Objective To investigate the value of ultrasound-guided diffuse optical tomography (US-guided DOT) to predict clinical efficacy of breast cancer neoadjuvant chemotherapy (NAC).Methods Eighty-eight breast cancer patients with 93 lesions were included.Pre-and post-last chemotherapy,the size,total hemoglobin concentration (THC) of each lesion were measured by ultrasonography (US) and US-guided DOT.Based on the guidelines to evaluate the response to treatment in solid tumors,the lesions of treated breast cancer patients were divided into 4 types of responses to NAC:complete response (CR),partial response (PR),stable disease (SD),and progressive disease (PD).Efficient groups include CR and PR groups.Results As expected,no significant difference was found in size and THC for untreated lesions (all P ＞0.05).However,for the treated lesions(P =0.001),THC,pre-vs post-treatment size changes (△Size％) (P =0.002) and THC changes (△THC％) (P ＜0.001) were significantly varied among CR,PR,SD,PD groups.When compared with pre-treated,tumor sizes after treatment were changed significantly in all CR(P ＜0.001),PR(P ＜0.001),SD (P =0.023) and PD (P =0.001),while significant change of THC was only found in CR(P ＜0.001),PR(P ＜0.001) and SD (P =0.002).When △THC％ =23.9％ as the threshold for prediction of NAC efficiency,the area under the curve of ROC was 0.75,and the sensitivity was 73.7％,specificity was 76.5 ％,positive predictive value was 93.3 ％,negative predictive value was 39.4％,accuracy was 74.2％.Conclusions △size％ changes in consistent with △THC％ among the intergroups,but their changes levels are different,the highest change percent appears in CR,gradually decreased in PR,SD groups.△THC％ will contribute to predict preoperative clinical NAC efficacy.

The interaction among collagen, von Willebrand factor (vWF) and glycoprotein Ib axis is the first step in hemostasis and thrombosis, especially under high shear condition. To develop a new remedy of anti-thrombosis, mRNA from endothelial cells was extracted, and reverse transcription PCR was adopted to amplify DNA of interest. After sequencing, recombinant expression vector was constructed. The amplified DNA fragment of vWF domain A3 was inserted into expression vector with 6 x his taq, pET20b(+), the recombinant was transformed into E coli (strain DE3) and induced by IPTG. Recombinant vWF-A3 was designated as a recombinant fragment comprising residues 918 - 1114 of mature vWF subunit. It was purified through Ni-NTA resin column and refolded in Tris buffer containing GSH and GSSG. The results showed that rvWF-A3 was expressed successfully in E coli (strain DE3), accounting for 46% of total bacterial protein with its purity of over 95%. It was identified that rvWF-A3 is capable to bind collagen and inhibit the wild vWF binding to collagen by competition. It is concluded that rvWF-A3 fragment might be an effective antithrombotic agent for preventing arterial thrombosis.
Cloning, Molecular ; Collagen ; metabolism ; Escherichia coli ; genetics ; Humans ; Protein Structure, Tertiary ; Recombinant Proteins ; biosynthesis ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA ; von Willebrand Factor ; chemistry ; genetics ; metabolism

This study was aimed to investigate the expressions of vascular endothelial growth factor (VEGF) and cyclooxygenase-2 (COX-2) in patients with multiple myeloma (MM) and its clinical significance. Expression of VEGF was detected by enzyme linked immunosorbent assay (ELISA) and the level of COX-2 was detected by Western blot. The results showed that the serum VEGF level of multiple myeloma patients (365.34 +/- 65.63 pg/ml) was higher than that in the normal persons (122.52 +/- 39.29 pg/ml) (p < 0.05); the serum VEGF level of patients at advanced stage (395.07 +/- 54.90) pg/ml was higher than those at stable stage (300.33 +/- 44.22) pg/ml (p < 0.05). The serum Cox-2 positive rate in the patients (31%) was higher than that in normal persons (0%) (p < 0.01); the serum Cox-2 positive rate in the patients at advanced stage (50%) was higher than those at stable stage (21%) (p < 0.01). It is concluded that VEGF and COX-2 may play an important role in the pathogenesis and development of multiple myeloma, they can be used to evaluate the status of patients with MM.
Adult ; Aged ; Aged, 80 and over ; Case-Control Studies ; Cyclooxygenase 2 ; blood ; Humans ; Middle Aged ; Multiple Myeloma ; blood ; Vascular Endothelial Growth Factor A ; blood

