AIM:To evaluate the bioequivalence of the two rivaroxaban tablets in Chinese healthy subjects.METHODS:Twenty-eight subjects under fasting status and twenty-eight subjects under fed status were enrolled in the study.This study was designed as a four period,fully repetitive,cross-over study.All subjects were administered test(T)and reference(R)rivaroxaban tablets(10 mg)un-der fasting and fed condition respectively.Liquid chromatography-tandem mass spectrometry was used to detect the concentrations of rivaroxaban in plasma.WinNonlin 7.0 was used to calculate the main pharmacokinetic parameters(PK)and to eval-uate the bioequivalence.RESULTS:In fasting group,the main pharmacokinetic parameters of T and R preparation were as follows:Cmax were(186.57±56.41)and(187.61±50.89)ng/mL;AUC0-t were(1 156.21±335.85)and(1 177.59±343.72)h·ng·mL-1;AUC0-∞ were(1 235.77±384.03)and(1223.53±392.10)ng·h·mL-1.The 90％confidential interval(CI)of the three main parameters were 90.81％-105.67％,92.83％-103.85％and 95.04％-107.13％.The upper limit of the 90％CI for the test-to-reference ratio of the within-subject of Cmax,AUC0-t and AUC0-∞ were 1.56,1.41 and 1.73.In fed group,the main pharmacokinetic parameters of T and R preparation were as follows:Cmax were(207.81±45.26)and(211.04±36.62)ng/mL;AUC0-t were(1 271.26±260.92)and(1 233.23±201.85)h·ng·mL-1;AUC0-∞ were(1 290.76±264.90)and(1251.68±203.73)ng·h·mL-1.The 90％CI of the three main parameters were 92.82％-102.28％,97.68％-106.68％and 97.71％-106.68％.The upper limit of the 90％CI for the test-to-reference ratio of the within-subject of Cmax,AUC0t and AUC0-∞were 1.76,1.47 and 1.47.CONCLUSION:The two preparations of rivaroxaban tablets were bioequiva-lent.

Objective:To investigate the genetic, clinical, and post-treatment characteristics of patients with glycogen storage disease type Ⅲ (GSD Ⅲ).Methods:A retrospective cohort analysis was performed on the genetic and clinical data of 26 cases with GSD Ⅲ who visited the Children's Hospital affiliated with Fudan University from June 2017 to December 2023. The patients were divided into non-missense variation and missense variation groups according to the types of mutation in the AGL gene.The correlation between genotype and phenotype was analyzed. All patients were treated with uncooked cornstarch after diagnosis. The changes before and after treatment were compared in patients who underwent more than twelve months of follow-up. A P value of <0.05 was used to denote statistical significance. Results:Among the 26 cases enrolled, 13 were female and 13 were male, and the median age of diagnosis was 28 (6 to 134) months. A total of thirty-five different types of AGL gene variation were detected, with c.1735+1G＞T (9/52, 17.3%) as the hotspot variation. The common clinical manifestations were elevated aminotransferases (26/26, 100%), hepatomegaly (25/26, 96.2%), fasting hypoglycemia (25/26, 96.2%), hyperketonemia (16/18, 88.9%), hypertriglyceridemia (TG) (20/26, 76.9%), elevated CK (16/25, 64.0%), and an abnormal electrocardiogram (12/16, 75.0%). Four cases (15.4%) had symptoms of myopathy at diagnosis. Liver biopsy was performed in eighteen cases, among whom 83.3% (15/18) had liver fibrosis≥S2. The number of cases with elevated levels of CK ( P＝0.031) and ALT ( P＝0.038)was pronounced in the non-missense variation group compared to that in the missense variation group. There were no statistically significant differences in age, height, liver size, degree of fibrosis, fasting blood glucose (Glu) and TG ( P>0.05). The median follow-up time of 14 cases was 40.5 (20-73) months, with improvement in body stature, reduced liver size, decreased ALT and TG, and improved Glu. However, four (28.6%) cases had new myopathy symptoms with raised CK ( P<0.05) and with advancing age, increased ALT diminished while CK level elevated ( P<0.05). Conclusions:The common clinical manifestations at the early stage of the GSD Ⅲdiagnosis are elevated aminotransferases, hepatomegaly, fasting hypoglycemia, hyperketonemia, high triglycerides, elevated CK, and fibrotic liver in China. Myopathy symptoms may arise following uncooked cornstarch treatment; however, there is significant improvement in height, liver-related, and metabolic parameters.

