Objective:To explore the effects of miR-21-5p exosomes derived from human umbilical cord mesenchymal stem cells on apoptosis of human granular cells.Methods:A granular cell apoptosis model was constructed by treating KGN cells with different concentrations (0 μmol/L, 30 μmol/L, 60 μmol/L, and 100 μmol/L) of phosphoramide nitrogen mustard for 48 h. The mRNA and protein levels of bax and bcl2 were detected using qPCR and Western blotting, respectively. The apoptosis rate was measured using flow cytometry to screen for the optimal concentration of phosphoramide nitrogen mustard for constructing an apoptosis model. Hsa-miR-21-5p overexpression plasmid was used for instantaneously transfecting human umbilical cord mesenchymal stem cells, and the expression level of hsa-miR-21-5p was detected by qPCR. The miR-21-5p exosomes were separated and identified by flow cytometry and electron microscope. Different concentrations (5 μg/mL, 10 μg/mL and 15 μg/mL) of miR-21 exosomes were added into successful KGN cell apoptosis model to overexpress miR-21. The mRNA and protein levels of bax and bcl2 were detected using qPCR and Western blotting, respectively. PKH-26 was used to trace the position of human granular cells. Results:The levels of bax mRNA and protein in KGN cells treated with 60 μmol/L phosphoramide nitrogen mustard were significantly higher than those in the 0 μmol/L phosphoramide nitrogen mustard group (all P<0.001), while the levels of bcl2 mRNA and protein were significantly higher than those in the 0 μmol/L phosphoramide nitrogen mustard group ( P=0.005, P<0.001). The apoptosis rate of KGN cells after 60 μmol/L phosphoramide nitrogen mustard intervention was (38.10±2.90)%, while the apoptosis rate of KGN cells after 30 μmol/L phosphoramide nitrogen mustard intervention was (16.75±2.55)%, they were all significantly higher than that of the 0 μmol/L phosphoramide nitrogen mustard intervention group ( P=0.020, P=0.006). Hsa-miR-21-5p was transiently transfected into human umbilical cord blood mesenchymal stem cells, and the expression of has-miR-21-5p was higher than that in control group detected by qPCR ( P<0.001). The positive rate of surface protein CD9, CD63 and CD81 was 14.9%, 16.4% and 31.4%. The exosome was observed as "tea tray" or "concave hemisphere" by electron microscope. The exosome labeled by PKH-26 entered the granular cells and exerted biological effects. There was no statistically significant difference in bax mRNA expression levels between the 5 μg/mL, 10 μg/mL, and 15 μg/mL empty plasmid exosomes groups and the 60 μmol/L phosphoramide nitrogen mustard group (all P>0.05). However, the expression levels of bax mRNA in the 5 μg/mL, 10 μg/mL, and 15 μg/mL miR-21-5-p exosomes groups were lower than those in the 60 μmol/L phosphoramide nitrogen mustard group ( P=0.008, P=0.003, P<0.001). However, there was no statistically significant difference in the expression of bcl2 mRNA among the groups (all P>0.05). From the perspective of protein levels, there was no statistically significant difference in BAX protein expression between the 5 μg/mL, 10 μg/mL, and 15 μg/mL empty exosomes groups and 60 μmol/L phosphoramide nitrogen mustard group (all P>0.05), while the 5 μg/mL, 10 μg/mL, and 15 μg/mL miR-21-5-p exosomes groups showed a decrease in BAX protein expression compared with the 60 μmol/L phosphoramide nitrogen mustard group, and the differences were statistically significant (all P<0.001). However, there was no statistically significant difference in the expression of BCL2 protein among the intervention groups (all P>0.05). Conclusion:Hsa-miR-21-5p exosomes derived from human umbilical cord blood mesenchymal stem cells can effectively exert the anti-apoptotic effect.

