Objective:Study the effect about cone beam CT(CBCT) image guidance sytem to improve the positioning accuracy and reducing the set-up uncertainty in precise radiotherapy. Methods:Use CBCT system to scan head and neck tumors patients (30 cases) and chest tumor patients (40 cases), including head and neck tumor patients were scaned 90 times, chest tumor patients were scaned 113 times. Setup deviation statistics about anterior and posterior, head and feet, left and right direction. Results:The patients with head and neck cancer, the maximum error of anterior-posterior is 6mm, Three directions errors which greater than 3mm all less than 10%. Patients with chest tumor position error are in the head and foot direction, greater than 5 mm up to 21.51%. Before and after the direction of the error did not exceed 5mm. The left and right direction greater than 5 mm up to 4.53%. Conclusion:It greatly enhances the precision of radiotherapy, improved the curative effect of radiotherapy by using CBCT online image guidance system for the patient position correction. Compared with EPID, patients scaned with CBCT absorb smaller cumulative dose, the image resolution is higher, image matching is more accurate. But compared with the ordinary helical CT,the resolution and scanning range also need to improve.

OBJECTIVETo isolate and identify human dental pulp stem cells from third molars.

METHODSDental pulps were dissected and digested by collagenase type I and dispase. The obtained single cell supernatant were harvested and cultured. Characterization of the phenotype of DPSCs was detected by immunohistochemical method and RT-PCR assay. Cell cycle was analyzed by FCM. Differentiation potential of DPSCs was evaluated.

RESULTSColony-forming efficiency of cells derived from dental pulp tissue was 2 - 15 clones/10(3) cells plated. DPSCs were found to express many different markers, including vimentin, collagen type I, GFAP, nestin and osteocalcin, while they failed to react with MyoD and DSPP. About 64.1% of the cells were in G0/G1 phases, while only 35.8% in proliferation (S + G2 + M). Grown in an adipogenic cocktail medium for three weeks, some DPSCs expressed fat cell markers of PPARgamma and LPL, and formed oil red O-positive lipid clusters in five weeks. After culture with a myogenic-inductive medium, DPSCs were found to express MyoD, desmin and myosin, markers of myocyte. Long-term cultures of DPSCs grown in differentiation inductive medium demonstrated the capacity to form Von Kossa-positive condensed nodules with high levels of calcium.

CONCLUSIONCells isolated from adult human dental pulp are clonogenic, and have multipotent differentiation potential, satisfying the criteria of postnatal somatic stem cell.

Adult ; Cell Differentiation ; Dental Pulp ; Epithelial Cells ; Humans ; Osteocalcin ; Stem Cells

Objective To investigate the effects of specific RNA interference (RNAi) to Notch ligand Delta1 on proliferation and differentiation of human dental pulp stem cells (DPSC).Methods DPSC were infected by lentivirus vectors carrying Delta1-RNAi.DPSC were divided into three groups,DPSC/Delta1-RNAi group,DPSC/wt group and DPSC/vector group as control Cell counting kit-8( CCK-8 ) assayand flow cytometry were used to evaluate the proliferation of DPSC.Expression of proliferating cell nuclear antigen (PCNA) was examined with immunohistochemical staining.All groups were cultured in an odonto-inductive medium and were observed under microscope.The number of mineralization nodules was counted after Alizarin red staining.Alkaline phosphatase (ALP) activity and the expression of dentin sialophosphoprotein(DSPP) were detected by ALP activity assay and Western blotting.Results Compared with DPSC/wt or DPSC/vector separately,proliferating rate and S-cycle of DPSC/Delta1-RNAi was significantly lower.The S phase and proliferation index ( PI ) decreased markedly from 22.32 ± 2.35 and 33.68 ±4.19(DPSC/Delta1-RNAi) to 5.44 ± 0.91 and 16.00 ± 6.07 (DPSC/wt).The PCNA staining of DPSC/Delta1-RNAi was evidently weaker.DPSC/Delta1-RNAi group had more calcified cell nodules than the other two control groups,and ALP activity and DSPP expression of DPSC/Delta1-RNAi group increased markedly.Conclusions Deltal-RNAi induced by the lentivirus vectors may inhibit DPSC proliferation and differentiation.Notch-Delta signal pathway plays an important role in self-renewal and differentiation.

OBJECTIVETo develop an HPLC method for the determination of luteolin-7-O-glycoside, apigenin-7-O-glycoside, linarin, luteolin, apigenin and acacetin in Flos Chrysanthemi Indici simultaneously.

METHODThe reversed phase HPLC system consisting of a C18 column (4.6 mm x 250 mm, 5 microm) and a mixture of acetonitrile and 0.05% phosphate acid with gradient elution as the mobile phase was adopted. The absorbance was monitored at 326 nm.

