ObjectiveTo clarify the scientific validity of in vivo pharmacokinetic determination of the whole drug composition in Shenbai nanosuspension in rats， and to provide methodological guidance and theoretical basis for the in vivo study of multi-component complex system of traditional Chinese medicine（TCM） preparations. MethodThe concentration of the overall components， mainly total saponins and total polysaccharides in Shenbai decoction and Shenbai nanosuspension， was determined in rat plasma at different times by area under the absorbance-wavelength curve method（AUAWC）， and the concentration of individual ginsenoside Rg₁ was determined by high performance liquid chromatography（HPLC）， and the methodology was verified. The pharmacokinetic parameters of the whole component were compared with those of ginsenoside Rg₁ to evaluate the in vivo operational characteristics of the two preparations. ResultThe methodological investigations of AUAWC and HPLC were in accordance with the requirements. AUAWC analysis showed that the overall components in both the decoction group and the nanosuspension group showed a one-compartment model， with half-life（t_1/2） of 2.43 h and 2.04 h， respectively. The relative bioavailability of Shenbai nanosuspension was 138.99%. HPLC assay showed that ginsenoside Rg₁ in the decoction group and the nanosuspension group showed a two-compartment model， with distribution half-life（t_1/2α） of 0.13 h and 2.55 h， and elimination half-life（t_1/2β） were 14.28 h and 3.85 h， respectively. The relative bioavailability of Shenbai nanosuspension was 127.49%. Compared with Shenbai decoction， the time to peak（t_max）， peak concentration（C_max） and area under the drug-time curve（AUC） of the overall components and ginsenoside Rg₁ in Shenbai nanosuspension were increased. ConclusionThe established AUAWC can be used for the pharmacokinetic study of the overall components of TCM preparations， which is complementary to the results of individual components measured by HPLC， and can provide useful reference for the in vivo study of new dosage forms of TCM.

Objective To study the correlation between semen quality and bacterial infection in men with abnormal fer-tility,and provide clinical basis for guiding the reproductive health of men with abnormal fertility.Methods 200 male semen samples with abnormal fertility were collected,and then separated and cultured for 48 hours.According to the culture results,they were divided into three groups:the non-pathogenic group,the pathogenic group,and the sterile group.The bacterial resist-ance analysis of the pathogenic group was conducted,and the semen quality between each group was compared.Results After 48 hours of isolation and cultivation,200 semen samples had been tested,non-pathogenic bacteria was detected in 163 semen samples,accounting for 81.5% ;pathogenic bacteria was detected in 33 semen samples,accounting for 16.5% ;and bacteria was not detected in 4 semen samples,accounting for 2.0% .The top three strains of pathogenic bacteria in 33 cases were Escherichia coli,Streptococcus agalactiae,and Enterococcus faecalis,with drug resistance rates of 80.0% ,87.5% ,and 100.0% ,respec-tively.Conclusion The detection rate of bacterial culture in semen of men with abnormal fertility is relatively high,and patho-genic bacteria can affect semen quality.

Objective To investigate the effect of exosomes derived from Echinococcus multilocularis on macrophage polarization after treatment for different durations and concentrations. Methods A total of 60 BALB/c mice were used for modeling, among which 4 mice were selected to observe the growth of abdominal lesions on 7.0T MRI. The mice for modeling were dissected, and the protoscoleces was taken from the abdominal lesion and cultured in vitro ; ultracentrifugation was used to extract the exosomes from the supernatant, and transmission electron microscopy and Western blotting were used for the characterization of exosomes. The macrophages without exosome treatment were established as control group, and the macrophages co-cultured with different concentrations of exosomes derived from Echinococcus multilocularis were established as experimental group (10 μg/mL group and 50 μg/mL group) and were cultured for 48 and 72 hours. The morphological changes of macrophages were observed under a microscope, and flow cytometry and ELISA were used to observe polarization state. A one-way analysis of variance was used for comparison of normally distributed continuous data between multiple groups, and the least significant difference t -test was used for further comparison between two groups. Results The results of 7.0T MRI showed the formation of diffuse lesions with different sizes in the abdominal cavity of mice, and the exosomes derived from Echinococcus multilocularis were approximately 100 nm in diameter and were cup-shaped or saucer-shaped, with the positive expression of the surface markers CD9, TSG101, and CD63. After co-culture, most of the cells in the experimental group were elongated with an irregular and polygonal shape. Flow cytometry showed that after 48 hours of co-culture, the positive rates of CD16/32, CD206, and CD369 in the control group were 99.53%±0.06%, 90.27%±0.21%, and 2.40%±0.20%, respectively; compared with the control group, except that the 10 μg/mL exosome group had a significant reduction in the positive rate of CD369 (0.80%±0.00%) ( P < 0.05), all the other groups had a significant increase in the positive rates of CD16/32, CD206, and CD369 (all P < 0.000 1); after 72 hours of co-culture, the positive rates of CD16/32, CD206, and CD369 in the control group were 99.67%±0.06%, 85.47%±0.55%, and 6.60%±0.20%, respectively, and compared with the control group, the experimental group had significant increases in the positive rates of CD16/32, CD206, and CD369 (all P < 0.05). ELISA showed that after 48 hours of co-culture, the levels of IL-6 and TNFα in the control group were 58.53±15.52 pg/mL and 320.70±5.30 pg/mL, respectively, and when the exosome concentration was 50 μg/mL, the level of IL-6 in the experimental group was 98.81±15.55 pg/mL, which was higher than that in the control group ( P < 0.05); after 72 hours of co-culture, the levels of IL-6 and TNFα in the control group were 76.22±9.68 pg/mL and 323.90±87.37 pg/mL, respectively, and when the exosome concentration was 10 μg/mL, the level of TNFα was 164.20±14.17 pg/mL, which was significantly lower than that in the control group ( P < 0.05); when the exosome concentration was 50 μg/mL, the level of IL-6 was 99.52±8.35 pg/mL, which was significantly higher than that in the control group ( P < 0.05). Conclusion Exosomes derived from Echinococcus multilocularis can regulate macrophage polarization and induce M2-like polarization of macrophages after co-culture at a concentration of 10 μg /mL for 72 hours, and further studies are needed to clarify the specific method.

