Objective:To explore the association between the CYP21A2 genotype and the virilization severity in girls with classic 21-hydroxylase deficiency (21-OHD), so as to further the understanding of virilization in females and provide guidance for prenatal diagnosis and genetic counseling. Methods:A total of 23 patients with two X chromosomes (46, XX) who were newly diagnosed with classic 21-OHD in Wuhan Children′s Hospital from August 2010 to March 2019 were included.These patients were divided into 3 groups according to the Prader grades of the degree of external genitalia masculinization.The 17-hydroxyprogesterone (17-OHP) level, androstenedione (AND) level, testosterone (T) level, dehydroepiandrosterone sulphate (DHEAS) level and genotypes were recorded.The gene mutations were divided into the Null group(the enzyme activity was completely impaired), group A(1% of the normal enzyme activity was retained), group B(2% of the normal enzyme activity was retained) and group C(20%-60% of the normal enzyme activity was retained). The correlation between the gene variation of different Prader grades and the corresponding gene groups were analyzed.Results:All 23 girls showed different degrees of external genitalia masculi-nization.There was a significant positive correlation between the Prader grades and the type of gene variation ( rs=0.696, P<0.001). The gene group A and Null group were highly matched with the Prader Ⅳ(the matching rate: 77.8%). Eighty percent of girls with Prader Ⅱ and Prader Ⅲ were classified as the gene group B. The testosterone le-vel of girls with Prader Ⅳ at first diagnosis was significantly higher than that of girls with Prader Ⅱ [4.6(4.0, 15.0)μg/L vs.0.63(0.40, 1.39)μg/L]( χ2=15.117, P<0.05). Conclusions:There is a significantly positive and strong correlation between the degree of external genitalia masculinization and the degree of deficiency of enzyme activity caused by gene variation in girls with typical 21-OHD.It can provide reference for both parents carrying CYP21A2 gene in prenatal diagnosis, genetic counseling and individualized diagnosis and treatment.

Objective To compare the efficacy and efficiency of simulation-based training of flexible fibreoptic intubation in novices with virtual reality simulator.Methods A total of 46 anaesthesia residents in their first stage of training in anaesthesiology with no experience in flexible fibreoptic intubation at Peking University People's Hospital were enrolled in the study,and were divided into 2 groups randomly,which were virtual reality simulator group (group S,n=23) and manikin group (group M,n=23).The group S was then trained for 25 times on simulator,while the group M did the same processes on manikin.After training,participants in both groups had their performance assessed with the fibrescope evaluated through the oral route using a simulation manikin,who were instructed to attempt to advance the fibrescope 5 consecutive times to view the carina in the shortest amount of time.The time required to view the carina of each practice during training in both groups were recorded as pooled data to construct group learning curves with the application of SPSS 20.0.By using repeated measures analysis of variance and Ttest,the procedure time and global rating scale (GRS) of fibreoptic bronchoscope manipulation ability were compared between groups,so did the participant's confidence between before and after the training both within-subjects and between-subjects.Results The plateaus in the learning curves were achieved after 19 (15,26) practice sessions in group S and 24 (19,31) in group M,respectively.There was no significant difference in the procedure time [(13.7 ± 6.6) s and (11.9 ±4.1) s] and GRS [(3.9 ±0.4) vs.(3.7 ±0.3)]between groups.There were significant increases in participant's confidences in both groups after training [group S:(1.8 ± 0.5) vs.(3.9 ± 0.6),t=10.928,P=0.000;group M:(2.0 ± 0.7) vs.(3.9 ± 0.5),t=15.306,P=0.000],but there was no significant difference between groups.Conclusion The simulation-based training of flexible fibreoptic intubation in novices with virtual reality simulator is more efficient than the one with manikin,but the similar effects can be achieved in both modalities,after adequate trainings.In the related training a balance between time cost and economic cost should be considered and the appropriate teaching methods and forms should be taken.