Objective:To explore the effects of miR-21-5p exosomes derived from human umbilical cord mesenchymal stem cells on apoptosis of human granular cells.Methods:A granular cell apoptosis model was constructed by treating KGN cells with different concentrations (0 μmol/L, 30 μmol/L, 60 μmol/L, and 100 μmol/L) of phosphoramide nitrogen mustard for 48 h. The mRNA and protein levels of bax and bcl2 were detected using qPCR and Western blotting, respectively. The apoptosis rate was measured using flow cytometry to screen for the optimal concentration of phosphoramide nitrogen mustard for constructing an apoptosis model. Hsa-miR-21-5p overexpression plasmid was used for instantaneously transfecting human umbilical cord mesenchymal stem cells, and the expression level of hsa-miR-21-5p was detected by qPCR. The miR-21-5p exosomes were separated and identified by flow cytometry and electron microscope. Different concentrations (5 μg/mL, 10 μg/mL and 15 μg/mL) of miR-21 exosomes were added into successful KGN cell apoptosis model to overexpress miR-21. The mRNA and protein levels of bax and bcl2 were detected using qPCR and Western blotting, respectively. PKH-26 was used to trace the position of human granular cells. Results:The levels of bax mRNA and protein in KGN cells treated with 60 μmol/L phosphoramide nitrogen mustard were significantly higher than those in the 0 μmol/L phosphoramide nitrogen mustard group (all P<0.001), while the levels of bcl2 mRNA and protein were significantly higher than those in the 0 μmol/L phosphoramide nitrogen mustard group ( P=0.005, P<0.001). The apoptosis rate of KGN cells after 60 μmol/L phosphoramide nitrogen mustard intervention was (38.10±2.90)%, while the apoptosis rate of KGN cells after 30 μmol/L phosphoramide nitrogen mustard intervention was (16.75±2.55)%, they were all significantly higher than that of the 0 μmol/L phosphoramide nitrogen mustard intervention group ( P=0.020, P=0.006). Hsa-miR-21-5p was transiently transfected into human umbilical cord blood mesenchymal stem cells, and the expression of has-miR-21-5p was higher than that in control group detected by qPCR ( P<0.001). The positive rate of surface protein CD9, CD63 and CD81 was 14.9%, 16.4% and 31.4%. The exosome was observed as "tea tray" or "concave hemisphere" by electron microscope. The exosome labeled by PKH-26 entered the granular cells and exerted biological effects. There was no statistically significant difference in bax mRNA expression levels between the 5 μg/mL, 10 μg/mL, and 15 μg/mL empty plasmid exosomes groups and the 60 μmol/L phosphoramide nitrogen mustard group (all P>0.05). However, the expression levels of bax mRNA in the 5 μg/mL, 10 μg/mL, and 15 μg/mL miR-21-5-p exosomes groups were lower than those in the 60 μmol/L phosphoramide nitrogen mustard group ( P=0.008, P=0.003, P<0.001). However, there was no statistically significant difference in the expression of bcl2 mRNA among the groups (all P>0.05). From the perspective of protein levels, there was no statistically significant difference in BAX protein expression between the 5 μg/mL, 10 μg/mL, and 15 μg/mL empty exosomes groups and 60 μmol/L phosphoramide nitrogen mustard group (all P>0.05), while the 5 μg/mL, 10 μg/mL, and 15 μg/mL miR-21-5-p exosomes groups showed a decrease in BAX protein expression compared with the 60 μmol/L phosphoramide nitrogen mustard group, and the differences were statistically significant (all P<0.001). However, there was no statistically significant difference in the expression of BCL2 protein among the intervention groups (all P>0.05). Conclusion:Hsa-miR-21-5p exosomes derived from human umbilical cord blood mesenchymal stem cells can effectively exert the anti-apoptotic effect.

