Objective:To establish the method validation of microbial limit tests for hospital paste preparations, including com-pound salicylic acid paste, zinc oxide paste and compound pine tar paste. Methods:According to the microbial limit test described in China pharmacopoeia 2010 edition, the method validation of count of bacteria, fungi and yeasts and tests for specified microorganisms was established. Results:Medium dilution method could be used in bacteria, fungi and yeasts count and specified microorganisms ex-amination for compound salicylic acid paste and zinc oxide paste. For compound pine tar paste, bacteria, fungi and yeasts count and the pseudomonas aeruginosa examination could use medium dilution method, while the staphylococcus aureus examination should employ membrane filtration method. Conclusion:The methods of microbial limit tests for the three hospital paste preparations are established, which can be used to control the quality of hospital preparations effectively.

Objective To investigate protection by immunization with recombinant Ferritin vaccine of Echinococcus granulosus against protoscolices.Methods ICR mice were randomized into 3groups of 12 mice in each.The mice in group A and B were immunized three times with an interval of two weeks and those in group C did nothing.The animals in all the 3 groups were challenged with 1100 protoscolices intraperitoneally on the 8th week.Serum samples were collected before each inoculation and challenge injection.Seven months later, all the mice were killed and examinated for hydatid cysts.Result The number of cysts was significantly lower in the group A than in group B and C (P＜0.05).The levels of protection afforded were found to be 73% and 85%, respectively.Meanwhile,the number of cysts was markedly lower in group B than in group C(P＜0.05).The rate of protection afforded was 42%.Conclusion Recombinant Ferritin vaccine of Echinococcus granulosus shows partial immune protection.Therefore, it might be a suitable candidate for cocktail vaccine study in the future.

Objective:To establish the microbial limit test methods for thirteen kinds of ointments. Methods:The microbial limit of 13 kinds of ointments was respectively determined by the routine method, culture medium dilution method and membrane filtration method. Results:The recovery of the tested bacteria in the samples was above 70% by the different methods. Conclusion:The micro-bial limit test methods for thirteen kinds of ointments are stablished, which may be used in the quality control.

AIM: To investigate the expression of tissue factor(TF) induced by oxidized high density lipoprotein(oxHDL) in human umbilical vein cell line,ECV304,and the related mechanisms.METHODS: Four main groups were designed: the negative,the positive(ECV304 with histamine),the HDL group and the oxHDL group.Quantitative real-time polymerase chain reaction(RQ-PCR) and Western blotting were used to detect the expression level of TF.The specific inhibitors of MAPKs,SP600125(c-jun terminal NH2 kinase,JNK),SB203580(p38 MAP kinase,p38 MAPK),PD98059(extracellular signal-regulated kinase,ERK1/2) were used to investigate the underlying mechanisms.RESULTS: The TF expression in normal ECV304 cell line was not detected.Histamine administration resulted in a significant expression of TF in ECV304 cell line,with strongest effect after 1 h co-incubation at concentration of 1?10-5 mol/L histamine(about 4.8-fold higher expression of TF compared with that of 1?10-9 mol/L histamine).Expression level of TF was detected after stimulated with oxHDL in dose-and time-dependent manners.The highest expression of TF mRNA was found at 20 mg/L oxHDL and 6 h co-incubation,with 1.8-fold and 5.3-fold increase in TF expression,respectively,compared with that at 10 mg/L oxHDL and 2 h co-incubation.20 mg/L oxHDL also caused an apparent augmentation of TF protein expression,about 1.5-fold higher compared with that stimulated by 40 mg/L oxHDL.HDL co-incubation did not cause a detectable expression of TF protein.The mRNA levels of TF in ECV304 cell line induced by oxHDL were decreased by 95.0%,81.0%,87.0%,respectively(all P

