OBJECTIVETo develop an assay methodology for determination of lumbrukinase potency in Rongshuan capsules.

METHODThe agarose-fibrin plate assay methodology for determination of Lumbrukinase potency in Rongshuan capsules was studied including durability, specificity, linearity range, product's handling method, accuracy , repetitiveness, solution stability, recovery and statistical method. The method of parallel line assay based on quantitative responses in statistical methods for biological assays was used in the statistics of potency assay.

RESULTThe durability and specificity of assay accord with the requirement; The linearity range was 12.5 to approximately 400 U, the RSD of accuracy tests was 3.2%, the RSD of repetitiveness tests was 8.3%, the solution is stable under 4 degrees C for 72 hours, the recovery rate was 97.0% and the RSD of recovery assays was 16.5%.

CONCLUSIONThe agar-fibrin plate assay is rapidly, feasible, simple, convenient and accurate way for determining the Lumbrukinase potency. The method of parallel line assay based on quantitative responses in statistical methods for biological assays can control the error of determination.

Animals ; Biological Assay ; methods ; Capsules ; analysis ; Endopeptidases ; analysis ; Pharmaceutical Preparations ; analysis

Objective:To investigate the killing effect of radiation on echinococcus in vitro culture and its effect on the mRNA expression of growth arrest and DNA damage 45 alpha (Gadd45α) gene. Methods:Echinococcus from naturally infected sheep liver was cultured in vitro and divided into 7 groups. The echinococcus was irradiated with 6 MeV at doses of 0 (control), 20, 40, 60, 80, 100, and 120 Gy, respectively. The growth of echinococcus was observed under light microscope at 1, 3, 5 and 7 d after the radiation. The expression of Gadd45α mRNA in control, 20, 40 and 60 Gy groups of echinococcus was detected by RT-PCR technique at 7 d after the radiation. Results:The disintegration and exfoliation of echinococcus were observed under light microscope at 1, 3, 5 and 7 d after the radiation, and the death rate of echinococcus was positively correlated with the radiation dosages ( r = 0.81, P < 0.05). After the radiation at 7 d, compared with the control group (100.00 ± 0.00), the mRNA expression levels of Gadd45α in echinococcus of 20, 40, and 60 Gy groups were significantly increased (279.74 ± 80.08, 759.38 ± 160.98, 1 666.68 ± 316.36, P < 0.01), and it was positively correlated with the radiation dosages ( r = 0.93, P < 0.01). Conclusion:Radiation has a certain killing effect on echinococcus cultured in vitro, and there is a certain dose-effect relationship with the radiation dosages, and Gadd45α gene may be involved in the molecular mechanism of radiation-induced killing of echinococcus in vitro.

OBJECTIVE: To discuss the value of chromosomal microarray analysis (CMA) for the identification of DMD gene deletions during prenatal diagnosis. METHODS: G-banded karyotyping and CMA were performed on fetuses with ultrasonographic soft markers but no family history for Duchenne/Becker muscular dystrophy (DMD/BMD). Denaturing high-performance liquid chromatograghy (DHPLC) was used to detect DMD gene mutations in umbilical cord blood and peripheral blood samples from the mothers. RESULTS: For fetus 1, analysis of amniocytes showed a normal karyotype, while CMA detected a 119 kb deletion at Xp21.1 (32 565 489 - 32 681 461), which encompassed exons 10 to 16 of the DMD gene. The result was confirmed by DHPLC analysis. The mother was found to have loss of heterozygosity in the same region. For fetus 2, karyotyping of amniocytes also showed a normal male karyotype, while CMA detected a 254 kb deletion at Xp21.1 (32 104 604 - 32 358 874), which encompassed exons 41 to 44 of the DMD gene. The same deletion was not detected in the mother. DHPLC analysis confirmed the presence of both deletions. CONCLUSION Two fetuses harboring DMD gene deletions but without a family history were discovered. CMA can improve the efficiency for detecting single gene diseases caused by deletions.
Dystrophin ; genetics ; Exons ; Female ; Fetus ; Gene Deletion ; Humans ; Incidental Findings ; Male ; Microarray Analysis ; Muscular Dystrophy, Duchenne ; genetics ; Pregnancy

We hereby reported a case of ring chromosome 18 complicated by the deletion of 18p11.32p11.31 and 18q21.33q23 diagnosed prenatally by G-banding karyotype and chromosomal microarray analysis (CMA). Ultrasound scan indicated a single umbilical artery and intrauterine growth retardation at the second trimester. The result of G-banding karyotyping was 46, XN, r(18)(p11.3q21.3) and CMA indicated that there was a 3.3 Mb deletion at 18p11.32p11.31 and a 16.9 Mb deletion at 18q21.33q23. All these suggested that the fetus might present with clinical manifestations such as growth retardation, epilepsy, speech delay and growth hormone deficiency after birth, so the couple decided to terminate the pregnancy after genetic counseling.

Objective: To investigate the prenatal ultrasonographic features and prognosis of vasa previa, to explore the application value of sector scanning in the intracervical mouth by antenatal ultrasound, then to increase vasa previa detection rate. Methods: Prenatal ultrasound images, clinical characteristics and pregnancy outcome of 35 pregnant women with vasa previa confirmed by surgery and pathology were analyzed retrospectively, the diagnostic effectiveness of sector scanning in the intracervical mouth was evaluated. Results: Thirty-three of the 35 vasa previa cases were detected by sector scanning in the intracervical mouth, with a detection rate of 94.3% (33/35). Of the 35 cases, 20 cases (60.6%) were first contacted in second trimester and 13 cases (39.4%) were first contacted in third trimester. Two cases were missed or misdiagnosed, which were all first contact in third trimester. Among the 35 cases, 25 were velamentous placenta and 4 were battledore placenta. Twenty cases were low-lying placenta or marginal placenta previa. All 35 women underwent cesarean section. No neonatal mortality, 11 term infants, 20 premature infants of more than 34 weeks and 4 premature infants of less than 34 weeks. All placentas underwent pathological examination after delivery, 4 cases placentas underwent vascular casting, and it was found that 2 cases were vasa previa of umbilical artery branch and 2 cases were vasa previa of allantoic veins branch. Conclusions Vasa previa can be effectively detected by prenatal ultrasonography through sector scanning in the intracervical mouth. Second trimester is the best period to detect vasa previa. Pathomorphological examination on placenta after delivery and vascular casting are helpful to the understanding of vasa previa.

Glycolytic metabolism enzymes have been implicated in the immunometabolism field through changes in metabolic status. PGK1 is a catalytic enzyme in the glycolytic pathway. Here, we set up a high-throughput screen platform to identify PGK1 inhibitors. DC-PGKI is an ATP-competitive inhibitor of PGK1 with an affinity of K _d = 99.08 nmol/L. DC-PGKI stabilizes PGK1 in vitro and in vivo, and suppresses both glycolytic activity and the kinase function of PGK1. In addition, DC-PGKI unveils that PGK1 regulates production of IL-1β and IL-6 in LPS-stimulated macrophages. Mechanistically, inhibition of PGK1 with DC-PGKI results in NRF2 (nuclear factor-erythroid factor 2-related factor 2, NFE2L2) accumulation, then NRF2 translocates to the nucleus and binds to the proximity region of Il-1β and Il-6 genes, and inhibits LPS-induced expression of these genes. DC-PGKI ameliorates colitis in the dextran sulfate sodium (DSS)-induced colitis mouse model. These data support PGK1 as a regulator of macrophages and suggest potential utility of PGK1 inhibitors in the treatment of inflammatory bowel disease.