Objective: To prepare and optimize tilianin loaded solid lipid nanoparticles (T-SLNs), and investigate the physicochemical properties, absorption and transport behavour of T-SLNs in vitro. Methods: T-SLNs were prepared by high shear homogenization followed by ultrasonication and optimized by central composite design and response surface methodology. In the study, the physicochemical properties of T-SLNs including size, polydispersity (PDI), Zeta potential, shape, entrapment efficiency and release profile in vitro were investigated, the absorption and transport behavour of T-SLNs in Caco-2 cell model were also measured. Results: The optimum formulation of T-SLNs consisted of: drug/lipid of 0.11, soy lecithin/lipid of 1.26, and content of tween-80 was 5.05%. The prepared T-SLNs were spherical and uniform with the mean particle diameter at (86.40 ± 0.62) nm, PDI (0.165 ± 0.080) and Zeta potential of (-24.2 ± 0.6) mV, respectively. The average EE was (89.81 ± 1.07)%, and the release in vitro showed that tilianin was released about (98.72 ± 1.57)% in 48 h. Besides, the absorption and transport assays of T-SLNs in Caco-2 cells model indicated that T-SLNs had a higher absorption and transport than tilianin. Conclusion: The method of high shear homogenization followed by ultrasonication is suitable for T-SLNs preparation. The optimal T-SLNs have a smaller particle size and high EE. Moreover, in the same concentration of tilianin, the absorption and transport amounts of T-SLNs in Caco-2 cell model were higher than tilianin.

Objective To investigate the consistency of the anatomical parameters of left atrial appendage (LAA) based on three-dimensional transesophageal echocardiography (3D-TEE) and CT of 3D print datasource by the post-processing of DICOM data.Methods Sixty-three patients with atrial fibrillation in our hospital who underwent 3D-TEE and cardiac CT examination were selected and the original DICOM data of the LAA were obtained.The volume images of LAA were acquired by the postprocessing of Mimics software.After measuring and evaluating the anatomical parameters,the differences of parameters between TEE and CT were compared and the consistency of the two methods was evaluated.Results The data of 63 patients with atrial fibrillation obtained by 3D-TEE and CT were successfully post-treated and the detailed anatomical parameters of LAA were obtained.Morphological parameters:the Kappa values of the anatomical shape and opening shape of the LAA based on the 3D-TEE and CT data were 91.0%,69.3%,respectively.Measurement parameters:the area,perimeter,long axis,short axis of ostiums of LAA,and depth of LAA by CT were larger than those by 3D-TEE(all P<0.05).The post-processing results showed that 57 patients (90.5%) had significant bending of LAA′s main lobe in the 3D-TEE group and CT group,the difference in the measurements of the angle of the first bend and the distance from the first bend of the LAA to the orifice of the 57 cases between by 3D-TEE and CT were no statistical significance (P>0.05).LAA′measurements of 3D-TEE was concordant well with CT′s by Bland-Altman analysis.Conclusions In agreement with CT,3D-TEE can also be used as a datasource for 3D printing by evaluating spatial morphology of the LAA.

Objective To evaluate the left ventricular diastolic function in patients with uremia by two-dimensional speckle tracking technology ( 2D-STI ) . Methods Ninety-three patients with uremia ( serum creatinine value≥ 700 μmol/l) and 29 controls were included . Uremia patients were divided into group with decreased LVEF ( LVEF < 50％ ,group C ,19 cases) and group with normal LVEF ( LVEF≥50％ ) . The latter were divided into group with normal diastolic function ( group A ,31 cases) and group with diastolic dysfunction ( group B ,23 cases) according to the diastolic function;pulsed wave tissue Dopper imaging datas included:diastolic velocity e1′,a1′ and e2′,a2′ at septal or lateral sites of mitral annulus , average E/e′and e′/a′. 2D-STI strain and strain rate parameters included:the longitudinal peak systolic strain ( GLS) ,the longitudinal peak systolic strain rate ( LSRs) ,longitudinal peak strain rate in early diastole (LSRe)andE/LSRe.Results ①Systolicfunctionparameters:comparedwithcontrolgroup,LVEF decreased significantly only in group C( P <0 .05) ,while there was no significant difference among group A ,B and control group ( P> 0 .05 ) ;GLS in group A ,B and C decreased significantly and showed a decreasing trend among these three groups( P <0 .05) ;LSRs in group B and C decreased significantly ,while there was no significant difference between group A and control group( P > 0 .05) . ② Diastolic function parameters:compared with control group ,the average E/e′decreased significantly in group B and C( P <0 .05) ,while group A showed no significant difference( P > 0 .05) . Compared with group A ,the average E/e′increased significantly in group B and C ( P < 0 .05) . Compared with control group ,LSRe in three uremia groups decreased significantly ,which showed a decreasing trend among these three groups ( P <0 .05) ;however ,E/LSRe in three uremia groups increased significantly and showed an increasing trend among these three groups ( P < 0 .05 ) . ③ The correlation analysis showed that there were positive correlations between average e′ and LSRe and between average E/e′ and E/LSRe in uremic patients . Conclusions In patients with uremia ,left ventricular systolic and diastolic function impaired in the early phase . Strain rate parameters in diastole especially E/LSRe obtained by 2D-STI can be used as sensitive , early parameter for evaluation of left ventricular diastolic dysfunction .

