Objective To investigate the protective effect and mechanism of Astragaloside Ⅳ (AS-Ⅳ) on hypoxia/re-oxygenation induced by RAW264.7 murine macrophages.Methods The hypoxia/re- oxygenation induced RAW264.7 murine macrophages served as a model of I/R injury. The cells were divided into the control group, the model group, and the AS-Ⅳ group. The cell morphological changes of each group were observed directly under inverted microscope. Inducible nitric oxide synthase (iNOS), cluster of differentiation 206 (CD206), and peroxisome proliferator activated receptor-γ (PPAR-γ) were separately examined by RT-PCR, Western blot analysis and immunofluorescence staining.Results Compared with the model group, the numbers of cell in AS-Ⅳ group significantly increased , and the cell physiological status were much better. Compared with the model group, the iNOS immunofluorescence semi quantitative (0.62 ± 0.02 vs. 1.32 ± 0.09), the expression of mRNA (1.51 ± 0.07 vs. 3.46 ± 0.39), and protein (2.30 ± 0.14 vs. 5.16 ± 0.49) significantly reduced in AS-Ⅳ group (P<0.01). The CD206 immunofluorescence semi quantitative (1.01 ± 0.03 vs.0.61 ± 0.01), the expression of mRNA (0.91 ± 0.03 vs.0.51 ± 0.01), and protein (0.61 ± 0.04 vs.0.19 ± 0.01) significantly reduced in AS-Ⅳ group (P<0.01). Compared with the model group, the PPAR-γ immunofluorescence semi quantitative (0.60 ± 0.14 vs. 0.34 ± 0.03), mRNA (2.00 ± 0.14 vs.1.04 ± 0.03), andprotein (0.67 ± 0.05 vs.0.19 ± 0.01) significantly increased (P<0.01). Conclusions The AS-Ⅳ could attenuate ischemia-reperfusion injury by altering macrophages phenotype through upregulation PPAR-γ.

Objective To establish a mouse lethal model of influenza B virus , which will facilitate the study on the mechanism of pathogenesis , transmission of influenza B virus , development of new vaccines and drugs against influenza B virus.Methods We obtained a mouse adaptive B/Lee/1940 virus by continuously passaging it in mice for 5 cycles.The P5 virus was propagated in MDCK cells , which was used for infecting mice .The body mass and survival rate of mice were monitored during the following 14 days after infection.At the same time,the 8 gene segments (PB2, PB1, PA, HA, NA/NB, NP, M, and NS) of P0 and P5 virus were sequenced and analyzed .Results and Conclusion Virus was detected in the lungs of mice in each generation in the process of virus passaging .The body mass of mice infected with the deadly mouse adaptive virus changed dramatically .The mortality of mice was 100%, and virus was detected in mouse lungs . Sequence analysis results indicated that the amino acid mutations occurred in PB 2 and NP.A series of experiments indicated that we had established a mouse lethal model of influenza B virus .

Objective To explore self-emulsifying particle size characterization methods and compare the regularity of various methods. Methods By setting the clarity level of turbidity standard solution, with two less soluble drugs-diterpene lactone compounds Chuanhuning and dihydropyridine drug nifedipine as model drugs, 10-12 clarity level prescriptions were selected from six different ternary phase diagram. Laser particle size scanner was used to determine the particle size, and UV-visible spectrophotometry to determine its absorbance. Three methods of particle size characterization rules were compared by drawing charts. Results There was a positive correlationship among droplet particle size, absorbance and clarity grade of emulsion formed by prescription in the same phase diagram. But, there was no regularity among droplet particle size, absorbance and clarity grade of emulsion formed by prescription in different phase diagram. Conclusion The droplet particle size of emulsion formed by prescription containing the same drugs and excipients in different proportions can be compared by clarity with visual method or absorbance with UV-visible spectrophotometer.

Objective To explore the effects of the different concentrations of magnesium ions on vascular smooth muscle cell (VSMC) calcification in rats. Methods VSMCs were obtained from rat aortic, and were identified by immunocy-tochemistry. VSMCs were then randomly divided into control group, high phosphorus group and magnesium intervention group. VSMCs were cultured with 10%fetal bovine serum in control group. VSMCs were cultured with high phosphorus in high phosphorus group. VSMCs were cultured with different concentrations of magnesium chloride based on the high phos-phorus medium in magnesium intervention group (final concentrations of magnesium ions were 1, 2 and 3 mmol/L). The calci-um content and alkaline phosphatase(ALP)activity were measured after the stimulation for 7 days. The expression of Cbfα1 mRNA was detected by RT-PCR. Results Compared with control group, calcium deposits were found significantly higher in high phosphorus group and magnesium intervention group. The calcified nodules gradually reduced with the increased magnesium ion concentration in the intervention group. The calcium contents were significantly lower in the intervention groups (2 and 3 mmol/L) compared with those of high phosphorus group (P<0.05), but no difference was found between 1 mmol/L magnesium intervention group and high phosphorus group. There were no significant differences in the ALP activity and Cbfα1 mRNA expression between intervention groups (2 and 3 mmol/L) and control group (P<0.05). The ALP activity and the expression of Cbfα1 mRNA were gradually decreased with the increased magnesium ion concentration in the inter-vention group, and which were lower than those of high phosphorus group (P<0.05). Conclusion Magnesium can reduce calcification and osteoblastic transdifferentiation, which may be achieved by reducing the expression of Cbfα1 in VSMCs.

