【Objective】 To investigate the possible molecular pathogenesis of a child with hemophilia A accompanied by coagulation factor Ⅺ reduction by testing coagulation-related indicators and genotyping in the child and his family. 【Methods】 Peripheral blood from the patient and his parents for detection of coagulation factors Ⅷ, Ⅸ, Ⅺ, Ⅻ, VWF∶Ag, lupus anticoagulants and F VIII, F XI inhibitors were collected. All exons and flanking sequences of the genes encoding FⅧ and FⅪ were sequenced and bioinformatically analyzed. 【Results】 The child had low FⅧ and FⅪ activity and no parental abnormalities were observed. The sequencing results showed that there was a c. 1834(exon12) C>T heterozygous mutation in the FⅧ gene and a c. 1817 (exon15) G>A heterozygous mutation in the FⅪ gene, which was de novo. Bioinformatics analysis shows that the FⅪ mutation changes the original protein structure and increases the number of carboxyl groups. 【Conclusion】 For patients with prolonged APTT, in addition to excluding factors that interfere with APTT testing, all coagulation factors related to APTT should be tested to clarify the diagnosis.

Humans ; Infant ; Biliary Atresia/diagnosis* ; Choledochal Cyst/diagnosis* ; RNA, Circular ; Diagnosis, Differential

Objective To investigate the role of nitric oxide synthase(NOS)in the regulation of myocardial ischemia-reperfusion(IR)injury in ovariectomized(OVX)rats.Methods A total of 132 female SD rats were subjected,and 48 of them were randomly divided into sham operation group,IR group,OVX group and combined group,with 12 in each group.In order to explore the role of endothelous NOS(eNOS)and inducible NOS(iNOS)in ovariectomization increasing myo-cardial IR injury,another 84 mice were divided into negative sham group,negative IR group,nega-tive combined group,eNOS+IR group,eNOS combined group,iNOS small interfering RNA(si-iNOS)+IR group and si-iNOS combined group,with 12 in each group.The mice of the corre-sponding groups were injected with adeno-associated virus(AAV)overexpressing eNOS or knoc-king down iNOS via tail vein before OVX modeling.Myocardial infarct size,serum levels of lac-tate dehydrogenase(LDH)and creatine phosphokinase isoenzyme(CK-MB),LVEF,LVFS,and expression levels of eNOS and iNOS in the myocardial tissues were measured.Results The com-bined group had significantly increased level of iNOS in myocardium,larger myocardial infarct size and elevated serum LDH and CK-MB levels,but decreased myocardial expression of eNOS and LVEF and LVFS values than the IR group(P＜0.05).When compared with the negative combined group,the myocardial infarct size and serum LDH and CK-MB levels were decreased[(23.51±3.22)％and(26.21±2.93)％vs(58.78±5.42)％,(176.31±15.48 and 169.52±17.12 vs 328.85±37.12 U/L,35.41±6.41 and 34.77±5.94 vs 88.73±9.14 U/L,P＜0.05],and the LVEF and LVFS values were increased[(41.31±3.12)％and(42.09±3.41)％vs(30.77± 2.15)％,(21.47±1.57)％and(21.32±1.42)％vs(15.92±1.33)％,P＜0.05]in the eNOS com-bined group and si-iNOS combined group.Conclusion The decrease of eNOS expression and in-crease of iNOS expression are related to the aggravation of myocardial IR injury in OVX rats.

Objective:To explore the effect of auricular point pressing pill combined with rhubarb acupoint application on intestinal preparation and prevention of adverse events in colonoscopy.Methods:A total of 220 patients who met the inclusion criteria from May 2019 to February 2020 were randomly divided into 4 groups, 55 in each group. The control group was only given compound polyethylene glycol electrolyte powder. The auricular plaster group was combined with auricular point pressing pill on the basis of the control group. The acupoint sticking group was combined with rhubarb acupoint application on the basis of the control group. The combination group was combined with auricular point pressing pill and rhubarb acupoint application on the basis of the control group. The quality of bowel preparation was evaluated by Boston Bowel Preparation Scale(BBPS), and the adverse events such as perianal discomfort, defecation discomfort, abdominal pain and abdominal distension were observed.Results:After treatment, there was significant difference in BBPS score among the four groups ( F=3.038, P=0.030). The BBPS score in the acupoint sticking group (7.45 ± 1.20 vs. 6.78 ± 1.32) and combination group (7.26 ± 1.11 vs. 6.78 ± 1.32) were significantly higher than those in the control group( P<0.05). After treatment, there were significant differences in abdominal pain and abdominal distension scores among the four groups ( F=2.947, 5.879, P=0.034, 0.001). The scores of perianal discomfort (0.07 ± 0.38 vs. 0.44 ± 1.07), defecation discomfort (0.40 ± 0.81 vs. 0.87 ± 1.32) and abdominal pain (0.93 ± 1.32 vs. 1.69 ± 1.61) in the combination group were significantly lower than those in the control group ( P<0.05), and the abdominal distension score (1.05 ± 1.00 vs. 1.64 ± 0.95) in the acupoint sticking was significantly lower than that of the control group ( P<0.05). Conclusion:Auricular combined with rhubarb powder acupoint application can improve the quality of bowel preparation and reduce intestinal adverse reactions. It can be used with polyethylene glycol electrolyte powder for the intestinal preparation process.

