Objective To study the changes and their value of ultrasonic myocardial integrated backscatter (IBS)in normal control and patients with super-acute phase of myocardial infarction. Methods There were 38 cases of acute myocardial infarction(AMI)patients in the super-acute phase(time ＜ 2 h infarction)and the acute phase(infarction time ＞ 2 h,typical ECG changes)for myocardial ultrasonic backscatter parameters detection,an additional 25 cases as healthy control group. The myocardial infarction region and non-infracted myocardial tissue region average duration of cardiac cycle of integrated backscatter(IBS)were measured,and IBS adjusted value (IBS%)was calculated as the ratio of myocardial IBS to the pericardium,the difference of IBS at end-diastolic and late systolic was used as cyclic variation of IBS(CVIB). The ratio of IBS to pericardial CVIB was used as its adjusted value(CVIB%). Over the same period for the electrocardiogram,myocardial enzymes and cardiac troponin I were measured. Results When ECG was not typically changed in patients with super-acute phase AMI,the IBS values significantly increased in the myocardial infarction region than the normal control group (43. 7 ± 10. 8)dB vs.(22. 6 ± 4. 6)dB,P ＜0.01),and CVIB was significantly lower than that in the normal control group(10. 2 ±2. 6)dB vs.(13. 2 ± 3.8)dB,P ＜ 0.01]. The IBS in the acute phase in patients was significantly higher than that in the normal control group and those non-infarcted areas,and CVIB was significantly lower than that in the normal control group and those in the non-infarcted areas. The changes were consistent with the ECG changes. Conclusions Ultrasonic backscatter parameters might be helpful for diagnosis of hyperacute period of AMI,and the determination of the scope and extent of AMI.

BACKGROUND: The inflammatory reaction occurs following implantation of cardiovascular stent with manifestations of the activation of blood coagulation system and dramatically increasing of inflammatory markers serum C-reactive protein. OBJECTIVE: To explore the changes of inflammatory reaction and C-reactive protein in coronary artery disease patients following cardiovascular stent implantation.METHODS: A computer-based online search of CNKI (1990/2009) and PubMed databases (1990/2009) was performed for related articles with the key words "cardiovascular stent, C-reactive protein" in Chinese and "cardiovascular stent on plasma, C-reactive protein" in English.RESULTS AND CONCLUSION: Based on the metal stents, drug-eluting stents can transfer the active drugs to the damaged vessels, release them into the vascular wall and inhibit the in-stent restenosis, Main drugs of anti-inflammatory drug-eluting stent include dexamethasone and methylprednisolone. Main drugs of anti-migratory and anti-proliferative drug-eluting stent includerapamycin, paclitaxel and actinomycin D. Main drugs of supporting intima concrescence stent include estradiol. Coronary artery stents implantation can induce and aggravate local inflammation reaction, which have important infection for vascular endodermis hyperplasia and restenosis occurrence. Some impressible index for inflammation reaction, such as levels of C-reactive protein,have predictive value for vascular events following the coronary artery stents implantation. A significant increase in plasma C-reactive protein after coronary stenting has been observed following stent implantation. Therefore, understanding of inflammatory reaction and C-reactive protein, as well as cytokine changes is important for preventing restenosis, early treatment of restenosis, as well as improving treatment effect.

We evaluated the patient in the early,advanced or healing phase of the disease by MRI in the treatment of chronic brucellosis spondylitis and to guide the clinical treatment.MRI images of 40 patients with clinically diagnosed chronic brucellosis spondylitis were analyzed retrospectively.The imaging findings of early,advanced and healing patients were summarized.MRI showed abnormal signals in the vertebral body,intervertebral disc,paraspinaI and psoas muscle.It is early stage if the intervertebral space was normal,and advanced stage if combined with interverbral space stenosis.It demonstrated short T1 and short T2 signal or similar to the normal vertebral body,combined with intervertebral space stenosis,for the healing stage.According to the specific imaging manifestations of chronic brucellar spondylitis in the course of disease development,it is possible to evaluate the clinical stage of the disease and guide the rational selection of clinical treatment.