Objective To provide direction for the improvement of quality control of hospital preparations and ensure the safety for clinical use by analyzing the hospital preparation deviations in recent three years. Methods A retrospective analysis on 59 minor hospital preparation deviations from 2017 to 2019 was conducted. Brainstorming, fishbone drawing and, Minitab software were used to analyze the root causes of deviations from five aspects: personnel, machine, materials, methods and environment. The preventive and corrective measures were implemented. The results were evaluated. Results 1 significant deviation (1.7%), 24 major deviation (40.7%), and 34 minor deviation (57.6%) were identified among the 59 casses of preparation deviation. With the implementation of preventive and corrective measures, the total number of deviations in 2018 was significantly reduced compared to that in 2017. The total number of deviations in 2019 was about the same as that in 2018. The human factors need to be focused. Conclusion The pharmaceutical preparation deviations in our hospital have been reduced. The further quality improvements for pharmaceutical preparations will be carried out by following the regulations of pharmaceutical production quality management standards and pharmaceutical production supervision and administration measures.

Objective To analyze the status quo and existing problems of radiation protection for veterinary X-ray facilitiesin Beijing-Tianjin region, China, and to provide a basis for improving radiation protection management level. Methods According to the requirements of the Technical guidelines for status quo assessment of occupational hazard of the employing unit (AQ/T 4270—2015), the study performed status quo assessment on the workplaces of veterinary X-ray facilities in 16 pet hospitals in Beijing-Tianjin region, and comprehensively analyzed the impact of the workplaces on the health of radiation workers and the public. Results In terms of personnel management, the pass rates of occupational health management post setting, personnel training, personal protective equipment, individual monitoring of occupational external exposure, notification of occupational hazards, and occupational health surveillance were 100%, 81.3%, 100%, 75%, 37.5%, and 25%, respectively. In terms of workplace management, most workplaces had reasonable layout and zoning and complete protection and emergency devices. However, there were some problems in some places, such as non-standard radiation warning signs, no notice board of occupational hazards, and no radiation protection testing. In terms of document management, all institutions had imperfect system documents. Conclusion The workplaces of veterinary X-ray facilities in 16 institutions basically meet the requirements for radiation protection, but there are also some problems, such as inadequate management of personnel and workplace radiation protection, and imperfect system documents. Institutions should strengthen the study of radiation protection knowledge. The competent authorities should strengthen supervision, formulate corresponding standards for radiation protection, enhance training, and improve the professional level of staff.

Objective To evaluate the current status of radioactive occupational hazards in the construction of some medical institutions in Tianjin, discover existingproblems and deficiencies, and provide a technical basis for radiation protection management. Methods Based on the requirements of relevant regulations and standards, we comprehensivelyevaluated the results of the pre-evaluation and control effect evaluation of 81 radioactive occupational hazards in Tianjin. Results Among the 35 proposed protective measures for pre-evaluation of radiation occupational hazards constructionprojects, the lowest compliance rate is warning signs and personal protective equipment. Among the 46 construction projects of radioactive occupational hazard control evaluation, the pass rate of protective measures was between 73.9% and 100%, with the lowest pass rate for personal protective equipment. Besides, the pass rate of radiation protectionmanagement evaluation was 95.7% ～ 100%, with the lowest pass rate for the protection management system.The initial inspectionpass rateof the equipment quality control acceptancewas 98.6%, while the pass rate was 97.1% for the relevant workplace radiation protection during the initial inspection. Conclusion The pre-evaluation completion rate of radiological occupational hazard evaluation in Tianjin medical institution construction project, unilateral and area of construction room, workplace layout and zoning, ventilation, shielding measures, personal dose management and occupational health inspection all meet the requirements of regulations and standards. However, there are still some deficiencies in the personal protective equipment, warning signs and the radiation protection management system. Therefore, the regulatory department should further strengthen the evaluation of medical institutions, especially the key supervision and management of non-conformities.

