OBJECTIVE:To study the feasibility of purifying the effective blood-fat dropping parts from radix puerariae and fructus crataegi and to optimize the technical conditions and parameters.METHODS:Absorption capacities of four different macroporous resins on pueraria total isoflavones,crataegi total flavonoids and crataegi total triterpenic acid were compared.With adsorption quantity as index,optimization of the technics of purifying the effective blood-fat dropping parts from radix puerariae and fructus crataegi with macroporous resins were performed.RESULTS:AB-8resin showed a best adsorbability on the effective blood-fat dropping parts from radix puerariae and fructus crataegi,its optimum purification condition was the following,the concentration of the stock solution was0.13g crude drug/ml with a flow rate at0.5ml/min,and its pH value at about3.5(the same as the stock solution);80%ethanol solution was taken as the eluting solvent at an eluting flow rate of2ml/min.The purity coefficient of the purified effective portions was above85%.CONCLUSIONS:AB-8macroporous resin could be used for the purification of the effective blood-fat dropping parts from radix puerariae and fructus crataegi,the process is feasible and the reproduction of resin is easy.

Objective To investigate the mechanism of live combined Bifidobacterium and Lactobacillus tablets in acute liver failure (ALF) treatment.Methods Ten mice were injected intraperitoneally with 3.0 g/kg D-galactosamine to establish the model of ALF and treated with live combined Bifidobacterium and Lactobacillus tablets.Protein levels of Jagged1,Notch1,Notch intracellular domain (NICD),Hes5 and the mRNA expressions of Jagged1,Notch1,Hes5 were measured via Western blot and real time-polymerase chain reaction (PCR),respectively.The protein level of CD68 was detected by immunohistochemical staining method.Meanwhile,serum alanine aminotransferase (ALT),aspartate aminotransferase (AST),interleukin (IL)-10,high mobility group protein B1 (HMGB1) and plasma lipopolysaccharide (LPS) were measured.Moreover,model group and control group were also established with 10 mice each.In vitro,RAW264.7 cells were cultured with normal mice plasma,plasma of ALF mice and plasma of treated mice,respectively.Real time-PCR and Western blot were used to determine the mRNA expressions of Jagged1,Notch1,Hes5 and proteins levels of Jagged1,Notch1,NICD,Hes5.The levels of IL-10,HMGB1 and LPS in the supernatant of RAW264.7 cells were detected as well.The total significant differences among groups were compared by one way ANOVA,and q test was used to evaluate the significance of subgroup differences.Results The levels of serum ALT,AST,HMGB1,IL10,plasma LPS,and the expressions of Jagged1,Notch1,NICD,Hes5,CD68 were higher in ALF model group than control group (all P＜0.01).Compared with the ALF model group,all of these indexes could be improved in mice with live combined Bifidobacterium and Lactobacillus tablets (HMGB1:[82.6±9.7] μg/L vs [101.9±12.4] μg/L,q=6.36,P＜0.01; IL-10.:[3 183±769] pg/mL vs [4 628±842] pg/mL,q=6.79,P＜0.01; plasma LPS:[7.40±0.92] EU/mL vs [11.80±0.89] EU/mL,q=18.81,P＜0.01; Jagged1 mRNA:5.55±0.71 vs 7.63±1.41,q=7.22,P＜0.01;Jagged1 protein:0.56±0.07 vs 0.71±0.07,q=7.20,P＜0.01; Notch1 mRNA:3.66±0.67 vs 7.10±0.66,q=20.06,P＜0.01; Notch1 protein:0.38±0.08 vs 0.66±0.11,q=9.57,P＜0.01;NICD protein:0.47±0.05 vs 0.76±0.07,q=12.68,P＜0.01; Hes5 mRNA:3.94±0.68 vs 7.95± 0.71,q=22.40,P＜0.01; Hes5 protein:1.04±0.12 vs 1.20±0.07,q=5.61,P＜0.01; CD68 protein:5 180±610 vs 7 685 ±417,q=16.38,P＜0.01).And the differences were statistically significant.After RAW264.7 cells cultured with the plasma of ALF model mice,the levels of HMGB1,IL-10 and LPS in the supernatant and the expressions of Jagged1,Notch1,NICD and Hes5 in cells were increased,whereas if RAW264.7 cells were cultured with the plasma of treated mice,indexes mentioned above were significantly decreased (all P＜0.01).Conclusions Live combined Bifidobacterium and Lactobacillus tablet could prevent the occurrence and development of ALF by decreasing the plasma level of LPS,inhibiting the activation of Notch signaling pathway in macrophages and reducing the secretion of HMGB1 and IL-10.