RESULTThe linear response range was 1.08-21.5 mg x L(-1) (r = 0.9990), 0.278-5.57 mg x L(-1) (r = 0.9994), 5.58-112 mg x L(-1) (r = 0.9999), 0.521-10.4 mg x L(-1) (r = 0.9995), 0.162-3.25 mg x L(-1) (r = 0.9999) and 0.288-5.76 mg x L(-1) (r = 0.9998), respectively (n = 6). The average recoveries (n = 9) of six flavonoids were 96.34%-104.0%. All of RSD of precision (n = 6) and repeatability (n = 6) were less than 1.2% and 2.6%.

CONCLUSIONThe validation data demonstrated that the method was accurate and repeatable, and can be ase to measure the six flavonoids in Flos Chrysanthemi Indici.

Chromatography, High Pressure Liquid ; Chromatography, Reverse-Phase ; Chrysanthemum ; chemistry ; Drugs, Chinese Herbal ; analysis ; Flavonoids ; analysis

Objective To investigate the value of enhanced MR texture analysis in the differential diagnosis of single brain metastasis (SBM) and high grade glioma (HGG). Methods The preoperative MRI images of 21 SBM and 23 HGG patients confirmed by pathology were retrospectively analyzed. The T1WI enhanced images were used for texture analysis to obtain the mean, median, entropy, skewness, kurtosis, heterogeneity and other parameters. An independent sample t-test or Mann-Whitney U test was used to statistically analyze the values of the parameters between the two groups, and ROC evaluation parameters were used to identify the diagnostic efficacy of SBM and HGG. Results There were significant differences in entropy and heterogeneity parameters between SBM and HGG: (4.838 ± 0.523 vs. 5.358 ± 0.449, 0.276 ± 0.066 vs. 0.331 ± 0.101), P＜0.05. When the cut-off value of entropy was 4.88, the sensitivity of differential diagnosis between SBM and HGG was 47.6%, specificity was 100.0%, and the area under ROC curve was the largest (0.745). Conclusions The entropy value and heterogeneity have certain value in distinguishing SBM and HGG in MRI texture analysis.

Objective ToinvestigatethevalueofimagetextureanalysisbasedonconventionalMRimagesinthedifferentialdiagnosisof intracranialsolitaryfibroustumor/hemangiopericytoma(SFT/HPC)andvascularmeningiomas.Methods 12casesofSFT/HPCand 16casesofvascularmeningiomaconfirmedbypathologywerecollected.Variousdiscriminantanalysismethodsandimagetextureanalysis techniqueswereappliedtopre-operativeroutineMRIimagesofthebrain,andthebesttextureparameterswereselectedtoclassify cases,includinglineardiscriminantanalysis(LDA),nonlineardiscriminantanalysis(NDA),principalcomponentanalysis(PCA)and rawdataanalysis(RDA).Finally,thebestclassificationsequencetextureparameterswerechosenforstatisticalanalysis.Results The enhancedT1WIwasthebestclassificationsequence.ThePOE+ACC methodhadthelowestmisclassificationrate.Theabsolutegradient skewness,theautocorrelationofthecooccurrencematrix,andthevarianceofthehistograminthetexturefeaturesoftheSFT/HPC andvascularmeningiomasweredifferent(P<0.05).Conclusion ThatbasedontheconventionalMRimagetextureanalysiscanprovidemore quantitativeinformation,anew methodandideafortheidentificationofintracranialSFT/HPCandvascularmeningioma.

OBJECTIVE: To determine the carrier rate for 21 inherited metabolic diseases among a Chinese population of childbearing age. METHODS: A total of 897 unrelated healthy individuals (including 143 couples) were recruited, and DNA was extracted from their peripheral blood samples. Whole exome sequencing (WES) was carried out to screen potential variants among 54 genes associated with 21 inherited metabolic diseases. Pathogenic and likely pathogenic variants and unreported loss-of-function variants were analyzed. RESULTS: One hundred fourty types of pathogenic/likely pathogenic variants (with an overall number of 183) and unreported loss-of-function variants were detected, which yield a frequency of 0.20 per capita. A husband and wife were both found to carry pathogenic variants of the SLC25A13 gene and have given birth to a healthy baby with the aid of preimplantation genetic diagnosis. The detected variants have involved 40 genes, with the most common ones including ATP7B, SLC25A13, PAH, CBS and MMACHC. Based on the Hardy-Weinberg equilibrium, the incidence of the 21 inherited metabolic diseases in the population was approximately 1/1100, with the five diseases with higher incidence including citrullinemia, methylmalonic acidemia, Wilson disease, glycogen storage disease, and phenylketonuria. CONCLUSION This study has preliminarily determined the carrier rate and incidence of 21 inherited metabolic diseases among a Chinese population of childbearing age, which has provided valuable information for the design of neonatal screening program for inherited metabolic diseases. Pre-conception carrier screening can provide an important measure for the prevention of transmission of Mendelian disorders in the population.
Asians/genetics* ; China ; Exome ; Female ; Humans ; Infant, Newborn ; Metabolic Diseases/genetics* ; Mitochondrial Membrane Transport Proteins/genetics* ; Oxidoreductases/genetics* ; Whole Exome Sequencing

OBJECTIVETo investigate the effects of sleep deprivation on intelligence development in primary school students.

