Pathological features and clinicopathological significance of TERT promoter mutation in breast fibroepithelial tumors without definite diagnosis / 中华病理学杂志

Objective: To investigate the pathological features and the clinicopathological significance of TERT detection in those tumors that were difficult to diagnosis. Methods: A total of 93 cases of fibroepithelial tumors without definite diagnosis were collected from the Affiliated Hospital of Qigndao University between 2013 and 2021. The clinical details such as patients' age and tumor size were collected. All slides were re-reviewed and the pathologic parameters, including stromal cellularity, stromal cell atypia, stromal cell mitoses, and stromal overgrowth were re-interpreted. Sanger sequencing was used to detect TERT promoter status, and immunohistochemistry was performed to detect TERT protein expression. The relationship between TERT promoter mutation as well as protein expression levels and the clinicopathological parameters were also analyzed. Results: The patients' ages ranged from 30 to 71 years (mean of 46 years); the tumor size ranged from 1.2 to 8.0 cm (mean 3.8 cm). These tumors showed the following morphologic features: leafy structures in the background of fibroadenoma, or moderately to severely abundant stromal cells. The interpretations of tumor border status were ambiguous in some cases. The incidence of TERT promoter mutation was high in patients of age≥50 years, tumor size≥4 cm, and stromal overgrowth at ×4 or ×10 objective, and these clinicopathologic features were in favor of diagnosis of phyllodes tumors. TERT protein expression levels was not associated with the above clinicopathologic parameters and its promoter mutation status. Conclusions: The diagnostic difficulty for the breast fibroepithelial tumors is due to the difficulty in recognition of the leafy structures or in those cases with abundant stromal cells. A comprehensive evaluation combined with morphologic characteristics and molecular parameters such as TERT promoter may be helpful for the correct diagnosis and better evaluating recurrence risk.

Effects of HMGB1 on Proliferation and Secretion of Human Bone Marrow Mesenchymal Stem Cells / 中国实验血液学杂志

OBJECTIVE@#To investigate the effect of high mobility group protein 1(HMGB1) on the proliferation and cytokine expression of human bone marrow mesenchymal stem cells (MSC).@*METHODS@#Different concentrations of recombinant human HMGB1 protein (100, 200, 400, 800 and 1000 ng/ml) were incubated with MSC for 24, 48, 72 h and the proliferation of MSC were detected respectively by using the CCK-8 method and flow cytometry. The best concentrations of HMGB1 incubated with MSC was determined (200 ng/ml, 1000 ng/ml), and the flow cytomerty was used to determine the effect of HMGB1 on the proliferation of MSC. The mRNA expression levels of IL-10, TGF- β1, TSG-6 and IFN-γ in MSC incubated with HMGB1 protein were detected by real-time quantitative PCR and ELISA.@*RESULTS@#The result of MSC identification and flow cytometry showed that the CD105, CD73 and CD90 were expressed, but did not expression CD45, CD34, CD11b, CD19 and HLA-DR; CCK-8 showed that HMGB1 at the concentrations of 100 ng/ml, 200 ng/ml and 400 ng/ml could promote the proliferation of MSC incubated for 24, 48 and 72 h as compared with the control group (P<0.05), and the most effective concentration was 200 ng/ml; flow cytometry showed that the compared with the control group, HMGB1 200 ng/ml could induce MSC from G1 phase to S phase to promote the proliferation of MSC; QPCR showed that the mRNA expression of MSC cytokines IL-10, TGF-β1, TSG-6 increased while IFN-γ decreased at the concentration of 200 ng/ml HMGB1 as compared with the control group. ELISA experiments showed that the HMGB1 200 ng/ml acting on MSC for 48 h could significantly promoted the secretion of IL-10, TGF-β 1 and TSG-6(P<0．05), while IFN-γ showed no significant difference as compared with control group.@*CONCLUSION@#Recombinant human HMGB1 can promote the proliferation and secretion of MSC in healthy people.

