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A 13-year and 6-month-old girl attended the Hunan Children's Hospital due to delayed menarche. The laboratory test results indicated increased follicle-stimulating hormone and luteinizing hormone, decreased anti-Mullerian hormone, and pelvic ultrasound showed a cord-like uterus and absence of bilateral ovaries. Her 11-year and 5-month-old younger sister had the same laboratory and imaging findings, and both girls were diagnosed with primary ovarian insufficiency. Whole exome sequencing and Sanger sequencing confirmed that the proband and her sister carried heterozygous variants of HROB gene c.718C>T (p.Arg240*) and c.1351C>T (p.Arg451*), which were inherited from their parents respectively and consistent with autosomal recessive inheritance. Oral estradiol valerate at an initial dose of 0.125 mg/d was given to the proband, and the secondary sexual characteristics began to develop after 6 months.
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Humanos , Feminino , Criança , Lactente , Insuficiência Ovariana Primária/genética , Hormônio Luteinizante , EstradiolRESUMO
Objective:To observe the therapeutic effect of echinacoside (ECH) on liver injury and glucose metabolism disorder in sepsis rats induced by cecal ligation and puncture (CLP), and to explore its possible mechanism.Methods:Forty eight male Sprague Dawley (SD) rats were randomly divided into four groups: sham group (sham), model group (CLP), treatment group (CLP+ ECH) and inhibitor group (CLP+ ECH+ EX527). The sham group only received laparotomy, and the model group underwent CLP. The treatment group was intragastric administration of echinacea (30 mg/kg) every day after CLP modeling. The inhibitor group was injected with silence information regulator 1 (SIRT1) inhibitor EX527 (5 mg/kg) one hour before CLP, and then treated the same as the treatment group. Fasting blood glucose, insulin and serum biochemical indexes were detected in virous groups. The serum levels of interleukin (IL)-1 β, IL-6 and tumor necrosis factor-α(TNF-α) were detected by enzyme-linked immunosorbent assay (ELISA). 2′, 7′- dichlorofluorescein diacetate (DCFH-DA) staining was used to observe the production of reactive oxygen species (ROS) in liver tissue of rats in each group; hematoxylin-eosin (HE) staining was used to observe the pathological changes of liver tissue in each group; The expressions of SIRT1, glucose-6-phosphatase (G6Pase), phosphoenolpyruvate carboxykinase (PEPCK), phosphorylated signal transducers and activators of transcription 3 (p-STAT3) and phosphorylated protein ki-nase B(p-AKT) were detected by Western blot.Results:Compared with sham group, the levels of serum glucose, serum insulin, alanine aminotransferase (ALT), aspartate aminotransferase (AST), ROS, IL-1β, IL-6 and TNF-α in model group increased, while the liver glycogen and survival rate decreased (all P<0.05). After echinacoside treatment, the serum glucose, serum insulin, ALT, AST, ROS , IL-1β, IL-6 and TNF-α levels decreased, and the liver glycogen and survival rate increased (all P<0.05); After SIRT1 inhibitor intervention, the levels of serum insulin, ALT, AST, IL-6 and ROS in the inhibitor group increased ( P<0.05). HE staining showed that there were infiltration and necrosis of inflammatory cells in the liver tissue of model group, and echinacoside could significantly reduce the focal and massive necrosis; Western blot showed that compared with the sham group, the expression levels of SIRT1, p-STAT3 and p-AKT protein in the model group decreased, while the expression levels of G6Pase and PEPCK protein increased ( P<0.05); After echinacoside treatment, the expression levels of SIRT1, p-STAT3 and p-AKT increased, while the expression levels of G6Pase and PEPCK decreased ( P<0.05). After SIRT1 inhibitor intervention, the expression of SIRT1, p-STAT3 and p-AKT protein decreased, and the expression of G6Pase and PEPCK protein increased in the inhibitor group ( P<0.05). Conclusions:Echinacoside is a potential therapeutic agent for sepsis associated liver injury and glucose metabolism disorders, which may play a role by targeting SIRT1 to activate STAT3 and AKT in the liver.
