucose (2hPG),hemoglobin A 1 C (HbA1c),body weight were observed and the incidence of hypoglycemia,insulin dosage were recorded.Results Weight and insulin dosage of group A was significantly lower than those of group B after treatment 12 week( t =2.738,2.865,all P ＜ 0.01 ).Numbers of hypoglycemia events were 6 in group A(30％ ),and 9 in group B(45％ ),but there was no statistical significance between the two groups ( P ＞ 0.05).Conclusion The addition of liraglutide to insulin in patients with obese or overweight type 2 diabetes is associated with reductions in weight and insulin dosage,without increase risk of hypoglycemia.This treatment proved effective and safe.

Objective To investigate the effects of methylthioadenosine phosphorylase (MTAP) on invasion and migration in breast cancer cells.Methods Human breast cancer cell line MCF-7 cells were treated with MTAPtargeted siRNA to diminish MTAP mRNA.MCF-7 cells proliferation was evaluated by cell counting kit-8,the analysis of cells invasion and migration was performed using Transwell chamber.The expressions of MTAP and matrix metalloproteinase 1 (MMP1) in cell extracts were detected by Western blotting.The experimental divided into blank contrd group,negative control group,MTAP-siRNA experimental group.Results The MCF-7 cells growth was promoted after knockdown the MTAP mRNA.MATP-siRNA experimental group 450 nm absorbance values at 24h,48 hand72 h of the control group were (112.3±11.9)％,(144.4±8.4)％,(169.3±9.4)％ respectively.Cell invasion analysis by Transwell chamber showed 570 nm absorbance values were 0.49 ± 0.06 (control),0.45 ± 0.07 (negative control) and 0.87 ± 0.07 (MTAP-siRNA) respectively.Cell migration analysis by Transwell chamber showed 570 nm absorbance values were 0.46 ± 0.06 (control),0.49 ± 0.08 (negative control)and 0.75 ± 0.07 (MTAP-siRNA) respectively.The expression of MMP1 in MCF-7 cells was upregulated after knockdown the MTAP mRNA.Conclusion The knockdown of MTAP in MCF-7 cell can increase the cells invasion and migration,and this may involve the the MMP1.

Objective To investigate the effects of proanthocyanidins on depressant-like behaviors and the structure of adrenal gland in chronic unpredictable mild stress (CUMS) rats. Methods Rats were randomly divided into 6 groups:control group, stressed group (CUMS + vehicle), three treatment groups (CUMS + proanthocyanidins 25,50,100 mg·kg-1,respectively) ,and imipramine group (CUMS + imipramine 10 mg·kg-1). Used the CUMS model in rats to investigate the effects of chronic oral administration (21 days) of proanthocyanidins and imipramine (ip) on the open-field;and forced swimming and sucrose consumption tests and the ratio of adrenal gland/body weight,and its thickness were examined by HE stain. Results Compared with control group, rats subjected to CUMS exhibited increased ratio of adrenal gland /body weight ( P < 0. 01), less sucrose consumption( P<0.01) and inhibited in the open-field test( P<0.01) as well as more despair time in the forced swimming test( P<0.01). While compared with stressed group,treatments with proanthocyanidins (25,50,100 mg·kg-1, po ,21 days) could significantly improve the activities in open-field test ((39.6±3.4) vs (49±4.5), (52.6±3.7),(54.1±1.8) ;all P<0.01) and sucrose consumption( (5.8±2.5)ml vs (8.1±3.3)ml,(8.5±4.1) ml, (9.2±2.6) ml; P < 0.05, P < 0.05, P < 0.01 respectively); Meanwhile, it could reduce the duration time in forced swimming test significantly( (103.5±10.2)s vs (83.7±8.8)s,(75.8±5.9)s,(67.2±6.5)s; all P<0.01) as well as thickness of the adrenal gland(P<0.05, P<0.01).Conclusions This study suggests that the proanthocyanidins (25,50,100 mg·kg-1) has an antidepressant-like effects in CUMS rats. The antidepressant actions of proanthocyanidins, in some degrees, may be related with the regulation of the adrenal gland's structure.

Objective To explore the diagnostic value of MRI special use of breast and uhrasonography in axillary lymph node metastasis of early breast cancer.Methods Clinical data of 136 Ⅰ-Ⅲ A breast cancer patients accepted MRI examination before surgery had been retrospectively studied,analysing diagnostic value of MRI and ultrasonography in axillary lymph node metastasis of early breast cancer.Results The sensitivity,specificity,and accuracy obtained by MRI were 83.3％,88.6％ and 86.3％.And these data of ultrasonography were 73.1％,76.7％ and 75.0％.The sensitivity,specificity,and accuracy obtained MRI were better than that of ultrasonography.The sensitivity,specificity,and accuracy of ≥ 50 years old patients were 70.0％,77.8％ and 75.0％.And ＜ 50 years old patients were 85.7％,92.3％ and 88.9％.The sensitivity,specificity,and accuracy of ＜ 50 years old patients were better than ≥ 50 years old patients.Conclusions The MRI special use of breast have an important value in axillary lymph node metastasis of early breast cancer,especially to gounger than 50 years old patients.It can provide a scientific basis of the clinical accurate treatment for early breast cancer patients.

