Objective To investigate the expression of SP 100 protein in ATRA-treated NB4 cells and its effect on pro-liferation in NB4 cells.Methods Q-PCR was employed to measure the expression of SP 100 mRNA;Western blot was used to detect the expression of SP 100 protein; Immunofluorescence was adopted to determine the location of SP100;Cell viability was analyzed by CCK 8;Flow cytometry was used for cell cycle analysis .Results ATRA may induce the expression of mRNA and protein of SP 100.ATRA changes the location of SP100 from a micro-punctate pattern into a punctate nuclear pattern in NB 4 cells.SP100-shRNA promotes the proliferation of NB 4 cells and in-creased the cells in G2/M phase.Conclusions The expression of SP100 was significantly increased in ATRA-treated NB4 cells, and SP100 may be involved in the regulation of proliferation activity of NB 4 cells.

AIM:To construct recombinant adenovirus vector carrying human miR-133a and study its expression in human mesenchymal stem cells(hMSCs).METHODS:The PCR product containing miR-133a was amplified from human genomic DNA and inserted into the adenoviral shuttle vector pAdTrack-CMV.Then the recombinant shuttle plasmid linearized by pmeⅠwas cotransformed into competent E.coli.BJ5183 with the adenoviral backbone plasmid pAdEasy-1 to generate the recombinant adenovirus vector rAd-mir-133a.rAd-mir-133a was then packaged and amplified in human embryonic kidney 293(HEK293) cells.The purified rAd-miR-133a was used to infect the hMSCs and the expression of miR-133a was detected by non-quantitative RT-PCR and real-time PCR.RESULTS:The recombinant adenovirus shuttle vector pAdTrack-CMV-miR-133a was constructed and verified by restriction endonuclease analysis and DNA sequence analysis.rAd-miR-133a was successfully packaged and amplified in HEK293 cells.The transcriptions of primary miR-133a and mature miR-133a were over-expressed in the hMSCs infected with rAd-miR-133a.CONCLUSION:The recombinant adenovirus vector carrying human miR-133a is successfully constructed,which lay a foundation for miR-133a function study.

Objective:To comprehensively evaluate and compare the early warning system's ability to predict the risk of maternal disease in China, such as the British National Intensive Care Audit and Research Center (ICNARC)-Obstetric Early Warning Score (OEWS) system, the National Maternal Safety Early Warning Standard (MEWC) recommended by the National Coalition for Maternal Safety, the Irish Maternity Early Warning System (IMEWS) proposed in the clinical practice guidelines issued by the head of health services at the Royal Medical College of Ireland.Methods:A total of 872 pregnant women who were admitted to the Women′s Hospital of Nanjing Medical University, Nanjing Maternity and Child Health Care Hospital were prospectively observed from January 2019 to December 2019. The data during hospitalization were collected to calculate the scores of the three obstetric early warning scoring systems, to compare and evaluate the prediction and identification ability of three early warning systems for critically ill pregnant women.Results:A total of 872 cases were included in this study, of which 167 were critically ill, accounting for 19.2%, and 705 were non-critically ill, accounting for 80.8%. The three obstetric early warning systems (OEWS,MEWC,IMEWS) in critical group scored 4(2, 6),3(0, 6),5(3, 6), and non-critical group scored 1(0, 1), 0(0, 0), 2(0, 3), respectively. The critical group scored significantly higher than the non-critical group ( P<0.001). The AUC of OEWS was 0.961 (95% CI 0.948-0.975, P<0.001) and that of MEWC was 0.803 (95% CI 0.757 - 0.850, P<0.001). The AUC of IMEWS was 0.853 (95% CI 0.817-0.888, P<0.001). No obstetric critical illness occurred when OEWS was 0. The positive likelihood ratio was 8.208 6 when OEWS≥6. Conclusions:Three kinds of early warning systems have good predictive ability for the occurrence of critical obstetric diseases in China. Among the three warning systems, OEWS has better predictive ability than the other two, and has better hierarchical ability. However, due to the differences in population, environment and conditions in different regions, each obstetric medical unit can improve OEWS according to its own conditions and design an obstetric early warning system that meets its own clinical needs.

