Objective:To study and establish the quality standard of Polyoxyl 15 hydroxystearate(HS15),a phar-maceutical excipient,and systematically evaluate its functionality-related characteristics and safety.HS15 was applied to the preparation of docetaxel(DTX)injection to further investigate the safety and pharmacokinetic char-acteristics of the injection in vitro and in vivo.Methods:Based on the general USP-NF2024,EP11.0 and the fourth general rules of the Chinese Pharmacopoeia 2020 edition,the quality standards of HS15 were studied.Combined with the functional properties of surfactants,the critical micelle concentration of HS15 was investigated,and its safety was investigated by hemolysis test and vascular irritation test in vitro.HS15 was further applied to the preparation of DTX injection,and the safety and efficacy of the preparation were comprehensively evaluated by in vitro cytotoxicity test and in vivo pharmacokinetic study.Results:According to the experimental results and the pharmacopoeia of various countries,the quality standard of HS15 was preliminarily formulated.When the concen-tration of HS15 was 1 mg·mL-1,the hemolysis rate was about0.2%,the vascular irritation was small,and the DTX injection was safe in vitro and in vivo.The pharmacokinetic behavior was in line with expectations.Conclusion:This study successfully established the quality standard of HS15,and its functional correlation index research and safety evaluation strategy can provide reference for the quality control of similar excipients.The appli-cation of HS15 in the preparation of DTX injection provides a theoretical and experimental basis for its application in the development of insoluble antitumor drug injection.

Objective:To study and establish the quality standard of Polyoxyl 15 hydroxystearate(HS15),a phar-maceutical excipient,and systematically evaluate its functionality-related characteristics and safety.HS15 was applied to the preparation of docetaxel(DTX)injection to further investigate the safety and pharmacokinetic char-acteristics of the injection in vitro and in vivo.Methods:Based on the general USP-NF2024,EP11.0 and the fourth general rules of the Chinese Pharmacopoeia 2020 edition,the quality standards of HS15 were studied.Combined with the functional properties of surfactants,the critical micelle concentration of HS15 was investigated,and its safety was investigated by hemolysis test and vascular irritation test in vitro.HS15 was further applied to the preparation of DTX injection,and the safety and efficacy of the preparation were comprehensively evaluated by in vitro cytotoxicity test and in vivo pharmacokinetic study.Results:According to the experimental results and the pharmacopoeia of various countries,the quality standard of HS15 was preliminarily formulated.When the concen-tration of HS15 was 1 mg·mL-1,the hemolysis rate was about0.2%,the vascular irritation was small,and the DTX injection was safe in vitro and in vivo.The pharmacokinetic behavior was in line with expectations.Conclusion:This study successfully established the quality standard of HS15,and its functional correlation index research and safety evaluation strategy can provide reference for the quality control of similar excipients.The appli-cation of HS15 in the preparation of DTX injection provides a theoretical and experimental basis for its application in the development of insoluble antitumor drug injection.

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

Objective:To investigate the clinicopathological features, diagnosis and prognosis of diffuse leptomeningeal glioneuronal tumor (DLGNT).Methods:Five cases of DLGNT diagnosed from January 2016 to January 2020 were collected from Xuanwu Hospital, Capital Medical University. The clinical features, histopathologic characteristics, immunohistochemical and molecular genetic findings and prognosis were analyzed and the relevant literature was reviewed.Results:The five patients (two males and three females) were aged 2 to 52 years (median 11 years), and had history of increased intracranial pressure (headache and vomiting) or limb weakness. Three of them were younger than 16 years of age. The imaging studies showed diffuse intracranial and intraspinal nodular leptomeningeal thickening and enhancement, with or without parenchymal involvement. At times there were associated small cyst-like lesions. Imaging interpretations were inflammatory lesions in three cases and space occupying lesions in two. Microscopically, in three cases the tumors showed low to moderate cellularity, consisting of relatively monomorphous oligodendrocyte-like cells arranged in small nests or diffusely distribution. No mitosis and necrosis were observed. In two cases there were increased cellularity with a diffuse honeycomb pattern. The tumor showed mild to moderate polymorphism with hyperchromatic nuclei. Mitosis, endothelial vascular proliferation and glomeruloid vessels were seen. Necrosis was absent. The tumor cells in all five cases were positive for synaptophysin,Olig2 and negative for IDH1 and H3 K27M. GFAP was focally positive in four cases and only one case expressed NeuN partly. The Ki-67 labeling index was 1%-35%. BRAF fusion was detected in four cases. Genetic analysis showed solitary 1p deletion in two cases (2/5), while all cases were negative for 1p/19q co-deletion (0/5). The five patients were followed up for 13 to 28 months (median 15 month). One patient died after 27 months. There was no evidence of tumor progression in the remaining four patients.Conclusions:DLGNT is rare and easily confused with other central nervous system tumors and inflammatory lesions. Therefore, the diagnosis of DLGNT should be made based on comprehensive information including imaging, morphologic and corresponding immunohistochemical examinations and molecular genetics to avoid misdiagnosis and delay in management.