Effect of hypnosis on pain and fear in the acupuncture subjects / 中国针灸

OBJECTIVE@#To observe the effect of hypnosis on pain and fear in the healthy acupuncture subjects.@*METHODS@#A total of 52 healthy subjects were randomized into an observation group and a control group, 26 cases in each one. In the observation group, the subjects received the first-time acupuncture under hypnosis. After wakened up and 30 min later, the subjects received the second-time acupuncture under clear consciousness condition. In the control group, the subjects received the first-time acupuncture under clear consciousness condition, 30 min later, received the second-time acupuncture under hypnosis. Likert scale was adopted to investigate the relaxation, pain sensation and the willingness in the subjects at the normal condition before acupuncture, after the first-time and the second-time acupuncture of the two groups separately. Using Boeran electronic blood pressure monitor, the pulse and blood pressure were measured in the subjects at the normal condition and after the first-time acupuncture of the two groups.@*RESULTS@#Compared with the normal condition, the relaxation degree was increased, the pain sensation decreased and the willingness enhanced after acupuncture either after hypnosis or after wakened-up in the observation group (0.05). In the control group, compared with the normal condition, after the first-time acupuncture (acupuncture in clear consciousness), the relaxation degree was decreased, pain was alleviated and the willingness was increased when acupuncture was exerted once again (0.05); compared with the first-time acupuncture, the relaxation degree was increased, pain alleviated and willingness enhanced after the second-time acupuncture (acupuncture after hypnosis) (0.05). In the control group, compared with the normal condition, the pulse was faster, both the diastolic pressure and systolic pressure were increased after the first-time acupuncture (<0.05, <0.01). In the observation group, compared with the normal condition, the pulse was getting slow and blood pressure was reduced after the first-time (acupuncture under hypnosis, <0.01). Compared with the first-time acupuncture in the control group, pulse was getting slow and blood pressure was reduced in the observation group after acupuncture under hypnosis (<0.001).@*CONCLUSION@#During acupuncture, with hypnosis combined, the fear alleviates, pain reduces and the willingness of acupuncture increases in the subjects.

Expression of HMGB1 in Spleen of Adult Patients with Chronic and Refractory Immune Thrombocytopenia and Its Significance / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the expression and clinical significance of high mobility group box 1(HMGB1) in spleen of adult patients with chronic and refractory immune thrombocytopenia(ITP).</p><p><b>METHODS</b>Twenty chronic and refactory ITP patients received splenectomy were enrolled in ITP group and 20 cases of traumatic spleen rupture were enrolled in control group. The splenectomy efficacy in ITP patients was analyzed retrospectively. The HMGB1 expression in spleen tissue was detected by immunohistochemistry, and the correlation between different expression levels of HMGB1 and splenectomy efficacy were analysed. Meanwhile, the protein expression levels of HMGB1 in peripheral blood serum and mononuclear cells(PBMNC) of 25 patients with chronic and refractory ITP were detected by ELISA and Western blot.</p><p><b>RESULTS</b>The median platelet count before splenectomy was 7.5 (0-20) ×10/L; all the patients showed that the initial response to splenectomy within the first month after operation was 100%, the median time of response was 1 day (1-6 days). The median peak platelet count post splenectomy was 448.5 (161-1272)×10/L. In the median time of 10(3-30) months, the platelets count in 8 patients was reduced to varying degrees. After a median follow-up of 69.5 months (22-195), complete response was found in 12 patients, 4 cases showed response and 4 did not. The HMGB1 expression positive rate in spleen of ITP patients was significantly higher than that in control group (85.0% vs 15.0%)(P<0.001). There were a negative correlation between the HMGB1 expression in ITP and therapeutic outcome after splenectomy (r=-0.791, P<0.01). In addition, HMGB1 expression levels in serum and PBMNC of the patients with chronic refractory ITP were also significantly higher than that in healthy controls (P<0.01).</p><p><b>CONCLUSION</b>The splenectomy has been found to be effective therapeutic method for patients with ITP, the HMGB1 highly express in the spleen of the patients with chronic refractory ITP, but negatively correlats with the therapeutic outcome after splenectomy.</p>

Investigation of Enterobius vermicularis infection in children in Jiangxi Province in 2014 / 中国血吸虫病防治杂志