ObjectiveTo explore the possible mechanism of Yupingfeng Granules （玉屏风散） in preventing and treating chronic obstructive pulmonary disease （COPD） from the perspective of “lung-gut axis”. MethodsThirty-two male Wistar rats were randomly divided into normal group，model group， roxithromycin group and Yupingfeng Granules group， with 8 rats in each group. Except for the normal group， the rat model of COPD was prepared by intratracheal instillation of lipopolysaccharide （LPS） combined with smoking for 12 weeks. Since the fifth week of modeling，the roxithromycin group and the Yupingfeng Granules group were given 31.5 mg/（kg·d） and 1.575 g/（kg·d） of corresponding drugs respectively by gavage，and normal group and model group were given 10 ml/（kg·d） physiolo-gical saline. Sample was collected 24 hours after the last administration. The pathological changes of lung tissue were observed using HE staining； Ultrahigh performance liquid chromatography quadrupole time of flight mass spectrometry （UHPLC-QTOFMS） was used to detect the differential metabolites in alveolar lavage fluid （BALF） in all groups but roxithromycin group；16S rDNA sequencing technology was used to detect the changes of intestinal flora， and the association analysis was conducted between the differential metabolites and the differential flora. ResultsCompared with the normal group， the model group showed an increase in goblet cells in the small bronchial wall， disappearance of the smooth muscle layer of the bronchial wall， and infiltration of inflammatory cells； compared with the model group， roxithromycin group showed slight alveolar interstital edema， and obviously reduced inflammatory cell， while no obvious alveolar interstital edema was observed in the Yupingfeng Granules group， showing a small amout of inflammatory cell infiltration. The results of the BALF differential metabolite screening showed that compared with the normal group， 12 substances were upregulated and 19 substances were downregulated in the model group； compared with the model group， 37 substances in the Yupingfeng Granules group were upregulated and 43 substances were downregulated KEGG analysis yielded a total of 2 metabolic pathways， glycerophospholipid metabolism， and unsaturated fatty acid biosynthetic metabolism； compared with the model group， choline， acetylcholine， glycerol-3-phosphate， glycerophosphate choline， palmitic acid， and arachidonic acid showed an upward trend， while stearic acid and docosahexaenoic acid showed a downward trend in Yupingfeng Granules group （P＜0.05）. The results of the intestinal flora showed that， there are 80 different species between the normal group and the model group， and 65 different species between the model group and Yupingfeng Granules group. Among the top 5 species with relative abundance levels，compared with the model group， the level of Prevotella_9，Ruminococcaceae_UCG-005，Ruminiclostridium_6 increase，and Lactobacillus，Bacteroides decrease（P＜0.05）.The results of the correlation analysis showed that， in the normal and model groups， arachidonic acid was negatively correlated with Oribacterium（r=-0.753，P＜0.01）； in the Yupingfeng Granules group and model group， stearic acid and Bacteroides（r=0.788）， Mycobacterium（r=0.826），［Eubacterium］_Ruminantium_Group（r=0.770） was positively correlated（P＜0.01）， Arachidic acid was negatively correlated with Roseiarcus（r=-0.779）， glycerol-3-phosphate was negatively correlated with Desulfovibrio（r=-0.758）， Arachidonic acid was negatively correlated with Oribacterium（r=-0.753）， and Palmitic acid was negatively correlated with Pseudolabs（r=-0.750，P＜0.01）. ConclusionYupingfeng Granules can affect the metabolism of BALF and the flora structure of intestinal microorganisms， and regulating the balance of “lung-gut axis” may be one of the mechanisms of Yupingfeng Granules in treatment of COPD.

Objective:To explore the regulatory effects of cow milk with the addition of breast milk equivalent dose of somatostatin (SST) and motilin (MTL) on food allergy and food intolerance.Methods:Young Brown Norway (BN) rats were divided into 5 groups and fed with pure breast milk(breast milk group), cow milk(cow milk group), cow milk added with SST(SST group), cow milk added with MTL(MTL group) and cow milk added with both SST+ MTL(SST+ MTL group). Allergic irritation was enhanced with skin smear at the same time.Clinical damages were quantified weekly.Levels of serous total Immunoglobulin E (IgE) and fecal calprotectin (FC) were detected by the enzyme-linked immunosorbent assay (ELISA). Gastric emptying ratio and intestinal propulsion ratio were measured by method of dextran blue.Results:In breast milk group, cow milk group, SST group, MTL group and SST + MTL group, the levels of IgE were (45.75±5.05) μg/L, (580.42±45.24) μg/L, (290.38±22.88) μg/L, (424.26±22.17) μg/L, (209.49±17.59) μg/L, respectively; FC level were (149.07±24.78) μg/g, (458.85±33.81) μg/g, (343.63±34.97) μg/g, (407.79±29.62) μg/g, (296.83±28.77) μg/g, respectively; the total score of clinical damage were (0.50±0.61) scores, (9.37±1.04) scores, (6.83±1.49) scores, (7.00±1.14) scores, (5.37±1.19) scores, respectively.The cow milk group had the highest scores of clinical damages.Compared with the cow milk group, the clinical damage score, IgE and FC of the SST, MTL and SST+ MTL groups had significantly lower levels, and there was significant difference among them (all P<0.01). The general status of the SST + MTL group was most similar to the breast milk group.The gastric emptying rate of MTL group was the closest to that of breast milk group [(92.52±6.27)% vs.(100.00±9.70)%, P<0.05]. There were obvious diarrhea and fast small intestinal propulsion in cow milk group, the small intestinal propulsion ratio in breast milk group was (39.32±2.61)%, and (71.96±4.43) % in cow milk group, the difference was statistically significant between the 2 groups ( P<0.01). The intestinal motility of SST+ MTL group was decreased, but it just prevented diarrhea caused by milk allergy, the small intestine propulsion ratio in SST+ MTL group was (38.90±2.65)% vs.breast milk group (39.32±2.61)%( P>0.05). Conclusions:The cow milk added with SST and MTL decreased allergic reaction and increased food tolerance in gastrointestinal tract, which was more similar to breast milk.SST was beneficial to relieving allergic immune reaction, MTL contributed to improving the gastrointestinal tolerance of cow milk.The combination of SST and MTL may achieve an antagonistic and balanced mechanism on gastrointestinal regulation, which could synergistically improve the gastrointestinal tolerance of cow milk.