As the most commonly used antipyretic and analgesic drug,paracetamol(PA)coexists with neuro-transmitter dopamine(DA)in real biological samples.Their simultaneous determination is extremely important for human health,but they also interfere with each other.In order to improve the conductivity,adsorption affinity,sensitivity,and selectivity of TiO2-based electrochemical sensor,N-doped carbon@-TiO2 double-shelled hollow sphere(H-C/N@TiO2)is designed and synthesized by simple alcoholic and hydrothermal method,using polystyrene sphere(PS)as a template.Meanwhile,TiO2 hollow spheres(H-TiO2)or N-doped carbon hollow spheres(H-C/N)are also prepared by the same method.H-C/N@TiO2 has good conductivity,charge separation,and the highly enhanced and stable current responses for the detection of PA and DA.The detection limit and linear range are 50.0 nmol/L and 0.3-50 μmol/L for PA,40.0 nmol/L and 0.3-50 μmol/L for DA,respectively,which are better than those of carbon-based sen-sors.Moreover,this electrochemical sensor,with high selectivity,strong anti-interference,high reli-ability,and long time durability,can be used for the simultaneous detection of PA and DA in human blood serum and saliva.The high electrochemical performance of H-C/N@TiO2 is attributed to the multi-functional combination of different layers,because of good conductivity,absorption and electrons transfer ability from in-situ N-doped carbon and electrocatalytic activity from TiO2.

Objective : Tnfrsf11 a Cre Rosa26 yfp reporter gene mice were prepared to determine the efficiency of Cre-mediated recombination using flow cytometry in different tissue-resident macrophages. Methods : TheTnfrsf11 a Cre Rosa26 yfp reporter mice were generated by crossingTnfrsf11 a Cre mice withRosa26 yfp mice and identified by PCR.Brain microglia, liver macrophages, kidney macrophages, alveolar macrophages and spleen macrophages were separated from adultTnfrsf11 a Cre Rosa26 yfp reporter mouse and yellow fluorescent protein(YFP) labeling efficiency was analyzed by flow cytometry. Results : YFP expression percentage was about 91.27% of brain microglia inTnfrsf11 a Cre Rosa26 yfp reporter mice, but liver, spleen, and alveolar macrophages were inefficiently targeted(63.60%,69.66%,32.76%). Conclusion Tnfrsf11 a Cre mice were qualified as conditional knockout model mice of brain microglia andTnfrsf11 a Cre Rosa26 yfp reporter mice can be used as a tool for conditional knockout of brain microgliain vivo.

Objective:To investigate the effect of miR-425-5p on glucagon-like peptide-1(GLP-1) secretion in intestinal L cells induced by lipopolysaccharide(LPS), and to explore its mechanism.Methods:GLUTag cells of intestinal L cell line were incubated with LPS to determine the levels of miR-425-5p and GLP-1. Cell viability was determined by MTT assay, and cell apoptosis was detected by flow cytometry. Quantitative real time-PCR and western blot were performed to determine the expressions of miR-425-5p, phosphatase and tensin homology(PTEN), proglucagon, and GLP-1. Activity of Wnt/β-catenin signaling pathway was determined by detecting TOP/FOP ratio. Interaction among miR-425-5p, PTEN, and β-catenin was analyzed using luciferase activity assay and chromatin immunoprecipitation(ChIP)assay.Results:In GLUTag cells, with the elevation of LPS concentration, the expression of miR-425-5p and the apoptosis rate were increased, while the level of active GLP-1 and the cell viability were decreased. MiR-425-5p was involved in the regulation of LPS on GLP-1 secretion and intestinal L cell viability. Inhibition of miR-425-5p reduced the mRNA expression of proglucagon and the TOP/FOP ratio, increased PTEN protein level, and inhibited cell viability. In LPS-treated GLUTag cells, miR-425-5p increased the level of β-catenin by targeting PTEN, and β-catenin acted as a cis-acting element to induce the transcription of proglucagon and promote the secretion of GLP-1.Conclusion:In LPS-induced intestinal L cells, miR-425-5p promotes the expression of GLP-1 by targeting PTEN to modulate β-catenin.