Objective:To establish a droplet digital polymerase chain reaction (ddPCR) detection method for monkeypox(mpox) virus.Methods:Specific primers and probe targeting the F3 L gene of mpox virus were used to develop a ddPCR detection method for the mpox virus′s nucleic acid. The ddPCR reaction conditions were optimized, and the specificity, sensitivity, and repeatability of ddPCR were assessed. Additional examination of clinical samples obtained from human pus swabs was performed and the ddPCR method′s ability to detect clinical samples was evaluated. Results:The result demonstrated that the mpox virus ddPCR detection method was established and the ddPCR reaction temperature and primer probe concentration has been optimized. The method had a minimal detection limit of 2 copies/μl and did not exhibit cross reaction with the remaining 8 viruses. Ten clinical samples of human skin pus swabs were tested for the mpox virus; five of the samples were tested positive by ddPCR, and four of the samples were tested positive by real time fluorescence quantitative PCR (qPCR).Conclusions:The established ddPCR for mpox virus is a fast, specific, and highly sensitive detection method, and can be used for the detection of mpox virus in clinical samples.

Objective To explore the value of the plasma miR-193a-5p level on diagnosis and monitoring the response to treatment in acute myeloid leukemia (AML). Methods Peripheral blood samples were obtained from AML patients enrolled in hematology department of the Second Hospital of Shanxi Medical University from July 2015 to December 2015, including 30 de novo AML patients, 9 patients in complete remission (CR) and 6 patients in relapse. Peripheral blood samples from 15 healthy people were randomly choosed as the health control group. Plasma miR-193a-5p expression levels were detected by using quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR). Results The plasma miR-193a-5p relative expression level of AML patients group [0.465 6 (0.103 1-5.000 2)] was obviously lower than that of health control group [0.766 1 (0.052 1-3.134 4)] (U= 122, P= 0.018 7). The plasma miR-193a-5p relative expression levels of de novo group and relapse AML group were significantly lower than those of CR group and health control group (P<0.05), and there was no significant difference between the CR group and health control group (U= 56, P= 0.511 9). No significant correlation was found between the plasma miR-193a-5p level and age, gender, blast percentage of the bone marrow, peripheral blood leukocyte count, platelet count, CD34+cells'percentage and so on. Conclusion The decreased plasma miR-193a-5p expression level can be served as a new and noninvasive biomarker for the evaluation of diagnosis and treatment in AML.

Objective To observe the expression of interferon induced protein (IP)-10 and the role of nuclear factor-kappaB (NF-κB) signaling pathway in rat peritoneal mesothelial cells (RPMCs) under the action of lipopolysaccharide (LPS).Methods RPMCs were harvested from Sprague-Dawley rat peritoneal cavity and cultured under defined in vitro conditions.The cells were exposed respectively to different concentrations of LPS (0,10,100,1 000,10 000 ng/ml) for 3 h or treated with LPS (100 ng/ml)for different time points (0,1,3,6,12,24,48 h).For observing the effect of LPS on the expression of p-p65 and p65,the RPMCs were treated with LPS (100 ng/ml) for different time points (0,15,30,60,120 min).For observing the effect of BAY11-7085 on the expression of IP-10 mRNA,the RPMCs were treated by LPS or pretreated with BAY11-7085 (5 μ mol/L) for2 h,then treated with LPS for another 3 h,respectively.Expression of IP-10 mRNA was examined by reverse transcription-polymerase chain reaction (RT-PCR).Expression of NF-κB and p-NF-κB protein was detected by Western blot.The secretion of IP-10 was determined by enzyme-linked immunosorbent assay (ELISA).Results Compared with the control group,stimulation of RPMCs with 10 ng/ml LPS resulted in a significant increase in the expression of IP-10 mRNA (P ＜0.05).1 000 ng/ml LPS has the strongest effect on IP-10 expression compared with that of 10 ng/ml and 100 ng/ml LPS.Treatment with 100 ng/ml LPS resuhed in time-dependent increase in the gene level of IP-10,with the peak at 3 h.However,after that time point,the gene level of them was gradually attenuated.Following treatment with LPS (100 ng/ml),the level of p-NF-κB began to increase at 15 min,gradually reached the peak at 1 hour,and then decreased.But the level of which at 2 h is still significant higher than that of medium control.5 μmol/L BAY11-7085 significantly decreased the up-regulation of IP-10 induced by LPS.Conclusions LPS enhanced the expression of IP-10 on RPMCs in a concentration-dependent and a time-dependent manner.LPS induced expression of IP-10 depended on the NF-κB signal transduction pathway.