To study the effect of Cu2+ and Zn2+ on amyloid-beta peptides (Abeta) aggregation, the morphology, size and cell toxicity of Abeta40 aggregates formed with the metal ions have been observed by the methods including ultraviolet spectroscopy, fluorescence spectroscopy and transmission electron microscopy. The results showed that Cu2+ and Zn2+ can accelerate Abeta40 aggregation, and both changed the morphology and size of Abeta40 aggregates. Zn2+ induced Abeta40 to form fibrous Abeta40 aggregates, while the amorphous and fibrous aggregates were produced by the interaction between Cu2+ and Abeta40. In addition, H2O2 was produced when Abeta40 reduced Cu2+. The relationship between metal ions and Abeta40 aggregates was analyzed, and the function of metal ions in Alzheimer's disease (AD) was illustrated in the research.
Amyloid beta-Peptides ; chemistry ; Cell Survival ; drug effects ; Copper ; administration & dosage ; chemistry ; toxicity ; Dose-Response Relationship, Drug ; HeLa Cells ; Humans ; Hydrogen Peroxide ; chemistry ; Ions ; chemistry ; Microscopy, Electron, Transmission ; Peptide Fragments ; chemistry ; Spectrometry, Fluorescence ; Spectrophotometry, Ultraviolet ; Zinc ; chemistry

This study was aimed to further investigate the function of platelet collagen receptor-glycoprotein VI and to screen its specific inhibitor. The extracellular domain of platelet glycoprotein VI (GPVI) in E. coli was expressed by recombinant technology, the extracellular domain cDNA of GPVI was amplified from pBluescript KS(-)-GPVI plasmid by PCR. Proved by sequencing, the expression vector pET-20b(+)-GPVI was constructed, which was then transformed into E. coli (BL21(DE3)pLysS) and induced by IPTG. The recombinant GPVI was purified on Ni-NTA resin column and renatured in PBS containing GSH and GSSG. The anti-penta His McAb and anti-GPVI polyclonal antibody were used to identify the recombinant GPVI in Western blotting. Collagen binding test was conducted to investigate the biological activity of recombinant GPVI. The results showed that the recombinant GPVI was expressed in E. coli and successfully purified, which was confirmed to be similar to the native GPVI in Western blotting. The recombinant GPVI can bind the type I collagen in dose-dependent manner. In conclusion, the recombinant GPVI can be achieved in E. coli and restore its native characteristics after renaturation.
Blood Platelets ; metabolism ; Blotting, Western ; Escherichia coli ; genetics ; Humans ; Integrin alpha2beta1 ; Platelet Membrane Glycoproteins ; biosynthesis ; genetics ; Protein Binding ; Receptors, Collagen ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; isolation & purification

OBJECTIVETo investigate the effect of thalidomide on Annexin II (AnxA2) gene regulation in multiple myeloma cell line RPMI8226 and human microvascular endothelial cell line HMEC-1 cells in vitro, and explore the potential mechanism of thrombosis induced by thalidomide.

METHODSRPMI8226 and HMEC-1 cells were cultivated in vitro. Real time quantitative PCR (RQ-PCR) was used to detect the influence of thalidomide at different concentration on the expression of AnxA2 mRNA, flow cytometry (FCM) and confocal microscopy were used to detect the cell surface protein level after the samples were stimulated with different concentrations of thalidomide.

RESULTSAnxA2 mRNA level in RPMI8226 cells treated with thalidomide at 12.5 microg/ml, 25.0 microg/ml and 50.0 microg/ml was decreased compared with the control group (0.60+/-0.15, 0.33+/-0.14, 0.42+/-0.16, vs 1.07+/-0.16, respectively, P<0.05) and did so in HMEC-1 cells (0.21+/-0.20, 0.08+/-0.08, 0.17+/-0.16 vs 1.16+/-0.24, respectively, P<0.05). The AnxA2 protein level in RPMI8226 cells treated with above mentioned concentrations of thalidomide was also decreased compared with the control (3.39+/-0.32, 2.82+/-0.28, 3.21+/-0.23 vs 5.53+/-0.32, respectively, P<0.05) and that did so in HMEC-1 cells (0.72+/-0.11, 0.64+/-0.08, 0.67+/-0.08 vs 1.40+/-0.15, respectively, P<0.05).

