【Objective】 To investigate the relationship between hepatitis B virus X （HBx） protein and EGFR promoter， and the role of HBx protein in activating EGFR/PI3K/p-Akt signaling pathway and inhibiting apoptosis. 【Methods】 EGFR promoter plasmids were constructed and the relationship between HBx and EGFR promoters was characterized using a luciferase reporter assay. EGFR-overexpressing trophoblast cells were constructed， and EGFR expression in the overexpressing cells was knocked down using EGFR shRNA. The expression and localization of EGFR/PI3K/p-Akt were detected by Western blotting and confocal laser microscopy. Cell apoptosis was analyzed using flow cytometry. HBV plasmids carrying either full-length HBx or HBx with a deletion mutation （ΔHBx） and HBx plasmids were transfected into two types of trophoblast cells； HBx and PI3K/p-Akt protein expressions were detected by Western blotting. Cell apoptosis was analyzed using flow cytometry. 【Results】 Co-transfection of HBx and EGFR promoter plasmids in JEG-3 and HTR-8/Svneo cells significantly elevated the expression of EGFR promoter driven luciferase compared with the control group （P<0.01）. In EGFR-overexpressing cells， the expression of PI3K/p-Akt was significantly increased （P<0.01）， whereas the apoptosis rate was significantly decreased for JEG-3 cells and HTR-8/Svneo cells （both P<0.01）. These results were reversed in the EGFR-knock down group. When the intracellular HBx protein was expressed in JEG-3 and HTR-8 cells， PI3K/p-Akt protein expression was significantly increased （both P<0.05）， and the proportion of apoptosis was significantly decreased （both P<0.05）. 【Conclusion】 In placental trophoblast cells， HBx protein activates the expression of EGFR by acting on the EGFR promoter， and inhibits the apoptosis of trophoblast cells via the downstream EGFR/PI3K/p-Akt signaling pathway.

Objective To explore the association between the GRIN3B gene and Tourette syndrome (TS) in children by screening mutations in the coding region of this gene.Methods Fifty-one children with TS and their parents in the Affiliated Hospital of Qingdao University from October 2015 to November 2016 were selected as an experimental group,41 cases of boys,and 10 cases of girls,aged 6-16 years[(9.78 ±3.64)years],while 60 people aged 22-45 years in the health examination center were selected in the control group,49 cases were male,1 1 cases were female,aged 22-45 years [(29.08 ± 2.89) years].DNA was extracted from 51 patients with TS,their parents and 60 controls.PCR was applied to amplify the encoding region of GRIN3B gene and Sanger sequencing was used to sequence,then GRIN3B sequencing results were compared with the NCBI gene encoding region sequence (NM_138690.2)to test whether these patients carried gene mutation and to verify the findings from their family.Results c.C460T gene variant of GRIN3B was found in 2 patients (p.P154S);c.T1187C (p.L396S) variant of GRIN3B gene was found in 10 patients and both of abnormal GRIN3B sites lead to changes in amino acid.The 2 peak sequencing maps were obtained by Sanger sequencing but nothing was found in their parents.Conclusion The mutation of GRIN3B gene may be related to the development of TS.

Objective To explore the feasibility and safety of endovascular directional atherectomy angioplasty for the treatment of severe vertebral artery stenosis.Methods Directional atherectomy combined with use of drug-coating balloon (DCB) was employed to treat one patient with severe stenosis of vertebral artery in November 2017 at authors' hospital.Protective umbrella filter was placed at the distal site of V1 segment during the procedure course.Results The operation was successfully accomplished.No postoperative complications occurred.After the operation,the clinical symptom of dizziness disappeared and the patient was well recovered.Conclusion For the treatment of severe vertebral artery stenosis,directional atherectomy combined with use of DCB is safe and feasible.