Objective:To explore the block-based charging method for centralized dispensing of neonatal drugs in pharmacy intravenous admixture service (PIVAS), analyze its effect on drug savings and inpatient drug cost, so as to provide the reference for the appropriate charging method of neonatal drugs.Methods:According to the balance quantity and amount of neonatal intravenous drugs that were centrally allocated by the PIVAS of our hospital, refer to the doctor′s orders, the dosage per dose as well as the number of patients per dose were analyzed, then the drug types and plans for block-based charging were formulated. Before and after the implementation of the plan, the monthly average drug balance quantity and amount, the average number of drug charges for the neonates, the average daily drug cost, and the adverse events of related drugs were used as the indicators to be investigated to clarify the implementation effect of the block-based charging mothod.Results:Fourteen medicines were charged by block-based, including 4 antibiotics, 2 ordinary infusion preparations, and 8 parenteral nutrition solution preparations. The monthly average drug balance quantity was reduced from 5 047±541 to 1 856±225, and the monthly average balance amount was reduced from 65 811±10 265 yuan to 20 659±6 002 yuan. The average drug dosage for children in the trial drug was significantly reduced with a decrease range of 39.2% to 90.1%. Both the inpatient daily drug cost of neonatus and the daily average antibacterial drug cost was decreased. During the centralized dispensing of neonatal drugs, no related adverse drug events occurred.Conclusions:The block-based charging method of centralized drug distribution can improve the utilization rate of drugs, reduce drug waste, reduce the cost of inpatient medicines the financial burden on children′s families, which is worthy of further promotion and implementation.

Objective:To evaluate the effect of oropharyngeal aspiration on reducing ventilator-associated pneumonia,in order to provide a basis for clinical operation and practice.Methods:CNKI, Wanfang, China Biology Medicine Disc, VIP, PubMed, Embase, Cochrane Library, Web of Science and CINAHL databases were searched for relevant studies on the effects of oropharyngeal attraction on ventilators associated pneumonia. At the same time, the included references were screened, and the search time limit was from the establishment of the database to January 1, 2023. The quality of the included literature was strictly evaluated and data extracted by two researchers with evidence-based training, and Meta-analysis was performed by RevMan 5.4 software.Results:The results of meta-analysis showed that oropharyngeal aspiration could reduce the incidence of ventilator-associated pneumonia ( OR=0.30, 95% CI 0.22-0.40, P<0.01) and shorten the duration of mechanical ventilation ( MD=-2.39, 95% CI -3.59--1.20, P<0.01) and reduced length of stay in ICU ( MD=-2.62, 95% CI -2.99--2.26, P<0.01). Conclusions:Strengthening oropharyngeal aspiration can reduce the incidence of ventilator-associated pneumonia in patients with mechanical ventilation. In clinical practice, oropharyngeal aspiration should be performed when the position changes, and continuous aspiration can be performed if conditions are met.

Objective To explore the value of peripheral blood circulating DNA in predicting the efficacy and prognosis of immunotherapy for advanced non-small cell lung cancer.Method A retrospective study was conducted on 78 NSCLC patients who were admitted to the Respiratory and Critical Care Medicine Department of the First Affiliated Hospital of Hebei North University and were treated with tirelizumab for advanced driver gene negativity from January 2021 to December 2021.After 2 cycles of immunotherapy,the efficacy was evaluated according to the Solid Tumor Efficacy Evaluation Criteria(RECIST 1.1),including complete remission,partial remission,disease stability,and disease progression.CR and PR patients were defined as the experimental group(n=48)Other patients were defined as the control group(n=30),and the ctDNA levels in peripheral blood were measured before and after treatment in both groups.ROC curves were used to analyze the predictive value of periph-eral blood ctDNA levels for achieving objective remission after immunotherapy.All patients were followed up and their progression free survival were calcutated.Using univariate and multivariate regression analysis identified the factors affecting the prognosis of patients after immunotherapy.Using Spearman correlation coefficient analyzed the correlation between ctDNA levels and PFS.Kalplan Meier survival curve were used for survival analysis.Result The peripheral blood ctDNA levels before and after treatment in the experimental group were(4.47±1.21)ng/μL and(2.65±1.14)ng/μL,respectively(t=7.559,P<0.001),while those in the control group were(4.54±1.15)ng/mL and(4.29±1.57)ng/μL,respectively(t=0.699,P=0.487).There was no statistically significant difference in peripheral blood ctDNA levels between the two groups before treatment(t=-0.25,P=0.801).The peripheral blood ctDNA levels in the experimental group decreased compared to the control group after treatment(t=-5.35,P<0.001).The ROC curve analysis showed that the area under the curve for predicting objective remission after immunotherapy based on peripheral blood ctDNA levels was 0.819,with a sensitivity of 81.3%and specificity of 80%.Peripheral blood ctDNA levels were negatively correlated with progression free survival(r=-0.784,P=0.000).Single factor COX regression was used to analyze the clinical and pathological characteristics and ctDNA levels of enrolled patients,and the results showed that the maximum tumor diameter was greater than 5 cm(HR=0.501,95%CI:6.731～35.567)Tumor stage IV(HR=0.392,95%CI:0.227～0.677),treatment approach(HR=15.473,95%CI:6.731～35.567),and ctDNA levels(HR=4.657,95%CI:3.182～6.555)are all influencing factors for PFS in advanced NSCLC patients after immunotherapy.Multiple factor analysis was conducted on the appeal indicators with statistical differences,and the results showed that treatment approach(HR=2.981,95%CI:1.019～8.722)and peripheral blood ctDNA levels(HR=3.918,95%CI:2.619～5.861)It is an independent influencing factor of PFS in advanced NSCLC patients.The Kalplan Meier survival curve was used for analysis,and the results showed that the median PFS of the treatment effective group was 8.4 months,while the median PFS of the control group was 5.4 months.(χ2=49.277,P=0.000).Conclusion Immunotherapy combined with chemotherapy can enhance the ability to kill tumor cells,and peripheral blood ctDNA levels can evaluate the efficacy and prognosis of immunotherapy,which can be used to guide immunotherapy in advanced NSCLC patients.