Objective:To investigate the features of volume, distribution, grading and staging of prostate cancer(PCa)examined via whole-mount histopathology in transitional PCa.Methods:A total of 129 PCa patients undergone radical prostatectomy(RP)between July 2017 and March 2020 whose whole-mount prostate specimens were prepared after surgery were retrospectively studied.Pathological data on tumor locations, diameters and classification of the International Society of Urologic Pathology(ISUP), radiological data on regions of interest(ROI)and scores of the Prostate Imaging and Reporting Data System(PI-RADS v2)were recorded.The results of pathological whole-mount sections and prostate imaging were compared, and the characteristics and detection rates of lesions in different prostate regions were analyzed.Results:Of all 129 prostate specimens from RP, a total of 213 PCa lesions were detected through whole-mount histopathology.There were 21(9.9%)lesions involving both the peripheral zone(PZ)and the transition zone(TZ), with an average diameter of(2.82±0.71)cm.Of all lesions, 85(39.9%)involved PZ and 107(50.2%)involved TZ, with an average diameter of(1.36±0.81)cm and of(1.60±0.94)cm, respectively.The percentage of lesions involving TZ was higher than that lesions involving PZ, with larger diameters( P<0.05). Of 64 patients with complete MRI data, 105 PCa lesions were detected histopathologically by using whole mount sections, while 75 PCa lesions were detected by MRI, with a statistical difference( P<0.05). For lesions≥1.0 cm or lesions with an ISUP grade group≥2, the detection rate of MRI was lower in TZ lesions( P<0.05). Conclusions:PCa lesions within TZ account for a large proportion and have a relatively large tumor dimeter.PCa lesions within TZ are more likely to be missed in clinical examinations and on MRI, and clinicians should pay close attention during diagnosis and treatment.

OBJECTIVETo investigate the role of the Notch signaling pathway, and the underlying mechanism, in development of acute liver failure (ALF) in a mouse model.

METHODSFor in vivo analysis of the role of Notch signaling in ALF, a mouse model of ALF was generated by intraperitoneal injection of 3.0 g/kg D-galactosamine. Histological specimens were stained by hematoxylin-eosin, and then studied microscopically.Expression level of Jaggedl, Notchl, NICD, and Hes5 was measured by western blotting (for protein) and real time-PCR (for mRNA). The level of CD68 protein was detected by immunohistochemical staining. Serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), IL-10, high mobility group box 1 (HMGB1) chromatin protein, and lipopolysaccharide (LPS) were measured by standard methods. For in vitro analysis of the molecular mechanism, the RAW264.7 macrophage cell line was cultured with LPS in the absence or presence of the Notch inhibitor DAPT, and the intracellular levels of Notch1, NICD, and Hes5 were measured by western blotting and real time-PCR and the extracellular levels of IL-10 and HMGB1 were detected in the supematant.