Objective: To investigate the effect and significance of GSK-3β inhibitor(LiCl)and RANK-RANKL on the renal tissue of diabetic nephropathy(DN) rats. Methods: SD rats were divided into normal control group (NC), DN model group (DN) and GSK-3β inhibitor intervention group (LiCl). Twenty-four hour urine protein of rats were determined by Coomassie brilliant blue. Kidney tissue sections were stained by HE. The expression of GSK-3β, RANK and RANKL protein were determined by immunohistochemistry staining. The mRNA of GSK-3β, RANK, RANKL was detected by RT-qPCR. Results: Compared with NC group[(14.72±3.37)g], the level of 24-hour urinary protein[(154.17±20.65)g] increased significantly in DN group; compared with DN Group, the level of 24-hour urinary protein [(107.22±31.15)g]decreased in LiCl group(P<0.05). Compared with NC group(2.10±0.60, 1.10±0.20, 1.21±0.20; 19.52±3.20, 1.80±1.10, 1.81±0.50), the pathological changes of renal tissues of DN group aggravated, the mRNA and expression of protein of GSK-3β, RANK and RANKL increased(9.10±2.15, 8.95±2.40, 9.90±2.60; 32.70±7.20, 19.20±4.32, 20.92±5.90); compared with DN group, the pathological changes of renal tissues of LiCl group alleviated, mRNA and the expression of protein of factors above declined(2.70±0.80, 2.32±0.65, 3.58±1.10; 22.35±3.25, 4.20±2.42, 5.90±2.36; P<0.05). Conclusion RANK and RANKL play an important role in the development of DN, LiCl influence Wnt and NF-κB signal pathway down-regulating RANK and RANKL to suspend development of diabetic nephropathy.

Objective To explore the clinical effectiveness and safety of selective usage of embolic protection device to prevent distal embolization during SilverHawk atherectomy for atherosclerotic femoropo-pliteal artery disease. Methods From Jan 2014 to December 2015, 45 femoropopliteal artery atherosclerot-ic patients were treated with SilverHawk atherectomy and selective embolic protection device (EPD). The indication for EPD was instent restenosis, highly calcified lesion, suspicious of thrombosis, ulcerated le-sion, and single below-the-knee runoff. All cases who met the indication were treated with atherectomy and EPD, and those who did not meet the indication were treated with or without EPD according to the patient's choice. The embolic related complications were analyzed. Results Twenty three out of 45 patients who met the EPD indication were all treated with SilverHawk atherectomy under EPD protection, filter captured deb-ris in 17 patients (73. 9％) of the patients. The other 22 patients who did not meet the indication were di-vided into 2 groups according to the patient's choice of EPD usage, 11 were treated by atherectomy with EPD and 11 without EPD. One case out of 11 unindicated patients without EPD suffered a tibioperoneal trunk embolization and restored with catheter aspiration. For 1/11 (9. 1％) unindicated cases with EPD protec-tion, the filter captured embolization. There was a significant difference of distal embolization rate between the indicated and unindicated patients (χ2 =19. 368,P =0. 000). All filters were retrieved successfully without any distal embolization and any complications except arterial spasm occurred in 2 patients and re-stored well with nitroglycerin. Conclusions It is safe and effective for selective usage of embolic protection device to prevent distal embolization during SilverHawk atherectomy for atherosclerotic femoropopliteal artery disease.

Objective To investigate the clinical outcome of drug-coated balloon ( DCB) treated atherosclerosis obliterans ( ASO) in lower extremity. Methods Data of 28 patients were retrospectively an-alyzed to determine the effectiveness and characteristics of DCB treatment. Results All the 28 patients were successfully treated with endovascular intervention therapy. Lesions mainly located in the superficial femoral arteries were divided into Groups A, B, C, and D according to TransAtlantic InterSociety Consensus (TASC) Ⅱ classification. Follow-up at 6, 9, and 12 months showed 100％, 84. 6％ and 76. 9％ patency rate in treated artery. After 9 months of endovascular intervention therapy, the combined artery patency rates in Groups A and B was 92. 8％, which was significantly better than 66. 7％ in Groups C and D ( P <0. 05 ) . DCB angioplasty had an artery patency rate of 86. 7％, while DCB angioplasty plus stenting had the patency rate of 85. 7％ (P>0. 05). Artery patency rate of Simple DCB angioplasty in ten patients (Exclu-ding patients with in-stent restenosis ) was 80％ and of Atherectomy plus DCB angioplasty in 5 patients was 100％ (P<0. 05). Conclusions DCB angioplasty has superiority over conventional intervention therapy. Combined debulky and DCB angioplasty appear to be the best choice in current treatment of ASO in lower extremity.

