Objective To study the clinical efficacy of cell block yielded by ultrasound-guided fine needle aspiration in the early diagnosis of thyroid cancer and differential diagnosis of thyroid follicular tumor.Methods Cytological smears were performed in 864 thyroid nodules by ultrasound-guided fine needle aspiration.Then the residual specimens were made as cell blocks.Diagnosis was made by observation of the organization structure of the cell block.Cell block sections from 78 nodules with diagnosis of follicular tumors were applied to the detection of immunocytochemical staining for minichromosome maintenance protein (MCM2), growth arrest and DNA damage-induced nuclear protein 153 (GADD153), and Ki-67.Results Satisfaction rate of cytological smear combined with cell block specimen was 94.3 ％ (815/864) and accuracy rate of the diagnosis was 94.0％ (766/815), which were higher than in cytological smear group (P＜0.01).The false negative rate of cytological smear combined with cell block specimen was 16.6％ (41/247)in diagnosis of thyroid cancer.Positive rate of M CM2 and GADD153 in thyroid follicular neoplasia were lower than that of follicular carcinoma (P ＜ 0.01).There was no significant difference between thyroid follicular neoplasia and follicular carcinoma in regard to positive rate of Ki-67 (P ＞ 0.05).Conclusions The combination of ultrasound-guided fine needle aspiration-yielded cell block and molecular markers has great significance in early diagnosis of thyroid cancer and differentially diagnosis of thyroid follicular tumor.

OBJECTIVEChronic airway inflammation is associated with the polarization of TH2 cells in asthma. Prostaglandin D2 (PGD2) plays an important role in the polarization of TH2 cells. This study aimed to investigate the changes of PGD2 receptors (DP1/CRTH2) on T lymphocytes and their significance in asthma.

METHODSSeventy-two children with asthma were assigned to two groups: acute attack (n=42) and remission (n=30). Thirty-five healthy children were used as the control group. Plasma levels of TH2 cytokines IL-4 and IL-5, and TH1 cytokine INF-gamma were detected using ELISA. Radiological binding assay (RBA) was used to measure the contents of DP1/CRTH2 receptors on T cells in peripheral blood (PPB).

RESULTSThe total combining contents of DP and CRTH2 on T cells in PPB in the acute attack and the remission groups were significantly higher than those in the control group (p<0.01). There was no significant difference in the DP1 content among the three groups. Serum levels of IL-4 and IL-5 significantly increased (p<0.01), in contrast, serum levels of TH1 cytokine IFN-gamma were significantly reduced in the acute attack and the remission groups compared with those in the control group (p<0.01).

CONCLUSIONSThe total combining contents of DP and CRTH2 on T cells increased, serum levels of TH2 cytokines also increased, but serum levels of TH1 cytokine decreased significantly in the acute attack and the remission phases in children with asthma. This showed that a polarization of TH2 cells occurred in children with asthma and suggested that CRTH2 antagonism may be a new target for the treatment of asthma.

Asthma ; etiology ; immunology ; therapy ; Child ; Child, Preschool ; Chromosome Mapping ; Cytokines ; blood ; Female ; Humans ; Male ; Receptors, Immunologic ; blood ; Receptors, Prostaglandin ; blood ; T-Lymphocytes ; chemistry ; Th2 Cells ; immunology