RESULTSCompared with unmodeled (normal control) mice, the ALF modeled mice showed higher levels of serum ALT (848.40+/-94.83 U/L vs. 38.99+/-9.63 U/L), AST (911.49+/-67.65 U/L vs. 55.28+/-7.50 U/L), HMGB1 (101.91+/-12.43 µg/L vs. 20.73+/-5.37 µg/L), 1L-10 (4 627.88+/-842.45 pg/mL vs. 1 064.92+/-238.46 pg/mL) and LPS (11.80+/-0.89 EU/mL vs. 0.58+/-0.12 EU/mL), as well as higher expression of Jagged1 (mRNA: 7.63+/-1.41 vs. 1.00+/-0.00; protein: 0.71+/-0.07 vs. 0.34+/-0.07), Notch1 (mRNA: 7.10+/-0.66 vs. 1.00+/-0.00; protein: 0.66+/-0.11 vs. 0.27+/-0.08), NICD (protein: 0.76+/-0.08 vs. 0.27+/-0.08), Hes5 (mRNA: 7.95+/-0.71 vs. 1.00+/-0.00; protein: 1.20+/-0.07 vs. 0.76+/-0.07), and CD68 (protein: 7 685.05+/-417.34 vs. 2 294.01+/-392.93) (all P<0.01). In vitro, LPS increased the extracellular levels of HMGB1 (7.44+/-0.63 vs. 0.21+/-0.05), IL-10 (315.19+/-79.13 vs. 59.19+/-23.30) and the intracellular expression of Notch1 (mRNA: 6.49+/-0.73 vs. 1.00+/-0.00), NICD (protein: 0.65+/-0.10 vs. 0.23+/-0.07), and Hes5 (mRNA: 7.30+/-0.85 vs. 1.00+/-0.00; protein: 0.96+/-0.10 vs. 0.54+/-0.07) (all P<0.01). DAPT treatment led to a decrease above the index serum levels of HMGB1 (6.22+/-0.71) and IL-10 (252.06+/-57.63), and of expression of Notch 1 (mRNA: 3.20+/-0.68), NICD (protein: 0.42+/-0.05), and Hes5 (mRNA: 4.72+/-0.67; protein: 0.84+/-0.09) (P<0.01 or <0.05).

CONCLUSIONThe Notch signaling pathway may plan an important role in the development of ALF upon activation of the pathway in macrophages by LPS and leading to promoted secretion of HMGB 1 and IL-10, with a greater effect on the former.

Alanine Transaminase ; Animals ; Aspartate Aminotransferases ; Disease Models, Animal ; Galactosamine ; HMGB1 Protein ; Lipopolysaccharides ; Liver Failure, Acute ; Mice ; RAW 264.7 Cells ; RNA, Messenger ; Receptors, Notch ; metabolism ; Signal Transduction

Objective To measure the transaction costs of compact county medical alliance and propose counter-measures.Methods Based on Williamson-Zhang Wuchang analysis paradigm,it established the measure matrix of transaction cost of compact county medical alliance.Through the typical sampling method,the medical community of D city in Haixi Prefecture,Qinghai Province was selected as a research example to carry out an empirical study.Results The total transaction cost of the compact county medical community was 6 292 500.00 yuan,the transac-tion cost of the leading hospital was 6 234 300.00 yuan,accounting for 99.07％,and the special investment of the leading hospital reached to 5 887 100.00 yuan,accounting for 94.43％.The total transaction cost of the 298 referred patients was 308 400.00 yuan,and the average transaction cost per patient was 1 035.05 yuan.Conclusion The transaction costs of compact county medical alliance was generally high,most of which was borne by the leading hospital,and the patients also had to bear the corresponding transaction costs.In the next stage,the transaction costs should be dealt with from three aspects,including recognizing the secrecy and universality of the transaction costs,establishing economical system supply and establishing a sound compensation mechanism.

Objective To measure the transaction costs of compact county medical alliance and propose counter-measures.Methods Based on Williamson-Zhang Wuchang analysis paradigm,it established the measure matrix of transaction cost of compact county medical alliance.Through the typical sampling method,the medical community of D city in Haixi Prefecture,Qinghai Province was selected as a research example to carry out an empirical study.Results The total transaction cost of the compact county medical community was 6 292 500.00 yuan,the transac-tion cost of the leading hospital was 6 234 300.00 yuan,accounting for 99.07％,and the special investment of the leading hospital reached to 5 887 100.00 yuan,accounting for 94.43％.The total transaction cost of the 298 referred patients was 308 400.00 yuan,and the average transaction cost per patient was 1 035.05 yuan.Conclusion The transaction costs of compact county medical alliance was generally high,most of which was borne by the leading hospital,and the patients also had to bear the corresponding transaction costs.In the next stage,the transaction costs should be dealt with from three aspects,including recognizing the secrecy and universality of the transaction costs,establishing economical system supply and establishing a sound compensation mechanism.