Objective To evaluate the precision of BREEZE~(TM) glucose monitor and correlation of glucose measurements between fingertip capillary whole blood glucose(CBG)using BREEZE~(TM) glucose monitor and venous plasma glucose(VPG)using autoanalyzer.Methods All samples of venous plasma and fingertip blood from 188 diabetes or non-diabetes patients were detected for glucose level at fasting,30,60 and 120 min postprandial.BREEZE~(TM) glucose monitor and autoanalyzer measured CBG and VPG respectively.Intra-and inter- coefficients of variations were determined using 10 BREEZE~(TM) glucose monitors with normal,slight high and high glucose levels for the three lots of strips.Results The correlation coefficients between CBG and VPG were all higher than 0.950 at fasting and different postprandial time.98.94% of all measurements were in the A zone when using error-grid analysis.The relevant differences between CBG and VPG were less than 5% at different blood glucose concentrations.The intra-and inter-coefficients of variations of blood glucose values at different blood glucose concentrations using different lots of strips were within 5%.Conclusion BREEZE~(TM) glucose monitor provides high accurate and precise glucose readings on fasting and different postprandial time points over a variety of blood glucose concentrations.

Aim To observe the effects of the ethyl ace-tate extract from Coreopsis tinctoria on glucose and lipid metabolism and liver, kidney function in diabetic rats. Method By high-sugar, high-fat diet combined with intraperitoneal injection of streptozotocin ( streptozoto-cin, STZ ) Type 2 diabetes SD rat model was estab-lished. Model rats were randomly divided into six groups ( control group, model group, three dose groups Coreopsis tinctoria extract:low, middle,high 0. 15 g· kg-1;0. 3 g·kg-1;0. 6 g·kg-1 , positive drug met-formin 0. 16 g · kg-1 group ) . The control group and the model group were given physiological saline and the remaining groups intragastric administration coreofosis tinctoria extrat. Random blood glucose and body weight of rats were measured weekly. After 4 weeks of admin-istration, The rats were killed and rat serum was col-lected to detect serum lipids ( TC/TG/HDL/LDL ) , liver and renal function indicators, serum insulin, and glycated hemoglobin levels. Result Coreopsis tincto-ria ethyl acetate extract effectively reduced the diabetic rats random blood glucose, glycated hemoglobin,serum triglycerides, LDL, total serum protein, serum creati-nine and uric acid levels, and increased serum white protein content in diabetic rats. Conclusion Coreop-sis tinctoria ethyl acetate extract can reduce blood glu-cose and lipid in diabetic SD rats and protect their liver and kidney function.

Adult ; Fabry Disease ; pathology ; Female ; Follow-Up Studies ; Humans ; Kidney Glomerulus ; pathology ; ultrastructure ; Proteinuria

In order to identify Aucklandiae Radix, Vladimiriae Radix, Inulae Radix, Aristolochiae Radix and Kadsurae Radix using ITS2 barcodes, genomic DNA from sixty samples was extracted and the ITS2 (internal transcribed spacer) regions were amplified and sequenced. The genetic distances were computed using MEGA 5.0 in accordance with the kimura 2-parameter (K2P) model and the neighbor-joining (NJ) phylogenetic tree was constructed. The results indicated that for Aucklandiae Radix (Aucklandia lappa), Vladimiriae Radix (Vladimiria souliei and V. souliei var. cinerea), Inulae Radix (Inula helenium), Aristolochiae Radix (Aristolochia debilis) and Kadsurae Radix (Kadsura longipedunculata), the intra-specific variation was smaller than inter-specific one. There are 162 variable sites among 272 bp after alignment of all ITS2 sequence haplotypes. For each species, the intra-specific genetic distances were also smaller than inter-specific one. Furthermore, the NJ tree strongly supported that Aucklandiae Radix, Vladimiriae Radix, Inulae Radix, Aristolochiae Radix and Kadsurae Radix can be differentiated. At the same time, V. souliei (Dolomiaea souliei) and V. souliei var. cinerea( D. souliei var. cinerea) belonging to Vladimiriae Radix were clearly identified. In conclusion, ITS2 barcode could be used to identify Aucklandiae Radix, Vladimiriae Radix, Inulae Radix, Aristolochiae Radix and Kadsurae Radix. Our study may provide a scientific foundation for clinical safe use of the traditional Chinese medicines.
Aristolochia ; classification ; genetics ; Base Sequence ; DNA Barcoding, Taxonomic ; methods ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Drugs, Chinese Herbal ; chemistry ; classification ; Molecular Sequence Data ; Phylogeny ; Plants, Medicinal ; classification ; genetics ; Quality Control

OBJECTIVEBy using the RNAi method to inhibit Itk protein expression specificity, to observe lymphocytes proliferation and cytokines production, verify its function as a drug target.