AIM:To evaluate the bioequivalence of the two rivaroxaban tablets in Chinese healthy subjects.METHODS:Twenty-eight subjects under fasting status and twenty-eight subjects under fed status were enrolled in the study.This study was designed as a four period,fully repetitive,cross-over study.All subjects were administered test(T)and reference(R)rivaroxaban tablets(10 mg)un-der fasting and fed condition respectively.Liquid chromatography-tandem mass spectrometry was used to detect the concentrations of rivaroxaban in plasma.WinNonlin 7.0 was used to calculate the main pharmacokinetic parameters(PK)and to eval-uate the bioequivalence.RESULTS:In fasting group,the main pharmacokinetic parameters of T and R preparation were as follows:Cmax were(186.57±56.41)and(187.61±50.89)ng/mL;AUC0-t were(1 156.21±335.85)and(1 177.59±343.72)h·ng·mL-1;AUC0-∞ were(1 235.77±384.03)and(1223.53±392.10)ng·h·mL-1.The 90％confidential interval(CI)of the three main parameters were 90.81％-105.67％,92.83％-103.85％and 95.04％-107.13％.The upper limit of the 90％CI for the test-to-reference ratio of the within-subject of Cmax,AUC0-t and AUC0-∞ were 1.56,1.41 and 1.73.In fed group,the main pharmacokinetic parameters of T and R preparation were as follows:Cmax were(207.81±45.26)and(211.04±36.62)ng/mL;AUC0-t were(1 271.26±260.92)and(1 233.23±201.85)h·ng·mL-1;AUC0-∞ were(1 290.76±264.90)and(1251.68±203.73)ng·h·mL-1.The 90％CI of the three main parameters were 92.82％-102.28％,97.68％-106.68％and 97.71％-106.68％.The upper limit of the 90％CI for the test-to-reference ratio of the within-subject of Cmax,AUC0t and AUC0-∞were 1.76,1.47 and 1.47.CONCLUSION:The two preparations of rivaroxaban tablets were bioequiva-lent.

OBJECTIVETo study the pharmacokinetics and bioequivalence of asparaginase loaded in hyaluronic acid-graft-poly(ethylene glycol)/ sulfobutylether-β-cyclodextrin nanocapsules (AHSP) in SD rats.

METHODSThe morphology of AHSP was observed under the transmission electron microscope and the particle size and zeta potential were measured. AHSP and free asparaginase were intravenously injected in rats, and the plasma asparaginase activity was measured at different time points after the injections. The pharmacokinetic parameters were calculated using the software DAS 2.1.1 to assess the bioequivalence of AHSP and free asparaginase.

RESULTSAHSP had an average particle size of 413.80∓10.97 nm with a zeta potential of -20.37∓2.38 mV. The AUC(0-48 h) of AHSP and free asparaginase was 137.34∓1.82 U/mL and 46.38 ∓1.98 U/mL, and their AUC(0-∞) was 164.66∓6.88 U/mL and 51.44∓3.01 U/mL with half-lives of 4.62∓0.60 h and 1.86∓0.38 h, respectively. Compared with free AN, AHSP exhibited increased AUC(0-48 h), AUC(0-∞), and half-life by 2.24, 2.55 and 2.32 folds, respectively. The 90% confidential intervals of AUC(0-48 h), AUC(0-∞) and Cmax of the tested formulation were 75.0%-76.5%, 74.3%-76.1%, and 95.1%-96.7%, respectively.

CONCLUSIONAHSP can improve the bioavailability and extend the biological half-life of asparaginase in rats, and AHSP and free asparaginase are not bioequivalent.

Animals ; Area Under Curve ; Asparaginase ; pharmacokinetics ; Biological Availability ; Half-Life ; Injections, Intravenous ; Nanocapsules ; Rats ; Rats, Sprague-Dawley ; Therapeutic Equivalency

Objective To screen specific human antibodies against Vibrio vulnificus hemolysin (VVH ) from natural single stranded large capacity phage antibody library and to perform initial identification of the antibodies,for the treatment of vibrio vulnificus infection.Methods The specific antibody against purified VVH was screened from large natural phage antibody library by 4 rounds of " adsorption-elution-amplification" screening process.Soluble ScFvs were prepared through infecting E coli.HB2151 and inducing expression with IPTG.The antigen binding activity and DNA sequence of the antibodies against VVH were identified by ELISA and DNA fingerprint analysis.Results After 4 rounds of panning,22 positive clones with specific binding ability to VVH were obtained,of which 8 obtained soluble expressions.DNA sequence analyses showed that the variable region genes of the clones belonged to different subgroups of immunoglobulin gene family.Conclusions The acquirement of human anti-VVH ScFv from large phage antibody library has laid a good foundation for the treatment of vibrio vulnificus infection.

