Objective To evaluate the effect of total glucosides of paeony (TGP) on the expression of interleukin-18 (IL-18) in human HaCaT keratinocytes,and to explore the roles of extracelluar signal-regulated protein kinase1/2 (ERK1/2) and c-Jun N-terminal kinase 1/2 (JNK1/2) signaling pathways in the effect.Methods Some cultured human HaCaT keratinocytes were classified into three groups:control group treated with dimethyl sulfoxide (0.031％),TGP groups treated with 6 different concentrations (0.5,2.5,12.5,62.5,125.0 and 312.5 mg/L) of TGP respectively,inhibitor groups treated with TGP of 125 mg/L after 2-hour pretreatment with PD98059 (an ERK1/2 inhibitor) and SP600125 (a JNK1/2 inhibitor) of 10 μmol/L respectively.After additional culture for 48 hours,reverse transcription (RT)-PCR was performed to measure the mRNA expression level of IL-18,and enzymelinked immunosorbent assay (ELISA) to determine the level of IL-18 protein in the culture supematant of HaCaT cells.Some HaCaT keratinocytes were classified into two groups to be treated with TGP of 125 mg/L for 15,30 and 60 minutes with or without the pretreatment with PD98059 and SP600125 of 10 μmol/L; then,Western blot was carried out to determine the phosphorylation levels of ERK1/2 and JNK1/2 in HaCaT cells.Results The levels of IL-18 mRNA and protein in culture supernatant were significantly increased by TGP of 0.5 and 2.5 mg/L,but decreased by TGP of 62.5 and 125.0 mg/L,and TGP of 125.0 mg/L showed the strongest inhibitory effect.After treatment with TGP of 125.0 mg/L,the level of phosphorylated ERK1/2 in HaCaT cells peaked at 15 minutes (0.448 ± 0.018),decreased to 0.213 ± 0.005 at 30 minutes and 0.217 ± 0.005 at 60 minutes,with significant differences between TGP-treated and untreated cells at 15 minutes (0.448 ± 0.018 vs.0.204 ± 0.005,P＜ 0.05) but not at 30 or 60 minutes (both P ＞ 0.05).The phosphorylation level of ERK1/2 was 0.237 ± 0.010 in HaCaT cells pretreated with PD98059 prior to the treatment with TGP,significantly different from that in HaCaT cells treated with TGP only (P ＜0.01).TGP of 125.0 mg/L had no obvious effect on JNK phosphorylation,and there was no significant difference in the level of phosphorylated JNK1/2 between HaCaT cells untreated and those treated with TGP of 125.0 mg/L for different durations (all P ＞ 0.05).Conclusions TGP can inhibit the expression of IL-18 mRNA and protein in HaCaT cells,likely through the ERK1/2 signaling pathway.

ObjectiveTo investigate the effects of aconitine, mesaconitine and hypaconitine on calcium release in isolated adult rat cardiac myocytes.MethodsThe left ventricular cardiac myocytes isolated from adult Sprague-Dawley rats were perfused withacnitine, mesaconitine and hypaconitine at 0.3 μmol/L, 1μmol/L, 3 μmol/L for 12 min. The spontaneous calcium release (SCR) rate, the end-diastolic[Ca2+](F0) and the calcium transient amplitude (ΔF) were detected 4 min, 8 min and 12 min after the perfusion. 12 min after the perfusion with acnitine, mesaconitine and hypaconitine at 0.3 μmol/L, the changes of systolic dynamics and calcium transient were detected for the positive inotropic effect. Results Any of aconitine, mesaconitine and hypaconitine induced SCR, mesconitine-induced SCR rate was highest at low concentration (0.3 μmol/L), and aconitine-induced SCR rate highest at high concentration (3 μmol/L). Compared with the control, 12 min after the perfusion with acnitine, mesaconitine and hypaconitine at 3 μmol/L elevated F0 (1.459 ± 0.379, 1.585 ± 0.493, 1.213 ± 0.254vs.1.079 ± 0.108, allP<0.05) and ΔF(1.615 ± 0.455, 2.210 ± 0.756, 1.528 ± 0.422vs. 1.036 ± 0.125, allP<0.05), mesaconitine with ΔF higher than aconitine and hypaconitine. At low concentration (0.3 μmol/L), compared with control, aconitine, mesaconitine and hypaconitine increased ΔF (0.409 ± 0.127, 0.423 ± 0.107, 0.414 ± 0.118vs.0.260 ± 0.065;P<0.05 orP<0.01) and contraction amplitudes (5.464% ± 2.239%, 7.449% ± 2.548%, 5.524% ± 1.645%vs.3.428% ± 0.911%;P<0.05 orP<0.01), prolonged the time to peak of calcium transient (0.041 ± 0.016 s, 0.039 ± 0.009 s, 0.038 ± 0.011 svs.0.032 ± 0.007 s;P<0.05 or P<0.01); compared with aconitine, mesaconitine and hypaconitine decreased calcium transient time constant (0.301 ± 0.054 s, 0.324 ± 0.064 svs.0.361 ± 0.076 s;P<0.05 orP<0.01) and diastolic t50 (0.124 ± 0.035 s, 0.126 ± 0.040 svs.0.157 ± 0.056 s;P<0.05 orP<0.01).ConclusionsAconitine, mesaconitine and hypaconitine reveal the positive inotropic effects couple with the toxic effects. Increased[Ca2+]in cardiac myocytes is the key factor for the positive inotropic effects, but also the risk factor for SCR.