METHODSDesigned siRNA aims at Itk sequence according to its sequence and solid structure, then electrotransfected into mouse spleen lymphocytes, We validated the decrease of Itk protein by Western-Blot, and detected the change of the cell proliferation by MTS and the change of inflammatory cytokines by ELISA.

RESULTSItk protein can be suppressed by Itk-siRNA, there were significantly reduced compared to its control group on cell proliferation as well as cytokine secretion such as IL-2, IL-4, IL-5, IFN-gamma. They all have statistical difference (P < 0.05).

CONCLUSIONItk has an important immunomodulatory effect in mouse spleen lymphocytes proliferation and secretion of inflammatory cytokines.This can supply an experimental basis to regard Itk as drug target for inflammation therapy.

Animals ; Cell Differentiation ; Cell Proliferation ; Cytokines ; genetics ; immunology ; Lymphocytes ; cytology ; immunology ; Male ; Mice ; Mice, Inbred BALB C ; Protein-Tyrosine Kinases ; genetics ; immunology ; Spleen ; cytology ; immunology

Several kinds of column chromatography method were used to investigate the chemical constituents of the leaves of Lophatherum gracile. The structures of the isolated compounds were identified based on their physicochemical properties and spectral data. A new flavone C-glycoside was isolated and its structure was identified as 3'-methoxyl-luteolin 6-C-beta-D-galactopyranosiduronic acid (1 --> 2) -alpha-L-arabinopyranoside (1).
Antiviral Agents ; chemistry ; isolation & purification ; Flavones ; chemistry ; Glycosides ; chemistry ; isolation & purification ; Hydrolysis ; Plant Leaves ; chemistry ; Poaceae ; chemistry

Site-directed mutagenesis method was used to introduce two desired mutations, which were confirmed by DNA sequencing, into mouse complement receptor Type II gene(MCR2). Then the constructed eukaryotic expression vectors containing wild type mouse CR2/1(wtMCR2/1), mutant type mouse CR2/1 (mtMCR2/1) and human CR2 (hCR2) cDNA were transferred into mouse SP2/0 cells by electroporation. After two-week screening by G418, the stably transfected clones were obtained. Several ways including PCR, RT-PCR, and immunohistochemistry were utilized to screen those clones with interesting genes integrated and expressed. Then Epstein-Barr virus(EBV) was used to infect these transfected cells and EBER-1 (EBV encoded RNAs) hybridization results showed that only hCR2 and mtMCR2 transfected SP2/0 cells could be infected by EBV, but positive rate of the former was much higher than the latter. This study sets groundwork for elucidating the mechanism by which EBV enters the cells and for establishing the animal model of EBV-related nasopharyngeal carcinoma (NPC).

Previously the diseases of pediatric hyperandrogenism were mainly diagnosed and evaluated by testing traditional androgens such as testosterone and androstenedione.However, clinical application has revealed a poor correlation between traditional androgens and the clinical manifestations of hyperandrogenism in some patients.It has been proposed that adrenal-derived 11-oxygenated androgen may also be involved in the course of this type of disease.The concentrations of 11-oxygenated androgen are elevated in androgen excess diseases, and they fulfill a variety of roles in human physiology and disease.This article discusses three aspects of the synthesis process, activity and content of 11-oxygenated androgen and their application in three androgen excess diseases: congenital adrenocortical hyperplasia, premature adrenarche and polycystic ovary syndrome, in order to help clinicians expand their clinical understanding and investigative thoughts on 11-oxygenated androgen.