BACKGROUND: Myogenetic seed cells are the fundamental for construction of tissue-engineered smooth muscle, and optimizing the amplification of seed cells is the key in further clinical application. OBJECTIVE: To analyze the myogenetic potential of rabbit adipose-derived stromal cells induced with the combination of MyoD, transforming growth factor β1 and 5-azacytidine, and to investigate a new way to induce cells. METHODS: The rabbit abdominal fats were isolated and then the adipose-derived stromal cells were separated by col agenase digestion method for myogenetic induction with different methods: 5-azacytidine induction group, MyoD+transforming growth factor β1 induction group, 5-azacytidine+MyoD+transforming growth factor β1 combination induction group. The blank control group was set. The morphological characteristics of cells were observed at 1, 4, 8, 12, 16, 20, 24 and 28 days after induction, and the col agen type Ⅲ level were detected with 4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, flow cytometry and immunohistochemistry. RESULTS AND CONCLUSION: Compared with the other groups, the cel activity was higher in the combination induction group and reached peak at 16 days after induction, the number and volume of myotubes were increased at 20 days with regular arrangement, and desmin showed strong expression. Cel cycle showed the ratio of adipose-derived stromal cells in the DNA replication phase was increased in the combination induction group, the ratio of cells in the clearance period was decreased; the level of col agen type Ⅲ was increased significantly, and the difference was significant. The results indicate that 5-azacytidine combined with multiple factors can promote the differentiation of adipose-derived stromal cells into myoblasts with significant cel proliferation, which is considered as the ideal method to in vitro induction of myoblasts.

Objective To develop a rapid ultra high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/ MS) for determination of 11 sedative-hypnotics in human plasma. Methods The plasma samples were extracted with dichloromethane:n-hexane:acetoacetate (5∶4∶1). The separation was performed on a Waters ACQUITY UPLC HSS T3 column (2.1 mm×100 mm,1.8 μm) using the mobile phase composed of acetonitrile-0.1% ammonium solution at a flow rate of 0.2 mL?min-1 in gradient elution mode. The detection was performed on a triple quadrupole tandem mass spectrometer by multiple reaction monitoring (MRM) via electrospray ionization (ESI) source in positive mode. Results Good linear relation was obtained over the investigated concentration range, with all correlation coefficients higher than 0.99. The limit of detection was 200 pg?mL-1 for dexzopiclone,and 10- 20 pg?mL-1 for other sedative-hypnotics. The intra- and inter-day RSD of 11 sedative-hypnotics were no more than 15.0%. The recoveries were in the range of 72.3%-108.0%, and the matrix effects were approximately 0.86- 1. 12. Conclusion The method is rapid, sensitive and reliable, and suitable for the simultaneous determination of 11 sedative-hypnotics in human plasma.

OBJECTIVE:To investigate the compatible stability of Calcium folinate for injection mixed with Glucose injection and Sodium chloride injection. METHODS:Referring to clinical common concentration,each 3 Calcium folinate for injection (each injection was equal to calcium folinate 100 mg)were respectively mixed with Glucose injection 250 mL or Sodium chloride injection 250 mL. At room temperature,under light or dark condition,the appearance of mixtures,pH value and the number of in-soluble particles were investigated 0,1,2,3,4,6,8,12,24,36,48 h. The contents of calcium folinate in mixtures were deter-mined by HPLC. RESULTS:Under above condition,the color of the mixtures had no change,and no gas,precipitation and turbid-ity was found;there was no evident change in pH values(RSD<2%,n=11). 0 h after mixing,there was large number of parti-cles≥10 μm in mixtures,but the number of particle was decreased as time;within 48 h,the number of particles ≥10 μm and ≥25 μm in mixtures were all in line with the standard of Chinese Pharmacopeia(2015 edition). Under the protection from light con-dition,relative contents of calcium folinate in mixtures had no significant change(RSD<2%,n=11). Under light condition,rela-tive contents of calcium folinate in mixtures decreased significantly,decreasing to 94.5%(mixed with Glucose injection) and 88.4%(mixed with Sodium chloride injection). CONCLUSIONS:Calcium folinate for injection is more stable in Glucose injec-tion,and the stability of compatibility can be affected by light conditions. After mixed with Glucose injection and Sodium chloride injection,Calcium folinate for injection should be kept away from light and used as soon as possible