Objective:To observe the effect of bone morphogenetic protein 4 (BMP4) on the proliferation and migration of human retinal microvascular endothelial cells (hRMEC) under oxidative stress.Methods:The hRMEC cultured in vitro were divided into control group, 4-hydroxynonenal (HNE) treatment group (4-HNE group), 4-HNE+BMP4 group (BMP4 group). Cell culture medium of 4-HNE treatment group was added with 10 μmmol/L 4-HNE; cell culture of BMP4 group was cultured with 10 μmmol/L 4-HNE, and after stimulation for 6 h, 100 ng/ml recombinant human BMP4 was added. The effects of 4-HNE and BMP4 on hRMEC viability was detected by thiazole blue colorimetric method. The effects of 4-HNE and BMP4 on cell migration was determined by cell scratch test. The relative expression of BMP4 mRNA in the cells of the control group and 4-HNE treatment group and the mRNA expression of the control group, the fibronectin (FN) of BMP4 group, laminin (Laminin), α-smooth muscle contractile protein (α-SMA), and collagen type Ⅰ (Collagen Ⅰ), vascular endothelial growth factor (VEGF), and connective tissue growth factor (CTGF) were detected by real-time quantitative polymerase chain reaction (qRT-PCR). Western blot was used to detect the relative expression of BMP4 protein in the control group and 4-HNE group. The control group and 4-HNE group were compared by t test. Results:Compared with the control group, cell viability ( t=12.73, 16.26, P=0.000 2, <0.000 1), cell migration rate ( t=28.17, 37.48, P<0.000 1, <0.000 1) in 4-HNE group and BMP4 group were significantly increased, and the difference was statistically significant; the relative expression of BMP4 mRNA and protein in the 4-HNE group was significantly increased, and the difference was statistically significant ( t=16.36, 69.35, P=0.000 1, <0.000 1). The qRT-PCR test results showed that compared with the control group, the relative expression of VEGF, FN, Laminin, α-SMA, Collagen Ⅰ, and CTGF mRNA in the cells of the BMP4 group was significantly increased, and the difference was statistically significant ( t=10.61, 17.00, 14.85, 7.78, 12.02, 10.61, P=0.0004, <0.000 1, 0.000 1, 0.001 5, 0.000 1, 0.000 4). Conclusion:BMP4 can induce the proliferation and migration of hRMEC; it can also regulate the expression of angiogenesis factors and fibrosis-related factors in hRMEC.