Objective To compare the diagnostic values of US-guided optical imaging system (OPTIMUS) and full-field digital mammography (FFDM) in breast neoplasms. Methods Sixty-four patients suspected of breast neoplasms underwent OPTIMUS and FFDM before surgery. All patients were proved histopathologically. The accuracy of diagnosis was compared with chi-square test. Results Sixty-five masses were delineated by OPTIMUS and FFDM. The diagnostic accuracy, sensitivity and specificity of OPTIMUS were 86. 2% ( 56/65 ), 87.0% ( 20/23 ) and 85.7% ( 36/42 ), respectively. The positive likelihood ratio was 6. 09 and the negative likelihood ratio was 0. 175. The diagnostic accuracy, sensitivity and specificity of FFDM were 73.8% (48/65), 82. 6% ( 19/23 ) and 71.4% (30/42), respectively. The positive likelihood ratio was 2. 892 and the negative likelihood ratio was 0. 243. There were no significant differences between OPTIMUS and FFDM ( x2 = 3. 077 ,P ＞ 0. 05). Conclusion OPTIMUS is similar and supplementary to FFDM for the diagnosis of breast neoplasms.

Objective:To observe the effects of berberine on procoagulant and fibrinolytic inhibitory factors produced by rat type Ⅱ alveolar epithelial cell (AECⅡ) induced by lipopolysaccharide (LPS).Methods:AECⅡ cells (RLE-6TN cells) were cultured in vitro, and the cells in logarithmic growth phase were collected. The cytotoxicity text of berberine was detected by cell counting kit-8 (CCK-8) to determine the drug concentration range according to inhibition concentration of half cells (IC 50). The RLE-6TN cells were divided into five groups, the cells in blank control group were cultured in DMEM; the cells in LPS group were stimulated with 5 mg/L LPS; and the cells in berberine pretreatment groups were pretreated with 20, 50 and 80 μmol/L berberine for 1 hour, and then were co-cultured with 5 mg/L LPS. The cells were collected after LPS induced for 24 hours. The protein and mRNA expression levels of tissue factor (TF), tissue factor pathway inhibitor (TFPI) and plasminogen activator inhibitor-1 (PAI-1) in the cells were detected by Western blotting and real-time fluorescence quantification reverse transcription-polymerase chain reaction (RT-qPCR). The levels of activated protein C (APC), precollagen Ⅲ peptide (PⅢP), thrombin-antithrombin complex (TAT) and antithrombin Ⅲ (ATⅢ) in the cell supernatant were measured by enzyme linked immunosorbent assay (ELISA). Results:According to the inhibition rate curve, the IC 50 of berberine on RLE-6TN cells was 81.16 μmol/L. Therefore, 20, 50 and 80 μmol/L were selected as the intervention concentration of berberine. Compared with the blank control group, the expression and secretion of procoagulant and fibrinolytic inhibitory factors were abnormal in RLE-6TN cells after LPS induced for 24 hours. The protein and mRNA expression levels of TF and PAI-1 in the LPS group were significantly increased, but the protein and mRNA expression levels of TFPI were significantly decreased. Meanwhile, the levels of APC and ATⅢ in the cell supernatant were significantly decreased, while the levels of PⅢP and TAT were significantly increased. After pretreatment with berberine, the abnormal expression and secretion of procoagulant and fibrinolytic inhibitory factors induced by LPS were corrected in a dose-dependent manner, especially in 80 μmol/L. Compared with the LPS group, the protein and mRNA expression levels of TF and PAI-1 in the berberine 80 μmol/L group were significantly decreased [TF protein (TF/GAPDH): 0.45±0.02 vs. 0.55±0.03, TF mRNA (2 -ΔΔCt): 0.39±0.08 vs. 1.48±0.11, PAI-1 protein (PAI-1/GAPDH): 0.37±0.02 vs. 0.64±0.04, PAI-1 mRNA (2 -ΔΔCt): 1.14±0.29 vs. 4.18±0.44, all P < 0.01] and those of TFPI were significantly increased [TFPI protein (TFPI/GAPDH): 0.53±0.02 vs. 0.45±0.02, TFPI mRNA (2 -ΔΔCt): 0.94±0.08 vs. 0.40±0.05, both P < 0.01]. Meanwhile, the levels of APC and ATⅢ in the cell supernatant were significantly increased [APC (μg/L): 1 358.5±26.0 vs. 994.2±23.1, ATⅢ (μg/L): 118.0±7.4 vs. 84.4±2.7, both P < 0.01], while those of PⅢP and TAT were significantly decreased [PⅢP (μg/L): 11.2±0.4 vs. 18.6±0.9, TAT (ng/L): 222.1±2.8 vs. 287.6±7.0, both P < 0.01]. Conclusions:Berberine could inhibit the LPS-induced expressions of procoagulant and fibrinolytic inhibitory factors in rat AECⅡ cells and promote the expressions of anticoagulant factors in a dose-dependent manner. Berberine may be a new therapeutic target for alveolar hypercoagulability and fibrinolysis inhibition in acute respiratory distress syndrome (ARDS).