To know the status of Enterobius vermicularis infection in children in Jiangxi Province in 2014, so as to provide the evidence for the formulation of prevention and control measures.A survey was performed according to the scheme of the 3rd Principal Human Parasites of Jiangxi Province in 2014. Based on the ecological regions, a stratified cluster sampling method was applied by the economic and geographic situation. There were 84 survey sites from 28 counties, and the basic data were also collected in the different investigation sites, and the round-end tube adhesive cellophane anal swab was used to examine E. vermicularis eggs for the children aged 3–6 years.A total of 1 486 children aged 3-6 years were detected, the E. vermicularis infection rate was 13.73% (204/1 486), and the infection rates were 13.89% (114/821) and 13.53% (90/665) in the male and female, respectively. The infection rate in the different age groups showed a gradual rise then fall trend, the lowest infection rate was 10.05% (38/378) in the 3-year age group, and the highest infection rate was 18.24% (81/444) in the 5-year age group. The infection rates in the high, medium and low-income survey sites were 13.79% (87/631), 17.23% (51/296), and 11.81% (66/559), respectively. The E. vermicularis infection rates in the 4 ecological regions were from 12.34% to 17.74%, but there was no significant difference among the different ecological regions (P > 0.05).The status of E. vermicularis infection in children in Jiangxi Province is relatively serious, and therefore, the parasitic disease control sectors should continue to strengthen the monitoring and control work of E. vermicularis infection in children.

Direct modulation of firing activity by dopamine Dlike receptors in the globus pallidus of both normal and parkinsonian rats / 生理学报

The globus pallidus occupies a critical position in the indirect pathway of the basal ganglia circuit, which regulates movement under both normal and pathological conditions. Previous studies have shown that the globus pallidus receives dopaminergic innervation from the axonal collaterals of nigrostriatal fibers. Both dopamine Dand Dlike receptors are expressed in the globus pallidus. The present study was aimed to investigate the direct in vivo electrophysiological effects of dopamine Dlike receptors in the globus pallidus of both normal and parkinsonian rats. Extracellular recordings of multi-barreled microelectrode were used in the present study. In normal rats, micro-pressure ejection of dopamine Dlike receptor agonist quinpirole induced different effects on the firing rate of globus pallidus neurons. In 24 out of the 61 pallidal neurons, quinpirole significantly increased the firing rate by (62.7 ± 11.2)%. In another 16 neurons, quinpirole decreased the spontaneous firing rate by (37.5 ± 2.9)%. Furthermore, co-application of dopamine Dlike receptor antagonist, sulpride, blocked quinpirole-induced modulation of the firing rate of pallidal neurons. On the 6-hydroxydopamine (6-OHDA) lesioned side of parkinsonian rats, quinpirole increased the firing rate in 25 out of the 47 pallidal neurons by (64.2 ± 10.1)%, while decreased the firing rate in 11 neurons by (51.9 ± 6.2)%. Our findings suggest that activation of pallidal dopamine Dlike receptors may bidirectionally modulate the spontaneous firing of globus pallidus neurons in both normal and parkinsonian rats.

Role of Treg Cells in Pathogensis of Mouse ITP / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the role of Treg cells in the pathogenesis of idiopathic thrombocytopenic purpura (ITP).</p><p><b>METHODS</b>The ITP mouse model was established, the Treg cell ratio in peripheral blood and spleen was detected by flow cytometry, the CD4+ CD25+ T cells were sorted by immunomagnetic beads, the Treg cell associated transcription factors (Foxp3, Smad7, STAT5 and Akt-1) and cytokines (IL-10, TGF-β) in CD4+ CD25+ T cells were enriched from spleen mononuclear cells, and the mRNA expression of Treg cell was measured by real-time PCR.</p><p><b>RESULTS</b>The ratio of Tregs in peripheral blood and spleen decreased significantly in ITP mouse, as compared with the controls (P<0.01). In addition, the mRNA expression of IL-10, TGF-β and Foxp3 decreased significantly in spleen CD4+ CD25+ T cells (P<0.05). Expression of Smad 7 mRNA was higher than that of controls.</p><p><b>CONCLUSION</b>The alteration in Treg frequency and function may be responsible for the immune dysfunction in ITP disease. It is also speculated that the lower mRNA expression of Foxp3 and higher mRNA expression of Smad 7 may inhibit the proliferation and differentiation of Treg cells.</p>

Left ventricular systolic strain of the cardiac allograft evaluated with three-dimensional speckle tracking echocardiography / 华中科技大学学报（医学）（英德文版）