Objective:To study the stability of compound Kushen mixed liquid to provide reliable evidence for safe clinical drug application.Methods:Compound Kushen injection was mixed respectively with 0.9% sodium chloride injection and 5% glucose injection,and then placed under the different conditions for 0,1,2,4,8,12,24 and 48 h.HPLC was conducted to determine the content changes of four components in compound Kushen mixed liquid,and the appearance,pH and insoluble particles were observed as well.Results:The mixed liquid of compound Kushen with 0.9% sodium chloride injection was stable in 48 h without the influence of light and temperature.However,the mixed liquid of compound Kushen with 5% glucose injection had poorer stability with storage time shorter than 12 h at room temperature and 2 h at high temperature.Conclusion:The stability of the mixed liquid of compound Kushen is closely related to the pH value of solvent.0.9% Sodium chloride injection is recommended as the solvent,and the mixed liquid should be used up in 48 h.

Objective To evaluate the correlation and difference of reversed phase high performance liquid chromatography (RP-HPLC) and fluorescence polarization immunoassay (FPIA) on determining serum concentration of carbamazepine.Methods Fifty serum samples were collected,both RP-HPLC and FPIA methods were employed to determine the concentration of carbamazepine.The results were analyzed by paired t test,Bland-Altman and Deming regression methods,respectively.Results The results of measuring 50 samples by the two methods showed that FPIA datas were significantly higher than RP-HPLC datas,and there was statistically significant difference(P<0.05) and poorer consistency between two methods;There was good correlation between carbamazepine concentrations determined by the two methods.Deming regression equation was CFPIA=1.195 3 CRP-HPLC-0.144 0,and Pearson correlation coefficient was 0.968 5.Conclusion Clinicians should pay more attention to the difference of carbamazepine concentration determination by different methods when carbamazepine individualized dosage regimen was adjusted according to therapeutic drug monitoring.

OBJECTIVE:To investigate the compatible stability of Xiao'aiping injection combined with 3 kinds of common in-jections. METHODS:Referring to package inserts,Xiao'aiping injection 40 mL was compatible with 5% Glucose injection,10%Glucose injection or 0.9% Sodium chloride injection 160 mL,respectively. At room temperature(about 25 ℃)and high tempera-ture(40 ℃),the appearance of mixtures were observed at 0,1,2,4,8,12,24,48 h;pH value and the number of insoluble particles were detected. The contents of tenacissoside A and tenacissoside Ⅰ in mixtures were determined by HPLC. RESULTS:Un-der above condition,the mixtures were brownish yellow liquid within 48 h after Xiao'aiping injection was compatible with 5%Glucose injection or 10% Glucose injection;24 h after mixed with 0.9% Sodium chloride injection,the mixture changed from brownish yellow to reddish brown,but no precipitation was found. The pH value of mixtures had no significant change(RSD<1%,n=8). The number of particles ≥25 μm was in line with the requirements of Chinese Pharmacopeia(2015 edition). For-ty-eight hours after mixing,the number of particles ≥10 μm in the mixtures exceeded the pharmacopoeia limits. Within 48 h after mixing,the relative contents of tenacissoside A and tenacissoside I in mixtures had no significant change(RSD<2%,n=8). CON-CLUSIONS:The mixture should be used up within 24 h after Xiao'aiping injection combined with 5% Glucose injection,10%Glucose injection or 0.9% Sodium chloride injection.