Objective To measure the transaction costs of compact county medical alliance and propose counter-measures.Methods Based on Williamson-Zhang Wuchang analysis paradigm,it established the measure matrix of transaction cost of compact county medical alliance.Through the typical sampling method,the medical community of D city in Haixi Prefecture,Qinghai Province was selected as a research example to carry out an empirical study.Results The total transaction cost of the compact county medical community was 6 292 500.00 yuan,the transac-tion cost of the leading hospital was 6 234 300.00 yuan,accounting for 99.07％,and the special investment of the leading hospital reached to 5 887 100.00 yuan,accounting for 94.43％.The total transaction cost of the 298 referred patients was 308 400.00 yuan,and the average transaction cost per patient was 1 035.05 yuan.Conclusion The transaction costs of compact county medical alliance was generally high,most of which was borne by the leading hospital,and the patients also had to bear the corresponding transaction costs.In the next stage,the transaction costs should be dealt with from three aspects,including recognizing the secrecy and universality of the transaction costs,establishing economical system supply and establishing a sound compensation mechanism.

Objective To measure the transaction costs of compact county medical alliance and propose counter-measures.Methods Based on Williamson-Zhang Wuchang analysis paradigm,it established the measure matrix of transaction cost of compact county medical alliance.Through the typical sampling method,the medical community of D city in Haixi Prefecture,Qinghai Province was selected as a research example to carry out an empirical study.Results The total transaction cost of the compact county medical community was 6 292 500.00 yuan,the transac-tion cost of the leading hospital was 6 234 300.00 yuan,accounting for 99.07％,and the special investment of the leading hospital reached to 5 887 100.00 yuan,accounting for 94.43％.The total transaction cost of the 298 referred patients was 308 400.00 yuan,and the average transaction cost per patient was 1 035.05 yuan.Conclusion The transaction costs of compact county medical alliance was generally high,most of which was borne by the leading hospital,and the patients also had to bear the corresponding transaction costs.In the next stage,the transaction costs should be dealt with from three aspects,including recognizing the secrecy and universality of the transaction costs,establishing economical system supply and establishing a sound compensation mechanism.

Objective To measure the transaction costs of compact county medical alliance and propose counter-measures.Methods Based on Williamson-Zhang Wuchang analysis paradigm,it established the measure matrix of transaction cost of compact county medical alliance.Through the typical sampling method,the medical community of D city in Haixi Prefecture,Qinghai Province was selected as a research example to carry out an empirical study.Results The total transaction cost of the compact county medical community was 6 292 500.00 yuan,the transac-tion cost of the leading hospital was 6 234 300.00 yuan,accounting for 99.07％,and the special investment of the leading hospital reached to 5 887 100.00 yuan,accounting for 94.43％.The total transaction cost of the 298 referred patients was 308 400.00 yuan,and the average transaction cost per patient was 1 035.05 yuan.Conclusion The transaction costs of compact county medical alliance was generally high,most of which was borne by the leading hospital,and the patients also had to bear the corresponding transaction costs.In the next stage,the transaction costs should be dealt with from three aspects,including recognizing the secrecy and universality of the transaction costs,establishing economical system supply and establishing a sound compensation mechanism.

Objective To measure the transaction costs of compact county medical alliance and propose counter-measures.Methods Based on Williamson-Zhang Wuchang analysis paradigm,it established the measure matrix of transaction cost of compact county medical alliance.Through the typical sampling method,the medical community of D city in Haixi Prefecture,Qinghai Province was selected as a research example to carry out an empirical study.Results The total transaction cost of the compact county medical community was 6 292 500.00 yuan,the transac-tion cost of the leading hospital was 6 234 300.00 yuan,accounting for 99.07％,and the special investment of the leading hospital reached to 5 887 100.00 yuan,accounting for 94.43％.The total transaction cost of the 298 referred patients was 308 400.00 yuan,and the average transaction cost per patient was 1 035.05 yuan.Conclusion The transaction costs of compact county medical alliance was generally high,most of which was borne by the leading hospital,and the patients also had to bear the corresponding transaction costs.In the next stage,the transaction costs should be dealt with from three aspects,including recognizing the secrecy and universality of the transaction costs,establishing economical system supply and establishing a sound compensation mechanism.