Objective:To explore the clinical and genetic etiology of a Chinese pedigree affected with type 2 Long QT syndrome (LQTS).Methods:A pedigree with type 2 LQTS presented at Fuwai Central China Cardiovascular Hospital on August 23, 2019 was selected as the study subject. Peripheral blood samples were collected from the proband and her parents. Following extraction of genomic DNA, whole exome sequencing (WES) was carried out for the proband, and candidate variant was screened through functional annotation and protein-protein interaction (PPI) analysis. Sanger sequencing was conducted to verify the pathogenicity of candidate variant. This study was approved by Medical Ethics Committee of the Fuwai Central China Cardiovascular Hospital (Ethics No. 2019-15).Results:WES revealed that the proband has harbored a missense variant of the KCNH2 gene, namely c. 1478A>G (p.Tyr493Cys), which was confirmed by Sanger sequencing to have inherited from her father. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the variant was classified as likely pathogenic (PM2_supporting+ PM5+ PP3+ PP4). Conclusion:The KCNH2 gene c. 1478A>G (p.Tyr493Cys) variant probably underlay the type 2 LQTS in this pedigree.

Objective:To investigate the influence of Kruppel-like factor 16 (KLF16) on the proliferation and migration of pancreatic cancer cells.Methods:Immunohistochemical images of KLF16 were collected from 171 pancreatic cancer tissues and their matched paracarcinoma normal pancreas tissues and 8 pancreatic cancer tissues only in GEPIA database. The expression of KLF16 protein was detected by immunohistochemical imaging software. The protein and mRNA expressions of pancreatic cancer cell lines AsPC-1 and MIA PaCa-2 KLF16 were detected by Western blot and quantitative fluorescence PCR. By knockdown or exogenous overexpression of KLF16, the two cells were divided into blank control group (NC group), negative control group (siRNA-NC group), downexpression KLF16 group (siKLF16 group), overexpression control group (OE-NC group) and ovexpression KLF16-OE group (KLF16-OE group). CCK-8 assay, colony formation assay and transwell chamber were used to detect cell proliferation and migration.Results:The KLF16 protein expression level (4.02±1.26 vs 1.73±1.07) and positive expression rate (91.6% vs 13.5%) in pancreatic cancer tissues were significantly higher than those in paracancer normal pancreas tissues, with statistical significance ( P<0.05). After downregulating KLF16 expression and culturing for 24, 48, 72, and 96 hours, the A450 values of both AsPC-1 (0.19±0.02 vs 0.23±0.03, 0.24±0.06 vs 0.36±0.06, 0.45±0.09 vs 0.78±0.10, 0.69±0.04 vs 0.88±0.07) and MIA PaCa-2 cells (0.20±0.03 vs 0.22±0.02, 0.29±0.05 vs 0.31±0.04, 0.47±0.06 vs 0.78±0.10, 0.71±0.02 vs 0.90±0.07) and colony counts [(36±4.32) per well vs (118.51±10.01) per well, (13.6±2.62) per well vs (83.1±9.11) per well], and the number of migrated cells [(16.67±2.05) vs (46.67±5.91), (19.67±1.69) vs (55±4.89)] all decreased significantly. However, after up-regulating the expression of KLF16 and culturing for 24, 48, 72 and 96 h, the A450 value of both AsPC-1 (0.21±0.05 vs 0.20±0.04, 0.48±0.03 vs 0.31±0.04, 0.91±0.09 vs 0.72±0.03, 1.28±0.10 vs 1.05±0.02) and MIA PaCa-2 cells (0.20±0.01 vs 0.19±0.05, 0.44±0.03 vs 0.30±0.04, 0.89±0.06 vs 0.72±0.03, 1.19±0.05 vs 1.01±0.10), and the number of cell colonies [(189±6.37)/per hole vs (108±9.62)/ per hole, (141±12.56)/ per hole vs (80.69±10.32)/ per hole]], migration cell numbers [(79±4.89) per hole vs (50.33±4.11) per hole, (79.66±3.85) per hole vs (51±4.08) per hole] all increased significantly. Conclusions:KLF16 is highly expressed in pancreatic cancer. The up-regulated expression of KLF16 in pancreatic cancer cell lines can promote the proliferation and migration of pancreatic cancer cells.