Background Arsenic can be toxic to human by triggering oxidative stress, which is companied by epigenetic modifications. Objective To investigate the modification of N⁶-methyladenosine (m⁶A) in human embryonic lung fibroblasts (HELF) during oxidative stress induced by sodium arsenite (NaAsO₂). Methods HELF cells were treated by designed concentrations of NaAsO₂ (0, 2.5, 5, 10, and 20 μmol·L⁻¹) for 48 h. Cell viability was detected by 3-(4,5-dimethylthia zol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfopheny)-2H-tetrazolium (MTS) method; the activities of total superoxide dismutase (T-SOD) and glutathione peroxidase (GSH-Px) as well as the content of malondialdehyde (MDA) were detected with corresponding kits; the level of m⁶A methylation in total RNA was detected by enzyme-linked immunosorbent assay; the mRNA expressions of m⁶A modified enzymes were detected by real-time fluorescence quantitative PCR, including methyltransferase-like 3 (METTL3), methyltransferase-like 14 (METTL14), Wilms' tumor 1-associated protein (WTAP), fat mass and obesity-associated protein (FTO), alkB family of Fe(II)/α-ketoglutarate-dependent dioxygenases 5 (ALKBH5), YTH domain containing protein 2 (YTHDC2), YTH domain family protein 2 (YTHDF2), and YTH domain family protein 3 (YTHDF3); the protein expressions of METTL3, FTO, YTHDC2, YTHDF3, and nuclear factor erythroid 2-related factor 2 (NRF2) were detected by Western blotting. The enrichment of m⁶A in NRF2 mRNA was detected by RNA methylated immunoprecipitation combined with real-time fluorescence quantitative PCR (MeRIP-qPCR). Results After the 0, 2.5, 5, 10, and 20 μmol·L⁻¹ NaAsO₂ treatment, the MTS results showed that compared with the control group, the cell viability of the 20 μmol·L⁻¹ group decreased to 84% (P<0.05). The colorimetry results showed that compared with the control group, the activities of T-SOD in the 10 and 20 μmol·L⁻¹ groups decreased (P<0.05); the activities of GSH-Px in the 2.5 and 10 μmol·L⁻¹ groups decreased (P<0.05); the contents of MDA in the 10 and 20 μmol·L⁻¹ groups increased. The results of enzyme-linked immunosorbent assay showed that the overall m⁶A methylation levels in the 0, 2.5, 5, 10, and 20 μmol·L⁻¹ groups were (0.193 ± 0.023)%, (0.247 ± 0.021)%, (0.253 ± 0.006)%, (0.233 ± 0.006)%, and (0.262 ± 0.010)%, respectively, and compared with the control group, the m⁶A methylation levels in all the NaAsO₂ treated groups increased (P<0.05). The real-time fluorescence quantitative PCR results showed that compared with the control group, the mRNA relative expression level of METTL3 decreased in the 2.5, 10, and 20 μmol·L⁻¹ groups (P<0.05); the mRNA relative expression level of FTO decreased in the 20 μmol·L⁻¹ group; the mRNA relative expression level of YTHDC2 increased in the 10 and 20 μmol·L⁻¹ groups (P<0.05); the mRNA relative expression level of YTHDF3 increased in the 2.5, 10, and 20 μmol·L⁻¹ groups (P<0.05). The Western blotting results showed that compared with the control group, the relative protein expression of METTL3 decreased in the 10 and 20 μmol·L⁻¹ groups; the relative protein expression of FTO decreased in the 5 and 20 μmol·L⁻¹ groups; the relative protein expression of YTHDC2 decreased in the 20 μmol·L⁻¹ group (P<0.05); the relative nuclear protein expression of NRF2 decreased in the 10 and 20 μmol·L⁻¹ groups (P<0.05). The MeRIP-qPCR results showed that m⁶A enrichment was significantly increased in the 20 μmol·L⁻¹ NaAsO₂ exposure group compared with the control group (P<0.05). After over-expression of FTO, the mRNA and protein relative expression levels of FTO and the relative expression level of nuclear protein of NRF2 in the FTO group were higher than those in the control group (P<0.05); the mRNA and protein relative expression levels of FTO in the NaAsO₂ + FTO group and the nuclear protein expression level of NRF2 were higher than those in the NaAsO₂ group (P<0.05). Conclusion In the process of oxidative stress induced by NaAsO₂, m⁶A methylation level, m⁶A modified enzymes, m⁶A modification of NRF2 mRNA, and NRF2 expression could change in HELF cells.