OBJECTIVE: To explore the effects of siRNA hsa-circ-0000885 modified bone marrow mesenchymal stem cells (BMSCs) on osteogenic differentiation, cell proliferation and apoptosis in order to provide new ideas and methods for the clinical treatment of osteoporosis (OP). METHODS: From September 2018 to February 2020, 13 patients with osteoporosis admitted to our hospital were selected as the research objects, including 11 females and 2 males, with an age of (65.45±10.77) years old. After obtaining the informed consent of patients, peripheral blood tissues were extracted. Then the expression level of cir-cRNA in peripheral blood mononuclear cells(PBMC) was detected by circ RNA chip. The expression of circ RNA was silenced by siRNA technology. The BMSCs were transfected with lentivirus. According to the siRNA interference plasmid hsa-circ-0000885, the cells were divided into the blank group, the empty vector group and the siRNA interference group. After 72 hours of treatment, the cell cycle was detected by flow cytometry, the apoptosis level was detected by AV-PI kit, and the osteogenic differentiation ability of BMSCs was detected by ALP staining. RESULTS: The expression of hsa-circ-0000885 in PBMC of patients with osteoporosis was significantly higher than that of healthy controls ( CONCLUSION The lentivirus mediated siRNA hsa-circ-0000885 plasmid transfected into BMSCs and osteoclast co culture system can promote cell proliferation, inhibit apoptosis and promote osteogenic differentiation of BMSCs, which can be used as a potential therapeutic target for OP patients.
Aged ; Apoptosis/genetics* ; Cell Differentiation ; Cell Proliferation/genetics* ; Cells, Cultured ; Coculture Techniques ; Female ; Humans ; Lentivirus ; Leukocytes, Mononuclear ; Mesenchymal Stem Cells ; Middle Aged ; Osteoclasts ; Osteogenesis/genetics* ; RNA, Small Interfering/genetics* ; Transfection

OBJECTIVE: To investigate the effect of RUNX2 gene overexpression vector modified exosomes derived from bone marrow mesenchymal stem cells (BMSCs) combined with calcium carbonate scaffold system in bone defect. METHODS: Rabbit BMSCs were used as the research object, and BMSCs were identified by flow cytometry. Construct RUNX2 gene overexpression vector, transfect BMSCs with lentivirus, and collect exosomes by ultracentrifugation. The morphology of exosomes was observed by transmission electron microscope, the expression of exosome marker CD63 was detected by Western blot, and the calcium carbonate scaffold was constructed by three chamber parallel automatic temperature control reaction system. According to whether the RUNX2 gene overexpression vector was transfected or not, the complex of BMSCs and calcium carbonate scaffold was divided into three groups, namely BMSCs group, RUNX2 overexpression group and exosome group. The osteogenic differentiation of BMSCs was detected by oil red O staining and RT-PCR. There were 9 clean adult healthy male New Zealand white rabbits, aged (12.97±1.21) months, with a body weight of (19.3±3.6) kg, with 3 rabbits in each group. The animal model of skull defect was constructed by surgical method, and the repair of bone defect was evaluated by imaging, he staining and Masson staining. RESULTS: The results of flow cytometry showed that the expression of CD29 protein, CD44 protein, CD11b protein and CD45 protein on the surface of BMSCs were 99.5%, 100%, 0.1% and 0.1%, respectively. Transmission electron microscopy showed that the exosomes were bilayer vesicles with a diameter of 50 to 150 nm. Western blot showed that the molecular marker CD63 of exosomes was positive. Oil red O staining showed that the osteogenic differentiation of BMSCs in exosome group was significantly higher than that in RUNX2 overexpression group and BMSCs group. The results of RT-PCR showed that the relative expressions of RUNX2, BMP-2 and ALP mRNA in BMSCs in exosome group were significantly higher than those in RUNX2 overexpression group and BMSCs group (P<0.05). The imaging results showed that the repair effect of skull defect in exosome group was better than that in RUNX2 overexpression group. HE staining and Masson staining showed that the repair effect of skull defect in exosome group was better than that in RUNX2 overexpression group (P<0.05). MSCs in exosome group was significantly higher than that in RUNX2 overexpression group and BMSCs group. The results of RT-PCR showed that the relative expressions of RUNX2, BMP-2 and ALP mRNA in BMSCs in exosome group were significantly higher than those in RUNX2 overexpression group and BMSCs group(P<0.05). The imaging results showed that the repair effect of skull defect in exosome group was better than that in RUNX2 overexpression group. HE staining and Masson staining showed that the repair effect of skull defect in exosome group was better than that in RUNX2 overexpression group(P<0.05). CONCLUSION Compared with RUNX2 gene overexpression vector transfection, extraction of exosomes directly can promote the differentiation of BMSCs into osteoblasts more efficiently, and the combination with calcium carbonate scaffold can better promote the healing of bone defects. So as to provide new ideas and methods for the clinical treatment of bone defects.
Animals ; Calcium Carbonate/metabolism* ; Core Binding Factor Alpha 1 Subunit/metabolism* ; Exosomes/metabolism* ; Humans ; Male ; Osteogenesis/genetics* ; RNA, Messenger/metabolism* ; Rabbits

OBJECTIVETo explore whether the chemotactic factor CCL5 is the major factor of diffuse large B cell lymphoma (DLBCL) induced by diabetes or not.