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

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 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

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

Objective To investigate the clinical application value of combined detection of ALK fusion gene and c?ros oncogene 1 receptor tyrosine kinase ( ROS1) fusion gene in non?small cell lung cancer ( NSCLC) using real?time fluorescent PCR. Methods A kit for combined detection of ALK fusion gene and ROS1 fusion gene based on fluorescent PCR was used to simultaneously detect the two fusion genes in 302 cases of NSCLC specimens. The results were validated through Sanger sequencing. The consistency of the two detection methods was analyzed. Results All 302 cases of NSCLC specimens were successfully analyzed through fluorescent PCR (302/302). 12 cases (4.0%) were found to contain ALK fusion gene, including 3 cases with ALK?M1, 3 with ALK?M2, 3 with ALK?M3, 1 with ALK?M4, and 2 with ALK?M6 fusion gene. 12 cases (4.0%) were found to contain ROS1 fusion gene, including 1 case with ROS1?M7, 8 cases with ROS1?M8,1 case with ROS1?M12,1 case with ROS1?M14,and 1 case with double?positive ROS1?M3 and ROS1?M8 fusion genes. The total detection rate of ALK fusion gene and ROS1 fusion gene was 7. 9%(24/302) and 278 cases showed to be negative for ALK fusion gene and ROS1 fusion gene. The successful detection rates for Sanger DNA sequencing were also 100%. The positive, negative and total coincidence rates obtained by real?time fluorescent PCR and by Sanger DNA sequencing were all 100%. Conclusions The results of Sanger DNA sequencing demonstrate that the real?time fluorescent PCR assay is equally effective in detecting ALK and ROS1 fusion genes in NSCLC tissues. Furthermore, real?time fluorescent PCR assay can be used to detect trace ALK and ROS1 fusion gene simultaneously in tiny samples, and can save time and avoid repeated sampling. It is worthy of recommendation as a rapid and reliable detection technique.

Objective To investigate the clinical application value of combined detection of ALK fusion gene and c?ros oncogene 1 receptor tyrosine kinase ( ROS1) fusion gene in non?small cell lung cancer ( NSCLC) using real?time fluorescent PCR. Methods A kit for combined detection of ALK fusion gene and ROS1 fusion gene based on fluorescent PCR was used to simultaneously detect the two fusion genes in 302 cases of NSCLC specimens. The results were validated through Sanger sequencing. The consistency of the two detection methods was analyzed. Results All 302 cases of NSCLC specimens were successfully analyzed through fluorescent PCR (302/302). 12 cases (4.0%) were found to contain ALK fusion gene, including 3 cases with ALK?M1, 3 with ALK?M2, 3 with ALK?M3, 1 with ALK?M4, and 2 with ALK?M6 fusion gene. 12 cases (4.0%) were found to contain ROS1 fusion gene, including 1 case with ROS1?M7, 8 cases with ROS1?M8,1 case with ROS1?M12,1 case with ROS1?M14,and 1 case with double?positive ROS1?M3 and ROS1?M8 fusion genes. The total detection rate of ALK fusion gene and ROS1 fusion gene was 7. 9%(24/302) and 278 cases showed to be negative for ALK fusion gene and ROS1 fusion gene. The successful detection rates for Sanger DNA sequencing were also 100%. The positive, negative and total coincidence rates obtained by real?time fluorescent PCR and by Sanger DNA sequencing were all 100%. Conclusions The results of Sanger DNA sequencing demonstrate that the real?time fluorescent PCR assay is equally effective in detecting ALK and ROS1 fusion genes in NSCLC tissues. Furthermore, real?time fluorescent PCR assay can be used to detect trace ALK and ROS1 fusion gene simultaneously in tiny samples, and can save time and avoid repeated sampling. It is worthy of recommendation as a rapid and reliable detection technique.