OBJECTIVETo investigate the ability of Porphyromonas gingivalis to invade human periodontal ligament cells (hPDLCs) and the effect of intracellular P. gingivalis on cell proliferation and osteogenic differentiation in vitro.

METHODSThe invasion ability of P. gingivalis in hPDLCs was tested using an antibiotic protection assay at the multiplicity of infection (MOI) of 10 and 100. The proliferation of the infected cells was detected using a CFDA-SE kit, and the cells were sorted by fluorescence-activated cell sorting (FACS) followed by alizarin red staining for detecting mineralization nodules deposition; real-time PCR was used to examine the expression of Runx2 mRNA in the cells.

RESULTSP. gingivalis actively invaded hPDLCs, and the internalized P. gingivalis was able to resist antibiotic treatment. The cells infected by P. gingivalis exhibited no significant suppression of cell proliferation, but showed significantly lowered capacity for osteogenic differentiation, down-regulated RUNX2 mRNA expression, and reduced mineral deposition.

CONCLUSIONIntracellular P. gingivalis does not significantly affect the proliferation of hPDLCs but inhibits osteogenic differentiation of the cells.

Cell Differentiation ; Cell Proliferation ; Cells, Cultured ; Core Binding Factor Alpha 1 Subunit ; metabolism ; Flow Cytometry ; Fluoresceins ; Humans ; Osteogenesis ; Periodontal Ligament ; cytology ; microbiology ; Porphyromonas gingivalis ; RNA, Messenger ; Real-Time Polymerase Chain Reaction ; Succinimides

OBJECTIVETo explore an experimental method for construction of tissue-engineered heart valve (TEHV) in canine abdominal aorta.

METHODSThe decellular porcine aortic valve (PAV) leaflets seeded with canine vessel interstitial cells and endothelial cells (ECs) were implanted into 6 canine abdominal aortas. Valve specimens were obtained respectively at the end of 4, 6, 8 and 10 weeks after implantation were studied for morphology, histology and immunohistochemistry.

RESULTS(1) After 4 weeks implantation, multiple layers of cells grew into peripheral portion of valve scaffold, while new extracellular matrix appeared, and original scaffold tissue was partially absorbed. (2) At the end of 10th week after implantation, the decellular PAV scaffold disappeared completely and was substituted by recipient cells and new extracellular matrix. The interstitial cells in matrix was mainly consisted of fibroblasts and myofibroblast. The matrix was mainly composed by type I, III collagen, some elastic fibers with neutral and acid mucopolysaccharide. (3) Surface of valve leaflets were covered with endothelial cells.

CONCLUSIONS(1) TEHV is primarily constructed with recellularized PAV after implantation into canine abdominal aorta for 10 weeks. (2) Heterotopic implantation into the abdominal aorta is an alternative experimental procedure to study the TEHV.

Animals ; Aorta, Abdominal ; surgery ; Aortic Valve ; cytology ; transplantation ; Bioprosthesis ; Dogs ; Heart Valve Prosthesis ; Male ; Prosthesis Design ; Swine ; Tissue Engineering ; methods ; Transplantation, Heterologous

OBJECTIVETo investigate the effects of prebiotics supplementation for 9 days on gut microbiota structure and function and establish a machine learning model based on the initial gut microbiota data for predicting the variation of Bifidobacterium after prebiotic intake.

METHODSWith a randomized double-blind self-controlled design, 35 healthy volunteers were asked to consume fructo-oligosaccharides (FOS) or galacto-oligosaccharides (GOS) for 9 days (16 g per day). 16S rRNA gene high-throughput sequencing was performed to investigate the changes of gut microbiota after prebiotics intake. PICRUSt was used to infer the differences between the functional modules of the bacterial communities. Random forest model based on the initial gut microbiota data was used to identify the changes in Bifidobacterium after 5 days of prebiotic intake and then to build a continuous index to predict the changes of Bifidobacterium. The data of fecal samples collected after 9 days of GOS intervention were used to validate the model.

RESULTSFecal samples analysis with QIIME revealed that FOS intervention for 5 days reduced the intestinal flora alpha diversity, which rebounded on day 9; in GOS group, gut microbiota alpha diversity decreased progressively during the intervention. Neither FOS nor GOS supplement caused significant changes in β diversity of gut microbiota. The area under the curve (AUC) of the prediction model was 89.6%. The continuous index could successfully predict the changes in Bifidobacterium (R=0.45, P=0.01), and the prediction accuracy was verified by the validation model (R=0.62, P=0.01).