Objective To evaluate the feasibility and accuracy of three-dimensional (3D) printed left atrial appendage (LAA) models based on 3D transesophageal echocardiography (3D-TEE) data and the application value for treating special anatomic LAA.Methods Data of 18 patients of atrial fibrillation who were underwent LAA occlusion were retrospectively analyzed,including 3D-TEE and CT volume data of the patients.The 3D-TEE data of the LAA were post-processed and a flexible material was used to print the LAA model by 3D printer.The morphological classification and lobulated classifications of LAAs were assessed by the 3D printed models.The measurements of long axis,short axis and depth of LAAs were also performed.And the measurement and classification results were compared with those based on 3D-TEE and CT volume images.A occluder release test was performed on the 3D printed models for patients with challenging LAA morphology.Results For all 18 patients,3D-TEE full volume data of the LAA were successfully reprocessed and printed as 3D LAA models.The consistency of morphological classifications and lobulated classifications of LAAs based on 3D printed models and cardiac CT were 0.92 and 0.83,respectively.No significant differences of LAA ostium dimensions (long axis and short axis) and depth were found between the measurement results based on 3D printed models and 3D-TEE (all P＞0.05).A simulation of LAA occlusion rehearsal was successfully performed on 3D models of two challenging cases.Conclusion The echocardiographic 3D printing technique has high feasibility and accuracy,and can be promising for personalized planning in cases of transcatheter special morphological LAA occlusion.

Objective To improve the canines myocardial infarction curative effect by using ultrasound targeted microbubble destruction (UTMD) combining with nuclear localization signal(NLS) peptide to increase hAng-1 gene transfection efficiency.Methods Forty-six canines were randomly divided into 4 groups (n=10 in each group) after the models of myocardial infarction were prepared.Group A were untreated control;Group B were transfected with hAng-1;Group C were transfected with UTMD+hAng-1;Group D were transfected with UTMD+NLS nuclear localization signal+hAng-1.The therapeutic agents were intravenously injected at one week after myocardial infarction in each group,and the ultrasound were irradiated at the precardium in group C and D.①Echocardiography was used before and at one week after myocardial infarction and 28 days after gene transfection.Two-dimensional echocardiography was used to detect cardiac function,the left ventricular ejection fraction,the left ventricular wall motion and the myocardial contrast echocardiography were used to detect myocardial perfusion of all canines in the four groups.②On twenty-eight days after gene transfection,mRNA and Western Blot were used to detect the expression of hAng-1.Immunohistochemistry was used to detect capillary density of peri-infarct area and microvessel density (MVD).Masson′s trichromatic staining and gross specimen were used to evaluate the degree and the area of myocardial fibrosis.The gene transfection efficiency and the curative effect of all the four groups were evaluated and compared.Results ①Before myocardial infarction,in four groups canine ventricular wall motion and cardiac function were normal,and myocardial filling defect was not showed by myocardial contrast echocardiography.At one week after myocardial infarction,the left ventricular anterior and interval walls motion and the left ventricular ejection fraction in the four groups were significantly decreased.Myocardial contrast echocardiography showed anterior and interval walls myocardial filling defect.There was no significant difference among the four groups(P>0.05);On 28 days after gene transfection the left ventricular ejection fraction in the four group were increased in an order of group A,B,C,D,there was significant difference when comparing group C and D with other groups separately(P<0.05).Myocardial contrast echocardiography showed much contrast filling in the infarction and surrounding area in group D,a little contrast filling in group C and filling defect in group A and group B,there was significant difference when comparing group C and D with other groups separately(P<0.05).②RT-PCR and Western Blot showed the hAng-1 mRNA and protein expression in group D were higher than those in the other group.There was significant difference when comparing group C and group D with other groups separately(P<0.05).③Immunofluorescence showed the capillary densities were(4.7±1.6)/mm2,(11.2±2.8)/mm2,(70.0±6.4)/mm2 and (85.3±7.0)/mm2 in group A,group B,group C and group D.The differences were statistically significant compared group C and group D with other groups (P<0.05).④Masson′s trichromatic staining and cardiac gross specimen showed that the degree and area of myocardial fibrosis were gradually reduced in an order of group A,B,C,D.Conclusions UTMD and NLS peptide could effectively transfect hAng-1 gene and it provided a novel strategy of gene treatment for ischemic heart disease.

Objective:To meet the individualized needs of residents in standardized residency training of cardiac intervention specialty for cardiac ultrasound teaching, to construct a bionic 3D printing model of ultrasound source, and to discuss its application value in ultrasound teaching and simulation exercises.Methods:Sixty residents majored in cardiovascular medicine receiving standardized residency training of ultrasound in ultrasound imaging department of Renmin Hospital of Wuhan University were randomized into experimental group and control group in average. The experimental group adopted ultrasound bionic 3D printing heart model combined with problem-based learning (PBL) teaching method, and the control group adopted traditional multimedia PPT combined with PBL teaching method. At the end of teaching activities, theoretical examination, operation assessment and questionnaire survey were conducted to evaluate and compare the teaching effect between the two groups. SPSS 21.0 software was performed for t test. Results:The theoretical examination scores of the experimental group and the control group were (81.4±8.2) points and (74.8±9.4) points, respectively, P=0.002, and the operation assessment scores were (89.1±5.6) points and (71.5±8.8) points, respectively, P<0.001. The questionnaire survey showed that the experimental group had better feedbacks than the control group in the aspects of mastery of heart anatomy knowledge, learning experience, learning interest, learning efficiency, understanding of interventional heart disease, clinical skills of interventional operation and overall satisfaction (all P<0.05). Conclusions:Ultrasound bionic 3D printing combined with PBL teaching method can help strengthen the mastery of cardiac anatomy knowledge, promote the learning experience of cardiac ultrasound and improve the skills of interventional operation. The teaching effect is significant and the students' satisfaction is high.