CONCLUSIONSThalidomide can inhibit the expression of AnxA2 mRNA and protein in RPMI8226 and HMEC-1 cells, which may be one of the mechanisms for the development of thrombosis induced by thalidomide in multiple myeloma patients.

Annexin A2 ; genetics ; metabolism ; Cell Line ; Endothelial Cells ; metabolism ; Endothelium, Vascular ; cytology ; Humans ; Multiple Myeloma ; metabolism ; pathology ; RNA, Messenger ; genetics ; Thalidomide ; pharmacology

OBJECTIVETo explore the possible role of CD28/CTLA-4 co-stimulators in immune pathophysiology of acquired aplastic anemia(AA).

METHODSBy FACS, the percentages of CD28, CTLA-4 expressing CD3+ CD4+ T cells and the level of Th1, Th2 in bone marrow were detected in 23 AA patients at active phase, 10 at recovery phase and 15 normal controls. The relationship between the co-stimulators, Th1, Th2, and absolute neutrophil counts (ANC) was evaluated.

RESULTS(1) The percentage of CD28 and CTLA-4 expressing CD3+ CD4+ T cells in bone marrow, and CD28+/CTLA-4+ ratios were (31.40 +/- 10.83)%, (2.45 +/- 1.30)% , and 17.02 +/- 13.44 in normal controls respectively, (39.84 +/- 10.89)%, (1.43 +/- 0.67)%, and 43.04 +/- 37.61 in AA at active phase, respectively, (22 +/- 9.08)%, (3.46 +/- 2.26)%, and 10.49 +/- 7.8 in AA at recovery phase, respectively. The percentage of CD28 and CD28+/CTLA-4+ ratio were significantly higher, while CTLA-4 were lower in active phase AA patients than in normal controls (P < 0.05). These values in recovery phase AA were comparable to those in normal controls. (The Th1, Th2, and Th1/Th2 in bone marrow were (4.21 +/- 2.11)%, (1.99 +/- 1.27)%, and 2.46 +/- 1.28 in normal controls respectively, (11.13 +/- 4. 96)%, (2.46 +/- 1.65)%, and 5.20 +/- 1.98 in active phase AA and (5.39 +/- 4.2)9%, (2.53 +/- 2.41)%, and 2.87 +/- 1.43 in recovery phase AA, respectively. The percentage of Th1 and Th1/Th2 ratio were significantly higher in AA patients at active phase than in normal controls (P < 0.05). (3) The CD28+/CTLA-4+ ratio was positively related to the Th1+ /Th2+ ratio (P < 0.05). ANC was negatively related to CD3+ CD4+ CD28+ T cells (P < 0.01), and positively to CD3 + CD4 ' CTLA-4' T cells (P < 0.01) respectively.

CONCLUSION(1) The expression of CD28 was increased while CTLA-4 decreased on the membranes of CD3+ CD4+ T cells in bone marrow of AA patients. (2) The abnormal expression of CD28 costimulator promoted the shift of immune balance to Thl type. (3) The unbalance of CD28+ / CTLA-4+ is important for the immune pathophysiology of AA.

Adolescent ; Adult ; Aged ; Anemia, Aplastic ; immunology ; metabolism ; Antigens, CD ; immunology ; metabolism ; CD28 Antigens ; immunology ; metabolism ; CD4-Positive T-Lymphocytes ; metabolism ; CTLA-4 Antigen ; Child ; Female ; Humans ; Male ; Middle Aged ; Th1 Cells ; immunology ; Th2 Cells ; immunology