Three-dimensional speckle tracking echocardiography was employed to evaluate the changes of left ventricular systolic strain in 23 heart transplant recipients at 1st, 3rd, 6th and 12th month after heart transplantation, and 23 healthy subjects served as controls. The three-dimensional full-volume echocardiographic images of left ventricle were recorded and then were analyzed using EchoPAC software. The strain curves and peak systolic strain values for each segment and overall left ventricular wall were obtained. Left ventricular global peak longitudinal strain (GPSL), global peak radial strain (GPSR), global peak circumferential strain (GPSC) and global peak area strain (GPSA) were measured and then statistically analyzed. There were no significant differences in left ventricular ejection fraction (LVEF) and cardiac output (CO) between heart transplant recipients and controls. The GPSL in heart transplant recipients at 1st month after surgery was significantly lower than that in controls, but close to the normal value at 3rd month after surgery and later. The GPSC, GPSA and GPSR were significantly lower in heart transplant recipients at 1st, 3rd, 6th and 12th month after surgery than those in controls. It is suggested that three-dimensional speckle tracking echocardiography can be used for monitoring changes of left ventricular systolic strains and evaluating left ventricular systolic function in cardiac allograft.

Assessment of left ventricular function in patients with aortic regurgitation using tow-dimensional speckle-tracking echocardiography / 中华医学超声杂志（电子版）

Objective Analyse the change of left ventricular (LV) longitudinal and radial strain in patients with aortic regurgitation (AR) and discuss the relationship between the 2D strain parameter and the filling and ejection of LV. Methods Thirty healthy controls and 45 patients with AR (24 patients with moderate AR and 21 with severe AR) were enrolled in this study, LV systolic global peak radial strain(GRS), systolic global peak longitudinal strain(GLS) and systolic peak longitudinal strain(S), systolic peak longitudinal strain rate(SRs), early diastolic peak longitudinal strain rate(SRe) of every segment were measured or calculated using 2D-STE, early and late diastolic transmitral flow velocity (E, A) were recorded by pulsed Doppler echocardiography and early diastolic mitral annular velocity (Ea) were assessed by tissue Doppler imaging,the E/A and E/Ea ratio were calculated. Discuss the relationship of GLS and LV ejection fraction (LVEF), GLS and E/Ea using the Pearson correlation analysis. Results The GLS were (-20.09±1.47)%, (-18.68±1.52)%, (-12.56±3.25)%and the GRS were (46.71±7.65)%, (43.01±5.95)%, (28.52±6.13)% in control group, patients with moderate and severe AR (MAR group and SAR group) respectively. There were significant differences among the groups (F =82.08,47.69, both P < 0.01) as following:SAR group with control group and MAR group [ q=17.56,13.60 (GLS), q=13.44, 10.20 (GRS), all P<0.01),MAR group and control group [ q=3.42 (GLS), P<0.01]. The SRs of the apical segment were (-1.24±0.22)s-1, (-1.19±0.25)s-1, (-1.04±0.28)s-1 in control group,MAR group and SAR group respectively. There were significant differences among the groups (F=4.47, P < 0.05) as following:SAR group with control group and MAR group ( q=4.02,3.28, both P<0.01). The S, SRe of apical segment and the S,SRs,SRe of basal and midventricular in MAR group were all lower than the control group ( q=4.42, 5.01, 3.48, 3.24, 4.78, 4.12, 3.61, 6.72, all P < 0.01). Pearson correlation analysis revealed the GLS had a relationship with LVEF and E/Ea ( r=-0.73, 0.64, both P<0.01). Conclusion The reduced longitudinal strain and strain rate could detect LV dysfunction in patients with AR in early stage and the GLS had the ability to reflect the diastolic filling and systolic ejecting of the LV.

Left ventricular systolic strain of the cardiac allograft evaluated with three-dimensional speckle tracking echocardiography / 华中科技大学学报（医学）（英德文版）

Three-dimensional speckle tracking echocardiography was employed to evaluate the changes of left ventricular systolic strain in 23 heart transplant recipients at 1st, 3rd, 6th and 12th month after heart transplantation, and 23 healthy subjects served as controls. The three-dimensional full-volume echocardiographic images of left ventricle were recorded and then were analyzed using EchoPAC software. The strain curves and peak systolic strain values for each segment and overall left ventricular wall were obtained. Left ventricular global peak longitudinal strain (GPSL), global peak radial strain (GPSR), global peak circumferential strain (GPSC) and global peak area strain (GPSA) were measured and then statistically analyzed. There were no significant differences in left ventricular ejection fraction (LVEF) and cardiac output (CO) between heart transplant recipients and controls. The GPSL in heart transplant recipients at 1st month after surgery was significantly lower than that in controls, but close to the normal value at 3rd month after surgery and later. The GPSC, GPSA and GPSR were significantly lower in heart transplant recipients at 1st, 3rd, 6th and 12th month after surgery than those in controls. It is suggested that three-dimensional speckle tracking echocardiography can be used for monitoring changes of left ventricular systolic strains and evaluating left ventricular systolic function in cardiac allograft.