Objective To investigate the multidrug-resistance reversal action and mechanism of dihydroartemisin (DHA) on human colon cancer cell line HCT8/ADR. Methods The cytotoxicity of dihydroartemisin combined with doxorubicin(DOX) was determined by methyl thiazolyl tetrazolium(MTT) assay and cell apoptosis was observed by flow cytometry. Western blot assay was used to measure the autophagy. Results The combined treatment with dihydroartemisin and doxorubicin significantly enhanced the cytotoxicity in HCT8/ADR cells and effectively increased the apoptotic level. Autophagy was also induced by the combined treatment , which maybe played a crucial role in the regulation of doxorubicin-sensitization of HCT8/ADR cells. Conclusion The results indicated that dihydroartemisin can reverse multidrug resistance through increasing the doxorubicin-sensitivity of HCT8/ADR cells.

BACKGROUND: Nowadays complex bone defects have become a great challenge to orthopedists. A synergistic contribution of various growth factors and a crosstalk between their signaling pathways have been suggested as determinatives for the overall osteogenic outcome.OBJECTIVE: To develop calcium phosphate cement (CPC) incorporated with γ-polyglutamic acid/carboxymethyl chitosan (PGA/CMCS), and to evaluate its physical and chemical properties and sustained-release function. METHODS: The γ-PGA/CMCS polymer composites were prepared by graft copolymerization and spray freeze drying methods, and then loaded with recombinant human bone morphogenetic protein 2 (rhBMP-2) growth factor. CPC served as control group, and γ-PGA/CMCS-CPC containing different contents of rhBMP-2 as experimental groups. A γ-PGA/CMCS-CPC scaffold with regular blade-like crystalline structure was fabricated by injection compression molding. Before mixed with the liquid phase, the solid additives were properly mixed by wet method of CPC solid and the γ-PGA/CMCS carrier, then the pre-blended mix was freeze-dried. The setting time and compressive strength of bone cement in each group were detected, and the microstructure of the material surface was observed under scanning electron microscopy. In vitro release of rhBMP-2 was investigated. The effect of bone cement extracts on cell proliferation was determined through MTS assay.RESULTS AND CONCLUSION: γ-PGA/CMCS-CPC had the same physicochemical properties to the CPC. Initial and final setting time, compressive strength of bone cement had no significant differences among groups. The scanning electron microscope results showed that the γ-PGA/CMCS-CPC scaffold was covered by regular blade-like crystalline structure and the γ-PGA/CMCS particles were uniformly dispersed in the CPC crystals. A sustained release of rhBMP-2 was observed from the γ-PGA/CMCS-CPC. The cell experiments exhibited that the samples with regular blade-like crystalline structure had better cell response compared to CPC control groups with irregular crystalline structure. These findings indicate that γ-PGA/CMCS-CPC can maintain good physicochemical properties, and release growth factor or drug to promote bone formation.

Objective To investigate the anti-microbial activity of genistein and its mechanism. Method The anti-microbial experiment was carried out by utilizing scanning and transmission electron microscope(SEM and TEM) ,and further analyzing the respiratory metabolism and change of SDS-PAGE protein spectra. Results Genistein could inhibit several kinds of bacteria obviously.The erose structures such as rugae and bubbles were observed on the surface of cells by SEM after 24h. Moreover,with TEM,we detected the shrinkage of cytoplasm,the plasmolyses,and then the breach of wall and membrane along with the outflow of protoplasm in Staphylococcus aureus treated with genistein for 4h,14h and 24h respectively. Notablely the respiratory inhibition experiment revealed that the genistein mainly inhibits the TCA cycle of bacteria. Besides,the SDS-PAGE elucidated that the total expression of proteins was decreased in the cell treated with genistein,and especially the larger proteins were reduced with 90.1%. Conclusion Genistein showed obvious anti-microbial activity to Staphylococcus aureus. It could destroy the integrity of cell wall and membrane,prevent the respiratory metabolism and protein synthesis of the bacteria.

]. Conclusions The monoclonal-based ECLIA is a sensitive, specific, and rapid method and no radiocontamination. It can be used to detect hanum serum proinsulin in type 2 diabetes.