OBJECTIVE To optimize the preparation technology of ethanol extracts from Centipeda minima， and investigate the anti-inflammatory activities of different extraction sites. METHODS Single factor test and response surface methodology were adopted to investigate the effects of ethanol volume fraction， extraction time， solid-liquid ratio and extraction times on the heating reflux extraction technology of total triterpenoids ethanol extract using the extraction rate of total triterpenoids of C. minima as indexes， optimize the extraction technology and carry out validation. Using dexamethasone as positive control drug， the effects of different extraction sites of C. minima （petroleum ether part， ethyl acetate part， n-butanol part， water part） on nitric oxide （NO） production in mononuclear macrophage RAW 264.7 cells of mice induced by lipopolysaccharide （LPS） were compared； the half inhibitory concentration （IC50） was calculated. RESULTS The optimal extraction technology of total triterpenoids ethanol extracts of C. minima was as follows： ethanol volume fraction of 70%， solid-liquid ratio of 1∶40 （g/mL）， extraction time of 2.0 h and extraction times of 3 times. The 3 times of validation tests showed that average extraction rate of total triterpenoids of C. minima was 1.134%， relative error of which with the predicted value was 0.02%. The petroleum ether part and ethyl acetate part of C. minima could inhibit the generation of NO in RAW 264.7 cells induced by lipopolysaccharide to different degrees. IC50 values of NO production were 2.44 μg/mL and 2.22 μg/mL， respectively， and both of them were lower than those of positive control drug dexamethasone （7.65 μg/mL）. CONCLUSIONS The optimized preparation process of ethanol extracts from C. minima is stability and feasibility. The petroleum ether part and ethyl acetate part have obvious anti-inflammatory effects.

Objective:To identify and characterize two Balneatrix alpica strains isolated from a patient′s blood sample (strain X117) and the natural hot spring water in the patient′s residential district (strain GN-1), and to provide experimental evidence for the pathogenic diagnosis of clinical infection caused by this rare pathogen. Methods:Biochemical phenotypic identification, matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS), 16S rRNA gene sequencing, phylogenetic analysis, single-nucleotide polymorphism (SNP) analysis, and genome-wide analysis were performed to accurately determine the taxonomic status of the isolates X117 and GN-1 by using Balneatrix alpica DSM 16621 T as a reference. Microdilution broth method was used to test their antimicrobial susceptibility. The virulence genes carried by them were annotated and analyzed using the virulence factor database (VFDB). Results:Strains X117 and GN-1 formed light yellow or tan colonies with mottled surfaces on Columbia blood agar and chocolate agar plates after 4 d of culture. They were Gram-negative rods and positive for oxidase and indole tests, which were consistent with the characteristics of Balneatrix alpica DSM 16621 T. The phylogenetic analysis based on the 16S rRNA gene showed that the isolates X117 and GN-1 were both Balneatrix alpaca. The average nucleotide identity (ANI) values between the two isolates and Balneatrix alpica DSM 16621 T were 98.44% and 98.41%, respectively, and the digital DNA-DNA hybridization (dDDH) values were both 87.1%. The SNP distance between the two strains was 13, indicating that X117 and GN-1 might belong to the same clone. The antibiotic susceptibility testing showed that all of the three Balneatrix alpica strains were sensitive to the commonly used antibiotics against Gram-negative rods. The virulence genes carried by the three Balneatrix alpica strains were mainly involved in adhesion, invasion, flagella and biofilm formation. Conclusions:This study identified a case of bloodstream infection caused by Balneatrix alpica which was closely related to natural hot spring water. Natural hot spring water migh be an important source of clinical infections caused by this species.