Objective:To determine the effect of andrographolide (AD) on the expression of procoagulant and fibrinolytic inhibitory factors in rat type Ⅱ alveolar epithelial cells (AECⅡ) stimulated by lipopolysaccharide (LPS).Methods:The AECⅡ cells RLE-6TN in the logarithmic growth phase were divided into 5 groups: the normal control (NC) group, the LPS group, and the 6.25, 12.5, and 25 mg/L AD groups (AD 6.25 group, AD 12.5 group, AD 25 group). The NC group was cultured with RPMI 1640 conventional medium. In the LPS group, 5 mg/L LPS was added to the RPMI 1640 conventional medium for stimulation. Cells in the AD groups were treated with 6.25, 12.5, and 25 mg/L AD in advance for 1 hour and then given LPS to stimulate the culture. The cells and cell culture supernatant were collected 24 hours after LPS stimulation. The protein and mRNA expressions of tissue factor (TF), tissue factor pathway inhibitor (TFPI), and plasminogen activator inhibition-1 (PAI-1) in cells were detected by Western blotting and real-time fluorescent quantitative polymerase chain reaction (RT-qPCR). The levels of procollagen Ⅲ peptide (PⅢP), thrombin-antithrombin complex (TAT), antithrombin Ⅲ (AT-Ⅲ) and activated protein C (APC) in the cell supernatant were detected by enzyme linked immunosorbent assay (ELISA).Results:Compared with the NC group, the protein and mRNA expressions of TF and PAI-1 in the LPS group were significantly increased, and the protein and mRNA expressions of TFPI were significantly reduced. At the same time, the levels of PⅢP and TAT in the cell supernatant were significantly increased, the levels of AT-Ⅲ, APC were significantly reduced. Compared with the LPS group, the protein and mRNA expressions of TF and PAI-1 in AD 6.25 group, AD 12.5 group, AD 25 group were significantly reduced [TF/GAPDH: 0.86±0.08, 0.45±0.04, 0.44±0.04 vs. 1.32±0.10, TF mRNA (2 -ΔΔCt): 2.59±0.25, 2.27±0.05, 1.95±0.04 vs. 4.60±0.26, PAI-1/GAPDH: 2.11±0.07, 1.45±0.04, 0.86±0.09 vs. 2.56±0.09, PAI-1 mRNA (2 -ΔΔCt): 3.50±0.22, 2.23±0.29, 1.84±0.09 vs. 6.60±0.27, all P < 0.05], while the protein and mRNA expressions of TFPI were significantly increased [TFPI/GAPDH: 0.78±0.05, 0.81±0.03, 0.84±0.07 vs. 0.36±0.02, TFPI mRNA (2 -ΔΔCt): 0.46±0.09, 0.69±0.07, 0.91±0.08 vs. 0.44±0.06, all P < 0.05]. Also the levels of PⅢP and TAT in the cell supernatant were significantly reduced, and the levels of AT-Ⅲ and APC were significantly increased [PⅢP (μg/L): 13.59±0.23, 12.66±0.23, 10.59±0.30 vs. 15.82±0.29, TAT (ng/L): 211.57±6.41, 205.69±4.04, 200.56±9.85 vs. 288.67±9.84, AT-Ⅲ (μg/L): 102.95±3.86, 123.92±2.63, 128.67±1.67 vs. 92.93±3.36, APC (μg/L): 1 188.95±14.99, 1 366.12±39.93, 1 451.15±29.69 vs. 1 145.55±21.07, all P < 0.05]. With the increase of the dose of AD, the above-mentioned promotion and inhibition effects became more obvious. In the AD 25 group, TF, PAI-1 protein and mRNA expressions decreased, TFPI mRNA expression increased, PⅢP level in the supernatant decreased and AT-Ⅲ, APC levels increased compared with AD 6.25 group, the difference was statistically significant, and the decrease of PAI-1 protein expression and PⅢP level in the supernatant were also statistically significant compared with AD 12.5 group. Conclusions:Andrographolide in the dose range of 6.25-25 mg/L can dose-dependently inhibit the expression and secretion of procoagulant and fibrinolytic inhibitor-related factors in AECⅡ cells RLE-6TN stimulated by LPS, and promote the secretion of anticoagulant factors. 25 mg/L has the most obvious effect.