CONCLUSIONShort-term prebiotics intervention can significantly decrease α-diversity of the intestinal flora. The machine learning model based on initial gut microbiota data can accurately predict the changes in Bifidobacterium, which sheds light on personalized nutrition intervention and precise modulation of the intestinal flora.

To evaluate the right ventricular systolic function in uremia patients and the effects of maintenance hemodialysis on right ventricular systolic function by volume and strain parameters obtained by 4D RV Volume . Methods Seventy‐six patients with uremia and twenty‐two controls were selected . According to left ventricular ejection fraction ( LVEF) ,uremia patients were divided into normal LVEF uremia group and decreased LVEF uremia group . T hen normal LVEF uremia group was divided into maintenance hemodialysis group and non‐dialysis group . Conventional ultrasound parameters included :LVEF ,pulmonary artery systolic pressure ( PASP) and tricuspid annular plane systolic excursion ( T APSE‐2D) . 4D RV Volume parameters included : right ventricular end‐diastolic volume ( RVEDV ) , tricuspid annular plane systolic excursion ( T APSE‐4D ) ,right ventricular area change rate ( FAC ) ,right ventricular ejection fraction ( RVEF) and right ventricular free wall longitudinal systolic strain ( RV‐GLSfree ) . Results①Compared with the control group ,T APSE‐2D decreased significantly in the decreased LVEF uremia group ( P ＜0 .05) ,w hile there was no significant difference of T APSE‐2D in normal LVEF uremia group ( P ＞ 0 .05) . Compared with the control group and normal LVEF uremia group ,PASP increased significantly in the decreased LVEF uremia group ( P ＜ 0 .05 ) . Compared with the control group ,RVEDV increased significantly both in the normal LVEF and decreased LVEF uremia group ,w hich showed an increasing trend in these three groups ( P ＜0 .05) ,while T APSE‐4D ,FAC ,RVEF and RV‐GLSfree all decreased significantly and showed a decreasing trend in these three groups ( P ＜ 0 .05 ) . ② Compared with the control group , T APSE‐2D decreased significantly in non‐dialysis group ( P ＜0 .05) ,but there was no significant difference in uremia hemodialysis group ( P ＞0 .05) . Compared with the control group ,PASP and RVEDV increased and T APSE‐4D ,FAC ,RVEF and RV‐GLSfree decreased significantly in uremia hemodialysis group and non‐dialysis group ( P ＜ 0 .05 ) . Compared with non‐dialysis group , T APSE‐2D and T APSE‐4D increased significantly in hemodialysis group ( P ＜0 .05) ,while there was no significant difference in RVEDV ,FAC , RVEF and RV‐GLSfree in uremia hemodialysis group ( P ＞0 .05) . Conclusions 4D RV Volume could early and accurately evaluate the right ventricular systolic dysfunction in uremia patients . Furthermore ,w hen evaluating right ventricular systolic function in uremia patients treated with maintenance hemodialysis , indices such as right ventricular strain and volume parameters should be comprehensively considered .

OBJECTIVETo characterize the incidence, epidemiologic features, etiologic agents and sequelae of bacterial meningitis in children under 5 years of age in Nanning, Guangxi.

METHODSA population-based surveillance was conducted to evaluate children with signs and symptoms of meningitis. All hospitals, township health centers and village clinics in the surveillance area were structured to participate in the case referral and evaluation. Cerebrospinal fluid (CSF) and blood specimens were obtained and processed using standardized microbiologic methods.

RESULTSDuring the 26-month surveillance period, among the children under 5 years old, a total of 1272 cases who met the screening criteria of meningitis were studied. 265 of 1272 cases were identified as clinically diagnosed meningitis, with an incidence rate of 86.36 per 100 000 population. The annual incidence rate under the 38 cases of confirmed bacterial meningitis appeared to be 12.38/100 000. Staphylococcus species accounted for the largest proportion of laboratory-confirmed bacterial meningitis, followed by E. coli and S. pneumoniae. The highest attack rate occurred in neonates < 1 month, followed by children aged 1 - 12 months in the confirmed patients. Meningitis caused by Sp and Hi mainly occurred in children aged 1 - 12 months. All cases of meningitis due to Hi and Sp were children aged 1 - 24 months. 13.16% and 0.00% of the cases survived with complications and sequelae, and the case-fatality rate was 18.42%. 40 bacterial isolates were identified from 1193 blood cultures and 23 from 1211 cerebrospinal fluid samples, but no Neisseria meningitidis was found.