METHODSThe normal human B cells and DLBCL cells were cultured in vitro; the RT-PCR was used to detect their CCL5 mRNA expression, the human DLBCL cell line and mouse-derived DLBCL cell line A20 were cultured in vitro by using glucose at 5 and 30 mmol/L, respectively, then their CCL5 mRNA expression was detected by PT-PCR; the diabetic mouse model was constructed through peritoneal injection of streptozotocin at low dose in the BALB/c mice; the cell lines with stably high and low expression of CCL5 were established via lentiviral transfection and the cell lines with low and high expression of CCL5 were subcutaneously injected into diabetic mice and mice with normal blood sugar level. According to blood sugar level, the experimental mice were divided into 2 groups: diabetic group (A group) and normal blood sugar group as control (B group); then according to CCL5 expression level, the A group and B group were divided as well into high expression group (A1 group and B1 group) and low expression group (A2 group and B2 group), respectively, the tumor-formation rate, tumor-formation time, tumor size and texture were analyzed, respectively; the CCL5 expression was detected by using HE staining of tumor tissue and immunohistochemical method.

RESULTSThe expression of CCL5 mRNA in human DLBCL cell line cultured in vitro was higher than that in normal B cells (P < 0.05); the expressions of CCL5 mRNA in human DLBCL cells cultured in high sugar concentration in vitro and mouse DLBCL cells were higher than those in cells cultured in low sugar concentration (P < 0.05). The tumor-formation rates in diabetic mice injected with high and low expression of CCL5 cell line A20 were 93.3% in A1 group and 60% in A2 group; the their tumor-formation time was 7.0 ± 0.85 days in A1 group and 9.5 ± 2.8 days in A2 group, while the tumor formation rates in mice with normal blood sugar level were 20% in B1 group and 20% in B2 group, and their tumor-formation time was 12 ± 1.3 days and 14 ± 2.5 days respectively; the CCL5 expression level in tumor tissue of diabetic mice was higher than that in tumor tissue of mice with normal blood sugar level.

CONCLUSIONThe high blood glucose level can casase increase of DLBCL risk and promote the tumor growth; the chemotactic factor CCL5 may play an important role in the growth and migration of DLBCL caused by diabetes mellitus.

Animals ; B-Lymphocytes ; metabolism ; Cell Line, Tumor ; Chemokine CCL5 ; metabolism ; Diabetes Mellitus, Experimental ; Disease Models, Animal ; Humans ; Lymphoma, Large B-Cell, Diffuse ; metabolism ; Mice ; Mice, Inbred BALB C ; RNA, Messenger

OBJECTIVETo investigate the clinical significance of cell blocks obtained by ultrasound-guided fine needle aspiration in diagnosing parotid gland masses.

METHODSCell blocks were made in 285 parotid gland masses by ultrasound-guided fine needle aspiration. Diagnosis was conducted using the cell blocks. Non-tumor masses were subjected to conservative treatment, and cysts and tumors were treated with surgery. The cell block sections from masses with the diagnosis of adenoid cystic carcinoma (ACC) and pleomorphic adenoma (PA) were applied to the detection of immunocytochemical staining for the stem cell factor receptor CD117.

RESULTSThe satisfaction rate of the specimen was 95.1% (271/285). The accuracy rate of the diagnosis was 94.5% (256/271), the sensitivity was 87.0% (67/77), and the specificity was 98.1% (157/160). The positive rate of CD117 in ACC was 95.2% (20/21), whereas that in PA was 20.3% (25/123). The positive rate of CD117 in ACC was higher than that in PA (P<0.01).