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Objective:To improve the sensitivity and the linear range of electrochemical immunosensor to detect Schistosoma japonicum (S.japonicum) antibody.Methods:Carbon inks and silver/silver chloride inks were printed on a polyethylene terephthalate (PET) board to make a two-electrode test strip,where carbon was the working electrode and S.japonicum soluble egg antigen (SEA) was fixed at one end of working electrode by different methods; silver/silver chloride electrode was used as control.We tested the valency of the antibody by cyclic voltammetry (CV) and differential pulse voltammetry (DPV) in an electrochemistry workstation,and conducted comparison with the results of ELISA.Two new immunosensing electrodes have been developed,based on glutaraldehyde cross-linked (GA) or chitosan-glutaraldehyde cross-linked (Chit-GA) transducer fixing S.japonicum antigen.We tested the titer of the antibody by means of CV and DPV.Results:Our experimental S.japonicum antigen (50 μg/L) is the optimal test concentration for the GA sensor,and 10 μg/L for Chit-GA sensors.The immune reaction time of both electrodes is all essentially complete in 1 minute.The linear range for S.japonicura antibody in human positive serum sample detection by the glutaraldehyde cross-linked immunosensor is 1∶1000 to 1∶400,and by the chitosan-glutaraldehyde cross-linked immunosensor is 1∶1000 to 1∶500.As the concentration of dilution ratio of S.japonicum antibody in human positive serum sample increased,the test value of DPV increased proportionally.Conclusion:GA sensor and Chit-GA cross-linked S.japonicum sensors have high sensitivity and broad linear range response,and both exhibited a good linear relationship between the DPV signal and the test antibody titer.
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Objective To prepare the infected Oncomelania hupensis by artificial method for the research on the activity, vaccine, and genetic variation of Schistosoma Japonicum (S. Japonicum).Methods The mature eggs of S. Japonicum were collected by Nylon silk method and the miracidia were incubated under appropriate conditions. Negative snails were infected with miracidia in different proportion by means of individual or collective infection to seek the best method and proportion of infection between miracidia and snails. Infected snails were divided into 12 groups in total. Ⅰ-Ⅵ groups were for individual infection and Ⅶ-Ⅻ groups were for collective infection. There were 200 snails in each group. The infection ratios between snails and miracidia in Group Ⅰ-Ⅵ or screened, numbered, and reared singly. The amount of cercariae was calculated once every 10 days until the infected snails died. Then cercariae shedding quantity, infection quantity, and mortality of infected snails in every group were compared to find the best infection method and the best infection proportion between miracidia and snails. The cercariae were collected from the first generation of infected snails and were used to infect experimental animals. The mature eggs of S. Japonicum were saved from the infected experimental animals and incubated to get miracidia. The snails were artificially infected by miracidium to get the second generation of infected snails. The developmental rates of adult worms, the egg density in fecal and liver were compared between artificially and naturally infected snails. Results In individual infection GroupⅠ-Ⅵ,the average infection value of snails were 0±0,22.7±4.2,31.7±4.5,53.0±5.3,39.3±5.9,32.7±4.7,the average fatality of snails were 21.7±3.1,25.0±3.6,31.3±4.9,44.7±6.5,78.3±9.5,89.7±13.6, and the average value of cercariae shedding from infected snails were 0.0±0.0,308.0±96.6,428.1±146.2,527.0±171.1,571.4±148.9,602.9±356.3, respectively. In collective infection Group Ⅶ-Ⅻ,the average infection value of snails were 0±0,12.3±2.5,18.7±4.7,28.3±4.2,33.3±4.7,29.3±5.5,and the average fatality of snails were 22.7±3.8,23.7±4.5,28.3±5.5,47.0±9.5,75.7±8.5,86.3±12.2, and the average value of cercariae shedding from infected snails were 0±0,244.5±57.3,292.3±74.8,347.1±100.8,477.2±142.1,447.3±161.4, respectively. The second generation of artificially infected snails was obtained successfully. The average infection rate and fatality rate for the second generation of artificially infected snails were 24.65% and 24.50%, both of which were not obviously different from that of the first generation of artificially infected snails (P>0.05). In the animal experiment, the worm growth rate for the naturally infected snails, the first or second generation of artificially infected snails were 68.50%,73.50% or 71.00%. There was no obvious difference among them (P>0.05). The fecal (or liver) eggs per gram for the naturally infected snails, the first or the second generation of artificially infected snails were 1 503±269,1 683±233, or 1 541±117 (or 6 641±1 819,6 272±1 419, or 7 263±1 643). There was no significant difference among the 3 groups (P>0.05). Conclusion Infected snails can be obtained through the artificial method by using S. Japonicum miracidia to infect snails. Individual infection has the advantage over collective infection. The optimal proportion of infection between first and the second generation of artificially infected snails in the average of cercariae shedding, infection, and fatality average of snails. There was no significant difference between artificially and naturally infected snails in the developmental rate of adult worms, fecal and liver eggs per gram.