Sodium glucose co-transporter 2 (SGLT-2) inhibitors are newly developed but promising medicine for type 2 diabetes. However, patients with a different renal threshold for glucose excretion (RT(G)) may have a different reaction to this medicine. Therefore, the objective of this study was to investigate the characteristics of RT(G) and its impact factors in patients with type 2 diabetes mellitus (T2DM). The clinical and laboratory data of 36 healthy individuals and 168 in-hospital patients with T2DM were collected and analyzed, RTG was calculated using blood glucose (BG) measured by dynamic BG monitoring, urinary glucose excretion (UGE) and estimated glomerular filtration rate (eGFR). The characteristics of RT(G) were investigated. The risk factors for high RT(G) were analyzed using non-conditional logistic regression analysis. Our results found that RT(G) of the T2DM group was higher than that of the healthy individuals (P < 0.05); and 22.22% from the healthy individuals group but 58.33% from the T2DM group had high RT(G). Age, duration of diabetes, body mass index (BMI), and homeostasis model assessment insulin resistance index (HOMA-IR) were independently associated with high RT(G) (P < 0.05). Further stratified analysis revealed that RT(G) in T2DM patients increased with age, duration of diabetes, and BMI. In conclusion, RT(G) is increased in patients with T2DM, especially in those with longer diabetic duration, higher BMI, and those who are older. Therefore, these patients may be more sensitive to SGLT-2 inhibitors.

Objective: To explore the role of mobile phone addiction and anxiety symptoms in the relationship between childhood psychological abuse and depressive symptoms among college students, in order to provide a basis for mental health promotion. Methods: From February to May 2023, a stratified random sampling method was used to select 1 799 freshmen to juniors from a university in Wuhu City, Anhui Province. The questionnaire survey was conducted using the 2-item Patient Health Questionnaire (PHQ-2), Child Psychological Maltreatment Scale (CPMS), Mobile Phone Addiction Tendency Scale (MPATS), 2-item General Anxiety Disorder (GAD-2). Correlations among each variable were analyzed, and the chain mediating effect of mobile phone addiction and anxiety symptoms was explored. Results: The detection rate of depressive symptoms among college students was 9.7%, and the positive detection rate of childhood psychological abuse was 28.6%. Depressive symptoms were positively correlated with childhood psychological abuse, mobile phone addiction and anxiety symptoms ( r =0.28, 0.32, 0.27, P <0.01). Childhood psychological abuse was positively correlated with mobile phone addiction and anxiety symptoms ( r =0.29, 0.71, P <0.01). Mobile phone addiction and anxiety symptoms were positively correlated ( r =0.30, P <0.01). Childhood psychological abuse could effectively predict depressiove symptoms, mobile phone addiction and anxiety symptoms ( β =0.08, 0.06, 0.66, P <0.01). Mobile phone addiction and anxiety symptoms had a chain mediating effect between childhood psychological abuse and depression symptoms, with a total indirect mediating effect (effect=25.27%, P <0.05), accounting for 72.44% of the total effect. Conclusions Mobile phone addiction and anxiety symptoms play a chain mediating role between childhood psychological abuse and depressive symptoms. Focusing on childhood psychological abuse, mobile phone addiction and anxiety among college students are beneficial for depression symptoms prevention.

OBJECTIVETo investigate the effects of Annexin II (AnxA2) gene silencing by siRNA on proliferation and invasive potential of lymphoma cell line Jurkat cells.

METHODSA synthesized siRNA duplex targeting to AnxA2 was transfected into Jurkat cells. Transfection efficiency was analyzed by real-time PCR and flow cytometry. MTT assay for cell proliferation and transwell plates for invasive potential were performed.

RESULTSCompared with the negative controls, the cell proliferation inhibitory rate of the AnxA2 siRNA transfected Jurkat cells was significantly increased at 24 h, 48 h and 72 h [(17.4 +/- 2.3)%, (22.4 +/- 3.8)%, (37.6 +/- 1.5)% vs (-1.3 +/- 5.1)%, (-5.5 +/- 4.4)%, (-10.8 +/- 5.5)%, respectively, P<0.05]. The cell invasive potential of the transfected Jurkat cells was inhibited remarkably at 48 h (11.3 +/- 4.2 vs 54.3 +/- 8.7, P<0.01).

CONCLUSIONAnxA2 gene silenced by siRNA can inhibit the proliferation and the invasive potential of Jurkat cells remarkably.

Annexin A2 ; genetics ; metabolism ; Cell Proliferation ; Chemotaxis ; genetics ; Gene Silencing ; Humans ; Jurkat Cells ; RNA, Small Interfering ; genetics ; Transfection