Objective To investigate the application value of fetal umbilical cord blood hemoglobin analysis in the prenatal diag ‐nosis of thalassemia .Methods 113 couples were the carriers of the same gene type of thalassemia ,moreover the females were in the pregnant period of 24 - 30 pregnant weeks and performed the prenatal diagnosis .The fetal umbilical cord blood hemoglobin compo‐nents were analyzed by the full automatic capillary electrophoresis technique ,meanwhile the fetal thalassemia gene was detected .Re‐sults Among 113 fetuses ,the umbilical cord blood HbBart′s level in 11 cases of severe α thalassemia was 85 .0% - 95 .5% ,which in 9 cases of intermediate type α thalassemia was 22 .0% - 39 .5% ;the umbilical cord blood HbA level in 6 cases of severe β thalas‐semia was 0% - 0 .4% ,which in 17 cases of light type β thalassemia was 2 .1% - 12 .5% .Conclusion The fetal umbilical cord blood hemoglobin analysis could be used for rapid prenatal diagnosis of severe α ,β and intermediate type α thalassemia ,which can serve as a supplementary method for the prenatal diagnosis of thalassemia .

OBJECTIVEDiagnosis and prenatal diagnosis to a family of hemoglobin variant with α-thalassemia.

METHODSWhole blood cell analysis, hemoglobin analysis by capillary zone electrophoresis (CZE), Gap-PCR, polymerase chain reaction-reverse dot blot (PCR-RDB) assay and DNA sequencing.

RESULTSHb Zurich Albisrieden with α°-thalassemia lead to severe anemia. The genotype of fetus is also Hb Zurich Albisrieden with α°-thalassemia.

CONCLUSIONAbnormal hemoglobin with α-thalassemia may lead to severe anemia, Prenatal diagnosis of thalassemia has the vital significance for eugenic birth.

Adult ; Base Sequence ; Child, Preschool ; Female ; Fetal Diseases ; blood ; diagnosis ; genetics ; Hemoglobins, Abnormal ; genetics ; metabolism ; Humans ; Male ; Molecular Sequence Data ; Pregnancy ; Prenatal Diagnosis ; Young Adult ; alpha-Thalassemia ; blood ; diagnosis ; embryology ; genetics

Objective To study the diagnostic utility of hemoglobin electrophoresis in neonatal cord blood screening for thalassemia. Methods Between January 2012 and December 2013, 14032 core blood samples which were from different 21 Women and Children Hospitals in Guangdong were performed for the neonatal screening with hemoglobin electrophoresis. The positive samples of hemoglobin electrophoresis were recalled for genetic testing. Results Out of 1445 (11.07%) positive samples of hemoglobin electrophoresis , 1075 (54.08%) cases were suspected for α-thalassemia, 478 (3.41%) cases were suspected for β-thalassemia, 127 (0.91%) cases were suspected for abnormal hemoglobin. With the genetic testing, 967 cases were diagnosed as α-thalassemia, 404 cases were diagnosed asβ-thalassemia. The coincidence rate ofα-thalassemia andβ-thalassemia were 89.95%and 82.96%, respectively. Besides, 124 cases were diagnosed as abnormal hemoglobin, including 38 cases of Hb E, 28 cases of Hb Q, 21 cases of Hb D, 19 cases of Hb New York, 13 cases of Hb J, and 5 cases of Hb J. Conclusion Hemoglobin electrophoresis was definitely helpful in the neonatal cord blood screening for thalassemia and abnormal hemoglobin.