Objective To optimize the purification technology of total flavonoids from the Tripterospermi Chinensis Herba by macroporous resin. Methods The purification abilities of six kinds of macroporous adsorption resins for total flavonoids from Tripterospermi Chinensis Herba were studied with the adsorption and desorption rates as the index by static adsorption and desorption experiments. The column liquid concentration, adsorption rate, and the loaded amount were studied by using dynamic adsorption experiments. The optimal desorption technology was examined by orthogonal experiment. Results AB-8 macroporous resin was found to have good adsorption and desorption effects. The optimal purification conditions were as follows:the column liquid concentration was 5.285 mg/mL, adsorption rate was 2 BV/h;the loaded amount was 17.62 mg/mL. The sample was eluted by 10%ethanol with 4 BV and 50%ethanol of 5 BV at a flow rate of 4 BV/h. The purity of total flavonoids increased to 61.95% after the purification, and the yield was 87.28%. Conclusion This optimized process was stable, feasible and suitable for separation and purification of total flavonoids from Tripterospermi Chinensis Herba.

Objective To study the adsorption technology of piceid on macroporous resin. Methods The extraction condition of piceid, the types of macroporous resin, the concentration of the sample of Rhizoma Polygoni cuspidati extract, the flow velocity of samples during adsorption were investigated and the leak curve was drawed to ascertain the optimal sample quantity. Results Resin D101 gave a better adsorption performance in the following technological conditions:the concentration of the sample of Rhizoma Polygoni cuspidati extract was 0.7774 mg/mL, the flow velocity of sample during adsorption 3BV/h, the optimal adsorption capacity of the resin was 10.88 mg/mL. Conclusion The procedure was simple, feasible and can be applied to industrial production.

Objective To evaluate the efficacy of surgical excision combined with iodine-125 implants intmcavitary radiotherapy for neumglioma Method After excision,27 patients with nenroglioma were implanted with the iodine-125 into the tumor bed or the residual tumor tissues.Results Of all cases,there were no operative mortality and no serious complication.The survival time of 25 cases exceeded 6 months and 1 glioblastoma recurrence,22 cases exceeded 12 months,3 glioblastoma and 1 astrocytoma Ⅲgrade recurrence.There were 19 cases exceeding 24 months,15 cases no recurrence,3 cases died for cere bral hernia because of giving.up further tremment,1 case surviving.Conclusions Surgical excision Combined with iodine-125 implants intracavitary radiotherapy can improve partial control rate,prolonged the survival time of patients with neuroglioma,and complications is rare.It is an efficient therapy for neu roglioma

Objective To explore the effect of Danshensu on myocardial mitochondrial function in diabetes cardiomyopathy(DCM)rats based on peroxisome proliferator-activated receptor γ(PPARγ)/peroxisome proliferator-activated receptor γ co-activator-1α(PGC-1α)pathway.Methods Rats were randomly separated into normal group,model group,low-(5 mg·kg-1),medium-(10 mg·kg-1)and high-(20 mg·kg-1)dose Danshensu groups,metformin group(140 mg·kg-1),as well as high-dose Danshensu+GW9662 group(20 mg·kg-1 Danshensu+1 mg·kg-1 GW9662),with 12 rats in each group.Except for the normal group,rats in other groups were fed with high-glucose and high-fat diet combined with intraperitoneal injection of streptozotocin to construct a DCM model.After successful modeling,the rats were administered corresponding drug once a day for six weeks.Fasting blood glucose values were detected by blood glucose meter.Echocardiography was applied to evaluate cardiac function of rats including left ventricular fractional shortening(LVFS)and left ventricular ejection fraction(LVEF).HE staining was applied to detect pathological changes in myocardial