CONCLUSIONSThe use of cell blocks obtained from ultrasound-guided fine needle aspiration, together with molecular marker detection, has great significance in diagnosing parotid gland masses.
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Adenoma, Pleomorphic ; Biopsy, Fine-Needle ; Carcinoma, Adenoid Cystic ; Cysts ; Female ; Humans ; Male ; Parotid Gland ; Parotid Neoplasms ; Personal Satisfaction ; Staining and Labeling ; Ultrasonics

Background and Objectives: Mesenchymal stromal cells (MSCs)-based treatment for degeneration of intervertebral disc (IVD) has been proposed recently. We here addressed whether MSC secreted factors can rejuvenate nucleus pulposus- derived stem/progenitor cells from degenerated disc (D-NPSCs) in vitro. Methods: and Results: We analyzed the expression of MSCs and NP cell specific surface markers, pluripotency related genes, multilineage potential and cell proliferative capacity of D-NPSCs upon incubation with the conditioned medium which was collected from the umbilical cord derived MSCs (UCMSCs). Our results indicated that the conditioned medium restore the stemness of D-NPSCs by up-regulating the expression level of CD29 and CD105, pluripotency related genes OCT4 and Nanog, and NP progenitor marker Tie2. The increased stemness was accompanied by promoted cell proliferative capacity and improved osteogenic and chondrogenic differentiation potential. Conclusions Our findings suggested that the UCMSCs derived conditioned medium might be used to rejuvenate the degenerated NP stem/progenitor cells.

OBJECTIVETo evaluate the validity and reliability of Diabetes Self-management Knowledge, Attitude, and Behavior Assessment Scale (DSKAB).

METHODSWe selected 460 patients with diabetes in the community, used the scale which was after two rounds of the Delphi method and pilot study. Investigators surveyed the patients by the way of face to face. by draw lots, we selected 25 community diabetes randomly for repeating investigations after one week. The validity analyses included face validity, content validity, construct validity and discriminant validity. The reliability analyses included Cronbach's α coefficient, θ coefficient, Ω coefficient, split-half reliability and test-retest reliability.

RESULTSThis study distributed a total of 460 questionnaires, reclaimed 442, qualified 432. The score of the scale was 254.59 ± 28.90, the scores of the knowledge, attitude, behavior sub-scales were 82.44 ± 11.24, 63.53 ± 5.77 and 108.61 ± 17.55, respectively. It had excellent face validity and content validity. The correlation coefficient was from 0.71 to 0.91 among three sub-scales and the scale, P<0.001. The common factor cumulative variance contribution rate of the scale and three sub-scales was from 57.28% to 67.19%, which achieved more than 50% of the approved standard, there was 25 common factors, 91 items of the total 98 items held factor loading ≥0.40 in its relevant common factor, it had good construct validity. The scores of high group and low group in three sub-scales were: knowledge (91.12 ± 3.62) and (69.96 ± 11.20), attitude (68.75 ± 4.51) and (58.79 ± 4.87), behavior (129.38 ± 8.53) and (89.65 ± 11.34),mean scores of three sub-scales were apparently different, which compared between high score group and low score group, the t value were - 19.45, -16.24 and -30.29, respectively, P<0.001, and it had good discriminant validity. The Cronbach's α coefficient of the scale and three sub-scales was from 0.79 to 0.93, the θ coefficient was from 0.86 to 0.95, the Ω coefficient was from 0.90 to 0.98, split-half reliability was from 0.89 to 0.95.Test-retest reliability of the scale was 0.51;the three sub-scales was from 0.46 to 0.52, P<0.05.

CONCLUSIONThe validity and reliability of the Diabetes Self-management Knowledge, Attitude, and Behavior Assessment Scale are excellent, which is a suitable instrument to evaluate the self-management for patients with diabetes.

Diabetes Mellitus ; therapy ; Health Knowledge, Attitudes, Practice ; Humans ; Pilot Projects ; Reproducibility of Results ; Self Care ; Surveys and Questionnaires