Objective To study the therapeutic effect of bitter gourd saponins on salt-sensitive kidney injury induced by high salt diet and its possible mechanism.Methods 50 Sprague Dawley (SD) rats were randomly divided into normal group,model group and low-dose,middle-dose and high-dose treatment group after 10 days of adaptive feeding.Each group had 10 rats.Except the normal group,the other four groups were given high salt diet (4.0％ high salt diet) to induce salt-sensitive kidney damage in rats.The normal group and the model group were given 1.0 m/(kg · d) normal saline,and the three dosage groups of total saponins of balsam pear were given 10 mg/(kg · d),20 mg/(kg · d) and 40 mg/(kg · d) respectively.After 8 weeks of treatment,rats were sacrificed and collect the 24-hour proteinuria,creatinine.Serum creatinine,serum aldosterone,serum sodium and serum potassium were measured,and renal histopathology and the expression of podocin and nephrin were detected.Results Pathological examination of model group showed obvious glomerular sclerosis and renal interstitial fibrosis,and glomerular sclerosis in the treatment group was obviously improved by bitter gourd saponins;The systolic pressure in the model group was 170 mmHg,significantly higher than that of the normal and treatment groups,the systolic blood pressure of the treatment groups were obvious decreased when treated by bitter gourd saponins (P ＜ 0.05);Compared with normal group,serum creatinine and 24 h proteinuria / urine creatinine in model group were significantly increased (P ＜ 0.05),while creatinine clearance rate and aldosterone were significantly decreased (P ＜ 0.05),and the above indexes in bitter gourd saponins treatment group were significantly improved;Compared with the model group,the protein and mRNA expression of podocin and nephrin were significantly decreased (P ＜ 0.05),while the two indexes can be revered by bitter gourd saponins in treatment group (P ＜ 0.05).Conclusions The bitter gourd saponins can significantly improve the symptoms of salt-induced hypertensive nephropathy in rats,which may be related with the expression of podocin and nephrin in renal tissue,thereby inhibiting glomerulosclerosis and improving renal interstitial fibrosis.

Objective To investigate the imaging features and risk factors of white matter hyperin-tensities(WMH)in elderly patients without stroke symptoms in Dingxi area.Methods A total of 253 elderly WMH patients without stroke syndrome admitted to our department from January 2020 to June 2022 were recruited,and according to Fazekas classification,they were divided into a control group(92 cases)and a study group(161 cases).The general clinical data were compared between the two groups.Multivariate logistic regression analysis was used to analyze the risk fac-tors of WMH,and Spearman correlation analysis was employed to analyze the correlation between WMH severity and hypertension.Results Older age,larger proportions of male and internal carotid plaques,and higher Hcy and TG levels were observed in the study group than the control group(P＜0.05,P＜0.01).Multivariate logistic regression analysis showed that age,TG,and Hcy were risk factors for WMH in elderly patients without stroke syndrome(OR=0.564,95％CI:0.338-0.942,P=0.029;OR=1.248,95％CI:1.153-1.351,P=0.000;OR=0.046,95％CI:0.016-0.132,P=0.000).The classification of hypertension had no correlation with the severity of WMH in elderly patients without stroke syndrome.Conclusion TG,Hcy and age are independ-ent risk factors for WMH in elderly patients without stroke symptoms.

Objective:To investigate the clinical features of IgD multiple myeloma (MM) and the effect and prognosis of daratumumab-based combination therapy.Methods:The clinicopathological data of a IgD MM patient with disease progression and extramedullary infiltration treated with daratumumab in the Affiliated Hospital of Qingdao University in December 2019 were retrospectively analyzed.Results:The 74-year-old woman was diagnosed as IgD MM by bone marrow aspiration and immunofixation electrophoresis. The patient was given VD (bortezomib, dexamethasone), RD (lenalidomide, dexamethasone) and ID (ixazomib, dexamethasone) regimens. In June 2020, the patient developed multiple subcutaneous nodules, and she was assessed as progressive disease with extensive extramedullary infiltration. After treated with daratumumab-PAD (liposomal doxorubicin, bortezomib, dexamethasone) regimen, the patient's subcutaneous nodules were significantly reduced and partially disappeared, and the general condition was significantly improved. But the patient was in a cachexia state and finally died of the irregular treatment and disease progression.Conclusions:IgD MM has a low incidence and a short survival period, and there is no uniform standard treatment. The early application of daratumumab combined with proteasome inhibitors, immunomodulators, cytotoxic drugs and hematopoietic stem cell transplantation may improve the overall survival of patients.