OBJECTIVE:To develop a method for rapid,accurate and simultaneous determination of dexzopiclone,zolpidam and zaleplon in human plasma. METHODS:After the plasma sample was processed by liquid-liquid extraction,UPLC-MS/MS was established to detect plasma sample with carbamazepine as internal standard. The separation was performed on a Waters ACQUITY UPLC HSS T3 column with mobile phase composed of 0.1% ammonium solution-acetonitrile (gradient elution) at flow rate of 0.2 ml/min. The column temperature was set at 40 ℃,and sample size was 5 μl. The detection was performed by multiple reaction monitoring (MRM) via electrospray ionization (ESI) source in positive mode. The mass transition ion-pairs were as follows:m/z 308.2→263.1(zolpidem),m/z 389.3→245.0(dexzopiclone),m/z 306.2→236.1(zaleplon),m/z 237.3→151.2(internal standard). RE-SULTS:The linear range of zolpidem,dexzopiclone and zaleplon were 0.02-20.00,0.50-20.00,0.02-20.00 ng/ml,respectively (r=0.990 1,0.996 8,0.991 7). LLOQ of them were 0.02,0.50,0.02 ng/ml,and detection limit were 0.01,0.20,0.01 ng/ml. RS-Ds of intra-day and inter-day were all lower than 15% ;extraction recovery were 88.9% -106.5% ;matrix effect were 94.8%-106.3%. CONCLUSIONS:The method is simple,rapid,sensitive and specific,and it can be used for simultaneous deter-mination of zaleplon,zolpidem and dexzopiclone in human plasma.

Objective:To study the effects and mechanisms of activation of human lung fibroblasts (MRC-5) by exosomal RNA hsa _ circ _ 0006357 (circEZH2) derived from non-small cell lung cancer.Methods:Western blot was used to detect exosome molecular markers, reverse transcription-quantitative real-time polymerase chain reaction (RT-qPCR) and cell invasion assays to detect the effect of non-small cell lung cancer-derived exosomes on MRC-5 activation. A circRNA microarray analysis was performed in serum exosomes from patients with non-small cell lung cancer (collected at Ningbo University People's Hospital, September 2023), and levels of circEZH2 were measured in serum exosomes from non-small cell lung cancer by RT-qPCR analysis. The effects of circEZH2 on MRC-5 activation were explored using wound healing assays, Transwell assays, RT-qPCR, cellular immunofluorescence, and western blot. The regulatory effect of circEZH2 on miR-495-3p/TPD52 axis and NF-κB pathway through dual-luciferase assay, immunofluorescence, and western blot.Results:Exosomes derived from non-small cell lung cancer cells were shown to promote MRC-5 cell invasiveness, the number of cells invaded in co-culture with exosomes derived from normal human bronchial epithelial cells was (42±5), and the number of cells invaded in co-culture with exosomes derived from non-small cell lung cancer cells (SPC-A1, H1299, A549 cells) was (246±7), (89±4), (69±14), expression of markers of fibroblast activation (α-SMA, FAP), and cytokines (IL-6, IL-8, P＜0.05). CircEZH2 expression was significantly upregulated in the serum exosomes of non-small cell lung cancer ( P＜0.01). Alternatively, co-culture of exosomes derived from non-small cell lung cancer cells with MRC-5 cells promoted circEZH2 expression ( P＜0.05). Functionally, overexpression of circEZH2 promoted MRC-5 cell migration and invasion [the cell migration rates were (30.81±2.54)% and (60.29±8.34)%, respectively, and the cell invasion numbers were (48.00±13.58) and (115.00±9.50), respectively, P＜0.05]. RT-qPCR, western blot, and immunofluorescence assays demonstrated a significant increase in the expression of the pro-inflammatory genes IL-6 and IL-8 in MRC-5 cells as well as the activation markers α-SMA and FAP in fibroblasts ( P＜0.05) following the expression of circEZH2. Mechanistically, circEZH2 may function as ceRNA to regulate miR-495-3p, promote the expression of TPD52, and activate the NF-κB pathway to promote the activation of MRC-5. Conclusion:Exosomal circEZH2, derived from non-small cell lung cancer, may promote the activation of fibroblasts by activating the NF-κB pathway through the miR-495-3p/TPD52 axis.