METHODSBetween June 2009 and April 2010, 316 grade 5 students aged 10-11 years were selected from four primary schools in four administrative districts of Changsha, China by stratified random sampling. The intelligence characteristics of children with varying degrees of sleep deprivation were investigated using the Chinese Wechsler Intelligence Scale for Children.

RESULTSA total of 286 valid questionnaires were received, with a response rate of 90.5%. The survey was comprised of a sleep deprivation group (sleep time <8 hours per night; n=180) and a control group (sleep time ≥8 hours per night; n=106). The sleep deprivation group had significantly lower subtest scores, verbal intelligence quotient (IQ) (VIQ), performance IQ (PIQ) and full scale IQ (P<0.05) and significantly lower verbal comprehension factor score and memory/attention factor score compared with the control group (P<0.05). Compared with the control group, the moderate sleep deprivation subgroup had significantly decreased VIQ and full scale IQ as well as verbal comprehension factor score and memory/attention factor score (P<0.05), and the severe sleep deprivation subgroup showed decreases in all scores (P<0.05). The sleep deprivation group and moderate and severe sleep deprivation subgroups had significantly higher proportions of children with VIQ-PIQ imbalance than the control group.

CONCLUSIONSSleep deprivation adversely affects intelligence development, especially VIQ, in primary school students, and the adverse effects of sleep deprivation are mainly seen in students with moderate and severe sleep deprivation.

Child ; Female ; Humans ; Intelligence ; Male ; Sleep Deprivation ; psychology

Aim To investigate the intracellular up-take and target protein binding characteristics of two Bruton's tyrosine kinase inhibitors(BTKi)with differ-ent selectivities to provide further insights into the mechanisms of drug off-target-related bleeding risk.Methods Ibrutinib(non-selective BTKi)and za-nubrutinib(selective BTKi)were used as study drugs.After incubation of MEC-1 cells and human platelets with drugs,the cellular thermal shift assay(CETSA)was combined with Western blot to obtain the melting curve and isothermal curve to analyze the binding char-acteristics of the two drugs with the target protein BTK.After incubation of MEC-1 cells and human platelets with drugs,the concentrations of the two drugs were detected by liquid chromatography-tandem mass spectrometry(LC-MS/MS)to analyze the intracellular uptake of the two drugs.Results CETSA analysis confirmed that zanubrutinib was more selective for the target protein BTK compared to ibrutinib.LC-MS/MS analysis showed that both drugs were uptaken intracel-lularly by MEC-1 cells and platelets in a concentration-dependent manner.Conclusions While BTKi targe-ting BTK to B lymphocytes exerts therapeutic effects,off-target effects on platelets due to differences in their intracellular uptake,and target-binding characteristics may be one of the reasons for the differences in bleed-ing risk across selective BTKi.

OBJECTIVETo investigate the possibility of Hantavirus (HV) and Orientia tsutsugamushi (Ot) coinfection in their hosts.

METHODSHV and Ot were used to infect Vero E6 cells cultured in vitro singly, simultaneously or successively. Genes of HV and Ot were identified in different generation cells with RT-PCR.

RESULTSFive experiment groups of infected Vero E6 cells were tested, the results were as follows: HV and Ot were both positive in infected Vero E6 cells passaged 2 times and the positive rate increased following the passaged times in HV and Ot infection groups, simultaneously or successively. However, in the groups which were infected with HV and Ot separately, the gene of HV or Ot could be detected in infected Vero E6 cells passaged only once and the positive rate increased following the times of the passaged. The positive rate was higher in the singly infected groups than in those infected simultaneously or successively.

CONCLUSIONCoinfection of HV and Ot did exist in the hosts while HV and Ot could inhibit each other in the initial infection stage.

Animals ; Cell Division ; Cercopithecus aethiops ; Hantavirus ; pathogenicity ; Hantavirus Infections ; Orientia tsutsugamushi ; pathogenicity ; Reverse Transcriptase Polymerase Chain Reaction ; Scrub Typhus ; Vero Cells