In order to improve transformation efficiency of dehydroepiandrosterone (DHEA) into 3beta,7alpha,15alpha-trihydroxy-5-androsten-17-one (7alpha,15alpha-diOH-DHEA) by Gibberella intermedia CA3-1, we investigated the strains breeding and their conversion process optimization. G. intermedia CA3-1 strains were treated with 0.12 mg/mL 1-methyl-3-nitro-1-nitroso-guanidin (NTG) for 30 min and chosen by 350 micromol/L minimum inhibitory concentration ketoconazole resistance marker. The high production strain named M-10 with a good genetic stability was selected and the product molar yield achieved to 70.2%, which was 20% higher than that of original strain. Under the improved conversion process with the DHEA concentration of 5 g/L, the product molar yield of the mutant M-10 reached 75.6%, which was improved by 31.3% than that of original strain.
Androstenols ; metabolism ; Biotransformation ; Dehydroepiandrosterone ; metabolism ; Gibberella ; growth & development ; metabolism ; Industrial Microbiology

Objective:To investigate the effects of Xiaoyu-Jiangzhi capsules on blood lipid, carotid artery atherosclerosis (CAA) and plaque in apolipoprotein E knockout (ApoE -/-) mice. Methods:The ApoE -/- mice were fed with high-fat food to establish carotid atherosclerosis model. The ApoE -/- mice were randomly by weight divided into model group, Atorvastatin group, low- and high-dose Xiaoyu-Jiangzhi capsules group. The C57BL/6cnc mice were used as control group and fed with normal diet. The Atorvastatin group was given atorvastatin suspension 1.3 mg/kg, low and high dose groups were given Xiaoyu-Jiangzhi capsule suspension 325 and 975 mg/kg, and the control group and model group were given equal volume of distilled water. The mice were gavaged with 0.1 ml/10 g body weight, once a day, and the weight of mice was recorded weekly. After 12 weeks of continuous intragastric administration, the blood lipid and liver /body weight index of the mice were measured. Carotid arteries were sliced to conduct oil red O staining and VG staining for the pathological analysis. Results:After 12 weeks of drug administration, the weight of mice in the high-dose group was significantly lower than the model group. The level of TC (25.92 ± 4.21 mmol/L vs. 30.39 ± 4.67 mmol/L) and LDL-C (7.97 ± 2.14 mmol/L vs. 10.26 ± 1.97 mmol/L) in the high-dose group significantly decreased ( P<0.05), the level of HDL-C in the low and high-dose group significantly increased ( P<0.05). The pathological results showed that after 12 weeks of administration, the carotid artery lipid deposition blockage rate in the Atorvastatin group and the high dose group were significantly smaller than the model group( P<0.05), and no vascular plaque has been formed. Conclusion:The Xiaoyu-Jiangzhi capsules could reduce LDL-C, increase HDL-C levels, reduce the constriction of arterial stenosis and slow down the formation process of carotid plaque.

Objective:To investigate the distribution characteristics of bacteria in urine of patients with ureteral stent crusting.Methods:Thirty-five patients who underwent ureteral stent placement at the Shandong Provincial Third Hospital, Shandong University Qilu Hospital, Jinan Central Hospital, and Jinan Jigang Hospital were selected from October, 2018 to March, 2019(the clinical study registration number is ChiCTR1800020025). The inclusion criteria were patients who had the stent intubated for 4 weeks after ureteroscopic lithotripsy, aged between 18 and 65 years. Exclusion criteria were patients with positive urine bacterial culture, severe gross hematuria, recent oral antibiotics, and patients with significant residual stones. This clinical study uses a cross-sectional study method, and those patients were divided into crusting group (n=23) and non-crusting group (n=12) according to the presence or absence of stent crusting. On the day of extubation, urine of the patients was collected for bacterial 16s DNA detection. The distribution characteristics of bacteria in urine of the two groups were analyzed using UPARSE, UCHIME and RDP calssifier. The total number of bacteria species, bacterial abundance and bacterial species with large-scale abundance in urine of the two groups were determined. The quantity of bacteria species and bacterial abundance in the urine between the two groups were compared, and the bacterial species with large-scale abundance in urine of the patients with stent crusting were identified.Results:There were no significant differences in general information such as age, body mass index, gender, affected side, type of stent tube, and stone composition between the two groups. Using 16s DNA sequencing to detect the bacteria in the urine of the two groups revealed that the number of bacterial species with abundance >1% was 11, and the number of bacterial species with abundance >0.01% was 74 in the crusting group. In the non-crusting group, the number of bacterial species with abundance >1% and >0.01% was 7 and 11, respectively. Compared with the non-crusting group, the number of bacterial species with abundance >1% in the crusting group was significantly larger ( t=5.12, P=0.000). In the crusting group, bacterial species with the top three abundance were g_Lactobacillus (23.1%), g_Bacteroides (18.8%) and g_norank_Bacteroides (17.1%). In the non-crusting group, bacterial species with the top three abundance were g_Escherichia-Shigella (32.2%), g_Enterococcus (24.9%) and g_Pseudomonas (18.2%). The three bacteria with the greatest difference between the two groups were g_ Lactobacillus ( P=0.010), g_Bacteroides ( P=0.004) and g_norank_Bacteroides ( P=0.004), respectively. Conclusion:The species and quantity of bacteria in the urine of patients with stent crusting are both significantly larger than those of patients without stent crusting. Bacteroides with larger-scale abundance in the urine of patients with stent crusting may promote the deposition of crystals on the stent wall through its structure, function and urease positive characteristics.