Thyroid cancer is the most common head and neck malignant tumor, and its incidence is increasing year by year. Therefore, it is of great significance to explore the mechanism of the occurrence and development of thyroid cancer, and develop the diagnostic markers and therapeutic targets of thyroid cancer, so as to improve the curative effect of thyroid cancer and prolong the survival time of patients. The previous study has found that long non-coding RNA (lncRNA) DANCR has abnormal high expression in thyroid cancer, which could promote the expression of leucine-rich repeat kinase 2 (LRRK2) by affecting miRNA-185-5p and transcription factor SMARCB1, and indirectly participate in autophagy and regulate the development of thyroid cancer. LncRNA DANCR is expected to become a new target for diagnosis and treatment of thyroid cancer. This paper introduces the possible regulatory mechanism of lncRNA DANCR in thyroid cancer, and provides new ideas for the basic research and clinical diagnosis and treatment of thyroid cancer.

Objective:To investigate the effect of edaravone, a free radical scavenger, on the regulation of retinal autophagy and the protection of photoreceptor cells at the early stage of experimental retinal detachment (RD) in rats.Methods:Fifty-one adult male Sprague-Dawley rats were used for RD model establishment, and another 24 rats were served as PBS injection group.The RD model was established via subretinal injection of 0.5% sodium hyaluronate into the right eye of the rats and the rats with successful modeling were randomly divided into RD model group and edaravone treatment group.The rats in the edaravone treatment group were given edaravone of 3 mg/kg intraperitoneally, twice a day after modeling, and the rats in the PBS injection group and RD model group were given equal volume of normal saline.Rats were sacrificed on the 1st day, 3rd day and 7th day following modeling.The T-superoxide dismutase (T-SOD) activity and malondialdehyde (MDA) content in the intraocular fluid was detected.The expression levels of superoxide dismutase 2 (SOD2), nuclear factor E2-related factor 2 (Nrf2), autophagy related gene 4 (Atg4), microtubule-associated protein 1 light chain 3B (LC3B) and other proteins in retinal tissue were identified by Western blot analysis.TUNEL staining was performed on paraffin sections of the whole eyeball to analyze the apoptosis of photoreceptor cells.The study protocol was approved by an Ethics Committee of Xi'an Fourth Hospital (No. 2016016). The use and care of animals complied with the Regulations on the Administration of Experimental Animals.Results:The RD area was more than 60% in rat eyes of RD model.There were significant differences in MDA content and T-SOD activity among different groups at various time points (MDA: Fgroup=385.513, P<0.01; Ftime=13.021, P<0.01.T-SOD: Fgroup=48.865, P<0.01; Ftime=7.700, P=0.003). Compared with the PBS injection group, the MDA concentration was significantly increased and the T-SOD activity was significantly decreased in the RD group and edaravone treatment group on the 1st, 3rd and 7th day after modeling (all at P<0.05). The MDA concentration was significantly reduced and the T-SOD activity was significantly elevated in the edaravone treatment group on the 1st, 3rd and 7th day after modeling in comparison with those of the RD group (all at P<0.05). Compared with the PBS injection group, the relative expression levels of SOD2 and Nrf2 proteins were significantly increased in the RD group and edaravone treatment group on the 1st, 3rd and 7th day after modeling (all at P<0.05), and Atg4 and LC3B-Ⅱ/LC3B-Ⅰ were significantly increased on the 1st, 3rd and 7th day after modeling (all at P<0.05). The expression level of SOD2 in the edaravone treatment group was significantly higher than that in the RD group on the 1st, 3rd and 7th day after modeling (all at P<0.05), and the expression level of Nrf2 was significantly increased in the edaravone treatment group on the 1st and 3rd day after modeling compared with that of the RD group (both at P<0.05), and the expression levels of Atg4 and LC3B-Ⅱ/LC3B-Ⅰ were significantly increased in the edaravone treatment group on the 3rd day after modeling in comparison with those of the RD group (both at P<0.05). No significant TUNEL positive cells were observed in PBS injection group at all time points, and TUNEL positive cells were observed on the 1st, 3rd and 7th day after modeling in the RD group, and the expression level of caspase-3 in the RD group was significantly increased in comparison with that of the PBS injection group ( P<0.05). The apoptosis of photoreceptor cells and the expression level of caspase-3 in edaravone treatment group were significantly decreased in comparison with those of the RD group on the 1st, 3rd and 7th day after modeling (all at P<0.05). Conclusions:The intraperitoneal injection of edaravone, twice a day, can significantly improve the antioxidant capacity of the retina after experimental RD in rats, regulate retinal autophagy and reduce the apoptosis of photoreceptor cells in early-stage RD.