Pulldown assay is an in vitro method for studies of protein-protein interactions, in which tagged proteins are usually expressed as the bait to enrich other proteins that could bind to them. In this technology, the GST tag is broadest used for its modest size and hydrophilic property. In most cases, the GST tag could increase the hydrophility of the fusion protein and help to avoid the formation of inclusion bodies. However, in the other few cases, the target protein may be strongly hydrophobic or have complicated structures that were hard to fold and assemble in correct conformations without champerons, and even the existence of GST tag could not make them soluble. These proteins were always expressed as inclusion bodies and had no functions. LMO2 was a small molecular weight and insoluble protein, in this study, GST system and MBP system were used to express GST-LMO2 and MBP-LMO2 fusion proteins, respectively. We found that GST-LMO2 fusion protein was expressed as inclusion bodies whereas MBP-LMO2 fusion protein was expressed in soluble form. Moreover, the production rate of MBP-LMO2 was also much higher than GST-LMO2. Then MBP-LMO2 fusion proteins and renatured GST-LMO2 fusion proteins were used as bait in pulldown assay to study the interaction between LMO2 and endogenous GATA1 in K562 cells. Western blot analyses showed that both of these proteins could bind to endogenous GATA1 in K562 cells, but recovered GATA1 protein by MBP-LMO2 fusion protein was much more than GST-LMO2 fusion protein. These results suggest that using of MBP system is a helpful attempt in the case of studying small molecular weight, strong hydrophobic proteins.
Adaptor Proteins, Signal Transducing ; Carrier Proteins ; chemistry ; Chemical Precipitation ; DNA-Binding Proteins ; chemistry ; GATA1 Transcription Factor ; chemistry ; Genetic Vectors ; Glutathione Transferase ; chemistry ; Humans ; K562 Cells ; LIM Domain Proteins ; Maltose-Binding Proteins ; Metalloproteins ; chemistry ; Protein Binding ; Protein Interaction Domains and Motifs ; Protein Renaturation ; Proto-Oncogene Proteins ; chemistry ; Recombinant Fusion Proteins ; genetics ; metabolism

Bone morphogenetic protein 2 (BMP2), which belongs to the transforming growth factor-beta (TGF-beta) superfamily, is a multifunctional molecule with distinct abilities to induce bone formation. BMP2 has been identified to have eminent pharmaceutical importance for clinical application. We previously constructed stable cell line in Chinese hamster ovary cells (CHO) that highly expressed recombinant human BMP2 (rhBMP2). For large-scale production of the recombinant protein used in clinical application, it is critical to have both high expression and stability of the protein. In the present study, the stability of the cell line (rCHO(hBMP2)-C8) with the highest expression, as well as the stability of rhBMP2 protein were investigated systematically. We cultured the rCHO (hBMP2)-C8 cell line in the presence or absence of MTX for two months, the cell growth and rhBMP2 production characteristics were examined during the culture; we found the duration that the rCHO(hBMP2)-C8 cell line could secret rhBMP2 continually into the serum-free medium. Moreover, we detected the temperature sensitivity of rhBMP2 in culture medium. This study will contribute to our understanding for further producing rhBMP2 by large-scale culture technology.
Animals ; Bone Morphogenetic Protein 2 ; biosynthesis ; genetics ; CHO Cells ; Cell Culture Techniques ; methods ; Cricetinae ; Cricetulus ; Culture Media ; Genetic Vectors ; genetics ; Humans ; Recombinant Proteins ; biosynthesis ; genetics