tissue.Transmission electron microscopy was applied to observe mitochondrial structure of myocardial tissue.JC-1 staining was applied to detect mitochondrial membrane potential in rat cardiomyocytes.The kit was applied to detect adenosine triphosphate(ATP)content and reactive oxygen species(ROS)expression in myocardial tissue.Western Blot was applied to detect the protein expression of PPARγ and PGC-1α in myocardial tissue.Results Compared with normal group,fasting blood glucose in model group was significantly increased(P＜0.05),LVFS and LVEF were significantly decreased(P＜0.05).It was found that myocardial tissue was obviously damaged and myocardial mitochondria became swollen.The percentage of non-deleted cardiomyocyte mitochondrial membrane potential was significantly decreased(P＜0.05).ATP content in myocardial tissue was significantly decreased(P＜0.05),ROS expression was significantly increased(P＜0.05).The protein expressions of PPARγ and PGC-1α in myocardial tissue were significantly downregulated(P＜0.05).Compared with model group,fasting blood glucose levels in Danshensu and metformin groups were significantly decreased(P＜0.05),while LVFS and LVEF were significantly increased(P＜0.05).It was found that myocardial tissue damage and mitochondrial structure damage were alleviated.The percentage of non-deleted cardiomyocyte mitochondrial membrane potential was significantly increased(P＜0.05).ATP content in myocardial tissue was significantly increased(P＜0.05),ROS expression was significantly decreased(P＜0.05).The protein expressions of PPARγ and PGC-1α in myocardial tissue were significantly upregulated(P＜0.05).Compared with high-dose Danshensu group,fasting blood glucose level in high-dose Danshensu+GW9662 group was significantly increased(P＜0.05),LVFS and LVEF levels were significantly decreased(P＜0.05).Damage of myocardial tissue and myocardial mitochondria structure became serious and myocardial mitochondria was obviously swollen.The percentage of non-deleted cardiomyocyte mitochondrial membrane potential was significantly decreased(P＜0.05).ATP content in myocardial tissue was significantly decreased(P＜0.05),ROS expression was significantly increased(P＜0.05).The protein expressions of PPARγ and PGC-1α in myocardial tissue were significantly downregulated(P＜0.05).Conclusion Danshensu improves mitochondrial function in DCM rats,which may be related to the activation of the PPARγ/PGC-1α pathway.

Objective To clone and express the heat shock cognate protein 20 (SjHsc20) of Schistosoma japonicum, and to preliminarily investigate its biological characteristics. Methods The target fragment of the SjHsc20 gene was amplified using PCR assay and cloned into the pET-28a(+) expression plasmid to generate the recombinant expression vector pET-28a(+)-SjH-sc20, which was then transformed into Escherichia coli BL21 (DE3) competent cells. The recombinant SjHsc20 (rSjHsc20) protein was induced with isopropyl β-D-thiogalactopyranoside (IPTG) and purified, and the expression of the rSjHsc20 protein was checked with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The immunogenicity of the rSjHsc20 protein was detected using Western blotting, and the transcriptional levels of SjHsc20 were quantified in S. japonicum worms at different developmental stages and in male and female adult worms using real-time quantitative PCR (RT-qPCR) assay. Thirty female BALB/c mice at ages 6 to 8 weeks were divided into three groups, including the rSjHsc20 immunization group, the PBS control group, and the ISA 206 adjuvant group, of 10 mice in each group. Mice in the rSjHsc20 immunization group were subcutaneously immunized with 20 μg rSjHsc20 on days 1, 15 and 31, and animals in the PBS control group were subcutaneously injected with the same volume of PBS on days 1, 15 and 31, while mice in the ISA 206 adjuvant group were subcutaneously immunized with the same volume of ISA 206 adjuvant on days 1, 15 and 31, respectively. All