Objective:To explore the risk factors and prognosis of lymph node metastasis and residual cancer following additional surgery after endoscopic submucosal dissection (ESD) in early gastric cancer (EGC).Methods:Retrospective analysis was performed on the data of 42 patients with EGC who received additional surgery after ESD in General Surgery Department of Beijing Friendship Hospital, Capital Medical University from August 2012 to August 2019, including 35 males and 7 females, with a male to female ratio of 5∶1 and an average age of 62 (32 to 82 years old). The primary outcomes were lymph node metastasis risk and residual cancer risk, while the secondary outcomes were 3-year and 5-year overall survival. Logistic regression model was used to analyze the risk factors of lymph node metastasis and residual cancer, and Kaplan-Meier survival analysis was performed.Results:Multivariate analysis showed that gender ( OR: 45.3, 95% CI: 3.762-546.250, P=0.003), invasion depth ( OR: 3.965, 95% CI: 1.1019-15.432, P=0.047) and histological type ( OR: 9.455, 95% CI: 0.946-94.482, P=0.049) were independent risk factors for lymph node metastasis of early gastric cancer. The type of tumor invasion ( OR: 10.675, 95% CI: 1.840-61.932, P=0.008) and the horizontal resection margin ( OR: 9.341, 95% CI: 1.47-59.346, P=0.018) were independent risk factors affecting the occurrence of residual cancer. Stratified analysis showed that in men, the tumor invaded to T 1b-SM1, and the pathological type was undifferentiated; and in women, the tumor invaded to T 1b-SM2, regardless of the pathological type, lymph node metastasis occurred. When the horizontal margin was positive, regardless of the infiltration pattern; and the infiltration pattern is INF-c, regardless of the horizontal margin, residual cancer occurred. Survival analysis showed that the 3-year (100% vs 60%, P< 0.001) and 5-year overall survival rate (100% vs 25%, P< 0.001) were better than those with lymph node metastasis.The 3-year (100% vs 80%) and 5-year overall survival rates (100% vs 62.5%) of patients without residual cancer were significantly better than those with residual cancer, with statistically significant differences ( P<0.001). Conclusions:Gender, invasion depth and histological type are independent risk factors for lymph node metastasis of early gastric cancer, and tumor invasion form and horizontal incision margin are independent risk factors for the appearance of residual cancer. The long-term survival rate of patients with early gastric cancer without lymph node metastasis and residual cancer was significantly better than that of patients with lymph node metastasis and residual cancer. Therefore, ESD is one of the safe and effective treatment methods for patients with early gastric cancer, but some patients need additional surgery according to the specific situation in order to improve the prognosis.

Objective To investigate the relationship between the levels of programmed death ligand 1 (PD-L1) on circulating B cells and disease severity as well as renal injury of patients with systemic lupus erythematosus (SLE) .Methods 50patients with SLE were enrolled from May 2015to May 2017, and 20healthy individuals were enrolled as the control group.All patients were scored for their disease severity and renal injury.The expression of PD-L1on circulating B cells were evaluated by flow cytometery.Results In the group of patients with active SLE, the percentage of CD19+in B cells was (11.35±3.65) ％and the percentage of in CD19+B cells was (2.47±0.89) ％, both of which were significantly higher than that in the group of patients with non-active SLE and control group (P<0.05) .In addition, patients with SLE were divided into lower group and higher group according to the median percentage of CD19+and CD19+/PD-L1Bcells.The incidence of kidney lesion was robustly increased in the high group, and CD19+/PD-L1Bcells, LNⅣwere independent risk factors of renal insufficiency in lupus nephritis.Conclusion The level of peripheral CD19+/PD-L1in SLE patients increased significantly and was closely related to the activity degree and renal injury of the disease, which was an independent risk factor for renal insufficiency in patients.

OBJECTIVE:To establish HPLC fingerprint of Miao medicine Fufukang spray. METHODS:The determination was performed on Thermo Hypersil GOLD C18 column with mobile phase consisted of acetonitrile-0.2％ glacial acetic acid (gradient elution) at the flow rate of 0.6 mL/min. The detection wavelength was set at 262 nm,and column temperature was 25 ℃. The sample size was 10 μL. Using 6′-hydroxyl-justicidin B as reference control,HPLC chromatograms of 10 batches of samples were drawn. Similarity evaluation system software(2004 A edition)of TCM fingerprint was used to evaluate the similarity and determine common peaks of 10 batches of samples. The established method was used to determine the contents of 6′-hydroxyl-justicidin B and justicidin B in 10 batches of samples. RESULTS:There were 12 common peaks in HPLC fingerprints of 10 batches of samples with similarity>0.90. HPLC fingerprint showed a good consistency with the control fingerprint. The contents of 6′-hydroxyl-justicidin B and justicidin B in 10 batches of samples were 1.64-5.84,0.83-2.78 mg/100 mL,respectively. CONCLUSIONS:The established method is simple,accurate,reliable and suitable for quality evaluation of Fufukang spray.