Objective:To explore the mediating effect of self-control between smartphone addiction and academic procrastination in medical undergraduates.Methods:A total of 640 medical college undergraduates were selected by convenient sampling method. The self-designed general information questionnaire, smartphone addiction proneness scale (SAPS), brief self-control scale (BSCS) and procrastination assessment scale-students (PASS) were conducted among the students. SPSS 25.0 was used for descriptive statistical analysis, independent sample t test, one-way ANOVA and Pearson correlation analysis. Results:Correlation analysis showed that PASS scores were positively correlated with SAPS scores ( r=0.29, P<0.01), and negatively corrected with BSCS scores ( r=-0.26, P<0.01); the SAPS scores were negatively corrected with BSCS scores ( r=-0.33, P<0.01). Mediating effect analysis showed that the mediating role of self-control between smartphone addiction and academic procrastination were significant (effect size=0.13, 95%CI=0.03-0.26), and the mediating effect accounted for 38.24%. Conclusion:Self-control played partial mediating effect between smartphone addiction and academic procrastination in medical undergraduates. In order to improve the current situation of medical undergraduate's academic procrastination, medical college educators can intervene from the perspective of smartphone addiction and self-control.

Objective:To explore the efficacy and safety of low- and medium-dose of glucocorticoids in adult patients with acute respiratory distress syndrome (ARDS) through Meta-analysis and trials sequential analysis (TSA).Methods:Databases associated with adult ARDS treatment with low- and medium-dose glucocorticoids both in English and in Chinese were searched from PubMed, Medline, China Biology Medicine (CBM), Cochrane Library, CNKI, Wanfang Data and VIP, of which the search duration was from the establishment of the database to December 2020. Low-dose glucocorticoids were defined as methylprednisolone ≤ 1 mg·kg -1·d -1, and medium dose glucocorticoids were defined as methylprednisolone ≤ 2 mg·kg -1·d -1. According to the Cochrane Collaboration bias risk assessment tool, the quality of the included literature was evaluated, and the data were extracted. Meta-analysis and TSA were used to evaluate the effects of low- and medium-dose glucocorticoids on the hospital mortality, intensive care unit (ICU) mortality, and mechanical ventilation free time in ICU for 28 days, PaO 2/FiO 2, and the occurrence of nosocomial infections and hyperglycemia. Results:A total of 996 patients in 7 literatures were finally included, including 515 patients in the low- and medium-dose glucocorticoid group (hormone group) and 481 patients in the conventional treatment group (control group). The research quality of 7 literatures was relatively high. The results of Meta-analysis and TSA showed that, compared with the control group, the hospital mortality in the hormone group was significantly decreased [relative risk ( RR) = 0.77, 95% confidence interval (95% CI) was 0.66-0.89, P = 0.000 6], and mechanical ventilation free time in ICU for 28 days was significantly prolonged [standardized mean difference ( SMD) = 0.50, 95% CI was 0.36-0.65, P < 0.000 1]. Although Meta-analysis showed that the ICU mortality of the hormone group was significantly lower than that of the control group ( RR = 0.61, 95% CI was 0.38-0.99, P = 0.04), the TSA results showed that the cumulative Z value crossed the traditional threshold, but did not cross the TSA cut-off value, and the sample size did not reach required information size (RIS, n = 3 252), needed more research to confirm. Although Meta-analysis showed that PaO 2/FiO 2 in the hormone group was significantly higher than that in the control group ( SMD = 0.78, 95% CI was 0.13-1.43, P = 0.02), TSA showed that the cumulative Z value did not pass the traditional and TSA cut-off values. More research was needed for verification. Meta-analysis also showed that there was no significant difference in the incidence of new infection ( RR = 0.93, 95% CI was 0.74-1.17, P = 0.54) and the incidence of hyperglycemia ( RR = 1.11, 95% CI was 1.00-1.23, P = 0.05) between the hormone group and the control group. Conclusion:low- and medium-dose of glucocorticoids therapy can reduce the hospital mortality of adult ARDS patients and shorten the mechanical ventilation duration in ICU for 28 days, and low- and medium-dose of glucocorticoids therapy does not increase the risk of infection and hyperglycemia.