CONCLUSIONMeningitis due to Hi was first confirmed in Guangxi with the incidence of 0.98 per 100 000 population. The annual incidence rate of confirmed bacterial meningitis was 12.38 per 100 000, which was considered an important public health problem in children. Staphylococci was the predominant pathogen in confirmed bacterial meningitis.

Child, Preschool ; China ; epidemiology ; Female ; Humans ; Incidence ; Infant ; Infant, Newborn ; Male ; Meningitis, Bacterial ; epidemiology ; microbiology ; Meningitis, Escherichia coli ; epidemiology ; Meningitis, Haemophilus ; epidemiology ; Population Surveillance ; Staphylococcal Infections ; epidemiology

Growing Up in Singapore Towards healthy Outcomes (GUSTO) is Singapore's largest birth cohort study to date. The main aim of GUSTO is to evaluate the role of developmental factors in the early pathways to metabolic compromise. Detailed data is collected for a range of environmental exposures in the parents and offspring, and allergic disorders are among a number of outcomes assessed in infancy and childhood. Under the Allergy domain of GUSTO, this integrated study will describe the epidemiology of allergic manifestations and different phenotypes in the Asian context and help shed light on the association of metabolic disease to allergy. Epigenetic mechanisms and associations with other childhood disorders will also be explored. The aim of this report is to focus on methodology of GUSTO, and to suggest similar approaches (i.e., integrated cohort studies on pediatric allergy) worldwide. Recruitment commenced in 2009 with a cohort of 1,163 pregnant mothers in their first trimester. The mothers and children were followed throughout pregnancy and follow-up will continue until the child reaches 3 years of age. Preliminary results showed that 39.8% of the mothers had a personal history of having at least one allergic disease, which included asthma, eczema and allergic rhinitis. Further data collection and analyses are still ongoing. Allergy is a complex spectrum of disorders with numerous poorly-understood aspects. The ongoing GUSTO cohort study, with its longitudinal design and multi-disciplinary nature, may provide new insights into developmental influences on allergy. As a Singapore-based study, it will be the first integrated allergy cohort in Southeast Asia, of which recruitment started during pregnancy.
Asia, Southeastern ; Asian Continental Ancestry Group ; Asthma ; Child ; Cohort Studies ; Data Collection ; Eczema ; Environmental Exposure ; Epidemiology ; Epigenomics ; Female ; Follow-Up Studies ; Humans ; Hypersensitivity ; Metabolic Diseases ; Mothers ; Parents ; Parturition ; Phenotype ; Pregnancy ; Pregnancy Trimester, First ; Rhinitis, Allergic ; Singapore

OBJECTIVETo explore the feasibility of using next-generation sequencing technology (NGS) to screen the neonatal thalassemia genes.

METHODSPlantar blood of 206 cases of neonatal born in our hospital were randomly collected to be made into dried blood, which can be screened for thalassemia genes by next-generation sequencing, and then a further analysis would be performed on the basis on the detection results.

RESULTSIn 206 cases of neonates tested, the thalassemia gene mutations in 22 cases were screened, including 11 cases of alpha-thalassemia, 11 cases of beta-thalassemia, 5 cases of new mutations. Out of 11 cases of alpha-thalassemia 7 cases were proved to be the gene deletion, accounting for 64% (7/11), and the specific genotype distribution was as follows: 4 cases of αα/-α(3.7), 2 cases of αα/-SEA, 1 case of αα/-α(4.2), the remaining 4 cases with point mutations (4/11, 36%): Hb Part-Dieu hybrid, Hb Quong Sze hybrid, Hb Westmead hybrid, HBA1: c. 95 + 9 c > T (rewly discovered gene mutation). The whole 11 cases of β-thalassemia are proved to be with beta chain point mutations, 7 kinds of mutation genotype were detected , CD17 (A->T) is the most common point locus mutation, accounted for 27% (3/11), and 50 G>A hybrid in 2 cases, 1 cases of Hb Hamilton hybrid, IVS-II-654 (C->T) in 1 case. The remaining 4 cases are of the new gene point mutation, they are as follows respectively: HBB: c. 316-116 c>A, HBB: c.316-248G>T, HBB: c.315 + 63 T>c, HBB: c. -23 A>G.

CONCLUSIONThe next-generation sequencing technology can be used to screen neonatal plantar dried blood for the thalassemia genetic mutation, which not only can effectively detect thalassemia gene types, but also can look for new gene mutations. The advantages of this method include easy collecting samples, precise result and wide use for clinical diagnosis, thus possibly give an early diagnosis for thalassemia.

DNA Mutational Analysis ; Gene Deletion ; Genotype ; Hemoglobins, Abnormal ; genetics ; High-Throughput Nucleotide Sequencing ; Humans ; Infant, Newborn ; Mutation ; Point Mutation ; alpha-Thalassemia ; genetics ; beta-Thalassemia ; genetics