Background Lattice corneal dystrophy (LCD) is a progressive disease,whose clinical features are varied in different stages.It is rarely be reported that clinical findings of different stages and factors of promoting the occurrence and development on LCD in a family.Objective The aim of this study was to identify the characteristics of the pedigree and clinical features of different stages in a LCD family,and further to discuss its influence factors.Methods A cross-sectional study was performed in this study.A Chinese family with LCD was enrolled in Shenzhen Eye Hospital from 2015 to 2016.Questionnaires for disease-related history,visual acuity measurement,ocular anterior segment examination and color photography were carried out for all the members of the family.In addition,anterior segment OCT (AS-OCT),laser scanning confocal microscope and corneal endothelium microscope were used to observe the morphology of corneal stroma and changes of corneal endothelial cells.The pedigree chart was drawn by Cyrillic2.1 software and analyzed based on Mendel law.Results This family included 5 generations of 73 members.Patients with LCD were found in each generation with similar morbidity in different gender,which followed the law of autosomal dominant inheritance.Eleven patients were found in 49 members related with Ⅲ1 of this family with the prevalence rate of 22.45％ and onset age at 21-50 years old,and the course of disease was 3-34 years.All of the members had no systemic disease except for two patients (Ⅲ 1 and Ⅲ 5) with hypertension.In the early stage of LCD,some bifurcate striolae appeared in the patients' corneal stroma without symptoms for many years.In the progressive stage,there was corneal irritation symptom accompanying with vision's decrease in the eyes with LCD.The bifurcate striolae were increased,widened and interwoven into lattice lines that the boundaries gradually became fuzzy,then corneal macula was formed because of recurrent corneal infiltration,and eventually resulted in corneal leucoma.High reflection corresponding to the pathologic region was showed by laser scanning confocal microscope and AS-OCT.No significant differences were found in corneal endothelial cell density and the percentage of hexagonal cells between LCD patients and normal phenotype families (t =1.887,P=0.075;t=-0.719,P =0.481).Penetrating keratoplasty was performed in a patient with corneal opacity and serious corneal opacity occurred near the surgical incision one year after the surgery.One patient was diagnosed as LCD 2 years after laser assisted in-situ keratomileusis.One patient was a welder.Conclusions LCD is autosomal dominant inheritance in the family.The clinical manifestations of LCD in the early,progressive and late stage can be seen in the pedigree,which offers a reference for ophthahnologists.Corneal surgery and lesion may induce the onset or aggravation of LCD.

Background Posner-Schlossman syndrome (PSS) is often recurrent and is a cause of blindness.The etiology of PSS remains to be elucidated.It is reported that there is a certain association between pathogenic microorganisms and PSS in rather small samples.Objective This study was to analyze the related serum antibody levels of cytornegalovirus (CMV),herpes simplex virus (HSV),rubella virus (RV),helicobacter pylori (HP) and anti-streptolysin O (ASO) and provide a clue for the study on pathogenesis and therapy of PSS.Methods A prospective cases-controlled study was carried out in Shenzhen Eye Hospital from December,2014 to December,2016 under the approval of Ethic Committee of this hospital and informed consent of each subject prior to initial of any medical examination.Peripheral blood samples were collected from 82 PSS patients as the PSS group and 100 age-and gender-matched healthy blood donors as the normal control group.The positive rates of serum CMV IgG,CMV IgM,HSV IgG,HSV IgM,RV IgG,RV IgM,HP IgG and HP IgM in the subjects were detected by indirect ELISA,and the positive rate of serum ASO antibody was determined by immuno-scatter turbidmetry.Results The positive rates of serum CMV-IgG,CMV-IgM,HP-IgG,HP-IgM and ASO antibody were 22.0％,17.1％,22.0％,17.1％ and 17.1％ in the PSS group,which were significantly higher than 5.0％,0.0％,10.0％,2.0％ and 7.0％ in the normal control group (x2 =11.726,18.496,4.943,12.766,4.479,all at P＜0.05).The positive rates of serum HSV-IgG,HSV-IgM,RV-IgG and RV-IgM in the PSS group were not significantly different from those in the normal control group (x2 =3.305,0.986,0.898,0.503,all at P ＞ 0.05).Conclusions CMV,HP and hemolytic streptococcal infection may participate in the occurrence and development of PSS.

Objective To explore the features of coronary artery morphological changes and left ventricular function indicators of resident population in plateau .Methods 50 plateau healthy adults and 50 plain‐and Dalian‐healthy volunteers completed coronary CTA examination .We measured the pipe diameter of left main coronary atery and right coronary artery and their branches ,respec‐tively .Furthermore ,SV and EF value of left ventricular cardiac function indexes and the wall thickening of free ventricular wall at left ventricular end systolic were measured .Results The average diameter of proximal right coronary of plain‐and Dalian‐healthy volun‐teers thicker than plateau group ,with statistical difference ;Other coronary artery branch did not exhibit any significant between‐group differences ;The two groups did not show significant between‐group differences with heart function indexes .Conclusion The body circulation system have adaptability and compensatory changes of resident population in plateau ,and their coronary artery mor‐phology and left ventricular function are as similar as plain people .