Objective:To analyze of the main influecing factors of mosic embryos during the preimplantation genetic test for chromosomal structural rearrangement (PGT-SR) to avoid the increase of risk of abortion and genetic abnormalities and to improve the diagnostic rate of mosic embryos.Methods:We used a retrospective cohort study to analyze 94 cycles of infertile patients undergoing PGT-SR and 551 cycles of intracytoplasmic sperm injection (ICSI) from January 2018 to December 2019 in the Reproductive Medical Center of the Maternal and Child Health Hospital. The relationship of mosic embryos was analyzed between the age, the number of oocytes, gonadotropin (Gn)/oocyte, the grade of blastocysts and chromosome carrier of different genders by the SPSS21.0 software.Results:In the PGT-SR cycle, single factor analysis found that mosaic embryos were related to age and sperm concentration ( P=0.02, P=0.04), but multivariate logistic regression analysis showed that age ( OR=3.41, 95% CI=1.34-8.66, P=0.01), sperm concentration ( OR=0.41, 95% CI=0.17-0.96, P=0.04) and chromosome carrier of different genders ( OR=2.21, 95% CI=1.04-4.70, P=0.04) were the main factors of embryo mosaicism. Conclusion:Female age, sperm concentration and chromosome carrier of different genders maybe affect the formation of mosaic embryos, providing theoretical basis for selective transfer of mosaic embryos.

Objective:To analyze of the main influecing factors of mosic embryos during the preimplantation genetic test for chromosomal structural rearrangement (PGT-SR) to avoid the increase of risk of abortion and genetic abnormalities and to improve the diagnostic rate of mosic embryos.Methods:We used a retrospective cohort study to analyze 94 cycles of infertile patients undergoing PGT-SR and 551 cycles of intracytoplasmic sperm injection (ICSI) from January 2018 to December 2019 in the Reproductive Medical Center of the Maternal and Child Health Hospital. The relationship of mosic embryos was analyzed between the age, the number of oocytes, gonadotropin (Gn)/oocyte, the grade of blastocysts and chromosome carrier of different genders by the SPSS21.0 software.Results:In the PGT-SR cycle, single factor analysis found that mosaic embryos were related to age and sperm concentration ( P=0.02, P=0.04), but multivariate logistic regression analysis showed that age ( OR=3.41, 95% CI=1.34-8.66, P=0.01), sperm concentration ( OR=0.41, 95% CI=0.17-0.96, P=0.04) and chromosome carrier of different genders ( OR=2.21, 95% CI=1.04-4.70, P=0.04) were the main factors of embryo mosaicism. Conclusion:Female age, sperm concentration and chromosome carrier of different genders maybe affect the formation of mosaic embryos, providing theoretical basis for selective transfer of mosaic embryos.

Objective To establish a method for simultaneous determination of chlorogenic acid, luteolin-7-O-β-D-glucoside,3,5-dicaffeoylquinic acid, linarin and pogostone in Fangshu Qingre mixture by RP-HPLC. Methods ZORBAX-SB-C₁₈ column(4.6 mm×250 mm, 5 μm) was used as the chromatographic column.The mobile phase was 0.2% formic acid water solution(A) and acetonitrile solution (B)with a gradient elution mode. The flow rate was 1.0 ml/min.The detective wavelength was 327 nm. The column temperature was 30 ℃. Results There were good linear relationships in the determination of chlorogenic acid, luteolin-7-O-β-D-glucoside, 3,5-dicaffeoylquinic acid, linarin and pogostone (r≥0.999 6), with the average recovery rate of 102.03%(1.63%), 102.38%(1.51%), 102.39%(1.23%), 103.14%(1.87%) and 104.01%(2.33%). Conclusion The method was simple and stable with a good reproducibility, which could be used as a quality control method for active compotents in Fangshu Qingre mixture.