Analysis of CD4(+)CD25(+)CD127(low/-) Treg cells in mice / 中国实验血液学杂志

The quantitative identification and enrichment of viable regulatory T cells (Treg) requires reliable surface markers that are selectively expressed on Treg. Foxp3 is the accepted marker of natural Treg, but it cannot be used to isolate cells for functional studies. CD127 is a new surface marker expressed in Treg cells. In this study, two populations of Treg, including CD4(+)CD25(+)CD127(low/-) and CD4(+)CD25(+)Foxp3(+)T cells, and profiles of the Foxp3 expression in CD4(+)CD25(+)CD127(low/-) cells were compared to evaluate which population is better. The peripheral blood cells were collected and spleen suspension of BALB/C mice were prepared, and using triple staining CD4, CD25, CD127 and CD4, CD25, Foxp3. The profiles of Treg, including CD4(+)CD25(+)CD127(low/-) and CD4(+)CD25(+)Foxp3(+) were detected by flow cytometry. The quadruple staining CD4, CD25, Foxp3 and CD127 were used to determine the CD127 expression in CD4(+)CD25(+)Foxp3(+) cells. The results showed that on T cell subset the median expression levels of CD4(+), CD4(+)CD25(+) were 39.02%, 5.35% in peripheral blood and 23.49%, 3.86% in spleen. On CD4(+) T cell subset, the median expression level of CD4(+)CD25(+)CD127(low/-) and CD4(+)CD25(+)Foxp3(+)T cells were 7.13%, 3.97% in peripheral blood and 12.8%, 8.23% in spleen. The ratio of CD4(+)CD25(+)CD127(low/-) T cells was higher than that of CD4(+)CD25(+)Foxp3(+) cells in both peripheral blood and spleen cells (P < 0.01). The CD4(+)CD25(+)CD127(low/-) cells highly expressed Foxp3, while the CD4(+)CD25(+)Foxp3(+)T cells lowly expressed CD127. It is concluded that compared with the CD4(+)CD25(+)Foxp3(+) populations, CD4(+)CD25(+)CD127(low/-) T cells better fit the definition of naturally occurring regulatory T cells in peripheral blood cells and spleen of BALB/C mice. CD127(low/-) is a characteristic marker on surface of CD4(+)CD25(+) Treg cells, and has been confirmed to be more specific marker for quantitatively sorting Treg cells.

Inhibition of osteosarcoma cell proliferation by a short hairpin RNA targeting proliferation cell nuclear antigen / 中华病理学杂志

<p><b>OBJECTIVE</b>To construct a plasmid vector expressing the short hairpin RNA (shRNA) targeting proliferation cell nuclear antigen (PCNA), and to investigate its effect in vitro on the expression of PCNA, proliferation and apoptosis of human osteosarcoma cells.</p><p><b>METHODS</b>A plasmid vector expressing the short hairpin RNA targeting at PCNA was constructed and transfected into human osteosarcoma cell line MG-63 by dosper liposomal method. PCNA mRNA and protein expressions were examined using RT-PCR and immunohistochemical staining, respectively. Inhibition of the cell proliferation was studied by MTT method and colony forming assay. DNA synthesis was analyzed by (3)H-TdR incorporation and the cell cycle was determined by flow cytometry. The apoptotic cells were stained with acridine orange.</p><p><b>RESULTS</b>Expression of PCNA mRNA after the transfection was markedly inhibited by 80.51% with a PI value of 25.68% of that of the control group. PCNA shRNA inhibited MG-63 cell growth detected by using MTT method, with an inhibition rate of 61.78% at 48 h. DNA synthesis rate also decreased in the (3)H-TdR incorporation test. Flow cytometry analysis showed an increase of the percentage of G(0)/G(1) phase cells, along with a decrease of cell population in the S phase. The apoptosis rate of cells transfected with the plasmid vector was 16.54%.</p><p><b>CONCLUSIONS</b>PCNA shRNA significantly suppresses the expression of PCNA at both mRNA and protein levels, corresponding to an inhibition of the proliferation of MG-63 cell and an increase of the cellular apoptosis.</p>