mice in each group were infected with (40 ± 2) S. japonicum cercariae via the abdomen 14 day following the last immunization. Levels of serum specific IgG and its subtypes IgG1 and IgG2 antibodies against rSjHsc20, and the serum titers of anti-rSjHsc20 antibody were detected in mice using indirect enzyme-linked immunosorbent assay (ELISA). All mice were sacrifice 42 days post-infection, and S. japonicum worms were collected from the hepatic portal vein and counted. The eggs per gram (EPG), worm burden reductions and egg burden reductions were estimated to evaluate the protective efficacy of the rSjHsc20 protein. Results The SjHsc20 gene had an open reading frame (ORF) with 756 bp in length and encoded 252 amino acids, and the rSjHsc20 protein had a relative molecular mass of approximately 29 kDa. The rSjHsc20 protein was recognized by the serum of mice infected with S. japonicum and the serum of mice immunized with the rSjHsc20 protein, indicating that rSjHsc20 had a good immunogenicity. There was a significant difference in the transcriptional levels of the SjHsc20 gene among the 7-day (1.001 4 ± 0.065 7), 12-day (2.268 3 ± 0.129 2), 21-day (1.378 5 ± 0.160 4), 28-day (1.196 4 ± 0.244 0), 35-day (1.646 3 ± 0.226 1), 42-day worms of S. japonicum (1.758 0 ± 0.611 1) (F = 38.45, P < 0.000 1), and the transcriptional level of the SjHsc20 gene was higher in the 12-day worms than in worms at other developmental stages (all P values < 0.000 1). The serum levels of anti-rSjHsc20 IgG antibody were 0.106 6 ± 0.010 7, 0.108 3 ± 0.010 4, and 0.553 2 ± 0.069 1 in the PBS control group, ISA 206 adjuvant group, and rSjHsc20 immunization group following the last immunization, respectively, and the serum levels of IgG1 antibody were 0.137 3 ± 0.054 0, 0.181 1 ± 0.096 8, and 1.765 8 ± 0.221 1, while the levels of IgG2a antibody were 0.280 3 ± 0.197 6, 0.274 0 ± 0.146 3, and 1.560 4 ± 0.106 0, respectively. There were significant differences in the serum levels of anti-rSjHsc20 IgG (F = 397.70, P < 0.000 1), IgG1 (F = 401.00, P < 0.000 1) and IgG2a antibodies (F = 229.70, P < 0.000 1) among the three groups, and the serum levels of anti-rSjHsc20 IgG, IgG1 and IgG2a antibodies were higher in the rSjHsc20 immunization group than in the PBS control group and the ISA 206 adjuvant group (all P values < 0.000 1). There was a significant difference in the IgG1/IgG2a ratio among the rSjHsc20 immunization group (1.177 2 ± 0.143 6), the PBS control group (0.428 4 ± 0.199 8) and the ISA 206 adjuvant group (0.559 9 ± 0.181 1) (F = 43.97, P < 0.000 1), and the IgG1/IgG2a ratio was > 1 in the rSjHsc20 immunization group, which was higher than in the PBS control group and the ISA 206 adjuvant group (both P values < 0.000 1). The titers of serum anti-rSjHsc20 antibody were all above 1∶16 384 in the rSjHsc20 immunization group following immunizations on days 1, 15 and 31, indicating that the rSjHsc20 protein had a strong immunogenicity. The mean worm burdens were (16.60±5.75), (15.80±5.58) worms per mouse and (14.40±5.75) worms per mouse in the PBS control group, the ISA 206 adjuvant group and the rSjHsc20 immunization group 42 days post-infection with S. japonicum cercariae (F = 0.50, P > 0.05), and the EPG were 68 370 ± 22 690, 67 972 ± 19 502, and 41 075 ± 13 251 in the PBS control group, the ISA 206 adjuvant group and the rSjHsc20 immunization group (F = 4.55, P < 0.05), with lower EPG in the PBS control group and the ISA 206 adjuvant group than in the rSjHsc20 immunization group (both P values < 0.05). Immunization with the rSjHsc20 protein resulted in a worm burden reduction of 13.25% and an egg burden reduction of 39.92% relative to the PBS control group. Conclusions SjHsc20 is successfully cloned and expressed, and the rSjHsc20 protein induces partial immunoprotective effects in mice, which provides a basis for deciphering the biological functions of SjHsc20 and assessing the potential of SjH-sc20 as a vaccine candidate.