Objective:To explore the mechanism of Golgi phosphoprotein 3 (GOLPH3) regulating the proliferation and apoptosis of endometrial cancer (EC) cells through PI3K/AKT/GSK3β signal.Methods:Human endometrial adenocarcinoma HEC-1-B cells were divided into control group, GOLPH3 group and GOLPH3 + GDC-0941 group. GOLPH3 was over-expressed by transfection of the GOLPH3 pcDNA. The PI3K inhibitor GDC-0941 was used to block the PI3K/AKT/GSK3β pathway. Cell viability, cell proliferation and apoptosis were measured by CCK-8 method, clone formation experiment and flow cytometry, respectively. The protein phosphorylation level in PI3K/AKT/GSK3β pathway was detected by Western blotting.Results:Transfection experiments were successful, and the PI3K inhibitor GDC-0941 did not affect GOLPH3 mRNA and protein expression. The relative cell viability of the control group, the GOLPH3 group and the GOLPH3 + GDC-0941 group were (100.00±4.63)%, (131.56±7.85)% and (97.85±7.36)%, and the difference among the three groups were significant ( F=7.437, P<0.001). The cell viability of the GOLPH3 group was higher than that of the control group ( P<0.001). The cell viability of the GOLPH3 + GDC-0941 group was lower than that of the GOLPH3 group ( P<0.001). The numbers of cell clones in the three groups were 46.86±3.56, 89.32±4.78 and 46.48±4.05, and the difference was significant ( F=20.437, P<0.001). GOLPH3 group had more clones than control group ( P<0.001). The number of clones formed in the GOLPH3 + GDC-0941 group was less than that in the GOLPH3 group ( P<0.001). The apoptosis rates of the three groups were (5.17±0.61)%, (2.34±0.56)% and (6.85±0.53)%, and the difference was significant ( F=11.643, P<0.001). The apoptosis rate of the GOLPH3 group was lower than that of the control group ( P<0.001), and the apoptosis rate of the GOLPH3 + GDC-0941 group was higher than that of the GOLPH3 group ( P<0.001). The phosphorylated PI3K/PI3K levels of the three groups were 1.00±0.09, 3.32±0.19 and 0.93±0.06, respectively; phosphorylated AKT/AKT levels were 1.00±0.11, 4.63±0.63 and 1.15±0.16, respectively; phosphorylated GSK3β/GSK3β levels were 1.00±0.08, 4.06±0.57 and 1.04±0.14, respectively. The differences were statistically significant ( F=12.532, P<0.001; F=16.792, P<0.001; F=15.311, P<0.001). The phosphorylation levels of each protein in the GOLPH3 group were higher than those in the control group (all P<0.001), and the GOLPH3 + GDC-0941 group were lower than the GOLPH3 group (all P<0.001). Conclusion:In EC cells, over-expression of GOLPH3 can promote cell proliferation and inhibit apoptosis by activating the PI3K/AKT/GSK3β pathway, suggesting that GOLPH3 is involved in the occurrence and development of EC.

Objective:To understand the current status of quality control of medical X-ray diagnostic equipment in Tianjin, discuss the problems existing in the process of quality control, and provide technical basis for the relevant regulatory agencies.Methods:Using stratified random sampling, a total of 401 pieces of medical X-ray diagnostic equipment distributed over 111 medical institutions in Tianjin was used for this study. X-ray multifunctional dosimeters and other types of equipment were used to perform quality control testing and evaluation under the relevant health industry standards.Results:The initial testing pass rate of these equipment was 90.0%, with the pass rates of X-ray imaging equipment lower than those of X-ray fluoroscopy equipment, digital subtraction angiography (DSA) and X-ray computed tomography (CT) ( χ2=9.203, 3.383, 5.754, P<0.05). Among the imaging equipment, the screen X-ray equipment had the lowest pass rate, followed by the breast X-ray equipment and medical digital radiography (DR). The main unqualified items were output repeatability, deviation of tube voltage indication and limit spatial resolution. DR accounted for the highest percentage of X-ray equipment that were below testing condition. The unqualified proportion of X-ray imaging equipment was higher than that of X-ray fluoroscopy equipment ( χ2=8.216, P <0.05). Among the imaging equipment, the screen X-ray equipment was seen to have the highest unaqualified proportion, followed by computed radiography (CR). Among the 401 devices in this study, the initial testing pass rates of private medical institutions was not statistically different from those of public medical institutions ( P>0.05). The initial pass rates of domestic equipment was lower than those of imported equipment ( χ2=4.576, P<0.05). Conclusions:The performance of medical X-ray diagnostic equipment in Tianjin needs to be further improved. Medical institutions should pay attention to the quality control testing of medical X-ray diagnostic equipment. The relevant regulatory agencies should strengthen oversight and management of equipment that contains unqualified items, no baseline values and below the testing conditions.