Objective:To study the effects and mechanisms of activation of human lung fibroblasts (MRC-5) by exosomal RNA hsa _ circ _ 0006357 (circEZH2) derived from non-small cell lung cancer.Methods:Western blot was used to detect exosome molecular markers, reverse transcription-quantitative real-time polymerase chain reaction (RT-qPCR) and cell invasion assays to detect the effect of non-small cell lung cancer-derived exosomes on MRC-5 activation. A circRNA microarray analysis was performed in serum exosomes from patients with non-small cell lung cancer (collected at Ningbo University People's Hospital, September 2023), and levels of circEZH2 were measured in serum exosomes from non-small cell lung cancer by RT-qPCR analysis. The effects of circEZH2 on MRC-5 activation were explored using wound healing assays, Transwell assays, RT-qPCR, cellular immunofluorescence, and western blot. The regulatory effect of circEZH2 on miR-495-3p/TPD52 axis and NF-κB pathway through dual-luciferase assay, immunofluorescence, and western blot.Results:Exosomes derived from non-small cell lung cancer cells were shown to promote MRC-5 cell invasiveness, the number of cells invaded in co-culture with exosomes derived from normal human bronchial epithelial cells was (42±5), and the number of cells invaded in co-culture with exosomes derived from non-small cell lung cancer cells (SPC-A1, H1299, A549 cells) was (246±7), (89±4), (69±14), expression of markers of fibroblast activation (α-SMA, FAP), and cytokines (IL-6, IL-8, P＜0.05). CircEZH2 expression was significantly upregulated in the serum exosomes of non-small cell lung cancer ( P＜0.01). Alternatively, co-culture of exosomes derived from non-small cell lung cancer cells with MRC-5 cells promoted circEZH2 expression ( P＜0.05). Functionally, overexpression of circEZH2 promoted MRC-5 cell migration and invasion [the cell migration rates were (30.81±2.54)% and (60.29±8.34)%, respectively, and the cell invasion numbers were (48.00±13.58) and (115.00±9.50), respectively, P＜0.05]. RT-qPCR, western blot, and immunofluorescence assays demonstrated a significant increase in the expression of the pro-inflammatory genes IL-6 and IL-8 in MRC-5 cells as well as the activation markers α-SMA and FAP in fibroblasts ( P＜0.05) following the expression of circEZH2. Mechanistically, circEZH2 may function as ceRNA to regulate miR-495-3p, promote the expression of TPD52, and activate the NF-κB pathway to promote the activation of MRC-5. Conclusion:Exosomal circEZH2, derived from non-small cell lung cancer, may promote the activation of fibroblasts by activating the NF-κB pathway through the miR-495-3p/TPD52 axis.

Objective: To understand the status quo and influencing factors of overweight and obesity in preschool children, and to provide scientific theoretical reference for the prevention and control of overweight and obesity in preschool children in Urumqi. Methods: Stratified cluster sampling method was adopted to select 1 897 preschool children from 10 kindergartens in Urumqi from October to December in 2021 to understand the status quo of overweight and obesity of preschool children by measuring their height and weight. The influencing factors were collected by questionnaire survey, including Chinese preschoolers eating behavior questionnaire, Chinese preschooler s caregivers feeding behavior scale, 3-6 year old children s home nurture environment scale, and characteristics and influencing factors of physical activity among preschool children. Results: The prevalence of overweight and obesity in preschool children was 31.21% (592), including 19.50% (370) overweight and 11.70% (222) obesity.Childhood overweight and obesity detection rates varied significantly by age, sex, child dietary habits, father BMI, maternal BMI, and maternal pre pregnancy BMI ( χ 2=19.63,28.75,9.45,18.21,18.45,19.36, P <0.05). Multivariate Logistic regression analysis showed that gender, children s eating habits, paternal BMI, pregnancy BMI, satiety responsiveness, external eating, initiative eating, weight concerns, behavior restricted feeding, physical activity and family physical activity environment were the influencing factors of overweight and obesity in preschool children( OR =0.52,1.43,1.51,1.44,0.69,0.74,1.35,1.71,0.81,0.96,1.10, P < 0.05 ). Conclusion Overweight and obesity in preschool children are popular in Urumqi. Education, diet control and scientific exercise should be strengthened to prevent childhood overweight and obesity.