Objective:To quantify the kinematic changes in the gait cycle among youths with chronic, nonspecific low back pain (CNLBP).Methods:A Qualisys 3D optical motion capture system was used to document the gait kinematics of 11 youths with CNLBP and 11 healthy counterparts. Gait profile scores and movement analysis profiles were then calculated.Results:The young CNLBP subjects exhibited gait abnormalities, as reflected by the significant difference of the gait variable the range of motion of the dominant knee between the CNLBP and healthy groups [(5.66±1.83)° vs (3.64±1.13)°]. Gait profile scores and movement analysis profiles are applicable to evaluating them.Conclusions:There manifested gait abnormalities in youth CNLBP patients. Gait profile scores and movement analysis profiles are applicable to evaluating them.

Objective By taking usage of bioinformatics screening methods,this medical research aimed at exploring how the miRNAs in endothelial progenitor cell exosomes relate to the regulation of angiogenesis.Methods miRNAs in endothelial progenitor cells exosomes and angiogenesis-related miRNA was intersected from the existing database to obtain the candidates of miRNA molecules related to angiogenesis in endothelial progenitor exosomes.Results 160 and 50 miRNAs in endothelial progenitor cell candidates were obtained through experimental data analysis and literature searching respectively.600 candidates of angiogenesis-related miRNAs were obtained through literature searching;the top 20 with the highest frequency were selected out.Finally,9 miRNA candidates (miR-126,miR-21,miR-221,miR-92a,miR-199a,miR-210,miR-214,miR-155,miR-146a) that may be highly expressed in endothelial progenitor exosomes and associated with angiogenesis were obtained for the following research.Conclusions Based on data analysis and literature searching,bioinformatics could screen out the target miRNAs for follow-up studies easily and reliably,it is worthy to be widely applied and popularized.

Objective To study the safety and effectiveness of polymer-free sirolimus-eluting Nano stent in patients with unstable angina pectoris. Methods Three hundred and twenty-one patients with unstable angina pectoris were divided into Nano stent group(group A,n=157)and Endeavor resolute stent group(group B,n=164). The cardiovascular events were compared postoperative 12 months. The minimal intima cavity area and mini-mum bracket section area and neointimal area were compared postoperative 12 months by intravascular unltrasound (IVUS). Results There were 7 cases of cardiovascular events in group A and 6 in group B postoperative 12 months(P=0.727)and 2 patients in group A and 3 in group B were re-implanted stent because of restenosis post-operative 12 months(P=0.672). The neointimal area were(0.31 ± 0.11 mm2)in group A and(0.29 ± 0.12 mm2) in group B postoperative 12 months(P = 0.985). The minimal intima cavity area(P = 0.921)and the minimum bracket section area(P=0.934)were narrower postoperative 12 months than immediately after the operation in two groups. Conclusion With less cardiovascular events and being safe and reliable,the clinical effect of polymer-free sirolimus-eluting Nano stent implantation is similar to that of Endeavor resolute stent implantation.

Objective To investigate the click and tone burst evoked auditory brainstem responses (ABR)in normal Wistar rat,and to establish the standards of ABR testing method,and to provide a reference for studies rat audition.Methods Fifteen male Wistar rats(30 ears)were used in this sutdy.The latency and amplitude of ABR e-voked by click and TB at 80,50 and 20 dB SPL were measured.Results The occurrence rate of wave Ⅱand Ⅳat low levels(20 dB SPL)was nearly the same according to the amplitude.The cABR (dB peSPL)threshold was 21.83± 4.45 and tbABR (dB SPL)thresholds were 2.02±0.09,2.88±0.16,3.77±0.25,4.69±0.29,and 5.78±0.41, respectively.80 dB stimulus evoked cABR (peSPL)wave I,I b,II,III,IV and V latency (ms)were 1.76±0.12, 2.13±0.11,2.67±0.16,3.49±0.28,4.39±0.29,and 5.45±0.41,respectively.tbABR (SPL)of wave I,Ib, II,III,IV and V latency (ms)at 4 kHz were 2.02±0.09,2.88±0.16,3.77±0.25,4.69±0.29,and 5.78± 0.41,respectively.At 8 kHz they were 1.76±0.07,2.28±0.10,2.63±0.16,3.49±0.21,4.44±0.28,and 5.48±0.43;while at 12 kHz were1.76±0.08,2.24±0.12,2.61±0.25,3.53±0.25,4.46±0.32,and 5.52± 0.45;at 16 kHz were 1.79±0.10,2.25±0.12,2.70±0.18,3.62±0.27,4.52±0.37,and 5.61±0.49;at 24 kHz were 1.75±0.09,2.27±0.11,2.67±0.16,3.60±0.27,4.52±0.38,and 5.60±0.51;at 32 kHz were 1.77±0.10,2.24±0.12,2.64±0.20,3.59±0.34,4.52±0.40,and 5.61±0.52,respectively.Conclusion Wave Ⅳ was the best wave to determine threshold of click and tone burst evoked auditory brainstem response in rat.