Objective: To determine the expression of TAZ and its role in angiogenesis in gastric carcinoma. Methods: Immunohistochemical staining was performed to investigate the expression of TAZ and to determine whether a direct relationship exists between TAZ and β-catenin. Transfection with TAZ overexpression plasmid in MKN28 cells was conducted to induce exogenous expression of TAZ and a TAZ knockdown plasmid was transfected into MGC803 cells to reduce TAZ levels. The effects on endothelial cell formation, proliferation, and migration were determined by Matrigel three-dimensional culture, MTT proliferation assay and Transwell migration assay. In addition, the expression of TAZ and β-catenin in transfected gastric cancer cells was detected by Western blot. Results: Immunohistochemistry showed that TAZ protein was expressed in 64 of 150 gastric cancer sample tissues (43%), TAZ was localized in the nucleus, and its expression was associated with tumor grade, TNM stage, metastasis, and microvessel density (MVD) (P<0.05). In addition, the expression frequency of β-catenin in the TAZ positive group was 67.2%, which was significantly higher than that in the TAZ negative group, and the expression of TAZ was positively correlated with β-catenin. After transfection, TAZ overexpression increased the expression of β-catenin and enhanced HUVECs tube formation, proliferation, and migration. In the MGC803 cells transfected with the knockdown plasmid, β-catenin levels were decreased and HUVECs motility was inhibited. Conclusions: TAZ may promote angiogenesis in gastric cancer by promoting β-catenin expression.

One major strategy to generate genetically modified mouse models is gene targeting in mouse embryonic stem (ES) cells, which is used to produce gene-targeted mice for wide applications in biomedicine. However, a major bottleneck in this approach is that the robustness of germline transmission of gene-targeted ES cells can be significantly reduced by their genetic and epigenetic instability after long-term culturing, which impairs the efficiency and robustness of mouse model generation. Recently, we have established a new type of pluripotent cells termed extended pluripotent stem (EPS) cells, which have superior developmental potency and robust germline competence compared to conventional mouse ES cells. In this study, we demonstrate that mouse EPS cells well maintain developmental potency and genetic stability after long-term passage. Based on gene targeting in mouse EPS cells, we established a new approach to directly and rapidly generate gene-targeted mouse models through tetraploid complementation, which could be accomplished in approximately 2 months. Importantly, using this approach, we successfully constructed mouse models in which the human interleukin 3 (IL3) or interleukin 6 (IL6) gene was knocked into its corresponding locus in the mouse genome. Our study demonstrates the feasibility of using mouse EPS cells to rapidly generate mouse models by gene targeting, which have great application potential in biomedical research.

Objective To investigate the differences between indocyanine green (ICG) fluorescence imaging plus methylene blue and plus Carbon Nanoparticles Suspension Injection for sentinel lymph node biopsy (SLNB)in breast cancer patients. Methods A total of 134 cases of early breast cancer patients performed SLNB from November 2013 to November 2016 were involved,of which 48 cases were performed with ICG fluorescence imaging plus methylene blue,and another 86 cases plus Carbon Nanoparticles Suspension Injection. Results There was no significant difference between ICG plus Methylene Blue group and ICG plus nano carbon group in terms of detection rate(P>0.05),detected numbers(P>0.05),sensitivity(P>0.05),accuracy(P>0.05)and false negative rate(P > 0.05). Age,and body mass index(BMI)exerted no influence on the detection rate and accuracy of SLNB in two groups(P>0.05). Conclusion ICG Fluorescence Imaging plus Methylene Blue showed similar detection rate , detected numbers , sensitivity , accuracy and false negative rate as it plus Carbon Nanoparticles Suspension Injection for SLNB in breast cancer patients ,and both of them can be performed easily and conveniently.

Objective To evaluate the efficacy and safety of donepezil in the treatment of cognitive dysfunction caused by radiation-induced encephalopathy.Methods A total of fifty-five patients with radiation-induced cognitive impairment were divided into treatment group with extra donepezil 5-10 mg/d combined with conventional therapy and control group with conventional treatment for 16 weeks.The cognitive function was assessed according to Montreal cognitive assessment (MoCA) and mini-mental state examination (MMSE) before and 16 weeks after treatment.Results After 16 weeks of treatment,the patients in treatment group displayed significantly greater improvement in cognitive function.In treatment group,the scores of patients after donepezil therapy in MoCA and MMSE were obviously higher than the control group (t =5.40,3.88,P ＜ 0.01).The scores in the visual space and executive function,naming,attention,abstract thinking,delayed memories also had improved,which suggested the statistically significant difference(t=-3.55,-3.08,-3.21,-2.58,-3.65,P＜0.05).The scores of control group unchanged accordingly.Conclusions Donepezil combined with conventional treatment was signally effective in the therapy of cognitive dysfunction caused by radiation-induced encephalopathy.