Objective To screen specific human antibodies against Vibrio vulnificus hemolysin (VVH ) from natural single stranded large capacity phage antibody library and to perform initial identification of the antibodies,for the treatment of vibrio vulnificus infection.Methods The specific antibody against purified VVH was screened from large natural phage antibody library by 4 rounds of " adsorption-elution-amplification" screening process.Soluble ScFvs were prepared through infecting E coli.HB2151 and inducing expression with IPTG.The antigen binding activity and DNA sequence of the antibodies against VVH were identified by ELISA and DNA fingerprint analysis.Results After 4 rounds of panning,22 positive clones with specific binding ability to VVH were obtained,of which 8 obtained soluble expressions.DNA sequence analyses showed that the variable region genes of the clones belonged to different subgroups of immunoglobulin gene family.Conclusions The acquirement of human anti-VVH ScFv from large phage antibody library has laid a good foundation for the treatment of vibrio vulnificus infection.

Objective To construct recombinant expression vector TAT-KLF4 and express the Fusion Protein in E.coli BL21.Methods The open reading frame(ORF) of KLF4 gene was amplified by PCR.The PCR product was digested with restriction enzymes then was inserted into PET-28b-TAT-V2 which contained protein transduct domain of TAT.The recombinant vector was transfected into E.coli BL21 and the expression of the TAT-KLF4 fusion protein was induced with IPTG.The fusion protein was analyzed by using SDS-PAGE and purified by the affinity chromatography.Western Blot was used to identify the specificity of the fusion protein.The immunofluorescence was used to detection the efficiency of fusion protein transducted to HSF cells.Results The recombinant expression vector TAT-KLF4 was correctly constructed and fusion Proteion TAT-KLF4 was successfully expressed in prokaryotic cells.Western blotting showed the specificity of the fusion protein.The immunofluorescence prompt fusion protein transduced into HSF cells quickly.Conclusion This study provides the material basis for further induction of the induced pluripotent stem cells by protein transduction.

Objective To prepare the water soluble chrysin-hydroxypropyl-β-cyclodextrins inclusion compound and widen the administration path of chrysin .Methods The cogrinding method had been used to prepare chrysin-hydroxypropyl-β-cyclodextrins in-clusion compound .The PXRD, DSC and IR techniques had been used to characterize the inclusion compound .Results Chrysin and hydroxypropyl-β-cyclodextrins had formed the inclusion compound , and the formation of the inclusion compound could increase solubili-ty by 120.7 times.Conclusion The inclusion compound preparation method was simple and available , which was suitable to improve the bioavailability .

Objective To construct recombinant expression vector TAT-SOX2 and express the fusion protein in E.coli BL21.Methods SOX2 fragment were amplified by PCR.The product was digested with restriction enzymes then was inserted into PET-28b-TAT-V2 which contains protein transduction domain of TAT.The recombinant vector was transformed into E.coli BL and induced with IPTG.The expressed fusion protein was analyzed by using SDS-PAGE method.The highly expressed product was purified by affinity chromatography.Western blotting was used to identify the specificity of the fusion protein.The immunofluorescence was used to detect the efficiency of fusion protein transduced to HSF cells.Results The recombinant expression vector TAT-SOX2 was correctly constructed and fusion protein TAT-SOX2 was successfully expressed in prokaryotic cells.Western blotting showed the specificity of the fusion protein.The immunofluorescence prompt fusion protein was transduced into HSF cells quickly.Conclusion This study provides the material basis for the further induced pluripotent stem cells by protein transduction.

Objective To investigate the efficacy of CT guided radioactive 125I seed implantation on elderly patients with advanced malignancies.Methods 78 cases of elderly patients with malignant were collected and divided into two groups.41 cases of the treatment group were treated with CT guided radioactive 125I seed implantation.Particle activity was 29.6 MBq.The prescription dose was 90-110 Gy.37 cases of the control group were treated with optimized supportive care.Data from all of patients were to review and followup observation in short term efficacy,quality of life and side efforts.Results The total effective rate was 92.7 ％(38/41)and the disease control rate was 97.6 ％(40/41)in the treatment group.The control group was in the effective rate and 16.2 ％(16/37)in the disease control rate.The quality of life of the treatment group was higher than those in the control group(P＜0.05).And there is no obviously side efforts.Conclusion The treatment of elderly patients with advanced malignant tumors by 125I seed implantation was a safe and effective method.It can improve the quality of patients' life.